PMID- 19673806 TI - Chills as an indicator of individual emotional peaks. AB - Chills (goose bumps) have been repeatedly associated with positive emotional peaks. Chills seem to be related to distinct musical structures and the reward system in the brain. A new approach that uses chills as indicators of individual emotional peaks is discussed. Chill reactions of 95 participants in response to seven music pieces were recorded. Subjective intensity as well as physiological arousal (skin conductance response, heart rate) revealed peaks during chill episodes. This review suggests that chills are a reliable indicator of individual emotional peaks, combining reports of subjective feelings with physiological arousal. PMID- 19673807 TI - Electrophysiological correlates of aesthetic music processing: comparing experts with laypersons. AB - We analyzed the processes of making aesthetic judgments of music, focusing on the differences between music experts and laypersons. Sixteen students of musicology and 16 control subjects (also students) judged the aesthetic value as well as the harmonic correctness of chord sequences. Event-related potential (ERP) data indicate differences between experts and laypersons in making aesthetic judgments at three different processing stages. Additionally, effects of expertise on ERP components that have previously been proven to be sensitive to musical training were replicated. The study thus provides insights into the effects of musical expertise on neural correlates of aesthetic music processing. PMID- 19673808 TI - Emotion-related changes in heart rate and its variability during performance and perception of music. AB - The present study investigated the differential effects of emotions evoked by music on heart rate (HR) and its variability (HRV) during the playing of music on the piano compared to those in persons listening to the same music. Thirteen elite pianists underwent experiments under expressive piano playing, nonexpressive piano playing, expressive listening, and nonexpressive listening conditions. The expressive conditions produced significantly higher levels of HR and low-frequency component of HRV, as well as a lower level of its high frequency component. A greater modulation of these was also revealed for performance than perception. The findings suggested that musical performance would lead to a greater effect of emotion-related modulation in cardiac autonomic nerve activity than musical perception. PMID- 19673809 TI - The influence of social situations on music listening. AB - The aim of this study was to investigate whether listening to music in a group setting influenced the emotion felt by the listeners. We hypothesized that individuals hearing music in a group would experience more intense emotions than the same individuals hearing the same music on their own. The emotional reactions to 10 musical excerpts (previously shown to contain chill-inducing psychoacoustic parameters) were measured in a within-subjects design. We found, contrary to our hypothesis, that the participants (all musicians) did not experience more chills when listening to music in a group than when listening alone. These findings may be explained by a lesser degree of concentration on the music in the group condition. PMID- 19673810 TI - Motor/autonomic stress responses in a competitive piano performance. AB - The present study examined the effects of psychological stress on performance quality, autonomic responses, and upper extremity muscle activity in skilled pianists through comparisons between stressful (competition) and nonstressful (rehearsal) conditions. We observed increased levels of subjective anxiety, autonomic arousal, and electromyographic activity in the competition condition, which could contribute to the impairment of performance quality. The results provide important practical implications for enhancing performance quality as well as preventing playing-related musculoskeletal disorders in musicians. PMID- 19673811 TI - Part VI introduction: listening to and making music facilitates brain recovery processes. PMID- 19673812 TI - A neuroscientific perspective on music therapy. AB - During the last years, a number of studies demonstrated that music listening (and even more so music production) activates a multitude of brain structures involved in cognitive, sensorimotor, and emotional processing. For example, music engages sensory processes, attention, memory-related processes, perception-action mediation ("mirror neuron system" activity), multisensory integration, activity changes in core areas of emotional processing, processing of musical syntax and musical meaning, and social cognition. It is likely that the engagement of these processes by music can have beneficial effects on the psychological and physiological health of individuals, although the mechanisms underlying such effects are currently not well understood. This article gives a brief overview of factors contributing to the effects of music-therapeutic work. Then, neuroscientific studies using music to investigate emotion, perception-action mediation ("mirror function"), and social cognition are reviewed, including illustrations of the relevance of these domains for music therapy. PMID- 19673813 TI - Evidence for plasticity in white-matter tracts of patients with chronic Broca's aphasia undergoing intense intonation-based speech therapy. AB - Recovery from aphasia can be achieved through recruitment of either perilesional brain regions in the affected hemisphere or homologous language regions in the nonlesional hemisphere. For patients with large left-hemisphere lesions, recovery through the right hemisphere may be the only possible path. The right-hemisphere regions most likely to play a role in this recovery process are the superior temporal lobe (important for auditory feedback control), premotor regions/posterior inferior frontal gyrus (important for planning and sequencing of motor actions and for auditory-motor mapping), and the primary motor cortex (important for execution of vocal motor actions). These regions are connected reciprocally via a major fiber tract called the arcuate fasciculus (AF), however, this tract is not as well developed in the right hemisphere as it is in the dominant left. We tested whether an intonation-based speech therapy (i.e., melodic intonation therapy [MIT]), which is typically administered in an intense fashion with 75-80 daily therapy sessions, would lead to changes in white-matter tracts, particularly the AF. Using diffusion tensor imaging (DTI), we found a significant increase in the number of AF fibers and AF volume comparing post- with pretreatment assessments in six patients that could not be attributed to scan-to-scan variability. This suggests that intense, long-term MIT leads to remodeling of the right AF and may provide an explanation for the sustained therapy effects that were seen in these six patients. PMID- 19673814 TI - Neural reorganization underlies improvement in stroke-induced motor dysfunction by music-supported therapy. AB - Motor impairments are common after stroke, but efficacious therapies for these dysfunctions are scarce. By extending an earlier study on the effects of music supported therapy, behavioral indices of motor function as well as electrophysiological measures were obtained before and after a series of therapy sessions to assess whether this new treatment leads to neural reorganization and motor recovery in patients after stroke. The study group comprised 32 stroke patients in a large rehabilitation hospital; they had moderately impaired motor function and no previous musical experience. Over a period of 3 weeks, these patients received 15 sessions of music-supported therapy using a manualized step by-step approach. For comparison 30 additional patients received standard rehabilitation procedures. Fine as well as gross motor skills were trained by using either a MIDI-piano or electronic drum pads programmed to emit piano tones. Motor functions were assessed by an extensive test battery. In addition, we studied event-related desynchronization/synchronization and coherences from all 62 patients performing self-paced movements of the index finger (MIDI-piano) and of the whole arm (drum pads). Results showed that music-supported therapy yielded significant improvement in fine as well as gross motor skills with respect to speed, precision, and smoothness of movements. Neurophysiological data showed a more pronounced event-related desynchronization before movement onset and a more pronounced coherence in the music-supported therapy group in the post-training assessment, whereas almost no differences were observed in the control group. Thus we see that music-supported therapy leads to marked improvements of motor function after stroke and that these are accompanied by electrophysiological changes indicative of a better cortical connectivity and improved activation of the motor cortex. PMID- 19673815 TI - Neurologic music therapy improves executive function and emotional adjustment in traumatic brain injury rehabilitation. AB - This study examined the immediate effects of neurologic music therapy (NMT) on cognitive functioning and emotional adjustment with brain-injured persons. Four treatment sessions were held, during which participants were given a pre-test, participated in 30 min of NMT that focused on one aspect of rehabilitation (attention, memory, executive function, or emotional adjustment), which was followed by post-testing. Control participants engaged in a pre-test, 30 min of rest, and then a post-test. Treatment participants showed improvement in executive function and overall emotional adjustment, and lessening of depression, sensation seeking, and anxiety. Control participants improved in emotional adjustment and lessening of hostility, but showed decreases in measures of memory, positive affect, and sensation seeking. PMID- 19673816 TI - A new approach to rhythm cueing of cognitive functions: the case of ideomotor apraxia. AB - Although positive effects of rhythm cueing on motor control in neurologic disorders are known, no studies have yet focused on patients suffering from impaired programming of complex actions. One patient suffering from ideomotor apraxia (a potentially ideal experimental paradigm to test the effect of rhythm on high-level motor control) underwent two rehabilitation training sets differing only for the presence or absence of rhythm cueing. Both sets of training increased the patient's proficiency, but rhythm cueing was significantly more effective, during the training as well as during the post-training uncued test. Ideomotor apraxia represents an effective model to test the effects of rhythm on high-level motor control. PMID- 19673817 TI - Music programs designed to remedy burnout symptoms show significant effects after five weeks. AB - Earlier studies have demonstrated that music interventions can lessen symptoms of depression. Depression and burnout are closely related. We hypothesized that specially designed receptive music therapy programs and protocols might reduce the symptoms of burnout. In a four-arm randomized, placebo- and waiting-list controlled double-blind study, including 150 participants, two specific music programs significantly reduced burnout symptoms after 5 weeks. The effects were maintained over a long time period. This newly developed method of receptive music therapy was also evaluated for the treatment of depression and dysthymia, with significant outcomes. PMID- 19673818 TI - Therapeutic role of music listening in stroke rehabilitation. AB - We performed two parallel interview studies of stroke patients (n= 20) and professional nurses (n= 5) to gain more insight into the therapeutic role of music listening in stroke rehabilitation. Results suggest that music listening can be used to relax, improve mood, and provide both physical and mental activation during the early stages of recovery from stroke. Thus, music listening could provide a useful clinical tool in stroke rehabilitation. PMID- 19673819 TI - Melodic intonation therapy: shared insights on how it is done and why it might help. AB - For more than 100 years, clinicians have noted that patients with nonfluent aphasia are capable of singing words that they cannot speak. Thus, the use of melody and rhythm has long been recommended for improving aphasic patients' fluency, but it was not until 1973 that a music-based treatment [Melodic Intonation Therapy (MIT)] was developed. Our ongoing investigation of MIT's efficacy has provided valuable insight into this therapy's effect on language recovery. Here we share those observations, our additions to the protocol that aim to enhance MIT's benefit, and the rationale that supports them. PMID- 19673820 TI - Reestablishing speech understanding through musical ear training after cochlear implantation: a study of the potential cortical plasticity in the brain. AB - Cochlear implants (CIs) provide impressive speech perception for persons with severe hearing loss, but many CI recipients fail in perceiving speech prosody and music. Successful rehabilitation depends on cortical plasticity in the brain and postoperative measures. The present study evaluates the behavioral and neurologic effects of musical ear training on CI users' speech and music perception. The goal is to find and work out musical methods to improve CI users' auditory capabilities and, in a longer perspective, provide an efficient strategy for improving speech understanding for both adults and children with CIs. PMID- 19673821 TI - Amusia and cognitive deficits after stroke: is there a relationship? AB - We studied the relationship between musical and cognitive deficits by testing middle cerebral arterial (MCA) stroke patients (n= 53) with a shortened version of the Montreal Battery of Evaluation of Amusia (MBEA) and an extensive neuropsychological test battery. Results showed that amusic patients (n= 32) had more severe cognitive deficits, especially in working memory and executive functioning, than did non-amusic patients (n= 21), and the severity of amusia also correlated with attention deficits. These findings thus suggest that domain general attention, executive, and working memory processes are associated with amusia after stroke. PMID- 19673822 TI - Part VII introduction: music, language, and motor programming: a common neural organization? PMID- 19673823 TI - Broca's area in language, action, and music. AB - The work of Paul Broca has been of pivotal importance in the localization of some higher cognitive brain functions. He first reported that lesions to the caudal part of the inferior frontal gyrus were associated with expressive deficits. Although most of his claims are still true today, the emergence of novel techniques as well as the use of comparative analyses prompts modern research for a revision of the role played by Broca's area. Here we review current research showing that the inferior frontal gyrus and the ventral premotor cortex are activated for tasks other than language production. Specifically, a growing number of studies report the involvement of these two regions in language comprehension, action execution and observation, and music execution and listening. Recently, the critical involvement of the same areas in representing abstract hierarchical structures has also been demonstrated. Indeed, language, action, and music share a common syntactic-like structure. We propose that these areas are tuned to detect and represent complex hierarchical dependencies, regardless of modality and use. We speculate that this capacity evolved from motor and premotor functions associated with action execution and understanding, such as those characterizing the mirror-neuron system. PMID- 19673824 TI - Studying synchronization to a musical beat in nonhuman animals. AB - The recent discovery of spontaneous synchronization to music in a nonhuman animal (the sulphur-crested cockatoo Cacatua galerita eleonora) raises several questions. How does this behavior differ from nonmusical synchronization abilities in other species, such as synchronized frog calls or firefly flashes? What significance does the behavior have for debates over the evolution of human music? What kinds of animals can synchronize to musical rhythms, and what are the key methodological issues for research in this area? This paper addresses these questions and proposes some refinements to the "vocal learning and rhythmic synchronization hypothesis." PMID- 19673825 TI - Effects of context on electrophysiological response to musical accents. AB - Listeners' aesthetic and emotional responses to music typically occur in the context of long musical passages that contain structures defined in terms of the events that precede them. We describe an electrophysiological study of listeners' brain responses to musical accents that coincided in longer musical sequences. Musically trained listeners performed a timbre-change detection task in which a single-tone timbre change was positioned within 4-bar melodies composed of 350-ms tones to coincide or not with melodic contour accents and temporal accents (induced with temporal gaps). Event-related potential responses to (task relevant) attended timbre changes elicited an early negativity (MMN/N2b) around 200 ms and a late positive component around 350 ms (P300), reflecting updating of the timbre change in working memory. The amplitudes of both components changed systematically across the sequence, consistent with expectancy-based context effects. Furthermore, melodic contour changes modulated the MMN/N2b response (but not the P300) to timbre changes in later sequence positions. In contrast, task irrelevant temporal gaps elicited an MMN that was not modulated by position within the context; absence of a P300 indicated that temporal-gap accents were not updated in working memory. Listeners' neural responses to musical structure changed systematically as sequential predictability and listeners' expectations changed across the melodic context. PMID- 19673826 TI - Integrated preattentive processing of vowel and pitch: a mismatch negativity study. AB - This study examines the additivity of the Mismatch Negativity (MMN) as an index of the early interactions between vowels and pitch when sung. Event-related potentials (ERPs) were recorded while participants were presented with sung vowels. Sixteen percent of stimuli deviated in pitch only, in vowel only, or in both pitch and vowel. All three kinds of deviants elicited an MMN of similar amplitude. The MMNs to vowel and pitch deviants did not show significant additivity. This suggests that vowel and pitch are processed by shared neural substrates at the preattentive level. PMID- 19673827 TI - Similarity and repetition: an ERP study on musical form perception. AB - In this ERP study we investigate how consecutive phrase patterns in the musical form types AABB and ABAB are conjoined perceptually. An anterior N300 reflects recognition of motif similarity in adjacent (A-A) and nonadjacent (A-B-A) phrase patterns, indicating perceptual coherence between (similar) parts in linear order. A marginally significant late anterior negativity reflects the processing of the second B in ABAB. It probably reveals some global aspects of "structural integration" when nonadjacent parts are perceived in the context of the whole melody. PMID- 19673828 TI - Tonal language processing in congenital amusia. AB - Twenty amusic and 20 control speakers of French were presented with pairs of Mandarin lexical tones to discriminate as same or different. Results revealed that even if the amusic group performed significantly below the control group, the scores of the two groups largely overlapped, with only 15% of the amusic group performing outside the normal variations. Thus, the findings suggest a modest transfer of deficit between music and speech, which in turn calls for further work in order to identify the nature of the mediating factors. PMID- 19673829 TI - Overlap of musical and linguistic syntax processing: intracranial ERP evidence. AB - The present study investigated the co-localization of musical and linguistic syntax processing in the human brain. EEGs were recorded from subdural electrodes placed on the left and right perisylvian cortex. The neural generators of the early potentials elicited by syntactic errors in music and language were localized by means of distributed source modeling and compared within subjects. The combined results indicated a partial overlap of the sources within the bilateral superior temporal gyrus, and, to a lesser extent, in the left inferior frontal gyrus, qualifying these areas as shared anatomic substrates of early syntactic error detection in music and language. PMID- 19673830 TI - Self-recognition in the perception of actions performed in synchrony with music. AB - This study investigated self-recognition in point-light displays depicting actions performed in synchrony with music. Participants were recorded executing three different actions (dancing, walking, and clapping) and were subsequently required to identify the agent (self versus other) from point-light displays with or without the accompanying music. Results indicate that while recognition accuracy was better than chance for all actions, it was best for the relatively complex dance actions. The presence of music did not affect accuracy, suggesting that self-recognition was based on information about personal movement kinematics rather than individual differences in synchrony between movements and music. PMID- 19673831 TI - Spatial properties of perceived pitch: influence on reaching movements. AB - We present three experiments that show a clear influence of pitch on reaching movements. The first experiment shows that when there are incompatibilities between the spatial representations of target coordinates and perceived pitches, response times are longer than when spatial representations are congruous. The second experiment shows that pitch can influence motor responses, biasing their direction. The third experiment shows that in the absence of any other spatial cues, tones are assigned spatial coordinates based on frequency. Our results suggest that pitch may indeed have inherent spatial properties, and that these properties may affect movement planning and execution. PMID- 19673832 TI - Infants detect cross-modal cues to identity in speech and singing. AB - Little is known about infants' perception of cross-modal cues to identity, but the importance of recognizing familiar individuals makes it likely that this skill would be evident early in life. Infants 6-8 months of age were tested on their ability to link dynamic cross-modal cues to the identity of unfamiliar speakers and singers. After exposure to speech or singing, infants watched two silent videos, one featuring the previously heard speaker or singer. Infants looked significantly longer at the video of the person heard previously, which indicates that they can match auditory and visual cues to the identity of unfamiliar persons. PMID- 19673833 TI - Motor timing and precursor literacy skills in very young children. AB - It is generally agreed that efficiency in processing basic speech sounds is a strong indicator of literacy outcomes. However, there is some debate about the extent to which this capacity is encapsulated within the language system. We report results based on pretest data obtained in an early music training study investigating the relationship between a motor-rhythmic measure of ability (synchronous tapping) and the purported purely linguistically based processing measures, Rapid Automatized Naming (RAN) and Phonological Awareness (PA). Results suggest that RAN ability relies to some extent on nonlinguistic temporal processing skills. PMID- 19673834 TI - Part VIII introduction: new directions: cochlear implants. PMID- 19673835 TI - Melodic contour identification and music perception by cochlear implant users. AB - Research and outcomes with cochlear implants (CIs) have revealed a dichotomy in the cues necessary for speech and music recognition. CI devices typically transmit 16-22 spectral channels, each modulated slowly in time. This coarse representation provides enough information to support speech understanding in quiet and rhythmic perception in music, but not enough to support speech understanding in noise or melody recognition. Melody recognition requires some capacity for complex pitch perception, which in turn depends strongly on access to spectral fine structure cues. Thus, temporal envelope cues are adequate for speech perception under optimal listening conditions, while spectral fine structure cues are needed for music perception. In this paper, we present recent experiments that directly measure CI users' melodic pitch perception using a melodic contour identification (MCI) task. While normal-hearing (NH) listeners' performance was consistently high across experiments, MCI performance was highly variable across CI users. CI users' MCI performance was significantly affected by instrument timbre, as well as by the presence of a competing instrument. In general, CI users had great difficulty extracting melodic pitch from complex stimuli. However, musically experienced CI users often performed as well as NH listeners, and MCI training in less-experienced subjects greatly improved performance. With fixed constraints on spectral resolution, such as occurs with hearing loss or an auditory prosthesis, training and experience can provide considerable improvements in music perception and appreciation. PMID- 19673836 TI - Music in the lives of deaf children with cochlear implants. AB - Present-day cochlear implants provide good temporal cues and coarse spectral cues. In general, these cues are adequate for perceiving speech in quiet backgrounds and for young children's acquisition of spoken language. They are inadequate, however, for conveying the rich pitch-patterning of music. As a result, many adults who become implant users after losing their hearing find music disappointing or unacceptable. By contrast, child implant users who were born deaf or became deaf as infants or toddlers typically find music interesting and enjoyable. They recognize popular songs that they hear regularly when the test materials match critical features of the original versions. For example, they can identify familiar songs from the original recordings with words and from versions that omit the words but preserve all other cues. They also recognize theme songs from their favorite television programs when presented in original or somewhat altered form. The motivation of children with implants for listening to music or melodious speech is evident well before they understand language. Within months after receiving their implant, they prefer singing to silence. They also prefer speech in the maternal style to typical adult speech and the sounds of their native language-to-be to those of a foreign language. An important task of future research is to ascertain the relative contributions of perceptual and motivational factors to the apparent differences between child and adult implant users. PMID- 19673838 TI - Neuromusic III. Proceedings of the Third Neuroscience and Music Conference. June 2009. Montreal, Canada. PMID- 19673840 TI - In this issue--August 2009. PMID- 19673837 TI - Experience-induced malleability in neural encoding of pitch, timbre, and timing. AB - Speech and music are highly complex signals that have many shared acoustic features. Pitch, Timbre, and Timing can be used as overarching perceptual categories for describing these shared properties. The acoustic cues contributing to these percepts also have distinct subcortical representations which can be selectively enhanced or degraded in different populations. Musically trained subjects are found to have enhanced subcortical representations of pitch, timbre, and timing. The effects of musical experience on subcortical auditory processing are pervasive and extend beyond music to the domains of language and emotion. The sensory malleability of the neural encoding of pitch, timbre, and timing can be affected by lifelong experience and short-term training. This conceptual framework and supporting data can be applied to consider sensory learning of speech and music through a hearing aid or cochlear implant. PMID- 19673841 TI - Re: Improved traceability of pigs in Australia. PMID- 19673843 TI - Re: Trick or treatment. PMID- 19673844 TI - Risk factors for post-weaning mortality of Merino sheep in south-eastern Australia. AB - OBJECTIVE: To measure associations between body weight, growth rate, sex, time of shearing and post-weaning mortality of Merino sheep. DESIGN: Uni- and multivariable survival analyses of sheep mortality during the first year after weaning, using records (n = 3657) from two field experiments conducted in Western Victoria from 1996 to 2003. RESULTS: Overall mortality was 14.3% (range 4.5 26.8%) and mean maximum mortality rate was 29 deaths/1000 weaners/month. Increased mortality risk was associated with decreases in fleece-free body weight and mean weaner growth rate, particularly at low weights and growth rates. Weaners in the lightest weaning weight quintile had a hazard ratio of 3.5, compared with the middle quintile. The hazard ratio for a 2-kg decrease in weaning weight was 1.2 to 1.7 for weaners lighter than 22 kg. The hazard ratio for a reduction in mean weaner growth rate in the first 5 months after weaning of 0.25 kg/month was 1.1 to 6.8 if mean growth rate was less than 1 kg/month, but did not differ significantly from 1 at greater growth rates. The hazard ratio for wether weaners was approximately 1.5 compared with ewe weaners. The hazard ratio for weaners shorn between December and May, compared with unshorn weaners, was 1.2 to 3.5, with the greatest risk difference associated with shearing in March (45 deaths/1000 weaners/month). CONCLUSION: Improving the body weight and mean growth rate of weaner sheep is likely to reduce post-weaning mortality. Lightweight weaners in a flock should be managed separately from the main portion after weaning. In southern Australia, not shearing spring-born Merino weaners between December and May may assist in reducing overall post-weaning mortality. PMID- 19673845 TI - Transcutaneous ultrasound over the right flank to diagnose mid- to late-pregnancy in the dairy cow. AB - OBJECTIVE: To investigate the practical application and image quality of transcutaneous ultrasonography across the right flank for the diagnosis of mid- to late-gestation in cattle. PROCEDURE: Dairy cows of known pregnancy status were used to establish criteria for the identification of mid- to late-stage pregnancy by transcutaneous ultrasonography. Factors involved in the practical application of transcutaneous ultrasound for routine use in pregnancy diagnosis in dairy cattle were also examined. RESULTS: Transcutaneous ultrasound, using a 3.5-MHz sector transducer applied over the caudodorsal right flank fold, allowed rapid and detailed visualisation of bovine pregnancy (i.e. fetus, fetal fluids, placentomes and/or fetal membranes) and could be used to establish criteria for the diagnosis of pregnancy. The commonly available cattle holding facilities of the herringbone dairy, rotary herringbone dairy, AI race or crush, and walk through dairy all allow safe access to the right flank of a dairy or beef cow. However, transcutaneous scanning over the right flank is difficult in modern rotary turnstile dairies. CONCLUSION: A 3.5-MHz sector transducer applied over the caudodorsal right flank fold allows detailed visualisation of the bovine fetus in most types of cattle-holding facilities. Further investigation of the sensitivity and specificity of this technique over the entire gestation period and its possible use in ageing of the bovine fetus is merited. PMID- 19673847 TI - Comparison of transcutaneous ultrasound over the right flank with transrectal ultrasound for pregnancy diagnosis in the dairy cow. AB - OBJECTIVE: Evaluate the sensitivity and specificity of transrectal ultrasound and transcutaneous ultrasonography across the right flank between days 31 and 196 of pregnancy in the dairy cow. PROCEDURE: Pregnancy status and stage of gestation at date of pregnancy diagnosis by transcutaneous and transrectal ultrasonography were determined for 1570 dairy cattle. Sensitivity and specificity values and probabilities of a correct diagnosis for both techniques were determined and compared. Possible effects of gestational age, herd, method of transcutaneous ultrasound diagnosis, cow age, cow and bull breed, and calf sex on the determination of a correct diagnosis of pregnancy status were evaluated for both techniques. RESULTS: The overall sensitivity and probability of a correct diagnosis of pregnancy status from days 31 to 196 of gestation were significantly higher when using transrectal ultrasound relative to transcutaneous ultrasound over the right flank. However, the sensitivity of transcutaneous ultrasound changed over the course of pregnancy: it was low (7-8%) before day 84, increased to 89% between days 141 and 154, and was close to 100% thereafter. The specificity of both methods did not differ significantly. Herd and gestational age at date of pregnancy diagnosis were significantly associated with making a correct test result using transcutaneous ultrasound, although only the latter predictor was significantly associated with making a correct test result using transrectal ultrasound. CONCLUSION: Transcutaneous ultrasound can not be recommended as an accurate method for early pregnancy diagnosis in the dairy cow, particularly on a whole-herd basis. However, if pregnancy testing is undertaken in mid to late gestation, this technique could provide an accurate and rapid alternative to transrectal ultrasound or manual palpation per rectum. PMID- 19673848 TI - Ultrasonographic characteristics of soft tissue tumours in dogs. AB - OBJECTIVE: To identify and describe the ultrasonographic features of soft tissue tumours in dogs. PROCEDURE: Superficial soft tissue tumours of various histological types, including mast cell tumours (MCTs) and soft tissue sarcomas (STSs), were evaluated. Ultrasound was used to visualise internal characteristics of the tumour, including vascularity. Tumours were categorised according to size, shape, margin definition, tissue plane mobility, echogenicity, echotexture, acoustic shadowing or enhancement and vessel distribution. Objective measurements of intratumoural blood flow included velocities and maximal perfused cross sectional area (fractional area). Logistic regression models incorporating a variety of data were used in an attempt to predict the histopathological type of tumours. RESULTS: The logistic regression model defined by the parameters echotexture, margin definition and presence of subcapsular vessels was highly predictive of MCTs (> 73%; P = 0.024). Several other trends, including a larger size for STSs and less vascularity for both MCTs and STSs, were observed, but did not reach statistical significance. CONCLUSION: This preliminary study has shown the potential diagnostic value of ultrasound in differentiating soft tissue tumours. However, at present, ultrasound cannot replace biopsy and histopathological evaluation for tumour diagnosis. PMID- 19673849 TI - Clinical remission of idiopathic hypereosinophilic syndrome in a Rottweiler. AB - Idiopathic hypereosinophilic syndrome (HES) is a rare syndrome for which Rottweilers appear to over-represent the canine cases. A 6-month-old female entire Rottweiler presented with seizures following a traumatic incident. The dog was identified as having a marked, sustained eosinophilia and investigations did not identify an underlying cause. Concurrently, the dog had chronic eosinophilic hepatitis with impaired liver function and mesenteric eosinophilic lymphadenitis. The dog went on to have spontaneous resolution of HES and normal liver function was subsequently documented. To date, the dog is still alive, more than 4 years after initial presentation. The diagnosis of idiopathic HES in Rottweilers may not carry a poor prognosis and the condition may spontaneously resolve, even in cases with documented organ damage. PMID- 19673850 TI - Toxicity in three dogs from accidental oral administration of a topical endectocide containing moxidectin and imidacloprid. AB - Three dogs were presented with a history of oral administration of a topical endectocide containing imidacloprid and moxidectin. They were diagnosed with imidacloprid and moxidectin intoxication, having ingested doses ranging from 7.5 to 1.4 mg/kg of imidacloprid and 1.9 to 2.8 mg/kg of moxidectin. The three dogs were affected to different degrees of severity, but all displayed signs of ataxia, generalised muscle tremors, paresis, hypersalivation and disorientation. Temporary blindness occurred in two cases. The three dogs were tested for the presence of the multi-drug resistance 1 gene deletion, which can cause an increased sensitivity to the toxic effects of moxidectin, and were found to be negative. Treatment included gastrointestinal decontamination, intravenous fluid therapy and benzodiazepines to control muscle tremors. All three dogs made a complete recovery within 48 h of ingestion. PMID- 19673851 TI - Scrub-itch mite infestation in the endangered bridled nailtail wallaby. AB - Skin lesions on the ears and inguinal and axillary regions of a number of adult animals within a captive population of the endangered bridled nailtail wallaby (Onychogalea fraenata) were associated with the trombiculid mite, Eutrombicula hirsti. The local inflammatory response of these Australian marsupials is described. PMID- 19673854 TI - Bioinformatics and ethics. PMID- 19673852 TI - Late onset cerebellar cortical degeneration in a koala. AB - A 10-year-old male koala started to fall from the tree while sleeping. Subsequently, the koala often fell down while walking and showed a gait abnormality, abnormal nystagmus and hypersalivation. At 12 years of age, the koala became ataxic and seemed blind. At 13 years of age, the koala exhibited signs of dysstasia and was euthanased. Necropsy revealed marked symmetrical atrophy of the cerebellum. Histopathologically, a severe loss of Purkinje and granule cells was evident in the cerebellum, while the molecular layer was more cellular than normal with cells resembling small neurons, which were positively stained with parvalbumin immunohistochemistry. Reactive Bergmann glial cells (astrocytes) were present adjacent to the depleted Purkinje cell zone. The very late onset and slow progression of the cerebellar cortical degeneration in this case is particularly interesting and appears to be the first report in the koala. PMID- 19673855 TI - Outlining ethical issues in nanotechnologies. AB - Nanotechnologies are an expression of the human ability to control and manipulate matter on a very small scale. Their use will enable an even and constant monitoring of human organisms, in a new and perhaps less invasive way. Debates at all levels--national, European and international--have pointed out the common difficulty of giving a complete, clear definition of nanotechnologies. This is primarily due to the variety of their components, to the fact that there is not just one technology but several. The most significant medical applications of nanotechnologies are in the diagnostic and the therapeutic fields, eg biosensors and molecular imaging, providing diagnosis and drug delivery with no invasive methods involved. Like any other emerging field, such technologies imply new possibilities for improving health but, on the other hand, they are still at an experimental stage and therefore should be implemented under rigorous safety testing before going on general release. For this purpose, the ethical, legal and social implications (ELSI) of nanotechnologies have been elaborated by study groups, in order to develop solutions before the results of the tests are diffused into medical practice. The aim of this paper is to define some of the ethical issues concerning biomedical applications and to evaluate whether there is a need for new or additional guidelines and regulations. PMID- 19673856 TI - A meta-analysis of parasite virulence in nestling birds. AB - Parasitism is a common cause of host mortality, but little is known about the ecological factors affecting parasite virulence (the rate of mortality among infected hosts). We reviewed 117 field estimates of parasite-induced nestling mortality in birds, showing that there was significant consistency in mortality among host and parasite taxa. Virulence increased towards the tropics in analyses of both species-specific data and phylogenetic analyses. We found evidence of greater parasite prevalence being associated with reduced virulence. Furthermore, bird species breeding in open nest sites suffered from greater parasite-induced mortality than hole-nesting species. By contrast, parasite specialization and generation time of parasites relative to that of hosts explained little variation in virulence. Likewise, there were little or no significant effects of host genetic variability, host sociality, host migration, host insular distribution or host survival on parasite virulence. These findings suggest that parasite-induced nestling mortality in birds is mainly determined by geographical location and to a smaller extent nest site and prevalence. PMID- 19673857 TI - The integration of digestion and osmoregulation in the avian gut. AB - We review digestion and osmoregulation in the avian gut, with an emphasis on the ways these different functions might interact to support or constrain each other and the ways they support the functioning of the whole animal in its natural environment. Differences between birds and other vertebrates are highlighted because these differences may make birds excellent models for study and may suggest interesting directions for future research. At a given body size birds, compared with mammals, tend to eat more food but have less small intestine and retain food in their gastrointestinal tract (GIT) for shorter periods of time, despite generally higher mass-specific energy demands. On most foods, however, they are not less efficient at digestion, which begs the question how they compensate. Intestinal tissue-specific rates of enzymatic breakdown of substrates and rates of active transport do not appear higher in birds than in mammals, nor is there a demonstrated difference in the extent to which those rates can be modulated during acclimation to different feeding regimes (e.g. diet, relative intake level). One compensation appears to be more extensive reliance on passive nutrient absorption by the paracellular pathway, because the avian species studied so far exceed the mammalian species by a factor of at least two- to threefold in this regard. Undigested residues reach the hindgut, but there is little evidence that most wild birds recover microbial metabolites of nutritional significance (essential amino acids and vitamins) by re-ingestion of faeces, in contrast to many hindgut fermenting mammals and possibly poultry. In birds, there is some evidence for hindgut capacity to breakdown either microbial protein or protein that escapes the small intestine intact, freeing up essential amino acids, and there is considerable evidence for an amino acid absorptive capacity in the hindgut of both avian and mammalian hindgut fermenters. Birds, unlike mammals, do not excrete hyperosmotic urine (i.e. more than five times plasma osmotic concentration). Urine is mixed with digesta rather than directly eliminated, and so the avian gut plays a relatively more important role in water and salt regulation than in mammals. Responses to dehydration and high- and low salt loads are reviewed. Intestinal absorption of ingested water is modulated to help achieve water balance in one species studied (a nectar-feeding sunbird), the first demonstration of this in any terrestrial vertebrate. In many wild avian species the size and digestive capacity of the GIT is increased or decreased by as much as 50% in response to nutritional challenges such as hyperphagia, food restriction or fasting. The coincident impacts of these changes on osmoregulatory or immune function of the gut are poorly understood. PMID- 19673858 TI - Semiparametric bayesian analysis of nutritional epidemiology data in the presence of measurement error. AB - We propose a semiparametric Bayesian method for handling measurement error in nutritional epidemiological data. Our goal is to estimate nonparametrically the form of association between a disease and exposure variable while the true values of the exposure are never observed. Motivated by nutritional epidemiological data, we consider the setting where a surrogate covariate is recorded in the primary data, and a calibration data set contains information on the surrogate variable and repeated measurements of an unbiased instrumental variable of the true exposure. We develop a flexible Bayesian method where not only is the relationship between the disease and exposure variable treated semiparametrically, but also the relationship between the surrogate and the true exposure is modeled semiparametrically. The two nonparametric functions are modeled simultaneously via B-splines. In addition, we model the distribution of the exposure variable as a Dirichlet process mixture of normal distributions, thus making its modeling essentially nonparametric and placing this work into the context of functional measurement error modeling. We apply our method to the NIH AARP Diet and Health Study and examine its performance in a simulation study. PMID- 19673859 TI - Joint inference on HIV viral dynamics and immune suppression in presence of measurement errors. AB - In an attempt to provide a tool to assess antiretroviral therapy and to monitor disease progression, this article studies association of human immunodeficiency virus (HIV) viral suppression and immune restoration. The data from a recent acquired immune deficiency syndrome (AIDS) study are used for illustration. We jointly model HIV viral dynamics and time to decrease in CD4/CD8 ratio in the presence of CD4 process with measurement errors, and estimate the model parameters simultaneously via a method based on a Laplace approximation and the commonly used Monte Carlo EM algorithm. The approaches and many of the points presented apply generally. PMID- 19673860 TI - Incorporating correlation for multivariate failure time data when cluster size is large. AB - We propose a new estimation method for multivariate failure time data using the quadratic inference function (QIF) approach. The proposed method efficiently incorporates within-cluster correlations. Therefore, it is more efficient than those that ignore within-cluster correlation. Furthermore, the proposed method is easy to implement. Unlike the weighted estimating equations in Cai and Prentice (1995, Biometrika 82, 151-164), it is not necessary to explicitly estimate the correlation parameters. This simplification is particularly useful in analyzing data with large cluster size where it is difficult to estimate intracluster correlation. Under certain regularity conditions, we show the consistency and asymptotic normality of the proposed QIF estimators. A chi-squared test is also developed for hypothesis testing. We conduct extensive Monte Carlo simulation studies to assess the finite sample performance of the proposed methods. We also illustrate the proposed methods by analyzing primary biliary cirrhosis (PBC) data. PMID- 19673861 TI - Longitudinal studies of binary response data following case-control and stratified case-control sampling: design and analysis. AB - We discuss design and analysis of longitudinal studies after case-control sampling, wherein interest is in the relationship between a longitudinal binary response that is related to the sampling (case-control) variable, and a set of covariates. We propose a semiparametric modeling framework based on a marginal longitudinal binary response model and an ancillary model for subjects' case control status. In this approach, the analyst must posit the population prevalence of being a case, which is then used to compute an offset term in the ancillary model. Parameter estimates from this model are used to compute offsets for the longitudinal response model. Examining the impact of population prevalence and ancillary model misspecification, we show that time-invariant covariate parameter estimates, other than the intercept, are reasonably robust, but intercept and time-varying covariate parameter estimates can be sensitive to such misspecification. We study design and analysis issues impacting study efficiency, namely: choice of sampling variable and the strength of its relationship to the response, sample stratification, choice of working covariance weighting, and degree of flexibility of the ancillary model. The research is motivated by a longitudinal study following case-control sampling of the time course of attention deficit hyperactivity disorder (ADHD) symptoms. PMID- 19673862 TI - Joint Spatial Modeling of Recurrent Infection and Growth with Processes under Intermittent Observation. AB - In this article, we present a new statistical methodology for longitudinal studies in forestry, where trees are subject to recurrent infection, and the hazard of infection depends on tree growth over time. Understanding the nature of this dependence has important implications for reforestation and breeding programs. Challenges arise for statistical analysis in this setting with sampling schemes leading to panel data, exhibiting dynamic spatial variability, and incomplete covariate histories for hazard regression. In addition, data are collected at a large number of locations, which poses computational difficulties for spatiotemporal modeling. A joint model for infection and growth is developed wherein a mixed nonhomogeneous Poisson process, governing recurring infection, is linked with a spatially dynamic nonlinear model representing the underlying height growth trajectories. These trajectories are based on the von Bertalanffy growth model and a spatially varying parameterization is employed. Spatial variability in growth parameters is modeled through a multivariate spatial process derived through kernel convolution. Inference is conducted in a Bayesian framework with implementation based on hybrid Monte Carlo. Our methodology is applied for analysis in an 11-year study of recurrent weevil infestation of white spruce in British Columbia. PMID- 19673863 TI - Partial-likelihood analysis of spatio-temporal point-process data. AB - We investigate the use of a partial likelihood for estimation of the parameters of interest in spatio-temporal point-process models. We identify an important distinction between spatially discrete and spatially continuous models. We focus our attention on the spatially continuous case, which has not previously been considered. We use an inhomogeneous Poisson process and an infectious disease process, for which maximum-likelihood estimation is tractable, to assess the relative efficiency of partial versus full likelihood, and to illustrate the relative ease of implementation of the former. We apply the partial-likelihood method to a study of the nesting pattern of common terns in the Ebro Delta Natural Park, Spain. PMID- 19673864 TI - A bayesian approach to surrogacy assessment using principal stratification in clinical trials. AB - A surrogate marker (S) is a variable that can be measured earlier and often more easily than the true endpoint (T) in a clinical trial. Most previous research has been devoted to developing surrogacy measures to quantify how well S can replace T or examining the use of S in predicting the effect of a treatment (Z). However, the research often requires one to fit models for the distribution of T given S and Z. It is well known that such models do not have causal interpretations because the models condition on a postrandomization variable S. In this article, we directly model the relationship among T, S, and Z using a potential outcomes framework introduced by Frangakis and Rubin (2002, Biometrics 58, 21-29). We propose a Bayesian estimation method to evaluate the causal probabilities associated with the cross-classification of the potential outcomes of S and T when S and T are both binary. We use a log-linear model to directly model the association between the potential outcomes of S and T through the odds ratios. The quantities derived from this approach always have causal interpretations. However, this causal model is not identifiable from the data without additional assumptions. To reduce the nonidentifiability problem and increase the precision of statistical inferences, we assume monotonicity and incorporate prior belief that is plausible in the surrogate context by using prior distributions. We also explore the relationship among the surrogacy measures based on traditional models and this counterfactual model. The method is applied to the data from a glaucoma treatment study. PMID- 19673865 TI - Utility-based optimization of combination therapy using ordinal toxicity and efficacy in phase I/II trials. AB - An outcome-adaptive Bayesian design is proposed for choosing the optimal dose pair of a chemotherapeutic agent and a biological agent used in combination in a phase I/II clinical trial. Patient outcome is characterized as a vector of two ordinal variables accounting for toxicity and treatment efficacy. A generalization of the Aranda-Ordaz model (1981, Biometrika 68, 357-363) is used for the marginal outcome probabilities as functions of a dose pair, and a Gaussian copula is assumed to obtain joint distributions. Numerical utilities of all elementary patient outcomes, allowing the possibility that efficacy is inevaluable due to severe toxicity, are obtained using an elicitation method aimed to establish consensus among the physicians planning the trial. For each successive patient cohort, a dose pair is chosen to maximize the posterior mean utility. The method is illustrated by a trial in bladder cancer, including simulation studies of the method's sensitivity to prior parameters, the numerical utilities, correlation between the outcomes, sample size, cohort size, and starting dose pair. PMID- 19673866 TI - Semiparametric bayes multiple testing: applications to tumor data. AB - In National Toxicology Program (NTP) studies, investigators want to assess whether a test agent is carcinogenic overall and specific to certain tumor types, while estimating the dose-response profiles. Because there are potentially correlations among the tumors, a joint inference is preferred to separate univariate analyses for each tumor type. In this regard, we propose a random effect logistic model with a matrix of coefficients representing log-odds ratios for the adjacent dose groups for tumors at different sites. We propose appropriate nonparametric priors for these coefficients to characterize the correlations and to allow borrowing of information across different dose groups and tumor types. Global and local hypotheses can be easily evaluated by summarizing the output of a single Monte Carlo Markov chain (MCMC). Two multiple testing procedures are applied for testing local hypotheses based on the posterior probabilities of local alternatives. Simulation studies are conducted and an NTP tumor data set is analyzed illustrating the proposed approach. PMID- 19673867 TI - On distance-based permutation tests for between-group comparisons. AB - Permutation tests based on distances among multivariate observations have found many applications in the biological sciences. Two major testing frameworks of this kind are multiresponse permutation procedures and pseudo-F tests arising from a distance-based extension of multivariate analysis of variance. In this article, we derive conditions under which these two frameworks are equivalent. The methods and equivalence results are illustrated by reanalyzing an ecological data set and by a novel application to functional magnetic resonance imaging data. PMID- 19673868 TI - The additional value of ambulatory urodynamic measurements compared with conventional urodynamic measurements. AB - OBJECTIVES: To study the additional diagnostic value of ambulatory urodynamic measurements/monitoring (AUM) in patients with lower urinary tract symptoms (LUTS). PATIENTS AND METHODS: We reviewed the urodynamic data, collected at the urology department of our University Hospital between 2002 and 2007. During this period, 2393 urodynamic investigations were conducted. In 108 patients both conventional urodynamic measurements (CUM) and AUM were conducted. RESULTS: In 25 patients an AUM was conducted for bladder evacuation problems due to absent bladder contractility, seen on CUM. In 21 cases, AUM showed the presence of contractility of the bladder under normal conditions at home. Their symptoms were due to other factors such as a concomitant non-relaxation of the urinary sphincter or pelvic floor, psychological reasons or obstruction, for all of which treatment could be initiated. In 32 cases, the indication for AUM was an inconclusive CUM. Of these patients 16 had clear overactive contractions on AUM. CONCLUSION: AUM has a more important place in the second-line diagnostic evaluation of patients with LUTS than generally considered. In half of the cases (16 of 32) in which CUM could not provide a diagnosis, AUM helped us to diagnose overactive bladder. Moreover, in the absence of AUM, many patients would have been misdiagnosed with an acontractile bladder based on their CUM results, which would most probably have resulted in life-time clean intermittent self catheterization. PMID- 19673869 TI - Stretch independent regulation of prostaglandin E(2) production within the isolated guinea-pig lamina propria. AB - OBJECTIVE: To use an isolated preparation of the guinea-pig bladder lamina propria (LP) to investigate the effects of adenosine tri-phosphate (ATP) and nitric oxide (NO) on the release of prostaglandin E(2) (PGE(2)). MATERIALS AND METHODS: The bladders of female guinea-pigs (200-400 g) were isolated and opened to expose the urothelial surface. The LP was dissected free of the underlying detrusor muscle and cut into strips from the dome to base. Strips were then incubated in Krebs buffer at 37 degrees C. Each tissue piece was then exposed to the stable ATP analogue, BzATP, and a NO donor, diethylamine-NONOate (DEANO), and the effect on PGE(2) output into the supernatant determined using the Parameter(TM) PGE(2) enzyme immunoassay kit (R & D Systems, Abingdon, UK). Experiments were repeated in the presence of purinergic receptor and cyclooxygenase (COX) enzymes, COX I and COX II, antagonists. The cellular location of COX I, COX II and neuronal NO synthase (nNOS) within the bladder LP was also determined by immunohistochemistry. RESULTS: PGE(2) production was significantly increased by BzATP. Antagonist studies showed the purinergic stimulation involved both P(2)X and P(2)Y receptors. The BzATP response was inhibited by the COX inhibitor indomethacin (COX I >COX II) but not by DUP 697 (COX II >COX I). Thus, BzATP stimulation occurs because of COX I stimulation. NO had no effect on PGE(2) production over the initial 10 min of an exposure. However, PGE(2) output was increased 100 min after exposure to the NO donor. In the presence of NO, the BzATP stimulation was abolished. Immunohistochemistry was used to confirm the location of COX I to the basal and inner intermediate urothelial layers and to cells within the diffuse layer of LP interstitial cells. In addition, nNOS was also located in the basal urothelial layers whilst COX II was found in the interstitial cell layers. CONCLUSIONS: There is complex interaction between ATP and NO to modulate PGE(2) release from the bladder LP in the un-stretched preparation. Such interactions suggest a complex interrelationship of signals derived from this region of the bladder wall. The importance of these interactions in relation to the physiology of the LP remains to be determined. PMID- 19673870 TI - MYC gene numerical aberrations in actinic keratosis and cutaneous squamous cell carcinoma. AB - BACKGROUND: The genetic alterations that drive the transition from actinic keratoses (AKs) to cutaneous squamous cell carcinomas (SCCs) have not been defined precisely. Amplification and/or overexpression of the MYC proto-oncogene have been demonstrated in several human, malignant tumours including head and neck SCCs. OBJECTIVES: To evaluate the presence of MYC genomic aberrations in both AKs and SCCs. METHODS: Skin biopsy specimens corresponding to AKs, SCCs and control samples were included in two paraffin-embedded tissue microarrays. MYC cytogenetic profile was evaluated by fluorescence in situ hybridization (FISH). The results obtained were compared with MYC immunohistochemical expression. RESULTS: Twenty-three AKs and 30 SCCs were evaluated. MYC numerical aberrations were observed in eight of 23 (35%) AKs and 19 of 30 (63%) SCCs (P = 0.05). MYC numerical aberrations were more frequent in moderately to poorly differentiated SCCs (77%) when compared with well-differentiated SCCs (25%; P = 0.027). A significant association between copy number gains of MYC by FISH analysis and MYC protein expression was demonstrated. CONCLUSIONS: MYC gains and amplifications are frequent cytogenetic abnormalities in SCCs and may play a relevant role in promoting SCC undifferentiation and tumoral progression. PMID- 19673871 TI - Nicotine enhances skin necrosis and expression of inflammatory mediators in a rat pressure ulcer model. AB - BACKGROUND: Many bedridden patients develop pressure ulcers, not only in hospital but also at home. Clinical studies have indicated cigarette smoking to be a risk factor for pressure ulcers. However, the contribution of nicotine to pressure ulcer formation has not been identified. OBJECTIVES: We aimed to clarify the effect of nicotine on pressure ulcer formation, and its mechanism. METHODS: Ischaemia-reperfusion (I/R) was performed in rat dorsal skin to induce pressure ulcers. The extent of the resulting necrotic area was determined. To clarify the mechanism of the effect of nicotine, mRNA levels of cyclooxygenase-2 (COX-2), interleukin (IL)-1beta, IL-6 and inducible nitric oxide synthase (iNOS) and protein expression of COX-2 and iNOS in the necrotic area were investigated by real-time reverse transcription-polymerase chain reaction and Western blotting, respectively. Furthermore, the effects of the COX-2 inhibitor NS-398 and the iNOS inhibitor aminoguanidine on necrosis were examined. RESULTS: Skin necrosis in the I/R-treated area was significantly increased by intraperitoneal administration of nicotine (0.175 mg kg(-1) daily). Repeated nicotine administration had little effect on systolic and diastolic blood pressure. I/R treatment increased mRNA levels of COX-2, IL-1beta, IL-6 and iNOS, which were further augmented by nicotine in a dose-dependent manner. Correspondingly, nicotine (0.35 mg kg(-1) daily) markedly enhanced the protein expression of COX-2 and iNOS. Moreover, NS 398 and aminoguanidine showed a tendency to abrogate the increase of I/R-induced skin necrosis caused by nicotine. CONCLUSIONS: These results suggest that the increased risk of pressure ulcers due to cigarette smoking is mediated, in part, by nicotine. They also indicated that the effect of nicotine is not mediated by a change in blood pressure, but is elicited via an increase of inflammatory mediators in the I/R-treated skin. PMID- 19673872 TI - A novel missense mutation in tumour necrosis factor receptor superfamily 1A (TNFRSF1A) gene found in tumour necrosis factor receptor-associated periodic syndrome (TRAPS) manifesting adult-onset Still disease-like skin eruptions: report of a case and review of the Japanese patients. PMID- 19673873 TI - A homozygous mutation in the ferrochelatase gene underlies erythropoietic protoporphyria associated with palmar keratoderma--reply. PMID- 19673874 TI - The prevalence of chromium allergy in Denmark is currently increasing as a result of leather exposure. AB - BACKGROUND: Chromium allergy has traditionally been caused by occupational skin contact with cement. In 1983, Danish legislation made the addition of ferrous sulphate compulsory in cement to reduce the water-soluble chromium content to not more than 2 ppm. An effect from this intervention has previously been demonstrated among Danish construction workers. OBJECTIVES: To investigate the development of chromium allergy among patients with dermatitis tested between 1985 and 2007 in Denmark. Furthermore, to determine causative exposures in patients with chromium allergy. PATIENTS AND METHODS: A retrospective analysis of patch test data was performed (n = 16,228) and charts from patients with chromium allergy were reviewed. Comparisons were made using a chi(2) test. Logistic regression analyses were used to test for associations. RESULTS: The prevalence of chromium allergy decreased significantly from 3.6% in 1985 to 1% in 1995 (P(trend) < 0.001) but increased to 3.3% in 2007 (P(trend) < 0.001). The frequency of clinically relevant cement exposure decreased significantly among patients with chromium allergy from 12.7% in 1989-1994 to 3.0% (P < 0.01) in 1995 2007, whereas the frequency of relevant leather exposure increased significantly from 24.1% during 1989-1994 to 45.5% during 1995-2007 (P < 0.02). CONCLUSIONS: Chromium allergy is currently increasing in Denmark due to leather exposure. PMID- 19673875 TI - Targeted treatment of pyoderma gangrenosum in PAPA (pyogenic arthritis, pyoderma gangrenosum and acne) syndrome with the recombinant human interleukin-1 receptor antagonist anakinra. AB - The triad of sterile pyogenic arthritis, pyoderma gangrenosum and acne is known by the acronym of PAPA syndrome. It is a rare autosomal dominant disease of early onset. The treatment of pyoderma gangrenosum is challenging as there is often only partial response to systemic glucocorticosteroids and immunosuppressive therapies. We report the rapid and lasting response of pyoderma gangrenosum to the targeted treatment with the recombinant human interleukin-1 receptor antagonist (rHuIL-1Ra) anakinra in a patient with PAPA syndrome. PMID- 19673876 TI - Closure of dehisced operative sites without wound freshening results in acceptable rates of repeat dehiscence and infection. PMID- 19673877 TI - The familial feature of Tunisian endemic pemphigus foliaceus. PMID- 19673878 TI - Long-term results of 2-mm punch grafting in patients with vitiligo vulgaris and segmental vitiligo: effect of disease activity. AB - BACKGROUND: Punch grafting is a simple and frequently used technique for the treatment of stable vitiligo, resistant to medical therapy. However, studies reporting long-term results are exceptional. OBJECTIVES: To evaluate the long term results of 2-mm punch grafting in patients with vitiligo vulgaris and segmental vitiligo. METHODS: We studied a prospective cohort study involving 61 patients (25 male, 36 female) with vitiligo vulgaris and nine patients (all male) with segmental vitiligo who underwent 2-mm punch grafting more than 3 years ago. The main outcome measure was the degree of repigmentation of a single transplanted lesion as measured with a digital image analysis system with a mean follow-up of 5.2 years. RESULTS: In patients with vitiligo vulgaris, 17 lesions (28%) showed excellent, 14 lesions (23%) showed good, 14 lesions (23%) showed fair and 16 lesions (26%) showed poor repigmentation. In patients with segmental vitiligo, seven of nine lesions (78%) showed excellent repigmentation. A cobblestone-like effect was observed in 19 of 70 patients (27%). Disease activity after punch grafting was reported in 94% of patients with poor repigmentation but in only 18% of patients with excellent repigmentation (chi(2) test, P < 0.0005). Patients who reported disease activity after transplantation had a lower mean repigmentation than those who did not report disease activity (77% vs. 39%, P < 0.05). CONCLUSIONS: Two-millimetre punch grafting in vitiligo is an effective surgical procedure with long-lasting effect. To prevent a cobblestone-like effect, we advise the use of smaller grafts (1-1.2 mm). Disease activity after grafting, localization and type of vitiligo, prior ultraviolet B treatment and a Koebnerized donor site influence the long-term outcome of punch grafting and should be taken into account in the selection of patients eligible for this treatment. PMID- 19673879 TI - Low bone mineral density in adult patients with moderate to severe atopic dermatitis. AB - BACKGROUND: Atopic dermatitis (AD) is a chronic inflammatory skin disease commonly treated with topical corticosteroids. The inflammatory nature of this disorder and the use of topical corticosteroids represent potential risk factors for bone loss. OBJECTIVES: The aim was to assess the prevalence of osteoporosis and osteopenia in adult patients with moderate to severe AD. In addition, the associations between topical/oral corticosteroid use and bone mineral density (BMD) and between disease activity and BMD were studied. PATIENTS AND METHODS: We studied 125 adult patients with moderate to severe AD. Using dual-energy X-ray absorptiometry, BMD was measured at lumbar spine and hips. The cumulative dose of topical and oral corticosteroids was calculated from pharmacy prescription records. Lifestyle parameters were collected by a questionnaire. Biochemical parameters of bone metabolism and disease activity [serum concentration of thymus and activation-regulated chemokine (TARC) levels] were also measured. RESULTS: Osteoporosis was documented in six patients (4.8%) and osteopenia in 41 patients (32.8%); 30.4% of the patients had a Z-score 0.05). There were no changes in VIP-, NPY- or somatostatin-LI. In conclusion, the NO donor GTN appears not to induce headache via immediate CGRP release. PMID- 19673899 TI - Extracranial source of increased CGRP in migraine children? PMID- 19673904 TI - Ophthalmoplegic migraine: the case for reclassification. PMID- 19673907 TI - MTHFR 677C->T and ACE D/I polymorphisms and migraine attack frequency in women. AB - Data on the association of the MTHFR 677C->T and ACE D/I polymorphisms with migraine severity, measured by attack frequency, are scarce. We performed an association study among 24 961 women participating in the Women's Health Study. Migraine, aura status and attack frequency were self-reported. Multinomial logistic regression was used to investigate the genotype-migraine association. Among the 3186 migraineurs with complete genotype and attack frequency data, 1270 reported migraine with aura (MA) (attack frequency 76 >= weekly; 219 monthly; 123 every other month; 852 fewer than six times/year) and 1916 migraine without aura (MoA) (attack frequency: 85 >= weekly; 414 monthly; 208 every other month; 1209 fewer than six times/year). The MTHFR 677TT genotype was associated with a reduced risk for MA, which only appeared for attacks fewer than six times/year (age-adjusted odds ratio 0.78; 95% confidence interval 0.61, 0.99). We did not find a specific pattern of association of the ACE D/I polymorphism with attack frequency for MA or MoA. PMID- 19673908 TI - Clinical and neuroimaging evidence of interictal cerebellar dysfunction in FHM2. AB - We used multimodal magnetic resonance (MR) techniques [brain diffusion-weighted magnetic resonance imaging, diffusion-weighted imaging (DWI), proton MR spectroscopy (MRS), (1)H-MRS; and skeletal muscle phosphorous MRS, (31)P-MRS] to investigate interictal brain microstructural changes and tissue energy metabolism in four women with genetically determined familial hemiplegic migraine type 2 (FHM2), belonging to two unrelated families, compared with 10 healthy women. Brain DWI revealed a significant increase of the apparent diffusion coefficient median values in the vermis and cerebellar hemispheres of FHM2 patients, preceding in two subjects the onset of interictal cerebellar deficits. (31)P-MRS revealed defective energy metabolism in skeletal muscle of FHM2 patients, while brain (1)H-MRS showed a mild pathological increase in lactate in the lateral ventricles of one patient and a mild reduction of cortical N-acetyl-aspartate to creatine ratio in another one. Our MRS results showed that a multisystem energy metabolism defect in FHM2 is associated with microstructural cerebellar changes detected by DWI, even before the onset of cerebellar symptoms. PMID- 19673909 TI - Migraine with atypical visual aura caused by colloid cyst of the third ventricle. PMID- 19673910 TI - Incidence and predictors of chronic headache attributed to whiplash injury. AB - We identified clinical, demographic and psychological predictive factors that may contribute to the development of chronic headache associated with mild to moderate whiplash injury [Quebec Task Force (QTF) <= II] and determined the incidence of this chronic pain state. Patients were recruited prospectively from six participating accident and emergency departments. While 4.6% of patients developed chronic headache attributed to whiplash injury according to the International Classification of Headache Disorders, 2nd edn criteria, 15.2% of patients complained about headache lasting > 42 days (QTF criteria). Predictive factors were pre-existing facial pain [odds ratio (OR) 9.7, 95% confidence interval (CI) 2.1, 10.4; P = 0.017], lack of confidence to recover completely (OR 5.5, 95% CI 2.0, 13.2; P = 0.005), sore throat (OR 5.0, 95% CI 1.5, 8.9; P = 0.013), medication overuse (OR 4.2, 95% CI 1.4, 12.3; P = 0.009), high Neck Disability Index (OR 4.0, 95% CI 1.3, 12.6; P = 0.019), hopelessness/anxiety (OR 3.8, 95% CI 1.3, 8.7; P = 0.024), and depression (OR 3.3, 95% CI 1.2, 9.4; P = 0.024). The lack of a control group limits the conclusions that can be drawn from this study. Identified predictors closely resemble those found in chronic primary headache disorders. PMID- 19673911 TI - Methylprednisolone therapy for short-term prevention of SUNCT syndrome. AB - Short-lasting unilateral neuralgiform headache attacks with conjunctival injection and tearing (SUNCT) is characterized by severe and frequent daily pain attacks causing transient physical disability for the patients during the headache period. Currently there is no option for abortive treatment of the attacks, mainly due to the short-lived nature and frequency of the repeated headaches, while highly efficacious therapy is also unavailable for short-term prevention. We report rapidly suppressed headache attacks with orally administered methylprednisolone in eight headache periods of three patients with idiopathic, episodic SUNCT syndrome. The remission was maintained until the period was over in all cases. Although the mechanism of methylprednisolone action is unclear, it is probably based on the anti-inflammatory effects of the drug. PMID- 19673912 TI - Headache attributed to intracranial tumours: a prospective cohort study. AB - Between January 2007 and March 2008, we prospectively studied all patients operated on for intracranial tumours in our Department of Neurosurgery. Preoperatively, all patients were interviewed by a neurologist to collect headache characteristics. Measurements of tumour and oedema volume were made using dedicated software for magnetic resonance imaging studies. Tumour histopathology was established by histological examination postoperatively. If headache improved postoperatively, a diagnosis of 'headache attributed to intracranial neoplasm' was made, according to the 2004 International Classification of Headache Disorders (ICHD-II). A multivariate logistic regression model was used to evaluate the association of headache with potential risk factors. We studied 206 subjects. The prevalence of tumour headache was 47.6%. Intracranial tumour headache was non-specific and in most cases could not be classified by current ICHD-II diagnostic criteria for primary headache syndromes. Its prevalence varied depending on volume, location and type of tumour, as well as on the patient's previous headache history. PMID- 19673913 TI - Epidemiological, clinical and pharmacological aspects of headache in a university undergraduate population in Palestine. AB - Headache is one of the most common complaints in clinical practice. Few studies regarding headache in university students have been conducted in the Middle East. The objective of this study was to explore the prevalence, clinical characteristics, triggering factors and treatment options of headaches in university undergraduate students in Palestine/Middle East. Data were collected by interviewing a sample of 1900 students. The Headache Assessment Quiz was used to measure quality and severity of headache and to collect data on triggering factors and symptom management. A total of 1808 (95.2%) reported having at least one headache episode in the previous year. A positive family history of headache was found in 40% of students. The prevalence rate of frequent headache (tow or more episodes/month) was found in 1096 (60.9%) students; 613 women (55.9%). Of those having frequent headaches, 228 (20.8%) experienced moderate to severe episodes, 341 (31.2%) had pulsating, throbbing and pounding pain, and 274 (25%) had unilateral pain. The most common triggering factors among students with frequent headaches were: tension/stress (78.2%) and sleep deprivation (75.4%). Less than 5% of students sought medical assistance during headache episodes. Most students (79.1%) reported self-therapy with a single analgesic (53.4%), herbs (10.2%) or combination (15.5%), while 20.9% reported using no medication of any type to decrease pain. Paracetamol (48.5%) followed by ibuprofen (4.9%) were the most commonly used non-prescription analgesic drugs. Headache is a prevalent symptom in the college age population. Further research is needed to determine the prevalence of specific types of headaches. Healthcare providers are required to educate this population as well as to assist students in properly diagnosing and treating headache types. PMID- 19673914 TI - A new trigemino-nociceptive stimulation model for event-related fMRI. AB - Functional imaging of human trigemino-nociceptive processing provides meaningful insights into altered pain processing in head and face pain diseases. Although functional magnetic resonance imaging (fMRI) offers high temporal and spatial resolution, most studies available were done with radioligand-positron emission tomography, as fMRI requires non-magnetic stimulus equipment and fast on-off conditions. We developed a new approach for painful stimulation of the trigeminal nerve that can be implemented within an event-related design using fMRI and aimed to detect increased blood-oxygen-level-dependent (BOLD) signals as surrogate markers of trigeminal pain processing. Using an olfactometer, 20 healthy volunteers received intranasally standardized trigeminal nociceptive stimuli (ammonia gas) as well as olfactory (rose odour) and odorless control stimuli (air puffs). Imaging revealed robust BOLD responses to the trigeminal nociceptive stimulation in cortical and subcortical brain areas known to be involved in pain processing. Focusing on the trigeminal pain pathway, significant activations were observed bilaterally in brainstem areas at the trigeminal nerve entry zone, which are agreeable with the principal trigeminal nuclei. Furthermore, increased signal changes could be detected ipsilaterally at anatomical localization of the trigeminal ganglion and bilaterally in the rostral medulla, which probably represents the spinal trigeminal nuclei. However, brainstem areas involved in the endogenous pain control system that are close to this anatomical localization, such as raphe nuclei, have to be discussed. Our findings suggest that mapping trigeminal pain processing using fMRI with this non-invasive experimental design is feasible and capable of evoking specific activations in the trigeminal nociceptive system. This method will provide an ideal opportunity to study the trigeminal pain system in both health and pathological conditions such as idiopathic headache disorders. PMID- 19673915 TI - Role of the oestrogen receptor (ESR1 PvuII and ESR1 325 C->G) and progesterone receptor (PROGINS) polymorphisms in genetic susceptibility to migraine in a North Indian population. AB - We aimed to explore the single-locus, haplotype and epistasis patterns and the contribution of oestrogen receptor [ESR1 PvuII (rs2234693), ESR1 325 C->G (rs1801132)] and progesterone receptor [PROGINS (rs1042838)] polymorphisms in genetic susceptibility to migraine by analysing 613 subjects consisting of 217 migraine patients, 217 healthy controls (HC) and 179 patients with tension-type headache (TTH). Entire data were analysed by taking the Bonferroni corrected P value into account. We found significant association of TT genotype [odds ratio (OR) 3.458, confidence interval (CI) 1.757, 6.806; P = 0.0003] and T allele (OR 1.729, CI 1.309, 2.284; P = 0.0001) of ESR1 PvuII single nucleotide polymorphism with migraine when compared with HC. Significant association was seen only in female migraine patients at both genotype (P = 0.002; OR 3.834, CI 1.625, 9.043) and allele level (P = 0.002; OR 1.721, CI 1.228, 2.413). Moreover, higher risk was limited to migraine with aura (MA) (in case of TT genotype, P = 0.002; OR 4.377, CI 1.703, 1.246; in case of T allele, P = 0.001; OR 1.888, CI 1.305, 2.735) rather than migraine without aura (MoA) (P-value of TT genotype = 0.003; OR 3.082, CI 1.465, 6.483; P-value T allele = 0.002; OR 1.630, CI 1.188, 2.236). In case of a recessive model, risk was seen with migraine patients (P = 0.0003; OR 2.514, CI 1.635, 3.867), MA (P = 0.0001; OR 3.583, CI 1.858, 6.909) and MoA patients (P = 0.002; OR 2.125, CI 1.304, 3.464) when compared with HC. No risk was observed when TTH patients were compared with HC. No significance of ESR 325 G->C polymorphism was seen in any of the models under study. Significant differences in genotypic (P = 0.0001) and allelic frequency (P = 0.0002) were seen in case of PROGINS polymorphism when migraine patients were compared with HC, showing a protective effect (for A1A2 genotype, OR 0.292, CI 0.155, 0.549; for A2 allele, OR 0.320, CI 0.174, 0.589). Moreover, significance was seen only in case of female migraine patients at genotype (P = 0.002; OR 0.344, CI 0.176, 0.684) as well as allele levels (P = 0.004; OR 0.379, CI 0.198, 0.727) in case of PROGINS polymorphism. ESR1 PvuII TT*ESR1 325 C->G CG genotype, PROGINS A1A2*ESR1 325 C->G CG genotype and ESR1 PvuII CT*PROGINS A1A2 interacted significantly, but significance was lost after Bonferroni correction. In conclusion, ESR1 PvuII polymorphism is a significant risk factor for migraine particularly in women and MA patients, but ESR 325 C->G polymorphism is not associated with migraine susceptibility. PROGINS polymorphism seems to play a protective role in genetic susceptibility to migraine in the North Indian population. PMID- 19673916 TI - A survey of family doctors on the likeability of migraine and other common diseases and their prevalence of migraine. AB - A survey of 148 family doctors attending a continuing medical education migraine update lecture was performed to assess whether family doctors like to treat migraine and other common disorders and the prevalence of migraine. Doctors were asked to respond to the following statement using a five-point Likert scale (from 1, strongly disagree to 5, strongly agree): 'I like to treat patients with this disease or symptom'. The response rate was 53% with a mean age of 51.5 years. Doctors reported liking to treat general medical conditions more (mean = 4.40) than migraine (mean = 3.38) and other neurological diseases (mean = 3.20). Doctors reported a personal history of migraine in the prior 1 year of 22.8% and 45.6% lifetime, with 17% becoming aware for the first time that they personally had migraine after attending the lecture. Respondents with a personal history of migraine liked to treat migraine more than those without a history. PMID- 19673918 TI - The future of headache classification and classification research. PMID- 19673917 TI - Proposals for new standardized general diagnostic criteria for the secondary headaches. AB - Headache classification is a dynamic process through clinical testing and re testing of current and proposed criteria. After publication of the second edition of the International Classification of Headache Disorders (ICHD-II), need arose for revisions in the classification of medication overuse headache and chronic migraine. These changes made apparent a further need for broader revisions to the standard formulation of diagnostic criteria for the secondary headaches. Currently, the fourth criterion makes impossible the definitive diagnosis of a secondary headache until the underlying cause has resolved or been cured or greatly ameliorated by therapy, at which time the headache may no longer be present. Given that the main purpose of diagnostic criteria is to enable a diagnosis at the onset of a disease in order to guide treatment, this is unhelpful in clinical practice. In the present paper we propose maintaining a standard approach to the secondary headaches using a set of four criteria A, B, C and D, but we construct these so that the requirement for resolution or successful treatment is removed. The proposal for general diagnostic criteria for the secondary headaches will be entered into the internet-based version of the appendix of ICHD-II. During 2009 the Classification Committee will apply the general criteria to all the specific types of secondary headaches. These, and other changes, will be included in a revision of the entire classification entitled ICHD-IIR, expected to be published in 2010. ICHD-IIR will be printed and posted on the website and will be the official classification of the International Headache Society. Unfortunately, it will be necessary to translate ICHD-IIR into the many languages of the world, but the good news is that no major changes to the headache classification are then foreseen for the next 10 years. Until the printing of ICHD-IIR, the printed ICHD-II criteria remain in place for all other purposes. We issue a plea to the headache community to use and study these proposed general criteria for the secondary headaches in order to provide more evidence for their utility-before their incorporation in the main body of the classification. PMID- 19673919 TI - Clinical experience of TiUnite implants: a 5-year cross-sectional, retrospective follow-up study. AB - BACKGROUND: Little is known of the long-term clinical and radiographic performance of moderately rough surface implants. PURPOSE: The aim of the present retrospective investigation was to study two pioneer cohorts of patients, that is, the first patients to receive Branemark System implants with a moderately rough surface (TiUnite, Nobel Biocare AB, Goteborg, Sweden) at the present clinic. TiUnite implants were inserted either in compromised bone sites in a mixed-mouth concept together with turned implants or used solely. Patients were followed up over a period of 5 years with regard to implant survival and the marginal bone response. MATERIALS AND METHODS: Patients who received both implant types (mixed group) comprised 41 subjects, and the second group (TiUnite group) comprised 70 subjects. A total of 110 turned and 68 TiUnite implants were placed in the mixed group, and 212 TiUnite implants in the TiUnite group. Follow-up radiographs were obtained at prosthesis placement and at the 1- and 5-year check ups, and examined by independent observers. RESULTS: One turned (0.9%) and two TiUnite (2.9%) implants failed in the mixed group, and three implants (1.6%) failed in the TiUnite group, indicating no significant differences between surfaces or groups (p < .05). The mean marginal bone loss at 5 years was 0.6 mm to 0.8 mm, also indicating no significant differences for the two implant types tested in the mixed group. CONCLUSIONS: Cumulative survival rates for the two implant surfaces were favorable at 5 years, and the marginal bone loss was low and similar for both implant surfaces. PMID- 19673920 TI - The peri-implant sulcus compared with internal implant and suprastructure components: a microbiological analysis. AB - PURPOSE: A recent in vivo study has shown considerable contamination of internal implant and suprastructure components with great biodiversity, indicating bacterial leakage along the implant-abutment interface, abutment-prosthesis interface, and restorative margins. The goal of the present study was to compare microbiologically the peri-implant sulcus to these internal components on implants with no clinical signs of peri-implantitis and in function for many years. Checkerboard DNA-DNA hybridization was used to identify and quantify 40 species. MATERIAL AND METHODS: Fifty-eight turned titanium Branemark implants in eight systemically healthy patients (seven women, one man) under regular supportive care were examined. All implants had been placed in the maxilla and loaded with a screw-retained full-arch bridge for an average of 9.6 years. Gingival fluid samples were collected from the deepest sulcus per implant for microbiological analysis. As all fixed restorations were removed, the cotton pellet enclosed in the intra-coronal compartment and the abutment screw were retrieved and microbiologically evaluated. RESULTS: The pellet enclosed in the suprastructure was very similar to the peri-implant sulcus in terms of bacterial detection frequencies and levels for practically all the species included in the panel. Yet, there was virtually no microbial link between these compartments. When comparing the abutment screw to the peri-implant sulcus, the majority of the species were less frequently found, and in lower numbers at the former. However, a relevant link in counts for a lot of bacteria was described between these compartments. Even though all implants in the present study showed no clinical signs of peri-implantitis, the high prevalence of numerous species associated with pathology was striking. CONCLUSIONS: Intra-coronal compartments of screw retained fixed restorations were heavily contaminated. The restorative margin may have been the principal pathway for bacterial leakage. Contamination of abutment screws most likely occurred from the peri-implant sulcus via the implant-abutment interface and abutment-prosthesis interface. PMID- 19673921 TI - Efficacy of cancellous block allograft augmentation prior to implant placement in the posterior atrophic mandible. AB - BACKGROUND: The present study evaluated the outcome of ridge augmentation with cancellous freeze-dried block bone allografts in the posterior atrophic mandible followed by placement of dental implants. MATERIALS AND METHODS: A bony deficiency of at least 3 mm, horizontally, vertically, or both, according to computerized tomography (CT) para-axial reconstruction served as inclusion criteria. Implants were inserted after a healing period of 6 months. Bone measurements were taken prior to bone augmentation, during implant placement, and at second-stage surgery. Marginal bone loss and crown-to-implant ratio were also measured. RESULTS: Twenty-nine cancellous allogeneic bone blocks were placed in 21 patients. The mean follow-up was 37 months. Bone block survival rate was 79.3%. Mean horizontal and vertical bone gains were 5.6 and 4.3 mm, respectively. Mean buccal bone resorption was 0.5 mm at implant placement and 0.2 mm at second stage surgery. A total of 85 implants were placed. Mean bone thickness buccal to the implant neck was 2.5 mm at implant placement and 2.3 mm at second-stage surgery. There was no evidence of vertical bone loss between implant placement and second-stage surgery. Implant survival rate was 95.3%. All patients received a fixed implant-supported prosthesis. At the last follow-up, the mean marginal bone loss was 0.5 mm. The mean crown-to-implant ratio was 0.96. CONCLUSION: Implant placement in the posterior atrophic mandible following augmentation with cancellous freeze-dried bone block allografts may be regarded as a viable treatment alternative. PMID- 19673922 TI - Early versus delayed loading of mandibular implant-supported overdentures: 5-year results. AB - BACKGROUND AND PURPOSE: Because of poor retention of complete removable dentures for edentulous patients, implant-supported mandibular overdentures have lately become a popular alternative for them. The aims of this prospective study were to evaluate treatment outcomes of mandibular overdentures supported by two unsplinted early-loaded implants and compare these results with those for delayed loaded implants. MATERIALS AND METHODS: A total of 26 edentulous patients were treated with two unsplinted implants supporting a mandibular overdenture. All implants were placed in the canine regions of each mandible according to the one stage surgical protocol. There were two groups: test group, in which the overdenture was connected 1 week after surgery, and control group, in which the overdenture was connected 3 months after surgery. Standardized clinical and radiographic parameters were recorded at surgery, and after 3, 6, 12, and 18 months, and 2, 3, 4, and 5 years. RESULTS: No implants were lost, and 0.93 +/- 0.3 mm marginal bone resorption was noted for all implants after 5 years. Clinical implant stability measurements, clinical peri-implant parameters, and marginal bone resorptions showed no statistically significant differences between the two groups over 5 years. CONCLUSION: The results of this prospective clinical study suggest that there is no significant difference in the clinical and radiographic state of patients treated with implant supported mandibular overdentures loaded either 1 week or 3 months after surgery. PMID- 19673923 TI - Effects of flapless implant surgery on soft tissue profiles: a prospective clinical study. AB - BACKGROUND: Flapless implant surgery has been suggested as a suitable treatment modality for the preservation of soft tissue after implant placement. PURPOSE: The purpose of this study was to determine the extent of soft tissue profile changes around implants after flapless implant surgery. MATERIALS AND METHODS: A total of 44 patients received 76 implants using a flapless implant procedure. The marginal level of the peri-implant soft tissue was evaluated using dental casts 1 week, 1 month, and 4 months after implant placement. RESULTS: The mean soft tissue levels around implants showed 0.7 +/- 0.3 mm of coronal growth 1 week after surgery. At 1 month, the levels were 0.2 +/- 0.2 mm coronal growth and at 4 months, the values were 0.0 +/- 0.3 mm. Soft tissue profiles assessed 4 months after flapless implant placement were similar to profiles assessed immediately before implant placement. CONCLUSION: Flapless implant surgery is advantageous for preserving mucosal form surrounding dental implants. PMID- 19673924 TI - The influence of surface roughness on the displacement of osteogenic bone particles during placement of titanium screw-type implants. AB - BACKGROUND: Previously, we demonstrated that bone debris, which is translocated during dental implant placement, has osteogenic potential. Therefore, it was hypothesized that implant surface roughness can influence the amount of translocated bone debris/particles and thereby the osteogenic response. MATERIAL AND METHODS: Small titanium implants were left turned (smooth) or blasted and acid etched. The implants were placed in fresh cadaver bone. After explantation, the implants were incubated in a culture medium containing beta-glycerophosphate and dexamethasone up to 24 days. Subsequently, histology, scanning electron microscopy (SEM), DNA analysis, and calcium (Ca) content measurements were performed. RESULTS: For both types of implant during implant placement, bone particles were translocated because of inherent roughness of the implant. SEM and histology confirmed the presence of a bone-like tissue on the surface of both types of implants, as also confirmed by DNA and Ca measurements. However, the significantly higher roughness of the etched implants accounted for more bone debris and accordingly elevated osteogenic response. Control samples, which had not been placed into bone, did not show mineralization in the same medium. CONCLUSION: The present study, for the first time, demonstrated that implant surface roughness can increase the amount of the translocated bone particles and thereby also have a beneficial effect on the osteogenic response of these bone particles. It is hypothesized that these bone fragments behave like miniature auto-grafts and thereby play a significant role to enhance peri-implant osteogenesis. Optimization of surface topography should be evaluated to take advantage of this additional effect of surface roughness. PMID- 19673925 TI - Perceptual changes in the peri-implant soft tissues assessed by directional cutaneous kinaesthesia and graphaesthesia: a prospective study. AB - BACKGROUND: The innervation of skin and oral mucosa plays a major physiological role in exteroception. This innervation is also clinically relevant as sensory changes occur after neurosurgical procedures. PURPOSE: The goal of this study was to compare the perception of mechanical stimuli applied to the buccal mucosa in the vicinity of osseointegrated oral implants with that in the controlateral dentate side. The role of the previously reported increased innervation in the peri-implant soft tissues in the oral sensorimotor function was thus examined. MATERIALS AND METHODS: Seventeen subjects with 20 implants were tested. Directional cutaneous kinaesthesia (DCK) and graphesthesia (G) were performed on the buccal side of the alveolar mucosa before and at planned intervals after implant placement. The observation was pursued until 6 months after the prosthetic rehabilitation. In each subject, the contralateral mucosa served as a control to the implant sites. Average percentages of correct responses in a four choice task for DCK and a three-choice task for G were calculated. RESULTS: Despite an intersubject variation in both the DCK and G, high intraindividual correlations were found (p < .005). The implant sites showed a significant difference toward the control sites at the four interval test for both tests. For DCK and G, the average of correct responses decreased after abutment connection (i.e., after the implant uncovering surgery) to increase afterwards to reach a level close to, but still lower than, the control sites 3 to 6 months after the prosthetic rehabilitation. CONCLUSION: The DCK and G are simple but reliable sensory tests that can be easily applied in the oral region. This prospective study indicates that tooth loss reduces tactile function compared with implant supported prostheses. The peri-implant soft tissues could be partially involved in the osseoperception function. PMID- 19673926 TI - Immediate occlusal loading of single lower molars using Branemark System(r) Wide Platform TiUniteTM implants: a 5-year follow-up report of a prospective clinical multicenter study. AB - INTRODUCTION: Although not essential, molars hold their importance in terms of functional jaw stability, antagonist opposition, and support of facial height. Therefore, implant therapy is an attractive concept in molar areas. However, especially in the posterior mandible, the conventional two-stage surgical approach to implant therapy was reported to cause higher bone loss and/or higher implant failures with machined implants because of the peculiar anatomic and physiologic conditions of this area. As the TiUniteTM (Nobel Biocare AB, Goteborg, Sweden) surface results in faster bone healing than with machined surface implants, it was hypothesized that this surface would also improve the performance of wide implants in posterior mandibles. Based on these assumptions, a protocol for immediately loaded implants for single molar replacement was developed. PURPOSE: This paper aimed to report on the clinical and radiological performance of Branemark System(r) TiUnite Wide Platform implants supporting single molars in the lower jaw, loaded immediately and followed for up to 5 years, and to assess if the benefit delivered by oxidized surfaces in the short run is also present after 5 years. MATERIALS AND METHODS: The study includes 33 consecutive patients treated between March 2001 and September 2003 and monitored until September 2008 in two private dental offices. A total of 40 Branemark System TiUnite Wide Platform MK III implants were placed. All implants were provided with provisional crowns in full centric occlusion at the time of surgery. Patients were clinically and radiologically followed up for up to 5 years. RESULTS: Two implant failed so that the cumulative success rate at 5 years was 95.0%. The mean marginal bone remodeling (n = 38) expressed as mesial plus distal value averages was -1.17 mm (SD +/- 0.90) at the 5-year time point. CONCLUSION: Although limited by the number of patients treated in accordance with the protocol described, 5-year results encourage the use of immediately loaded single lower molars supported by Branemark System Wide Platform TiUnite implants and further document the clinical advantages of titanium oxidized surfaces. PMID- 19673927 TI - Low density lipoprotein receptor-related protein 5 (LRP5) mutations and osteoporosis, impaired glucose metabolism and hypercholesterolaemia. AB - OBJECTIVE: Mutations in the low-density lipoprotein receptor-related protein 5 gene (LRP5) underlie osteoporosis-pseudoglioma syndrome. Animal models implicate a role for LRP5 in lipid and glucose homeostasis. The objective was to evaluate metabolic consequences of LRP5 mutations in humans. DESIGN AND PATIENTS: Thirteen Finnish individuals with homozygous or heterozygous LRP5 mutations were assessed for bone health, glucose and lipid metabolism, and for serum serotonin concentration. Results were compared with findings in family members without mutations. MEASUREMENTS: Bone mineral density (BMD), vertebral morphology, oral and intravenous glucose tolerance tests, lipid profile and serum serotonin concentrations. RESULTS: Two individuals were homozygous for R570W, one compound heterozygous for R570W and R1036Q, and 10 were heterozygous (six for R570W, three for R1036Q and one for R925C). Subjects with two LRP5 mutations had multiple spinal fractures and low BMD. Subjects with one mutation had significantly lower median lumbar spine (P = 0.004) and femoral neck (P = 0.005) BMD Z-scores, and more often vertebral fractures than the 18 individuals without mutations. Of the 12 subjects with LRP5 mutation six had diabetes and one had impaired glucose tolerance. Intravenous glucose tolerance tests suggested impaired beta-cell function; no insulin resistance was observed. Prevalence of hypercholesterolaemia was similar in mutation positive and negative subjects. Serum serotonin concentrations showed a trend towards higher concentrations in subjects with LRP5 mutation. CONCLUSIONS: We found high prevalence of osteoporosis and abnormal glucose metabolism in subjects with LRP5 mutation(s). Further studies are needed to establish the role of LRP5 in glucose and lipid metabolism. PMID- 19673928 TI - A bioavailability/bioequivalence and pharmacokinetic study of two oral doses of torasemide (5 and 10 mg): prolonged-release versus the conventional formulation. AB - 1. The main objective of the present study was to compare the bioavailability/bioequivalence of a new prolonged-release (PR) formulation of torasemide with an immediate-release (IR) formulation. In addition, we assessed the pharmacokinetics of both formulations, as well as the urine pharmacodynamics. 2. Two doses (5 and 10 mg) of PR torasemide were compared with the same doses of IR torasemide in a single-blind, single-dose, two-treatment, two-period, cross over, sequence-randomized clinical trial in 20 healthy volunteers (two groups; n = 10 in each group). Torasemide plasma concentrations were measured by high pressure liquid chromatography-electrospray ionization mass spectrometry. Torasemide urine concentrations, the diuretic effect of torasemide, urine electrolytes and urine density were also determined. 3. Plasma bioequivalence parameters, based on logged values, were as follows: (i) in the 5 mg group, the area under the plasma drug concentration-time curve from t = 0 to last measurable drug concentration at time t (AUC(0-t)) tablet ratio was 1.03 (90% confidence interval (CI) 0.91-1.17) and C(max) was 0.82 (90% CI: 0.68-0.98); and (ii) in the 10 mg group, the AUC(0-t) was 1.07 (90% CI 0.99-1.14) and C(max) was 0.68 (90% CI 0.60-0.78). The PR formulation showed a significantly prolonged t(max) compared with the IR formulation. The amount of torasemide recovered in the urine 24 h after administration was higher with the PR formulation for both doses. The natriuretic rate versus torasemide excretion rate for the PR and IR formulations were successfully regressed to a sigmoid E(max) model. Pharmacodynamic urine evaluations were similar with both formulations, although urine volume and urine electrolyte excretion were lower for the PR formulation in the first hour after administration. However, the PR formulation showed higher natriuretic efficiency. No significant adverse events were reported. 4. In conclusion, both formulations of torasemide showed similar systemic exposure (AUC). However, the PR formulation had a lower rate of absorption (lower C(max) and prolonged t(max)). The PR formulation had urinary excretion rates that were associated with a higher natriuretic efficiency and more constant diuresis. PMID- 19673929 TI - Peroxisome proliferator-activated receptor gamma-independent effects of thiazolidinediones on human cardiac myofibroblast function. AB - 1. Thiazolidinediones (TZDs) are peroxisome proliferator-activated receptor (PPAR) gamma agonists that are used to lower insulin resistance in Type 2 diabetic patients. Although TZDs exhibit beneficial effects on the vasculature, their effects on the heart are less clear and are the subject of current clinical debate. Thiazolidinediones have been reported to reduce adverse myocardial remodelling, a pathology in which cardiac myofibroblasts (CMF) are pivotal. 2. The aim of the present study was to investigate whether TZDs modulate specific human CMF functions of importance to the myocardial remodelling process and to determine whether any of these effects were mediated via PPARgamma activation. 3. Immunoblotting of cultured human CMF homogenates revealed strong expression of PPARgamma (approximately 50 kDa). Three different TZDs (ciglitazone, rosiglitazone and troglitazone) and the endogenous PPARgamma ligand 15-deoxy delta(12,14)-prostaglandin J(2) (15d-PGJ(2)) inhibited CMF proliferation (cell number and expression of proliferating cell nuclear antigen) in a concentration dependent manner (range 0.1-10 micromol/L) with similar potencies. This antiproliferative effect of TZDs was not reversed by the PPARgamma antagonists GW9662 or T0070907 (10-25 micromol/L). None of the TZDs or 15d-PGJ(2) affected cell migration or invasion (Boyden chamber assays without or with Matrigel barrier), matrix metalloproteinase-2 or -9 secretion (gelatin zymography) or the actin cytoskeleton (rhodamine/phalloidin fluorescent confocal microscopy). 4. In conclusion, TZDs reduce human CMF proliferation via a PPARgamma-independent mechanism. Although TZDs do not inhibit CMF invasion, their antiproliferative activity may contribute to the ability of this class of drugs to modulate adverse myocardial remodelling. PMID- 19673930 TI - Erlotinib activates mitochondrial death pathways related to the production of reactive oxygen species in the human non-small cell lung cancer cell line A549. AB - 1. Erlotinib, a small-molecule epidermal growth factor receptor-tyrosine kinase (EGFR-TK) inhibitor, has been approved for the management of advanced non-small cell lung cancer. The aim of the present study was to investigate whether erlotinib exerts potent antitumour activities through the reactive oxygen species (ROS)-c-Jun N-terminal kinase (JNK) pathway in the human non-small cell lung cancer cell line A549. 2. The 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H tetrazolium bromide (MTT) assay and Hoechst 33342 staining showed that erlotinib produced a decline in cell viability of A549 cells and induced cell apoptosis, coupled with quick accumulation of ROS. In addition, erlotinib increased the respiratory control ratio, NADPH oxidase catalytic subunit gp91(phox) expression and superoxide anion (O2(-)) generation, suggesting that erlotinib induced ROS production in A549 cells from both mitochondrial and NADPH oxidase sources. 3. Fluorescence microscopy with the JC-1 probe and western blot analysis indicated that erlotinib induced loss of mitochondrial membrane potential, the release of cytochrome c and apoptosis-inducing factor (AIF) and activation of JNK. 4. The results of the present study suggest that erlotinib has potent antitumour activity in A549 cells by activating ROS-dependent, JNK-driven cell apoptosis. These findings provide a novel insight into the mechanism of action of erlotinib in the treatment of human non-small cell lung cancer in addition to its effects in inhibiting EGFR-TK. PMID- 19673931 TI - Behavioural changes induced by angiotensin-converting enzyme inhibition during pregnancy and lactation in adult offspring rats. AB - 1. The use of angiotensin-converting enzyme (ACE) inhibitors during pregnancy is contraindicated because of their association with increased risks of fetopathy, including central nervous systems malformations. In addition, some reports have shown that renin-angiotensin system components are expressed differently during embryonic development and adulthood in the rat. 2. Because angiotensin II and its derivative peptides have been implicated in anxiety and modulation of nociception, the aim of the present study was to investigate whether inhibiting ACE during prenatal and neonatal periods would alter behavioural plasticity in adult male offspring rats. 3. Female Wistar rats were treated with captopril (2 mg/mL water; approximately 200 mg/kg per day) during pregnancy and lactation. At adulthood, the offspring were subjected to the open field, elevated plus maze, social interaction, forced swimming and tail flick tests. 4. Perinatal captopril treatment significantly increased ambulation (33%; P < 0.05) and decreased resting time (37.5%; P < 0.05) in the open field test. Perinatal captopril treatment did not alter any of the behavioural parameters of the elevated plus maze; however, captopril treatment did cause a significant increase in social interaction (75.3%; P < 0.05). In the forced swimming test, there was an increased latency period (102.9%; P < 0.001) and a decreased immobility period (38.7, P < 0.05) in rats treated with perinatal captopril. In the tail flick test, perinatal captopril treatment significantly reduced the latency time (26.3%; P < 0.01). 5. The data show that ACE inhibition during prenatal and neonatal periods affects behavioural responses in adult offspring rats, suggesting that ACE is required for the development of neural systems that are associated with adult anxiety and nociceptive behavioural responses. PMID- 19673932 TI - Effect of calcium channel blockade on adrenergically induced renal vasoconstriction in rat models of renal impairment. AB - 1. Calcium channel blockade attenuates adrenergically induced renal vasoconstriction and the present study examined whether the magnitude of the supression was enhanced or blunted in rat models of renal failure or diabetes induced glomerular hyperfiltration. 2. Male Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR) were used that had either renal failure induced by cisplatin administration or early diabetic nephropathy (EDN) consequent to an injection of streptozotocin (STZ). After 7 days of cisplatin or 4 weeks of STZ, rats were anaesthetized and renal haemodynamic studies were performed. 3. Cisplatin-treated WKY rats and SHR exhibited reduced creatinine clearance (CCr) and increased fractional excretion of sodium (FE(Na)) and kidney index (all P < 0.05), along with tubular damage in the kidney, compared with non treated rats. In the EDN model, there was marked albuminuria and increased FE(Na), kidney index and CCr (all P < 0.05) compared with non-diabetic nephropathy (NDN) rats. Amlodipine significantly (P < 0.05) and dose-dependently attenuated the magnitude of the neurally and adrenergically induced renal vasoconstriction in all experimental groups compared with their respective control groups (13-6 vs 9-5% in renal failure vs non-renal failure WKY rats; 26 15 vs 17-8% in renal failure vs non-renal failure SHR; 19-9 vs 14-5% in EDN vs NDN). 4. The data obtained demonstrate that not only does amlodipine blunt adrenergically induced renal vasoconstriction in normal rats, but that it also does so, and to a greater extent, in rats with either renal failure or hyperfiltration. PMID- 19673933 TI - Activation of reactive oxygen species and the renin-angiotensin system in IgA nephropathy model mice. AB - 1. Although IgA nephropathy is the most common form of primary glomerulopathy, the detailed mechanisms underlying its development remain uncertain. 2. In the present study, we used male high IgA strain of ddY (HIGA) mice as the IgA nephropathy model and age-matched male BALB/c mice as the control. Recent studies have demonstrated that reactive oxygen species (ROS)-dependent enhancement of the renin-angiotensin system (RAS) plays a potential role in the development and progression of renal injury. Therefore, in the present study we periodically measured the systolic blood pressure (SBP) of mice over the period 21-25 weeks of age and estimated markers for ROS, RAS and renal damage after mice had been killed at 25 weeks of age. 3. Markers for ROS (urinary 8-isoprostane excretion and renal 4-hydroxy-2-nonenal accumulation), RAS (renal angiotensinogen protein expression, urinary angiotensinogen excretion and renal angiotensin II) and renal damage (desmin-positive area and urinary protein excretion), as well as SBP, were significantly increased in HIGA mice compared with control BALB/c mice. 4. The data suggest that both ROS and the RAS are activated at an early phase in IgA nephropathy model mice. PMID- 19673934 TI - GABA(A) and GABA(B) receptor-mediated inhibition of sympathetic outflow in the paraventricular nucleus is blunted in chronic heart failure. AB - 1. Alterations in the paraventricular nucleus (PVN) are reported to be involved in sympathetic overactivity in chronic heart failure (CHF). Inhibitory inputs into the PVN contribute to sympathetic outflow. The aim of the present study was to comparatively determine the role of GABA mechanisms in the PVN in the tonic control of cardiovascular activity in anaesthetized sham and CHF rats. 2. The CHF model was induced by coronary artery ligation. Unilateral microinjection of the GABA(A) receptor agonist muscimol (0.1-0.8 nmol/200 nL) or the GABA(B) receptor agonist baclofen (0.25-2.0 nmol/200 nL) into the PVN produced similar, dose dependent reductions in arterial pressure (AP), heart rate (HR) and renal sympathetic nerve activity (RSNA). This response was significantly blunted in CHF rats. In contrast, microinjection of the GABA(A) receptor antagonist bicuculline (25-200 pmol/200 nL) or the GABA(B) receptor antagonist CGP35348 (0.25-2.0 nmol/200 nL) into the PVN caused larger, dose-dependent increases in AP, HR and RSNA in sham than in CHF rats. 3. Polymerase chain reaction data showed that mRNA expression levels of the GABA(A) receptor alpha(1)-subunit and of the GABA(B1(a)) and GABA(B1(b)) receptor subtypes in the PVN were significantly lower in CHF than in sham rats. 4. The results of the present study suggest that the tonic inhibition mediated by both GABA(A) and GABA(B) receptors in the PVN on sympathetic outflow is blunted in CHF, which may be an important mechanism responsible for sympathetic hyperactivity in CHF. PMID- 19673935 TI - Histopathological and scintigraphic comparisons of the protective effects of L carnitine and amifostine against radiation-induced late renal toxicity in rats. AB - 1. The aim of the present study was to compare the protective effects of L carnitine and amifostine against radiation-induced late nephrotoxicity using technetium-99m diethylenetriaminepentaacetic acid scintigraphy and histopathological examination. 2. Seventy-one Albino rats were randomly divided into six groups as follows: (i) AMI + RAD (n = 15), 200 mg/kg, i.p., amifostine 30 min prior to irradiation (a single dose of 9 Gy); (ii) LC + RAD (n = 15), 300 mg/kg, i.p., L-carnitine 30 min prior to irradiation; (iii) LC (n = 10), 300 mg/kg, i.p., L-carnitine 30 min prior to sham irradiation; (iv) AMI (n = 10), 200 mg/kg, i.p., amifostine 30 min prior to sham irradiation; RAD (n = 11), 1 mL/kg, i.p., normal saline 30 min prior to irradiation; and (vi) control (n = 10), 1 mL/kg, i.p., normal saline 30 min prior to sham irradiation. Scintigraphy was performed before treatment and again 6 months after treatment. Kidneys were examined by light microscopy and a histopathological scoring system was used to assess the degree of renal damage. 3. The main histopathological findings were proximal tubular damage and interstitial fibrosis. Glomerular injury was similar in all groups. Tubular degeneration and atrophy were less common in the AMI + RAD group than in the RAD group (P = 0.011 and P = 0.015, respectively), as well as in the LC + RAD group compared with the RAD group (P = 0.028 and P = 0.036, respectively). Interstitial fibrosis in the AMI + RAD and LC + RAD groups was significantly less than that in the RAD group (P = 0.015 and P = 0.015, respectively). The highest total renal injury score (9) was seen in the RAD group. On scintigraphy, there were significant differences in post-treatment time to peak count (T(max)) and time from peak count to half count (T((1/2))) values (P = 0.01 and 0.02, respectively) between groups in the right kidney. In the control and RAD groups, the T((1/2)) of the right kidney was 8 +/- 2 and 21 +/- 2 min, respectively. The T(max) values for the AMI + RAD and LC + RAD groups (2.8 +/- 0.2 and 3.2 +/- 0.2 min, respectively) were similar to those in the control group (2.5 +/- 0.3 min). 4. Based on the results of the present study, L carnitine and amifostine have comparable and significant protective effects against radiation-induced late nephrotoxicity. PMID- 19673936 TI - Salivation triggered by pilocarpine involves aquaporin-5 in normal rats but not in irradiated rats. AB - 1. Using rats, we examined the muscarinic receptor subtype mediating pilocarpine induced parotid salivary secretion and the contributions of ion transporter systems (effluxes of K+ and Cl(-)) and aquaporin-5 (AQP5) translocation to this response in parotid glands in irradiated-induced xerostomia. 2. Salivary secretion was significantly lower in irradiated compared with sham-irradiated (normal) rats. In xerostomia rats, 0.4 and 0.8 mg/kg pilocarpine significantly increased parotid salivary secretion, although the salivary volume was still significantly less than in normal rats after the same dose of pilocarpine. 3. Pirenzepine (1 x 10(-6) to 1 x 10(-1) mol/L), AF-DX 116 (3 x 10(-6) to 3 x 10(-2) mol/L) and N-2-chloroethyl-4-piperidinyl diphenylacetate (4-DAMP; 1 x 10(-8) to 1 x 10(-2) mol/L) dose-dependently displaced radioligand binding to M(1), M(2) and M(3) receptors, respectively, in parotid membranes from both normal and irradiated rats. In each group of rats, 4-DAMP had the highest binding affinity. Pretreatment with 4-DAMP or pirenzepine dose-dependently inhibited pilocarpine induced parotid secretion in both normal and irradiated rats, with 4-DAMP being markedly more potent than pirenzepine. 4. Normal and irradiated-rat parotid cells did not differ significantly in terms of pilocarpine-induced changes in [Ca2+](i), [K+](i) and [Cl(-)](i). Pilocarpine markedly increased the amount of AQP5 in the apical plasma membrane of parotid cells isolated from normal but not irradiated rats. 5. Thus, pilocarpine induces parotid salivary secretion mainly via the M(3) receptor subtype in both irradiated and normal rats. The reduction in this pilocarpine-induced secretion seen in irradiated rats is due not to disturbances of intracellular Ca2+ mobilization or ion transporter systems, but rather to a disturbance of AQP5 translocation, which may be involved in the pathogenesis of X-ray irradiation-induced xerostomia. PMID- 19673937 TI - Intracerebroventricular injection of epidermal growth factor reduces neurological deficit and infarct volume and enhances nestin expression following focal cerebral infarction in adult hypertensive rats. AB - 1. Studies have documented the proliferative effects of epidermal growth factor (EGF) on neural progenitor cells in the normal or injured brain. The effect of EGF on post-stroke cerebral expression of nestin, a marker of neural progenitor cells, has not been examined in hypertensive rats. 2. In the present study, adult renovascular hypertensive Sprague-Dawley rats underwent either real or sham middle cerebral artery occlusion (MCAO). Intracerebroventricular injections of either 1 microg EGF or vehicle (0.01 mol/L phosphate-buffered saline containing 0.1 mg/mL rat serum albumin) were made 24 and 48 h after MCAO. Then, 1, 2, 3 and 4 weeks after MCAO, the postural reflex was evaluated in a blinded fashion before rat brains were processed to determine the infarct volume plus immunoreactivity for nestin and/or glial fibrillary acidic protein (GFAP). Another group of rats was used to quantify nestin expression using western blot analysis. 3. Middle cerebral artery occlusion resulted in a focal infarct that was largest at 1 week and diminished gradually over the time. The impaired postural reflex followed a similar time-course. In addition, MCAO induced a marked increase in nestin expression in both hemispheres, with a higher expression in the right hemisphere; this change was maximal at 1 week and largely subsided at 3 or 4 weeks. Within the right hemisphere, nestin expression was most pronounced in the subventricular and peri-infarct zones. Most nestin-immunoreactive cells were also positive for GFAP. 4. Thus, EGF treatment significantly increases nestin expression, reduces infarct volume and ameliorates postural reflex impairment in adult hypertensive rats. PMID- 19673939 TI - Efficacy and safety of atorvastatin during early hospitalization in elderly patients with unstable angina. AB - 1. Previous studies have demonstrated that early statin therapy after acute coronary syndrome decreases inflammation and mortality rates. The dose-response relationship for atorvastatin in elderly patients with unstable angina (UA) during early hospitalization in terms of lowering inflammatory factors, improving vascular endothelium function and safety is unclear. 2. In the present study, 166 consecutive patients with UA who were >/= 60 years of age were randomly assigned, in a double-blind manner, to receive 80 or 20 mg/day atorvastatin. High sensitivity C-reactive protein, interleukin-6, tumour necrosis factor-alpha, fibrinogen and lipid levels were measured at admission and 1, 2 and 8 weeks later. Vascular endothelial function was measured and the safety of the drug was monitored. 3. Levels of inflammatory factors were significantly lower in patients on 80 mg atorvastatin than in those on 20 mg atorvastatin at 2 and 8 weeks. Atorvastatin 80 mg not only resulted in a significant improvement in vascular endothelial function during early hospitalization for UA over that seen in patients on 20 mg atorvastatin, but also reduced lipid levels to a greater extent. At 8 weeks, almost all patients showed good tolerance of 80 mg/day atorvastatin. 4. The results of the present study indicate that intensive statin therapy with high-dose (80 mg/day) atorvastatin is more efficacious than and as safe as 20 mg/day atorvastatin when administered to elderly patients during early hospitalization for UA. PMID- 19673938 TI - Genetic models provide unique insight into angiotensin and bradykinin peptides in the extravascular compartment of the heart in vivo. AB - 1. There is continuing uncertainty about the tissue compartments where angiotensin and bradykinin peptide formation occurs. Mice with angiotensin converting enzyme (ACE) expression targeted to the cardiomyocyte membrane provide a unique experimental model to detect ACE substrates in the extravascular compartment of the heart in vivo. 2. Angiotensin (Ang) I and II, bradykinin-(1-7) and bradykinin-(1-9) were measured in blood and cardiac ventricles of wild-type (WT) mice, mice with a non-functional somatic ACE gene promoter (KO), mice homozygous (8/8) and heterozygous (1/8) for cardiomyocyte-targeted ACE expression and a non-functional somatic ACE gene promoter, and mice heterozygous for cardiomyocyte-targeted ACE expression and heterozygous for the WT ACE allele (WT/8). 3. Cardiac AngII levels of 8/8, 1/8, WT/8 and WT mice were higher than KO levels. Cardiac AngII levels in 8/8 and 1/8 mice were also higher than WT levels, but the levels in WT/8 mice were similar to WT levels. Cardiac bradykinin-(1-9) levels of WT, but not 8/8 mice, were lower than in KO mice, whereas bradykinin-(1 7) levels in 8/8 mice were lower than in KO mice. 4. We conclude that AngI and bradykinin-(1-7) are present in the cardiac extravascular compartment of mice lacking vascular ACE and that extravascular ACE produces AngII and metabolises bradykinin-(1-7) in this compartment. The data suggest that the vascular compartment is the main site of AngI and bradykinin-(1-9) formation and metabolism and that vascular ACE may limit AngI entry to the extravascular compartment of WT mice. PMID- 19673940 TI - Mechanical stretch induces endothelial nitric oxide synthase gene expression in neonatal rat cardiomyocytes. AB - 1. Mechanical stretch leads to cardiac hypertrophy and may ultimately cause heart failure. However, the effect of mechanical stretch on gene induction in cardiomyocytes remains to be determined. 2. In the present study, we compared transcript profiles of mechanically stretched neonatal rat cardiomyocytes with those of unstretched cells using cDNA microarrays. The microarrays contained probes for 480 known genes, including those involved in signal transduction, cell cycle regulation, the cytoskeleton and cell motility. Eighteen genes, including the eNOS gene, were identified as having significantly differential expression in response to mechanical stretch in cardiomyocytes. 3. Northern and western blot analysis further quantified the expression of the eNOS gene. Mechanical stretch increased constitutive NOS activity and nitric oxide (NO) production. The NO donor s-nitroso-N-acetylpenicillamine (SNAP) inhibited mechanical stretch stimulated protein synthesis, as measured by [3H]-leucine uptake. In addition, cardiomyocytes were infected with adenoviral vectors encoding cDNA for eNOS (Ad eNOS) and a phosphoglycerate kinase (PGK) empty vector (Ad-PGK). In contrast with Ad-PGK-infected cells, in cardiomyocytes infected with Ad-eNOS, there was increased calcium-dependent NOS activity and nitrite production. Cardiomyocytes infected with Ad-eNOS exhibited diminished mechanical stretch-stimulated protein synthesis. In contrast, in eNOS-knockdown cells, the increased eNOS protein levels and NOS activity induced by mechanical stretch were abolished, but protein synthesis was enhanced. 4. The results of the present study indicate that eNOS gene expression is induced by mechanical stretch, leading to increased constitutive NOS activity and NO production, which may be a negative regulator in cardiomyocyte hypertrophy. PMID- 19673941 TI - Effect of R219K polymorphism of the ABCA1 gene on the lipid-lowering effect of pravastatin in Chinese patients with coronary heart disease. AB - 1. In the present study, we investigated the effects of the R219K polymorphism of the ATP-binding cassette transporter A1 (ABCA1) gene on serum lipid levels and the response to statin therapy in Chinese patients with coronary heart disease (CHD). 2. The study population consisted of 365 patients with CHD and 246 control subjects without signs or symptoms of CHD. Patients with CHD were treated with 20 mg/day pravastatin. Fasting serum lipids were determined before and after 12 weeks of treatment. Genotyping was performed by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). 3. The R219K polymorphism of the ABCA1 gene was not significantly associated with CHD (P > 0.05). Compared with controls, patients with the RR genotype had significantly higher serum triglyceride levels and lower high-density lipoprotein-cholesterol (HDL-C) levels than those with the KK genotype (P < 0.05). In addition, the effects of pravastatin in increasing HDL-C levels were significantly greater in patients with the KK genotype compared with those with the RR genotype (P < 0.05). 4. In conclusion, the R219K polymorphism of ABCA1 was associated with altered lipoprotein levels and the R219K variant significantly modulated the HDL-C response to pravastatin in Chinese patients with CHD. PMID- 19673942 TI - Cellular expression of renal, cardiac and pulmonary inducible nitric oxide synthase in double-transgenic mice expressing human renin and angiotensinogen genes. AB - 1. Hypertensive mice expressing the human renin (REN) and angiotensinogen (AGT) genes are used as a model for human hypertension. 2. The aim of the present study was to investigate the cellular expression and distribution of inducible nitric oxide synthase (iNOS) using immunohistochemistry in lung, heart and kidney tissues from a model of human hypertension using male and female double transgenic (h-Ang 204/1h-Ren6) mice and wild-type C57/BI6J mice as controls. 3. In the kidney, the pattern of iNOS expression in various renal microanatomical regions during hypertension was similar to that of age-matched controls, except in the medullary ascending limb (MAL). In hypertension, iNOS expression was downregulated in the MAL. No significant differences in iNOS expression were seen between control or hypertensive mice in various cardiac microanatomical locations. In the lungs of hypertensive mice, iNOS expression was upregulated in bronchial airway epithelium and bronchial and vascular smooth muscle cells, but downregulated in alveolar macrophages, alveolar septa and pulmonary vascular endothelial cells. Expression of iNOS was similar between male and female mice in the kidney, heart and lungs. 4. In conclusion, iNOS regulation in hypertension is complex and depends on the cell type in which it is expressed and the localization of the cell type in the cardiorenal and pulmonary systems. PMID- 19673943 TI - Genetic variants of beta(1)-adrenoceptor gene polymorphisms (Ser49Gly and Arg389Gly) and essential hypertension in a south Indian Tamil population. AB - 1. Essential hypertension is a complex polygenic disorder, the pathogenesis of which is dependent on an interplay between genetic and environmental factors. Various studies suggest an association between beta(1)-adrenoceptor gene polymorphisms (Ser49Gly and Arg389Gly) and cardiovascular disorders, including hypertension, cardiomyopathy and congestive heart failure. 2. The genetic profile of the beta(1)-adrenoceptor gene has not yet been documented for any Indian population. Thus, the aim of the present study was to investigate the association between beta(1)-adrenoceptor gene polymorphisms and essential hypertension in a south Indian Tamil population. 3. The present case-control study included 438 patients with essential hypertensives and 444 healthy volunteers from the Tamil population. Genotyping was performed using real-time polymerase chain reaction. 4. Genotype and allele frequencies of Ser49Gly and Arg389Gly polymorphism were compared between hypertensive patients and healthy volunteers. The homozygous variant genotype Gly49Gly of the Ser49Gly polymorphism was higher in hypertensive patients compared with controls (12.3 vs 7.4%, respectively). After adjusting for confounding variables (odds ratio (OR) 2.0; 95% confidence interval (CI) 1.2-2.9; P < 0.01) by multilogistic regression analysis, the gene was found to be associated with hypertension. A significant interaction was observed in hypertensive patients carrying the Ser49Gly/Gly49Gly x Arg389Gly/Gly389Gly genotypes (OR 1.9; 95% CI 1.1 2.7). 5. In conclusion, the Ser49Gly polymorphism is associated with essential hypertension in a south Indian Tamil population. The results of the present study deviate from those of previous studies, implying that marked interethnic difference exist in beta(1)-adrenoceptor gene polymorphisms. PMID- 19673944 TI - Changes in the composition of the thoracic aortic wall in spontaneously hypertensive rats treated with losartan or spironolactone. AB - 1. In the present study, we compared the elastin and collagen content of thoracic aortic medial and adventitial layers from Wistar-kyoto (WKY) and spontaneously hypertensive rats (SHR). In addition, the effects of losartan, an angiotensin II receptor antagonist, and spironolactone, a mineralocorticoid receptor antagonist, on collagen and elastin content were determined. 2. Prehypertensive (4-week-old) and hypertensive (16-week-old) SHR were randomly divided into three groups treated with either 0.9% NaCl, losartan (20 mg/kg per day) or spironolactone (200 mg/kg per day). Prehypertensive and hypertensive SHR were treated for 12 and 16 weeks, respectively. Age-matched WKY rats were not treated with NaCl, losartan or spironolactone and served as the control group. 3. The medial and adventitial layers of the thoracic aorta were composed mainly of elastin and collagen, respectively, in both SHR and WKY rats. Compared with WKY rats, SHR exhibited greater collagen and elastin content in the media, but decreased collagen and elastin content in the adventitial layer. Both medial and adventitial collagen and elastin content increased significantly with age in both strains and was greater in 32-week-old rats compared with 16-week-old rats. Spironolactone treatment decreased collagen content in the media of thoracic aortas from prehypertensive SHR, whereas losartan decreased collagen content in the media of aortas from hypertensive SHR. In contrast, neither spironolactone nor losartan had any effect on adventitial collagen content in prehypertensive and hypertensive SHR. Medial collagen and elastin were positively related to pulse pressure (PP), but there was no correlation between adventitial mass or collagen content and PP or mean arterial pressure in untreated and treated SHR and WKY rats. 4. In conclusion, the composition of the medial and adventitial layers of the thoracic aorta differs and treatment of SHR with losartan and spironolactone decreases collagen content when delivered at the hypertensive or prehypertensive stage, respectively. However, neither drug has any effect on adventitial collagen content in SHR. PMID- 19673945 TI - The novel selective toll-like receptor 4 signal transduction inhibitor tak-242 prevents endotoxaemia in conscious Guinea-pigs. AB - 1. TAK-242 is a novel compound that suppresses nitric oxide and cytokine production by selectively inhibiting intracellular signals from toll-like receptor (TLR)-4. In the present study, we investigated the effectiveness of TAK 242 against sepsis using an endotoxaemia model in conscious and unrestricted guinea-pigs. Measures examined included muscle tension paralysis of the intestine, blood pressure, high morbidity group box (HMGB)-1 levels and survival rate. 2. Tension of the longitudinal muscle of the colon was monitored continuously by telemetry. Arterial blood pressure was monitored via a carotid artery catheter. TAK-242 was administered intravenously through a jugular vein catheter. Guinea-pigs were divided into a control group, given vehicle (placebo emulsion), and the experimental group, administered 3 or 10 mg/kg TAK-242, 1 h before administration of 10 mg/kg lipopolysaccharide (LPS). 3. In the control group, the tension of the longitudinal muscle of the colon decreased in a time dependent manner and blood pressure was reduced, with maximal effects observed 1 3 h after administration of LPS. In the TAK-242-treated group, LPS-induced relaxation of the intestine and hypotension were significantly inhibited. In the control group, HMGB-1 levels were increased after LPS administration and this reaction was significantly blocked in the TAK-242-treated group. Importantly, survival rate was increased after TAK-242 treatment. 4. In conlusion, the results of the present study show that TAK-242 inhibited the symptoms associated with endotoxaemia in a guinea-pig model of sepsis and that it may, therefore, be an effective treatment for sepsis. PMID- 19673946 TI - Hyperbaric oxygenation applied immediately after coronary occlusion reduces myocardial necrosis and acute mortality in rats. AB - 1. Because in ischaemia there is a critical lack of O2, it has been reasoned that increasing O2 delivery to the ischaemic myocardium could serve as adjunctive therapy for acute myocardial infarction (MI). Accordingly, in the present study, the effect of early hyperbaric oxygenation (HBO) on mortality and MI size after coronary occlusion was examined in rats. 2. After coronary occlusion, male Wistar rats were randomly assigned to receive either HBO for 1 h in a hyperbaric chamber (100% O(2) at 253 kPa; n = 106) or ambient O2 as the control (n = 111). The extent of myocardial necrosis was assessed (triphenyltetrazolium) immediately after treatment in the HBO (n = 50) and control (n = 47) groups. The remaining rats were evaluated 24 h after occlusion to enable calculation of MI size and mortality. 3. Immediately after therapy, the size of the MI was significantly greater in the control group compared with that in the HBO group (40 +/- 3 vs 27 +/- 2% of the left ventricle (LV), respectively; P < 0.001). The 24 h mortality of control rats was higher than that of HBO rats (34 vs 16%, respectively; P = 0.02). Control rats that survived 24 h had a larger MI than did HBO rats that survived 24 h (40 +/- 4 vs 29 +/- 3% of the LV, respectively; P = 0.005). Furthermore, large necrotic areas (> 40% of the LV) were more frequent in control than HBO rats (55 vs 27% of infarcted hearts, respectively; P = 0.01). There was less pulmonary congestion observed in HBO rats compared with control rats. 4. In conclusion, early therapy with HBO during the onset of an acute ischaemic event decreases the necrotic area and reduces acute mortality. These data support further investigation of HBO as an adjuvant therapy for acute MI. PMID- 19673947 TI - SYNGAP: bridging the gap between genetic factors and autosomal non-syndromic mental retardation. PMID- 19673948 TI - IFRD1 modulates disease severity in cystic fibrosis through the regulation of neutrophil effector function. PMID- 19673949 TI - MCAD mutations identified in newborn screening cause different levels of enzymatic dysfunction. PMID- 19673950 TI - Novel mutations of the CLCN5 gene including a complex allele and A 5' UTR mutation in Dent disease 1. PMID- 19673951 TI - Parental SCN1A mutation mosaicism in familial Dravet syndrome. AB - Different SCN1A mutations are known to cause a variety of phenotypes, such as generalized epilepsy with febrile seizures plus (GEFS+), Dravet syndrome and familial hemiplegic migraine (FHM). In Dravet syndrome, most mutations are de novo and familial cases are rare. In this study, Dravet syndrome is observed in two maternal half sisters. They have healthy fathers and their common mother has never experienced seizures, but has a lifelong history of migraine. Direct sequencing of DNA extracted from blood revealed a heterozygous SCN1A nonsense mutation c.3985C>T in the sisters, but not in the mother. The mutation induces a premature stop codon and probably leads to a non-functional protein. Further examination of the mother's DNA showed that she has a mosaicism of the mutation. This report of parental SCN1A nonsense mutation mosaicism in familial Dravet syndrome suggests that mosaicism might be more common than previously suspected and emphasizes the importance of taking mosaicism into account in genetic counselling of Dravet syndrome and SCN1A mutations. Furthermore, whether the migraine of the mother could be influenced by her SCN1A mutation mosaicism is not known, but increased awareness of migraine in future studies of SCN1A related epilepsies could clarify this intriguing link between migraine and epilepsy. PMID- 19673952 TI - Phenotypic and molecular characterization of a novel DCX deletion and a review of the literature. PMID- 19673953 TI - Usefulness of the aesthetic result as a success criterion for implant therapy: a review. AB - PURPOSE: The purpose of this study was to review the reported evaluation criteria of the aesthetic result in oral implant rehabilitation. MATERIALS AND METHODS: A literature search of MEDLINE, the Cochrane Collaboration, and EMBASE was performed to retrieve studies published between January 1990 and December 2008 using the following key words: "dental implants,""clinical trial," and "aesthetic index" (and their synonyms). A manual search of the literature published in the same period was also carried out using the following publications: Clinical Oral Implant Research, The International Journal of Oral and Maxillofacial Implants, and The International Journal of Periodontics and Restorative Dentistry. The inclusion criteria of the published studies were the following: human clinical trial, oral implant rehabilitation, at least 10 implants, at least 6 months of follow-up from insertion of the prosthesis, and evaluation of the aesthetic result by means of an index. RESULTS: The literature search revealed 650 relevant bibliographic references, of which 89 were selected for further analysis. A final total of 29 articles fulfilled the inclusion criteria; these included 10 retrospective case series, 11 prospective case series, 1 retrospective controlled clinical trial, 1 prospective controlled clinical trial, and 6 randomized controlled clinical trials. In general, evaluations of aesthetic results appear only in the more recent studies and refer mostly to implant rehabilitation in the maxillary anterior zone; the index used, in most cases, was the Papilla Index of Jemt. CONCLUSIONS: Although there appears to be a growing interest in aesthetics in dental implantology, there are as yet no universally accepted evaluation criteria of the aesthetic result. Therefore, further research is necessary to establish a common, complete, and reproducible index for the evaluation of aesthetic outcome that can add in the success criteria for implant therapy in the maxillary and mandibular anterior areas. PMID- 19673954 TI - A 10-year prospective study of single tooth implants placed in the anterior maxilla. AB - PURPOSE: The aim of the study was to evaluate the biological and technical outcomes of early and delayed placed single tooth implants after 10 years of follow-up. MATERIALS AND METHODS: Twenty consecutive patients who needed a single tooth replacement in the anterior maxilla were included in this study. Ten implants were placed with an early placement protocol, that is, 4 weeks after tooth extraction, whereas the other 10 implants were placed with a conventional delayed placement protocol, for example, 12 weeks after tooth extraction. At the baseline and at the annual re-examinations, pain from implant regions, mobility, plaque, mucositis, and marginal bone levels were recorded. Biological and technical complications were registered. The patients evaluated the functional and aesthetic outcomes subjectively on a visual analogue scale after 3 and 10 years. RESULTS: All implants were still in situ after 10 years. The cumulative implant survival rate was 100%. Two implant-supported crowns were remade because of ceramic fractures. The 10-year crown survival rate was 90%. No significant differences in implant survival rates were found between the early and delayed protocols, regarding plaque retention, mucositis, or marginal bone levels. After 10 years, the mean marginal bone loss at both sides of the implants was less than 1.0 mm in the two groups. During the 10-year interval, 1 patient lost more than 1.5 mm of marginal bone, 3 patients lost between 1.0 mm and 1.4 mm of bone, and 16 patients lost less than 1.0 mm of bone as an average of the mesial and distal bone loss. CONCLUSIONS: This 10-year prospective clinical trial demonstrated a 100% implant survival and a 90% crown survival. The average marginal bone level change was less than 1 mm, and there was no difference between early and delayed implant placement. Patient satisfaction with the implant-supported single crowns reduced with time. PMID- 19673955 TI - Thiel embalming technique: a valuable method for teaching oral surgery and implantology. AB - BACKGROUND: Because of its high requirements on surgical experience and the need of complete understanding of the anatomy, oral surgery and especially implantology belong to the most demanding procedures in dentistry. Therefore, hands-on courses for oral surgery and implantology are considered a prerequisite to prepare for clinical practice. To achieve teaching conditions as realistic as possible, we used a novel human cadaver embalming method enabling tissue dissection comparable with the living body. METHODS: Thirty cadavers which were offered by the Institute of Anatomy for the purpose of running oral surgery and implantology courses were embalmed in the technique described by Thiel. On each cadaver, dissection of soft and hard tissue and implantological procedures were performed according to a structured protocol by each course participant. The conservation of fine anatomical structures and the suitability of the embalmed tissue for dissecting, drilling, and suturing were observed and photographically documented. RESULTS: By means of the Thiel embalming technique, oral surgery and implantological procedures could be performed under realistic conditions similar to the living body. Due to the conservation procedure, preparations could be carried out without any time limit, always maintaining the same high quality of the tissue. The maxillary sinus membrane, mucosa, bone, and nerves could be exposed and allowed dissecting, drilling, and suturing even after weeks like fresh specimens. CONCLUSION: The Thiel embalming method is a unique technique which is ideally suited to practice and teach oral surgery and implantology on human material. PMID- 19673956 TI - Flapless implant surgery using a mini-incision. AB - BACKGROUND: Traditional flapless implant surgery using a soft tissue punch device requires a circumferential excision of keratinized tissue at the implant site. A new flapless implant technique that can submerge implant fixtures is needed. PURPOSE: This article describes a flapless implant surgery method using a mini incision and compares the effects of soft tissue punch and mini-incision surgery on both the amount of osseointegration and the bone height around the implants using a canine mandible model. MATERIALS AND METHODS: Bilateral, edentulated, flat alveolar ridges were created in the mandibles of six mongrel dogs. After a 3 month healing period, two implants were placed on each side of the mandible using either soft tissue punch or mini-incision procedures. After an additional 3-month healing period, a second stage surgery and transmucosal abutment attachment was performed for mini-incision implant cases. Following a 2-month healing period, the dogs were sacrificed to evaluate the osseointegration and bone height around the implants. RESULTS: Average bone height was 9.6 +/- 0.4 mm in the soft tissue punch group and 9.8 +/- 0.3 mm in the mini-incision group (p > .05). Average osseointegration was 70.4 +/- 6.3% in the soft tissue punch group and 71.2 +/- 7.1% in the mini-incision group (p > .05). No significant differences were noted between the two groups in vertical alveolar ridge height or bone/implant contact. CONCLUSIONS: Our findings support the clinical use of mini-incision implant surgery at sites where implants need to be protected below the soft tissue during the early phase of healing, particularly for patients with poor bone quality and/or low primary implant stability. PMID- 19673957 TI - Mandibular incisive canal: cone beam computed tomography. AB - PURPOSE: Panoramic radiography is often used to analyze the anatomical structure of the teeth, jaws, and temporomandibular joints. Cone beam computed tomography (CBCT) imaging allows multiple axial slices of the image to be obtained through these anatomical structures. The aim of this study was to assess CBCT compared with panoramic radiography to verify the presence, location, and dimensions of the mandibular incisive canal. MATERIALS AND METHODS: CBCT scan images and panoramic radiographs of 89 subjects were compared for the presence of the mandibular incisive canal, its location, size, and anterior-posterior length. The distance between the incisive canal and the buccal and lingual plate of the alveolar bone, and the distance from the canal to the inferior border of the mandible and the tooth apex were also measured. A paired t-test was used to calculate any significant difference between the two imaging techniques. RESULTS: Eighty-three percent of the CBCT scans showed the presence of the incisive canal, as did 11% of the panoramic radiographs. The range of the incisive canal diameter, as seen in the CBCT scans, was from 0.4 * 0.4 mm to 4.6 * 3.2 mm. The mean length of the canal was 7 +/- 3.8 mm. The distance from the inferior border of the mandible to the canal was 10.2 +/- 2.4 mm, and the mean distance to the buccal plate was 2.4 mm. The apex-canal distance (in dentate subjects) was 5.3 mm. CONCLUSION: The presence, location, and dimensions of the mandibular incisive canal are better determined by CBCT imaging than by panoramic radiography. PMID- 19673958 TI - Combining scaffolds and osteogenic cells in regenerative bone surgery: a preliminary histological report in human maxillary sinus augmentation. AB - PURPOSE: The following case series evaluated the maxillary sinus augmentation responses to tissue-engineered bone graft obtained by a culture of autogenous osteoblasts seeded on polyglycolic-polylactic scaffolds and calcium phosphate. MATERIALS AND METHODS: Sinus floor augmentation was performed bilaterally in five patients (mean age 58.4 years) with tissue-engineered bone (test site - Oral Bone, BioTissue, Freiburg, Germany) or calcium phosphate (control site - Biocoral, Novaxa Spa, Milan, Italy). Biopsies were harvested 6 months after sinus augmentation for histometric evaluation. Volumetric measurements were taken at baseline and 6 months after the surgical procedure. RESULTS: The mean of vertical bone gain was 6.47 +/- 1.39 mm and 9.14 +/- 1.19 mm to test and control sites, respectively. The histological sections depicted mature bone with compact and cancellous areas. All biopsies contained varying percentages of newly formed bone and marrow spaces. The mean of bone tissue in the grafted area was 37.32 +/- 19.59% and 54.65 +/- 21.17% for tissue-engineered bone and calcium phosphate, respectively. CONCLUSION: Within the limits of the present report, the histological data in humans confirmed that tissue-engineered bone and calcium phosphate allowed newly formed bone after maxillary sinus augmentation. PMID- 19673959 TI - Evaluation of dimensional accuracy of panoramic cross-sectional tomography, its ability to identify the inferior alveolar canal, and its impact on estimation of appropriate implant dimensions in the mandibular posterior region. AB - OBJECTIVES: To evaluate the dimensional accuracy of panoramic cross-sectional tomography, its impact on implant size estimation and its ability in identifying the inferior alveolar canal in the mandibular posterior region. MATERIAL AND METHODS: Eight partially edentulous mandibles with 18 edentulous sites were obtained. Orthopantomograms and tomograms were made and the mandible's outline and the position of mandibular canals on tomograms were traced on a clear acetate paper. Horizontal and vertical magnification factors were calculated. The mandibular height, distance between mandibular canal and alveolar crest, maximum bucco-lingual width, distance between buccal cortex and mandibular canal, and cortical thickness at the inferior border of the mandible were measured. Potential implant sites were identified and implant sizes were estimated. Location and visibility of mandibular canals were also evaluated. The mandibles were sectioned at each site and all the above mentioned parameters were assessed which served as gold standard. RESULTS: Mean horizontal and vertical magnification factors were 1.47 +/- 0.048 and 1.53 +/- 0.038. Total height and maximum bucco-lingual width were underestimated by 1.88% and 1.59%. Crest to canal distance, cortical thickness at the inferior border of the mandible and buccal cortex to mandibular canal were overestimated by 0.59%, 5.16%, and 3.64%. Implant sizes were estimated for 11 sites and changes were recorded at 2 sites between record 1 and record 2. However, there was no disagreement between record 2 and record 3. Of the canals, 61.11% were located lingually and the visibility of mandibular canals was poor in 44.44% of cases. CONCLUSIONS: The tomograms were found to be accurate for the measurements in both horizontal and vertical planes and reliable for implant size estimation, taking into consideration proper magnification factors. They were also found to be useful in assessing the location of mandibular canal but were not very effective in discerning it. PMID- 19673960 TI - Temporary implants for patients with low bone density of the upper maxilla: an alternative rehabilitation strategy. AB - BACKGROUND: The use of removable provisional prosthesis after implant placement is undesirable for many patients. In addition, removable prostheses place unfavorable pressure over implants during the healing period. PURPOSE: The use of temporary implants enable greater stability and functionality of the prostheses. MATERIALS AND METHODS: This clinical report shows the use of temporary implants in a low density maxilla in conjunction with a fixed provisional prosthesis. RESULTS: The temporary implants contributed to the support of the provisional fixed upper arch prosthesis and to patient comfort with a fixed prosthesis during the healing period. CONCLUSIONS: Fixed provisional prostheses are a potentially desirable alternative treatment strategy. PMID- 19673961 TI - Aetiology of, and risk factors for, recurrent community-acquired pneumonia. AB - Recurrent community-acquired pneumonia (CAP) requiring hospitalization is a matter of particular concern. However, current information on its prevalence, aetiology and risk factors is lacking. To address these issues, we performed an observational analysis of a prospective cohort of hospitalized adults with CAP. Recurrence was defined as two or more episodes of CAP 1 month apart within 3 years. Patients with severe immunosuppression or local predisposing factors were excluded. Of the 1556 patients, 146 (9.4%) had recurrent CAP. The most frequent causative organism was Streptococcus pneumoniae, both in patients with recurrent CAP and in those without recurrence. Haemophilus influenzae, other Gram-negative bacilli and aspiration pneumonia were more frequent among patients with recurrent CAP, whereas Legionella pneumophila was rarely identified in this group. Independent factors associated with recurrent CAP were greater age, lack of pneumococcal vaccination, chronic obstructive pulmonary disease (COPD) and corticosteroid therapy. In a sub-analysis of 389 episodes of pneumococcal pneumonia, the only independent risk factor for recurrence was lack of pneumococcal vaccination. Recurrence of CAP is not a rare clinical problem and it occurs mainly in the elderly, patients with COPD, and those receiving corticosteroids. Our study provides support for recommending pneumococcal vaccination for adults at risk of pneumonia, including those with a first episode of CAP. PMID- 19673962 TI - Klebsiella pneumoniae bacteraemia in a region of Canada. AB - The second case of magA+ rmpA+ hypermucoviscosity phenotype Klebsiella pneumoniae infection was documented in Canada, in an immigrant from Algeria. To ascertain whether this represented recent importation of the strain or local transmission within Canada, a retrospective study of K. pneumoniae bacteraemia was conducted in the region, from 1997 to 2007, and 411 episodes were identified. No epidemiological evidence for local transmission of this strain was found. However, for the first time, the population incidence of K. pneumoniae bacteraemia was determined, which increased by 82% between 1997 and 2007, from 10.2 to 18.7 per 100 000 inhabitants. Incidence increased dramatically with age and with the presence of diabetes, but remained stable over time within each stratum. The proportion of patients with K. pneumoniae bacteraemia who were diabetic increased from 26% (1997-2004) to 42% (2005-2007). The rising incidence of K. pneumoniae bacteraemia may represent an unexpected consequence of the expanding population of adult diabetics. PMID- 19673963 TI - Dissemination and genetic context analysis of bla(VIM-6) among Pseudomonas aeruginosa isolates in Asian-Pacific Nations. AB - VIM-6, previously reported in two strains from Singapore recovered in 2000, was detected in 16 isolates collected in 2006 in India (12 isolates), Indonesia (two), Korea and the Philippines (one each). High genetic variability was observed among VIM-6-producing isolates (12 ribotypes and 11 pulsed-field gel electrophoresis types), but clones were observed in India and Indonesia; bla(VIM 6)-carrying integrons of 3.9 kb and 5 kb were detected, and two of five Indian hospitals yielded isolates with both integrons. These two integrons, bla(VIM-6) was located in the first position, followed by bla(OXA-10) and aacA4. The 5-kb integrons also harboured aadA1 and a 331-bp open reading frame encoding a putative efflux pump. PMID- 19673964 TI - Linking Pneumocystis jiroveci sulfamethoxazole resistance to the alleles of the DHPS gene using functional complementation in Saccharomyces cerevisiae. AB - Curative and prophylactic therapy for Pneumocystis jiroveci pneumonia relies mainly on cotrimoxazole, an association of trimethoprim and sulfamethoxazole (SMX). SMX inhibits the folic acid pathway through competition with para aminobenzoic acid (pABA), one of the two substrates of the dihydropteroate synthase (DHPS), a key enzyme in de novo folic acid synthesis. The most frequent non-synonymous single nucleotide polymorphisms (SNPs) in P. jiroveci DHPS are seen at positions 165 and 171, the combination leading to four possible different genetic alleles. A number of reports correlate prophylaxis failure and mutation in the P. jiroveci DHPS but, because of the impossibility of reliably cultivating P. jiroveci, the link between DHPS mutation(s) and SMX susceptibility is not definitively proven. To circumvent this limitation, the yeast Saccharomyces cerevisiae was used as a model. The introduction of the P. jiroveci DHPS gene, with or without point mutations, directly amplified from a clinical specimen and cloned in a centromeric plasmid into a DHPS-deleted yeast strain, allowed a fully effective complementation. However, in the presence of SMX at concentrations >250 mg/L, yeasts complemented with the double mutated allele showed a lower susceptibility compared with strains complemented with either a single mutated allele or wild-type alleles. These results confirm the need for prospective study of pneumocystosis, including systematic determination of the DHPS genotype, to clarify further the impact of mutations on clinical outcome. Additionally, the S. cerevisiae model proves to be useful for the study of still uninvestigated biological properties of P. jiroveci. PMID- 19673965 TI - A pyrosequencing method for molecular monitoring of regions in the inhA, ahpC and rpoB genes of Mycobacterium tuberculosis. AB - In this study, a pyrosequencing method for monitoring two genes related to isoniazid (INH)-resistance and a region of the rpoB gene linked to rifampin (RMP) resistance in Mycobacterium tuberculosis was developed and evaluated. Specifically, a 20-base pair (bp) region of inhA (from -24 to -4), a 35-bp region of ahpC (from -39 to -4), and a 57-bp region of rpoB (from codon 515 to 533) were analysed by pyrosequencing. For the development of the method, selected non consecutive clinical isolates of M. tuberculosis were analysed, including: 25 isolates susceptible to both INH and RMP, 18 RMP-monoresistant isolates, 17 INH monoresistant isolates, and 15 multidrug-resistant strains. Our pyrosequencing methodology was further evaluated using 96 M. tuberculosis isolates. Mutations in ahpC were found to be associated with INH resistance (p <0.05). By setting any mutation in ahpC as a marker of resistance, the specificity and the positive predictive value (PPV) were 100%. Similarly, any mutation in the rpoB gene was associated with a RMP resistance phenotype (p <0.01). Using any mutation in rpoB as a marker of RMP resistance, the sensitivity of this assay was 73% and the specificity and PPV were 100%. The use of this pyrosequencing method to analyse the ahpC and rpoB genes allowed us to detect INH- and/or RMP-resistant isolates. Furthermore, this method represents an opportunity to expedite the description of novel mutations related to drug resistance. PMID- 19673966 TI - Azole resistance in allergic bronchopulmonary aspergillosis and Aspergillus bronchitis. AB - Oral azole antifungal therapy is used extensively for all forms of aspergillosis, including allergic bronchopulmonary aspergillosis (ABPA). However, long-term therapy may increase the risk of resistance. Here we report itraconazole and voriconazole resistance with reduced susceptibility to posaconazole in Aspergillus fumigatus in two patients exposed to itraconazole. Patients were diagnosed with ABPA and Aspergillus bronchitis related to innate immune defects. An azole susceptible strain was initially isolated from patient 1, but later a genetically different azole-resistant strain was cultured, possibly related to sub-therapeutic itraconazole levels, which could be a trigger for selection of resistance. The mechanism of resistance identified in this case was an L98H change in Cyp51A, accompanied by a tandem repeat in the promoter region of cyp51A leading to increased expression. No cyp51A mutation was found in azole-resistant isolates recovered from patient 2. Both patients responded to posaconazole, with plasma levels of >1.0 mg/L. Subsequently, susceptible strains of different molecular types were cultured from both patients, suggesting eradication and replacement. PMID- 19673967 TI - Fatal Mediterranean spotted fever in Greece. AB - Forty-five days after the first confirmed and fatal Crimean-Congo haemorrhagic fever (CCHF) case in Greece in 2008, a female patient with similar signs and symptoms (high fever, thrombocytopaenia) and resident of the same area, was admitted to the University General Hospital of Alexandroupolis. Before admission, she had visited a local hospital where a cephalosporin was prescribed. A rash manifested over subsequent days, which was misdiagnosed as an allergy to the drug. Upon admission to the University Hospital, she was given further antibiotics, including doxycycline; a few hours later, ribavirin was added because CCHF was suspected. After the patient's death, rickettsiosis caused by Rickettsia conorii conorii (Meditteranean spotted fever; MSF) was diagnosed. Extremely high values of interleukin (IL)-1ra, IL-6, interferon-gamma-inducible protein-10, monocyte chemoattractant protein-1 and an absence of tumour necrosis factor-alpha were observed. MSF is a potentially severe and even fatal disease resembling viral haemorrhagic fevers that has to be included in the differential diagnosis of febrile syndromes combined with thrombocytopaenia, even when a tick bite is not reported, and an eschar is absent. Physicians have to be aware of MSF in patients with severe disease who are returning from the Mediterranean area. PMID- 19673968 TI - Changing epidemiology of invasive pneumococcal disease following increased coverage with the heptavalent conjugate vaccine in Navarre, Spain. AB - The present study evaluated changes in the incidence of invasive pneumococcal disease (IPD) and the pattern of serotypes isolated in Navarre, Spain, after the introduction and increased coverage of the heptavalent pneumococcal conjugate vaccine (PCV7). All cases with isolation of pneumococcus from normally sterile bodily fluids were included. The incidence of IPD in children and adults was compared for the periods 2001-2002 and 2006-2007. By the end of 2002, only 11% of children aged <5 years had received any dose of PCV7, whereas, beginning in 2007, the proportion exceeded 50%. Among the cases of IPD aged <5 years, the percentage of those vaccinated increased from 7% during 2001-2002 to 53% during 2006-2007 (p <0.001). The incidence of IPD from PCV7-serotypes decreased by 85% in children <5 years (p <0.001), by 45% in the population aged 5-64 years (p 0.10) and by 68% in those >or=65 years (p 0.004). By contrast, the incidence of IPD from non-PCV7 serotypes increased by 40% overall (p 0.006). The incidence of IPD from all serotypes did not change significantly in children <5 years (from 83 to 72 per 100 000) or in the total population (from 15.8 to 16.3 per 100 000). The percentage of cases as a result of serotypes 7 and 19A increased significantly in both children and adults. No significant changes were seen in the clinical forms of IPD. The pattern of serotypes causing IPD has changed, in both children and adults, following the increased coverage of PCV7, although the incidence has been reduced only slightly. PMID- 19673969 TI - Bacteraemia as a result of Campylobacter species: a population-based study of epidemiology and clinical risk factors. AB - Invasive disease as a result of Campylobacter is rarely reported. We reviewed 46 cases of blood stream infection with Campylobacter in a Danish population with complete follow-up. The incidence was 2.9 per 1 million person-years with a peak incidence in the age group above 80 years. In the population, the ratio of notified bacteraemia/enteritis patients with Campylobacter infection was 0.004. Patients with bacteraemia were older and had higher comorbidity, e.g. alcoholism, immunosuppression, previous gastrointestinal surgery or HIV infection. We found 26% of blood isolates resistant to ciprofloxacin. The length of hospitalization was significantly longer in bacteraemia patients, whereas the outcome was favourable with 28-day mortality of 4% in bacteraemia patients and 1% in enteritis patients. None of the bacteraemia patients relapsed within 365-day follow-up. PMID- 19673970 TI - Invasive fungal infections: past achievements and challenges ahead. PMID- 19673971 TI - Antifungal pharmacokinetics and pharmacodynamics: bridging from the bench to bedside. AB - This review considers a way in which experimental data can be used to identify safe and effective antifungal regimens for humans. The process begins with experimental models of invasive fungal infections that enable definition of optimal dosages and schedules of antifungal drug administration to be defined. These preclinical models also enable the identification of drug exposure targets that are associated with therapeutic outcomes of interest. Human pharmacokinetic variability results in a considerable range of drug exposures following the use of fixed antifungal drug regimens. This variability can be quantified using population pharmacokinetic modeling techniques. Monte Carlo simulation can then be used to simulate pharmacokinetic variability and thereby estimate the proportion of patients with a therapeutic outcome of interest. Effective and safe regimens can thus be studied appropriately in clinical settings. This approach can, and should, be used to optimize antifungal therapy for a large number of clinical scenarios. PMID- 19673972 TI - Invasive fungal infections in the paediatric and neonatal population: diagnostics and management issues. AB - Invasive fungal infections in children appear to have increased over the past few decades. Especially neonates and children with primary and secondary immunodeficiencies are at risk. Candida and Aspergillus spp. are the most commonly isolated organisms. In addition, Malassezia may cause systemic infections in newborns and zygomycosis is important because of its rising incidence and high case fatality rate. Timely diagnosis and initiation of appropriate antifungal therapy is imperative for improving outcomes. However, traditional techniques are time-consuming and representative sample material, using invasive procedures, may be difficult to obtain in the paediatric setting. This review provides an overview of the advances in detection and rapid species identification, with a focus on issues relevant in these settings. Subsequently, the current antifungal treatment options for neonates and children are discussed in light of the antifungal spectrum of the available agents and the specific pharmacokinetic properties in different age groups. Although a multitude of newer antifungal compounds have become available within the last decade, further studies are necessary to clearly establish the role for each of these agents among neonates and children. PMID- 19673973 TI - Trends in invasive fungal infections, with emphasis on invasive aspergillosis. AB - Patterns of invasive fungal infections are changing in many ways. Although yeast infections appear to have reached a stable incidence, the number of infections as a result of Aspergillus species appears to be increasing. Especially for mould infection, the diagnosis remains difficult and the detection and identification of clinically relevant isolates to the species level requires new validated techniques. Diagnostic tests are becoming more accurate, with biological markers such as PCR, galactomannan and 1,3 beta-D-glucan undergoing clinical validation. This is of importance because an early diagnosis is associated with increased survival. Correct diagnosis and in vitro susceptibility testing are becoming imperative for guidance of therapy in the context of changing epidemiology and the emergence of acquired resistance to antifungal drugs, as is insight into host factors that increase susceptibility to invasive mould infection and into the risks associated with new treatment modalities of underlying diseases. Despite improvements in the survival rates of patients with invasive fungal infection in recent years, continued research is required to meet the challenges associated with changes in epidemiology and resistance development. PMID- 19673976 TI - Surgery of the parotid gland: evolution of techniques, nomenclature and a revised classification system. PMID- 19673977 TI - Psychometric properties of the Chinese (Cantonese) Tinnitus Handicap Inventory. AB - OBJECTIVES: To establish the reliability and validity of the Chinese (Cantonese) version of the Tinnitus Handicap Inventory to measure the self-perceived handicapping effect and severity of the condition in patients with chronic tinnitus. DESIGN: Cross-sectional psychometric validation study. SETTING: Audiology clinics in a hospital setting. PARTICIPANTS: Subjects were 114 adult Chinese who attended the audiology clinics with a complaint of tinnitus. MAIN OUTCOME MEASURES: Test-retest and internal consistency reliability; construct validity. RESULTS: The Chinese version of the Tinnitus Handicap Inventory and its subscales showed good internal consistency reliabilities (alpha = 0.72-0.94) that are comparable to those of the original version. High correlations were observed between the Tinnitus Handicap Inventory and psychological distress, tinnitus related problem ratings and severity ratings. Factor analysis showed that the Chinese version of the Tinnitus Handicap Inventory has a unifactorial structure. A high degree of test-retest reliability was observed (intraclass correlation coefficient = 0.88). CONCLUSIONS: The results suggest that the Chinese (Cantonese) version of the Tinnitus Handicap Inventory is a reliable and valid measure of general tinnitus-related distress that can be used in clinical settings to quantify the impact of tinnitus on daily life. PMID- 19673978 TI - Asymmetric sensorineural hearing thresholds in the non-noise-exposed UK population: a retrospective analysis. AB - OBJECTIVES: To estimate the distribution of inter-aural sensorineural hearing threshold level differences in the non-noise-exposed adult population of the UK. SETTING: A two-stage population study carried out in 1979-1986, initially by postal questionnaire, followed up in a proportion of participants by clinical and audiological examination. PARTICIPANTS: Volunteers (n = 48 313) initially selected at random from the electoral registers of four cities, subsequently selected at random from questionnaire respondents stratified by answers to questions about hearing. MAIN OUTCOMES MEASURE: Inter-aural hearing threshold level differences measured audiometrically, as a function of age and gender. RESULTS: Tables of inter-aural threshold level differences provided as a resource with potential medicolegal, clinical and research applications. Based on the average of the frequencies 0.5, 1, 2 and 4 kHz, approximately 1% of the general UK population aged 18-80 years have an asymmetry of 15 dB or more. The prevalence is greater in older than in younger people. CONCLUSIONS: Inter-aural threshold differences greater than attributable to measurement error are not uncommon in the adult population, even after screening for conductive hearing loss and substantial noise exposure. They are typically of unknown origin. PMID- 19673979 TI - A retrospective cohort study of nasopharyngeal adenocarcinoma: a rare histological type of nasopharyngeal cancer. AB - OBJECTIVES: The purpose of this study was to investigate the clinical manifestations, Epstein-Barr virus (EBV) serology, and treatment outcome of patients with nasopharyngeal adenocarcinoma (NPAC). DESIGN: A retrospective study of clinical data from consecutive patients with NPAC identified between 1964 and 2000. SETTING: A tertiary cancer center in China. PARTICIPANTS: Forty-eight patients diagnosed with NPAC. MAIN OUTCOME MEASURES: Crosstabs and chi-square test were conducted to study the association of positive VCA-IgA levels among different pathological types of NPAC, and also to compare the proportions of local control rates in patients treated with different modalities. The survival rate was calculated using the Kaplan-Meier method, and the Log Rank test was used to compare the survival rates considering different factors. To balance the distribution bias, a multivariate COX model survival analysis was also performed. RESULTS: Of the 48 NPAC patients identified, 45% presented with cervical metastasis. Pathologically, 58% of NPAC cases were common type and 42% were salivary gland type. The positive rate for the EBV antibody VCA-IgA in all patients was 53% and only 24% in the salivary gland type of NPAC. The overall local control rate and the 5-year disease-free survival rates were 87% and 65% respectively. Patients treated with surgery plus radiotherapy (RT) had a significantly higher 5-year disease-free survival rate than patients receiving RT alone (89%versus 75% respectively) (P = 0.039). Multivariate analysis confirmed that treatment modality was the significant factor influencing patient survival (P = 0.027) and the pathological type was not a factor predicting survival. CONCLUSIONS: Nasopharyngeal adenocarcinoma is a distinct entity in all types of nasopharyngeal carcinoma and EBV serology has limited value in its diagnosis. The combination of surgery and RT should be considered for treatment of early lesions of NPAC. PMID- 19673980 TI - B-cell depletion with rituximab for refractory head and neck Wegener's granulomatosis: a cohort study. AB - OBJECTIVES: This study aimed to evaluate the response of refractory Wegener's granulomatosis affecting the ear, nose and throat and granulomatous eye disease to B-cell depletion with rituximab. DESIGN: A retrospective case note review. SETTING: Tertiary Centre. PARTICIPANTS: All patients who received rituximab for refractory Wegener's granulomatosis affecting the head and neck were included. MAIN OUTCOME MEASURES: Demographic and follow-up data at five time points were recorded. Response was measured using change in the Birmingham Vasculitis Activity Score and prednisolone dose. Secondary outcomes included changes in additional immunomodulators and anti-neutrophil cytoplasm antibodies serology. Adverse events were recorded for the duration of follow-up. RESULTS: Thirty-four patients were included in the analysis. The median age was 47, the male to female ratio was 3 : 2 and the overall median follow-up was 25.5 months. At six months, nine (26%) patients had a partial response, twenty-one (62%) were in remission and four (12%) did not respond. All four non-responders went into remission after a second course of rituximab. Total Birmingham Vasculitis Assessment score decreased after rituximab at all time points (P < 0.001). Four of five patients with retro-orbital involvement responded well to treatment. Two patients were considered secondary failures requiring alternative therapy after an initial response. Adverse events included four major chest infections, two cancers and six infusion reactions. CONCLUSIONS: Our cohort derived considerable benefit from rituximab permitting a reduction in immunosuppressive exposure and prednisolone dose with few major adverse effects. There was an 80% (4/5) response in patients with retro-orbital granulomas. The effect of rituximab was most noticeable in the first 6 months (88% response). PMID- 19673981 TI - Preoperative blood sugar and C-reactive protein associated with persistent discharge after incision and drainage for patients with deep neck abscesses. AB - OBJECTIVES: The aim of this study was to investigate whether preoperative C reactive protein (CRP) and blood sugar correlated with persistent discharge after incision and drainage for patients with deep neck abscesses. STUDY DESIGN: Retrospective data analysis. SETTING: Tertiary referral centre. PARTICIPANTS: A total of 204 patients who underwent operation for deep neck abscess. MAIN OUTCOME MEASURES: Persistent discharge after operation. RESULTS: One hundred seventy patients were included for final analyses. Most of the patients were male (n = 115; 68%) and the average age was 54 years. Using logistic regression analyses, age >55 years [odds ratio (OR): 3.053; P = 0.002], preoperative CRP >15 mg/dL (OR: 2.174; P = 0.027), and preoperative blood sugar >8.3 mmol/L (OR: 3.280; P = 0.001) were independent factors correlated with persistent discharge. CONCLUSIONS: Older age, elevated preoperative CRP level and blood sugar had a statistically significant association with persistent discharge after operation in deep neck abscesses patients. PMID- 19673982 TI - Validation of the Clinical COPD Questionnaire as a psychophysical outcome measure in adult laryngotracheal stenosis. AB - OBJECTIVES: To validate the Clinical Chronic Obstructive Pulmonary Disease Questionnaire (CCQ), a patient-administered instrument developed for bronchopulmonary disease as a disease-specific psychophysical outcome measure for adult laryngotracheal stenosis. DESIGN: Prospective observational study. SETTINGS: Tertiary/National referral airway reconstruction centre. PARTICIPANTS: Thirty-three tracheostomy-free patients undergoing endoscopic laryngotracheoplasty. MAIN OUTCOME MEASURES: CCQ and the Medical Research Council (MRC) Dyspnoea scale, a previously validated but more limited scale, were administered to patients 2 weeks before surgery, preoperatively, and 2 weeks after endoscopic laryngotracheoplasty. Pulmonary function was assessed preoperatively. Internal consistency was assessed with Cronbach alpha statistics and test-retest reliability was determined using intraclass correlation. Correlations between CCQ and MRC scale, and pulmonary function were used to assess convergent and divergent validity respectively. Instrument responsiveness was assessed by correlating total and domain-specific CCQ scores with anatomical disease severity and post-treatment effect size. RESULTS: There were 12 males and 21 females. Mean age was 44 +/- 15 years. Cronbach alpha coefficient and intraclass correlation coefficient were 0.88 and 0.95 respectively. Total and domain-specific CCQ scores significantly correlated with the MRC scores (P < 0.001) and significant correlations between CCQ and peak expiratory flow rate and FEV(1) were identified (P < 0.03). There were statistically significant changes in total and domain-specific CCQ scores when different stenosis severities were compared. Clinical COPD Questionnaire scores also changed significantly and congruently following surgery (P < 0.05 in both cases). DISCUSSION: Clinical COPD Questionnaire is a valid and sensitive instrument for assessing symptom severity and levels of function and well-being in adult patients with laryngotracheal stenosis and can be used as a patient-centred disease-specific outcome measure for this condition. PMID- 19673983 TI - A controlled clinical trial of a therapeutic bacteriophage preparation in chronic otitis due to antibiotic-resistant Pseudomonas aeruginosa; a preliminary report of efficacy. AB - OBJECTIVES: To evaluate the efficacy and safety of a therapeutic bacteriophage preparation (Biophage-PA) targeting antibiotic-resistant Pseudomonas aeruginosa in chronic otitis. DESIGN: Randomised, double-blind, placebo-controlled Phase I/II clinical trial approved by UK Medicines and Healthcare products Regulatory Agency (MHRA) and the Central Office for Research Ethics Committees (COREC) ethical review process. SETTING: A single specialist university hospital. PARTICIPANTS: 24 patients with chronic otitis with a duration of several years (2 58). Each patient had, at the time of entry to the trial, an ear infection because of an antibiotic-resistant P. aeruginosa strain sensitive to one or more of the six phages present in Biophage-PA. Participants were randomised in two groups of 12 treated with either a single dose of Biophage-PA or placebo and followed up at 7, 21 and 42 days after treatment by the same otologist. Ears were thoroughly cleaned on each occasion and clinical and microbiological indicators measured. MAIN OUTCOME MEASURES: Physician assessed erythema/inflammation, ulceration/granulation/polyps, discharge quantity, discharge type and odour using a Visual Analogue Scale (VAS). Patients reported discomfort, itchiness, wetness and smell also using a VAS. Bacterial levels of P. aeruginosa and phage counts from swabs were measured initially and at follow-up. At each visit patients were asked about side effects using a structured form. Digital otoscopic images were obtained on days 0 and 42 for illustrative purposes only. RESULTS: Relative to day 0, pooled patient- and physician-reported clinical indicators improved for the phage treated group relative to the placebo group. Variation from baseline levels was statistically significant for combined data from all clinic days only for the phage treated group. Variation from baseline levels was statistically significant for the majority of the patient assessed clinical indicators only for the phage treated group. P. aeruginosa counts were significantly lower only in the phage treated group. No treatment related adverse event was reported. CONCLUSION: The first controlled clinical trial of a therapeutic bacteriophage preparation showed efficacy and safety in chronic otitis because of chemo resistant P. aeruginosa. PMID- 19673986 TI - The first chapter of an ORL detective novel. PMID- 19673984 TI - A patient with dry mouth. AB - INTRODUCTION: Xerostomia is a common symptom among patients referred to ENT clinics. We present an evidence-based approach in a patient complaining of xerostomia who has not been exposed to radiotherapy. METHOD (search strategy): This review was based on a literature search last performed on 1 July 2008. MEDLINE and EMBASE databases and the Cochrane Library were searched using the subject headings dry mouth, hyposalivation and xerostomia in combination with diagnosis, therapy and surgery. Results were limited to English language articles including clinical trials, randomised controlled trials, meta-analyses, systematic reviews, review articles and human studies. Relevant references from selected articles were reviewed. RESULTS: Xerostomia is not synonymous with hyposalivation. Effective management covers symptomatic relief with selected saliva substitutes, sialogogic agents, addressing underlying dental complications and oral infections, and review of prescribed medication. CONCLUSION: Xerostomia is a common symptom for a wide range of triggering factors, but the treatment is largely palliative and preventative in nature. PMID- 19673987 TI - Paediatric commentary. PMID- 19673988 TI - Consensus statement on management in the UK: transoral laser assisted microsurgical resection of early glottic cancer. AB - BACKGROUND: Transoral laser assisted microsurgical resection of early glottic laryngeal cancer is a relatively new treatment modality that is practised in many centres across the UK. In the absence of the results from randomised clinical trials, clinicians may be guided by an expert panel consensus statement on transoral laser assisted microsurgical resection of early squamous cell cancer of the larynx. OBJECTIVE: To provide consensus recommendations on the various aspects of transoral laser assisted microsurgical resection for early glottic cancer. EVALUATION METHOD: Nine centres across the UK were invited to describe current practice and outcomes for transoral laser assisted microsurgical resection of early glottic cancer. Four working groups were created to draw consensus on standard of care, surgical procedures, outcomes measures and training/certification. The feedback from these groups was integrated into the consensus statement. CONCLUSIONS: The consensus meeting confirmed the stablished and widespread use of transoral laser assisted microsurgical resection for early glottic cancer throughout the UK. The common experience gained allowed a full discussion of all aspects of the management with consensus achieved in key areas of standards of care, surgery, histopathologic reporting, outcomes assessment and training. This consensus statement will result in closer auditing of management and dissemination of results. PMID- 19673989 TI - Rapid umbilical cord diagnostic of hereditary profound hearing loss: how we do it. PMID- 19673990 TI - The assessment of hearing test results following surgery for otitis media with effusion in adults using the Glasgow Benefit Plot: how we do it. PMID- 19673991 TI - Trans-nasal oesphagoscopy: cost implications for a change in practice: how we do it. PMID- 19673992 TI - Management of laryngeal dysplasia in the United Kingdom: a web-based questionnaire survey of current practice. PMID- 19673993 TI - Re: Trends in operative training opportunities for junior and senior trainees in otolaryngology. PMID- 19673995 TI - Re: Aural toilet in infants: how we do it. PMID- 19673997 TI - Re: Hoarse voice in adults: an evidence-based approach to the 12 min consultation. PMID- 19673999 TI - Calculating the risk of occurrence of an adverse outcome given a historical zero numerator. PMID- 19674000 TI - Negative psychosocial consequence of self-restricted driving among cancer survivors in the head and neck region. PMID- 19674001 TI - Tips in tip work for external septorhinoplasty. PMID- 19674002 TI - A survey of ENT out-patient department facilities in district general hospitals across England. PMID- 19674003 TI - Educating patients regarding nasal irrigation using Internet video tutorials. PMID- 19674004 TI - A novel way of cutting Silastic sheeting. PMID- 19674005 TI - What do patients understand by the term 'Vertigo': a cross-sectional study. PMID- 19674006 TI - The BIPP introducer. PMID- 19674007 TI - Closure of nasal septum perforation: a novel technique. PMID- 19674008 TI - Silhouette lift for facial reanimation. PMID- 19674009 TI - Accuracy of clinical coding in ENT day surgery. PMID- 19674010 TI - Frontal sinus approach: the 'vertical bar' concept. PMID- 19674012 TI - The implications of the European Working Time Directive in the medium and smaller sized craft specialties. PMID- 19674013 TI - HLA-B8, DR3: a new risk factor for graft failure after renal transplantation in patients with underlying immunoglobulin A nephropathy. AB - BACKGROUND: The HLA-B8, DR3 haplotype has been associated with high immune reactivity. In this study, we have tested whether this haplotype has differential effect on graft survival in patients with IgAN compared with control patients. METHODS: From the Eurotransplant Registry we analyzed graft survival of 1207 recipients with IgAN and 7935 control patients with non-glomerular diseases. Death-censored graft loss according to the HLA-B8, DR3 haplotype was calculated with Kaplan-Meier analysis and Cox-regression model was used to correct for various risk factors. RESULTS: The frequency of the HLA-B8, DR3 haplotype was significantly lower in IgAN patients compared with controls (10.3% vs. 15.4%, p < 0.001). Ten-year graft survival was identical in the control group with and without the HLA-B8, DR3 haplotype (71.1% and 70.2%, respectively), but significantly worse in IgAN patients carrying the HLA-B8, DR3 haplotype compared with patients without it (52.5% vs. 69.1%, respectively, p = 0.009). The risk of graft loss was increased by 66% (HR 1.6, 95% CI 1.14, 2.29) in IgAN with the HLA B8, DR3 haplotype and independent of well-known risk factors. CONCLUSIONS: We have identified a new risk factor for graft loss unique to patients with IgAN. This finding emphasizes the exclusive immune characteristics of IgAN patients after transplantation. PMID- 19674014 TI - Non-adherence to immunosuppressive medication in renal transplant recipients within the scope of the Integrative Model of Behavioral Prediction: a cross sectional study. AB - BACKGROUND: Non-adherence to immunosuppressive medication is strongly associated with poor outcomes. Identifying the factors influencing it is a first step in developing adherence interventions. This study's objective was to investigate the prevalence of self-reported and collaterally-reported non-adherence to immunosuppressives, and, based on the Integrative Model of Behavioral Prediction, to explore the association between non-adherence, intention to adhere, attitudes, norms and self-efficacy. METHODS: This cross-sectional study included a convenience sample of 114 renal transplant recipients in follow-up care, one to five yr post-transplant. Non-adherence was measured by self-reports and collateral reports. Factors of the Integrative Model of Behavioral Prediction were assessed using a self-report questionnaire. RESULTS: Self-reports showed non adherence of 23.7%; collateral reports showed 3.8%; and a combination of the two showed 26.4%. Logistic regression analysis showed that the attitude "Not all immunosuppressive drugs are necessary to prevent rejection" was less frequent in patients with higher intentions to adhere, with an odds ratio (OR) of 0.05 (95% CI 0.01-0.50). The barrier of "Forgetfulness/Interruption of daily routine" was associated with non-adherence, with an OR of 3.74 (95% CI 1.55-9.03). CONCLUSIONS: Forgetfulness is the most powerful barrier against adherence. Intention to adhere plays a minor role in non-adherence in renal transplant recipients. PMID- 19674015 TI - Quality of life in simultaneous pancreas-kidney transplant recipients. AB - BACKGROUND: Simultaneous pancreas-kidney transplantation (SPK Tx) allows dialysis and insulin therapy to be discontinued and improves the complications of diabetes mellitus type 1 (DM1). This study measure quality of life (QoL) in SPK transplant recipients and determine if there are differences in QoL between these patients and those with DM1 in renal replacement therapy (RRT). METHODS: Short Form Health Survey 36-Item (SF-36) was administered to 69 SPK transplant recipients and 34 patients with DM1 under RRT. A descriptive analysis, multiple linear regression, ANOVA, and ordinal regression (PLUM) models were constructed. RESULTS: QoL was higher in SPK transplant recipients than in patients receiving RRT. The best results were in the recently transplanted patients. Respect to Spanish population the men with SPK transplants scored higher on vitality and lower on general health, role limitations-physical and role limitations-emotional. Women with SPK transplants scored lower on general health. Among patients under RRT, men scored lower on the general health, physical functioning, vitality, and bodily pain while women scored lower on all dimensions. In both groups, greater age was associated with better mental health. CONCLUSION: Positive predictive factors of QoL are SPK Tx and age while negative predictive factors are female sex and RRT. PMID- 19674016 TI - Chronic constipation presenting as acute respiratory distress because of mediastinal shift. PMID- 19674017 TI - Clinical features' diagnostics and treatment of Hirschsprung's disease in adults. AB - AIM: Hirschsprung's disease first diagnosed in adulthood is symptomatic from early childhood in most cases. However, the condition is frequently masked when constipation is managed effectively by enemas and aperients. One third of the patients will experience progressive symptoms or complications, requiring urgent surgical intervention. METHOD: Ninety patients with adult Hirschsprung's disease were observed by a combination of investigations: barium enema, anorectal physiology, estimation of acetylcholinesterase activity and a 60mm full-thickness strip biopsy. The latter two modalities were performed in selected patients where there was diagnostic uncertainty. Single-staged resections, mostly modified Duhamel procedures, were performed in patients who were stable and fit, and with only a limited degree of megacolon present, and no other complications. Otherwise patients were temporarily defunctioned prior to staged resection. All patients underwent surgical treatment. Long-term functional outcomes were expressed as good, satisfactory or poor. RESULTS: Median age was 24.5 years, range 14-47 years, and 72.2% were men. Almost three quarters (73.3%) of the patients had symptoms dating from early childhood. Barium enema was diagnostic in 84.3%, and 36.2% had a positive, but weak rectoanal inhibitory reflex. Acetylcholinesterase staining was positive in 85.7%, but full-thickness strip biopsy was positive in 100% of equivocal cases. A separate cadaveric study of unaffected individuals determined the true length of the normal physiological hypoganglionic zone, mean 24.4 mm and range 7.5-50 mm. Supra-anal short segment Hirschprung's disease was found in 5.6% patients, rectal involvement in 54.4%, rectosigmoid in 38.9% and total aganglionosis of the colon in 1.1%. Resection of the aganglionic zone and proximally dilated colon was performed as a single-staged procedure in 67.8%. Staged surgery was carried out in 32.2%. A modified Duhamel procedure was performed in 91.2% of cases. A good or satisfactory functional outcome was achieved in 96.7%. Long-term functional outcome after resection depends on the degree of preoperative megacolon present. Megacolon limited to the sigmoid colon was associated with a good outcome in 89.7%, but in only 66.7% with more proximal dilatation (P < 0.05). CONCLUSION: Occasionally, Hirschsprung's disease presents in adulthood, with ongoing symptoms from early childhood or with the development of functional obstruction, faecal impaction and megacolon in later life. Diagnosis often requires multimodal investigation. A 60 mm full-thickness strip biopsy confirms aganglionosis in 100%. A modified Duhamel procedure is the operation of choice. PMID- 19674018 TI - Late presentation of Hirschsprung's disease as sigmoid colon volvulus: report of four cases and review of the literature. PMID- 19674019 TI - A systematic review on advancement flaps for rectovaginal fistula in Crohn's disease: transrectal vs transvaginal approach. AB - AIM: Despite advances in the treatment of Crohn's disease (CD), the treatment of rectovaginal (RV) fistula remains challenging. Transrectal (RAF) and transvaginal advancement flaps (VAF) represent two possible alternative surgical approaches to this problem. The study aims to review and compare the results of these approaches for RV fistula in CD. METHOD: Medical databases from January 1983 to August 2008 were consulted for potentially relevant publications. All studies dealing with the RV fistula repair in CD with RAF or VAF were included. Two researchers worked independently on the study selection, quality assessment, data extraction and analysis phases of the study. Analyses were performed with Review Manager 2.0 software. RESULTS: Eleven observational studies were included with a total of 219 flap procedures for RV fistula. The primary fistula closure pooled rate was 54.2% (range 33.3-100%) after RAF and 69.4% (range 0-92.9%) after VAF (P = 0.13). Four studies were eligible for direct comparison between the two procedures. No clearly significant difference between RAF compared with VAF in terms of primary fistula closure rate, nor in terms of overall fistula closure rate, was apparent. The risk of recurrence after RAF compared with VAF seemed similar; in this case, only two studies were taken into consideration. CONCLUSIONS: Although limited by a small number of studies of low clinical evidence level, this systematic review suggests that there is no significant difference in terms of outcome between RAF and VAF for RV fistula in CD. PMID- 19674020 TI - Ethnic disparity in colonic cancer outcomes in New Zealand--biology or an access issue? AB - BACKGROUND: There is an ethnic variation in outcomes for colonic cancer in New Zealand. Whether this disparity is caused by cancer biology or inequitable provision of treatment services after diagnosis has not been elucidated. METHOD: National cancer registry data from 1996 to 2003 were obtained. Incidence and mortality rates for the four major ethnic groups were age-adjusted to the new WHO world population. The impact of age, sex, AJCC stage and site of cancer at diagnosis was compared between ethnic groups using a Cox regression analysis. RESULTS: A total of 11 987 colonic cancer registrations were identified. The overall raw 5-year mortality was 53.7%. The age-adjusted incidence in Europeans was more than double that of the Maori, Asian and Pacific populations at 33.0 per 100,000 population/year. Europeans presented at a greater age, with more right sided cancers, and at an earlier stage of disease. The opposite was true for the Maori population. Pacific Islanders and Asians presented at a younger age, but with a similar site, stage and sex distribution to the rest of the population. There were no significant differences in 5 year mortality after diagnosis when age, sex, stage, and site at presentation were controlled for by cox regression analysis. CONCLUSION: These results suggest that age, sex, stage and site at presentation may be more important than inequality in treatment provision after diagnosis in explaining differences in outcomes between the ethnicities. Efforts need to be focused on identifying reasons for the increased risk of colonic neoplasia in Europeans and the later stage disease presentation in the Maori population. PMID- 19674021 TI - Prospective evaluation of a colorectal cancer nurse follow-up clinic. AB - AIM: Colorectal Nurse Specialist (CNS) clinics for postoperative follow up of colorectal cancer aim to maintain clinical efficacy while reducing costs. We prospectively studied the efficacy and financial implications of such a clinic. METHOD: This was a prospective study of all patients attending CNS clinics over 3 years. A lower-risk protocol for patients with Dukes A was used over 3 years and a higher-risk protocol for patients with Dukes B, C or D was used over 5 years. Department of Health Pricing Charts were used to cost the follow-up protocols, and adjustment was performed to calculate the cost of each quality adjusted life year (QALY) gained. RESULTS: One hundred and ninety-three patients entered into this nurse-led follow-up protocol implemented by the CNS clinic between 2005 and 2007. The Dukes stages and proportions of patients in each stage were as follows: stage A, 13%; stage B, 8%; stage C, 36.3%; and stage D, 9.3%. Ninety-seven per cent underwent curative treatment and 2.6% had palliative treatment. Twenty-one per cent of patients developed recurrent disease. Overall actuarial 5-year survival was 80% and recurrences had a 30% 5-year actuarial survival. The total cost per patient for 3 years of follow up was L1506 and L1179 for lower-risk rectal and nonrectal cancers, respectively. The adjusted cost for each QALY gained for lower-risk tumours was L1914. The total cost per patient with higher risk tumours was L1814 and L1487 for rectal and nonrectal tumours, respectively. The adjusted cost for each QALY gained was L2180 for higher-risk tumours. CONCLUSIONS: This clinic demonstrated cost-effective detection of recurrent disease. Computed tomography (CT) was the most sensitive alert test. As all recurrences were detected within 4 years, we suggest that this is the indicated time to follow up. PMID- 19674022 TI - The prognostic significance of the metastatic lymph node ratio in Dukes stage C colorectal cancer in a district general hospital. AB - AIM: Our aim was to determine if the lymph node ratio would predict overall survival and disease-free survival in Dukes C colorectal cancer in a district general hospital setting in the UK. METHOD: Fifty-six patients were analysed from a prospectively maintained colorectal cancer database. The lymph node ratio was defined as the number of positive lymph nodes divided by the total number of nodes harvested. Comparison was made between the lymph node ratio,TNM nodal status and number of positive lymph nodes by the Kaplan-Meier method. An analysis of covariates was performed by a Cox proportional hazard regression analysis. RESULTS: A lymph node ratio of > 0.25 is prognostically significant for overall survival (P = 0.03) and disease-free survival (P = 0.0003). The lymph node ratio was the strongest covariate in the multivariate regression analysis for recurrence (P = 0.003). CONCLUSION: The lymph node ratio may help clinicians determine which patients have a more aggressive tumour biology and direct appropriate more aggressive chemotherapy regimes towards these patients. PMID- 19674023 TI - HLA-DRB1 alleles are associated with the clinical course of disease and steroid dependence in Mexican patients with ulcerative colitis. AB - AIM: The aim of this study was to study the association between the HLA-DRB1 alleles and the clinical course of ulcerative colitis (UC). METHOD: Seventy five Mexican patients with UC were studied. High resolution HLA typing was performed using Polymerase Chain Reaction-Sequence Specific Oligonucleotide PCR SSO reverse dot blot and Polymerase Chain Reaction-single specific primer PCR SSP. Molecular typing techniques were applied to define HLA-DRB1 alleles. RESULTS: Seventy-five patients (36 female patients, 39 male patients) were studied. Significant associations were found between some HLA-DRB1 alleles and the clinical course of disease: initial active and then inactive and the HLA DRB1*14 allele (P = 0.03; OR = 4.63; 95% CI: 1.08-21.23); and HLA-DRB1*08 allele (P = 0.04; OR = 4.34; 95% CI: 1.9-33.3). On the other hand, the HLA-DRB1*07 (P = 0.001; OR = 9.76 95% CI: 1.55-65.56) was significantly associated with steroid dependence in UC patients. CONCLUSIONS: This study suggests that HLA-DRB1 alleles were associated with the clinical course of disease and steroid dependence in UC patients. PMID- 19674024 TI - Enterocoele is a marker of severe pelvic floor weakness. AB - OBJECTIVE: The aim was to evaluate the relationship between the presence of an enterocoele and grade of rectal prolapse (RP). METHOD: Defaecating proctograms of consecutive patients presenting to the Oxford Pelvic Floor Clinic between January 2004 and November 2008 were analysed. Patients were included if they had full thickness internal (grades 1-4 prolapse) or external RP (grade 5 prolapse). All those included were analysed with regards to the presence of an enterocoele. RESULTS: Three hundred and seventy-one patients [322 (87%) women and 49 (23%) men] were found to have a degree of RP. One out of eight (12.5%) patients with grade 1 RP, 10/42 (18.5%) with grade 2 RP, 34/125 (27%) with grade 3 RP, 62/135 (46%) with grade 4 RP and 23/49 (47%) with grade 5 full thickness external RP had an enterocoele present. This was a statistically significant trend (Pearson chi(2) test P < 0.0002). There was a significantly higher proportion of enterocoeles in women [125/322 (39%) than in men (5/49 (10%)] (P < 0.0001) and a higher likelihood of having an enterocoele with advancing age (P < 0.0001). Within the study, there was no significant difference in the proportion of nulliparous and parous women with enterocoeles (P = 0.8); there were a significantly higher proportion of enterocoeles in hysterectomized women (P = 0.015). CONCLUSIONS: Enterocoele is increasingly seen with advancing RP severity. This suggests that the two findings are part of the same pelvic floor process. These data support the hypothesis that enterocoele is a marker of severe pelvic floor weakness. Enterocoele is seen more frequently in females particularly after hysterectomy. PMID- 19674025 TI - Atrophy of the sphincters of continence in an experimental model. AB - BACKGROUND: Pudendal nerve injury during childbirth may result in external anal sphincter (EAS) atrophy. Recently, balloon compression of the pelvic side wall has been shown to result in EAS atrophy in an experimental model. The aim of this study was to determine whether other sphincters of continence are similarly affected. METHOD: Sixteen adult female virgin Wistar rats (eight controls) were studied 4 weeks after surgery. Anal and urethral canals were dissected, snap frozen and sectioned using a cryostat (100 microm thickness). Masses of EAS, internal anal sphincter (IAS) and external urethral sphincter (EUS) were calculated stereologically and stained with succinate dehydrogenase histochemistry to differentiate striated from smooth muscle. Sphincter length was determined and total sphincter mass calculated. Data were analysed with an unpaired Student's t-test. RESULTS: Atrophy of EAS (30.9 +/- 1.7 to 21.3 +/- 1.7 mg/ kg, P = 0.001), IAS (98.1 +/- 11.3 to 67.1 +/- 5.0 mg/kg, P = 0.01) and EUS (9.6 +/- 0.98 to 7.4 +/- 0.8 mg/kg, P = 0.05) was found 4 weeks after the injury. CONCLUSION: In an experimental model of obstetric pudendal nerve injury, significant atrophy of striated and smooth muscle sphincters of continence occurs and may contribute to altered continence following vaginal childbirth. PMID- 19674026 TI - Ligasure haemorrhoidectomy. PMID- 19674027 TI - MRI for the assessment of locally advanced rectal cancer - a window of opportunity. PMID- 19674028 TI - A prospective multicentre study to investigate percutaneous tibial nerve stimulation for the treatment of faecal incontinence. AB - AIM: Percutaneous tibial nerve stimulation (PTNS) is a minimal invasive treatment that can be performed in the outpatient clinic. This is a pilot study to investigate PTNS in the treatment of faecal incontinence. METHOD: Percutaneous tibial nerve stimulation was performed by insertion of a needle electrode near the posterior tibial nerve. Patients were treated twice a week. Evaluation of faecal incontinence and quality of life was performed at baseline, 6 weeks, 3 months, 6 months and 1 year. Quality of life was estimated using SF-36 and FIQL questionnaires. RESULTS: A total of 22 patients were included. The mean age was 60.4 +/- 11.7 years. After 6 weeks, 18 continued the treatment; 13 patients had a > 50% decrease in incontinence episodes. Overall incontinence episodes fell from 19.6 +/- 21.0 at baseline to 9.9 +/- 15.5 (P = 0.082) at 6 weeks and to 3.6 +/- 4.8 (P = 0.029) at 1 year. Postponement time and quality of life increased significantly during follow up. CONCLUSION: Percutaneous tibial nerve stimulation is simple and can be used in the outpatient setting. Good results can be obtained and sustained during maintenance treatment. PMID- 19674030 TI - Abstracts of the Fourth Annual Meeting of the European Society of Coloproctology. September 23-26, 2009. Prague, Czech Republic. PMID- 19674029 TI - Doppler guided haemorrhoidal arterial ligation with recto-anal-repair (RAR) for the treatment of advanced haemorrhoidal disease. AB - OBJECTIVE: A modification of Doppler guided haemorrhoidal artery ligation (DGHAL) to include the addition of recto-anal repair is reported. Preliminary results of function and safety of third and fourth degree haemorrhoidals are given. METHOD: Thirty patients underwent DGHAL combined with recto-anal-repair (RAR). Each had rectal examination, anorectal manometry and Quality of Life assessment before and 3 months after the procedure. RESULTS: Twenty-nine patients were included in the final analysis. There were three (10.34%) patients of intra-operative and one (3.45%) of postoperative bleeding. Three months after RAR (17.24%) patients with minor residual mucosal prolapse were detected, three (10.34%) patients reported residual symptoms. There was no case of recurrent bleeding. Anal manometry at 3 months after RAR was significantly lower than before the procedure (P < 0.05). One (3.45%) patient reported occasional soiling 3 months after RAR. CONCLUSION: Recto-anal-repair is safe in treating third and fourth degree haemorrhoids with no major complications and low rate of residual disease. PMID- 19674036 TI - Symmetrical primary cutaneous marginal zone lymphoma associated with rheumatoid arthritis. AB - Primary cutaneous marginal zone B-cell lymphoma (PCMZL) is an indolent low grade B cell lymphoma of the skin, with lack of extracutaneous involvement at the time of diagnosis. Herein we report the case of a patient with rheumatoid arthritis (RA) who developed symmetrical PCMZL lesions on both ear lobes. Lesions occurring symmetrically on ear lobes are more specific for cutaneous lymphoid hyperplasia (CLH) and this kind of symmetrical localization hasn't been reported for PCMZL before. PCMZL is considered to arise from a background of reactive lymphoid hyperplasia and this case point out the concept of CLH and PCMZL spectrum. Association of marginal zone lymphoma with rheumatoid arthritis and resolution of lesions together with the resolution of symptoms due to rheumatoid arthritis after rituximab therapy is another interesting point for this case. To the best of our knowledge PCMZL associated with RA has not been reported previously. PMID- 19674037 TI - Contemporary hair transplantation. AB - BACKGROUND: The field of hair transplantation has evolved considerably over the last quarter century. Performed correctly, the cosmetic results of contemporary hair transplantation are virtually undetectable in women and men. Large, pluggy "punch grafts" have been replaced with natural-appearing follicular unit grafts, which maintain their existing anatomy and with proper technique can match the orientation of surrounding hair follicles. OBJECTIVE: To review all of the steps involved in hair transplantation surgery and to provide an overview of medications used in conjunction with transplantation to help prevent hair loss. MATERIALS, METHODS, AND RESULTS: The authors review key aspects of the consultation, physical examination, selection of appropriate candidates, excision of donor area, hairline design, graft creation and placement, and postoperative instructions. The role of medications such as minoxidil and finasteride in preventing ongoing hair loss is an essential part of the treatment plan. For nonsurgical candidates, other treatments such as wigs, hairpieces, and camouflages are reviewed. Future trends may involve the use of low-level laser light therapy, dutasteride, and cloning of follicles. CONCLUSION: Patients and physicians alike are pleased with the results of contemporary hair transplantation, and physicians can now recommend the procedure without reservation. The authors have indicated no significant interest with commercial supporters. PMID- 19674038 TI - Treatment of melasma in Asian skin using a fractional 1,550-nm laser: an open clinical study. AB - BACKGROUND: Melasma is a common hyperpigmentation disorder that can cause refractory cosmetic disfigurement, especially in Asians. Fractional photothermolysis (FP) has been reported to be effective for the treatment of melasma, despite small study populations and short follow-up periods. OBJECTIVE: To evaluate the efficacy and safety of FP for the treatment of melasma in Asians. PATIENTS AND METHODS: Twenty-five patients with melasma received four monthly FP sessions and were followed up to 24 weeks after treatment completion. Efficacy was evaluated using objective and subjective ratings, Melasma Area and Severity Index (MASI), melanin index tracking, and skin elasticity measurements. RESULTS: Investigators observed clinical improvements in 60% and patients in 44% at 4 weeks after treatment, but the figures decreased to 52% and 35%, respectively, at 24 weeks after treatment. Mean MASI scores decreased significantly from 7.6 to 6.2. Mean melanin index decreased significantly after the first two sessions, but it relapsed slightly in subsequent follow-ups. The treatment did not alter skin elasticity. Hyperpigmentation was observed in three of 23 subjects (13%). CONCLUSION: Treatment of melasma with FP led to some clinical improvements, but it was not as efficacious as previously reported at 6-month follow-up. We recommend judicious use of FP for the treatment of melasma in Asian skin because of its limited efficacy. PMID- 19674039 TI - Near-infrared fluorescence venography: a navigation system for varicose surgery. AB - BACKGROUND: Successful varicose vein surgery depends on accurate assessment and identification of veins. We report clinical experiences with a simple, rapid method of detecting varicose veins using indocyanine green (ICG) fluorescence video venography. PATIENTS AND METHOD: ICG percutaneously injected into the great saphenous vein at the ankle or into the ligated end of veins in 20 patients with varicose veins was used as the optical tracer agent. Intraoperative ICG imaging was performed using a laser-fluorescence imaging device. RESULTS: ICG fluorescence venography was performed without any adverse effects. The imaging device provides unparalleled visualization of varicose veins during vein ligation, phlebectomy, and stripping. CONCLUSION: This method enabled easy, safe, and accurate diagnoses, simplified the search for veins during the operation, and minimized surgical invasiveness in varicose vein surgery. The system has great potential as a navigational technique for phlebologic diagnosis and treatment of varicose veins. Further research and development to improve its utility are suggested. PMID- 19674041 TI - Global patterns in belowground communities. AB - Although belowground ecosystems have been studied extensively and soil biota play integral roles in biogeochemical processes, surprisingly we have a limited understanding of global patterns in belowground biomass and community structure. To address this critical gap, we conducted a meta-analysis of published data (> 1300 datapoints) to compare belowground plant, microbial and faunal biomass across seven of the major biomes on Earth. We also assembled data to assess biome level patterns in belowground microbial community composition. Our analysis suggests that variation in microbial biomass is predictable across biomes, with microbial biomass carbon representing 0.6-1.1% of soil organic carbon (r(2) = 0.91) and 1-20% of total plant biomass carbon (r(2) = 0.42). Approximately 50% of total animal biomass can be found belowground and soil faunal biomass represents < 4% of microbial biomass across all biomes. The structure of belowground microbial communities is also predictable: bacterial community composition and fungal : bacterial gene ratios can be predicted reasonably well from soil pH and soil C : N ratios respectively. Together these results identify robust patterns in the structure of belowground microbial and faunal communities at broad scales which may be explained by universal mechanisms that regulate belowground biota across biomes. PMID- 19674042 TI - Photosynthetic picoeukaryote community structure in the South East Pacific Ocean encompassing the most oligotrophic waters on Earth. AB - Photosynthetic picoeukaryotes (PPEs), comprising organisms < 3 mum in size, are important primary producers in marine food webs and include representatives from all known algal lineages. Little is known, however, regarding the composition and distribution of PPE communities, particularly at large spatial scales, or in relation to the underlying biotic and abiotic factors that influence this structure. Here, we analysed PPE community structure along a transect in the South East Pacific Ocean (BIOSOPE cruise) that encompassed a large trophic gradient, including hyper-oligotrophic waters in the South Pacific Gyre (SPG), considered to be some of the 'clearest' natural waters on Earth. Using dot blot hybridizations with 16S rRNA oligonucleotide probes, we established that the PPE community was dominated by members of the classes Prymnesiophyceae and Chrysophyceae throughout the transect. Moreover, clone library construction followed by phylogenetic analysis of sequenced clones revealed several novel 16S rRNA gene lineages, including new clades of prymnesiophytes (designated Prym 16S III) and prasinophytes (Pras 16S-VIII). Pras 16S-VIII was found at all five stations at which clone libraries were constructed, representing a range of trophic conditions, including the South Pacific Gyre, suggesting members of this clade have a broad distribution in this part of the South East Pacific at least. In contrast, Prym 16S-III sequences were largely restricted to oligotrophic stations of the SPG. Subsequent multivariate statistical analyses showed that, within the measured factors, chemical and biological factors seem to influence PPE community structure more than physical parameters. However, more than 50% of the variation in distribution of PPE classes remained unexplained. PMID- 19674043 TI - Language development before and after temporal surgery in children with intractable epilepsy. AB - PURPOSE: To obtain systematic knowledge of language development before and after epilepsy surgery in regions that, if damaged, are known to entail language impairment in adults. METHODS: Twenty-four children (mean age 11 years; range 5.8 15.7 years) with pharmacologically intractable epilepsy participated prior to (the majority) anterior temporal lobectomy and 6, 12, and 24 months thereafter. Reception and production of lexicon (vocabulary) and syntax (sentence structure including grammar) were examined using developmental language tests that provide normative data. RESULTS: Prior to surgery the mean language delay varied from 1.7 years (productive syntax) to 3.5 years (productive lexicon). For lexicon, language delay was larger, the older the children; for syntax it was smaller in children with mothers with higher education. Over the 2 years following surgery, the children developed in all four language components. Development was, however, slower than normal, that is, language delay increased, in three of the four components: in productive lexicon it continued to increase, and in receptive lexicon and productive syntax it appeared to stabilize during the second year. Receptive syntax developed at a normal pace. The development of productive lexicon was remarkably slow when surgery and language mediation were both in the left hemisphere. DISCUSSION: Pharmacologically intractable epilepsy of the temporal lobe, or the underlying condition, is a significant risk factor for delayed language development. Temporal epilepsy surgery does not result in acceleration of language development. If language is still mediated in the operated left hemisphere, development of particular language components may slow down after surgery. PMID- 19674044 TI - Precipitous induction of audiogenic kindling by activation of adenylyl cyclase in the amygdala. AB - PURPOSE: Kindling of audiogenic seizure (AGS) involves >or=14 AGS over 1-2 weeks in genetically epilepsy-prone rats (GEPR-9s) and induces gradual seizure duration increases, epileptiform electroencephalography (EEG), and emergence of post tonic clonus (PTC), which are long-lasting. N-methyl-d-aspartate (NMDA)-receptor activation in lateral amygdala (LA) is implicated in AGS kindling initiation. However, the persistence of AGS kindling appears to be dependent on molecular mechanisms initiated by NMDA-receptor activation, which may involve adenylyl cyclase (AC). This study attempted to mimic AGS kindling persistently in nonkindled GEPR-9s by one-time activation of AC in LA. METHODS: The effects of a single focal bilateral microinjection into LA of an AC activator, MPB forskolin {7-Deacetyl-7-[O-(N-methylpiperazino)-gamma-butyryl]-forskolin dihydrochloride} (25-100 pmol/side), on seizure behavior in GEPR-9s were evaluated. RESULTS: One time bilateral microinjection of MPB forskolin in GEPR-9s precipitously induced an AGS kindling-like effect, which involved significant increases in seizure duration and long-lasting susceptibility to AGS that culminates in PTC. This effect occurred at 24 h after MPB forskolin microinjection and lasted >or=5 weeks. The effect was seen when AGS was initiated at 1 and 12 h after microinjection, but not if AGS was induced only at 24 h, indicating the importance of the temporal proximity of AGS induction to the MPB forskolin microinjection. DISCUSSION: These findings indicate that one-time activation of AC within the NMDA receptor-mediated molecular cascade results in precipitous induction of AGS kindling. These data further suggest that AC activation in the LA may be an important epileptogenic mechanism that subserves the long-lasting persistence of AGS kindling. PMID- 19674045 TI - Improved seizure control by alternating therapy of levetiracetam and valproate in epileptic rats. AB - PURPOSE: Tolerance to drug treatment is a serious problem in the treatment of epilepsy. We previously showed that tolerance to levetiracetam (LEV) developed within 4 days after the start of the treatment in a rat model for spontaneous seizures after electrically induced status epilepticus. In the current study we tested whether the development of tolerance to LEV could be prevented by alternating between LEV and valproate (VPA) treatment. METHODS: Before starting the alternating therapy with LEV and VPA (3 day LEV-3 day VPA, two cycles), we assessed the efficacy of VPA monotherapy by administering VPA to chronic epileptic rats via osmotic minipumps during 7 days. The anticonvulsive effects were determined by continuous video-EEG (electroencephalography) monitoring, and the concentration of VPA and LEV was measured in plasma using gas chromatography. RESULTS: VPA significantly suppressed spontaneous seizures in chronic epileptic rats for 5 days. Hereafter, seizure frequency increased to pretreatment values despite adequate VPA blood levels. Seizure duration was reduced for 6 days during treatment. Seizure severity was reduced throughout the 7-day treatment period. Alternating treatment of LEV and VPA did not prevent development of tolerance; however, seizures were suppressed significantly longer compared to VPA and LEV monotherapy. CONCLUSIONS: Because alternating treatment with LEV and VPA led to a prolonged effective seizure control in the animal model, it would be worthwhile to explore the possibilities of using an alternating treatment protocol in pharmacoresistant patients in whom an effective treatment is hampered by tolerance to antiepileptic drugs. PMID- 19674046 TI - Spike-wave discharges are necessary for the expression of behavioral depression like symptoms. AB - PURPOSE: The WAG/Rij strain of rats, a well-established model for absence epilepsy, has comorbidity for depression. These rats exhibit depression-like behavioral symptoms such as increased immobility in the forced swimming test and decreased sucrose intake and preference (anhedonia). These depression-like behavioral symptoms are evident in WAG/Rij rats, both at 3-4 and 5-6 months of age, with a tendency to aggravate in parallel with an increase in seizure duration. Here we investigated whether the behavioral symptoms of depression could be prevented by the suppression of absence seizures. METHODS: Ethosuximide (ETX; 300 mg/kg/day, in the drinking water) was chronically applied to WAG/Rij rats from postnatal day 21 until 5 months. Behavioral tests were done before the cessation of the treatment. Electroencephalography (EEG) recordings were made before and after cessation of treatment to measure seizure severity at serial time-points. RESULTS: ETX-treated WAG/Rij rats exhibited no symptoms of depression-like behavior in contrast to untreated WAG/Rij rats of the same age. Moreover, treated WAG/Rij rats did not differ from control age-matched Wistar rats. ETX treatment led to almost complete suppression of spike-wave discharges (SWDs) in 5-6 month old WAG/Rij rats. Discontinuation of chronic treatment was accompanied by a gradual emergence of SWDs; however, a persistent reduction in seizure activity was still present 47 days after discontinuation of the chronic treatment. DISCUSSION: The results suggest that seizure activity is necessary for the expression of depression-like behavioral symptoms and confirm that epileptogenesis can be prevented by early and chronic treatment. PMID- 19674047 TI - Histologic and morphologic effects of valproic acid and oxcarbazepine on rat uterine and ovarian cells. AB - PURPOSE: To determine the histologic and morphologic effects of valproic acid (VPA) and oxcarbazepine (OXC) on rat uterine and ovarian cells. METHODS: Fifty six female prepubertal Wistar rats (21-24 days old and weighing between 47.5 and 58.1 g) were divided equally into four groups, which were given drinking water (controls), 300 mg/kg/day of VPA, 100 mg/kg/day of OXC or VPA + OXC via gavage, for 90 days. Ovaries and uteri of rats on proestrous and diestrous phases of estrous cycle were extirpated and placed in a fixation solution. The tissue specimens were assessed with apoptosis (TUNEL) staining protocols, eosinophil counting, and electron microscopic techniques. RESULTS: In uteri, apoptosis in stroma, mitochondrial swelling, and cristolysis were observed in the VPA group, and OXC led to negative effects on epithelial cell and intracellular edema. In ovaries, both drugs increased apoptosis and intracytoplasmic edema. Organelle structure disruption was also observed in the OXC group. More conspicuous degenerative modifications were determined in the VPA + OXC group. In uteri, the number of TUNEL-positive luminal epithelial cells was 7.20 +/- 1.32 in controls, and significantly increased to 29.60 +/- 1.58, 34.20 +/- 2.53, and 54.80 +/- 2.04 in VPA, OXC, and VPA + OXC groups, respectively (p < 0.001). The highest number of TUNEL-positive glandular epithelium cells was observed in the VPA + OXC group; however, the number of TUNEL-positive stroma cells was highest in the VPA group. The highest number of eosinophils in stroma was in the VPA group. CONCLUSION: VPA and OXC trigger apoptotic and degenerative effects on rat uterine and ovarian cells. VPA also prevents implantation of embryo to the uterus and causes abortion via endometrial eosinophil infiltration. PMID- 19674048 TI - Spatial distribution of intracranially recorded spikes in medial and lateral temporal epilepsies. AB - PURPOSE: Although seizures and interictal spikes are not always colocalized, there may be valuable localizing information in the spatial distribution of spikes. To test this hypothesis, we studied the spatial distribution of intracranially recorded interictal spikes in patients with medial temporal (MT) and lateral temporal (LT) neocortical seizure onset. METHODS: A total of 21 patients (MT n = 12, LT n = 9) who completed intracranial monitoring were selected for this study. Two 4-h intracranial electroencephalography (icEEG) epochs were analyzed, one during wake and one during sleep, both at least 6 h removed from seizures. Spikes detected automatically in medial temporal structures [hippocampal formation (H) and entorhinal cortex (EC)], and in five cortical areas (occipital, frontal, parietal, lateral temporal, and inferior temporal) were tabulated. RESULTS: Interictal spikes occurred broadly over medial temporal structures and cortical areas in MT and LT patients. The patients differed with a greater number of spikes in medial temporal structures in the MT group (p < 0.05 for H and p < 0.001 for EC) and a greater number of spikes in parietal (p < 0.01) and frontal (p < 0.001) areas in the LT group. There were sleep-related increases in spike rates in inferomedial temporal structures in both groups. The two groups could be separated with a classifier based on medial temporal and parietal and frontal spikes (p < 0.0001). DISCUSSION: MT and LT patients have different spatial distributions of interictal spikes and can be distinguished by the relative spike rates in medial temporal and extratemporal areas during sleep and wake. PMID- 19674049 TI - The impact of diazepam's discovery on the treatment and understanding of status epilepticus. AB - The fortuitous discovery of the benzodiazepines and the subsequent application of these agents to the treatment of status epilepticus (SE) heralds in the modern age of treating this neurologic emergency. More than 50 years after their discovery, the benzodiazepines remain the drugs of first choice in the treatment of SE. However, the benzodiazepines can be ineffective, especially in those patients whose seizures are the most prolonged. The benzodiazepines act by increasing the affinity of gamma-aminobutyric acid (GABA) for GABAA receptors. A receptor's subunit composition affects its functional and pharmacologic properties, trafficking, and cellular localization. The GABAA receptors that mediate synaptic inhibition typically contain a gamma2 subunit and are diazepam sensitive. Among the GABAA receptors that mediate tonic inhibition are the benzodiazepine-insensitive delta subunit-containing receptors. The initial studies investigating the pathogenesis of SE demonstrated that a reduction in GABA-mediated inhibition within the hippocampus was important in maintenance of SE, and this reduction correlated with a rapid modification in the postsynaptic GABAA receptor population expressed on the surface of the hippocampal principal neurons. Subsequent studies found that this rapid modification is, in part, mediated by an activity-dependent, subunit-specific trafficking of the receptors that resulted in the reduction in the surface expression of the benzodiazepine sensitive gamma2 subunit-containing receptors and the preserved surface expression of the benzodiazepine-insensitive delta subunit-containing receptors. This improved understanding of the changes in the trafficking of GABAA receptors during SE partially accounts for the development of benzodiazepine pharmacoresistance and has implications for the current and future treatment of benzodiazepine-refractory SE. PMID- 19674050 TI - Successful treatment for refractory convulsive status epilepticus by non parenteral lacosamide. AB - Lacosamide (Vimpat) is a newly licensed novel antiepileptic drug. We report a case of refractory convulsive status epilepticus (CSE) that was successfully controlled with lacosamide. The 38-year-old male patient was admitted for a series of complex partial seizures with secondary generalization leading to refractory CSE. During the transport to the hospital the patient was given 22.5 mg diazepam, 12.5 mg etomidate, and 5 mg midazolam without success. An additional dose of 4 mg lorazepam and a dose of 1,500 mg levetiracetam after admission were yet without clinical effect. A further treatment with lacosamide (300 mg via percutaneous gastric fistula) resulted in complete clinical remission of the epileptic activity within 30 min. The application of lacosamide resulted in cessation of CSE and was well tolerated. To our knowledge, this is the first case of successful treatment of refractory CSE with lacosamide. Further studies are needed to evaluate the safety and efficacy of lacosamide in treatment of SE. PMID- 19674051 TI - Parkinsonism induced by VNS in a child with double-cortex syndrome. AB - We describe a child with epilepsy associated with double-cortex syndrome in whom vagus nerve stimulation (VNS) generated parkinsonian symptoms. A 13-year-old girl presented with refractory secondary generalized epilepsy from the age of 6 years and mental retardation. Her electroencephalography (EEG) showed diffuse polyspike and wave discharges. Magnetic resonance imaging (MRI) showed double-cortex syndrome. She was submitted to extended callosal section at the age of 10 years, which yielded 50% seizure frequency reduction. She was submitted to VNS by the age of 12 years. As stimulation intensity was increased, there was appearance of extrapyramidal symptoms: She developed bilateral tremor and rigidity, and gait and postural disturbance. All symptoms disappeared 7-10 days after VNS was turned off. Several attempts to reactivate VNS led to the same results. During the periods when VNS was on she presented with marked seizure frequency reduction. This is the first report of a clinically evident direct effect of VNS on the basal ganglia. PMID- 19674052 TI - Balance impairment in chronic antiepileptic drug users: a twin and sibling study. AB - PURPOSE: Patients taking antiepileptic drugs (AEDs) have an increased incidence of fractures. This study investigated chronic AED use and physical contributors to falls risk using an AED-discordant, twin and sibling matched-pair approach, and assessed clinically relevant subgroups: AED polytherapy; longer-duration AED; and falls history. METHODS: Twenty-nine same-sex (mean age 44.9 years, 59% female), ambulatory, community-dwelling twin and sibling pairs, discordant for AED exposure (and AED-indication), were recruited. Validated clinical and laboratory tests of strength, gait, and balance were performed. Relevant AED levels, and fasting serum samples for 25-hydroxyvitamin D (25OHD), 1,25 dihydroxyvitamin D [1,25(OH)(2)D], and immunoreactive parathyroid hormone (iPTH) levels were taken. RESULTS: There were significant mean within-pair differences in tests of static and dynamic balance, with the AED user having poorer balance function than the AED nonuser. No difference was seen in lower limb strength or gait measures. Increased duration of AED therapy and AED polytherapy were independent predictors of increased sway. No significant within-pair differences were seen in fasting serum levels of 1,25(OH)(2)D, 25OHD and iPTH after Bonferroni correction. DISCUSSION: Balance performance is impaired in AED users compared to their matched nonuser siblings. Pairs where the AED users took AED polytherapy, or had a longer duration of AED use, had more impaired balance performance. These balance deficits may contribute to the increased rate of fractures in this population. PMID- 19674053 TI - Optical suppression of experimental seizures in rat brain slices. AB - PURPOSE: To determine if a small ultraviolet emitting diode (UV LED) could release sufficient gamma-aminobutyric acid (GABA) from a caged precursor to suppress paroxysmal activity in rat brain slices. METHODS: Electrophysiologic recordings were obtained from rat brain slices bathed with caged GABA: 4-[[(2H benzopyran-2-one-7-amino-4-methoxy)carbonyl]amino]butanoic acid (BC204), at concentrations between 3 and 30 microm. Seizure-like activity was induced by perfusing slices with extracellular medium lacking magnesium and containing 4 aminopyridine (4-AP; 100 microm). A small, high-power UV LED was used to uncage BC204 and determine whether an increase in ambient GABA could alter normal or paroxysmal activity in the slice. RESULTS: UV LED illumination, in the absence of BC204, had no effect on CA1 population spikes or seizure-like activity. The light did induce a small temperature elevation (<0.15 degrees C) over the current intensities and exposure durations used in these experiments. In the presence of BC204, UV light decreased the CA1 population spike and seizure-like activity. The BC204 effect can be best accounted for by release of GABA: The reduction of population spikes and seizure-like activity was blocked by the GABA antagonist picrotoxin, and BC204 illumination produced a membrane polarization that reversed at the expected potential for GABA(A) receptors. DISCUSSION: These experiments establish that illumination of a low concentration of caged GABA with a tiny UV LED can release sufficient GABA to attenuate seizure-like activity in brain slices. Because our seizure model is very severe, it is probable that this technique would have a robust effect in human focal epilepsy. PMID- 19674054 TI - Incidence of newly diagnosed epileptic seizures in a French South Indian Ocean Island, La Reunion (EPIREUN). AB - PURPOSE: To assess the incidence of newly diagnosed epileptic seizures in the population of La Reunion. METHODS: From July 1, 2004 to June 30, 2005, we conducted a prospective, observational, and multicenter epidemiologic study to identify patients with newly diagnosed epileptic seizures. Febrile and neonatal seizures were excluded. RESULTS: Seven hundred sixty-six patients were included. The standardized (2000 U.S. population) incidence rate of all suspected cases of newly diagnosed (provoked and unprovoked) epileptic seizures was 115.4/100.000 person-years [95% confidence interval (CI) 106.7-124.0]. We observed a bimodal distribution: The crude incidence was 99.5/100,000 in the group aged 0-14 years and 330.8/100,000 in those older than 65 years. One hundred thirty-five cases were classified as provoked seizures (17.6%; incidence 17.7/100,000). Alcohol consumption, cranial trauma, and cerebrovascular disease were the most frequent causes (27.4%, 11.1%, and 10.4%, respectively). Six hundred twenty cases were classified as unprovoked seizures (single and recurrent) (80.9%; incidence, 81.2/100,000). Two hundred sixty cases of seizures were due to stable neurologic conditions (incidence, 34.1/100,000) and the most common causes were cerebrovascular disease (46.2%), alcoholism (20.4%), and cranial trauma (5.4%). Evolutive neurologic conditions contributed to 23 cases (incidence, 3.0/100,000). Lastly, unprovoked seizures with unknown etiology were 337 (incidence, 44.2/100,000). CONCLUSIONS: The global incidence rate of newly diagnosed epileptic seizures in La Reunion was clearly higher than those observed in industrialized countries and similar to those observed in developing countries. The major risk factors were represented by cerebrovascular disease, alcohol consumption, and cranial trauma. Surprisingly, there were few infections. PMID- 19674055 TI - Protective effects of naloxone in two-hit seizure model. AB - PURPOSE: Early life status epilepticus (SE) could enhance the vulnerability of the immature brain to a second SE in adulthood (two-hit seizure model). Naloxone has been proved to possess inflammation inhibitory effects in nervous system. This study was designed to evaluate the dose-dependent protective effects of naloxone in kainic acid (KA)-induced two-hit seizure model. METHODS: After KA induced SE at postnatal day 15 (P15), Sprague-Dawley rats were infused with either saline or different doses (1.92, 3.84, 5.76, and 7.68 mg/kg) of naloxone continuously for 12 h. De novo synthesis of cytokines (interleukin-1 beta [IL-1 beta], S100B) was assessed by real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) at 12 h after P15 SE. Glial activation states were analyzed by western blotting of glial markers (glial fibrillary acidic protein [GFAP], S100B, Iba1) both at 12 h after P15 SE and at P45. After a second SE at P45, cognitive deteriorations were evaluated by Morris water tests and neuron injuries were evaluated by TdT-mediated dUTP nick end labeling (TUNEL) assays. RESULTS: Naloxone reduced IL-1 beta synthesis and microglial activation most potently at a dose of 3.84 mg/kg. Attenuation of S100B synthesis and astrocyte activation were achieved most dramatically by naloxone at a dose of 5.76 mg/kg, which is equal to the most powerful dose in ameliorating cognitive injuries and neuron apoptosis after second SE. CONCLUSIONS: Naloxone treatment immediately after early life SE could dose-dependently reduce cytokine production, glial activation, and further lower the vulnerability of immature brains to a second hit in adulthood. PMID- 19674056 TI - Reevaluating the mechanisms of focal ictogenesis: The role of low-voltage fast activity. AB - The mechanisms that control the transition into a focal seizure are still uncertain. The introduction of presurgical intracranial recordings to localize the epileptogenic zone in patients with drug-resistant focal epilepsies opened a new window to the interpretation of seizure generation (ictogenesis). One of the most frequent focal patterns observed with intracranial electrodes at seizure onset is characterized by low-voltage fast activity in the beta-gamma range that may or may not be preceded by changes of ongoing interictal activities. In the present commentary, the mechanisms of generation of focal seizures are reconsidered, focusing on low-voltage fast activity patterns. Experimental findings on models of temporal lobe seizures support the view that the low voltage fast activity observed at seizure onset is associated with reinforcement and synchronization of inhibitory networks. A minor role for the initiation of the ictal pattern is played by principal neurons that are progressively recruited with a delay, when inhibition declines and synchronous high-voltage discharges ensue. The transition from inhibition into excitatory recruitment is probably mediated by local increase in potassium concentration associated with synchronized interneuronal firing. These findings challenge the classical theory that proposes an increment of excitation and/or a reduction of inhibition as a cause for the transition to seizure in focal epilepsies. A new definition of ictogenesis mechanisms, as herewith hypothesized, might possibly help to develop new therapeutic strategies for focal epilepsies. PMID- 19674058 TI - Bitemporal form of partial reading epilepsy: further evidence for an idiopathic localization-related syndrome. AB - Idiopathic partial reading epilepsy (RE) is a rare syndrome. We report the clinical and electroencephalographic characteristics of two right-handed patients with the following: reading-induced independent bilateral temporal lobe seizures, accompanied by alexia in left (dominant) sided seizures recorded on video-EEG (electroencephalography); subclinical activation over left posterior temporal and occipital electrodes during reading; no spontaneous seizure and no other trigger than reading; onset in adolescence; and history of varying resistance to treatment. Bilateral independent temporal lobe reflex seizures are part of the clinical spectrum of RE. It may result from hyperexcitability of bilateral cortical networks involved in the early steps of the reading process. PMID- 19674057 TI - A novel mouse model for sudden unexpected death in epilepsy (SUDEP): role of impaired adenosine clearance. AB - Sudden unexpected death in epilepsy (SUDEP) is a significant cause of mortality in people with epilepsy. Two postulated causes for SUDEP, cardiac and respiratory depression, can both be explained by overstimulation of adenosine receptors. We hypothesized that SUDEP is a consequence of a surge in adenosine as a result of prolonged seizures combined with deficient adenosine clearance; consequently, blockade of adenosine receptors should prevent SUDEP. Here we induced impaired adenosine clearance in adult mice by pharmacologic inhibition of the adenosine removing enzymes, adenosine kinase and deaminase. Combination of impaired adenosine clearance with kainic acid-induced seizures triggered sudden death in all animals. Most importantly, the adenosine receptor antagonist caffeine, when given after seizure onset, increased survival from 23.75 +/- 1.35 min to 54.86 +/ 6.59 min (p < 0.01). Our data indicate that SUDEP is due to overactivation of adenosine receptors and that caffeine treatment after seizure onset might be beneficial. PMID- 19674059 TI - Seizures increase cell proliferation in the dentate gyrus by shortening progenitor cell-cycle length. AB - PURPOSE: A prolonged seizure, status epileptics (SE), is a potent stimulus for increased neurogenesis in the dentate gyrus of the hippocampus. Molecular mechanisms that regulate normal and pathologic cell birth in the dentate gyrus are poorly understood. METHODS: Lithium-pilocarpine was used to induce SE in immature postnatal day 20 rats. Newborn cells in the dentate were labeled with bromo-deoxyuridine to determine a time-course of cell proliferation, and measure cell-cycle length. In addition, we studied expression by Western blot and immunohistochemistry of two known inhibitors of G(1)-S cell-cycle progression P27/Kip1 and P15/Ink4b following SE. RESULTS: Cell proliferation in the dentate gyrus increases starting 2 h after SE and is sustained for 40 days. Increased cell proliferation following SE is associated with a shortened dentate gyrus progenitor's cell cycle, 15 h in control to 12 h in the SE animals. To identify molecules responsible for the shortened progenitor cell cycle we studied inhibitors of cell-cycle progression P27/Kip1, and P15/Ink4b. We find decreased phosphorylation at P27/Kip1 Serine 10 and Threonine 187 following SE. Although total P27/Kip1 and P15/Ink4b levels were not altered after SE, P27/Kip1 immunoreactivity was minimal in newborn but increased with maturation of the dentate granule neurons. DISCUSSION: The sustained increase in dentate gyrus cell proliferation following SE provides a large pool of immature dentate granule cells prior to development of spontaneous seizures. A decrease in cell-cycle length of dentate gyrus progenitors is at least partially responsible for increased numbers of newborn cells following SE. PMID- 19674060 TI - Lateralizing language with magnetic source imaging: validation based on the Wada test. AB - PURPOSE: Magnetoencephalography (MEG)/magnetic source imaging (MSI) is a noninvasive functional neuroimaging procedure used to localize language-specific regions in the brain. The Wada test, or intracarotid amobarbital procedure (IAP), is the gold standard in determining speech/language lateralization for presurgical planning, although it is invasive and associated with morbidity. The purpose of this study is to provide further validation on the use of MSI for presurgical language lateralization by comparing results against the IAP. METHODS: The sample consisted of 35 patients with epilepsy and/or brain tumor undergoing presurgical evaluation at the Minnesota Epilepsy Group. All patients received both an IAP and MSI to determine hemispheric language dominance. For MSI, a 148-channel MEG system was used to record activation of language-specific cortex by an auditory word-recognition task. RESULTS: The MSI and IAP were concordant in determining language in the hemisphere to be treated in 86% of the cases with sensitivity and specificity values of 80% and 100%, respectively. CONCLUSIONS: The results from this study are consistent with prior research findings comparing functional neuroimaging procedures to the IAP in determining language lateralization in presurgical patients. The current study provides an important replication and support for Papanicolaou et al.'s findings in 2004 using a consecutive clinical sample from a different institution. An unusually high rate of atypical IAP language cases in this sample and differences between the two procedures are believed to explain the noted discrepancies. MSI is a viable noninvasive alternative to the IAP in the presurgical determination of language lateralization. PMID- 19674061 TI - Evaluation of stereoselective anticonvulsant, teratogenic, and pharmacokinetic profile of valnoctylurea (capuride): a chiral stereoisomer of valproic acid urea derivative. AB - PURPOSE: The purpose of this study was to evaluate the stereoselective anticonvulsant activity, neurotoxicity, pharmacokinetics, and teratogenic potential of two stereoisomers of valnoctylurea (VCU), a central nervous system (CNS)-active urea derivative of valnoctic acid, which is a constitutional isomer of valproic acid (VPA). METHODS: VCU stereoisomers (2S,3S)-VCU and (2R,3S)-VCU were synthesized. Their anticonvulsant activity was determined and compared to VPA and racemic-VCU in rats utilizing the maximal electroshock seizure (MES) and the subcutaneous pentylenetetrazole (scMet) tests. Neurotoxicity was determined in rats using the positional sense test, muscle tone test, and gait and stance test. The induction of neural tube defects (NTDs) by VCU stereoisomers was evaluated in a mouse strain highly susceptible to VPA-induced teratogenicity. The pharmacokinetics of VCU was studied in a stereoselective manner following intraperitoneal (i.p.) administration to rats. RESULTS: (2S,3S)-VCU and (2R,3S) VCU median effective dose ED(50) values were 29 mg/kg [95% confidence interval (CI) = 8-60 mg/kg] and 42 mg/kg (95% CI = 36-51 mg/kg) (MES) and 22 mg/kg (95% CI = 13-51 mg/kg) and 12 mg/kg (95%CI = 7-21 mg/kg) (scMet), respectively. (2S,3S) VCU was more potent and had a wider safety margin (p < 0.05), defined as the protective index (PI = TD(50)/ED(50)), at both the MES (PI > 17) and scMet (PI > 23) tests than racemic-VCU or (R,S)-VCU (PI = 2.8 and 9.9, respectively). VCU stereoisomers caused NTDs at doses >4 times that of their anticonvulsant ED(50) values. At a dose of 112 mg/kg, (2R,3S)-VCU was nonteratogenic and less embryotoxic than its stereoisomer (2S,3S)-VCU. No stereoselective pharmacokinetics was observed following intraperitoneal dosing of racemic-VCU to rats. VCU was mainly eliminated by metabolism with a half-life of 2 h. CONCLUSIONS: VCU anticonvulsant activity and wide PI values make it a potential candidate for development as a new, potent antiepileptic drug (AED). PMID- 19674062 TI - Evidence of shared genetic risk factors for migraine and rolandic epilepsy. AB - PURPOSE: Evidence for a specific association between migraine and rolandic epilepsy (RE) has been conflicting. Children with migraine frequently have electroencephalographic (EEG) abnormalities, including rolandic discharges, and approximately 50% of siblings of patients with RE exhibit rolandic discharges. We assessed migraine risk in RE probands and their siblings. METHODS: We used cohort and reconstructed cohort designs to respectively assess the relative risk of migraine in 72 children with RE and their 88 siblings using International Classification of Headache Disorders (ICHD-2) criteria. Incidences were compared in 150 age and geographically matched nonepilepsy probands and their 188 siblings. We used a Cox proportional hazards model, using age as the time base, adjusting hazard ratios (HRs) for sex in the proband analysis, and for sex and proband migraine status in the sibling analysis. RESULTS: Prevalence of migraine in RE probands was 15% versus 7% in nonepilepsy probands, and in siblings of RE probands prevalence was 14% versus 4% in nonepilepsy siblings. The sex-adjusted HR of migraine for an RE proband was 2.46 [95% confidence interval (CI) 1.06 5.70]. The adjusted HR of having > or =1 sibling with migraine in an RE family was 3.35 (95% CI 1.20-9.33), whereas the HR of any one sibling of a RE proband was 2.86 (95% CI 1.10-7.43). DISCUSSION: Migraine is strongly comorbid in RE and independently clusters in their siblings. These results suggest shared susceptibility to migraine and RE that is not directly mediated by epileptic seizures. Susceptibility gene variants for RE may be tested as risk factors for migraine. PMID- 19674063 TI - Successful bortezomib-based treatment in POEMS syndrome. PMID- 19674067 TI - The CST3 B haplotype is associated with frontotemporal lobar degeneration. AB - BACKGROUND AND PURPOSE: Frontotemporal lobar degeneration (FTLD) is a common cause of early-onset dementia. Given the role of cystatin C in brain neurodegeneration and neuroregeneration, the aim of this study was to determine whether the cystatin C gene (CST3) was genetically associated with FTLD. METHODS: Hundred and eighty-six FTLD patients and 457 controls underwent CST3 analysis by PCR and KspI enzyme digestion. RESULTS: In FTLD patients negative for the presence of PGRN mutations, we found an over-representation of the CST3 haplotype B [odds ratio (OR = 1.619, P = 0.002)] and of AB/BB genotypes (OR = 1.704, P = 0.008) in FTLD patients. CONCLUSIONS: The present study indicated the CST3 B haplotype as a putative risk factor for FTLD in PGRN mutations negative patients. The reduced level of cystatin C, previously associated with the B haplotype, might represent the molecular factor responsible for the increased risk. Long term depletion of neurotrophic factors, such as cystatin C and progranulin proteins, seem to be a common theme in FTLD: boosting the expression of such proteins might be a promising therapeutic strategy for FTLD. PMID- 19674066 TI - Calbindin-1 association and Parkinson's disease. AB - BACKGROUND AND PURPOSE: Calcium levels have been proposed to play an important role in the selective vulnerability of nigrostriatal dopaminergic neurons in Parkinson's disease (PD). Recently, an association was reported between the calcium buffer, calbindin (rs1805874) and risk of PD in a Japanese patient control series. METHODS: We genotyped rs1805874 in four independent Caucasian patient-control series (1543 PD patients, 1771 controls). RESULTS: There was no evidence of an association between rs1805874 and disease risk in individual populations or in the combined series (odds ratio: 1.04, 95% CI: 0.82-1.31, P = 0.74). DISCUSSION: Our study shows there is no association between rs1805874 and risk for PD in four Caucasian populations. This suggests the effect of calbindin on PD risk displays population specificity. PMID- 19674068 TI - Open muscle biopsy in suspected myopathy: diagnostic yield and clinical utility. AB - BACKGROUND: The purpose of the study was to investigate the diagnostic yield and clinical utility of open muscle biopsy and to identify pre-biopsy factors that might predict useful clinical results for suspected myopathy. METHODS: Two hundred fifty-eight muscle biopsies, performed for investigation of suspected myopathy, were evaluated. RESULTS: A specific clinical diagnosis following muscle biopsy was made in 43% of cases. As a result of the biopsy, clinical diagnosis was changed in 47% and treatment was changed in 33% of cases. Results either led to a specific clinical diagnosis or changed the diagnosis/treatment in 74% of patients. Positive family history of myopathy and findings of myopathic irritability on electromyography had a negative predictive value for diagnosis change. CONCLUSIONS: Open muscle biopsy is useful in myopathy evaluation in the modern genetic era. PMID- 19674069 TI - F-waves in lumbosacral radiculopathies: its time has come. PMID- 19674070 TI - A review of 61 cases of ocular tumours seen over a 3-year period in Benin City, Nigeria: a 3-year descriptive retrospective study (1998-2000). AB - The aim of this study is to ascertain the tumour type, sex variation and mean age of presentation of ocular tumours, as well as to update the available literature on the prevalence of the common ocular neoplasms in our environment. In this study, we analysed the data of 61 ocular biopsy specimens received between January 1998 and December 2000 at the Morbid Anatomy Department of the University of Benin Teaching Hospital, Benin City, Nigeria. Slides from paraffin embedded blocks of all ocular biopsies received were reviewed to identify the tumour types, sex variation and age of presentation. PMID- 19674071 TI - Familial gastric cancer and Li-Fraumeni syndrome. AB - Gastric cancer occurs in some familial diseases with inherited cancer predisposition. Genetic factors have been correlated with the hereditary diffuse gastric cancer and other familial gastric cancer conditions as hereditary non polyposis colorectal cancer and Li-Fraumeni syndrome. The present study was aimed at searching for germ line mutations of TP53 gene in familial gastric cancer with cluster for Li-Fraumeni syndrome or Li-Fraumeni-like syndrome. Twenty-three pedigrees with characteristics for Li-Fraumeni-like syndrome were identified. DNA of the proband was sequenced using polymerase chain reaction/single-strand conformation polymorphism. Among these 23 cases, no germ line mutation of TP53 was identified, while two single-nucleotide polymorphisms were identified in four patients. In our area, in which a high rate of familial aggregation was demonstrated, the lack of germ line mutation of TP53 together with the infrequency of mutation of E-cadherin gene seem to limit the role of genetic predisposition in the development of gastric cancer. PMID- 19674072 TI - Breast cancer surgery: an historical narrative. Part III. From the sunset of the 19th to the dawn of the 21st century. AB - The 20th century is marked by significant advances regarding the management of breast cancer. A clear trend towards less aggressive surgical operation was constantly noted. Modified radical mastectomy gradually replaced radical mastectomy during the second half of the 20th century, while during the last two decades breast-conservation therapy became the treatment of choice for the treatment of breast cancer. This type of therapy includes segmental mastectomy (either quadrantectomy or lumpectomy) with axillary lymph node dissection, followed by postoperative irradiation. Other significant advances during the 20th century include the introduction of systemic therapy (chemotherapy, hormonal therapy) and radiation therapy. Better patient follow-up, statistical analysis, development of staging systems and the introduction of frozen section, the development and wide use of mammography (including screening mammography), breast reconstruction following mastectomy and the development of newer diagnostic methods [including breast magnetic resonance imaging and the advanced breast biopsy instrumentation (ABBI)] are other advances that contributed to a better management of breast cancer patients. Sentinel lymph node biopsy has been introduced during the 1990 s in an attempt to reduce morbidity due to axillary lymph node dissection. Despite these advances, breast cancer remains a significant problem and represents a field of active and intense research. PMID- 19674073 TI - Breast cancer surgery: an historical narrative. Part II. 18th and 19th centuries. AB - During the 18th and 19th centuries, management of breast cancer was greatly improved. The humoral theory of Galen, which dominated for centuries, was fallen into disfavour. Axillary nodal involvement was recognised as an adverse prognostic factor, while LeDran, in the middle of the 18th century, proposed the theory of lymphatic spread of breast cancer; he also favoured the idea that breast cancer at its earliest stage was a local disease, which could be effectively treated by surgery. The need to excise enlarged axillary lymph nodes was recognised by other surgeons of the 18th century, including Petit, who proposed a procedure very similar to radical mastectomy. During the 19th century, significant advances were noted, including the development of anaesthesia and antisepsis, a better understanding of the biology of cancer and the introduction of microscopic examination. Radical mastectomy was widely used in clinical practice by Halsted. However, this radical procedure was used by other surgeons of that time, including Meyer. Halsted was able to report a very low local recurrence rates (approximately 6%), a very important achievement given the advanced stages of the breast cancer when diagnosed in women at that time. PMID- 19674074 TI - Breast cancer surgery: an historical narrative. Part I. From prehistoric times to Renaissance. AB - Cancer was known as a disease since prehistoric times. Management of breast cancer evolved slowly through centuries in the ancient world up to the Renaissance. This period is marked by the absence of any scientifically verifiable understanding of the true nature of cancer and its natural history and consequently by a lack of effective treatment. Breast has been considered as a symbol of femininity, fertility and beauty. Hippocrates proposed that breast cancer, among other neoplasms, was a 'systemic disease' caused by an excess of black bile. The humoral theory was further supported by Galen and dominated for centuries in medicine. Fulguration and breast amputation by using various instruments to achieve a rapid operation were widely used up to the 18th century. The Renaissance was a revolutionary period, since it stimulated medical practice; at that time physicians started to scientifically study medicine. Vesalius greatly contributed in the advancement of surgery, and he vigorously opposed Galen's doctrines. Many great surgeons of that time (including Pare, Cabrol, Servetto, Scultetus, Tulp, Fabry von Hilded, etc.) advanced the science of surgery. Interestingly, Bartoleny Gabrol (1590) in Montpellier advocated radical mastectomy, which was popularised by Halsted, 300 years later. However, the lack of anaesthesia and the problem of wound infections (due to the lack of the aseptic techniques) generated significance and often problems for the surgeons of that time. Surgery was often 'heroic' but primitive and even inhumane by current standards. Therapeutic nihilism was the prevailing altitude regarding breast cancer, at least among the vast majority of surgeons. PMID- 19674075 TI - Tracing the incidence of paclitaxel-induced peripheral neuropathy. PMID- 19674076 TI - CXCL16 in HIV infection - a link between inflammation and viral replication. AB - BACKGROUND: While some chemokines are thought to be protective in HIV-infected individuals by their ability to block HIV entry into T cells and macrophages, chemokines could also have harmful effects in HIV infection through their ability to promote inflammation. Here, we examined the regulation and the effects of CXCL16, a newly discovered chemokine of the CXC family, in HIV-infected patients. MATERIALS AND METHODS: We examined serum levels of CXCL16 in clinically well defined subgroups of HIV-infected individuals both before (n = 62) and during HAART (n = 40) as well as in age- and sex-matched healthy controls (n = 30). We also examined the effects of CXCL16 on inflammatory and anti-inflammatory cytokines and HIV replication in peripheral blood mononuclear cells (PBMC). RESULTS: Our main and novel findings were: (i) HIV-infected patients had significant raised CXCL16 levels according to disease severity and progression. (ii) During HAART, the immunological improvement was accompanied by a modest increase in CXCL16 level. (iii) While soluble CXCL16 promoted an anti inflammatory response in PBMC from those on successful HAART, it induced an inflammatory response and enhanced HIV replication in PBMC from those with high viral load irrespectively of ongoing HAART. (iv) Recombinant HIV-tat protein significantly increased CXCL16 release in THP-1 macrophages. CONCLUSIONS: Our findings suggest a complex interaction between CXCL16 and HIV, promoting both inflammatory and anti-inflammatory effects as well as HIV replication, partly dependent on accompanying HIV replication. PMID- 19674077 TI - Erosion of telomeric single-stranded overhang in patients with aplastic anaemia carrying telomerase complex mutations. AB - BACKGROUND: Loss-of-function mutations in telomerase complex genes reduce telomerase activity and shorten overall telomere length in leucocytes, and they can clinically manifest as bone marrow failure (aplastic anaemia and dyskeratosis congenita) and familial pulmonary fibrosis. Telomeres are constituted of double stranded tandem TTAGGG repeats followed by a 3' G-rich single-stranded overhang, a crucial telomeric structural component responsible for the t-loop formation. MATERIALS AND METHODS: We investigated the length of telomeric overhangs in 25 healthy individuals from 0 to 76 years of age, 16 patients with aplastic anaemia, and 13 immediate relatives using a non-denaturing in-gel method and the telomere oligonucleotide ligation assay. RESULTS: Telomeric overhang lengths were constant from birth to eighth decade of life in healthy subjects, in contrast to overall telomere length, which shortened with ageing. Most patients with marrow failure and a telomerase gene mutation showed marked erosion of telomeric overhang associated with critically short telomeres; in other aplastic patients with normal genotypes, normal overall telomere lengths and who responded to immunosuppressive therapy, telomeric overhangs were maintained. CONCLUSIONS: Telomeric overhang erosion does not participate in physiological ageing but support a role for eroded telomeric overhangs and abnormal telomere structure in pathological shortening of telomeres, especially caused by loss-of-function telomerase mutations. Disrupted telomere structure caused by short telomeric overhangs may contribute to the mechanisms of abnormal haematopoietic compartment senescence and chromosomal instability in human bone marrow failure. PMID- 19674078 TI - Postoperative development of aspirin resistance following coronary artery bypass. AB - BACKGROUND: Aspirin therapy is known to substantially reduce mortality and the rate of ischaemic complications after coronary artery bypass grafting (CABG). Rates of perioperative aspirin resistance cited in the literature are up to 50% and could be influenced by extracorporeal circulation. Thus, aspirin resistance after CABG may have a significant clinical relevance. MATERIALS AND METHODS: In 59 patients undergoing CABG (on-pump, off-pump and combined procedures) aspirin resistance was investigated by arachidonic acid induced platelet aggregometry. Clinical relevance was assessed with 12-month follow up. RESULTS: Two types of resistance were observed: A preoperative resistance (despite oral aspirin or in vitro addition) was present in 29% and a postoperative developing type was seen in 49% resulting in only 22% of patients with a 'normal' reaction to aspirin. If patients were already on oral aspirin at admission, the rate of resistance was significantly reduced. Off-pump surgery or pump-times exceeding 120 min had no significant impact on resistance. During the 12-month follow up (98.3%), there were three deaths (one stroke, one intestinal ischaemia, one mediastinitis after postoperative delirium) in patients with the perioperative resistance and none in other patients (P = 0.345). In none of those patients who presented with perioperative aspirin resistance, could this aspirin resistance be demonstrated when tested again after 12 months? CONCLUSIONS: Aspirin resistance is a transient phenomenon present in the majority of patients undergoing CABG. The three deaths in the resistant group may - although not statistically significant - indicate the possibility of a worse outcome for patients with aspirin resistance. PMID- 19674079 TI - Anti-oxidized LDL antibody levels are reduced in women with hypertension. AB - BACKGROUND: Previous studies have suggested that hypertension may be associated with increased oxidized low-density lipoprotein (LDL). Increased in vitro oxidizability of LDL or elevated titers of anti-oxidized LDL antibodies have been shown in subjects with essential hypertension. However, the relationship between oxidized LDL and hypertension is equivocal. We examined the association between hypertension and levels of IgG anti-oxidized LDL antibodies in a group of women from the general population. MATERIALS AND METHODS: The study included 619 women classified according to their blood pressure values. IgG anti-oxidized LDL antibodies were measured by enzyme-linked immunosorbent assay and the women were classified as being above or below the 50th percentile. RESULTS: Hypertension was present in 54.3% of the women. These women had significantly lower levels of IgG anti-oxidized LDL antibodies than the normotensive women (0.280 +/- 0.117 vs. 0.336 +/- 0.125, P < 0.001). Both systolic and the diastolic blood pressures showed a significant negative correlation with the levels of IgG anti-oxidized LDL antibodies (r = -0.204, P < 0.001; r = -0.225, P < 0.001, respectively). Women with IgG anti-oxidized LDL antibody levels above the 50th percentile had a lower prevalence of hypertension than those with IgG anti-oxidized LDL antibody levels below the 50th percentile (40.2% vs. 59.8%) (P < 0.001). CONCLUSIONS: Women with hypertension had lower levels of IgG anti-oxidized LDL antibodies than normotensive women. PMID- 19674080 TI - Demographic and ADAMTS13 biomarker data as predictors of early recurrences of idiopathic thrombotic thrombocytopenic purpura. AB - OBJECTIVE: Approximately 40% of idiopathic thrombotic thrombocytopenic purpura (TTP) patients will suffer an exacerbation (recurrence of TTP within 30 d after their last plasma exchange (PE) procedure), but there are no data to predict who is at greater risk. We studied the clinical utility of demographic and ADAMTS13 biomarker data to predict the risk for exacerbation. PATIENTS: Forty-four acute episodes of idiopathic TTP from 26 patients were studied. METHODS: PE was performed plus either prednisone (1 mg/kg/d) or cyclosporin (2-3 mg/kg/d) as adjuncts. PE was continued daily until response (platelet count >150 000/microL and normalized lactate dehydrogenase) and tapered uniformly in all patients. ADAMTS13 biomarkers were studied prior to PE and after achieving a response, but within 7 d of the last PE. RESULTS: African American race (AA) was associated with an increased risk for exacerbation (P = 0.046). ADAMTS13 at presentation was also significantly lower in patients experiencing an exacerbation (P = 0.0364). After adjusting for the race effect, ADAMTS13 remained marginally significant (P = 0.0569). CONCLUSIONS: AA is significantly associated with an increased risk for exacerbations of TTP. These data also suggest that decreasing pretreatment ADAMTS13 activity was associated with an increased risk for exacerbation, even after accounting for the effect of race. PMID- 19674081 TI - Homeostatic plasticity in a reward processing region: accumbens neurons scale too! (Commentary on Sun & Wolf). PMID- 19674082 TI - Reconstruction of the nigrostriatal dopamine pathway in the adult mouse brain. AB - Transplants of fetal dopamine neurons can be used to restore dopamine neurotransmission in animal models of Parkinson's disease, as well as in patients with advanced Parkinson's disease. In these studies the cells are placed in the striatum rather than in the substantia nigra where they normally reside, which may limit their ability to achieve full restoration of motor function. Using a microtransplantation approach, which allows precise placement of small cell deposits directly into the host substantia nigra, and fetal donor cells that express green fluorescent protein under the control of the tyrosine hydroxylase promoter, we show that dopamine neuroblasts implanted into the substantia nigra of adult mice are capable of generating a new nigrostriatal pathway with an outgrowth pattern that matches the anatomy of the intrinsic system. This target directed regrowth was closely aligned with the intrinsic striatonigral fibre projection and further enhanced by over-expression of glial cell line-derived neurotrophic factor in the striatal target. Results from testing of amphetamine induced rotational behaviour suggest, moreover, that dopamine neurons implanted into the substantia nigra are also capable of integrating into the host circuitry at the functional level. PMID- 19674084 TI - The fusiform face area and occipital face area show sensitivity to spatial relations in faces. AB - Behavioral research indicates that successful face individuation is associated with sensitivity to subtle spatial relations between facial features, as well as to the features themselves. We used a blocked functional magnetic resonance adaptation paradigm to examine the sensitivity of the core face network to spatial relations in faces. The fusiform face area (FFA) was sensitive to spatial relations, responding more strongly to a single face presented with various feature spacings than to repeated presentations of an identical face. This response to spacing variations was as strong as the response to a series of distinct identities. There were no hemisphere effects in sensitivity to spatial relations, although FFAs were larger on the right. The right occipital face area (OFA) was also sensitive to spatial relations in faces. Few participants showed left OFAs. The superior temporal sulcus (STS), which does not code identity, showed little sensitivity to either relational changes or changes in identity. We suggest that the sensitivity of the FFA and right OFA to spatial relations in faces may contribute to our impressive ability to individuate faces despite their similarity as visual patterns. PMID- 19674083 TI - Development of epileptiform excitability in the deep entorhinal cortex after status epilepticus. AB - Epileptiform neuronal activity during seizures is observed in many brain areas, but its origins following status epilepticus (SE) are unclear. We have used the Li low-dose pilocarpine rat model of temporal lobe epilepsy to examine early development of epileptiform activity in the deep entorhinal cortex (EC). We show that during the 3-week latent period that follows SE, an increasing percentage of neurons in EC layer 5 respond to a single synaptic stimulus with polysynaptic burst depolarizations. This change is paralleled by a progressive depolarizing shift of the inhibitory postsynaptic potential reversal potential in layer 5 neurons, apparently caused by upregulation of the Cl(-) inward transporter NKCC1 and concurrent downregulation of the Cl(-) outward transporter KCC2, both changes favoring intracellular Cl(-) accumulation. Inhibiting Cl(-) uptake in the latent period restored more negative GABAergic reversal potentials and eliminated polysynaptic bursts. The changes in the Cl(-) transporters were highly specific to the deep EC. They did not occur in layers 1-3, perirhinal cortex, subiculum or dentate gyrus during this period. We propose that the changes in Cl(-) homeostasis facilitate hyperexcitability in the deep entorhinal cortex leading to epileptiform discharge there, which subsequently affects downstream cortical regions. PMID- 19674085 TI - Behaviorally evoked transient reorganization of hippocampal spines. AB - Dendritic spines, microstructures that receive the majority of excitatory synaptic inputs, are fundamental units to integrate and store neuronal information. The morphological reorganization of spines accompanies the functional alterations in synaptic strength underlying memory-relevant modifications of network connectivity. Here we report the rapid dynamics of cell population-selective spine reorganizations related to behavioral experiences. In Thy1-GFP transgenic mice, hippocampal CA1 pyramidal neurons that were putatively activated during environmental explorations were detected with their post hoc immunoreactivity for Arc, an activity-dependent immediately-early gene. Immediately after a 60-min exposure to a familiar environment, the spine densities of Arc-positive and Arc-negative neurons were differently distributed. This density imbalance was due exclusively to changes in the number of small, rather than large, spines. The change disappeared within 60 min after mice were returned to the home cages. Thus, spines possess the ability to rapidly and reversibly alter their morphology in response to a brief environmental change. We propose that these transient spine dynamics represent a latent preliminary stage for longer-term plasticity on demand. PMID- 19674086 TI - Visual paired-pulse stimulation reveals enhanced visual cortex excitability in migraineurs. AB - Migraine is a common ictal disorder with an interindividual heterogeneous characteristic, whose underlying mechanisms remain elusive. On the one hand migraine is associated with abnormal cortical hyperexcitability. On the other hand, studies reported lower amplitudes of visual-evoked potentials (VEPs) and concluded that low preactivation levels imply decreased excitability. Here we measured visual cortex excitability and paired-pulse suppression in subjects suffering from migraine without aura and in a group of aged- and gender-matched healthy subjects to address the relation between activation levels and excitability. To that aim, we analysed amplitudes of VEPs and paired-pulse suppression evoked by a paired-pulse stimulation paradigm using stimulus onset asynchronies (SOAs) between 80 and 133 ms. We found that in migraineurs in the interictal state the amplitudes of the first VEP were reduced as compared with healthy subjects by approximately 20%. In the case of paired-pulse suppression comparable to healthy controls, the second response amplitude should be reduced as well, which was not the case. Instead, the ratio between the first and second VEP was higher than in healthy controls and did not depend on SOA in the range tested, which demonstrates reduced paired-pulse suppression and therefore implicates increased cortical excitability. Our data show that in migraineurs VEPs were reduced presumably due to reduced activation levels. However, paired pulse suppression using short SOAs in the range of 100 ms or less was even higher than in normal subjects. Thus, our data show that signatures of both hyper- and hypoexcitability can be found depending on stimulation condition. PMID- 19674087 TI - Enhanced dense core granule function and adrenal hypersecretion in a mouse model of Rett syndrome. AB - Rett syndrome (RTT) is a progressive developmental disorder resulting from loss of-function mutations in the gene encoding methyl-CpG-binding protein 2 (MeCP2), a transcription regulatory protein. The RTT phenotype is complex and includes severe cardiorespiratory abnormalities, dysautonomia and behavioral symptoms of elevated stress. These findings have been attributed to an apparent hyperactivity of the sympathetic nervous system due to defects in brainstem development; however, the possibility that the peripheral sympathoadrenal axis itself is abnormal has not been explored. The present study demonstrates that the adrenal medulla and sympathetic ganglia of Mecp2 null mice exhibit markedly reduced catecholamine content compared with wild-type controls. Despite this, null animals exhibit significantly higher plasma epinephrine levels, suggesting enhanced secretory granule function in adrenal chromaffin cells. Indeed, we find that Mecp2 null chromaffin cells exhibit a cell autonomous hypersecretory phenotype characterized by significant increases in the speed and size of individual secretory granule fusion events in response to electrical stimulation. These findings appear to indicate accelerated formation and enhanced dilation of the secretory granule fusion pore, resulting in elevated catecholamine release. Our data therefore highlight abnormal catecholamine function in the sympathoadrenal axis as a potential source of autonomic dysfunction in RTT. These findings may help to explain the apparent 'overactivity' of the sympathetic nervous system reported in patients with RTT. PMID- 19674089 TI - GABA release from cerebellar stellate cells is developmentally regulated by presynaptic GABA(B) receptors in a target-cell-specific manner. AB - Transmitter release from boutons along a common axon is often regulated depending on the postsynaptic target. Here, GABA release from cerebellar stellate cells onto Purkinje cells and other stellate cells was examined in acute cerebellar slices of 2- and 4-week-old mice. Consistent with previous findings on action potential-dependent GABA release, we found a developmental decrease in inhibitory inputs onto Purkinje cells but not onto stellate cells when recording miniature inhibitory postsynaptic currents (mIPSCs). Although amplitudes of mIPSCs were developmentally reduced in both cell types, mIPSC frequencies were decreased in Purkinje cells but were increased in stellate cells. Similarly, modulation of GABA release by presynaptic GABA(B) receptors changed during development in Purkinje cells but not in stellate cells, as demonstrated by the baclofen mediated depression of mIPSC frequency and evoked IPSC (eIPSC) amplitudes. The selectively diminished baclofen effect in 4-week-old Purkinje cells correlated with a selective downregulation of presynaptic GABA(B) receptors at stellate cell to-Purkinje cell synapses observed by immunoelectron microscopy analysis. Thus, expression of GABA(B) receptors in stellate cell axons and presynaptic modulation of GABA release appear to change during development in a target-cell-specific manner. PMID- 19674088 TI - 3D surface perception from motion involves a temporal-parietal network. AB - Previous research has suggested that three-dimensional (3D) structure-from-motion (SFM) perception in humans involves several motion-sensitive occipital and parietal brain areas. By contrast, SFM perception in nonhuman primates seems to involve the temporal lobe including areas MT, MST and FST. The present functional magnetic resonance imaging study compared several motion-sensitive regions of interest including the superior temporal sulcus (STS) while human observers viewed horizontally moving dots that defined either a 3D corrugated surface or a 3D random volume. Low-level stimulus features such as dot density and velocity vectors as well as attention were tightly controlled. Consistent with previous research we found that 3D corrugated surfaces elicited stronger responses than random motion in occipital and parietal brain areas including area V3A, the ventral and dorsal intraparietal sulcus, the lateral occipital sulcus and the fusiform gyrus. Additionally, 3D corrugated surfaces elicited stronger activity in area MT and the STS but not in area MST. Brain activity in the STS but not in area MT correlated with interindividual differences in 3D surface perception. Our findings suggest that area MT is involved in the analysis of optic flow patterns such as speed gradients and that the STS in humans plays a greater role in the analysis of 3D SFM than previously thought. PMID- 19674090 TI - Mechanisms of after-hyperpolarization following activation of fly visual motion sensitive neurons. AB - In many neurons, strong excitatory stimulation causes an after-hyperpolarization (AHP) at stimulus offset, which might give rise to activity-dependent adaptation. Graded-potential visual motion-sensitive neurons of the fly Calliphora vicina respond with depolarization and hyperpolarization during motion in their preferred direction and their anti-preferred direction, respectively. A prominent after-response, opposite in sign to the response during motion, is selectively expressed after stimulation with preferred-direction motion. Previous findings suggested that this AHP is generated in the motion-sensitive neurons themselves rather than in presynaptic processing layers. However, it remained unknown whether the AHP is caused by membrane depolarization itself or by another process, e.g. a signaling cascade triggered by activity of excitatory input channels. Here we showed by current injections and voltage clamp that the AHP and a corresponding current are generated directly by depolarization. To test whether the generation of an AHP is linked to depolarization via a Ca(2+)-dependent mechanism, we used photoactivation of a high-affinity Ca(2+) buffer. In accordance with previous findings the AHP was insensitive to manipulation of cytosolic Ca(2+). We propose that membrane depolarization presents a more direction-selective mechanism for the control of AHP than other potential control parameters. PMID- 19674092 TI - A test of the neutral model of expression change in natural populations of house mouse subspecies. AB - Changes in expression of genes are thought to contribute significantly to evolutionary divergence. To study the relative role of selection and neutrality in shaping expression changes, we analyzed 24 genes in three different tissues of the house mouse (Mus musculus). Samples from two natural populations of the subspecies M. m. domesticus and M. m. musculus were investigated using quantitative PCR assays and sequencing of the upstream region. We have developed an approach to quantify expression polymorphism within such populations and to disentangle technical from biological variation in the data. We found a correlation between expression polymorphism within populations and divergence between populations. Furthermore, we found a correlation between expression polymorphism and sequence polymorphism of the respective genes. These data are most easily interpreted within a framework of a predominantly neutral model of gene expression change, where only a fraction of the changes may have been driven by positive selection. Although most genes investigated were expressed in all three tissues analyzed, significant changes of expression levels occurred predominantly in a single tissue only. This adds to the notion that enhancer specific effects or transregulatory effects can modulate the evolution of gene expression in a tissue-specific way. PMID- 19674093 TI - Rogers' paradox recast and resolved: population structure and the evolution of social learning strategies. AB - We explore the evolution of reliance on social and asocial learning using a spatially explicit stochastic model. Our analysis considers the relative merits of four evolved strategies, two pure strategies (asocial and social learning) and two conditional strategies (the "critical social learner," which learns asocially only when copying fails, and the "conditional social learner," which copies only when asocial learning fails). We find that spatial structure generates outcomes that do not always conform to the finding of earlier theoretical analyses that social learning does not enhance average individual fitness at equilibrium (Rogers' paradox). Although we describe circumstances under which the strategy of pure social learning increases the average fitness of individuals, we find that spatial structure introduces a new paradox, which is that social learning can spread even when it decreases the average fitness of individuals below that of asocial learners. We also show that the critical social learner and conditional social learner both provide solutions to the aforementioned paradoxes, although we find some conditions in which pure (random) social learning out-competes both conditional strategies. Finally, we consider the relative merits of critical and conditional social learning under various conditions. PMID- 19674091 TI - Nucleus accumbens neurons exhibit synaptic scaling that is occluded by repeated dopamine pre-exposure. AB - Synaptic scaling has been proposed as a form of plasticity that may contribute to drug addiction but it has not been previously demonstrated in the nucleus accumbens (NAc), a critical region for addiction. Here we demonstrate bidirectional synaptic scaling in postnatal rat NAc neurons that were co-cultured with prefrontal cortical neurons to restore excitatory input. Prolonged activity blockade (1-3 days) with an AMPA receptor antagonist increased cell surface (synaptic and extrasynaptic) glutamate receptor 1 (GluR1) and GluR2 but not GluR3, as well as GluR1/2 co-localization on the cell surface and total GluR1 and GluR2 protein levels. A prolonged increase in activity (bicuculline, 48 h) produced opposite effects. These results suggest that GluR1/2-containing AMPA receptors undergo synaptic scaling in NAc neurons. GluR1 and GluR2 surface expression was also increased by tetrodotoxin alone or in combination with an N methyl-d-aspartate receptor or AMPA receptor antagonist but not by the l-type Ca(2+) channel antagonist nifedipine. A cobalt-quenching assay confirmed the immunocytochemical results indicating that synaptic scaling after activity blockade did not involve a change in abundance of GluR2-lacking AMPA receptors. Increased AMPA receptor surface expression after activity blockade required protein synthesis and was occluded by inhibition of the ubiquitin-proteasome system. Repeated dopamine (DA) treatment, which leads to upregulation of surface GluR1 and GluR2, occluded activity blockade-induced synaptic scaling. These latter results indicate an interaction between cellular mechanisms involved in synaptic scaling and adaptive mechanisms triggered by repeated DA receptor stimulation, suggesting that synaptic scaling may not function normally after exposure to DA-releasing drugs such as cocaine. PMID- 19674094 TI - Good genes drive female choice for mating partners in the lek-breeding European treefrog. AB - Investigating the mechanisms underlying female mate choice is important for sexual-selection theory, but also for population-genetic studies, because distinctive breeding strategies affect differently the dynamics of gene diversity within populations. Using field-monitoring, genetic-assignment, and laboratory rearing methods, we investigated chorus attendance, mating success and offspring fitness in a population of lek-breeding tree-frogs (Hyla arborea) to test whether female choice is driven by good genes or complementary genes. Chorus attendance explained approximately 50% of the variance in male mating success, but did not correlate with offspring fitness. By contrast, offspring body mass and growth rate correlated with male attractiveness, measured as the number of matings obtained per night of calling. Genetic similarity between mating partners did not depart from random, and did not affect offspring fitness. We conclude that females are able to choose good partners under natural settings and obtain benefits from the good genes, rather than compatible genes, their offspring inherit. This heritability of fitness is likely to reduce effective population sizes below values previously estimated. PMID- 19674095 TI - Simulating range expansion: male species recognition and loss of premating isolation in damselflies. AB - Prolonged periods of allopatry might result in loss of the ability to discriminate against other formerly sympatric species, and can lead to heterospecific matings and hybridization upon secondary contact. Loss of premating isolation during prolonged allopatry can operate in the opposite direction of reinforcement, but has until now been little explored. We investigated how premating isolation between two closely related damselfly species, Calopteryx splendens and C. virgo, might be affected by the expected future northward range expansion of C. splendens into the allopatric zone of C. virgo in northern Scandinavia. We simulated the expected secondary contact by presenting C. splendens females to C. virgo males in the northern allopatric populations in Finland. Premating isolation toward C. splendens in northern allopatric populations was compared to sympatric populations in southern Finland and southern Sweden. Male courtship responses of C. virgo toward conspecific females showed limited geographic variation, however, courtship attempts toward heterospecific C. splendens females increased significantly from sympatry to allopatry. Our results suggest that allopatric C. virgo males have partly lost their ability to discriminate against heterospecific females. Reduced premating isolation in allopatry might lead to increased heterospecific matings between taxa that are currently expanding and shifting their ranges in response to climate change. PMID- 19674096 TI - Environmental viscosity does not affect the evolution of cooperation during experimental evolution of colicigenic bacteria. AB - Cooperation should be favored under environmental conditions allowing the preferential interaction of cooperators among themselves and limiting interactions with defectors. Bacteria cooperating to kill competitors by secreting a toxin evolved during several hundred generations in two environments: a viscous environment that should promote cooperator assortment, and a nonviscous environment that should not allow such preferential interaction. A quantitative decrease in cooperation was observed in all populations, but as expected, cooperation was maintained at a higher level in the viscous environment. Mutants that are resistant against but not producing the toxin were identified at a low frequency in a few populations from the viscous environment and at a high frequency in all the populations from the nonviscous environment. The underlying mutations were identified. Relative fitness of the cooperator and mutant genotypes were obtained with bacteria that were isogenic, except for the identified mutations. Competition experiments indicated that cooperation is not favored by environmental viscosity as imposed in our system and suggested that when it comes to cooperation, environmental viscosity should be considered not only in terms of individual movement, but also in terms of the distribution of the public good. PMID- 19674097 TI - Optimal defensive coloration strategies during the growth period of prey. AB - Defensive coloration that reduces the risk of predation is considered to be widespread in animals. Many closely related species adopt differing coloration strategies during the life cycle, including crypsis, conspicuousness, and ontogenic change between the two coloration types. Here, we use a dynamic state dependent approach to use ecological and intrinsic factors to predict the proportion of the developmental period of immature animals that should be spent as cryptic or conspicuous, and when conspicuous coloration should be reliably associated with investment in defenses. The model predicts that animals should change color more than once during development only in specific circumstances. In contrast, change from crypsis to conspicuous can occur over a range of conditions related to the frequency of detection by predators, but may also depend on the opportunity costs of crypsis and the effect of size on the deterrent effect of conspicuous coloration. We also report the results of a survey of coloration strategies in lepidopteron larvae, and note a qualitative agreement with the predictions of our model in the relationship between body size and coloration strategy. Our results provide explanations for several widespread antipredator coloration phenomena in prey animals, and provide a comprehensive predictive framework for the types of coloration strategies that are employed in nature. PMID- 19674098 TI - Evolution of mimicry patterns in Metriorrhynchus (Coleoptera: Lycidae): the history of dispersal and speciation in southeast Asia. AB - The concept of Mullerian mimicry suggests convergent evolution to an intermediate pattern and does not predict polymorphism in mimicry rings. We examined the evolution of mimicry patterns and the order of divergence of various factors, including the role of aposematic patterns in speciation, in a clade of net-winged beetles with a robust phylogeny that suggests that they dispersed from the Australian to Asian plate. We found strong evidence for the evolution of mimicry via advergence in Metriorrhynchus because older patterns are represented in the Oriental region within more than 100 species of lycids from several lineages. Advergence was likely the cause of the observed intraspecific polymorphism in contrast to the predicted universal monomorphism. Polymorphism was found in populations of two species in Sumatra and Borneo and in populations fine-tuned to subtle variants in various habitats. The advergence is likely to be based on the small population sizes of immigrants. The differences in population sizes result in much higher benefits for dispersing species than native populations. Speciation was trigged by the divergence in aposematic coloration, and the genetic differences accumulated slowly during incomplete isolation. We assumed that the differentiation in genitalia through sexual selection ultimately reinforced speciation initiated by the shift between mimicry patterns. PMID- 19674099 TI - Speciation due to hybrid necrosis in plant-pathogen models. AB - We develop a model for speciation due to postzygotic incompatibility generated by autoimmune reactions. The model is based on frequency-dependent interactions between host plants and their pathogens, which can generate disruptive selection and give rise to speciation if distant phenotypes become reproductively isolated. Based on recent experimental evidence from Arabidopsis, we assume that at the molecular level, incompatibility between host strains is caused by epistatic interactions between two proteins in the plant immune system--the guard and the guardee. Within each plant strain, immune reactions occur when the guardee protein is modified by a pathogen effector, and the guard subsequently binds to the guardee, thus precipitating an immune response. When guard and guardee proteins come from phenotypically distant parents, a hybrid's immune system can be triggered by erroneous interactions between these proteins even in the absence of pathogen attack, leading to severe autoimmune reactions in hybrids. This generates a Dobzhnasky-Muller incompatibility due to immune reactions. Our model shows how phenotypic variation generated by frequency-dependent host-pathogen interactions can lead to such postzygotic incompatibilities between extremal types, and hence to speciation. PMID- 19674100 TI - Recombinant expression of an insulin-like peptide 3 (INSL3) precursor and its enzymatic conversion to mature human INSL3. AB - Insulin-like peptide 3 (INSL3), which is primarily expressed in the Leydig cells of the testes, is a member of the insulin superfamily of peptide hormones. One of its primary functions is to initiate and mediate descent of the testes of the male fetus via interaction with its G protein-coupled receptor, RXFP2. Study of the peptide has relied upon chemical synthesis of the separate A- and B-chains and subsequent chain recombination. To establish an alternative approach to the preparation of human INSL3, we designed and recombinantly expressed a single chain INSL3 precursor in Escherichia coli cells. The precursor was solubilized from the inclusion body, purified almost to homogeneity by immobilized metal-ion affinity chromatography and refolded efficiently in vitro. The refolded precursor was subsequently converted to mature human INSL3 by sequential endoproteinase Lys C and carboxypeptidase B treatment. CD spectroscopic analysis and peptide mapping showed that the refolded INSL3 possessed an insulin-like fold with the expected disulfide linkages. Recombinant human INSL3 demonstrated full activity in stimulating cAMP activity in RXFP2-expressing cells. Interestingly, the activity of the single-chain precursor was comparable with that of the mature two-chain INSL3, suggesting that the receptor-binding region within the mid- to C-terminal of B-chain is maintained in an active conformation in the precursor. This study not only provides an efficient approach for mature INSL3 preparation, but also resulted in the acquisition of a useful single-chain template for additional structural and functional studies of the peptide. PMID- 19674101 TI - A novel, promoter-based, target-specific assay identifies 2-deoxy-D-glucose as an inhibitor of globotriaosylceramide biosynthesis. AB - Abnormal biosynthesis of globotriaosylceramide (Gb3) is known to be associated with Gb3-related diseases, such as Fabry disease. The Gb3 synthase gene (Gb3S) codes for alpha1,4-galactosyltransferase, which is a key enzyme involved in Gb3 biosynthesis in vivo. Transcriptional repression of Gb3S is a way to control Gb3 biosynthesis and may be a suitable target for the treatment of Gb3-related diseases. To find a transcriptional inhibitor for Gb3S, we developed a convenient cell-based chemical screening assay system by constructing a fusion gene construct of the human Gb3S promoter and a secreted luciferase as reporter. Using this assay, we identified 2-deoxy-D-glucose as a potent inhibitor for the Gb3S promoter. In cultured cells, 2-deoxy-D-glucose markedly reduced endogenous Gb3S mRNA levels, resulting in a reduction in cellular Gb3 content and a corresponding accumulation of the precursor lactosylceramide. Moreover, cytokine-induced expression of Gb3 on the cell surface of endothelial cells, which is closely related to the onset of hemolytic uremic syndrome in O157-infected patients, was also suppressed by 2-deoxy-D-glucose treatment. These results indicate that 2 deoxy-D-glucose can control Gb3 biosynthesis through the inhibition of Gb3S transcription. Furthermore, we demonstrated the general utility of our novel screening assay for the identification of new inhibitors of glycosphingolipid biosynthesis. PMID- 19674102 TI - Conformational stability of neuroglobin helix F--possible effects on the folding pathway within the globin family. AB - Neuroglobin is a recently discovered member of the globin family, mainly observed in neurons and retina. Despite the low sequence identity (less than 20% over the whole sequence for the human proteins), the general fold of neuroglobin closely resembles that of myoglobin. The latter is a paradigmatic protein for folding studies, whereas much less is known about the neuroglobin folding pathway. In this work, we show how the structural features of helix F in neuroglobin and myoglobin could represent a pivotal difference in their folding pathways. Former studies widely documented that myoglobin lacks helix F in the apo form. In this study, limited proteolysis experiments on aponeuroglobin showed that helix F does not undergo proteolytic cleavage, suggesting that, also in the apo form, this helix maintains a rigid and structured conformation. To understand better the structural properties of helices F in the two proteins, we analyzed peptides encompassing helix F of neuroglobin and myoglobin in the wild-type and mutant forms. NMR and CD experiments revealed a helical conformation for neuroglobin helix F peptide, at both pH 7 and pH 2, absent in the myoglobin peptide. In particular, NMR data suggest a secondary structure stabilization effect caused by hydrophobic interactions involving Tyr88, Leu89 and Leu92. Molecular dynamics simulations performed on the apo and holo forms of the two proteins reveal the persistence of helix F in neuroglobin even in the absence of heme. Conversely myoglobin shows a higher mobility of the N-terminus of helix F on heme removal, which leads to the loss of secondary structure. PMID- 19674103 TI - Changes in microRNAs associated with hepatic stellate cell activation status identify signaling pathways. AB - Activation of hepatic stellate cells (HSCs), which is regulated by multiple signal transduction pathways, is the key event in liver fibrosis. Moreover, members of these pathways are important targets for microRNAs (miRNAs). To better understand the critical pathways of HSC activation, we performed comprehensive comparative bioinformatics analysis of microarrays of quiescent and activated HSCs. Changes in miRNAs associated with HSC activation status revealed that 13 pathways were upregulated and 22 pathways were downregulated by miRNA. Furthermore, mitochondrial integrity, based on highly upregulated Bcl-2 and downregulated caspase-9, was confirmed in HSCs and fibrotic livers by immnofluorescence assay, quantitative RT-PCR, and western blot analysis. These findings provide in vitro and in vivo evidence that the mitochondrial pathway of apoptosis plays a significant role in the progression of liver fibrogenesis via HSC activation. PMID- 19674104 TI - Mutation of epidermal growth factor receptor is associated with MIG6 expression. AB - Controlled activation of epidermal growth factor receptor (EGFR) is systematically guaranteed at the molecular level; however, aberrant activation of EGFR is frequently found in cancer. Transcription induced by EGFR activation often involves the coordinated expression of genes that positively and negatively regulate the original signaling pathway; therefore, alterations in EGFR kinase activity may reflect changes in gene expression associated with the pathway. In the present study, we investigated transcriptional changes after EGF stimulation with or without the EGFR kinase inhibitor Iressa in H1299 human non-small-cell lung cancer cells [parental H1299, H1299 cells that overexpress wild-type EGFR (EGFR-WT) and mutant H1299 cells that overexpress EGFR where Leu858 is substituted with Arg (L858R)]. The results obtained clearly demonstrate differences in transcriptional activity in the absence or presence of EGFR kinase activity, with genes sharing the same molecular functions showing distinct expression dynamics. The results show the particular enrichment of EGFR/ErbB signaling-related genes in a differentially expressed gene set, and significant protein expression of MIG6/RALT(ERRFI1), an EGFR negative regulator, was confirmed in L858R. High MIG6 protein expression was correlated with basal EGFR phosphorylation and inversely correlated with EGF-induced extracellular signal regulated protein kinase phosphorylation levels. Investigation of the NCI-60 cell lines showed that ERRFI1 expression was correlated with EGFR expression, regardless of tissue type. These results suggest that cells accumulate MIG6 as an inherent negative regulator to suppress excess EGFR activity when basal EGFR kinase activity is considerably high. Taking all the above together, an EGFR mutation can cause transcriptional changes to accommodate the activation potency of the original signaling pathway at the cellular level. PMID- 19674105 TI - CAP2 enhances germination of transgenic tobacco seeds at high temperature and promotes heat stress tolerance in yeast. AB - We reported earlier that ectopic expression of CAP2, a single AP2 domain containing transcription activator from chickpea (Cicer arietinum) in tobacco improves growth and development, and tolerance to dehydration and salt stress, of the transgenic plants. Here, we report that, in addition, the CAP2-transgenic tobacco seeds also exhibit higher germination efficiency at high temperature and show higher expression levels of genes for tobacco heat shock proteins and a heat shock factor. CAP2 was able to activate the 5'-upstream activating sequence of tobacco heat shock factor. Surprisingly, expression of CAP2 cDNA in Saccharomyces cerevisiae also enhanced heat tolerance, with increased expression of the gene for yeast heat shock factor 1 (Hsf1) and its target, the gene for yeast heat shock protein 104 (Hsp104). Sequence analysis of the Hsf1 promoter revealed the presence of a dehydration-responsive element/C-repeat-like element (DRE/CRE). Recombinant CAP2 protein bound to the DRE/CRE in the Hsf1 promoter in a gel shift assay and transactivated the Hsf1 promoter-His reporter construct. The full length CAP2 protein was required to provide thermotolerance in yeast. If these findings are taken together, our results suggest that CAP2 is involved in the heat stress response and provides an example of functioning of a plant transcription factor in yeast, highlighting the strong evolutionary conservation of the stress response mechanism. PMID- 19674106 TI - The enzymatic activity of SR protein kinases 1 and 1a is negatively affected by interaction with scaffold attachment factors B1 and 2. AB - SR protein kinases (SRPKs) phosphorylate Ser/Arg dipeptide-containing proteins that play crucial roles in a broad spectrum of basic cellular processes. Phosphorylation by SRPKs constitutes a major way of regulating such cellular mechanisms. In the past, we have shown that SRPK1a interacts with the nuclear matrix protein scaffold attachment factor B1 (SAFB1) via its unique N-terminal domain, which differentiates it from SRPK1. In this study, we show that SAFB1 inhibits the activity of both SRPK1a and SRPK1 in vitro and that its RE-rich region is redundant for the observed inhibition. We demonstrate that kinase activity inhibition is caused by direct binding of SAFB1 to SRPK1a and SRPK1, and we also present evidence for the in vitro binding of SAFB2 to the two kinases, albeit with different affinity. Moreover, we show that both SR protein kinases can form complexes with both scaffold attachment factors B in living cells and that this interaction is capable of inhibiting their activity, depending on the tenacity of the complex formed. Finally, we present data demonstrating that SRPK/SAFB complexes are present in the nucleus of HeLa cells and that the enzymatic activity of the nuclear matrixlocalized SRPK1 is repressed. These results suggest a new role for SAFB proteins as regulators of SRPK activity and underline the importance of the assembly of transient intranuclear complexes in cellular regulation. PMID- 19674108 TI - Substitution of residues at the double dimer interface affects the stability and oligomerization of goose delta-crystallin. AB - Delta-crystallin is the major structural protein in avian and reptilian eye lenses, and confers special refractive properties. The protein is a homotetramer arranged as a dimer of dimers. In the present study, the roles of the side chains of Glu267, Lys315, and Glu327, which provide hydrogen bonds at the double dimer interface, were investigated. Hydrophobic side chain substitution led to all mutant proteins having an unstable dimer interface. The E267L/E327L mutant had the greatest sensitivity to temperature, urea and guanidinium hydrochloride denaturation, and the most extensive exposure of hydrophobic patches, as judged by 1-anilinonaphthalene-8-sulfonic acid fluorescence, CD, and tryptophan fluorescence. In contrast, the E267L/K315L/E327L mutant showed higher stability than the E267L/E327L mutant. Some level of the dissociated dimeric form was observed in the K315L mutant, but it was not observed for the K315A and E267L/K315L mutants. The E327L mutant was partially in the dissociated dimeric form, whereas the E267/E327L mutant was predominantly dissociated into dimers. In contrast, the triple mutant of E267L/K315L/E327L retained a tetrameric structure. In the presence of urea, a stable monomeric intermediate with higher stability than the wild type was identified for the K315A mutant. Disruption of interfacial interactions at Glu267 led to polymerization of partly unfolded intermediates in the presence of 3 m urea. However, these polymeric forms were not observed with combinations of the E267L mutation with other mutations. These results indicate that these hydrogen bonds, which are present at different contact surfaces in the dimer-dimer interface, perform distinct functions in double dimer assembly. The coordination of these interactions is critical for the stability and tetramer formation of delta-crystallin. PMID- 19674107 TI - Substrate specificity and excision kinetics of natural polymorphic variants and phosphomimetic mutants of human 8-oxoguanine-DNA glycosylase. AB - Human 8-oxoguanine-DNA glycosylase (OGG1) efficiently removes mutagenic 8-oxo-7,8 dihydroguanine (8-oxoGua) and 2,6-diamino-4-hydroxy-5-formamidopyrimidine when paired with cytosine in oxidatively damaged DNA. Excision of 8-oxoGua mispaired with adenine may lead to G-->T transversions. Post-translational modifications such as phosphorylation could affect the cellular distribution and enzymatic activity of OGG1. Mutations and polymorphisms of OGG1 may affect the enzymatic activity and have been associated with increased risk of several cancers. In this study, we used double-stranded oligodeoxynucleotides containing 8-oxoGua:Cyt or 8 oxoGua:Ade pairs, as well as gamma-irradiated calf thymus DNA, to investigate the kinetics and substrate specificity of several known OGG1 polymorphic variants and phosphomimetic Ser-->Glu mutants. Among the polymorphic variants, A288V and S326C displayed opposite-base specificity similar to that of wild-type OGG1, whereas OGG1-D322N was 2.3-fold more specific for the correct opposite base than the wild type enzyme. All phosphomimetic mutants displayed approximately 1.5-3-fold lower ability to remove 8-oxoGua in both assays, whereas the substrate specificity of the phosphomimetic mutants was similar to that of the wild-type enzyme. OGG1 S326C efficiently excised 8-oxoGua from oligodeoxynucleotides and 2,6-diamino-4 hydroxy-5-formamidopyrimidine from gamma-irradiated DNA, but excised 8-oxoG rather inefficiently from gamma-irradiated DNA. Otherwise, kcat values for 8 oxoGua excision obtained from both types of experiments were similar for all OGG1 variants studied. It is known that the human AP endonuclease APEX1 can stimulate OGG1 activity by increasing its turnover rate. However, when wild-type OGG1 was replaced by one of the phosphomimetic mutants, very little stimulation of 8 oxoGua removal was observed in the presence of APEX1. PMID- 19674109 TI - Focal localization of MukBEF condensin on the chromosome requires the flexible linker region of MukF. AB - Condensin complexes are the key mediators of chromosome condensation. The MukB MukE-MukF complex is a bacterial condensin, in which the MukB subunit forms a V shaped dimeric structure with two ATPase head domains. MukE and MukF together form a tight complex, which binds to the MukB head via the C-terminal winged helix domain (C-WHD) of MukF. One of the two bound C-WHDs of MukF is forced to detach from two ATP-bound, engaged MukB heads, and this detachment reaction depends on the MukF flexible linker preceding the C-WHD. Whereas MukB is known to focally localize at particular positions in cells by an unknown mechanism, mukE- or mukF-null mutation causes MukB to become dispersed in cells. Here, we report that mutations in MukF causing a defect in the detachment reaction interfere with the focal localization of MukB, and that the dispersed distribution of MukB in cells correlates directly with defects in cell growth and division. The data strongly suggest that the MukB-MukE-MukF condensin forms huge clusters through the ATP-dependent detachment reaction, and this cluster formation is critical for chromosome condensation by this machinery. We also show that the MukF flexible linker is involved in the dimerization and ATPase activity of the MukB head. PMID- 19674110 TI - The beta-N-acetylglucosaminidases NAG1 and NAG2 are essential for growth of Trichoderma atroviride on chitin. AB - The chitinolytic enzyme machinery of fungi consists of chitinases and beta-N acetylglucosaminidases. These enzymes are important during the fungal life cycle for degradation of exogenous chitin, which is the second most abundant biopolymer, as well as fungal cell-wall remodelling. In addition, involvement of chitinolytic enzymes in the lysis of the host cell wall in mycoparasitic Trichoderma spp. has been reported. In view of the fact that fungi have on average 15-20 chitinases, but only two beta-N-acetylglucosaminidases, the question arises how important the latter enzymes actually are for various aspects of chitin degradation. In this study, the role of two beta-N acetylglucosaminidases, NAG1 and NAG2, was analysed in the mycoparasitic fungus Trichoderma atroviride. No beta-N-acetylglucosaminidase activity was detected in T. atrovirideDeltanag1Deltanag2 strains, suggesting that NAG1 and NAG2 are the only enzymes in T. atroviride that possess this activity. Deltanag1Deltanag2 strains were not able to grow on chitin and chitobiose, but the presence of either NAG1 or NAG2 was sufficient to restore growth on chitinous carbon sources in solid media. Our results demonstrated that T. atroviride cannot metabolize chitobiose but only the monomer N-acetylglucosamine, and that N acetylglucosaminidases are therefore essential for the use of chitin as a nutrient source. NAG1 is predominantly secreted into the medium, whereas NAG2 mainly remains attached to the cell wall. No physiological changes or reduction of the mycoparasitic potential of T. atroviride was detected in the double knockout strains, suggesting that the use of chitin as carbon source is only of minor importance for these processes. PMID- 19674111 TI - Cholinoceptive and cholinergic properties of cardiomyocytes involving an amplification mechanism for vagal efferent effects in sparsely innervated ventricular myocardium. AB - Our recent studies have shown that, as indicated by vagal stimulation, an acetylcholinesterase inhibitor donepezil, an anti-Alzheimer's disease drug, prevents progression of heart failure in rats with myocardial infarction, and activates a common cell survival signal shared by acetylcholine (ACh) in vitro. On the basis of this and evidence that vagal innervation is extremely poor in the left ventricle, we assessed the hypothesis that ACh is produced by cardiomyocytes, which promotes its synthesis via a positive feedback mechanism. Rat cardiomyocytes expressed choline acetyltransferase (ChAT) in the cytoplasm and vesicular acetylcholine transporter with the vesicular structure identified by immunogold electron microscopy, suggesting that cardiomyocytes possess components for ACh synthesis. Intracellular ACh in rat cardiomyocytes was identified with physostigmine or donepezil. However, with atropine, the basal ACh content was reduced. In response to exogenous ACh or pilocarpine, cardiomyocytes increased the transcriptional activity of the ChAT gene through a muscarinic receptor and ChAT protein expression, and, finally, the intracellular ACh level was upregulated by pilocarpine. Knockdown of ChAT by small interfering RNA accelerated cellular energy metabolism, which is suppressed by ACh. Although physostigmine had a minimal effect on the ChAT promoter activity by inhibiting acetylcholinesterase, donepezil resulted in elevation of the activity, protein expression and intracellular ACh level even in the presence of sufficient physostigmine. Orally administered donepezil in mice increased the ChAT promoter activity in a reporter gene-transferred quadriceps femoris muscle and the amount of cardiac ChAT protein. These findings suggest that cardiomyocytes possess an ACh synthesis system, which is positively modulated by cholinergic stimuli. Such an amplification system in cardiomyocytes may contribute to the beneficial effects of vagal stimulation on the ventricles. PMID- 19674112 TI - Structure and potential function of gamma-aminobutyrate type A receptor associated protein. AB - The gamma-aminobutyrate type A receptor-associated protein (GABARAP) is a ubiquitin-like modifier, and is implicated in a variety of membrane trafficking and fusion events that are crucial to synaptic plasticity, autophagy and apoptosis. However, important aspects of GABARAP function and regulation remain poorly understood. We review the current state of knowledge about GABARAP, highlighting newly-identified GABARAP ligands, and discuss the possible physiological relevance of each ligand interaction. PMID- 19674113 TI - Cystic fibrosis infections: treatment strategies and prospects. AB - Pseudomonas aeruginosa and Burkholderia cepacia are the two major Gram-negative rods that colonize/infect the lungs of patients with cystic fibrosis (CF). These organisms may cause progressive respiratory failure, although occasionally more rapid infections result in the 'Cepacia' syndrome. Many antibiotics have been used against Pseudomonas and Burkholderia, but once chronic colonization has been established, eradication of these organisms is rare. Drug therapy for CF patients is compromised by a number of bacterial factors that render the infectious agents resistant to antibiotics, including efflux pumps that remove antibiotics, lack of penetration of antibiotics into bacterial biofilms, and changes in the cell envelope that reduce the permeability of antibiotics. Any combination of these mechanisms increases the likelihood of bacterial survival. Therefore, combinations of antibiotics or of antibiotic and nonantibiotic compounds are currently being tested against Pseudomonas and Burkholderia. However, progress has been slow, with only occasional combinations showing promise for the eradication of persistent Gram-negative rods in the airways of CF patients. This review will summarize the current knowledge of CF infections and speculate on potential future pathways to treat these chronic infections. PMID- 19674114 TI - The uropathogenic species Staphylococcus saprophyticus tolerates a high concentration of D-serine. AB - Human urine contains a relatively high concentration of d-serine, which is toxic to several nonuropathogenic bacteria, but can be utilized or detoxified by uropathogenic Escherichia coli (UPEC). The sequenced genome of uropathogenic Staphylococcus saprophyticus contains a gene with homology to the d-serine deaminase gene (dsdA) of UPEC. We found the gene in several clinical isolates of S. saprophyticus; however, the gene was absent in Staphylococcus xylosus and Staphylococcus cohnii, phylogenetically close relatives of S. saprophyticus, and could also not be detected in isolates of Staphylococcus aureus, Staphylococcus epidermidis and 13 other staphylococcal species. In addition, the genomes of other sequenced staphylococci do not harbor homologues of this operon. Interestingly, S. saprophyticus could grow in media supplemented with relatively high concentrations of d-serine, whereas S. aureus, S. epidermidis and other staphylococcal species could not. The association of the dsdA gene with growth in media including d-serine was proved by introducing the gene into S. aureus Newman. Given the fact that UPEC and S. saprophyticus tolerate this compound, d serine utilization and detoxification may be a general property of uropathogenic bacteria. PMID- 19674115 TI - Recurrent dysosmia induced by pyrazinamide. AB - Pyrazinamide can have adverse effects such as hepatic toxicity, hyperuricemia or digestive disorders. In rare cases, alterations in taste and smell function have been reported for pyrazinamide when combined with other drugs. We report a case of reversible olfactory disorder related to pyrazinamide in a woman, with a positive rechallenge. The patient presented every day a sensation of smelling something burning 15 min after drug intake. Dysosmia disappeared completely after pyrazinamide withdrawal and recurred after its rechallenge. The case was reported to the Tunisian Centre of Pharmacovigilance. PMID- 19674116 TI - Molecular aspects of ischaemic postconditioning. AB - Preconditioning, a well established phenomenon had been used since 1980s to attenuate ischaemia-reperfusion induced injury. However, inability to predict the onset of ischaemia in clinical settings led to the discovery of a new concept of postconditioning (PoCo), in 2000s whereby brief repetitive cycles of ischaemia with intermittent reperfusion followed by prolonged ischaemia-elicited tissue protection. There is an impressive array of molecular mechanisms contributing to PoCo-mediated tissue-protection, which include triggers like adenosine (ADO), opioid, erythropoietin (EPO), endogenous nitric-oxide, reactive oxygen species, acetylcholine, tissue factors, pro-inflammatory cytokines and bradykinin; mediators like reperfusion injury salvage kinase pathways including phosphoinositide-3-kinase, extra-cellular signal regulated kinase(1/2) pathway, protein kinase G and protein kinase C; end-effectors like mitochondrial permeability transition pore and mitochondrial potassium ATP channel. The clinical applicability of PoCo has been extended with the use of PoCo mimetic agents like insulin, glucagon like peptide, EPO, statins and ADO before reperfusion in patients with ischaemia reperfusion injury. Remote PoCo has also emerged as a new concept; however, considerable research is required for understanding its molecular mechanisms. In this review, an exhaustive attempt has been made to unearth some molecular aspects of PoCo. PMID- 19674117 TI - Non-adrenergic non-cholinergic inhibition of gastrointestinal smooth muscle and its intracellular mechanism(s). AB - Relaxation of gastrointestinal smooth muscle caused by release of non-adrenergic non-cholinergic (NANC) transmitters from enteric nerves occurs in several physiologic digestive reflexes. Likely candidate NANC inhibitory agents include nitric oxide (NO), adenosine triphosphate (ATP), vasoactive intestinal peptide (VIP), pituitary adenylate cyclase-activating peptide (PACAP), carbon monoxide (CO), protease-activated receptors (PARs), hydrogen sulfide (H2S), neurotensin (NT) and beta-nicotinamide adenine dinucleotide (beta-NAD). Multiple NANC transmitters work in concert, are pharmacologically coupled and are closely coordinated. Individual contribution varies regionally in the gastrointestinal tract and between species. NANC inhibition of gastrointestinal smooth muscle involves several intracellular mechanisms, including increase of cyclic guanosine monophosphate (cGMP), increase of cyclic adenosine monophosphate (cAMP) and hyperpolarization of the cell membrane via direct or indirect activation of potassium ion (K+) channels. PMID- 19674118 TI - Single mother-daughter pair analysis to clarify the diffusion properties of yeast prion Sup35 in guanidine-HCl-treated [PSI] cells. AB - The yeast prion [PSI(+)] is a protein-based heritable element, in which aggregates of Sup35 protein are transmitted to daughter cells in a non-Mendelian manner. To elucidate the mechanism of the transmission, we have developed methods to directly analyse the dynamics of Sup35 fused with GFP in single mother daughter pairs. As it is known that the treatment of yeast cells with guanidine hydrochloride (GuHCl) cures [PSI(+)] by perturbing Hsp104, a prion-remodelling factor, we analysed the diffusion profiles of Sup35-GFP in GuHCl-treated [PSI(+)] cells using fluorescence correlation spectroscopy (FCS). FCS analyses revealed that Sup35-GFP diffusion in the daughter cells was faster; that is, the Sup35-GFP particle was smaller, than that in the mother [PSI(+)] cells, and it eventually reached the diffusion profiles in [psi(-)] cells. We then analysed the flux of Sup35-GFP oligomers from mother to daughter [PSI(+)] cells in the presence of GuHCl, using a modified fluorescent recovery after photobleaching technique, and found that the flux of the diffuse oligomers was completely inhibited. The noninvasive methods described here can be applied to other protein-based transmissible systems inside living cells. PMID- 19674119 TI - R168H and V165X mutant podocin might induce different degrees of podocyte injury via different molecular mechanisms. AB - A lot of mutations of podocin, a key protein of podocyte slit diaphragm (SD), have been found both in hereditary and sporadic focal segmental glomeruloscleorosis (FSGS). Nevertheless, the mechanisms of podocyte injury induced by mutant podocins are still unclear. A compound heterozygous podocin mutation was identified in our FSGS patient, leading to a truncated (podocin (V165X)) and a missense mutant protein (podocin (R168H)), respectively. Here, it was explored whether and how both mutant podocins induce podocyte injury in the in vitro cultured podocyte cell line. Our results showed that podocin (R168H) induced more significant podocyte apoptosis and expression changes in more podocyte molecules than podocin (V165X). Podocyte injury caused by the normal localized podocin(V165X) was effectively inhibited by TRPC6 knockdown. The abnormal retention of podocin(R168H) in endoplasmic reticulum (ER) resulted in the mis-localizations of other critical SD molecules nephrin, CD2AP and TRPC6, and significantly up-regulated ER stress markers Bip/grp78, p-PERK and caspase 12. These results implicated that podocin (R168H) and podocin (V165X) induced different degrees of podocyte injury, which might be resulted from different molecular mechanisms. Our findings provided some possible clues for further exploring the pharmacological targets to the proteinuria induced by different mutant podocins. PMID- 19674120 TI - Impaired Pavlovian fear extinction is a common phenotype across genetic lineages of the 129 inbred mouse strain. AB - Fear extinction is impaired in psychiatric disorders such as post-traumatic stress disorder and schizophrenia, which have a major genetic component. However, the genetic factors underlying individual variability in fear extinction remain to be determined. By comparing a panel of inbred mouse strains, we recently identified a strain, 129S1/SvImJ (129S1), that exhibits a profound and selective deficit in Pavlovian fear extinction, and associated abnormalities in functional activation of a key prefrontal-amygdala circuit, as compared with C57BL/6J. The first aim of the present study was to assess fear extinction across multiple 129 substrains representing the strain's four different genetic lineages (parental, steel, teratoma and contaminated). Results showed that 129P1/ReJ, 129P3/J, 129T2/SvEmsJ and 129X1/SvJ exhibited poor fear extinction, relative to C57BL/6J, while 129S1 showed evidence of fear incubation. On the basis of these results, the second aim was to further characterize the nature and specificity of the extinction phenotype in 129S1, as an exemplar of the 129 substrains. Results showed that the extinction deficit in 129S1 was neither the result of a failure to habituate to a sensitized fear response nor an artifact of a fear response to (unconditioned) tone per se. A stronger conditioning protocol (i.e. five x higher intensity shocks) produced an increase in fear expression in 129S1, relative to C57BL/6J, due to rapid rise in freezing during tone presentation. Taken together, these data show that impaired fear extinction is a phenotypic feature common across 129 substrains, and provide preliminary evidence that impaired fear extinction in 129S1 may reflect a pro-fear incubation-like process. PMID- 19674121 TI - Examination of tetrahydrobiopterin pathway genes in autism. AB - Autism is a complex disorder with a high degree of heritability and significant phenotypic and genotypic heterogeneity. Although candidate gene studies and genome-wide screens have failed to identify major causal loci associated with autism, numerous studies have proposed association with several variations in genes in the dopaminergic and serotonergic pathways. Because tetrahydrobiopterin (BH4) is the essential cofactor in the synthesis of these two neurotransmitters, we genotyped 25 SNPs in nine genes of the BH4 pathway in a total of 403 families. Significant nominal association was detected in the gene for 6-pyruvoyl tetrahydropterin synthase, PTS (chromosome 11), with P = 0.009; this result was not restricted to an affected male-only subset. Multilocus interaction was detected in the BH4 pathway alone, but not across the serotonin, dopamine and BH4 pathways. PMID- 19674123 TI - Snapshot of flexible funding outcomes in four countries. AB - This article reviews social participation outcomes identified in discrete studies of flexible funding programmes across four countries. The outcomes of an Australian flexible funding support programme were studied in 2007; a study tour of independent living programmes was conducted in England and Scotland during 2005; Swedish co-operatives and government administrators providing personal assistance to live independently were visited in 2006 and Australian independent living support groups operating for over 20 years were visited in 2008. Fifty-six interviews were conducted with people with a disability, families, support services, government administrators and researchers. A structured interview schedule was used in the 2007 Australian study and a semi-structured format was used in all other studies. Notes from the interviews were reviewed for themes related to social participation and their contributing factors. Ecological systems theory was used to identify what factors from the micro to the macro system level facilitated or hindered social participation. The key finding is that flexible funding did result in a range of social participation activities in each setting studied. The studies also indicate that social participation increases when people have access to information and support services; can choose their individual workers and move to a new agency if need be; and have adequate resources to meet their needs. The cultural and political context plays a large part in determining these factors. The implications of this study are that adequate resources are needed and the complex systems impacting on flexible funding need to be understood to achieve the intended outcomes. PMID- 19674124 TI - Persuasion and types of enticements offered by pharmaceutical companies to Gold Coast general practitioners in an attempt to encourage a health website recommendation. AB - This study was undertaken to determine if pharmaceutical companies persuade and offer enticements to Gold Coast (Queensland, Australia) general practitioners (GPs), in an attempt to encourage them to recommend health websites to the health consumer. A survey instrument consisting of seven single response questions was mailed to 250 (61%) out of 410 GPs. Questions were designed to measure the percentages (%) and proportions associated with levels of persuasion and types of enticements pharmaceutical companies are offering GPs, in an attempt to encourage them to recommend health websites to the health consumer. The survey instrument allowed participants to indicate their (1) gender, (2) age and (3) years of experience (less experienced or= 10 years). One hundred and eight (43%) of the 250 GPs returned a completed survey. The return rate for male GPs was 72 (40%) and for female GPs, it was 36 (33%). Sixty-eight (63%) GPs indicated that they actively recommend health websites to their patients [male GPs--48 (71%), female GPs--20 (29%)]. This study highlights that female GPs (80%), those aged between 31 and 40 (77%) and GPs with < 10 years experience (72%) were more frequently targeted by pharmaceutical companies. This study reports that pharmaceutical companies are offering various types of enticements in an attempt to persuade Gold Coast GPs to recommend specific health websites to the health consumer. Further research should explore if similar levels of persuasion and types of enticements are being offered to GPs across Australia. PMID- 19674125 TI - A non-randomised controlled trial of the Home Independence Program (HIP): an Australian restorative programme for older home-care clients. AB - The Home Independence Program (HIP) is a short-term restorative programme targeted at older home-care clients, who do not have a diagnosis of dementia, when they are first referred for assistance or when they are referred for additional services because their needs have increased. This study compared the outcomes for individuals who participated in HIP with those of individuals who received 'usual' home-care services. The study was conducted in metropolitan Perth, Western Australia, between 2001 and 2003, when HIP was being trialled as a service in just one region. One hundred clients were recruited into each group and were visited at home on three occasions--service start and at 3 months and 1 year. Standardised outcome measures were used to measure functional dependency, morale, confidence in performing everyday activities without falling and functional mobility. Service outcomes were also examined at 3 months and 1 year. The HIP group showed improvements on all personal outcome measures compared with the control group. These improvements were, except for the morale scale, significantly associated with group assignment even when baseline differences between the groups were adjusted for. As regards service outcomes, the odds of the individuals who received HIP still requiring services was 0.07 (95% CI = 0.03 0.15, P < 0.001) times those for the individuals in the control group at 3 months and 0.14 times at 12 months (95% CI = 0.07-0.29, P < 0.001). The results of this study supported the hypothesis that older individuals referred for home care who participated in a programme to promote their independence had better individual and service outcomes than individuals who received usual home care. PMID- 19674126 TI - Occipital nerve blocks: when and what to inject? AB - INTRODUCTION: Occipital nerve block (ONB) is a promising treatment for headaches. Its indications, selection criteria, and best techniques are not clear, however. OBJECTIVE: To summarize in narrative format what is known about ONBs and what needs to be learned. METHODS: MD Consult and Google Scholar were searched using the terms occipital, suboccipital, block, and injection to identify relevant articles that were reviewed. This process was repeated for all additional pertinent articles identified from these articles, and so on, until no additional articles were identified. RESULTS: A total of 21 articles were identified. CONCLUSIONS: Occipital nerve block is an effective treatment for cervicogenic headache, cluster headache, and occipital neuralgia. While a double blinded randomized placebo controlled clinical trial is lacking, multiple open label studies reported favorable results for migraine. Two other possible uses of ONB worthy of further study are use as a rescue treatment and as an adjunctive treatment for medication overuse headache. ONB may be effective for tension headache, but only under very specific circumstances. ONB is either ineffective or only effective under as yet unstudied circumstances for hemicrania continua and chronic paroxysmal hemicrania. Some practitioners use occipital nerve (ON) tenderness to palpation (TTP) or reproduction of headache pain with ON pressure (RHPONP) as selection criteria for identifying appropriate patients. While only a clinical trial can produce a definitive answer, current evidence suggests that these selection criteria are not necessary for cervicogenic headache or cluster headache. Occipital neuralgia by definition involves TTP of the ONs. Whether RHPONP or ON TTP predicts success in migraine is unclear, and may relate to whether steroids are used. A single blinded randomized controlled trial evaluating local anesthetic with steroids vs local anesthetic alone for transformed migraine reported slightly worse results with steroids, but there are several alternate explanations for this finding other than steroids being counterproductive. The technique of repetitive ONBs deserves further study. PMID- 19674127 TI - Presence of high numbers of transcriptionally active Helicobacter pylori in vomitus from Bangladeshi patients suffering from acute gastroenteritis. AB - BACKGROUND: Helicobacter pylori is one of the most prevalent human bacterial pathogens; however, its transmission pathways remain unknown. New infections of H. pylori during outbreaks of gastroenteritis have been suggested previously, and to explore this transmission route further H. pylori was quantified in vomitus and diarrheal stool of patients suffering from acute gastroenteritis in Dhaka, Bangladesh. MATERIALS AND METHODS: Vomitus and stool samples from 28 patients seeking care at the International Centre for Diarrhoeal Disease Research hospital were analyzed for presence of H. pylori and other pathogens using quantitative culturing, real-time polymerase chain reaction (PCR), and H. pylori stool antigen test. Bacterial gene expression was analyzed using reverse transcriptase real time PCR. RESULTS: The results of real-time PCR show that 23 (88%) of the 26 vomitus samples and 17 (74%) of the 23 stool samples were H. pylori positive, while stool antigen test show that 14 (67%) of the 21 stool samples were H. pylori positive. H. pylori could not be isolated by culture. Analysis using quantitative culture and real-time PCR to detect Vibrio cholerae showed strong correlation between these methods, and validating real-time PCR. Analysis of H. pylori virulence gene transcription in vomitus, diarrheal stool, antral and duodenal biopsy specimens, and in vitro cultures showed that cagA, flaA, and ureA were highly transcribed in vomitus, biopsy specimens, and cultures, whereas hpaA and vacA were expressed at lower levels. No H. pylori gene expression was detected in diarrheal stool. CONCLUSIONS: We conclude that high numbers of transcriptionally active H. pylori are shed in vomitus, which indicates that new infections may be disseminated through vomiting. PMID- 19674128 TI - Distinctiveness of the cagA genotype in children and adults with peptic symptoms in South China. AB - BACKGROUND: Helicobacter pylori infection is different between children and adults, not only in infection rate but also in virulence genotypes. However, the 3' region of CagA, important in stomach carcinogenesis, still remains unclear in children. The present study aims to compare the frequency of cagA and the distribution of its subtypes between children and adults in South China. MATERIALS AND METHODS: One hundred and twenty-eight children and 99 adults with peptic symptoms were enrolled in our research. Histology, rapid urease test, and real-time polymerase chain reaction (PCR) assay were used to diagnose H. pylori infection. vacA s1 was detected by real-time PCR, and EPIYA motifs in the 3' region of CagA by conventional PCR and DNA sequencing. RESULTS: H. pylori infection was diagnosed in 53 children and 62 adults. vacA s1 was identified in 90.6% and 91.9% of infected children and adults, respectively. Furthermore, cagA was identified in 73.6% and 82.3% of infected children and adults, respectively. No patient with multiple cagA subtypes was observed. A higher prevalence of more virulent cagA genotype was found in children compared to adults (p < .05). Thirty eight of 39 (97.4%) cagA-positive children were found to have EPIYA-ABD and only one (2.6%) with EPIYA-ABC. In adults, four types of EPIYA motifs--ABC (29.4%), ABD (64.7%), ABAB (2%), and AAD (3.9%)--were identified, and the ABD type was found more commonly in severe diseases, such as atrophic gastritis (53.3%) and gastric cancer (71.4%). CONCLUSION: cagA genotypes in children and in adults are different, and EPIYA-ABD may have potential clinical implication in the development of gastric cancer in South China. PMID- 19674129 TI - Distinct cagA EPIYA motifs are associated with ethnic diversity in Malaysia and Singapore. AB - BACKGROUND: In vitro studies have shown that the biologic activity of CagA is influenced by the number and class of EPIYA motifs present in its variable region as these motifs correspond to the CagA phosphorylation sites. It has been hypothesized that strains possessing specific combinations of these motifs may be responsible for gastric cancer development. This study investigated the prevalence of cagA and the EPIYA motifs with regard to number, class, and patterns in strains from the three major ethnic groups within the Malaysian and Singaporean populations in relation to disease development. MATERIALS AND METHODS: Helicobacter pylori isolates from 49 Chinese, 43 Indian, and 14 Malay patients with functional dyspepsia (FD) and 21 gastric cancer (GC) cases were analyzed using polymerase chain reaction for the presence of cagA and the number, type, and pattern of EPIYA motifs. Additionally, the EPIYA motifs of 47 isolates were sequenced. RESULTS: All 126 isolates possessed cagA, with the majority encoding EPIYA-A (97.6%) and all encoding EPIYA-B. However, while the cagA of 93.0% of Indian FD isolates encoded EPIYA-C as the third motif, 91.8% of Chinese FD isolates and 81.7% of Chinese GC isolates encoded EPIYA-D (p < .001). Of Malay FD isolates, 61.5% and 38.5% possessed EPIYA-C and EPIYA-D, respectively. The majority of isolates possessed three EPIYA motifs; however, Indian isolates were significantly more likely to have four or more (p < .05). CONCLUSION: Although, H. pylori strains with distinct cagA-types are circulating within the primary ethnic groups resident in Malaysia and Singapore, these genotypes appear unassociated with the development of GC in the ethnic Chinese population. The phenomenon of distinct strains circulating within different ethnic groups, in combination with host and certain environmental factors, may help to explain the rates of GC development in Malaysia. PMID- 19674130 TI - Activities of urease and nickel uptake of Helicobacter pylori proteins are media- and host-dependent. AB - BACKGROUND: Nickel-dependent urease activity and nickel supply are essential for successful colonization of Helicobacter pylori in the acidic environment of the human stomach. A comparison of media effects on these two activities have never been carried out. Additionally to H. pylori we cultivated an Escherichia coli strain expressing the urease and the nickel transporter NixA of H. pylori on the same four media and measured in all cases urease and nickel uptake activity. AIM: To compare nickel uptake and urease activity on an inter- and intraspecies level. RESULTS: In H. pylori nickel uptake (four to 200 times) and urease activities (400 to 30,000 times) were found to be much higher in comparison to the tested E. coli strain after growth on all media. These differences could not be explained by reduced protein amounts in the heterologous host E. coli. On which media the two bacteria extracted most of the nickel were organism-dependent: E. coli on Brucella Broth, H. pylori on Trypticase Soy Broth, and Minimal Media. CONCLUSION: H. pylori took nickel much more efficiently up than E. coli. The observed differences in urease activity are most likely due to additional protein components absent in the recombinant E. coli strain. The observed variety in nickel uptake and urease activities on the different media in the same organism depended on the intrinsic nickel content and chelating capacities of media components. Different culture conditions may lead to varying results; generalizations should be concluded only after excluding their media dependence. PMID- 19674131 TI - Helicobacter pylori VacA activates NF-kappaB in T cells via the classical but not alternative pathway. AB - BACKGROUND: Helicobacter pylori secretes vacuolating cytotoxin (VacA) that damages the gastric epithelium by erosion and loosening of tight junctions. VacA has also immunosuppressive effects, inhibiting interleukin (IL)-2 secretion by interference with the T cell receptor/IL-2 signaling pathway. This study investigated the effect of VacA on gene expression of T cells. MATERIALS AND METHODS: Gene expression profile of a T cell line, Jurkat, was analyzed by the cDNA microarray technique after VacA challenge. The expression of specific mRNAs was assessed by reverse transcription-polymerase chain reaction. Interleukin (IL) 8 concentrations in culture supernatants and cell surface expression of CD69 were measured by enzyme-linked immunosorbent assay and flow cytometry, respectively. We evaluated nuclear factor-kappaB (NF-kappaB) activation in Jurkat cells challenged with VacA by luciferase assay, electrophoretic mobility shift assay, and Western blot analysis. RESULTS: VacA produced two or greater fold up regulation of expression of 60 genes. Most of these genes were associated with signal transduction, regulation of gene expression, apoptosis, and inflammation. Up-regulation of four genes (IL8, IL2RA, ICAM1, and CD69) was confirmed. The supernatants of cells incubated with VacA showed significantly higher secretion levels of IL-8 than those incubated without VacA. VacA also induced the cell surface expression of CD69. Since microarray analysis indicated NF-kappaB was involved in the transcriptional activation of the above genes, we examined NF kappaB signaling pathway. VacA activated NF-kappaB via classical but not alternative pathway. CONCLUSIONS: VacA has two paradoxical effects on T cells, immunosuppression, and proinflammatory effects. The latter is mediated by NF kappaB activation. PMID- 19674133 TI - Epidemiology of Helicobacter pylori infection in asymptomatic children: a prospective population-based study from the Czech Republic. Application of a monoclonal-based antigen-in-stool enzyme immunoassay. AB - BACKGROUND: Acquisition of Helicobacter pylori occurs mainly in childhood and is significantly influenced by geographical variations. The aim of this study is to evaluate the prevalence of H. pylori infection in a population-based sample of asymptomatic children in the Czech Republic. Furthermore, this study aims to identify potential risk factors associated with this infection. MATERIALS AND METHODS: A prospective, cross-sectional, population-based study was undertaken in 1545 asymptomatic Czech children (aged 0-15 years; male 49.3%). Active H. pylori infection was diagnosed by monoclonal antibody-based antigen-in-stool enzyme immunoassay. Socio-demographic details of each subject were analyzed using a self administered standardized questionnaire. Multiple regression analysis was performed. RESULTS: Overall, 7.1% of asymptomatic children were diagnosed with H. pylori infection. Of the infected children, 5.8% lived in the general population. A positive association was found with increasing age, although not with gender. Independent risk factors associated with H. pylori infection in our pediatric population were: the number of children in a household (odds ratio [OR] 4.26; confidence interval [CI] 1.91-9.80); lack of formal education of fathers (OR 0.23; CI 0.18-0.64) and institutionalized children (OR 6.33; CI 2.25-26.50). CONCLUSIONS: This study of a large cohort of children demonstrated that, independent of gender, H. pylori infection in the Czech Republic is among the lowest reported in Europe. Socioeconomically disadvantaged children, unfortunately, are still at risk of harboring this potentially preventable infection in this low-prevalence region. PMID- 19674132 TI - Oxidative stress in Helicobacter pylori infection: does supplementation with vitamins C and E increase the eradication rate? AB - AIM: This study aims to assess the antioxidant property of vitamins E and C in Helicobacter pylori infection, and to determine if adding them to standard triple therapy plus bismuth subcitrate increases the H. pylori eradication rate. METHODS: This study included 160 patients infected with H. pylori, who were randomized into one of two groups. Patients in group A (n = 80) received lansoprazole (30 mg, b.i.d.), amoxicillin (1000 mg, b.i.d.), clarithromycin (500 mg, b.i.d.), and bismuth subcitrate (300 mg, q.i.d.) for 14 days, while patients in group B (n = 80) received vitamin C (500 mg, b.i.d.) and vitamin E (200 IU, b.i.d.) for 30 days, in addition to lansoprazole (30 mg, b.i.d.), amoxicillin (1000 mg, b.i.d.), clarithromycin (500 mg, b.i.d.), and bismuth subcitrate (300 mg, q.i.d.) for 14 days. Total antioxidant capacity (TAC) was evaluated with a Randox kit. Success rate was calculated using both intention-to-treat (ITT) and per-protocol (PP) analyses. RESULTS: One hundred and sixty patients were analyzed using ITT analysis. One hundred and fifty-three patients completed the study. In group A, H. pylori eradication was achieved in 48 (60%) of the 80 patients included in the ITT analysis, and in 48 (64%) of the 75 patients included in the PP analysis. In group B, H. pylori eradication was achieved in 73 (91.25%) of the 80 included in the ITT analysis and in 73 (93.5%) of the 78 patients included in the PP analysis. The eradication rate was significantly higher in group B than in group A (p < .005). TAC was at the lower limit of normal in both groups and the difference between them was not statistically significant (p > .05). CONCLUSION: In group B, H. pylori eradication rate was 91.25%, which is higher than the ideal 80% eradication rate. The results of the present study show that adding the prescribed doses of vitamins E and C to antimicrobial therapy is effective in eradicating H. pylori infection. PMID- 19674134 TI - Helicobacter pylori seroprevalence in symptomatic veterans: a study of 7310 patients over 11 years. AB - BACKGROUND AND AIMS: The prevalence of Helicobacter pylori infection has been decreasing in the USA, but recent data are lacking. This study evaluates the seroprevalence for anti-H. pylori antibodies in symptomatic veterans tested over the past 11 years. MATERIALS AND METHODS: The same serum anti-H. pylori IgG detection system has been used at a tertiary care Veterans Affairs hospital since late 1996. Results of all tests performed from 1997 to 2007 were analyzed. RESULTS: Of 7310 unique patients tested, 3982 (54.5%) were positive. Seropositivity declined from 70.8% in 1997 to 48.6% in 2002, then reached a plateau around 50%. A strong birth cohort effect was present, from a seropositivity of 72.7% for the veterans born before 1920 to 22% for those born between after 1980. CONCLUSIONS: Despite a constant birth cohort effect, H. pylori seropositivity among symptomatic veterans leveled down at approximately 50% after declining steadily from 1997 to 2002. PMID- 19674135 TI - Human gastrin: a true growth factor for Helicobacter pylori? PMID- 19674136 TI - Chronic gastritis develops and progresses over the years under permanent exposure of Helicobacter pylori: is the case different in Mozambican dyspeptic patients? PMID- 19674157 TI - Receptor for activated C-kinase 1 regulates the cellular localization and function of ABCB4. AB - AIM: Multidrug resistance protein 3 (MDR3/ABCB4), located on the bile canalicular membrane of hepatocytes, is responsible for the translocation of phosphatidylcholine across the plasma membrane, and its hereditary defect causes liver disorders, such as progressive familial intrahepatic cholestasis type 3. We aimed to identify the proteins responsible for the surface expression of human ABCB4. METHODS: We performed yeast two-hybrid screening with the cytoplasmic linker region of ABCB4 against a human liver cDNA library. This screening allowed us to identify the receptor for activated C-kinase 1 (RACK1) as a novel binding partner of ABCB4. The association of RACK1 with the linker region of ABCB4 was further confirmed by GST-pulldown assay, although we could not find out the interaction of full length of ABCB4 and RACK1 in co-immunoprecipitation assay in HeLa cells. RESULTS: Down-regulation of endogenous RACK1 expression by siRNA in HeLa cells resulted in the localization of ABCB4 in the cytosolic compartment as well as reduced protein expression of ABCB4, although mRNA expression and the protein stability of ABCB4 were not affected by the suppression of endogenous RACK1. Similar alterations in cellular localization of ABCB4 were also found by suppressing endogenous RACK1 expression in HepG2 cells. Consequently, ABCB4 mediated phosphatidylcholine translocation activity was significantly reduced when endogenous RACK1 expression was suppressed in HeLa cells. In contrast, the membrane surface localization and the protein expression of ABCB1 were not affected by the suppression of endogenous RACK1 expression. CONCLUSION: These results suggest that RACK1 may have a functional significance as a regulatory cofactor of ABCB4 and is indispensable for the plasma membrane localization and translocation function of ABCB4. PMID- 19674158 TI - Zoonotic risk of hepatitis E virus (HEV): A study of HEV infection in animals and humans in suburbs of Beijing. AB - AIM: To investigate hepatitis E virus (HEV) infection among different animals and workers in pig farms and slaughterhouses, and analyze the genotype of HEV isolated in this study. METHODS: Serum samples were collected from adult swine, cows, sheep, younger swine (< 3 months), and workers in pig farms and slaughterhouses (professional group). Fecal samples were collected from younger swine in the south suburbs of Beijing. Anti-HEV antibody was evaluated by direct sandwich enzyme immunoassay. HEV RNA was extracted from fecal samples and amplified by nested reverse transcription polymerase chain reaction (RT-nPCR). The PCR products were sequenced, and the sequence homology and phylogenetics of the HEV strains isolated from swine were analyzed. RESULTS: The anti-HEV positivity rates in adult swine, cows, sheep, younger swine, professional group and general population were 98.23% (222/226), 29.35% (54/184), 9.80% (20/207), 60.73% (99/164), 42.51% (105/247) and 20.29% (522/2572), respectively. The HEV RNA positivity rate of fecal samples was 22.89% (19/83) and 16/19 samples were positive for HEV RNA amplified with both primers, HEV open reading frame (ORF)1 and HEV ORF2. Sequence analysis of these 16 samples showed that there were two groups, designated BJ-1 and BJ-2. The nucleotide homology of BJ-1 and BJ-2 was 99%. Phylogenetic analysis indicated that both of these groups belonged to genotype 4d. CONCLUSION: Workers in pig farms and slaughterhouses were more likely to contract HEV infection than the general population because of close contact with swine with a high prevalence of anti-HEV. PMID- 19674159 TI - High expression of angiotensin-converting enzyme and angiotensin-converting enzyme 2 in preservation injury after liver transplantation in rats. AB - AIM: To explore the possible role of local renin-angiotensin system (RAS) in preservation injury (PI) after liver transplantation by studying expression of angiotensin-converting enzyme (ACE) and angiotensin-converting enzyme 2 (ACE2) in the transplanted liver of rats and its relationship with PI. METHODS: The animals receiving liver transplantation were assigned to cold preservation group (CP group) and non-cold preservation group (NCP group). The sham-operation group was used as the control. The severity of PI was assessed by histology. The mRNA and protein expressions of ACE and ACE2 were detected by real-time PCR, Western blot, respectively. Tissue hypoxia was assessed by pimonidazole staining. RESULTS: Various degrees of tissue injury were observed after liver transplantation, especially in CP group. ACE2 mRNA and protein expressions in the transplantation groups were elevated significantly compared with those of the control group (P < 0.01, P < 0.05), and higher in CP group than those in NCP group (P < 0.05). There was a close positive correlation between PI and mRNA expression of ACE and ACE2. Positive pimonidazole staining distributed around the hepatic central vein, and became darker and more extensive with deterioration of PI. CONCLUSION: ACE2 was closely related to tissue hypoxia due to CP-induced PI of the transplanted liver, and ACE may aggravate the inflammation in PI. Local RAS may play an important role in PI of the transplanted liver. PMID- 19674161 TI - Letter to the Editor on 'Effects of exposure to a mobile phone on testicular function and structure in adult rabbit' by Salama et al. PMID- 19674162 TI - The personalisation of the British National Health Service: empowering patients or exacerbating inequality? PMID- 19674163 TI - Analysis of high-resolution HLA-A, -B, -Cw, -DRB1, and -DQB1 alleles and haplotypes in 718 Chinese marrow donors based on donor-recipient confirmatory typings. AB - High-resolution human leucocyte antigen (HLA)-A, -B, -Cw, -DRB1, and -DQB1 alleles and haplotype frequencies were analysed from 718 Chinese healthy donors selected from the Chinese Marrow Donor Program registry based on HLA donor recipient confirmatory typings. A total of 28 HLA-A, 61 HLA-B, 30 HLA-Cw, 40 HLA DRB1 and 18 HLA-DQB1 alleles were identified, and HLA-A*1101, A*2402, A*0201, B*4001, Cw*0702, Cw*0102, Cw*0304, DRB1*0901, DRB1*1501, DQB1*0301, DQB1*0303 and DQB1*0601 were found with frequencies higher than 10% in this study population. Multiple-locus haplotype analysis by the maximum-likelihood method revealed 45 A B, 38 Cw-B, 47 B-DRB1, 29 DRB1-DQB1, 24 A-B-DRB1, 38 A-Cw-B, 23 A-Cw-B-DRB1, 33 Cw-B-DRB1-DQB1 and 22 A-Cw-B-DRB1-DQB1 haplotypes with frequencies >0.5%. The most common two-, three-, four- and five-locus haplotypes in this population were: A*0207-B*4601 (7.34%), Cw*0102-B*4601 (8.71%), B*1302-DRB1*0701 (6.19%), DRB1*0901-DQB1*0303 (14.27%), A*3001-B*1302-DRB1*0701 (5.36%), A*0207-Cw*0102 B*4601 (7.06%), A*3001-Cw*0602-B*1302-DRB1*0701 (5.36%), Cw*0602-B*1302-DRB1*0701 DQB1*0202 (6.12%) and A*3001-Cw*0602-B*1302-DRB1*0701-DQB1*0202 (5.29%). Presentation of the high-resolution alleles and haplotypes data at HLA-A, -B, Cw, -DRB1 and -DQB1 loci will be useful for HLA matching in transplantation as well as for other medical and anthropological applications in the Chinese population. PMID- 19674164 TI - Near infrared spectroscopy study of the central nervous activity during artificial changes in bladder sensation in men. AB - OBJECTIVES: To investigate the regional specificity of multi-channel near infrared spectroscopy (NIRS) on the detection of urination-related cortical activation in healthy men. METHODS: Frontal lobe blood flow was measured non invasively in 20 resting subjects using NIRS before and after micturition. The relative concentrations of oxyhemoglobin were recorded over the frontal lobe during artificial changes in bladder sensation with a 52-channel NIRS machine. An increase in oxyhemoglobin represented locally increased blood flow. In artificially induced bladder sensation (desire to void), the lower abdomen of each participant was pressed by the examiner's hand to keep each participant feeling the desire to void for 5 s. Each participant received this series of tasks three times before and after urination. Activated concentrations calculated by contrast of subtracting the two different conditions were designed to disclose the brain areas that are involved during artificial changes in bladder sensation. The measurements were repeatedly carried out for each subject on different days to reveal the reproducibility and intra-individual variability of the results. RESULTS: Cerebral responses during artificial changes in bladder sensation were bilaterally associated with increased levels of oxyhemoglobin in the frontal area. Oxyhemoglobin increases during the compression maneuver were significantly larger than those during an empty bladder in several bilateral frontal channels (P < 0.05). CONCLUSIONS: Frontal regions were activated during artificial changes in bladder sensation before micturition using a 52-channel NIRS machine. PMID- 19674165 TI - Efficacy and safety of dutasteride in Japanese men with benign prostatic hyperplasia. AB - OBJECTIVES: To assess the efficacy and safety of dutasteride in Japanese men with benign prostatic hyperplasia (BPH). METHODS: This was a randomized, double-blind, placebo-controlled, parallel-group study. A total of 378 subjects with clinical BPH having an International Prostate Symptom Score (IPSS) of 8 points or greater, a prostate volume of 30 mL or greater, and a maximal urinary flow rate (Qmax) of 15 mL/s or less were randomized to receive placebo or dutasteride once daily for 52 weeks. Subjects were stratified according to tamsulosin use at baseline. The numbers of subjects with and without tamsulosin use were 242 and 136, respectively. IPSS, Qmax, prostate volume and drug safety were evaluated. RESULTS: Continued improvement in IPSS was noted in the dutasteride group, and dutasteride significantly decreased IPSS compared with placebo. At week 52, dutasteride significantly improved Qmax and prostate volume compared with placebo. Drug-related sexual function events in the dutasteride group were infrequent and generally were not treatment limiting. CONCLUSIONS: Dutasteride improves urinary symptoms and flow rate and reduces prostate volume. In Japanese men with BPH, it is effective and generally well tolerated during the one-year treatment period. PMID- 19674166 TI - Tunica albuginea urethroplasty for anterior urethral strictures: a urethroscopic analysis. AB - OBJECTIVE: To assess the efficacy of tunica albuginea urethroplasty (TAU) for anterior urethral strictures. METHODS: We assessed 206 patients with anterior urethral strictures who underwent TAU. The procedure involves mobilization of strictured urethra and laying it open with a dorsal slit. Edges of the slit-open urethra are sutured to edges of the urethral groove with a silicon catheter in situ. Thus in neourethra, the roof is formed by tunica albuginea of the urethral groove. Results were assessed at 6, 12, 24 and 36 months by comparative analysis of patient satisfaction along with retrograde urethrogram, urethrosonogram, uroflowmetry, and were categorized as good, fair and poor. Good and fair results were considered as successful. Thirty patients were taken for postoperative urethroscopic analysis to allow better understanding of both successful and failed cases. RESULTS: Postoperative evaluation at 6 months showed a 96.6% success rate, which decreased to 94.7% at 1 year, 93.2% at 2 years and over 90% at the end of 3 years. The overall failure rate was 9.2%, which required revision surgery. Urethroscopic visualization of the reconstruction site showed wide, patent and distensible neourethra uniformly lined by urothelium over roof formed by tunica albuginea of the corpora cavernosa in successful cases. Failure cases showed diffuse fibrotic narrowing or circumferential scarring. CONCLUSION: Tunica albuginea is a locally available distensible tissue, sufficient to maintain the patency of the neourethra, without any graft or flap. TAU is easier and useful when patients have unhealthy oral mucosa due to tobacco chewing. PMID- 19674168 TI - Prognostic factors for cases with metastatic renal cell carcinoma in the era of targeted medicine. AB - Prognostic factors in the setting of treated metastatic renal cell carcinoma (mRCC) can theoretically predict drug response, progression-free survival or overall survival. These factors are potentially useful for informing patients and for tailoring medical treatment according to risk assessment. Prognostic factors were well defined in the era of immunotherapy. Since 2006, tyrosine kinase inhibitors and anti-angiogenic drugs have revolutionized the treatment of mRCC by improving progression-free survival and overall survival. Physicians now need new predictive tools adapted to targeted therapies. The aim of our study was to review the current knowledge about prognostic factors in mRCC by focusing on anatomical, clinical, biological, histological, radiological and molecular parameters. The most recent integrated prognostic models will be reviewed as well. PMID- 19674167 TI - Radical prostatectomy for high-risk prostate cancer: biochemical outcome. AB - OBJECTIVES: To determine the biochemical outcome following radical prostatectomy alone in patients with high-risk prostate cancer. METHODS: Between January 2002 and August 2007, 252 patients underwent radical retropubic prostatectomy. Those who received neoadjuvant hormone therapy were excluded from this analysis. Based on pre-operative data, we stratified the patients into low, intermediate, and high-risk groups according to the risk criteria of the National Comprehensive Cancer Network in 2003, respectively. Prostate-specific antigen (PSA) failure was defined as any detectable PSA level higher than 0.2 ng/mL. RESULTS: The PSA failure-free survival rate for the high-risk group (n = 46) was 64.5% after a median follow-up period of 39 months. Among patients with high-risk disease, none with pathologically organ-confined cancer (n = 19) and a negative surgical margin had PSA failure. The PSA failure-free rate in patients with non organ-confined cancer (n = 27) was 39.5%. Among the pretreatment variables, a positive biopsy core percentage (the number of positive biopsy cores/total biopsy core) >or=30 was a significant independent predictor of extra prostatic extension. CONCLUSIONS: Radical prostatectomy is feasible in high-risk prostate cancer patients, only if they have a pathologically organ-confined disease. PMID- 19674169 TI - Current status of ureteroscopic treatment for urolithiasis. AB - Intracorporeal treatment of urolithiasis is characterized by continuous technological evolution. In this review we present updated data upon the use of ureteroscopy for the management of urolithiasis. Novel digital flexible ureteroscopes are used in clinical practice. Ureteroscopic working tools are revolutionized resulting in safer and more efficient procedures. Special categories of stone patients such as pregnant women, children and patients on anticoagulation medication can now undergo uneventful ureteroscopy. Routine insertion of stents and access sheaths as well as bilateral ureteroscopy is still a controversial issue. Future perspectives include smaller and better instruments to visualize and treat a stone, while robotic ureteroscopy is becoming a fascinating reality. PMID- 19674170 TI - Factors affecting quality of life in Taiwanese survivors of childhood cancer. AB - AIM: This paper is a report of a pilot study to examine the significance, direction, and magnitude of the relationship of risk factors, protective factors, and resilience to quality of life in survivors of childhood cancer, and to explore what these variables meant to participants in order to better understand the quantitative findings and to assess the validity of the instruments with this population. BACKGROUND: Cancer survivors contend with physical, psychosocial and emotional consequences of the disease and its treatment. There has been little research in Taiwan assessing these outcomes and, as survivor numbers increase, appropriate interventions need to be implemented. Haase's Adolescent Resilience Model was used as the theoretical framework. METHODS: A mixed methods design was used to assess the relationship between the variables for survivors of leukaemia and brain cancers. Ninety-eight participants completed quantitative tools and interviews, and the data were collected in 2006. FINDINGS: Survivors of leukaemia had better quality of life than those with brain tumours. This is attributed to more positive protective factors, fewer illness-related and individual risk factors and more positive resilience scores. Findings were congruent with similar studies from other countries. Qualitatively, participants believed that better quality of life meant less loss of self, a more integrated self, less chronic fear, and an ability to rebound and achieve personal control over one's life. CONCLUSION: Interventions and assessment of their efficacy are necessary to help survivors capitalize on their strengths, to decrease their individual risks and to promote resilience promote self-control. PMID- 19674171 TI - The role of irrational thought in medicine adherence: people with diabetic kidney disease. AB - AIM: This paper is a report of a study conducted to examine how irrational thinking affects people's adherence to multiple medicines prescribed to manage their diabetic kidney disease. BACKGROUND: Approximately 50% of people are non adherent to their prescribed medicines and the risk of non-adherence escalates as the number of prescribed medicines increases. Adherence to prescribed medicines can slow disease progression in diabetic kidney disease. METHODS: A descriptive exploratory design was used. In-depth interviews were conducted with 23 participants recruited from a nephrology outpatient clinic in Australia in 2007. Data were analysed using a 'framework' method. FINDINGS: Participants' mean age was 59 years, they had approximately six chronic conditions in addition to their diabetic kidney disease and were prescribed a median of ten medicines daily. Two major themes of irrational thinking--heuristics and denial--and subthemes were identified. Heuristics contributed to inaccurate risk assessment and biases affecting rational judgement concerning medicines, whereas denial was used to enhance coping necessary to manage this complex health condition. CONCLUSION: Participants underestimated their health risks because they had been taking medicines for many years and preferred not to dwell on their ill health. A large amount of irrational thinking was related to maintaining the emotional strength necessary to manage their comorbid conditions as best they could. Regular assessment and support of medicine adherence throughout the disease course is necessary to avert the development of counterproductive heuristics and denial affecting medicine adherence. PMID- 19674172 TI - Work sampling: a quantitative analysis of nursing activity in a neuro rehabilitation setting. AB - AIM: The aim of this investigation was to establish the distribution and proportion of nursing activity represented by patient-related care activities (direct and indirect), and other nursing activities (unit-related and personal) within one inpatient neurological rehabilitation unit. BACKGROUND: A set of tools has been developed for estimating the care/nursing hours required for direct hands-on patient care in hospital rehabilitation settings. However, to apply this information to estimate the actual staffing requirements in relation to a given caseload, it is necessary to know the proportion of nursing workload assigned to other activities and how this may vary throughout the day. METHOD: A work sampling study was conducted during 2004. A snapshot of nursing activity was recorded at 5-minute intervals from 0600 to 2355 spread over 2 weeks, with one session from 0600 to 1525 and the second from 1530 to 2355. RESULTS: A total of 8883 nursing activities were observed and recorded over 126 hours and categorized as follows: 4060 (46%) direct patient care, 2218 (25%) indirect patient care, 874 (10%) unit-related and 1731 (19%) personal time. The proportions of direct care fluctuated throughout the day, with direct care activities mainly concentrated in early mornings and to a lesser extent evenings. CONCLUSION: Direct patient care accounted for less than half of the nursing activity in a rehabilitation setting. Estimates of staffing requirement must also take account of the time required for indirect care and non-patient related activity. PMID- 19674173 TI - Emotional stability of nurses: impact on patient safety. AB - AIM: This paper is a report of a study conducted to examine the influence of the emotional stability of nurses on patient safety. BACKGROUND: Individuals with greater emotional stability are less likely to exhibit strong emotional reactions to stressful situations, and tend to be more proactive and successful in problem solving. Effectively managing patient safety is a priority concern in countries where nurses face high pressure. A heavy work load leads to burnout (a syndrome associated with negative emotions), reduced job satisfaction and increased turnover. While emotional stability influences job performance in various contexts, its influence on patient safety has not been addressed. METHOD: A cross sectional design was adopted. The sample comprised 263 nurses working in two Taiwanese medical centres. The data were collected in 2007-2008, with a response rate of 926%. All participants were nursing college graduates aged below 50 years. Participants provided information on both their emotional stability and patient safety. Staffing adequacy, hospital, and years of nursing experience served as control variables. FINDINGS: Hierarchical regression analysis indicated that emotional stability predicted patient safety (beta = 0.18, P < 001). The addition of emotional stability as a predictor of patient safety increased the associated explained variance (deltaR2 = 0.03, P < 0.01). CONCLUSION: It is important for to managers create an organisational climate that promotes the emotional stability of nurses. This could help to improve global patient safety by reducing the frequency of adverse events. PMID- 19674174 TI - Administrative claims data analysis of nurse practitioner prescribing for older adults. AB - AIM: This paper is a report of a study to identify the patterns of prescribing by primary health care nurse practitioners for a cohort of older adults. BACKGROUND: The older adult population is known to receive complex pharmacotherapy. Monitoring prescribing to older adults can inform quality improvement initiatives. In comparison to other countries, research examining nurse practitioner prescribing in Canada is limited. Nurse practitioner prescribing for older adults is relatively unexplored in the international literature. Although commonly used to study physician prescribing, few studies have used claims data from drug insurance programmes to investigate nurse practitioner prescribing. METHOD: Drug claims for prescriptions written by nurse practitioners from fiscal years 2004/05 to 2006/07 for beneficiaries of the Nova Scotia Seniors' Pharmacare programme were analysed. Data were retrieved and analysed in May 2008. Prescribing was described for each drug using the World Health Organization Anatomical Therapeutic Chemical code classification system by usage and costs for each fiscal year. RESULTS: Antimicrobials and non-steroidal anti-inflammatory drugs consistently represented the top ranked groups for prescription volume and cost. Over the three fiscal years, antimicrobial prescription rates declined relative to rates of other groups of medications. Prescription volume per nurse doubled and cost per prescription increased by approximately 20%. CONCLUSION: Prescription claims data can be used to characterize the prescribing trends of nurse practitioners. Research linking patient characteristics, including diagnoses, to prescriptions is needed to assess prescribing quality. Some potential areas of improvement were identified with antimicrobial and non steroidal antiinflammatory selection. PMID- 19674175 TI - Relationship of working mothers' parenting style and consistency to early childhood development: a longitudinal investigation. AB - AIM: This paper is a report of a longitudinal study of the relationship of working mothers' parenting style to their children's social competence and vocabulary/ motor/intellectual development. BACKGROUND: With an increasing number of women choosing to remain in the workforce after starting a family, there has been a concomitant increase in use of non-parental childcare facilities to help look after the child while the mother is at work. This increase in non-parental care has led to a dramatic change in the traditional child-rearing environment. METHODS: Long-term investigations were conducted over a period of 2 years in 41 Japanese government-licensed childcare facilities. Child development was evaluated by childcare professionals and parenting style was assessed by questionnaire. A total of 504 children and their mothers participated in the study. Data collection was carried out in 2004 and 2006. FINDINGS: We found that the changes in parenting style were statistically significantly related to children's development after 2 years. For instance, changes in the parent-child playing routine contributed to the child's social competence (odds ratio = 11.088). Variation in working mothers' disciplinary practices was also associated with children's vocabulary development after 2 years (odds ratio = 2246). CONCLUSION: Working mothers should increase interactions with their children in their free time to reduce the risk of developmental delay. Daily childcare support provided by family members or social organizations for long-term working mothers is helpful in mediating the negative relationship of mothers' working with children's development. PMID- 19674176 TI - Instruments to tailor care of people with type 2 diabetes. AB - AIM: This paper is a report of a study conducted to assess the effectiveness of an educational intervention for people with type 2 diabetes based on self completed instruments to identify particular areas of self-care needs. BACKGROUND: Diabetes is a demanding disease which requires self-care. Instruments that identify self-care needs or factors affecting self-care can be helpful to detect level of self-care, distress or knowledge among individuals with diabetes. METHODS: Participants were randomized into intervention (n = 28) or control groups (n = 25). Both groups answered five validated instruments three times, at baseline and after 3 and 6 months, and biological measurements were conducted simultaneously. The intervention was based on an empowerment approach. The study started in November 2005 and lasted until March 2007. FINDINGS: There was no statistically significant difference between groups in level of glycated haemoglobin postintervention as it reduced statistically significantly in both groups between baseline and 3 months but increased again after 6 months. There was no statistically significant difference between groups in body mass index reduction; the intervention group achieved a statistically significant body mass index reduction but this was unchanged in the control group. No statistically significant differences between groups were found in scores for empowerment, well being and distress. There was a statistically significant difference between groups in knowledge postintervention. CONCLUSION: Use of instruments to identify self-care needs can enable patient-tailored care as it allows direct focusing on issues that are challenging and of relevance for each individual. It seems to be feasible to use the telephone to conduct an empowering educational intervention after one meeting. PMID- 19674177 TI - Diet selection in a molluscivore shorebird across Western Europe: does it show short- or long-term intake rate-maximization? AB - 1. Studies of diet choice usually assume maximization of energy intake. The well known 'contingency model' (CM) additionally assumes that foraging animals only spend time searching or handling prey. Despite considerable empirical support, there are many foraging contexts in which the CM fails, but such cases were considered exceptions rather than the rule. 2. For animals constrained by the rate at which food is digested, CM does not necessarily lead to maximal energy intake rates because the time for digestion is not part of the selection criteria. In the main model developed to explain diet choice under a digestive constraint, the 'digestive rate model' (DRM), time lost to digestive breaks is minimized so that energy intake over total time (searching, handling, digestive breaks) is maximized. 3. It is increasingly acknowledged that most animals may face digestive constraints as prey capture rates vary over time and as it would be a waste to carry around heavy digestive machinery that can rapidly process food under all circumstances: this is only needed in times of high demand, provided that enough food can be found. 4. In molluscivore shorebirds ingesting hard-shelled prey such as red knots (Calidris canutus), the predictions of DRM were held up so far, whereas those of CM were rejected. However, most tests were carried out under controlled experimental conditions. Red knots overwinter in coastal areas over much of Western Europe and we capitalized on this variation by comparing, during a single winter, observed diet composition with predictions of DRM, CM and a null model assuming no prey selection ('no-selection model', NSM). 5. The observed diets were best predicted by DRM followed by CM. NSM poorly predicted observed diet choice. Under the present conditions, diet choice based on DRM would on average have yielded an energy intake rate twice as large as one based on CM. By adjusting the size of their gizzard (held constant in the present simulations), red knots could have lifted their energy intake rate further. We suggest that application of the DRM can help many diet studies forward, especially those previously seen as exceptions to the classical CM-based rule. PMID- 19674178 TI - Synchronous provisioning increases brood survival in cooperatively breeding pied babblers. AB - 1. Behavioural synchrony typically involves trade-offs. In the context of foraging, for example, synchrony may be suboptimal when individuals have different energy requirements but yield net benefits in terms of increased foraging success or decreased predation risk. 2. Behavioural synchrony may also be advantageous when individuals collaborate to achieve a common goal, such as raising young. For example, in several bird species, provisioners synchronize nest-feeding visits. However, despite the apparent prevalence of provisioning synchrony, it is not known whether it is adaptive or what its function might be. 3. Here, we propose a novel explanation for provisioning synchrony: it increases brood survival by decreasing the number of temporally separate nest visits and accordingly the chance that the nest will be detected by predators. Using cooperatively breeding pied babblers, we showed experimentally that provisioners synchronized nest visits by waiting for another provisioner before returning to the nest. Brood survival increased with provisioning synchrony. Provisioners were more likely to synchronize feeding visits for older nestlings as they were louder and possibly more conspicuous to predators. Finally, provisioners in large groups were more likely to wait for other provisioners and synchronized a higher proportion of all visits than those in smaller groups. Thus, provisioning synchrony may be one mechanism by which large groups increase brood survival in this species. 4. This study highlights a novel strategy that birds use to increase the survival of young and demonstrates the advantages of coordinated behaviour in social species. PMID- 19674179 TI - Stochastic demography and population dynamics in the red kangaroo Macropus rufus. AB - 1. Many organisms inhabit strongly fluctuating environments but their demography and population dynamics are often analysed using deterministic models and elasticity analysis, where elasticity is defined as the proportional change in population growth rate caused by a proportional change in a vital rate. Deterministic analyses may not necessarily be informative because large variation in a vital rate with a small deterministic elasticity may affect the population growth rate more than a small change in a less variable vital rate having high deterministic elasticity. 2. We analyse a stochastic environment model of the red kangaroo (Macropus rufus), a species inhabiting an environment characterized by unpredictable and highly variable rainfall, and calculate the elasticity of the stochastic growth rate with respect to the mean and variability in vital rates. 3. Juvenile survival is the most variable vital rate but a proportional change in the mean adult survival rate has a much stronger effect on the stochastic growth rate. 4. Even if changes in average rainfall have a larger impact on population growth rate, increased variability in rainfall may still be important also in long-lived species. The elasticity with respect to the standard deviation of rainfall is comparable to the mean elasticities of all vital rates but the survival in age class 3 because increased variation in rainfall affects both the mean and variability of vital rates. 5. Red kangaroos are harvested and, under the current rainfall pattern, an annual harvest fraction of c. 20% would yield a stochastic growth rate about unity. However, if average rainfall drops by more than c. 10%, any level of harvesting may be unsustainable, emphasizing the need for integrating climate change predictions in population management and increase our understanding of how environmental stochasticity translates into population growth rate. PMID- 19674180 TI - Sex-specific fitness correlates of dispersal in a house sparrow metapopulation. AB - 1. Dispersal affects many important ecological and evolutionary processes. Still, little is known about the fitness of dispersing individuals. 2. Here, we use data from a long-term study of a house sparrow Passer domesticus metapopulation to compare lifetime reproductive success (LRS) of resident and immigrant individuals, all with known origin. 3. Lifetime production of recruits by immigrant males was much lower than for resident males, because of shorter life span and lower annual mating success. In contrast, lifetime production of recruits did not differ significantly between immigrant and resident females. 4. Over their lifetime, dispersers contributed fewer recruits to the local population than residents. This shows that immigrant house sparrows have different, sex specific, demographic effects on the population dynamics than residents. PMID- 19674181 TI - Habitat-specific differences in adult survival rates and its links to parental workload and on-nest predation. AB - 1. Adult survival rates strongly affect population growth, but few studies have quantified if and why adult survival differs between breeding habitats. We investigated potential causes of habitat-specific adult survival rates for male and female northern wheatears (Oenanthe oenanthe L.) breeding in Swedish farmland. 2. We used multistate mark-recapture models based on 1263 breeding records between 1993 and 2007 to estimate survival rates based on habitat-type (SHORT vs. TALL ground vegetation) and breeding-success state parameters. We also used breeding-season observations from 2002 to 2007 and an experimental manipulation of ground vegetation height to identify factors influencing adult mortality. 3. Females had lower annual survival than males (0.42 +/- 0.02 vs. 0.50 +/- 0.02); this difference largely resulted from low female survival in TALL habitats because of higher nest-predation risk and the large proportion of adult females being killed on the nest (>20%) during nest predation events. 4. Among successful breeders, both sexes displayed similar survival rates, but survival was lower for breeders in TALL as compared to SHORT habitats (0.43 +/- 0.03 vs. 0.51 +/- 0.02). Experimental manipulation of ground vegetation height, controlling for individual and territory quality (n = 132), suggested the cost of rearing young to be higher in TALL habitats (survival of successful breeders in TALL vs. SHORT; 0.43 +/- 0.11 vs. 0.57 +/- 0.05). 5. Detailed observations of food provisioning behaviour during chick rearing revealed a habitat-related difference in parental workload corresponding to the observed habitat differences in adult survival for successful breeders. Adults breeding in TALL habitats were forced to forage further from the nest relative to SHORT-habitat breeders (mean +/- SE; 69 +/- 10 vs. 21 +/- 2 m), which increased the estimated daily workload for adults in TALL vs. SHORT habitats by c. 20%. 6. On-nest predation and parental workload during chick rearing combine to largely explain habitat specific adult survival rates. The results have implications for our understanding of adult sex ratios, causes of source-sink demography and habitat specific growth rates. Furthermore, it suggests SHORT field margins and other residual habitat elements to be important for the conservation of farmland passerines breeding in cropland plains. PMID- 19674182 TI - Energetic costs of male reproduction in a scramble competition mating system. AB - 1. The assumption that the primary limitations on reproductive success differ between the sexes is inherent in traditional sexual selection theory. Although the energy that can be allocated to reproduction is assumed to be the main limitation to females, the ability to attract and defend oestrous females is assumed to be the primary limitation to males. 2. Estimates of the energetic costs of reproduction in male mammals are, however, limited and have largely been obtained from sexually dimorphic species exhibiting female defence mating systems. These studies often reveal that the energetic cost of male reproduction is similar to or even exceeds that of females, and therefore challenge long-held assumptions of inter-sexual reproductive limitations, but their generality is little known. 3. We coupled measurements of energy expenditure with detailed behavioural observations of reproductive male North American red squirrels (Tamiasciurus hudsonicus). This species displays minimal sexual dimorphism and exhibits a scramble competition mating system, under which sexual selection favours enhanced mate searching effort by males. 4. We conducted the study over 2 years characterized by a substantial variation in upcoming natural food availability and across two study populations that experienced either natural food abundance or an ad libitum food-supplementation to investigate the influence of resource availability on male reproductive energy expenditure. 5. Under natural conditions, mean energy expenditure of males across the 2 years was high, approximating that of females during lactation. Furthermore, in the anticipation of high upcoming natural food availability and resultant offspring survival, expenditure approximately doubled (from 290 +/- 7 to 579 +/- 73 kJ day(-1)). When current food availability (and consequently the density of receptive females) was experimentally elevated, males displayed the highest levels of energy expenditure we recorded (873 +/- 98 kJ day(-1)). 6. Our results provide compelling evidence that the energy available for reproductive allocation places a strong limitation on reproduction in male North American red squirrels and contribute to previous work suggesting that high and limiting energetic costs of male reproduction may be a general feature of mammalian reproduction. PMID- 19674183 TI - Serotypes of Escherichia coli in sudden infant death syndrome. AB - AIM: To examine the diversity of Escherichia coli serotypes found in the intestinal contents of infants who died of Sudden Infant Death Syndrome (SIDS) compared with that in comparison infants. METHODS AND RESULTS: Over the 3-year period, 1989-1991, in South Australia and Victoria (Australia), a total of 687 E. coli isolates from 231 patients with SIDS (348 isolates), 98 infants who had died from other causes (144 isolates) and 160 healthy infants (195 isolates) were studied. The isolates from patients with SIDS were found to represent 119 different serotypes; the isolates from 'other cause' infants represent 97 different serotypes; and the isolates from healthy infants represent 117 different serotypes. The seven common serotypes isolated most frequently from infants with SIDS belonged to those associated with extra-intestinal infections in humans. Compared to healthy infants (6%), these were found in significantly higher proportions among infants who died of other causes (13%, P < 0.05) or infants with SIDS (18.7%, P = 0.0002). CONCLUSIONS: Despite these sources yielding a wide variety of serotypes of E. coli, a pattern of certain potential pathotypes of E. coli being associated with SIDS is apparent. SIGNIFICANCE AND IMPACT OF THE STUDY: While SIDS remains one of the most important diagnoses of postneonatal death, its causes are still unexplained. If E. coli has a role in the pathogenesis of SIDS (as suggested by the pathotypes identified on the basis of serotype), further studies may reveal novel virulence factors that may clarify the role of this bacterium in SIDS. PMID- 19674184 TI - Use of bacterial lipopolysaccharide (LPS) as an immunostimulant for the control of Aeromonas hydrophila infections in rainbow trout Oncorhynchus mykiss (Walbaum). AB - AIMS: To determine the effect of lipopolysaccharide (LPS) for the prevention of infection by Aeromonas hydrophila in rainbow trout (Oncorhynchus mykiss Walbaum) fingerlings. METHODS AND RESULTS: Rainbow trout fingerlings were fed with 0 mg (= controls), 1.875 mg, 3.75 mg, 7.5 mg and 15 mg of LPS per 100 g of commercial feed for 14 days before experimental challenge with A. hydrophila. The results revealed a reduction in mortalities to 5% in the two lowest doses and 15% in the group, which received 15 mg LPS per 100 g of feed, compared with 45% mortalities in the control. LPS exerted a powerful oxidative burst effect and was a potent mediator of phagocytic, lysozyme, bactericidal and antiprotease activities and total protein. However, whereas there were increases in specific growth rate (SGR), feed conversion ratio (FCR) and protein efficiency ratio (PER) in LPS treated fish, the data were not significantly (P > 0.05) different. CONCLUSIONS: LPS was effective at preventing disease caused by A. hydrophila and in stimulating the innate immune response of rainbow trout. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study highlight the role of LPS in fish disease control. PMID- 19674185 TI - Exploitation of a new flagellatropic phage of Erwinia for positive selection of bacterial mutants attenuated in plant virulence: towards phage therapy. AB - AIMS: To isolate and characterize novel bacteriophages for the phytopathogen, Erwinia carotovora ssp. atroseptica (Eca), and to isolate phage-resistant mutants attenuated in virulence. METHODS AND RESULTS: A novel flagellatropic phage was isolated on the potato-rotting bacterial species, Eca, and characterized using electron microscopy and restriction analysis. The phage, named PhiAT1, has an icosahedral head and a long, contractile tail; it belongs to the Myoviridae family. Partial sequencing revealed the presence of genes with homology to those of coliphages T4, T7 and Mu. Phage-resistant transposon mutants of Eca were isolated and studied in vitro for a number of virulence-related phenotypes; only motility was found to be affected. In vivo tuber rotting assays showed that these mutants were attenuated in virulence, presumably because the infection is unable to spread from the initial site of inoculation. CONCLUSIONS: The Eca flagellum can act as a receptor for PhiAT1 infection, and resistant mutants are enriched for motility and virulence defects. SIGNIFICANCE AND IMPACT OF THE STUDY: PhiAT1 is the first reported flagellatropic phage found to infect Eca and has enabled further study of the virulence of this economically important phytopathogen. PMID- 19674186 TI - Impact of Aspergillus section Flavi community structure on the development of lethal levels of aflatoxins in Kenyan maize (Zea mays). AB - AIMS: To evaluate the potential role of fungal community structure in predisposing Kenyan maize to severe aflatoxin contamination by contrasting aflatoxin-producing fungi resident in the region with repeated outbreaks of lethal aflatoxicosis to those in regions without a history of aflatoxicosis. METHODS AND RESULTS: Fungi belonging to Aspergillus section Flavi were isolated from maize samples from three Kenyan provinces between 2004 and 2006. Frequencies of identified strains and aflatoxin-producing abilities were assessed, and the data were analysed by statistical means. Most aflatoxin-producing fungi belonged to Aspergillus flavus. The two major morphotypes of A. flavus varied greatly between provinces, with the S strain dominant in both soil and maize within aflatoxicosis outbreak regions and the L strain dominant in nonoutbreak regions. CONCLUSIONS: Aspergillus community structure is an important factor in the development of aflatoxins in maize in Kenya and, as such, is a major contributor to the development of aflatoxicosis in the Eastern Province. SIGNIFICANCE AND IMPACT OF THE STUDY: Since 1982, deaths caused by aflatoxin-contaminated maize have repeatedly occurred in the Eastern Province of Kenya. The current study characterized an unusual fungal community structure associated with the lethal contamination events. The results will be helpful in developing aflatoxin management practices to prevent future outbreaks in Kenya. PMID- 19674187 TI - The preparation, germination properties and stability of superdormant spores of Bacillus cereus. AB - AIMS: To determine yields, germination and stability of superdormant Bacillus cereus spores. METHODS AND RESULTS: Superdormant B. cereus spores were isolated by germination with high concentrations of inosine or L-alanine in 2-5% yield and did not germinate with high concentrations of either of these germinants, but germinated like starting spores with Ca-DPA, dodecylamine, L-alanine plus inosine or concentrated complete medium. Yields of superdormant spores from germinations with low inosine concentrations were higher, and these spores germinated poorly with low inosine, but relatively normally with high inosine. Yields of superdormant spores were also higher when nonheat-activated spores were germinated. Superdormant spores stored at 4 degrees C slowly recovered some germination capacity, but recovery was slowed significantly at -20 degrees C and 80 degrees C. CONCLUSIONS: Factors that influence levels of superdormant B. cereus spores and the properties of such spores are similar to those in B. megaterium and B. subtilis, suggesting there are common mechanisms involved in superdormancy of Bacillus spores. SIGNIFICANCE: Superdormant spores are a major concern in the food industry, because the presence of such spores precludes decontamination strategies based on triggering spore germination followed by mild killing treatments. Studies of the properties of superdormant spores may suggest ways to eliminate them. PMID- 19674188 TI - Iturin A is the principal inhibitor in the biocontrol activity of Bacillus amyloliquefaciens PPCB004 against postharvest fungal pathogens. AB - AIMS: A Bacillus amyloliquefaciens strain, surviving epiphytically on the surface of fruit, was isolated while searching for naturally occurring biological control agents. This bacterial strain was characterized for its antifungal activity against seven selected fungal postharvest pathogens of citrus. METHODS AND RESULTS: To understand the antifungal activity, seven postharvest fungal pathogens were screened for growth inhibition by B. amyloliquefaciens strain. Assays using B. amyloliquefaciens lipopeptide extracts showed a strong inhibitive activity. The inhibitory effect was observed in abnormal conidial germination and germ tube development when conidia were treated with different lipopeptide extract concentrations. Further analysis using PCR and chromatography confirmed the presence of fengycin, iturin and surfactine, of which iturin A showed the strongest and most common inhibitory effect. The results are supported by site directed mutagenesis analysis, targeted to suppress the biosynthesis of iturin A production. Fruit trials confirmed disease development inhibition when the antagonist was applied 1 day prior to or 1 day after fungal application. CONCLUSIONS: We conclude that the iturin family of lipopeptides are vital in the antagonism of B. amyloliquefaciens against the seven citrus postharvest pathogenic fungi tested. SIGNIFICANCE AND IMPACT OF THE STUDY: We elucidated the principal mechanism used by B. amyloliquefaciens PPCB004 to suppress postharvest disease development on stored fruits. PMID- 19674189 TI - Inactivation and elimination of human enteric viruses by Pacific oysters. AB - AIMS: To investigate the comparative elimination of three different human enterically transmitted viruses [i.e. hepatitis A virus (HAV), norovirus (NoV) and poliovirus (PV)] and inactivation of HAV and PV by Pacific oysters. METHODS AND RESULTS: New Zealand grown Pacific oysters (Crassostrea gigas) were allowed to bioaccumulate HAV, NoV and PV. Samples of oyster gut, faeces and pseudofaeces were then analysed by using real-time RT-PCR to determine the amount of viral RNA and cell culture methods to identify changes in the number of plaque forming units. The results suggest that the majority of the PV present in the oyster gut and oyster faeces is noninfectious, while in contrast, most of the HAV detected in the oyster gut are infectious. Depuration experiments identified a large drop in the count of PV in the gut over a 23-h cleansing period, whereas the levels of HAV and NoV did not significantly decrease. CONCLUSIONS: Human enterically transmitted viruses are eliminated and inactivated at different rates by Pacific oysters. SIGNIFICANCE AND IMPACT OF STUDY: The research presented in this article has implications for risk management techniques that are used to improve the removal of infectious human enteric viruses from bivalve molluscs. PMID- 19674190 TI - Development of HIF-1 inhibitors for cancer therapy. AB - Intratumour hypoxia has long been considered a driving force of tumour progression and a negative prognostic factor in human cancers. The discovery of hypoxia inducible factors (HIFs), which mediate transcriptional responses to changes in oxygen levels, has renewed enthusiasm for the discovery and development of targeted therapies exploiting the hypoxic tumour microenvironment. In spite of an ever increasing number of putative small molecule inhibitors of HIF, only few progress through pre-clinical and early clinical development. In this review, we will focus primarily on: (1) HIF inhibitors that have been more recently described and (2) small molecules targeting HIF that are being tested in early clinical trials or that are already approved for use in patients. A rigorous 'validation' of HIF targeted therapies in relevant pre-clinical models and eventually in pharmacodynamic-based early clinical trials is essential for 'credentialing' HIF-1 as a legitimate target that can be pharmacologically modulated in cancer patients. PMID- 19674192 TI - Transforming growth factor-beta1 elicits Nrf2-mediated antioxidant responses in aortic smooth muscle cells. AB - The anti-inflammatory properties of transforming growth factor-beta(1) (TGF beta(1)) account for its protection against atherosclerotic plaque rupture. This study investigates whether activation of the Nrf2 (nuclear factor erythroid 2 [NF E2]-related factor 2) transcription pathway is involved in TGF-beta(1) mediated induction of the antioxidant enzyme heme oxygenase-1 (HO-1) in smooth muscle cells (SMC). Human aortic smooth muscle cells (HAoSMC) or wild-type and Nrf2 deficient mouse (MAoSMC) aortic SMC were treated with TGF-beta(1) (2.5-10 ng/ml, 0-24 hrs). We report the first evidence that TGF-beta(1) induces Nrf2 mediated HO 1 expression and antioxidant response element activity, which was paralleled by enhanced superoxide production and expression of the NAD(P)H oxidase subunit p22(phox). TGF-beta(1) failed to induce HO-1 expression in MAoSMC derived from Nrf2-deficient mice, and HO-1 induction by TGF-beta(1) in HAoSMC was attenuated by inhibition of extracellular signal regulated kinase or c-jun-N-terminal kinase but not p38 mitogen activated protein kinase. Inhibition of NAD(P)H oxidase or scavenging of superoxide diminished HO-1 induction in response to TGF-beta(1). The oxidative stress agents glucose oxidase (GOx) and diethylmaleate enhanced TGF beta(1) generation and HO-1 expression in HAoSMC, while antagonism of TGF-beta(1) signalling by adenoviral Smad7 overexpression attenuated their induction of HO-1. Pre-treatment of HAoSMC with TGF-beta(1) reduced nuclear translocation of the pro apoptotic mediator p53 elicited by GOx. Our findings demonstrate that Nrf2 is a new target of TGF-beta(1) signalling in the vasculature which may contribute to the atheroprotective properties attributed to this growth factor. PMID- 19674191 TI - Surviving hypoxia by modulation of mRNA translation rate. AB - Cells can survive hypoxia/anoxia by metabolic rate depression, which involves lowering of mRNA translation rates in an ATP-dependent manner. By activating anaerobic ATP production (glycolysis), the inhibitory influence on mRNA translation in hypoxia can be abolished. In severe hypoxia, glycolysis cannot fully restore the ATP demand, thus causing a long-lasting inhibition of global protein synthesis. During moderate hypoxia, fermentative ATP production may maintain normal ATP levels. However, an activation of hypoxia tolerance mechanisms, including specific mRNA translation, also takes place. The latter may be attributed to oxygen-dependent (but not ATP dependent) processes such as the activation of the hypoxia-inducible factor cascade. In summary, hypoxia-induced decline in cellular ATP level can be counteracted by suppression of global mRNA translation rate. Sustained protein synthesis seems to be attributed to the activation of specific mRNA translation under long-term hypoxic conditions. PMID- 19674194 TI - The role of traumatic event history in non-medical use of prescription drugs among a nationally representative sample of US adolescents. AB - BACKGROUND: Building on previous research with adolescents that examined demographic variables and other forms of substance abuse in relation to non medical use of prescription drugs (NMUPD), the current study examined potentially traumatic events, depression, posttraumatic stress disorder (PTSD), other substance use, and delinquent behavior as potential correlates of past-year non medical use of prescription drugs. METHOD: A nationally representative sample of 3,614 non-institutionalized, civilian, English-speaking adolescents (aged 12-17 years) residing in households with a telephone was selected. Demographic characteristics, traumatic event history, mental health, and substance abuse variables were assessed. NMUPD was assessed by asking if, in the past year, participants had used a prescription drug in a non-medical manner. Multivariable logistic regressions were conducted for each theoretically derived predictor set. Significant predictors from each set were then entered into a final multivariable logistic regression to determine significant predictors of past-year NMUPD. RESULTS: NMUPD was endorsed by 6.7% of the sample (n = 242). The final multivariable model showed that lifetime history of delinquent behavior, other forms of substance use/abuse, history of witnessed violence, and lifetime history of PTSD were significantly associated with increased likelihood of NMUPD. CONCLUSIONS: Risk reduction efforts targeting NMUPD among adolescents who have witnessed significant violence, endorsed abuse of other substances and delinquent behavior, and/or endorsed PTSD are warranted. Interventions for adolescents with history of violence exposure or PTSD, or those adjudicated for delinquent behavior, should include treatment or prevention modules that specifically address NMUPD. PMID- 19674195 TI - Maternal pre-pregnancy obesity and risk for inattention and negative emotionality in children. AB - OBJECTIVE: This study aimed to replicate and extend previous work showing an association between maternal pre-pregnancy adiposity and risk for attention deficit hyperactivity disorder (ADHD) symptoms in children. METHODS: A Swedish population-based prospective pregnancy-offspring cohort was followed up when children were 5 years old (N = 1,714). Mothers and kindergarten teachers rated children's ADHD symptoms, presence and duration of problems, and emotionality. Dichotomized outcomes examined difficulties of clinical relevance (top 15% of the distribution). Analyses adjusted for pregnancy (maternal smoking, depressive symptoms, life events, education, age, family structure), birth outcomes (birth weight, gestational age, infant sex) and concurrent variables (family structure, maternal depressive symptoms, parental ADHD symptoms, and child overweight) in an attempt to rule out confounding. RESULTS: Maternal pre-pregnancy overweight and obesity predicted high inattention symptom scores and obesity was associated with a two-fold increase in risk of difficulties with emotion intensity and emotion regulation according to teacher reports. Means of maternal ratings were unrelated to pre-pregnancy body mass index (BMI). Presence and duration of problems were associated with both maternal over and underweight according to teachers. CONCLUSIONS: Despite discrepancies between maternal and teacher reports, these results provide further evidence that maternal pre-pregnancy overweight and obesity are associated with child inattention symptoms and extend previous work by establishing a link between obesity and emotional difficulties. Maternal adiposity at the time of conception may be instrumental in programming child mental health, as prenatal brain development depends on maternal energy supply. Possible mechanisms include disturbed maternal metabolic function. If maternal pre-pregnancy obesity is a causal risk factor, the potential for prevention is great. PMID- 19674193 TI - Blockade of GRP78 sensitizes breast cancer cells to microtubules-interfering agents that induce the unfolded protein response. AB - Taxane and vinblastine represent two classes of microtubules-targeted agents for cancer chemotherapy. Although taxol and vinblastine are widely used for cancer treatment, resistance to these agents is frequently encountered in the clinic. An ongoing question has been what mechanisms are involved in the resistance of tumour cells to microtubules-targeted agents or how the clinical effectiveness can be improved. There is increasing evidence that microtubules interact with the endoplasmic reticulum (ER). Here, we have shown that taxol and vinblastine induce multiple arms of the ER stress response, including up-regulation of glucose regulated protein 78 (GRP78) expression, X-box binding protein 1 splicing and eukaryotic initiation factor 2alpha phosphorylation. Abrogation of GRP78 induction sensitizes breast cancer cells to taxol and vinblastine. Treatment with (-)-epigallocatechin gallate (EGCG), a known GRP78 inhibitor, synergistically promotes taxol- and vinblastine-induced cell death. GRP78 knockdown or EGCG potentiates taxol- and vinblastine-induced activation of pro-apoptosis arms of the ER stress response, such as JNK phosphorylation, caspase-7 and PARP cleavage. Inhibition of JNK and caspase-7 abrogates EGCG sensitization of breast cancer cells to taxol and vinblastine. We conclude that induction of the unfolded protein response represents a novel mechanism underlying the efficacy and resistance to microtubules-targeted agents. Combination of compounds capable of suppressing GRP78 might be a novel approach for improving the effectiveness of microtubules-targeted chemotherapy. PMID- 19674196 TI - Electrophysiological indices of abnormal error-processing in adolescents with attention deficit hyperactivity disorder (ADHD). AB - BACKGROUND: Impaired cognitive control has been frequently observed in children and young people with attention deficit hyperactivity disorder (ADHD) and might underlie the excessive hyperactivity and impulsivity in this population. We investigated behavioural and electrophysiological indices relevant to one domain of cognitive control; namely error processing. METHODS: Adolescents aged 14 to 17 with ADHD (n = 23) and a typically developing control group (HC; n = 19) performed a visual go/no-go task. Electro-encephalography (EEG) data were collected simultaneously and response-locked error trials were averaged to derive two event-related potentials, the error-related negativity (ERN) and error positivity (Pe). Evoked theta power and inter-trial phase coherence (ITC) were measured in two time windows ('early' and 'late') equivalent to those used for detection of the ERN and Pe. RESULTS: Analysis revealed normal ERN amplitude and a statistical trend for smaller Pe amplitude at a fronto-central electrode site in the ADHD group. The group also showed significant reductions in late evoked theta power and early and late theta ITC. Relationships between behavioural measures and ITC were different between groups, particularly for post-error slowing, a measure of strategic response adjustment on trials immediately following an error. CONCLUSIONS: The results reveal abnormalities in behavioural and electrophysiological indices of error processing in adolescents with ADHD and suggest that ITC is more sensitive than traditional ERP measures to error processing abnormalities. PMID- 19674197 TI - Paucity of intraepidermal FoxP3-positive T cells in cutaneous T-cell lymphoma in contrast with spongiotic and lichenoid dermatitis. AB - BACKGROUND: FoxP3 is the most specific available marker for regulatory T cells (Tregs). Tumor-associated FoxP3-positive Tregs have been identified in various neoplasms, including cutaneous T-cell lymphoma (CTCL). FoxP3 expression in CTCL varies across groups; few studies have compared CTCL with inflammatory conditions. METHODS: Lesional skin biopsies from 20 patients with CTCL [13 mycosis fungoides (MF); 7 Sezary syndrome (SS)] and 22 with inflammatory dermatoses (11 spongiotic; 11 lichenoid or interface) were examined for FoxP3 expression by immunohistochemistry. Epidermal FoxP3-positive lymphocytes were counted as a percentage of the total epidermal CD3-positive T-cell population. RESULTS: FoxP3-positive T cells composed the minority of infiltrate in all major categories. Lower numbers of epidermal FoxP3-positive T cells were observed in CTCL, particularly MF, than in inflammatory dermatoses (P < .001). CTCL neoplastic T cells did not express FoxP3. CONCLUSION: FoxP3-positive T cells are less frequently encountered in MF than in inflammatory dermatoses. FoxP3-positive T cells occur in higher proportions in the dermis than in the epidermis and probably correlate with coexisting inflammatory components. CTCL neoplastic cells do not typically express a Treg phenotype and are associated with low numbers of FoxP3-positive Tregs in the infiltrate. FoxP3 expression by immunohistochemistry may aid histologic evaluation of these conditions. PMID- 19674198 TI - Matrix metalloproteinase-26 is present more frequently in squamous cell carcinomas of immunosuppressed compared with immunocompetent patients. AB - BACKGROUND: Skin cancers are the most frequent malignancies in organ transplant recipients (OTRs). Squamous cell carcinomas (SCCs) occur 65-250 times more frequently in OTRs and tend to be aggressive in behavior. Because matrix metalloproteinases (MMPs) have a central role in tumorigenesis and invasion, we investigated the epithelial and stromal MMP and tissue inhibitor of MMP (TIMP) expression profile in SCCs of immunosuppressed (IS) compared with immunocompetent (IC) patients to determine if differences could explain the more aggressive behavior of SCCs in OTRs. METHODS: Matched pairs from 20 SCCs of IS and IC patients were studied using immunohistochemistry for MMP-1, MMP-7, MMP-8, MMP-9, MMP-13 and MMP-26 and TIMP-1 and TIMP-3. RESULTS: Among all MMPs studied, only staining for MMP-26 was significantly more intense in cancer cells of the post transplant group compared with the IC group (p = 0.01), whereas MMP-9 expression was more abundant in stromal macrophages surrounding SCCs of IC patients (p = 0.02). MMP-26 expression in cancer cells (p = 0.04) and that of MMP-9 in neutrophils (p = 0.005) were more abundant in SCCs of patients using cyclosporine. CONCLUSIONS: We conclude that MMP-26 and MMP-9 may contribute to the more aggressive behavior of SCCs in OTRs. PMID- 19674199 TI - Fibrofolliculoma/trichodiscoma and fibrous papule (perifollicular fibroma/angiofibroma): a revaluation of the histopathological and immunohistochemical features. AB - BACKGROUND: The multiple facial lesions of fibrofolliculoma (FF)/trichodiscoma (TD) and those of fibrous papule (FP; perifollicular fibroma/angiofibroma, AF) are characteristic of Birt-Hogg-Dube (BHD) syndrome and tuberous sclerosis, respectively. However, there was a recently reported case of BHD syndrome with multiple facial FP lesions and a case of tuberous sclerosis, in which one FF lesion was included among the multiple facial FPs. METHODS: The histopathological and immunohistochemical features of FF/TD and FP lesions were revaluated to study the relationship between the two. This investigation evaluated 20 FF/TD lesions including two cases of BHD syndrome and 35 FP lesions including three cases of tuberous sclerosis. RESULTS: There are common histopathological features in the two lesions, such as the follicular and perifollicular elements, an angiofibromatous element and stellate fibrocytes. The similar immunohistochemical features between the two lesions included the expression patterns of CD34, factor XIIIa, nestin and c-kit in the stromal cells as well as abnormal CK15 expression in the hair follicles. CONCLUSIONS: Both FF/TD and FP are hamartomas composed of perifollicular or interfollicular connective tissue and a hair follicular epithelial component, which may be caused by an abnormal functioning of the hair follicle bulge cells. PMID- 19674200 TI - Elastic fiber pattern in scleroderma/morphea. AB - BACKGROUND: Scleroderma/morphea is characterized by expansion of the dermis with thickened collagen bundles and loss of CD34(+) dermal dendrocytes. Variable elastic fiber changes have been described, but to our knowledge, no systematic study of the elastic fiber pattern correlated with CD34 expression has been reported. METHODS: To better define the typical elastic fiber morphology, we examined seven cases of normal skin and 28 cases of scleroderma/morphea ranging from inflammatory to sclerosing stages. All but four biopsies were submitted with a clinical impression of either scleroderma or morphea. CD34 immunohistochemistry was performed on 26 biopsies with available tissue. RESULTS: Elastic van Gieson stain showed preservation of elastic fibers in all cases. In addition, straightening with parallel orientation and compression between thickened collagen bundles was frequently present and was graded as limited in 46% and diffuse in 54% of cases. The extent of elastic fiber alteration correlated with the degree of sclerosis. A variable loss of CD34(+) dermal dendritic cells was seen in all cases. CONCLUSION: This study confirms the preservation and frequent presence of parallel, straightened and compressed elastic fibers in scleroderma/morphea and suggests that the elastic fiber pattern, in addition to CD34 immunohistochemistry, may serve as a useful diagnostic adjunct. PMID- 19674201 TI - Skin biopsy is helpful for the diagnosis of incontinentia pigmenti at late stage (IV): a series of 26 cutaneous biopsies. AB - BACKGROUND: Hypochromic streaks can be the only cutaneous sign of incontinentia pigmenti (IP) in adulthood (stage IV). Discovery of such lesions in an adult female with no family history of IP is essential for appropriate genetic counselling. OBJECTIVE: To describe and to validate the histological features of residual skin lesions in adult IP. METHODS: The analysis and comparison of skin biopsies of 26 women affected with molecularly confirmed IP. RESULTS: Most biopsies showed slight atrophy and some scattered apoptotic cells in the epidermis, epidermal hypopigmentation and reduced melanocyte number. The dermis appeared thickened and homogeneous and revealed a complete absence of hair follicles (23/26) and sweat glands (22/26). There was no melanin incontinence or inflammatory cells, and the elastic network was normal. CONCLUSION: These features lead unequivocally to the diagnosis of a stage IV IP skin lesion. Consequently, histology is a major confirmatory criterion for diagnoses of these mild clinical forms of IP. It is therefore a useful tool in genetic counselling and prenatal diagnosis. Moreover, the observations described here may contribute to understanding the physiopathology of the late stages of IP. PMID- 19674202 TI - Retiform hemangioendothelioma: presentation of a case expressing D2-40. AB - Retiform hemangioendothelioma (RH) is a low-grade angiosarcoma with low metastatic risk, usually occurring as a single lesion on the trunk or extremity in middle-aged adults. Histopathology shows a distinctive pattern with arborizing blood vessels arranged in a retiform pattern (similar to rete testis tissue) and focal papillae with fibrosclerotic (hyaline) cores. The blood vessels are lined by comparatively monomorphic endothelial cells, frequently presenting a hobnail pattern. We report a case of RH presenting as an indolent brownish plaque on the back of a 17-year-old male. Surgical resection and sentinel lymph node biopsy showed no evidence of metastasis. In contrast to the recent literature, this case of RH showed positivity for D2-40, a marker of lymphatic endothelium. We also report ultrastructural findings for this case of RH. PMID- 19674204 TI - Acantholytic dyskeratotic epidermal nevus with eccrine differentiation: a case report and review of literature. AB - Terminology regarding classification of benign lesions with prominent eccrine differentiation can be confusing as these lesions can have overlapping clinical and histologic characteristics. In this report, we examine a case and review of the literature to suggest that these entities may be better classified as a spectrum of benign lesions with overlapping features rather than distinct entities. We describe a case of an acantholytic dyskeratotic epidermal nevus with eccrine differentiation on the back of a 2-year-old patient. We then discuss the classic clinical and histologic presentations of eccrine nevi and epidermal nevi with eccrine differentiation as they relate to each other and to our case. PMID- 19674203 TI - Pigmented extramammary Paget's disease of the axilla mimicking melanoma: case report and review of the literature. AB - Pigmented Paget's disease is a rare variant that is often confused clinically and histologically with melanoma in situ. Herein, we describe a case of pigmented extramammary Paget's disease involving the axilla of a 79-year-old white male thought initially to represent malignant melanoma clinically and histologically. Review of the literature reveals that pigmented variant of Paget's disease, either mammary or extramammary, could be initially misdiagnosed as melanoma unless this entity is considered in the differential diagnosis, and additional confirmatory studies are performed. PMID- 19674205 TI - No indications for HPV involvement in the hypertrophic skin lesions of a Darier disease case without ATP2A2 gene mutations. AB - Darier disease (DD) is a relatively common genodermatosis characterized by impaired differentiation and abnormal cell-to-cell adhesion. Haploinsufficiency of the ATP2A2 gene product, sarco/endoplasmic reticulum Ca(2+) ATPase isoform 2 (SERCA2), is the underlying cause of most cases. Although DD may have a papillomatous appearance, few and controversial results have been reported about the role of human papillomavirus (HPV) in this disease. The aim of this study was to determine a possible correlation between development of hypertrophic lesions in DD and infection by HPV. We report the case of an 84-year-old woman with a hypertrophic DD variant that has been successfully treated with oral retinoids. HPV analysis for a broad spectrum of cutaneous and mucocutaneous genotypes was performed on surgical specimens obtained from the cutaneous lesions and snap frozen plucked eyebrows. Genetic analysis of the ATP2A2 gene did not detect any mutations. Epidermal expression of SERCA2b was shown by immunohistochemistry. We describe a patient with DD lacking mutations of the ATP2A2 gene, but with reduced SERCA2b expression in the epidermal keratinocytes. The results obtained by polymerase chain reaction (PCR) genotyping, quantitative real-time PCR, and in situ hybridization indicate that HPV replication was very low and suggest no direct role of the virus in the development of the disease. PMID- 19674206 TI - Cytological atypia does not equal malignancy: an old but unappreciated truth. PMID- 19674207 TI - When a negative is a positive! PMID- 19674208 TI - Neutrophilic panniculitis associated with myelodysplastic syndrome with abnormal nuclear forms. PMID- 19674209 TI - De novo intraepidermal epithelioid melanocytic dysplasia, a unique observation in a complex discipline. PMID- 19674210 TI - Defining the hair follicle stem cell (Part I). PMID- 19674212 TI - What do we know of post-thrombotic syndrome? Current status of post-thrombotic syndrome in adults. AB - The post-thrombotic syndrome (PTS) is a common but not completely understood and rarely studied sequela of an acute deep vein thrombosis. The influence of several risk factors on the incidence or severity of PTS is controversial. The therapeutic options for PTS are still limited. Appropriate medical compression stockings can reduce the incidence (-50 %) as well as the severity of PTS. In the case of severe, symptomatic PTS, they should be worn lifelong. As PTS is a chronic and often progressive disease, patients need regular clinical follow-up and encouragement to continuously wear their compression therapy. New strategies for treating PTS are needed. PMID- 19674213 TI - Mortality of Parkinson's disease by Hoehn-Yahr stage from community-based and clinic series [Keelung Community-based Integrated Screening (KCIS) no. 17)]. AB - PURPOSE: We aimed to quantify the mortality reduction by which the early detection of Parkinson's disease (PD) within a community-based study could reduce the number of advanced cases. METHODS: Data used in this study were derived from two community-based surveys and from a clinical series of PD cases identified from a medical centre. The cumulative survival by Hoehn-Yahr (H-Y) scale was estimated and the mortality reduction derived from a community-based survey was predicted. RESULTS: A total of 117 PD patients were detected through two community-based approaches. By comparing the H-Y stage distribution of screen detected cases with those from the clinical series, a 22.5% excess in the number of early PD (H-Y stage I and stage II) were identified with screening. The risk ratios of being H-Y stage III or severe for community-based detected cases versus clinical series were 0.49 (95% confidence interval: 0.30-0.78). The total death rate adjusted by H-Y stage distribution was 21% and 28% for cases from community and clinical series, respectively. CONCLUSIONS: The present study revealed that early detection of PD through a community-based survey may reduce 51% incidence of stage III or more severe PD at diagnosis, leading to a 25% reduction in mortality. PMID- 19674214 TI - Waiting list management: priority criteria or first-in first-out? A case for total joint replacement. AB - BACKGROUND: Total joint replacements are interventions with large waiting times from indication to the surgery management. These patients can be managed in two ways; first-in first-out or through a priority tool. The aim of this study was to compare real time on waiting list (TWL) with a priority criteria score, developed by our team, in patients awaiting joint replacement due to osteoarthritis. METHODS: Consecutive patients placed on waiting list were eligible. Patients fulfilled a questionnaire which included items of our priority tool and the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) specific questionnaire. Other priority items were extracted from the clinical history. The priority tool gives a score from 0 to 100 points, and three categories (urgent, preferent and ordinary). We studied the differences among categories and TWL by means of one-way analysis of variance. Correlational analysis was used to evaluate association among priority score and TWL and WOMAC baseline and gains at 6 months with priority score and TWL. RESULTS: We have studied 684 patients. Women represented 62% of sample. The mean age was 70 years. There were not association between the categories of priority score and TWL (P = 0.12). The rho correlation coefficient between TWL and priority score was -0.11. Among baseline WOMAC scores and priority score, the rho coefficients were 0.79, 0.7 and 0.52 with function, pain and stiffness dimensions, respectively. There were differences in the mean scores of WOMAC dimensions according to the three priority categories (P < 0.001) but no with TWL categories. Data of gains in both health-related quality of life dimensions at 6 months were similar, with differences according to priority categories but no regarding TWL. CONCLUSIONS: The results of the study support the necessity of implementing a prioritization system instead of the actual system if we want to manage the waiting list for joint replacement with clinical equity. PMID- 19674215 TI - Training health care providers in the treatment of tobacco use and dependence: pre- and post-training results. AB - RATIONALE: Health care providers have an extended reach into the population of tobacco users. Increasing the number and variety of health care providers that deliver the evidence-based, brief interventions for tobacco use prescribed by the Public Health Service Clinical Practice Guideline is likely to result in more tobacco users exposed to evidence-based treatments and more successful quit attempts. Effective training is key to increasing provider performance and proficiency in this regard. METHOD: A 1-hour didactic training was delivered to 1286 health care providers (185 physicians, 359 nurses, 75 dental providers and 667 other health-related professions). Pre- and post-training tests assessed provider attitudes, knowledge and behaviours. Paired samples t-tests were used to compare pre- and post-test results. Analysis of variance was used to test for significant differences among professional groups. RESULTS: Prior to training, physicians engaged in more interventions and reported more knowledge and more positive attitudes towards treating tobacco use than the other professions. Post training, differences among physicians, nurses and dental providers were minimal. All professions reported significantly more knowledge and more positive attitudes on nearly all measures. CONCLUSIONS: A large potential benefit can be garnered from a brief, targeted, 1-hour training in the brief, evidence-based interventions for treating tobacco use and dependence. Increases in perceived knowledge and positive attitudes towards treatment among the professional groups suggest that trainees will perform interventions at higher frequency post training. Overall gains were highest for dental providers and nurses. PMID- 19674216 TI - Mental health consumers' perceptions of receiving recovery-focused services. AB - RATIONALE, AIMS AND OBJECTIVES: This study examines the experiences of mental health service consumers engaged in various recovery-focused support practices as well as examining consumer valuing of these activities. METHOD: A self-report questionnaire was developed drawing on key aspects of the Collaborative Recovery Model (CRM) (responsibility, collaboration, autonomy, motivation, needs, goals, homework). Ninety-two adult consumers from metropolitan, regional and rural non government organizations and public mental health services in eastern Australian states completed the questionnaire. RESULTS: Consumers using services provided by CRM-trained workers identified significant changes to service delivery in relation to frequency with which they were encouraged to take responsibility for their recovery, degree to which they collaborated with staff and the extent to which they were encouraged to complete homework activities to assist them to achieve their goals, when compared with consumers using traditional services. The key aspects of the CRM were valued by consumers. No differences were found in terms of overall ratings of clinician helpfulness in assisting recovery between the two groups. CONCLUSIONS: Consumers are able to perceive recovery-focused service changes. Although preliminary, this is a significant step towards assessing the operationalization of recovery principles from the consumer's perspective. PMID- 19674217 TI - A multi-intervention approach on drug therapy can lead to a more appropriate drug use in the elderly. LIMM-Landskrona Integrated Medicines Management. AB - RATIONALE, AIMS AND OBJECTIVES: To evaluate if an integrated medicines management can lead to a more appropriate drug use in elderly inpatients. METHOD: The study was an intervention study at a department of internal medicine in southern Sweden. During the intervention period pharmacists took part in the daily work at the wards. Systematic interventions aiming to identify, solve and prevent drug related problems (DRPs) were performed during the patient's hospital stay by multidisciplinary teams consisting of physicians, nurses and pharmacists. DRPs identified by the pharmacist were put forward to the care team and discussed. Medication Appropriateness Index (MAI) was used to evaluate the appropriateness in the patients' drug treatment at admission, discharge and 2 weeks after discharge. In total 43 patients were included, 28 patients in the intervention group and 25 patients in the group which was used as control. RESULTS: For the intervention group there was a significant decrease in the number of inappropriate drugs compared with the control group (P = 0.049). Indication, duration and expenses were the MAI-dimensions with most inappropriate ratings, and the drugs with most inappropriate ratings were anxiolytics, hypnotics and sedatives. CONCLUSION: This kind of systematic approach on drug therapy can result in a more appropriate drug use in the elderly. PMID- 19674218 TI - Evaluating the effectiveness of a deep-vein thrombosis prophylaxis protocol in orthopaedics and traumatology. AB - RATIONALE, AIMS AND OBJECTIVES: To evaluate the effectiveness of the deep-vein thrombosis (DVT) prophylaxis protocol for adult patients in a general orthopaedics and traumatology clinic. METHOD: We followed the DVT prophylaxis protocol in 1326 (776 female, 550 male) of 2114 adult patients admitted to the Department of Orthopaedics and Traumatology in Goztepe Research and Training Hospital. They were followed for symptomatic DVT and possible complications of low-molecular-weight heparin (LMWH) therapy. A Doppler ultrasonography (US) was performed when DVT was suspected. The medical information treatment protocols of DVT patients were recorded. RESULTS: Doppler US was performed in 58 patients with suspected DVT. Six of these patients were diagnosed with DVT. The side effects of LMWH were upper gastrointestinal bleeding (0.5%), widespread ecchymosis of the extremities (1.9%) and heparin-induced thrombocytopenia (0.16%). CONCLUSION: Symptomatic DVT occurrences were similar to those in medical literature; however, there were fewer side effects of LMWH than reported in literature. PMID- 19674220 TI - Explicit versus implicit review to explore combination antipsychotic prescribing. AB - OBJECTIVE: To use structured implicit review following large-scale explicit audit of antipsychotic polyprescribing to: (1) determine the true rate of antipsychotic polytherapy that deviated from best practice for schizophrenia treatment; and (2) assess whether explicit antipsychotic polytherapy criterion was appropriate for identifying patients at risk for medication problems and assessing quality of care. METHODS: Antipsychotic prescribing was reviewed for outpatients in four public health services in Auckland, New Zealand on 31 October 2004 (T1). Schizophrenia patients in one service (n = 794) prescribed antipsychotic polytherapy (n = 84, 10.6%) were followed up 10 months later (T2). Historical medication summaries were prepared for those remaining on polytherapy, including diagnosis, clinical problems and treatment plan. Criteria for structured implicit review and rating form for quality of antipsychotic management were piloted. All medication summaries were independently rated by two reviewers, and a third independent rater reviewed summaries where disagreement was found. RESULTS: Forty nine patients remained on long-term polytherapy at T2 (6.2% of original population). All but two cases included a second-generation antipsychotic. At T2, average polytherapy duration was 35.8 months, and average antipsychotic dose was 699 mg day(-1) chlorpromazine equivalents. Two raters achieved agreement for 24/49 summaries, and the remaining 25 were rated independently by a third reviewer. Consensus agreement of antipsychotic management (by two raters) was reached for 44/49 cases (89.8%). Polytherapy was rated 'well-justified' in 32.7%, 'some justification' in 10.2% and 'lacked justification' in 46.9% cases. The final rate of polytherapy deviating from best practice reduced from 10.6% to 3.5% when short-term polytherapy was excluded, and details of the clinical situation and care plan were included in implicit review. CONCLUSIONS: Audit of prescribing in routine practice using explicit guideline-based criteria may be a useful baseline performance indicator. It does not provide an accurate measurement of quality of care because it overestimates the deviation rate from good practice. It may also identify complex patients at risk for poor treatment outcomes who may benefit from structured treatment review. PMID- 19674219 TI - Validation of priority criteria for cataract extraction. AB - RATIONALE, AIMS AND OBJECTIVES: Given the increasing prevalence of cataract and demand for cataract extraction surgery, patients must often wait to undergo this procedure. We validated a previously developed priority scoring system in terms of clinical variables, pre-intervention health status, appropriateness of surgery and gain in visual acuity (VA) and health-related quality of life (HRQoL). METHODS: Explicit prioritization criteria for cataract extraction created by a variation of the Research and Development (RAND) and University of California Los Angeles appropriateness methodology were retrospectively applied to a prospective cohort of 5257 patients on waiting lists to undergo cataract by phacoemulsification at 17 hospitals in Spain. Demographic data, clinical data and data related to surgical technique were collected by trained ophthalmologists. Patients were evaluated by their ophthalmologist before the intervention and 6 weeks afterward. They also completed, by mail, the Visual Function Index (VF-14) before the intervention and 3 months afterward. RESULTS: High-priority patients experienced greater improvement in VA and HRQoL than those classified as intermediate or low-priority (P < 0.0001), even after adjusting by VA and the VF 14 score at baseline. The time to intervention was the same for high-priority patients as it was for intermediate and low-priority patients. CONCLUSIONS: The priority score we developed identified patients most likely to experience the greatest improvements from cataract extraction. Use of this tool could provide a fairer and more rational way to prioritize patients for cataract extraction. PMID- 19674221 TI - Chronic Care Team Profile: a brief tool to measure the structure and function of chronic care teams in general practice. AB - AIM: At a time when workforce shortages in general practices are leading to greater role substitution and skill-mix diversification, and the demand on general practices for chronic disease care is increasing, the structure and function of the general practice team is taking on heightened importance. To assist general practices and the organizations supporting them to assess the effectiveness of their chronic care teamworking, we developed an interview tool, the Chronic Care Team Profile (CCTP), to measure the structure and function of teams in general practice. This paper describes its properties and potential use. METHOD: An initial pool of items was derived from guidelines of best-practice for chronic disease care and performance standards for general practices. The items covered staffing, skill-mix, job descriptions and roles, training, protocols and procedures within the practice. The 41-item pool was factor analysed, retained items were measured for internal consistency and the reduced instrument's face, content and construct validity were evaluated. RESULTS: A three-factor solution corresponding to non-general practitioner staff roles in chronic care, administrative functions and management structures provided the best fit to the data and explained 45% of the variance in the CCTP. Further analyses suggested that the CCTP is reliable, valid and has some utility. DISCUSSION: The CCTP measures aspects of the structure and function of general practices which are independent of team processes. It is associated with the job satisfaction of general practice staff and the quality of care provided to patients with chronic illnesses. As such, the CCTP offers a simple and useful tool for general practices to assess their teamworking in chronic disease care. PMID- 19674222 TI - Improvement in health-related quality of life, independent of fasting glucose concentration, via insulin pen device in diabetic patients. AB - RATIONALE AND AIMS: Using an insulin pen may improve the quality of life (QOL) for diabetic individuals. However, glycemic control and its relationship to better QOL are controversial. The aim of this study was to determine the relationship between health-related QOL and glycemic control in diabetic patients using insulin pen. METHODS: All of the participants were diabetic patients receiving insulin therapy by syringe injection for longer than 1 month. One group of enrolled subjects changed over to use of insulin pen for 12 weeks, while the other group (age-matched control subjects) continued to use syringe for insulin injection during the same period. Fasting plasma glucose, HbA1c and a 36-Item Short-Form Health Survey (SF-36) were assessed in both groups before and after the 12-week study. RESULTS: A total of 32 subjects in the insulin pen group and 33 subjects in the syringe group completed the assessment. In comparison with baseline, fasting glucose significantly decreased in the insulin pen group (-57 +/- 14 mg dL(-1), P < 0.001), and the reduction was significantly greater than that in the syringe group (P = 0.003). The summary scale of physical components but not mental components in the SF-36 was significantly higher in the insulin pen group than in the syringe group (P = 0.037). This improvement was independent of the change in fasting glucose. CONCLUSIONS: Using insulin pen for insulin injection improved glycemic control and health-related QOL in diabetic patients. The better functional health status as a result of physical improvement was independent of the glycemic control. PMID- 19674223 TI - An audit of health education services within UK hospitals. AB - RATIONALE, AIMS AND OBJECTIVE: UK public health policy requires hospitals to deliver health promotion services to patients for healthy lifestyles (i.e. health education), but there are currently few data on the health education delivered within hospitals. This audit aimed to collect data on the routine health education activities delivered to hospitalized patients to assess whether the following standards were met: 100% of hospitalized patients screened for smoking, alcohol use and obesity, 70% of smokers offered health education for smoking cessation and 50% of patients identified as misusing alcohol, obese, consuming an unhealthy diet and/or physically inactive delivered the appropriate health education. METHODS: An audit of data contained in hospitalized patients' written medical case notes for evidence that the above standards were met. Nine hospitals in Greater Manchester in England participated. RESULTS: Four hospitals screened all patients for smoking. None of the hospitals met the standards for screening alcohol or obesity. For health education delivery, all hospitals met the standard for diet, four for alcohol misuse and four for physical activity. None of the hospitals met the standards for smoking or obesity. CONCLUSIONS: Improvements in practice for screening of alcohol and obesity are required. While some hospitals appeared to meet standards for health education delivery for alcohol, diet and physical activity, given the poor screening procedures for these risk factors, we can not conclude that health education delivery was adequate. PMID- 19674224 TI - A structured questionnaire to assess patient compliance and beliefs about medicines taking into account the ordered categorical structure of data. AB - RATIONALE, AIMS AND OBJECTIVE: The objectives were to describe and evaluate the structured medication questionnaire and to improve data handling of results from the Morisky four-item scale for patient compliance and Beliefs about Medicines Questionnaire-specific (BMQ-specific). METHODS: A questionnaire was developed with the purpose of being used when identifying medication errors and assessing patient compliance to and beliefs about medicines. RESULTS: A majority of the respondents (62%; CI 45-77%) had at least one medication error. Assuming that all items are equally important in the Morisky four-item scale we presented four alternative ways to create a unidimensional global scale. A two-dimensional global scale was also constructed. The results from the BMQ-specific were presented in different ways, all taking into account that the scale has ordered verbal categories: at the level addressing each specific question, at the sub scales 'concern' and 'necessity' level and at the global level. CONCLUSIONS: The structured medication questionnaire can be used in daily practice as a tool to identify drug-related problems. The choice of how to use and present data from those scales in research depends on patient characteristics and how discriminating one would like the scales to be. PMID- 19674225 TI - Do physical therapists change their beliefs, attitudes, knowledge, skills and behaviour after a biopsychosocially orientated university course? AB - AIM: The aim of this study is to examine the effects of an 8-day university-based training course, aimed at identifying and addressing psychosocial prognostic factors during physiotherapy treatment, in shifting therapists towards a more biopsychosocial orientation as measured by changes in beliefs/attitudes, knowledge, skills and behaviour. METHOD: We combined a randomized controlled trail with a pre-post design. Forty-two physiotherapists applied for a university accredited training course designed to enhance knowledge and management of psychosocial factors in their practice with patients suffering from musculoskeletal pain. The course participants were randomized either to receiving the course or to a waiting list for training. Attitudes and beliefs towards, and knowledge of psychosocial factors, patient vignettes and a video of an imaginary patient were tested before and after training. The patients of the course participants were asked to fill out a questionnaire with background questions at treatment start. The patients also received a questionnaire about the physical therapists' behaviour and patient satisfaction 6 weeks after treatment start. RESULTS: The results show that physical therapists' attitudes and believes became more biopsychosocially and less biomedically orientated, they were less convinced that pain justifies disability and limitation of activities, and their knowledge and skills on psychosocial risk factors increased after a university-accredited training course. Yet despite these changes their patients perceived their practice behaviour before and after the course as similar and were equally satisfied with their treatment and treatment result. CONCLUSION: A course, which enhanced biopsychosocial attitudes and beliefs, as well as increased such knowledge and skills did not change the way patients perceived their physical therapists. A future question is whether it improves patient outcome. PMID- 19674226 TI - Questionnaire instrument to assess knowledge of chronic kidney disease clinical practice guidelines among internal medicine residents. AB - INTRODUCTION: The National Kidney Foundation published Kidney Disease Outcomes Quality Initiative guidelines that recommend early detection and management of chronic kidney disease (CKD) and timely referral to a nephrologist. Many patients with CKD are seen by primary care doctors who maybe less experienced than a nephrologist to offer optimal early CKD care. It is not known whether postgraduate training adequately prepares a future internist in CKD management. METHODS: We developed a 15-item questionnaire instrument to assess knowledge of CKD guidelines among internal medicine residents in USA using an online survey programme. We studied the validity and reliability measures of our instrument. RESULTS: The survey was completed by 166 PGY1 (postgraduate year one), 187 PGY2 and 126 PGY3. The questionnaire tested various aspects of CKD including definition, classification, identification of risk factors, laboratory evaluation, development of clinical action plan, identification of complications, anaemia and bone and mineral disorder, referral to a nephrologist and medication use. Validity was supported by the use of official guidelines and an expert panel of nephrologists to develop content and improvement in mean test performance with increasing level of training (PGY1 59.2 +/- 13.5%, PGY2 62.6 +/- 12.3% and PGY3 64.3 +/- 12.2%; P = 0.002). The reliability coefficient for the questionnaire instrument (Cronbach's alpha) was 0.69. CONCLUSION: Our brief questionnaire is a valid and reliable instrument to assess knowledge of CKD guidelines among internal medicine residents and identify specific gaps for improvement. PMID- 19674227 TI - To learn about a hospital's quality, one should look beyond its doors. PMID- 19674228 TI - In search of health. PMID- 19674229 TI - Seeking a new biomedical model. How evolutionary biology may contribute. AB - RATIONALE: The medical profession is not to blame for the limitations of the biomedical model with which it is often associated; the biology upon which that model is built is incomplete and bears some of the responsibility. Some of the more fundamental aspects of biological theory which are currently missing from the biomedical model need to be introduced in order to help provide a better description of the integrated biology involved. AIMS AND OBJECTIVES: By considering the biological nature of the individual human being as an organism, this article introduces a brief sketch of a new biomedical model of the individual as a biological entity. Core to this model is the idea of individual survival in the 'struggle for existence' introduced 150 years ago by Charles Darwin in 'On the Origin of Species'. PMID- 19674230 TI - Understanding health from a complex systems perspective. AB - Doctors often use theory to inform medical practice. The current bio-psycho social model of health may be advanced still further with theoretical rigour. Traditional fields of thermodynamics and newer fields of non-linear dynamics including chaos theory and complex systems science can inform our understanding of the complexity of human health, illness and disease. Commonly accepted aspects of human health may be projected as probabilities over time creating curves of human health potential. Maximum health may be represented by maximum functional complexity. Complexity's relationship to entropy and energy can produce a complex surface that better models the human experience of health and illness from birth to death. Such a potential health trajectory uniting complexity and entropy expands upon earlier theories of health while allowing for unusual predictions and the novel opportunity to test and validate this model of human health. PMID- 19674231 TI - Lessons from Jurassic Park: patients as complex adaptive systems. AB - RATIONALE: With realization that non-linearity is generally the rule rather than the exception in nature, viewing patients and families as complex adaptive systems may lead to a better understanding of health and illness. Doctors who successfully practise the 'art' of medicine may recognize non-linear principles at work without having the jargon needed to label them. METHODS: Complex adaptive systems are systems composed of multiple components that display complexity and adaptation to input. These systems consist of self-organized components, which display complex dynamics, ranging from simple periodicity to chaotic and random patterns showing trends over time. RESULTS: Understanding the non-linear dynamics of phenomena both internal and external to our patients can (1) improve our definition of 'health'; (2) improve our understanding of patients, disease and the systems in which they converge; (3) be applied to future monitoring systems; and (4) be used to possibly engineer change. CONCLUSION: Such a non-linear view of the world is quite congruent with the generalist perspective. PMID- 19674232 TI - What can complexity do for diabetes management? Linking theory to practice. AB - BACKGROUND: Diabetes presents a multifaceted picture with its rapidly rising prevalence associated with changing demographics and increasing levels of obesity in the developed world. Deaths from diabetes are predicted to rise by 25% over the next 10 years. The enormity of this public health challenge has been recognized the world over, but little attention has been paid to the theoretical frameworks underpinning practical management. AIM: This paper aims to introduce complexity theory and discuss its practical application to diabetes, focusing on a single 'tool' to provide an example of how theory can be linked to practice. APPLICATION: Critics have questioned the all inclusive nature of complexity seeing it as an intangible concept that fails to offer anything new to health care. However, few have appraised its practical application to a chronic disease that is currently managed using an outdated, linear, reduce and resolve model which fails to address the multiple interacting systems inherent within this condition. DISCUSSION: This article proposes that complexity theory provides an interprofessional perspective for describing and understanding the processes involved, and provides working 'tools' for patients, carers and practitioners that capture the reality of managing this chronic disease in modern life. PMID- 19674233 TI - The personal nature of health. PMID- 19674234 TI - 'It just doesn't seem to fit'. Environmental illness, corporeal chaos and the body as a complex system. AB - Environmental illness (EI) still remains something of an enigma, despite attempts to squeeze it into the increasingly flexible framework of the biomedical model or label it as 'psychogenic'. Consistently, environmentally ill bodies fail to respond to conventional medical tests, with the emergence of ambiguous, indecipherable or negative results. During a series of in-depth interviews in Canada, both patients and environmental health practitioners advocated the need for an alternative paradigm from which to view EI. They also spoke of the complex nature of the illness, a conundrum that is currently being explored at the Nova Scotia Environmental Health Centre through an alternative epistemological framework: complexity theory. In this paper, I present a tentative proposal, based on the juxtaposition of theoretical insights from this new science to discourses emanating from patients and doctors. Specifically, I argue that the diverse symptoms of EI can be comprehended through a study of complexity theory, a conception that has positive implications for treatment. PMID- 19674235 TI - Combined dextromethorphan and chlorpheniramine intoxication in impaired drivers. AB - Dextromethorphan is a nonprescription antitussive which has been gaining in popularity as an abused drug, because of the hallucinogenic, dissociative, and intoxicating effects it produces at high doses. This report describes a series of eight drivers arrested for driving under the influence of the combined effects of dextromethorphan and chlorpheniramine, and a further four drivers under the influence of dextromethorphan alone. In the combined dextromethorphan/chlorpheniramine cases, blood dextromethorphan concentrations ranged from 150 to 1220 ng/mL (n = 8; mean 676 ng/mL, median 670 ng/mL), and chlorpheniramine concentrations ranged from 70 to 270 ng/mL (n = 8; mean 200 ng/mL, median 180 ng/mL). The four cases without chlorpheniramine present had blood dextromethorphan concentrations between 190 and 1000 ng/mL (mean 570 ng/mL, median 545 ng/mL). Some drivers had therapeutic concentrations of other drugs present. Drivers generally displayed symptoms of central nervous system (CNS) depressant intoxication, and there was gross evidence of impairment in their driving, including weaving, leaving the lane of travel, failing to obey traffic signals, and involvement in collisions. Drug Recognition Expert opinions confirmed that the subjects were under the influence of a drug in the CNS depressant category. PMID- 19674236 TI - Sequences of the cytochrome C oxidase subunit I (COI) gene are suitable for species identification of Korean Calliphorinae flies of forensic importance (Diptera: Calliphoridae). AB - Calliphorinae fly species are important indicators of the postmortem interval especially during early spring and late fall in Korea. Although nucleotide sequences of various Calliphorinae fly species are available, there has been no research on the cytochrome c oxidase subunit I (COI) nucleotide sequences of Korean Calliphorinae flies. Here, we report the full-length sequences of the COI gene of four Calliphorinae fly species collected in Korea (five individuals of Calliphora vicina, five Calliphora lata, four Triceratopyga calliphoroides and three Aldrichina grahami). Each COI gene was amplified by polymerase chain reaction and directly sequenced and the resulting nucleotide sequences were aligned and analyzed by MEGA4 software. The results indicate that COI nucleotide sequences can be used to distinguish between these four species. Our phylogenetic result coincides with recent taxonomic views on the subfamily Calliphorinae in that the genera Aldrichina and Triceratopyga are nested within the genus Calliphora. PMID- 19674237 TI - Characteristic features of pediatric firearm fatalities--comparisons between Australia and the United States. AB - Gunshot deaths in children less than 17 years of age from Adelaide, SA, Australia (1969-2005) and from San Diego County, CA, United States (1988-2005) were compared. Forty-two pediatric gunshot fatalities occurred in South Australia (1.1 cases/year; M:F = 30:12). There were 6 accidents (14%), 14 suicides (33%), and 22 homicides (52%). In San Diego there were 185 cases (c.10 cases/year; M:F = 148:37). There were 6 accidents (3%), 42 suicides (23%), 130 homicides (70%), and 7 undetermined cases (4%). The incidence of homicide was significantly higher in San Diego County compared to Adelaide (p < 0.001), with a higher proportion of murder-suicides occurring in Adelaide. There were markedly more accidents and suicides involving males in Adelaide and a far higher number of male homicide victims in San Diego County compared to females. Rifles of 0.22-caliber were preferred weapons in South Australia, compared to handguns in San Diego County. PMID- 19674238 TI - Heroin overdose deaths and heroin purity between 1990 and 2000 in Istanbul, Turkey*. AB - Turkey has continuously experienced problems with abuse of, and addiction to, opium derivatives. In this study, we analyzed the relationship between heroin overdose deaths and the characteristics of seized opium derivatives. Data were gathered from the Council of Forensic Medicine of the Ministry of Justice in Istanbul from 1990 to 2000. There were 636 heroin-related deaths during this period, 595 of which were classified as heroin overdose deaths. Mean crude and weighted heroin purities remained relatively constant and were calculated to be 46% (57-34%) and 51% (39-59%), respectively. The weight of heroin and the number of heroin seizures, but not the heroin purity, were significantly associated with the number of heroin-related deaths. Prevention strategies are needed to reduce the number of deaths caused by overdoses in countries situated on drug trafficking routes. These strategies should focus on drug trafficking, by providing increased levels of, and support for, law enforcement, stopping the supply of precursor chemicals, and combating corruption among border officials. PMID- 19674239 TI - Morphine concentrations in stomach contents of intravenous opioid overdose deaths. AB - Death caused by heroin overdose is almost always the result of intravenous injection of the drug in Australia. We briefly describe a case where a heroin overdose was initially thought to be the result of oral ingestion of the drug, primarily as a result of higher concentrations of morphine in stomach contents than in blood. During the subsequent criminal trial and investigation, however, the issue of the entero-hepatic circulation of morphine was raised as a possible reason for the presence of morphine in the stomach contents. In this study, we report on the distribution of opioids in blood, stomach contents, urine, liver, and bile in 29 deaths caused by intravenous heroin overdose. The mean total and free blood morphine concentrations were 0.60 and 0.32 mg/L, respectively, and the mean stomach contents total morphine concentration was 1.16 mg/kg. All cases had detectable morphine in the stomach contents, and 24 of 29 cases (83%) had higher concentrations of total morphine in stomach contents than in blood. The mean total morphine concentration in bile was c. 100 times that in blood, and the liver total morphine concentration averaged twice that of blood levels. We conclude that the entero-hepatic circulation of morphine and subsequent reflux of duodenal contents back into the stomach can result in the deposition of morphine in gastric contents. Consequently, the relative levels of opioids in blood and stomach contents cannot be used to determine the site of administration of the drug. PMID- 19674240 TI - Preventing suicide in jails and prisons: suggestions from experience with psychiatric inpatients. AB - Both among psychiatric inpatients and inmates of prisons and jails, suicide is highly prevalent with alarming rates. In many countries, there has been a call for action to prevent such deaths and to educate staff in the early recognition of suicide risk. A careful MedLine search was used to identify relevant papers dealing with suicide prevention in psychiatric inpatients. This paper reviews this research and the policy recommendations that have been developed for psychiatric hospitals in order to reduce the incidence of suicide in their patients. Results derived from this search indicated that these policy recommendations can be applied to suicide prevention in correctional settings, and it is argued that suicide prevention programs in correctional settings can benefit from the research conducted and the policy recommendations for suicide prevention in psychiatric facilities. In conclusion, the best practices for preventing suicides in jail and prison settings should include the following elements: training programs, screening procedures, communication between staff, documentation, internal resources, and debriefing after a suicide. PMID- 19674241 TI - Pseudo-stab wounds: putrefactive dehiscence of remote surgical incisions masquerading as stab wounds. AB - Artifacts due to decomposition can be mistaken for antemortem injury leading to an incorrect suspicion of foul play. We describe an instance of postmortem wound dehiscence that mimicked antemortem stab wounds. A man with a history of colon cancer and substance abuse was found dead. There were advanced putrefactive changes and multiple defects of the anterior torso that resembled stab wounds. Subsequent investigation revealed that 11 months earlier, he had a laparoscopic assisted colon resection that involved surgical incisions corresponding in location and size to the above defects. Putrefactive gases may cause bloating of the body. This distension may cause recent and remote healed incisions to dehisce. Correlation of these "defects" at autopsy with the antemortem clinical history is crucial in determining their etiology. PMID- 19674242 TI - Foreigners dying in Istanbul. AB - The study included 411 deaths selected from 14,647 medicolegal deaths autopsied in the Morgue Department of Forensic Medicine Institute Directorate, affiliated with the Ministry of Justice, between 1998 and 2002. Data were collected from court documents, coroner's investigation reports, and autopsy reports. The parameters of age, gender, nationality and origin, cause and place of death in foreigners dying in Istanbul were evaluated in the study. Out of 14,647 medicolegal deaths, 3.5% were foreigners from 34 different nationalities. The nationality with the highest rate of foreigner deaths (34%) was Romanian. Out of 411 deaths, 74.3% were male and 25.7% were female. Of all cases, 64.4% were tourists visiting Istanbul and 35.6% had a job in Istanbul. Of 146 foreigners employed in Istanbul, 94.5% did not have a work permit, while only 5.5% had a work permit. PMID- 19674243 TI - High prevalence of obesity in ambulatory children and adolescents with intellectual disability. AB - BACKGROUND: Obesity prevalence is unusually high among adults with intellectual disability (ID). There is limited and conflicting evidence on obesity prevalence among ambulatory children and adolescents with ID. The present study aimed to estimate obesity prevalence in this group and to compare with population prevalence. METHODS: Survey of nine schools (n = 206, 150 boys) for ambulatory children and adolescents with mild-moderate ID in Scotland in 2007. Obesity was defined as measured body mass index (BMI) at or above the 95th percentile relative to UK 1990 reference data, and using the international definition based on BMI. Obesity prevalence observed was compared against Scottish population data on obesity prevalence from the most recent nationally representative survey. RESULTS: Obesity prevalence (at or above 95th percentile for BMI) was 36%, and was significantly higher among those attending secondary schools compared with primary schools (P < 0.01). Prevalence of obesity was significantly higher than in the general paediatric population in both boys and girls (P < 0.01). CONCLUSIONS: The present study suggests that that obesity may be very prevalent among ambulatory children and adolescents with ID, and that increased obesity risk may begin in childhood. PMID- 19674244 TI - Relation between door-to-balloon time and microvascular perfusion as evaluated by myocardial blush grade, corrected TIMI frame count, and ST-segment resolution in treatment of acute myocardial infarction. AB - BACKGROUND: The role of microvascular perfusion in relation to door-to-balloon time and mortality remains unknown. We sought to compare microvascular perfusion in patients who had undergone primary percutaneous coronary intervention with door-to-balloon time < or =90 minute versus >90 minutes. METHODS: Using myocardial blush grade (MBG), corrected TIMI frame count (CTFC), and ST-segment resolution, microvascular perfusion was assessed in 297 patients who underwent successful primary percutaneous coronary intervention between January 2007 and April 2008. RESULTS: Door-to-balloon time was < or =90 minutes in 199 (67%) patients and >90 minutes in 98 (33%) patients. Univariate analysis showed that door-to-treatment >90 minutes was associated with MBG 0/1 and CTFC >28. However, it was not associated with ST-segment resolution < or =70%. After adjustment for baseline confounding factors, door-to-balloon time >90 minutes was still associated with MBG 0/1 (adjusted OR 3.20, 95% CI 1.87 to 5.49, P < 0.001) and CTFC >28 (adjusted OR 6.30, 95% CI 3.56 to 11.17, P < 0.001). Thirty-day mortality was higher in patients with longer door-to-balloon time (adjusted OR 2.87, 95% CI 0.94 to 8.77, P = 0.064). CONCLUSION: We found that door-to-balloon time >90 minutes, compared with < or =90 minutes, was independently associated with MBG 01/2 and CTFC >28, both suggesting microvascular obstruction. However, such association was not found in ST-segment resolution. Patients with door-to balloon time >90 minutes also had higher 30-day mortality. PMID- 19674245 TI - Unusual presentation of sarcoidosis: solitary intracranial mass lesion mimicking a glioma. AB - We present a patient with sarcoidosis with an isolated intraparenchymal mass lesion that was similar to a glioma on magnetic resonance imaging. On fluid attenuated inversion recovery images, a small hyperintense signal change in the right uncus was observed. Three months later, enlargement of the abnormal signal lesion was observed. An initial diagnosis of glioma was made. A biopsy of the temporal lobe tumor was done. On histology, a noncaseating granulomatous inflammation consistent with neurosarcoidosis was diagnosed. Albeit rarely, we should consider the possibility of neurosarcoidosis in the differential diagnosis of isolated intraparenchymal mass lesion, when the mass is located beside the pia mater. PMID- 19674246 TI - Evaluation of the effects of thyrotropin releasing hormone (TRH) therapy on regional cerebral blood flow in the cerebellar variant of multiple system atrophy using 3DSRT. AB - BACKGROUND: Thyrotropin releasing hormone (TRH) improves cerebellar ataxia and cerebellar perfusion in patients with spinocerebellar degeneration. It is not known whether TRH therapy can improve the cerebellar regional cerebral blood flow (rCBF) or not in patients with cerebellar variant of multiple-system atrophy (MSA C). PATIENTS AND METHODS: Seven patients with MSA-C received TRH intravenously (2 mg/day) for 14 days. Clinical efficacy was assessed using the International Cooperative Ataxia Rating Scale (ICARS) and brain perfusion single photon emission-computed tomography was performed before and after therapy. The rCBF in each region of interest (ROI) was calculated using 3DSRT, a fully automated the ROI technique. RESULTS: The ICARS scores slightly improved in 6 of the 7 patients after TRH therapy, but this was not statistically significant. After TRH therapy, the cerebellar rCBF reduced in the 6 of 7 patients and the mean rCBF in cerebellum also significantly decreased (P=0.029, paired t-test), whereas the rCBF in the precentral segment tend to increase (P=0.048, paired t-test). CONCLUSION: TRH therapy may be less effective on cerebellar ataxia and cerebellar rCBF in MSA-C. The 3DSRT program may be useful for the evaluation of the efficacy of TRH therapy on cerebral blood flow. PMID- 19674247 TI - DWI lesion volume reduction following acute stroke treatment with transient partial aortic obstruction. AB - BACKGROUND AND PURPOSE: Diffusion-weighted imaging (DWI) identifies acute cerebral ischemia and DWI lesions are thought to indicate irreversibly damaged areas. However, new evidence suggests that DWI lesions may be reversible, especially with reperfusion. We present a patient who showed substantial reversal of her acute DWI lesion following partial aortic occlusion with NeurofloTM, a novel dual balloon catheter (NeurofloTM, CoAxia, MN). METHODS: Case report/literature review. RESULTS: A 48-year-old woman presented with left-sided weakness and demonstrated an acute DWI lesion in the right middle cerebral artery territory, with diffusion-perfusion mismatch. She was enrolled into an experimental study in which a dual balloon catheter was inflated in the lower aorta. The patient improved and her postprocedure magnetic resonance image showed a significant reduction in lesion volume on diffusion and perfusion-weighted imaging. At 1 month, a repeat computed tomography scan showed a small infarction in the right insula, lentiform nucleus, and frontal cortex. The patient had recovered with no significant disability at her 3-month follow-up. CONCLUSION: Reperfusion can improve DWI lesions. Partial aortic obstruction with a novel dual balloon catheter may be useful to promote reperfusion. PMID- 19674248 TI - Multifocal intradural spinal AVF and renal artery aneurysms in a case of Klippel Trenaunay Syndrome (KTS). AB - We report an interesting case of a young patient who had hypertrophy of right leg and nevoid geographic skin lesion on the dorsal aspect of the right foot and leg suggestive of Klippel Trenaunay syndrome (KTS) and who presented for spinal digital subtraction angiography (DSA) to investigate the cause of progressive weakness of bilateral lower limbs. DSA revealed spinal arteriovenous fistulae (AVFs) at 3 levels and bilateral renal artery aneurysms. Although multiple intradural spinal cord AVFs and renal artery aneurysms are considered a feature of KTS, their clear demonstration in a single case either alone or together is not available in literature to the best of this author's knowledge. Existence of multifocal intradural fistulae in KTS calls for a detailed and complete spinal DSA even after a fistula is detected. PMID- 19674249 TI - Morphometric analysis of CD34-positive vessels in salivary gland adenoid cystic and mucoepidermoid carcinomas. AB - BACKGROUND: Carcinomas of the salivary glands are uncommon and morphologically a diverse group of malignancies. To evaluate the prognostic value of CD34 immunostaining of the vessels in adenoid cystic carcinoma (AdCC) and mucoepidermoid carcinoma (MEC), an automated image analysis method was used. METHOD: In a nationwide study, covering salivary gland cancer (SGC) patients in Finland 1991-1996, 37 AdCC and 18 MEC patients (M 25, F 30, age 25-90, mean 63) were included. In addition to clinical characteristics the size, shape, staining intensity and vessel density in CD34 immunostained histologic samples were measured. RESULTS: Altogether 4433 vessels were measured from AdCC and 2615 from MEC tumor. Of the total tumor vessels measured, 2651 were from patients who deceased with disease (Group I) and 4397 were from specimens derived from those who did not die of disease (Group II) during the 10-year follow-up. The staining intensity was significantly higher in MEC than in AdCC tumor (P = 0.0005). In MEC, the Group I patients had a higher staining intensity among high-grade patients compared with patients with low grade disease, whereas the tumors in Group II had a lower staining intensity among the high-grade compared with the low grade tumors (P = 0.018). A higher vessel density was found in patients with MEC in group II compared with group I (P = 0.017). CONCLUSIONS: The staining intensity of CD34 positive vessels in MEC was higher than in AdCC. In MEC, higher staining intensity of vessels in high-grade tumors and lower vessel density in all MEC patients, predicted poor survival. PMID- 19674250 TI - Erectile dysfunction (ED) is a shared sexual concern of couples II: association of female partner characteristics with male partner ED treatment seeking and phosphodiesterase type 5 inhibitor utilization. AB - INTRODUCTION: Erectile dysfunction (ED) is a prevalent condition that impacts on both patients and their female partners. ED may therefore be regarded as a shared sexual concern for couples. The current analysis of the Female Experience of Men's Attitudes to Life Events and Sexuality (FEMALES) study data addresses women's perceptions, beliefs, and attitudes concerning their partner's ED, and whether these are associated with the likelihood of the male partner seeking medical advice and utilizing phosphodiesterase type 5 inhibitors. AIM: The current research sought to explore the association of female partners' perceptions of male partners' ED and male partners' medical consultation and treatment seeking for ED. METHODS: Questionnaires were sent to partners of men who participated in the Men's Attitudes to Life Events and Sexuality (MALES) 2004 study, and who consented to their partner's involvement. A modified version of the questionnaire used in the MALES study was developed for the FEMALES study, reflecting the female partner's perspective. MAIN OUTCOME MEASURE: A 65-item questionnaire assessing women's perceptions, beliefs, and attitudes regarding various aspects of ED. RESULTS: Women's perceptions of the nature and causes of their partner's ED were significantly associated with men's treatment seeking and utilization. Significant associations were observed between women's level of satisfaction with the relationship before ED onset; perceptions of the impact of ED on quality of life; desire to deal with ED; attitudes to ED treatment; and the treatment-seeking behavior of the male partner. Multivariate regression analyses identified a mixture of female and male partner perceptions and attitudes that uniquely accounted for >30% of the variance in men's ED treatment-seeking behavior and treatment utilization. CONCLUSION: This study illustrates the importance of the female partner's attitudes to ED in men's ED treatment-seeking behavior. These findings strongly support the potential benefits of partner integration into ED consultation and treatment strategies. PMID- 19674251 TI - Erectile dysfunction: initial symptom of a patient with lung cancer. AB - INTRODUCTION: Penile metastases are rare and represent the advanced stage of the primary tumor. The patients usually have a history of a previously diagnosed malignancy and when metastasis to penis occurs, the most common findings would be priapism, pain, and difficulty in voiding. AIM: We aimed to present a patient who had erectile dysfunction as the initial symptom of lung cancer. Besides the unusual clinical presentation, the sonographic and magnetic resonance imaging (MRI) findings of the penile metastasis were also not typical. METHODS: A 57-year old man with erectile dysfunction was admitted to the Department of Urology. On physical examination, there was a rigid, smooth, immobile, and painless mass at the base of the corpora cavernosa. Ultrasonography and MRI were performed in order to delineate the nature of the lesion. RESULTS: Radiological findings could not lead to a certain diagnosis and the lesion could not be resected completely during the surgery. Therefore, biopsy of the corpus cavernosum penis was performed. The histopathological diagnosis was metastatic malignant epithelial tumor consistent with nonsmall cell carcinoma. Further investigations revealed a metastatic lung cancer. CONCLUSIONS: Penile metastasis may rarely be the initial presentation of a malignancy and erectile dysfunction may be a seldom symptom. PMID- 19674252 TI - Endothelial antioxidant administration ameliorates the erectile response to PDE5 regardless of the extension of the atherosclerotic process. AB - INTRODUCTION: The lack of phosphodiesterase type 5 inhibitor effects in patients with erectile dysfunction (ED) of arterial origin may be caused by an endothelial dysfunction that causes a series of biochemical alterations leading to a reduced nitric oxide (NO) bioavailability and increased oxidative stress. AIM: The aim of this study was to evaluate the effects of the treatment with endothelial antioxidant compounds (EAC) on the erectile response to sildenafil in patients with arterial ED already treated with sildenafil (100 mg twice a week for 8 weeks). MEAN OUTCOME MEASURES: A patient was considered responsive when the 5 item International Index of Erectile Function questionnaire score increased by >5 points. METHODS: Fifty-three patients with arterial ED, hypertension, and diabetes mellitus were randomly given, for 8 weeks, EAC (1 dose/day) and, after a wash out of 8 weeks, sildenafil (100 mg) plus EAC. The patients were divided into the following four groups: A (N = 12): patients with ED alone; B (N = 14): patients with ED plus atheromasic plaques and/or increased intima-media thickness of common carotid arteries; C (N = 14): patients with ED plus lower limb artery abnormalities; and D (N = 13): patients with ED plus carotid and lower limb artery abnormalities. RESULTS: The administration of EAC plus sildenafil resulted in a significantly higher number of responsive patients (N = 36, 68%) compared with sildenafil alone (N = 24, 45%) or EAC alone (N = 17, 32%). The percentage of patients who successfully responded to the combined treatment increased in the various groups. It was 83%, 64%, 71%, and 54%, respectively, for groups A, B, C, and D. Furthermore, patients treated with EAC and sildenafil reached a successful response in a shorter length of time (3 weeks) compared with patients responsive to sildenafil (5.2 weeks) or EAC (5.7 weeks) alone. CONCLUSION: EAC administration to patients with arterial ED improved the success rate to sildenafil. These data suggest that, in such patients, a combined treatment may be considered to increase bioavailable NO and to neutralize radical oxygen species, which in turn inactive NO. PMID- 19674253 TI - Persistent genital arousal disorder and trazodone. Morphometric and vascular modifications of the clitoris. A case report. AB - INTRODUCTION: Persistent genital arousal disorder (PGAD) is an unwanted genital arousal which occurs in absence of sexual interest and desire. AIM: To report a case of PGAD presumably due to the use of trazodone in a young eumenorrheic woman. METHODS: A young (29 years old), eumenorrheic (menstrual cycle of >25 and <35 days) woman suffered of unwanted genital arousal and uncontrollable orgasms. In the past, the patient undertook trazodone treatment. The patient was submitted, in the periovulatory (day 12) phase of the menstrual cycle, to bi- and tri-dimensional ultrasonographic and color Doppler analyses of the clitoral structures prior and after an unwanted orgasm. MAIN OUTCOMES MEASURES: 2D ultrasonographic evaluation of the clitoral body volume and color Doppler evaluation of the dorsal clitoral arteries; 3D power Doppler reconstruction of the clitoral vascularization. RESULTS: The clitoral volume was 1.33 mL before the orgasm and resulted 1.36 mL and 1.33 mL, respectively after 1 minute and 15 minutes from the orgasm. The Pulsatility Index (PI) of the dorsal clitoral artery was 1.05 before the orgasm. It resulted lower after 1 minute (PI = 0.82) and 15 minutes (PI = 0.85) from the orgasm. CONCLUSIONS: A subtle and intermittent clitoral priapism may favor the feeling of arousal persistence and elicit unbidden and unwelcomed orgasms. PMID- 19674254 TI - The association between varicocele, premature ejaculation and prostatitis symptoms: possible mechanisms. AB - INTRODUCTION: No study has ever systematically evaluated the impact of varicocele on sexual function. AIM: Two cross-sectional studies were performed in patients attending an andrology unit either for male sexual dysfunction (study 1) or couple infertility (study 2). In study 1, we evaluated the impact of varicocele on sexual function. In study 2, we retrospectively evaluated a possible association between varicocele and prostatitis signs and symptoms. METHODS: Study 1 refers to a consecutive series of 2,448 (mean age 52.0 +/- 12.9 years) subjects. Study 2 consists of a consecutive series of 139 male subjects (mean age 37.3 +/- 6.3). MAIN OUTCOME MEASURES: In study 1, varicocele was clinically classified into three grades according to Dubin criteria. Different hormonal parameters were also evaluated. All the patients of study 2 underwent simultaneous scrotal and transrectal color-Doppler ultrasonography (CDU) along with seminal characteristics and interleukin-8, a surrogate marker of prostatitis. RESULTS: After adjusting for age, subjects with severe varicocele (N = 284, 11.6%; Dubin grade 2 and 3) showed a reduction of testicular volume (P < 0.01), higher luteinizing hormone (LH) (P < 0.05), follicle stimulating hormone (FSH) (P < 0.0001) and prolactin (P < 0.05) levels, and also an enlarged or tender prostate at digito-rectal examination (P < 0.05). Premature ejaculation was the only sexual symptoms significantly associated with varicocele (29.2% vs. 24.9% in subjects with or without varicocele, respectively; P < 0.05). In study 2, subjects with severe echographic-defined varicocele (basal venous reflux increasing or not after Valsalva's maneuver; N = 28, 20.1%) showed CDU features of prostatitis and higher seminal inteleukin-8 levels. The presence of any degree of varicocele (N = 40, 28.8%) was also associated with prostatitis symptoms, as measured by the National Institutes of Health Chronic Prostatitis Symptom Index scoring (P < 0.05), and in particular with the pain domain (P < 0.05). CONCLUSIONS: In conclusion, signs and symptoms of prostatitis are more common in varicocele patients, who more often complain of premature ejaculation. PMID- 19674255 TI - Erectile dysfunction in heart failure: correlation with severity, exercise performance, comorbidities, and heart failure treatment. AB - INTRODUCTION: Erectile dysfunction (ED) is frequent in males with chronic heart failure (HF) with a severe impact on quality of life for many individuals. The correlation of ED with age and HF severity, comorbidity, and treatment is unclear. AIM: We evaluated the correlation between ED and HF severity, treatment, and comorbidity. METHODS: One hundred one HF patients aged < or =70 years, with left ventricular ejection fraction < or =40%, and stable clinical condition took part in the study. We measured: (i) hemoglobin, glycemia, glicated hemoglobin, creatinine, cholesterol, thyroid-stimulating-hormone, C-reactive-protein, total/free testosterone; (ii) ED, depression, urological symptoms, and signs of low testosterone by means of questionnaires; and (iii) HF severity by means of echo, brain natriuretic peptide, and cardiopulmonary exercise test. MAIN OUTCOME MEASURES: ED was measured by means of International Index of Erectile Function-5 questionnaire and its score was correlated with exercise cardiopulmonary test parameters, HF severity, treatment and HF comorbidities. RESULTS: ED prevalence was 69.3%, 81.1%, and 56% in total population and in patients with and without coronary artery lesions, respectively. ED was absent in 31 while it scored mild, mild to moderate, moderate and severe in 15, 18, 12, 25 individuals, respectively. Sexual activity requires, in the orgasmic phase, an oxygen consumption (VO(2)) between 10 and 14 mL/min/kg. In none of the individuals with peak VO(2) < 10 mL/min/kg was sexual function normal or slightly impaired, while in 10/29 of patients with peak VO(2) between 10 and 14 mL/min/kg there was a normal or slightly reduced sexual performance. On monovariable analysis, several parameters were correlated with ED, but at multivariable analysis only age (P = 0.002), hemoglobin (P = 0.042), diabetes (P = 0.040), and use of diuretics (P = 0.052) remained so. CONCLUSIONS: ED is frequent in HF. A normal or only slightly impaired sexual activity is possible with peak VO(2) > 10 mL/min/kg. On multivariable analysis, only age, diabetes, use of diuretics, and hemoglobin are related to ED. PMID- 19674256 TI - Timing of dose relative to sexual intercourse attempt in previous sildenafil citrate users treated with tadalafil: a geographical comparison from a single arm, open-label study. AB - INTRODUCTION: Previous research has demonstrated that sildenafil citrate users alter dosing-sexual attempt behavior when switched to tadalafil. The impact of geography and culture on sexual behavior with phosphodiesterase type 5 (PDE5) inhibitor treatment has not been fully investigated. AIM: To describe and compare the changes in dosing-sexual attempt behavior with sildenafil citrate vs. tadalafil treatment across four distinct geographies: Asia, Australia/New Zealand (ANZ), Central Eastern Europe/Middle East (CEE/ME), and Latin America (LA). METHODS: Data from a single-arm, open-label clinical trial conducted in 21 countries from November 2002 to May 2004 were used in this analysis. Men with erectile dysfunction and a history of > or =6-week prior sildenafil citrate use continued sildenafil citrate treatment for 4 weeks then switched to tadalafil for 8 weeks. Dosing instructions were provided. MAIN OUTCOMES MEASURES: Timing of dose and sexual intercourse was assessed through patient diaries for the final 4 weeks of each treatment period. RESULTS: A total of 2,760 men were enrolled: Asia 15.8%; ANZ 29.4%; CEE/ME 19.7%; LA 35.1%. The median time from dosing to intercourse was significantly increased during tadalafil treatment across all geographical regions; however, the magnitude of increase differed significantly by geography (P < 0.0001). The Asian cohort demonstrated the shortest duration between dosing and sexual intercourse attempts (irrespective of drug), and altered sexual behavior the least upon switching to tadalafil. The ANZ cohort demonstrated the longest duration between dosing and sexual intercourse attempts (irrespective of drug), and altered sexual behavior the most upon switching to tadalafil. CONCLUSION: Men with a history of established sildenafil citrate use alter their dose-attempt behavior when treated with tadalafil irrespective of geography. However, the extent to which sexual behavior alters is not uniform across geographical regions, suggesting that dosing instructions and duration of drug effectiveness, in combination with personal and cultural preferences, may determine sexual behavior with PDE5 inhibitor use. PMID- 19674257 TI - Nitric oxide modulates the discharge rate of basal forebrain neurones: a study in freely moving rats. AB - In urethane-anaesthetized rats the infusion of a nitric oxide (NO)-donor [NOC-18, 1 mM (DETA/NO); 2,2'-(hydroxynitrosohydrazino)bis-ethanamine)] into the basal forebrain (BF) inhibited the discharge rate of most neurones, suggesting that NO may promote sleep via inhibition of wake-promoting neurones in the BF. However, this hypothesis still needs to be confirmed in freely moving rats. The objective of this study was to examine whether NO modulates the discharge rate of BF neurones in freely moving rats in a similar manner to anaesthetized rats. We measured the discharge rates of BF neurones in freely moving rats during microdialysis infusion of a NO-donor (1 mm; NOC-18) in different vigilance states. Neurones were characterized as wake (W)-on (51.8%), W-off (28.6%) and W/non-rapid eye movement (REM)-independent (21.4%) based on their discharge profiles during wakefulness (W) and non-REM sleep. The NO-donor affected the discharge rate of most BF neurones during quiet wakefulness (QW; 55%) and non-REM sleep (64%). The most prominent response in all neuronal groups was a decrease in the discharge rate during QW and non-REM sleep. A small subpopulation of neurones increased the discharge rate. The increase in NO in the BF during prolonged wakefulness may facilitate sleep via inhibition of wake-promoting neurones. PMID- 19674260 TI - Minimizing pathogen transmission at primate ecotourism destinations: the need for input from travel medicine. PMID- 19674261 TI - Hepatitis A risk in travelers. AB - BACKGROUND: Traveling to highly endemic areas for hepatitis A is increasing while the immunization level in travelers has been shown to be low in the countries studied. METHODS: In this population-based study, we have estimated the incidence rate of travel-related hepatitis A during 1997 to 2005 by use of the Swedish notification system of communicable diseases and an ongoing national database on travel patterns. We have also acquired airport-based immunization data from 2007. RESULTS: During the study period, 636 cases of travel-related hepatitis A were notified. Traveling to East Africa was associated with the highest incidence rate (14.1 cases/100,000 person months), followed by the Middle East (5.8/100,000 person months), and India with neighboring countries (5.6/100,000 person months). Visiting Friends and Relatives (VFR) travelers represented 83, 91, and 70% of the cases to these three regions. By age-group, the highest incidence was found in children 0 to 14 years (3.1/100,000 travelers) where 88% of the cases were VFR travelers. Incidence rate in unprotected travelers to East Asia, North Africa, and the Middle East was 2, 12, and 18 cases/100,000 person months, respectively. In 2007, 79% of the travelers were immunized against hepatitis A. CONCLUSIONS: We conclude that travelers, and especially children, who are VFR in endemic areas constitute a high-risk group for acquiring hepatitis A infection, while the risk for unprotected tourists to East Asia is low. PMID- 19674262 TI - Viral etiology of acute respiratory infections among Iranian Hajj pilgrims, 2006. AB - BACKGROUND: Every year more than 2 million pilgrims from different countries in the world including Iran participate in the annual Hajj in Saudi Arabia. Respiratory diseases have been the most common cause of illnesses among Iranian pilgrims. METHODS: Direct fluorescent staining and viral culture were performed on nasal wash specimens of Iranian Hajj pilgrims with symptoms of acute respiratory tract infections at Shiraz (a city in southern Iran) airport on return from the Hajj during December 2006 to January 2007. They were screened for influenza A and B, parainfluenza 1 to 3, adenovirus, and respiratory syncytial virus (RSV) by viral culture and immunofluorecent staining. Rhinovirus and enterovirus were diagnosed based on reverse transcription polymerase chain reaction methods. RESULTS: The patients aged between 19 and 82 years (mean: 52.4 years) consisting of 135 females and 120 males. Cough in 213(83.5%) and sore throat in 209 (82%) were the most common symptoms. Eighty-three patients (32.5%) had viral pathogens: influenza in 25 (9.8%), parainfluenza in 19 (7.4%), rhinovirus in 15 (5.9%), adenovirus in14 (5.4%), enterovirus in 5 (2%), and RSV in 4 (1.6%) and coinfection with two viruses in 1 patient (0.4%). Influenza virus was identified more in unvaccinated than in vaccinated pilgrims (16.5% vs. 9.2%) but statistically insignificant (p= 0.19). CONCLUSIONS: According to the results, each of the above-mentioned viruses played a role in the development of respiratory diseases among Iranian pilgrims, with influenza virus as the commonest one. Because influenza vaccine could not prevent respiratory infections in Hajj pilgrims statistically, the possibility of the appearance of new drift variants not included in vaccine and also inappropriate vaccine handling and storage might be considered. So it is also advisable to check if the circulating influenza strains were different from the vaccine strains. PMID- 19674263 TI - Overseas visitor deaths in Australia, 2001 to 2003. AB - BACKGROUND: The health and safety of international visitors remain an important issue for Australia and other tourist destinations. The death of visitors remains an important indicator of safety. The aim of this study was to provide updated figures on deaths of overseas travelers in Australia. METHODS: Data were sourced from the Australian Bureau of Statistics concerning deaths of overseas visitors for the years 2001 to 2003. RESULTS: There were 1,068 overseas visitor deaths (701 males, 66%) during the study period 2001 to 2003. Death by natural causes increased with age, while deaths associated with accidents were more frequent among younger age groups. The majority of deaths were from natural causes (782, 73%), particularly ischemic heart diseases (26%). There were a total of 247 accidental deaths (23% of all deaths) with the main causes being transportation accidents (14% of all deaths) and accidental drowning/submersion (5% of all deaths). The countries contributing the most deaths were the UK (247, 23%), New Zealand (108, 10%) Melanesia/Micronesia (95, 9%), and the United States (57, 5%). CONCLUSIONS: Australia remains a relatively safe destination for international travelers, at least in terms of fatalities, which appear to be declining. Most deaths of overseas tourists in Australia are due to natural causes with cardiovascular disease being the predominant cause of death in this group. Accidents remain the most common preventable cause of death of travelers, with road and water safety being the major issues. It is important that tourism and travel medicine groups continue to advocate for improved health and safety of international travelers visiting Australia. PMID- 19674264 TI - Clinicoepidemiological characteristics of HIV-infected immigrants attended at a tropical medicine referral unit. AB - BACKGROUND: Migration is a growing phenomenon with a well-known impact in infectious diseases epidemiology. Currently, immigrants represent almost 10% of the Spanish population. The majority come from countries where the prevalence of chronic viral illnesses is higher than in Spain. METHODS: To describe clinicoepidemiological features of human immunodeficiency virus (HIV)-infected immigrants attending our Unit and to compare differential characteristics depending on geographical origin, information from all new immigrants from January 1997 to December 2006 was collected. STUDY DESIGN: noninterventional retrospective chart review. RESULTS: We screened 1,609 patients of whom 77 (4.8%) were HIV antibody (Ab) positive. Of these, 80% were sub-Saharan Africans (SSAFR) and 20% were South-Central Americans (SCA). HIV prevalence was higher in SSAFR (5.6% vs 3.2%; p= 0.04). Overall, of those who were HIV Ab positive, 70% were male (median age 30 years), 59% heterosexuals, 9% hepatitis C virus coinfected, 8.6% hepatitis B virus coinfected, and 34% showed a positive tuberculin skin test. Median CD4 cell count was 263 cells/microL, median HIV-ribonucleic acid viral load 4.6 Log/mL, and 48% had a late diagnosis [acquired immunodeficiency syndrome (AIDS)-defining illness or <200 CD4 microL at the time of diagnosis]. Only 68% of patients for whom antiretroviral therapy was indicated actually started therapy and 22% were lost to follow-up just after diagnosis. SCA had lower CD4 cell counts (26 vs 168 cells/microL; p= 0.016), higher viral loads (5.3 vs 4.8 Log; p= 0.001), and were more likely to have an AIDS-defining illness (53% vs 21%; p= 0.04) compared to SSAFR. Tuberculin skin test reactivity was more common among SSAFR versus SCA [adjusted by CD4 count, odds ratio (OR) 6.3 and 95% confidence interval (CI): 0.65-60.5]. The main risk factor for late diagnosis was geographical origin: OR 4.6 (95% CI: 1.11-19.3) (SCA vs SSAFR; adjusted by the interval between the date of arrival in Spain and the date of HIV diagnosis). CONCLUSIONS: Almost half the HIV-infected immigrants were diagnosed in late stages. Patients were frequently lost to follow-up, and a significant minority did not start highly active antiretroviral therapy when indicated. SCA seem to have more severe immunosuppression at the time of diagnosis than SSAFR. Early voluntary routine HIV screening should be promoted. PMID- 19674265 TI - Sickle cell children traveling abroad: primary risk is infection. AB - BACKGROUND: Pediatricians taking care of sickle cell children in France are concerned about giving travel advice. Very few articles are published and no study has been done about it. A lot of pediatricians are using their own experience to decide if sickle cell children can travel abroad. Studying the consequences of such travel for sickle cell children is important to discuss common recommendations. METHODS: We conducted a prospective study from June 2006 to December 2007 on desires to travel expressed during our consultations with sickle cell children. We studied notable events that occurred during travel and at least 2 months after return. RESULTS: Of 52 desires to travel, 10 were cancelled. All of the 42 trips were to Africa. Median duration of travel was 1.29 months (0.5-3). Median age at travel was 7.6 years (0.2-17.7). Events during travel were two hospitalizations (4.8%), a transfusion (2.4%), and four paramedical or medical examinations (9.6%). After return, four events occurred: two SS children had Plasmodium falciparum malaria (4.8%) and two had digestive bacteremia (4.8%) in SC and Sbeta+ children. No event occurred during plane travel. None of our patients died. CONCLUSIONS: The primary risk for sickle cell children traveling to Africa is infection: malaria first and digestive septicemia second. These risks are increased by long travel and poor sanitary conditions. Each travel should be prepared a long time before departure, and each pediatrician should insist on malaria prophylaxis and sanitary conditions, especially for young children. Trips should be shorter than 1 month when possible. A longer prospective study will be done to confirm these results. PMID- 19674266 TI - Imported histoplasmosis: two distinct profiles in travelers and immigrants. AB - BACKGROUND: Histoplasmosis is acquiring importance in nonendemic areas due to the increase in travel and immigration, being the most common systemic mycosis acquired by European travelers. Epidemiological studies show that the incidence of Histoplasma infection in these patients may be higher than previously believed and a wide clinical spectrum of disease may be observed. METHODS: Cases of histoplasmosis diagnosed at a Tropical Medicine Referral Unit in Madrid, Spain, during the period January 1996 to December 2006 were reviewed. RESULTS: Ten cases of histoplasmosis in travelers and immigrants are described. Five HIV-positive patients (four immigrants and one expatriate) all presented with progressive disseminated disease. Five HIV-negative patients (travelers) all presented with pulmonary disease: four with an acute pulmonary form (one with pleural involvement) and one patient was found to have residual pulmonary disease (lung nodule). Three of the travelers also had rheumatologic manifestations (arthromyalgias or arthritis). CONCLUSIONS: Clinicians in nonendemic areas may be faced with patients with a diagnosis of histoplasmosis and although Histoplasma infection can have a varied and nonspecific clinical presentation, imported histoplasmosis may have two distinct profiles. Previously, healthy travelers may be exposed in endemic areas and mainly develop acute forms of the disease with a favorable outcome. Immigrants or expatriates from endemic areas who may be immunosuppressed due to HIV infection may experience reactivation of latent disease developing disseminated forms with high mortality rates. This infection should be considered in the differential diagnosis of diseases affecting travelers and immigrants. PMID- 19674267 TI - Does registration of professionals improve the quality of travelers' health advice? AB - BACKGROUND: The objectives of the Dutch National Coordination Center for Travelers' Health Advice (LCR) are to improve the uniformity of travelers' health advice in the Netherlands and to enhance its quality. The LCR offers national guidelines and quality criteria, as well as a telephone consultation service, where health professionals can pose questions regarding travel medicine. Since 2005, a register for qualified travel health professionals has been in place. We studied the quality and relevance of the telephone consultations, to see whether there was a difference between registered as qualified and nonregistered health professionals. METHODS: Telephone questions regarding pretravel advice were logged in September 2007. The questions were categorized as basic or advanced and compared by the profession of the caller, type of institution, and LCR registration of the responsible physician. RESULTS: In 2007, 85% of travel clinic physicians, 42% of general practitioners, and 31% of travel clinic nurses were registered with the LCR. A total of 146 telephone consultations were included in the analysis. Significantly more callers from travel clinics posed advanced questions than those from general practices [odds ratio (OR) 7.6; 95% confidence interval (CI): 3.6-16.1; p= 0.000]. More callers who were registered asked advanced questions, although this difference was not significant (OR 1.7; 95% CI: 0.9-3.3; p= 0.124). Assistants from general practices asked significantly less advanced questions than physicians or nurses. CONCLUSIONS: Opening a register for travel health professionals has led to a large increase of professionals who follow courses and register as travel health professionals. A positive association was found between the quality of the questions and the registration of the responsible physician. The quality of travel health advice given in general practices needs increased attention. PMID- 19674268 TI - Recognizing and reducing the risks of helminthic eosinophilic meningitis in travelers: differential diagnosis, disease management, prevention, and control. PMID- 19674269 TI - Travel and oral anticoagulation. PMID- 19674270 TI - Dead blood under my skin. AB - The diagnostic attitude of western physicians toward migrants' complaints is often an unstable balance between the obstinate search for exotic tropical diseases and the overappreciation of the cultural dimensions of symptoms. Such attitude may divert attention from organic diseases. The careful assessment of all levels of possible misunderstandings (prelinguistic, linguistic, metalinguistic, cultural, and metacultural) may help the physician to discriminate between illness and disease. The long and difficult itinerary leading to the correct diagnosis of congenital myopathy in a migrant from Senegal is described, together with the barriers encountered by the caring staff. PMID- 19674271 TI - Cluster of chikungunya virus infection in travelers returning from Senegal, 2006. AB - Chikungunya virus (CHIKV) infection has been reported in West Africa since 1966, with the last outbreaks from Senegal in 1996 and 1997. We report a cluster of CHIKV infection among travelers returning from Senegal in 2006. Eight imported cases of dengue-like syndrome with fever, joint pain, and skin manifestations were investigated. PMID- 19674272 TI - Chikungunya in Singapore: imported cases among travelers visiting friends and relatives. AB - Chikungunya infections were detected in Singapore among returning travelers who had visited friends and relatives (VFR) in India and Malaysia. These sporadic imported cases occurred over a year before the 2008 chikungunya outbreaks in Singapore, demonstrating the potential for introducing this emerging viral infection into new areas via VFR travel. PMID- 19674273 TI - Eosinophilic meningitis due to Angiostrongylus cantonensis in Germany. AB - We report a case of eosinophilic meningitis due to Angiostrongylus cantonensis in a patient who returned from Thailand. The presence of a compatible epidemiologic history and eosinophilia in cerebrospinal fluid (CSF) lead to the diagnosis, which was confirmed by detection of specific antibodies. After treatment with albendazole and corticosteroids he recovered completely. PMID- 19674274 TI - Appendectomy to remember. PMID- 19674276 TI - Hepatitis B Immunization for all Travelers. PMID- 19674278 TI - Decreased triadin and increased calstabin2 expression in Great Danes with dilated cardiomyopathy. AB - BACKGROUND: Dilated cardiomyopathy (DCM) is a common cardiac disease of Great Dane dogs, yet very little is known about the underlying molecular abnormalities that contribute to disease. OBJECTIVE: Discover a set of genes that are differentially expressed in Great Dane dogs with DCM as a way to identify candidate genes for further study as well as to better understand the molecular abnormalities that underlie the disease. ANIMALS: Three Great Dane dogs with end stage DCM and 3 large breed control dogs. METHODS: Prospective study. Transcriptional activity of 42,869 canine DNA sequences was determined with a canine-specific oligonucleotide microarray. Genome expression patterns of left ventricular tissue samples from affected Great Dane dogs were evaluated by measuring the relative amount of complementary RNA hybridization to the microarray probes and comparing it with expression from large breed dogs with noncardiac disease. RESULTS: Three hundred and twenty-three transcripts were differentially expressed (> or = 2-fold change). The transcript with the greatest degree of upregulation (+61.3-fold) was calstabin2 (FKBP12.6), whereas the transcript with the greatest degree of downregulation (-9.07-fold) was triadin. Calstabin2 and triadin are both regulatory components of the cardiac ryanodine receptor (RyR2) and are critical to normal intracellular Ca2+ release and excitation-contraction coupling. CONCLUSION AND CLINICAL IMPORTANCE: Great Dane dogs with DCM demonstrate abnormal calstabin2 and triadin expression. These changes likely affect Ca2+ flux within cardiac cells and may contribute to the pathophysiology of disease. Microarray-based analysis identifies calstabin2, triadin, and RyR2 function as targets of future study. PMID- 19674279 TI - Size and shape of right heart chambers in mitral valve regurgitation in small breed dogs. AB - BACKGROUND: The contribution of right heart (RH) chamber enlargement to general heart enlargement seen on thoracic radiographs in mitral regurgitation (MR) is not known. OBJECTIVES: To determine the size and shape of the RH chambers in normal dogs and dogs with varying degrees of MR. ANIMALS: Fifty-four privately owned dogs: 13 normal, 41 with varying degrees of MR including 25 with congestive heart failure (CHF). METHODS: Archived first pass radionuclide angiocardiograms were used to produce static images of the RH and left heart (LH) chambers. Indexes of size and shape of the RH and LH chambers were related to severity of MR determined by heart rate-normalized pulmonary transit time (nPTT), vertebral heart scale (VHS), and clinical status. RH shape was measured by a circularity index of RH short axis/long axis. RESULTS: A 2nd degree polynomial fit best described the ratios; RH/LH dimension to nPTT (R(2)= 0.62) and to VHS (R(2)= 0.43), RH/LH area to nPTT (R(2)= 0.64) and to VHS (R(2)= 0.58), all P < .001. RH circularity was decreased in CHF, P < .001. In CHF, the RH chambers of 16 dogs were both flattened and enlarged, whereas 9 had convex septal borders. CONCLUSIONS: RH chambers are not significantly dilated in dogs with mild to moderate MR without CHF. In CHF, RH chambers enlarge and also may be compressed by the LH chambers. Pulmonary hypertension probably is present in some dogs with CHF. Increased sternal contact is not a useful sign of right-sided heart dilatation in MR. PMID- 19674280 TI - A prospective, randomized, double-blinded, placebo-controlled study of human intravenous immunoglobulin for the acute management of presumptive primary immune mediated thrombocytopenia in dogs. AB - BACKGROUND: Immune-mediated thrombocytopenia (IMT) is a common hematologic disorder in dogs. Human intravenous immunoglobulin (hIVIG) may have a beneficial effect in canine IMT. HYPOTHESIS: A single hIVIG infusion (0.5 g/kg) in dogs with presumed primary IMT (pIMT) is a safe adjunctive emergency treatment to accelerate platelet count recovery and shorten hospitalization time without increasing the cost of patient care. ANIMALS: Eighteen client-owned dogs with a presumptive diagnosis of pIMT. METHODS: Prospective, randomized, double-blinded, placebo-controlled clinical trial. RESULTS: There were no identifiable immediate or delayed adverse reactions associated with hIVIG administration over a 6-month period. The median platelet count recovery time for the hIVIG group was 3.5 days (mean + or - SD: 3.7 + or - 1.3 days; range, 2-7 days) and 7.5 days (mean + or - SD: 7.8 + or - 3.9 days; range, 3-12 days) for the placebo group. The median duration of hospitalization for hIVIG group was 4 days (mean + or - SD: 4.2 + or 0.4 days; range, 2-8 days) and 8 days (mean + or - SD: 8.3 + or - 0.6 days; range, 4-12 days) for the placebo group. There was no significant difference between groups with respect to expense of initial patient care, whereas significant reduction in platelet count recovery time (P= .018) and duration of hospitalization (P= .027) were detected in the hIVIG group. CONCLUSIONS AND CLINICAL IMPORTANCE: Compared with corticosteroids alone, adjunctive emergency therapy of a single hIVIG infusion was safe and associated with a significant reduction in platelet count recovery time and duration of hospitalization without increasing the expense of medical care in a small group of dogs with presumed pIMT. PMID- 19674281 TI - Add-on combination therapy with adefovir dipivoxil induces renal impairment in patients with lamivudine-refractory hepatitis B virus. AB - Combination therapy with adefovir dipivoxil (ADV) and lamivudine (LAM) is recommended for patients infected with LAM-refractory hepatitis B virus (HBV). However, the effects of such therapy on renal function and serum phosphorus levels have not been fully evaluated. Combination therapy with ADV and LAM was given to 37 patients infected with LAM-refractory HBV, including 17 with hepatic cirrhosis. Serum HBV DNA levels decreased to below 2.6 log(10) copies/mL in 23 (62%) of 37 patients at 12 months, 25 (78%) of 32 patients at 24 months, and 16 (84%) of 19 patients at 36 months. Except for one cirrhotic patient, serum alanine aminotransferase levels were below 50 IU/L in all patients during combination therapy. Serum creatinine levels increased in 14 (38%) of 37 patients, and serum phosphate levels decreased to below 2.5 mg/mL in 6 (16%) of 37 patients during combination therapy. Patients who received combination therapy for 36 months or longer had a significantly incidence of elevated serum creatinine levels. Fanconi syndrome occurred in a 57-year-old woman with cirrhosis after ADV was added to LAM. Combination therapy with ADV and LAM can maintain biochemical remission in patients with LAM-refractory HBV. However, the dosing interval of ADV should be adjusted according to renal function and serum phosphate levels in patients receiving long-term treatment. PMID- 19674282 TI - Factors associated with rapid and early virologic response to peginterferon alfa 2a/ribavirin treatment in HCV genotype 1 patients representative of the general chronic hepatitis C population. AB - Rapid virologic response (RVR) and complete early virologic response (cEVR) are associated with sustained virologic response to hepatitis C virus (HCV) therapy. We retrospectively examined baseline and on-treatment factors associated with RVR (HCV RNA undetectable at week 4) and cEVR (HCV RNA undetectable at week 12, regardless of week 4 response). The analysis comprised 1550 HCV genotype-1 patients from five clinical trials, including three enriched with difficult-to treat populations, randomized to peginterferon alfa-2a 180 microg/week plus ribavirin 1000-1200 mg/day. Overall, 15.6% achieved RVR and 54.0% achieved cEVR. Baseline factors predictive of RVR were serum HCV RNA 3 x ULN (OR: 2.01; P < 0.0001), non cirrhotic status (OR: 1.92; P = 0.0087), age 13 mg/kg/day was predictive of RVR (OR: 1.69; P = 0.005) and cEVR (OR: 1.24; P = 0.09), whereas peginterferon alfa-2a dose reduction was not. Greater decreases in haematologic parameters were observed in patients who achieved cEVR compared with patients who did not. In conclusion, several baseline and on-treatment factors were associated with RVR and cEVR to peginterferon alfa-2a plus ribavirin in difficult-to-treat HCV genotype-1 patients, providing important prognostic information on the antiviral response in a patient cohort that is reflective of the general chronic hepatitis C population. PMID- 19674283 TI - Hepatic Toll-Like Receptor 3 expression in chronic hepatitis C genotype 1 correlates with treatment response to peginterferon plus ribavirin. AB - Recent studies have shown that enhanced hepatic expression of several innate immune genes predicts non-response to 48 weeks of peginterferon plus ribavirin in chronic hepatitis C genotype 1. This study aimed to further address how gene expression of TLR3/RIG-I signalling correlates with the outcome of the 72-week extended treatment regimen. Relative hepatic mRNA expression and copy numbers of positive- and negative-strand hepatitis C virus (HCV) RNA were determined by real time PCR in 49 patients. Then, a 48-week peginterferon-alpha2b plus ribavirin treatment was commenced and extended to 72 weeks in cases of HCV RNA clearance after week 12. High rate of sustained virologic response was seen both in patients with early HCV clearance (85% [11/13]) and slow virologic responders (85% [11/13]) (per protocol analysis). The response was associated with low TLR3 expression (median, 0.9; range, 0-4.2 vs median, 1.9; range, 0.4-4.9; P = 0.004) but had no relation to the expression of TRIF (P = 0.315), RIG-I (P = 0.953), IPS 1 (P = 0.425), IRF3 (P = 0.329) and interferon-beta (P = 0.584). ROC curve analysis identified TLR3 expression of <1.5 as the best cut-off for predicting response (positive and negative predictive values, 89% [16/18] and 70% [14/20], respectively). The expression was not affected by HCV replication but was higher in female patients (P = 0.043). Multivariate analysis showed TLR3 to be a single baseline predictor (odds ratio 18.5 [95% CI 3.2-111], P = 0.001). Low hepatic TLR3 expression is a novel predictor of response to peginterferon plus ribavirin in genotype 1 patients. PMID- 19674284 TI - Quality of life considerations for patients with chronic hepatitis C. AB - Chronic infection with the hepatitis C virus (HCV) has a profound effect on health-related quality of life (HRQoL) - with fatigue, depression and neurocognitive deficits among the most common complaints. Neuropsychiatric symptoms have prompted research to determine whether the HCV acts within the central nervous system. Replicating virus has been found in central nervous tissues, and changes in neurotransmitter levels in the frontal white matter of patients with chronic hepatitis C are correlated with impaired attention and concentration. Other symptoms of chronic hepatitis C that decrease HRQoL include associated sexual dysfunction and depression. Treatment of chronic HCV infection may temporarily worsen HRQoL, and common adverse effects of currently available agents include fatigue, muscle aches, depression and cognitive deficits. The relationship between sustained viral response and improvement in HRQoL is nonetheless well accepted. Although treatment-related adverse effects may dissuade people from starting therapy and reduce compliance with associated reductions in sustained viral response, for the majority of patients viral clearance produces improvements in both HRQoL and long-term prognosis. Novel agents, with improved adverse effect profiles, may afford more patients the opportunity to achieve a sustained viral response. PMID- 19674286 TI - Community-based hepatitis B screening programs in the United States in 2008. AB - The Centers for Disease Control and Prevention (CDC) recommends hepatitis B surface antigen (HBsAg) testing to identify hepatitis B virus (HBV) infection for foreign-born persons from areas with HBsAg prevalence of > or = 2%. Currently, most HBsAg screening in the United States is performed by independent community organizations. For these HBsAg screening programs, we collected information about the location, number of people screened, other services beyond screening provided, the population/ethnicity groups targeted for screening, and the prevalence of HBsAg among those screened. We identified programs offering screening by contacting programs known to us, from interviews with identified programs, and from structured Internet searches, and collected information using a simple e-mail survey with follow-up phone calls. We identified 55 possible community HBsAg screening programs, of which we successfully contacted 31 programs. In the past year, contacted programs screened an estimated 21 817 patients with an 8.1% average HBsAg prevalence. The majority of programs screened persons born in Asia and their children, and a small number of programs screened persons from Africa or Eastern Europe; very few programs screened U.S.-born persons at risk of HBV infection due to behavioural factors. We identified few or no programs in the American Southeast, the Midwest, and the Southwest outside of California and the Houston area. The HBsAg screening programs that we contacted were effective in identifying and screening patients at risk of HBV as evidenced by the high prevalence observed among those screened. However, their efforts alone are likely insufficient to meet the need for screening recommended by CDC. PMID- 19674285 TI - Early treatment improves outcomes in acute hepatitis C virus infection: a meta analysis. AB - Acute hepatitis C virus infection is associated with high rates of spontaneous clearance and variable rates of treatment-induced clearance. The benefit of early treatment versus awaiting spontaneous clearance is unknown, as is the optimal timing of treatment.We performed a MEDLINE and EMBASE search for the time period 1950 to October 2008. All English language abstracts using the search terms acute hepatitis C, hepatitis C and acute and hepatitis C and acute disease or acute infection were reviewed. Bibliographies were reviewed.Twenty-two studies including 1075 patients met the inclusion criteria. The sustained virologic response (SVR) rate for treated patients was 78%, significantly higher than 55.1% in untreated patients (OR = 3.08, 95% CI: 1.8-4.8 P value <0.0001). Mean time from diagnosis to spontaneous clearance was 9.7 weeks (SD 6.5). SVR rates varied inversely with time from acute HCV diagnosis. SVR rates for treatment within 12 weeks was 82.5% (95% CI: 75.6-89.3), significantly better than the clearance rates in untreated patients (P < 0.001). Response rates fell to 66.9% for treatment between 12 and 24 weeks, and decreased further to 62.5% for treatment beyond 24 weeks. Rates of viral clearance in treated patients with acute hepatitis C virus infection were significantly higher than that in untreated patients. Treatment rates were highest when treatment was initiated within 12 weeks of diagnosis. Based on these findings, we would advocate a 12 week period of observation for spontaneous clearance before treatment initiation. If no clearance has occurred by 12 weeks, treatment should be initiated. PMID- 19674287 TI - The prevalence and clinical correlates of elevated ALT levels in an urban Chinatown community. AB - Alanine aminotransferase (ALT) levels are a primary consideration in treatment decisions regarding hepatitis B virus (HBV) infection. Levels can be elevated as a result of HBV-related liver injury or to other causes. The aim of this study was to assess the background prevalence and clinical correlates of elevated ALT levels in Chinese immigrants who were HBsAg negative. Subjects consisted of adults seen in an urban Chinatown Internal Medicine practice between 1, 2006 and 12, 2007. Clinical information was collected retrospectively. An elevated ALT level was defined as >19 U/L for women and >30 U/L for men. The primary analysis focused on 2457 persons who were HBsAg negative. Their mean age was 55 +/- 18 years, 64% were female and 36% were obese. All subjects were ethnically Chinese and 96% were born in Asia. Thirty percent had an elevated ALT level. In univariate analysis, factors associated with elevated ALT included older age (P < 0.001), female gender (P = 0.001), obesity (P < 0.001), diabetes (P < 0.001), hypercholesterolemia (P < 0.001) and hypertension (P < 0.001). In multivariate analysis BMI, diabetes, female gender and hypercholesterolemia were significantly associated with elevated ALT level. An additional analysis focused on a group of patients who were HBsAg positive, HBeAg negative and had HBV-DNA levels <1000 copies/mL. Fifty percent had elevated ALT levels. In conclusion, elevated ALT levels are common among Chinese immigrants without HBV (30%) and are associated with features of the metabolic syndrome. Liver biopsy should be performed in selected patients with HBV to distinguish the cause of ALT elevation before initiating antiviral therapy. PMID- 19674289 TI - Isolation of Enterobacter cowanii from Eucalyptus showing symptoms of bacterial blight and dieback in Uruguay. AB - AIMS: This study was performed to identify bacterial strains isolated simultaneously with Pantoea species from Eucalyptus trees showing symptoms of bacterial blight and dieback in Uruguay. METHODS AND RESULTS: Several molecular techniques including 16S rRNA and rpoB gene sequencing and DNA-DNA hybridization were used to characterize the gram-negative, facultatively anaerobic, slime producing bacterial strains isolated along with Pantoea species from Eucalyptus. Hypersensitivity reactions (HR) and pathogenicity tests were performed on tobacco and Eucalyptus seedlings, respectively. The isolates clustered closely with the type strain of Enterobacter cowanii in both phylogenetic trees constructed. The DNA-DNA similarity between the isolates and the type strain of Ent. cowanii ranged from 88% to 92%. A positive HR was observed on the tobacco seedlings, but no disease symptoms were visible on the inoculated Eucalyptus seedlings. CONCLUSIONS: Enterobacter cowanii was isolated from trees with symptoms of bacterial blight although strains of this bacterial species do not appear to be the causal agent of the disease. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides the first report of Ent. cowanii isolated from Eucalyptus. Its presence in Eucalyptus tissue suggests that it is an endophyte in trees showing symptoms of blight. PMID- 19674288 TI - Genetic variation in CLDN1 and susceptibility to hepatitis C virus infection. AB - Claudin-1 is a recently discovered co-receptor for hepatitis C virus (HCV) that is required for late-stage binding of the virus. Because variants in the gene that encodes claudin-1 (CLDN1) could play a role in HCV infection, we conducted a 'whole gene association study' among injection drug users (IDUs) to examine whether CLDN1 genetic variants were associated with the risk of HCV infection or with viral clearance. In a cross sectional study, we examined genotype results for 50 single nucleotide polymorphisms (SNPs) across the CLDN1 gene region, comparing genotypes among participants with chronic HCV (n = 658) to those in IDUs who had cleared HCV (n = 199) or remained HCV-uninfected (n = 68). Analyses were controlled for racial ancestry (African-American or European-American) by stratification and logistic regression modeling. We found that participants who remained uninfected more often carried CLDN1 promoter region SNPs -15312C [odds ratio (OR), 1.72; 95% confidence interval (CI) 1.00-2.94; P = 0.048], -7153A (OR, 2.13; 95% CI, 1.25-3.62; P = 0.006) and -5414C (OR, 1.78; 95% CI, 1.06-3.00; P = 0.03). HCV-uninfected participants less often carried CLDN1 IVS1-2983C (OR, 0.55; 95% CI, 0.31-0.97; P = 0.04), which lies in intron 1. CLDN1 -15312C, -7153A and 5414C formed a haplotype in both the African-American and European-American participants and a haplotype analysis supported the association of CLDN1 -7153A in the HCV-uninfected participants. The analyses of HCV clearance revealed no associations with any SNP. These results indicate that genetic variants in regulatory regions of CLDN1 may alter susceptibility to HCV infection. PMID- 19674290 TI - Genetic diversity of gamma-hexachlorocyclohexane-degrading sphingomonads isolated from a single experimental field. AB - AIMS: To isolate gamma-hexachlorocyclohexane (gamma-HCH)-degrading bacteria from a single field and to examine their genetic diversity. METHODS AND RESULTS: Gamma HCH-degrading bacteria were screened from a long-term experimental field in which gamma-HCH has been continuously applied to, and a gamma-HCH-degrading sphingomonad strain SS86 was isolated from in 1986. As the result, five strains of sphingomonads were newly isolated. The sequences of several housekeeping genes separated the six strains, including SS86, into two genotypes. Among the genes involved in gamma-HCH degradation, the sequences of linC, linD and linE were identical among all six strains, that of linA was identical among five strains, and that of linB was diverse. CONCLUSIONS: We calculated that the gamma-HCH degrading populations of the two genotypes arose independently. Not just one but diverse sphingomonads that degrade a particular xenobiotic compound possibly tend to arise and/or accumulate in fields, where that compound has been applied. SIGNIFICANCE AND IMPACT OF THE STUDY: This study indicates the potential usefulness of a long-term continuous application of xenobiotic compounds to an experimental field in that it would potentially generate diverse micro-organisms able to degrade the compounds. PMID- 19674292 TI - Studies on the production of actinomycin-D by Streptomyces griseoruber--a novel source. AB - AIMS: To isolate and characterize bioactive metabolites produced by a micro organism isolated from a soil sample associated with the roots of a medicinal plant, Azadirachta indica. METHODS AND RESULTS: Morphological, cultural, physiological and 16S rRNA homology studies revealed that the organism showed 99% similarity with Streptomyces griseoruber NBRC 12873. One bioactive metabolite (Py2) isolated from the fermented broth was characterized as actinomycin-D (act D). It showed high activity against various gram-positive and gram-negative bacterial cultures, Mycobacterium tuberculosis H37Rv and human neoplastic cells in vitro using standard protocols. CONCLUSIONS: The isolated strain S. griseoruber produced act-D predominantly (210 mg l(-1), c. 88% of the crude) under nonoptimized growth conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: Streptomyces griseoruber may be exploited as a potential source for the commercial production of act-D, as this strain is not reported to produce act-D. Further investigations on the strain for commercial application will be of immense pharmaceutical importance. PMID- 19674293 TI - Genotyping of Edwardsiella ictaluri isolates in Japan using amplified-fragment length polymorphism analysis. AB - AIM: The major objective of the present study was to clarify genetic relationship of isolates of Edwardsiella ictaluri in Japan, which was first found from ayu Plecoglossus altivelis in Japanese rivers in 2007. METHODS AND RESULTS: Ten isolates of Edw. ictaluri in 2007-2008 from ayu and the 1 isolate from bagrid catfish Pelteobagrus nudiceps in Japan were subjected to amplified-fragment length polymorphism (AFLP) analysis. The strains isolated from catfish in United States (ATCC strains) or Indonesia were used as reference strains. The AFLP profiles were all the same among the isolates from Japan, while the polymorphic DNA bands were observed among the strains from United States or Indonesia. The isolates from Japan and Indonesia constituted a genogroup different from the ATCC strains on a dendrogram constructed from the AFLP profiles. CONCLUSION: No DNA polymorphisms were found among Japanese Edw. ictaluri isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: A single clonality of the Edw. ictaluri isolates in Japan suggests the single source of the organism, and the infection in ayu is in the early stage of epidemics. PMID- 19674291 TI - PCR-based assay for the detection of Xanthomonas euvesicatoria causing pepper and tomato bacterial spot. AB - AIMS: To develop a PCR-based assay for Xanthomonas euvesicatoria detection in culture and in planta. METHODS AND RESULTS: A fragment of 1600 bp specific for X. euvesicatoria was found by repetitive extragenic palindromic sequence-PCR. Among the primers designed on the basis of the partially sequenced fragment, the primers Xeu2.4 and Xeu2.5 direct amplification of the expected product (208 bp) for all the X. euvesicatoria strains and not for other related and unrelated phytopathogenic bacteria or saprophytic bacteria isolated from pepper and tomato phyllosphere. The assay permits the detection of X. euvesicatoria in pure culture, with a limit of detection of two bacterial cells and 1 pg of DNA per PCR, and in extracts obtained from asymptomatic inoculated tomato and pepper plants. CONCLUSIONS: Primers Xeu2.4 and Xeu2.5 provide a specific, sensitive and rapid assay for the detection of X. euvesicatoria in culture and in pepper and tomato plants. SIGNIFICANCE AND IMPACT OF THE STUDY: Because X. euvesicatoria is a quarantine organism in the European Union, and it is subjected to stringent international phytosanitary measures, this highly sensitivity PCR-based assay is suitable for its detection in pepper and tomato plant materials to avoid the introduction and spread of the bacterium. PMID- 19674294 TI - Control of microbial contamination of Franz diffusion cell receptor phase in the development of transcutaneous breast cancer therapeutics. AB - AIMS: To determine the micro-organism contamination of excised porcine (pig) ear, and evaluate the use of Cyclopore track-etched membranes (CTEM) for preventing ingress into Franz-type diffusion cells. METHODS: Swabs were taken from four locations and used to inoculate Tryptone Soya Agar (TSA) and Sabouraud Dextrose Agar (SDA) plates. Diffusion cells were assembled to include porcine skin with and without CTEM, and the receptor phase sampled periodically and spread onto plates. RESULTS: Five distinct colony types were isolated after incubation of all swabs on TSA plates at 37 degrees C; on SDA plates, one fungal colony was found at 30 degrees C and one at 37 degrees C. The SDA agar plate incubated at 30 degrees C resulted in the growth of a large diffused white fungal colony. No regional differences were observed. Without the CTEM, the receptor phase became contaminated within 6 h. With the CTEM present, microbial ingress was substantially retarded with visible presumptive fungal growth occurring at 24 h and detectable contamination on both microbiological media at 48 h. CONCLUSIONS: As expected, the native porcine ears were considerably contaminated. The ingress of contamination into the diffusion cell receptor phases can be largely, but not entirely, eliminated using CTEM. The addition of antimicrobial agents was necessary to eliminate micro-organisms that were observed at later time points. SIGNIFICANCE AND IMPACT OF THE STUDY: This article, while highlighting the presence of a high number of micro-organisms on native porcine skin, presents a practical means to reduce the risk of microbial contamination in transdermal/transcutaneous permeation studies, particularly in the study of cell cultures grown within Franz diffusion cell receptor compartments. PMID- 19674295 TI - Induction of purple sulfur bacterial growth in dairy wastewater lagoons by circulation. AB - AIMS: To determine whether circulation of dairy wastewater induces the growth of phototrophic purple sulfur bacteria (PSB). METHODS AND RESULTS: Two dairy wastewater lagoons that were similar in size, geographic location, number and type of cattle loading the lagoons were chosen. The only obvious visual difference between them was that one was stagnant and the water was brown in colour (Farm 1), and the other was circulated and the water was red in colour because of the presence of PSB that contained carotenoid pigments (Farm 2). Both wastewaters were sampled monthly for 3 months and assayed for PSB and extractable carotenoid pigments (ECP). After this point, circulators were placed in the wastewater lagoon on Farm 1, and samples were taken monthly for 9 months and assayed for PSB and ECP. Before the installation of circulators, no PSB-like 16S rRNA sequences or ECP were observed in the wastewater from Farm 1; however, both were observed in the wastewater from Farm 2. After the installation of circulators, statistically greater levels of PSB and extractable carotenoid pigments were observed in the wastewater from Farm 1. CONCLUSIONS: Circulation enhances the growth of PSB in dairy wastewater. SIGNIFICANCE AND IMPACT OF THIS STUDY: Because PSB utilize H(2)S and volatile organic acids (VOA) as an electron source for photosynthesis, and VOA and alcohols as a carbon source for growth, the increase in these bacteria should reduce H(2)S, volatile organic compounds and alcohol emissions from the lagoons, enhancing the air quality in dairy farming areas. PMID- 19674297 TI - Continuity in a longitudinal out-patient attachment for Year 3 medical students. AB - CONTEXT: Concerns about the quality of medical student learning experiences during traditional clerkships have prompted calls to restructure clinical education around continuity. Many US medical schools have added longitudinal out patient attachments to enhance student continuity with patients and supervising doctors. However, continuity with patients can be difficult to achieve and little is known about the independent effect of continuity with a supervising doctor and setting. This study describes students' perceptions of the types of continuity experienced in longitudinal attachments and the learning associated with continuity. METHODS: This is a qualitative study using a grounded theory approach. Interviews were conducted with 12 Year 3 medical students about their continuity experiences with patients, supervisors and settings during their attachment. The resulting data were subjected to thematic analysis. RESULTS: Continuity with supervising doctors provided students with career mentorship and personal support. Student autonomy varied and was most dependent on the supervisor and setting. Students with patient continuity were more likely to report learning about chronic illness and communication skills. Students described the longitudinal attachment as helping them to develop their clinical skills and gain self-confidence within their role as future doctors, and as influencing their career choice. CONCLUSIONS: There is much variation in student experiences of patient continuity during a longitudinal attachment. Continuity with patients, supervisors and settings affects student learning in different ways. Additional dimensions of the experience, such as the nature of the patient doctor relationship, the pace of work and the patient population, impact learning outcomes and should be considered when continuity experiences are being designed. PMID- 19674296 TI - Cultivation of low-temperature (15 degrees C), anaerobic, wastewater treatment granules. AB - AIMS: Anaerobic sludge granules underpin high-rate waste-to-energy bioreactors. Granulation is a microbiological phenomenon involving the self-immobilization of several trophic groups. Low-temperature anaerobic digestion of wastes is of intense interest because of the economic advantages of unheated bioenergy production technologies. However, low-temperature granulation of anaerobic sludge has not yet been demonstrated. The aims of this study were to (i) investigate the feasibility of anaerobic sludge granulation in cold (15 degrees C) bioreactors and (ii) observe the development of methanogenic activity and microbial community structure in developing cold granules. METHODS AND RESULTS: One mesophilic (R1; 37 degrees C) and two low-temperature (R2 and R3, 15 degrees C) laboratory-scale, expanded granular sludge bed bioreactors were seeded with crushed (diameter <0.4 mm) granules and were fed a glucose-based wastewater for 194 days. Bioreactor performance was assessed by chemical oxygen demand removal, biogas production, granule growth and temporal methanogenic activity. Granulation was observed in R2 and R3 (up to 33% of the sludge). Elevated hydrogenotrophic methanogenesis was observed in psychrophilically cultivated biomass, but acetoclastic methanogenic activity was also retained. Denaturing gradient gel electrophoresis of archaeal 16S rRNA gene fragments indicated that a distinct community was associated with developing and mature granules in the low-temperature (LT) bioreactors. CONCLUSIONS: Granulation was observed at 15 degrees C in anaerobic bioreactors and was associated with H(2)/CO(2)-mediated methanogenesis and distinct community structure development. SIGNIFICANCE AND IMPACT OF THE STUDY: Granulation underpins high-rate anaerobic waste treatment bioreactors. Most LT bioreactor trials have employed mesophilic seed sludge, and granulation <20 degrees C was not previously documented. PMID- 19674298 TI - Sociological interpretations of professionalism. AB - CONTEXT: Professionalism is a hot topic in medical education, yet there is debate about what professionalism actually is. The reason is that medical educators primarily frame professionalism as a list of characteristics or behaviours. However, many sociologists of the professions favour more explanatory theories that incorporate political, economic and social dimensions into understanding of the nature and function of professionalism. OBJECTIVES: This paper reviews a range of approaches used in the sociology of the professions to support the argument that medical education needs to reframe its priorities for research into, and the development of, professionalism in medical education. METHODS: The literature on the sociology of the professions was reviewed and summarised in relation to medical education. CONCLUSIONS: A focus on individual characteristics and behaviours alone is insufficient as a basis on which to build further understanding of professionalism and represents a shaky foundation for the development of educational programmes and tools. Contemporary sociological literature on professionalism should have greater prominence in this domain. PMID- 19674299 TI - Historical vicariance and male-mediated gene flow in the toad-headed lizards Phrynocephalus przewalskii. AB - Using mitochondrial and microsatellite DNA data and a population genetic approach, we tested male-mediated gene flow in the toad-headed lizards Phrynocephalus przewalskii. The mitochondrial DNA (ND2 gene), on the one hand, revealed two major lineages and a strong population genetic structure (F(ST) = 0.692; F(ST)' = 0.995). The pairwise differences between the two lineages ranged from 2.1% to 6.4% and the geographical division of the two lineages coincided with a mountain chain consisting of the Helan and Yin Mountains, suggesting a historical vicariant pattern. On the other hand, the nuclear microsatellite DNA revealed a significant but small population genetic structure (F(ST) = 0.017; F(ST)' = 0.372). The pairwise F(ST) among the nine populations examined with seven microsatellite DNA loci ranged from 0.0062 to 0.0266; the assignment test failed to detect any naturally occurring population clusters. Furthermore, the populations demonstrated a weak isolation by distance and a northeast to southwest clinal variation, rather than a vicariant pattern. A historical vicariant event followed by male-mediated gene flow appears to be the best explanation for the data. Approximately 2-5 Ma, climatic change may have created an uninhabitable zone along the Helan-Yin mountain chain and initiated the divergence between the two mitochondrial lineages. With further climatic changes, males were able to disperse across the mountain chain, causing sufficient gene flow that eventually erased the vicariant pattern and drastically reduced the population genetic structure, while females remained philopatric and maintained the mitochondrial DNA (mtDNA) divergence. Although polygyny mating system and female philopatry may partially contribute to the reduced movement of females, other hypotheses, such as female intrasexual aggression, should also be explored. PMID- 19674300 TI - Strong founder effects and low genetic diversity in introduced populations of Coqui frogs. AB - The success of non-native species may depend on the genetic resources maintained through the invasion process. The Coqui (Eleutherodactylus coqui), a frog endemic to Puerto Rico, was introduced to Hawaii in the late 1980s via the horticulture trade, and has become an aggressive invader. To explore whether genetic diversity and population structure changed with the introduction, we assessed individuals from 15 populations across the Hawaiian Islands and 13 populations across Puerto Rico using six to nine polymorphic microsatellite loci and five dorsolateral colour patterns. Allelic richness (R(T)) and gene diversity were significantly higher in Puerto Rico than in Hawaii populations. Hawaii also had fewer colour patterns (two versus three to five per population) than Puerto Rico. We found no isolation by distance in the introduced range, even though it exists in the native range. Results suggest extensive mixing among frog populations across Hawaii, and that their spread has been facilitated by humans. Like previous research, our results suggest that Hawaiian Coquis were founded by individuals from sites around San Juan, but unlike previous research the colour pattern and molecular genetic data (nuclear and mtDNA) support two separate introductions, one on the island of Hawaii and one on Maui. Coquis are successful invaders in Hawaii despite the loss of genetic variation. Future introductions may increase genetic variation and potentially its range. PMID- 19674301 TI - How sympatric is speciation in the Howea palms of Lord Howe Island? AB - The two species of the palm genus Howea (Arecaceae) from Lord Howe Island, a minute volcanic island in the Tasman Sea, are now regarded as one of the most compelling examples of sympatric speciation, although this view is still disputed by some authors. Population genetic and ecological data are necessary to provide a more coherent and comprehensive understanding of this emerging model system. Here, we analyse data on abundance, juvenile recruitment, pollination mode and genetic variation and structure in both species. We find that Howea forsteriana is less abundant than Howea belmoreana. The genetic data based on amplified fragment length polymorphisms markers indicate similar levels of variation in the two species, despite the estimated census population size of H. belmoreana being three times larger than that of H. forsteriana. Genetic structure within species is low although some weak isolation by distance is detectable. Gene flow between species appears to be extremely limited and restricted to early-generation hybrids - only three admixed individuals, classified as F2s or first generation backcrosses to a parental species, were found among sampled palms. We conclude that speciation in Howea was indeed sympatric, although under certain strict definitions it may be called parapatric. PMID- 19674302 TI - Disentangling the effects of historic vs. contemporary landscape structure on population genetic divergence. AB - Increasing habitat fragmentation poses an immediate threat to population viability, as gene flow patterns are changed in these altered landscapes. Patterns of genetic divergence can potentially reveal the impact of these shifts in landscape connectivity. However, divergence patterns not only carry the signature of altered contemporary landscapes, but also historical ones. When considered separately, both recent and historical landscape structure appear to significantly affect connectivity among 51 wood frog (Rana sylvatica) populations. However, by controlling for correlations among landscape structure from multiple time periods, we show that patterns of genetic divergence reflect recent landscape structure as opposed to landscape structure prior to European settlement of the region (before 1850s). At the same time, within-population genetic diversities remain high and a genetic signature of population bottlenecks is lacking. Together, these results suggest that metapopulation processes - not drift-induced divergence associated with strong demographic bottlenecks following habitat loss - underlie the strikingly rapid consequences of temporally shifting landscape structure on these amphibians. We discuss the implications of these results in the context of understanding the role of population demography in the adaptive variation observed in wood frog populations. PMID- 19674303 TI - Genetic structure and evolution of Alpine polyploid complexes: Ranunculus kuepferi (Ranunculaceae) as a case study. AB - The alpine white-flowered buttercup, Ranunculus kuepferi Greuter & Burdet, is a polyploid complex with diploids endemic to the southwestern Alps and polyploids - which have been previously described as apomictic - widespread throughout European mountains. Due to the polymorphic status of both its ploidy level and its reproductive mode, R. kuepferi represents a key species for understanding the evolution of polyploid lineages in alpine habitats. To disentangle the phylogeography of this polyploid taxon, we used cpDNA sequences and AFLP (amplified fragment length polymorphism) markers in 33 populations of R. kuepferi representative of its ploidy level and distribution area. Polyploid individuals were shown to be the result of at least two polyploidization events that may have taken place in the southwestern Alps. From this region, one single main migration of tetraploids colonized the entire Alpine range, the Apennines and Corsica. Genetic recombination among tetraploids was also observed, revealing the facultative nature of the apomictic reproductive mode in R. kuepferi polyploids. Our study shows the contrasting role played by diploid lineages mostly restricted to persistent refugia and by tetraploids, whose dispersal abilities have permitted their range extension all over the previously glaciated Alpine area and throughout neighbouring mountain massifs. PMID- 19674304 TI - Phylogeography of Southern Water Vole (Arvicola sapidus): evidence for refugia within the Iberian glacial refugium? AB - The role of Southern European peninsulas as glacial refugia for temperate species has been widely established, but phylogeographic patterns within refugia are being only recently addressed. Here we describe the phylogeographic patterns for Southern water vole (Arvicola sapidus) in its whole distribution across Iberia and France. Control region and cytochrome b sequences were obtained for 228 samples from 130 localities across Iberia and France. Eighty-five haplotypes were found in total yielding a high overall mitochondrial diversity (pi = 0.027; H = 0.974). Phylogeographic structure was relatively shallow (3.1% average intraspecific divergence) with few supported clades and 95% and 90% maximum parsimony unconnected networks, but significant, as reflected in increased pairwise nucleotide divergences with distance (r = 0.197, P = 0.03) and significant autocorrelation up to approximately 500 km. Spatial analysis of molecular variance analysis detected seven geographical groups explaining 43.73% of the total mitochondrial variation. We detected demographic expansions in three of these groups. A recent colonization of France from Iberia was suggested and estimated around 62 000 years bp by an isolation-with-migration model. Our results suggest the contribution of episodes of isolation in glacial subrefugia in Iberia, but seem to exclude a long-term isolation over successive glacial cycles. Phylogeographic divergence was probably tempered by relatively large population sizes and rapid and extensive mixing among subrefugia during interglacials, that might have eroded the phylogeographic structure accumulated at glacial peaks. Phenotypic differences in A. sapidus do not delineate historically isolated intraspecific divisions and do not warrant subspecific delimitations. Our results do support the existence of subrefugia within Iberia and their role in promoting intraspecific divergences. PMID- 19674305 TI - Allopatric origin of cryptic butterfly species that were discovered feeding on distinct host plants in sympatry. AB - Surveys of tropical insects are increasingly uncovering cryptic species - morphologically similar yet reproductively isolated taxa once thought to comprise a single interbreeding entity. The vast majority of such species are described from a single location. This leaves us with little information on geographic range and intraspecific variation and limits our ability to infer the forces responsible for generating such diversity. For example, in herbivorous and parasitic insects, multiple specialists are often discovered within what were thought to be single more generalized species. Host shifts are likely to have contributed to speciation in these cases. But when and where did those shifts occur, and were they facilitated by geographic isolation? We attempted to answer these questions for two cryptic species within the butterfly Cymothoe egesta that were recently discovered on different host plants in central Cameroon. We first used mtDNA markers to separate individuals collected on the two hosts within Cameroon and then extended our analysis to incorporate individuals collected across the entire pan-Afrotropical range of the original taxon. To our surprise, we found that the species are almost entirely allopatric, dividing the original range and overlapping only in the narrow zone of West-Central Africa where they were first discovered in sympatry. This finding, combined with analyses of genetic variation within each butterfly species, strongly suggests that speciation occurred in allopatry, probably during the Pleistocene. We discuss the implications of our results for understanding speciation among other cryptic species recently discovered in the tropics and argue that more work is needed on geographic patterns and host usage in such taxa. PMID- 19674306 TI - Extreme population genetic differentiation and secondary contact in the freshwater copepod Acanthodiaptomus pacificus in the Japanese Archipelago. AB - We investigated the sequence variation in the mitochondrial cytochrome c oxidase subunit 1 (mtCOI) gene and the nuclear ribosomal internal transcribed spacers (ncITS) of the calanoid copepod Acanthodiaptomus pacificus in Japan. A. pacificus individuals were divided into three divergent mtCOI lineages (mt-A, -B and -C). mt-A was distributed in the northernmost part of Japan, from Hokkaido to the northern part of Honshu Island, whereas mt-C was the southernmost lineage, distributed from central Honshu to Shikoku and Kyushu Islands. mt-B was distributed between these former two lineages, resulting in parapatry with mt-C and mt-A. In all lineages, 80% of the localities were fixed for a single haplotype, and different localities tended to have different haplotypes. The degree of genetic differentiation among these lineages (15-22%) was at an interspecific level, according to the criteria of the DNA barcode technique. However, the topology of ncITS was not congruent with that of mtCOI, as the reciprocal monophyly was not observed within mt-B and mt-C. Therefore, we merged them into the Southern Lineage and separated it from the Northern Lineage (i.e. mt-A). Evidence of introgression was found within the Southern Lineage, while gene flow was not observed between the Northern and Southern Lineages, suggesting that A. pacificus is a cryptic species complex. We also argue that genetic differentiations of A. pacificus in Japan may reflect the history of separation, transgression and regression of the landmass during the formation of current Japanese Archipelago. PMID- 19674307 TI - The influence of distinct pollinators on female and male reproductive success in the Rocky Mountain columbine. AB - Although there are many reasons to expect distinct pollinator types to differentially affect a plant's reproductive success, few studies have directly examined this question. Here, we contrast the impact of two kinds of pollinators on reproductive success via male and female functions in the Rocky Mountain columbine, Aquilegia coerulea. We set up pollinator exclusion treatments in each of three patches where Aquilegia plants were visited by either day pollinators (majority bumble bees), by evening pollinators (hawkmoths), or by both (control). Day pollinators collected pollen and groomed, whereas evening pollinators collected nectar but did not groom. Maternal parents, potential fathers and progeny arrays were genotyped at five microsatellite loci. We estimated female outcrossing rate and counted seeds to measure female reproductive success and used paternity analysis to determine male reproductive success. Our results document that bumble bees frequently moved pollen among patches of plants and that, unlike hawkmoths, pollen moved by bumble bees sired more outcrossed seeds when it remained within a patch as opposed to moving between patches. Pollinator type differentially affected the outcrossing rate but not seed set, the number of outcrossed seeds or overall male reproductive success. Multiple visits to a plant and more frequent visits by bumble bees could help to explain the lack of impact of pollinator type on overall reproductive success. The increase in selfing rate with hawkmoths likely resulted from the abundant pollen available in experimental flowers. Our findings highlighted a new type of pollinator interactions that can benefit a plant species. PMID- 19674308 TI - Testing for local adaptation in Avena barbata: a classic example of ecotypic divergence. AB - Forty years ago, Robert Allard and colleagues documented that the slender wild oat, Avena barbata, occurred in California as two multi-locus allozyme genotypes, associated with mesic and xeric habitats. This is arguably the first example of ecotypes identified by molecular techniques. Despite widespread citation, however, the inference of local adaptation of these ecotypes rested primarily on the allozyme pattern. This study tests for local adaptation of these ecotypes using reciprocal transplant and quantitative trait locus (QTL) mapping techniques. Both ecotypes and 188 recombinant inbred lines (RILs) derived from a cross between them were grown in common garden plots established at two sites representative of the environments in which the ecotypes were first described. Across four growing seasons at each site, three observations consistently emerged. First, despite significant genotype by environment interaction, the mesic ecotype consistently showed higher lifetime reproductive success across all years and sites. Second, the RILs showed no evidence of a trade-off in performance across sites or years, and fitness was positively correlated across environments. Third, at QTL affecting lifetime reproductive success, selection favoured the same allele in all environments. None of these observations are consistent with local adaptation but suggest that a single genotype is selectively favoured at both moist and dry sites. I propose an alternative hypothesis that A. barbata may be an example of contemporary evolution--whereby the favoured genotype is spreading and increasing in frequency--rather than local adaptation. PMID- 19674309 TI - Spatio-temporal population genetic structure of the parasitic mite Spinturnix bechsteini is shaped by its own demography and the social system of its bat host. AB - Information about the population genetic structures of parasites is important for an understanding of parasite transmission pathways and ultimately the co evolution with their hosts. If parasites cannot disperse independently of their hosts, a parasite's population structure will depend upon the host's spatial distribution. Geographical barriers affecting host dispersal can therefore lead to structured parasite populations. However, how the host's social system affects the genetic structure of parasite populations is largely unknown. We used mitochondrial DNA (mtDNA) to describe the spatio-temporal population structure of a contact-transmitted parasitic wing mite (Spinturnix bechsteini) and compared it to that of its social host, the Bechstein's bat (Myotis bechsteinii). We observed no genetic differentiation between mites living on different bats within a colony. This suggests that mites can move freely among bats of the same colony. As expected in case of restricted inter-colony dispersal, we observed a strong genetic differentiation of mites among demographically isolated bat colonies. In contrast, we found a strong genetic turnover between years when we investigated the temporal variation of mite haplotypes within colonies. This can be explained with mite dispersal occuring between colonies and bottlenecks of mite populations within colonies. The observed absence of isolation by distance could be the result from genetic drift and/or from mites dispersing even between remote bat colonies, whose members may meet at mating sites in autumn or in hibernacula in winter. Our data show that the population structure of this parasitic wing mite is influenced by its own demography and the peculiar social system of its bat host. PMID- 19674310 TI - Pleistocene refugia and polytopic replacement of diploids by tetraploids in the Patagonian and Subantarctic plant Hypochaeris incana (Asteraceae, Cichorieae). AB - We report the phylogeographic pattern of the Patagonian and Subantarctic plant Hypochaeris incana endemic to southeastern South America. We applied amplified fragment length polymorphism (AFLP) and chloroplast DNA (cpDNA) analysis to 28 and 32 populations, respectively, throughout its distributional range and assessed ploidy levels using flow cytometry. While cpDNA data suggest repeated or simultaneous parallel colonization of Patagonia and Tierra del Fuego by several haplotypes and/or hybridization, AFLPs reveal three clusters corresponding to geographic regions. The central and northern Patagonian clusters (approximately 38-51 degrees S), which are closer to the outgroup, contain mainly tetraploid, isolated and highly differentiated populations with low genetic diversity. To the contrary, the southern Patagonian and Fuegian cluster (approximately 51-55 degrees S) contains mainly diploid populations with high genetic diversity and connected by high levels of gene flow. The data suggest that H. incana originated at the diploid level in central or northern Patagonia, from where it migrated south. All three areas, northern, central and southern, have similar levels of rare and private AFLP bands, suggesting that all three served as refugia for H. incana during glacial times. In southern Patagonia and Tierra del Fuego, the species seems to have expanded its populational system in postglacial times, when the climate became warmer and more humid. In central and northern Patagonia, the populations seem to have become restricted to favourable sites with increasing temperature and decreasing moisture and there was a parallel replacement of diploids by tetraploids in local populations. PMID- 19674311 TI - Fine-scale comparative phylogeography of a sympatric sister species triplet of subterranean diving beetles from a single calcrete aquifer in Western Australia. AB - Calcrete aquifers in the arid Yilgarn region of central Western Australia are a biodiversity hotspot for stygofauna. A distinct pattern of interspecific size class variation among subterranean dytiscid beetle species has been observed in 29 of these aquifers where either two or three small, medium and/or large sympatric species are found that are in some cases sister species. We used a 3.5 km(2) grid of bores to sample dytiscids on a fine-scale and employed a comparative phylogeographical and population genetic approach to investigate the origins of a sympatric sister species triplet of diving beetles from a single aquifer. Mitochondrial DNA sequence data from the Cytochrome oxidase c subunit I gene revealed that all three species have high levels of haplotype diversity with ancient (approximately 1 million years ago) intra-specific coalescence of haplotypes, but low levels of nucleotide diversity. Population analyses provide evidence for multiple expansion events within each species. There was spatial heterogeneity in the distribution of genetic variation and abundance both within and among the three taxa. Population analyses revealed significant fine-scale differentiation with isolation by distance for Paroster macrosturtensis and P. mesosturtensis, but not the smallest species P. microsturtensis. Haplotype network analyses provided limited or no evidence for past population fragmentation within the large and small species, but substantial historical divergence was observed in P. mesosturtensis that was not spatially structured. A patchy population structure with contemporaneous and historical isolation by distance in the three species is likely to have been a significant isolating and diversifying force, preventing us from ruling out a potential role for allopatric divergence during speciation of this beetle sister triplet. PMID- 19674312 TI - New Zealand phylogeography: evolution on a small continent. AB - New Zealand has long been a conundrum to biogeographers, possessing as it does geophysical and biotic features characteristic of both an island and a continent. This schism is reflected in provocative debate among dispersalist, vicariance biogeographic and panbiogeographic schools. A strong history in biogeography has spawned many hypotheses, which have begun to be addressed by a flood of molecular analyses. The time is now ripe to synthesize these findings on a background of geological and ecological knowledge. It has become increasingly apparent that most of the biota of New Zealand has links with other southern lands (particularly Australia) that are much more recent than the breakup of Gondwana. A compilation of molecular phylogenetic analyses of ca 100 plant and animal groups reveals that only 10% of these are even plausibly of archaic origin dating to the vicariant splitting of Zealandia from Gondwana. Effects of lineage extinction and lack of good calibrations in many cases strongly suggest that the actual proportion is even lower, in keeping with extensive Oligocene inundation of Zealandia. A wide compilation of papers covering phylogeographic structuring of terrestrial, freshwater and marine species shows some patterns emerging. These include: east-west splits across the Southern Alps, east-west splits across North Island, north-south splits across South Island, star phylogenies of southern mountain isolates, spread from northern, central and southern areas of high endemism, and recent recolonization (postvolcanic and anthropogenic). Excepting the last of these, most of these patterns seem to date to late Pliocene, coinciding with the rapid uplift of the Southern Alps. The diversity of New Zealand geological processes (sinking, uplift, tilting, sea level change, erosion, volcanism, glaciation) has produced numerous patterns, making generalizations difficult. Many species maintain pre-Pleistocene lineages, with phylogeographic structuring more similar to the Mediterranean region than northern Europe. This structure reflects the fact that glaciation was far from ubiquitous, despite the topography. Intriguingly, then, origins of the flora and fauna are island-like, whereas phylogeographic structure often reflects continental geological processes. PMID- 19674314 TI - Review article: Low-molecular-weight heparin as an alternative anticoagulant to unfractionated heparin for routine outpatient haemodialysis treatments. AB - Unfractionated heparin is currently the most widely used anticoagulant for outpatient haemodialysis. However, unfractionated heparin is a series of molecules, and as such has variable pharmacodynamics. Low-molecular-weight heparins were developed to improve both drug pharmacokinetic and dynamics, so to provide a reliable predictable clinical effect. The low-molecular-weight heparins are potent agents, but have an increased half-life compared with unfractionated heparin, and also require specialist laboratory monitoring. Despite these apparent drawbacks, low-molecular-weight heparins have become the anticoagulants of choice in Western Europe for routine outpatient haemodialysis sessions, due to the reliability of their clinical effect, and ease of administration, coupled with cost reduction. In standard clinical practice laboratory monitoring is not routinely performed, with drug dosing assessed by clinical inspection of the extracorporeal circuit, and the time for fistula needle sites to stop bleeding. PMID- 19674313 TI - The youngest split in sympatric schizothoracine fish (Cyprinidae) is shaped by ecological adaptations in a Tibetan Plateau glacier lake. AB - Although new empirical evidence shows that sympatric speciation has occurred in some species, there are few indisputable model organisms for this process of speciation. The two subspecies (Gymnocypris eckloni eckloni and G. e. scoliostomus) of the schizothoracine Gymnocypris fish species complex from a small glacier lake in the Tibetan Plateau, Lake Sunmcuo, fit several of the key characteristics of the sympatric speciation model. We used combined mitochondrial control region sequences and the cytochrome b gene (1894 bp) to address the phylogenetics and population genetics of 232 specimens of G. e. eckloni and G. e. scoliostomus, as well as all of its closely related sister species. We found that: (i) a total of four old lineages were uncovered in the widespread G. e. eckloni, of which only one was shown to be shared with all G. e. scoliostomus individuals and (ii) the new subspecies (G. e. scoliostomus) evolved in Lake Sunmcuo from the ancestral G. e. eckloni population within approximately 0.057 Ma. These two taxa of the species complex are morphologically distinct, and reproductive isolation is further suggested. Ecological disruptive selection based on morphological traits (e.g. mouth cleft characters) and food utilization may be a mechanism of incipient speciation of two sympatric populations within Lake Sunmcuo. This study provides the first genetic evidence for sympatric speciation in the schizothoracine fish. PMID- 19674315 TI - Review article: Coagulation cascade and therapeutics update: relevance to nephrology. Part 1: Overview of coagulation, thrombophilias and history of anticoagulants. AB - Coagulation involves the regulated sequence of proteolytic activation of a series of zymogens to achieve appropriate and timely haemostasis in an injured vessel, in an environment that overwhelmingly favours an anticoagulant state. In the non pathological state, the inciting event involves exposure of circulating factor VII/VIIa to extravascularly expressed tissue factor, which brings into motion the series of steps which results in amplification of the initial stimulus, culminating in the conversion of fibrinogen to fibrin and clot formation. The precisely synchronized cascade of events is counter-balanced by a system of anticoagulant mechanisms, which serve to ensure that the haemostatic effect is regulated and does not extend inappropriately. Conversely, in pathological states, these events can escape normal control mechanisms, due to either inherited or acquired defects, which lead to thrombosis. Current anticoagulant therapy, although based on medications that have been in existence for upwards of 80 years, is moving towards targeted therapy for specific coagulation factors and events in the coagulation cascade, based on the current knowledge of the main triggers and key events within the series of reactions that culminates in haemostasis. It remains to be seen whether these newer medications will become first-line therapies for thrombosis in the coming decade. This review aims to elucidate the main events within the coagulation cascade as it is currently understood to operate in vivo, with a brief discussion focusing on hypercoagulable states, and also a short review of the history of anticoagulants as they relate to this model. PMID- 19674317 TI - Conventional nutritional counselling maintains nutritional status of patients on continuous ambulatory peritoneal dialysis in spite of systemic inflammation and decrease of residual renal function. AB - AIM: To evaluate the effect of nutritional counselling on nutritional status in peritoneal dialysis patients. METHODS: Twenty-nine peritoneal dialysis patients were randomly selected to receive conventional nutritional counselling during 6 months of follow up. All patients had monthly clinical and biochemical evaluations, and assessments of dialysis adequacy, inflammation and nutritional status at 0, 3 and 6 months. RESULTS: Moderate-severe malnutrition decreased 28% whereas normal nutrition increased 23% at final evaluation (non-significant). Calorie and protein intake remained stable throughout the study (baseline vs final, calorie: 24 +/- 8 vs 23 +/- 5 Kcal/kg; protein: 1.1 +/- 0.5 vs 1.0 +/- 0.3 g/Kg, respectively). On the other hand, triceps (16 +/- 6 vs 18 +/- 8 mm) and subscapular (17 +/- 8 vs 20 +/- 5 mm) skinfold thicknesses, and mid-arm circumference (27 +/- 3 vs 28 +/- 3 mm) significantly increased; mid-arm muscle area displayed a non-significant trend to increase (30 +/- 9 vs 31 +/- 9 cm(2)) whereas serum albumin significantly increased at the end of study (2.67 +/- 0.46 vs 2.94 +/- 0.48 g/dL). At final evaluation, median renal creatinine clearance decreased (6.3 (0.8-15.3) vs 2.0 (0.1-6.3) L/week per 1.73 m(2)) whereas interleukin-6 increased (2.33 (1.9-7.0) vs 4.02 (2.1-8.4) pg/mL). CONCLUSION: Even though conventional nutritional counselling, as an isolated measure, did not significantly improve all nutritional parameters, it prevented a greater deterioration during 6 months. Nutritional counselling maintained the nutritional status in spite of a decrease in residual renal function and higher systemic inflammation. PMID- 19674316 TI - Cross-sectional audit of blood lead levels in regular outpatient haemodialysis patients dialysing in north London. AB - AIM: Blood lead was measured and found to be high in one haemodialysis patient with atypical abdominal pain and peripheral neuropathy. This prompted an audit of blood lead concentrations in prevalent haemodialysis patients under the care of a University Teaching hospital. METHODS: Blood lead was determined in 271 prevalent adult haemodialysis patients regularly dialysing three times a week, in five dialysis centres and/or on the home dialysis programme. All samples were carefully collected into lead-free plastic containers, and measured by graphite furnace atomic absorption spectrometry. RESULTS: 25.5% of haemodialysis patients had abnormal blood lead concentrations (>200 microg/L), compared with 59% with high-normal values of 100-200 microg/L, and only 15.5% with normal values (<100 microg/L). Blood lead increased with haemodialysis vintage (r = 0.38, P < 0.001), the use of a single carbon filter and reverse osmosis water purification device unit (r = 0.29, P < 0.001), but reduced by urine output (r = -0.44, P < 0.001). CONCLUSION: Despite UK government legislation to reduce permissible lead contamination of drinking water, blood lead was increased in 84.5% of prevalent haemodialysis patients. Single reverse osmosis machines and carbon filters may not be as effective at filtering out possible lead contamination compared with large industrial reverse osmosis devices and carbon filters, and thus blood lead testing should be considered for home haemodialysis patients, particularly in those areas where monochloramines are used to sterilize domestic water supplies. PMID- 19674318 TI - Renal alpha-smooth muscle actin: a new prognostic factor for lupus nephritis. AB - AIM: Systemic lupus erythematosus (SLE) is the prototype of autoimmune disease where renal involvement is frequent and always severe. Histological prognostic factors proposed for lupus nephritis (LN) including the World Health Organization and International Society of Nephrology/Renal Pathology Society--Working Group on the Classification classifications, active (AI) and chronicity (CI) indices may not predict response to treatment. The aim of this study was to correlate alpha smooth muscle actin (alpha-SMA) expression, an early marker of glomerular and interstitial response to injury, to AI and CI, renal scarring progression and response to treatment. METHODS: Fifty-seven kidney biopsy specimens obtained from 32 patients suffering from LN were studied. Twenty patients with class IV LN at first biopsy were identified to study renal progression to chronic renal failure until the use of immunosuppressive treatment. RESULTS: Interstitial alpha-SMA (I alpha-SMA) was correlated only with CI (P < 0.001) whereas mesangial alpha-SMA (M alpha-SMA) was correlated with neither LN activity (P = 0.126) nor sclerosis (P = 0.297). Only I-alpha-SMA was correlated with renal failure (P = 0.01). We divided patients with class IV LN into progressors and non-progressors based on the slope of serum creatinine. At first biopsy, M-alpha-SMA and I-alpha-SMA, but not AI and CI, were correlated with renal failure progression (M-alpha-SMA, 9.7b1.1 vs 7.8b1.4, P = 0.004; and I-alpha-SMA, 9.3b1.1 vs 6.5b3.2, P = 0.011). CONCLUSION: The study data highlight that I-alpha-SMA immunostain in class IV LN patients was correlated with chronicity indices. Moreover, M-alpha-SMA and I-alpha-SMA expression in first biopsy predicted renal progression modality. alpha-SMA expression may therefore be a useful marker to predict renal prognosis in LN. PMID- 19674319 TI - Change of glomerular filtration rate in healthy adults with aging. AB - AIM: In order to determine the relationship between glomerular filtration rate (GFR) and age, the associated factors, and the accurate method of GFR in healthy adults, we conducted a cross-sectional study in community-dwelling adults in Beijing. METHODS: Renal function of 201 clinically healthy subjects was determined using technetium-99 m-labelled diethylene triamine pentacetic acid ((99m)Tc-DTPA). Estimated GFR was calculated with the Cockcroft-Gault (CG) equation, abbreviated Modification of Diet in Renal Disease (MDRD) equation, and plasma clearance of creatinine (Ccr). Serum cystatin C, biomarkers of inflammatory and endothelial cells were analyzed as well. Protein intake, carotid artery intima-media thickness and plaque formation were assayed as well. RESULTS: Glomerular filtration rate was negatively associated with age and the correlation coefficient for (99m)Tc-GFR, CG-GFR, MDRD-GFR, Ccr were -0.643, -0.736, -0.55 and -0.619, respectively (P < 0.001), while the correlation coefficient between cystatin C and age was 0.681 (P < 0.001). Estimated GFR were associated with measured GFR, and the correlation coefficient for Ccr, CG-GFR and MDRD-GFR were 0.813, 0.582 and 0.418, respectively (P < 0.001). The area under the receiver operator curve of Ccr was larger, CG was smaller while MDRD was the smallest, and the difference was significant (P < 0.001). So a predicted equation was presented by cystatin C and C-reactive protein for the elderly. CONCLUSION: In the clinically healthy adults, GFR declined with age. MDRD and CG equation are not suitable to estimate GFR in healthy adults. The predicted equation established by cystatin C and C-reactive protein may be more accurate. PMID- 19674320 TI - Predictors of poor outcomes in steroid therapy for immunoglobulin A nephropathy. AB - BACKGROUND: Steroid therapy appears to be beneficial in patients of immunoglobulin A nephropathy (IgAN), as it causes a reduction in the proteinuria and improves the renal survival. METHODS: A retrospective review of the 5 year follow-up data of 60 patients with IgAN who were treated with steroids was conducted. Steroid non-responders were defined as patients in whom the primary end-point of a 30% decrease of the estimated glomerular filtration rate from baseline was reached. The patients were divided into two groups, namely, the steroid responder group (n = 47) and the steroid non-responder group (n = 13), and the clinicopathophysiological characteristics were compared between the two groups. RESULTS: Significant decrease of the proteinuria was observed in the responder group over the 5 year follow-up period, whereas no significant change of the urinary protein excretion was observed in the non-responder group during the same period. In regard to the pathological findings, significantly higher ratios of glomerular obsolescence and glomerular tuft adhesion to the Bowman's capsule, and significantly higher severity of interstitial fibrosis at the time of diagnosis in the non-responder group than in the responder group were found. The rates of glomerular obsolescence and glomerular tuft adhesion to the Bowman's capsule, the severity of interstitial fibrosis, serum albumin and urinary protein excretion were identified as independent risk factors influencing the rate of renal function deterioration. CONCLUSION: To develop effective therapeutic modalities, it is important to have a thorough understanding of the clinicopathophysiological characteristics of IgAN patients showing poor treatment response to steroids (non-responder group in this study). PMID- 19674322 TI - Late acute antibody mediated rejection 16 years post cadaveric renal transplantation precipitated by recurrent varicella infection. PMID- 19674321 TI - C4d deposition in allograft renal biopsies is an independent risk factor for graft failure. AB - AIM: Association between C4d deposition and renal allograft survival is still uncertain. We retrospectively evaluated the clinical outcome of C4d deposition in allograft renal biopsies. METHODS: One hundred and fifty biopsies from 150 patients with a histological diagnosis of acute rejection from December 1997 to March 2007 were included. Paraffin-embedded sections were stained with a polyclonal antibody using an immunoperoxidase technique. Detailed clinical data were obtained by retrospective review. RESULTS: C4d was stained positively in 74 (49.3%) of 150 cases: 47 (61.5%) biopsies showed diffuse C4d deposition and 27 (38.5%) showed focal C4d deposition. During follow up, significantly more C4d positive patients (24/74 patients, 32.4%) lost their grafts, compared with the C4d-negative group (10/76 patients, 13.2%) (P = 0.005). After a Kaplan-Meier analysis, grafts from the C4d-negative group had a markedly higher survival as compared with the C4d-positive group (P = 0.003, log-rank test). Graft survival among C4d-negative, C4d diffuse-positive, and C4d focal-positive groups was significantly different (P = 0.007, log-rank test). The graft survival rate among C4d-negative patients in early (<6 months) and later biopsies (>6 months), and C4d-positive patients in early and in later biopsies was different (P = 0.028, log-rank test). The adjusted risk ratio of graft failure after Cox proportional hazards multivariate analyses for C4d-positive patients was 3.309 (95% confidence interval, 1.413-6.537; P = 0.004). CONCLUSION: Patients with C4d deposition had an inferior graft survival, especially with diffused C4d deposition, and later experienced acute rejection. C4d deposition was an independent risk factor for graft survival. PMID- 19674323 TI - Aggressive control of secondary hyperparathyroidism can result in immediate efficient blood pressure control in dialysis patients. PMID- 19674324 TI - Changes in arbuscular mycorrhizal fungal phenotypes and genotypes in response to plant species identity and phosphorus concentration. AB - * Arbuscular mycorrhizal fungi (AMF) are plant symbionts that improve floristic diversity and ecosystem productivity. Many AMF species are generalists with wide host ranges. Arbuscular mycorrhizal fungi individuals are heterokaryotic, and AMF populations are genetically diverse. Populations of AMF harbor two levels of genetic diversity on which selection can act, namely among individuals and within individuals. Whether environmental factors alter genetic diversity within populations is still unknown. * Here, we measured genetic changes and changes in fitness-related traits of genetically distinct AMF individuals from one field, grown with different concentrations of available phosphate or different host species. * We found significant genotype-by-environment interactions for AMF fitness traits in response to these treatments. Host identity had a strong effect on the fitness of different AMF, unearthing a specificity of response within Glomus intraradices. Arbuscular mycorrhizal fungi individuals grown in novel environments consistently showed a reduced presence of polymorphic genetic markers, providing some evidence for host or phosphate-induced genetic change in AMF. * Given that AMF individuals can form extensive hyphal networks colonizing different hosts simultaneously, contrasting habitats or soil properties may lead to evolution in the population. Local selection may alter the structure of AMF populations and maintain genetic diversity, potentially even within the hyphal network of one fungus. PMID- 19674325 TI - Selection on grain shattering genes and rates of rice domestication. AB - Molecular cloning of major quantitative trait loci (QTLs) responsible for the reduction of rice grain shattering, a hallmark of cereal domestication, provided opportunities for in-depth investigation of domestication processes. Here, we studied nucleotide variation at the shattering loci, sh4 and qSH1, for cultivated rice, Oryza sativa ssp. indica and Oryza sativa ssp. japonica, and the wild progenitors, Oryza nivara andOryza rufipogon. The nonshattering sh4 allele was fixed in all rice cultivars, with levels of sequence polymorphism significantly reduced in both indica and japonica cultivars relative to the wild progenitors. The sh4 phylogeny together with the neutrality tests and coalescent simulations suggested that sh4 had a single origin and was fixed by artificial selection during the domestication of rice. Selection on qSH1 was not detected in indica and remained unclear in japonica. Selection on sh4 could be strong enough to have driven its fixation in a population of cultivated rice within a period of c. 100 yr. The slow fixation of the nonshattering phenotype observed at the archeological sites might be a result of relatively weak selection on mutations other than sh4 in early rice cultivation. The fixation of sh4 could have been achieved later through strong selection for the optimal phenotype. PMID- 19674326 TI - GintPDX1 encodes a protein involved in vitamin B6 biosynthesis that is up regulated by oxidative stress in the arbuscular mycorrhizal fungus Glomus intraradices. AB - Vitamin B6 is an essential metabolite that has recently been implicated in defense against cellular oxidative stress. In fungi, the de novo biosynthetic pathway of vitamin B6 involves two genes, PDX1 and PDX2. Here, we report a component of the PDX1/PDX2 vitamin B6 biosynthetic pathway in an arbuscular mycorrhizal (AM) fungus. Using rapid amplification of cDNA ends, we isolated the full-length cDNA of a PDX-like gene, GintPDX1, from Glomus intraradices. GintPDX1 expression was analysed by real-time reverse transcription-polymerase chain reaction (RT-PCR). GintPDX1 activity and function were investigated by heterologous complementation of the yeast strainDeltasnz1, which is deficient in vitamin B6 biosynthesis. Sequence data revealed that GintPDX1 is highly homologous to other identified PDX1 proteins. GintPDX1 restores prototrophy to the vitamin B6 auxotrophic yeast mutant and reverts its superoxide sensitivity. GintPDX1 is expressed throughout the fungal life cycle, with the highest transcription levels found in the intraradical fungal structures. GintPDX1 expression was induced in response to hydrogen peroxide, paraquat and copper. The results demonstrate that AM fungi possess at least one component of the machinery necessary for vitamin B6 biosynthesis. Transcriptional regulation of GintPDX1 suggests a role for vitamin B6 as an antioxidant and modulator of reactive oxygen species in G. intraradices. PMID- 19674327 TI - Ontogenetic colour changes in an insular tree species: signalling to extinct browsing birds? AB - * Animals often use colours to hide from predators (crypsis) or advertise defences (aposematism), but there is little evidence for colour-based defence in plants. * Here, we test whether ontogenetic changes in leaf colour of lancewood (Pseudopanax crassifolius) may have been part of a defensive strategy against flightless browsing birds called moa, which were once the only large herbivores in New Zealand. We tested this hypothesis by conducting spectrographic measurements on different-sized plants grown in a common garden. We also compared these results with observations on a closely related, derived species that evolved in the absence of moa on the Chatham Islands. * Spectrographic analyses showed that birds would have difficulty distinguishing seedling leaves against a background of leaf litter. Conversely, brightly coloured tissues flanking spines on sapling leaves are highly conspicuous to birds. Once above the reach of the tallest known moa, adults produce leaves that are typical in appearance to adult leaves. The Chatham Island species lacks ontogenetic colour changes entirely. * Overall, the results indicate that P. crassifolius goes through a remarkable series of colour changes during development, from cryptically coloured seedlings to aposematically coloured saplings, which may have formed a defensive strategy to protect against giant browsing birds. PMID- 19674328 TI - Root-derived CO(2) efflux via xylem stream rivals soil CO(2) efflux. AB - Respiration consumes a large portion of annual gross primary productivity in forest ecosystems and is dominated by belowground metabolism. Here, we present evidence of a previously unaccounted for internal CO(2) flux of large magnitude from tree roots through stems. If this pattern is shown to persist over time and in other forests, it suggests that belowground respiration has been grossly underestimated. Using an experimental Populus deltoides plantation as a model system, we tested the hypothesis that a substantial portion of the CO(2) released from belowground autotrophic respiration remains within tree root systems and is transported aboveground through the xylem stream rather than diffusing into the soil atmosphere. On a daily basis, the amount of CO(2) that moved upward from the root system into the stem via the xylem stream (0.26 mol CO(2) m(-2) d(-1)) rivalled that which diffused from the soil surface to the atmosphere (0.27 mol CO(2) m(-2) d(-1)). We estimated that twice the amount of CO(2) derived from belowground autotrophic respiration entered the xylem stream as diffused into the soil environment. Our observations indicate that belowground autotrophic respiration consumes substantially more carbohydrates than previously recognized and challenge the paradigm that all root-respired CO(2) diffuses into the soil atmosphere. PMID- 19674329 TI - Murray's law, the 'Yarrum' optimum, and the hydraulic architecture of compound leaves. AB - There are two optima for maximizing hydraulic conductance per vasculature volume in plants. Murray's law (ML) predicts the optimal conduit taper for a fixed change in conduit number across branch ranks. The opposite, the Yarrum optimum (YO), predicts the optimal change in conduit number for a fixed taper. We derived the solution for YO and then evaluated compliance with both optima within the xylem of compound leaves, where conduits should have a minimal mechanical role. We sampled leaves from temperate ferns, and tropical and temperate angiosperms Leaf vasculature exhibited greater agreement with ML than YO. Of the 14 comparisons in 13 species, 12 conformed to ML. The clear tendency towards ML indicates that taper is optimized for a constrained conduit number. Conduit number may be constrained by leaflet number, safety requirements, and the fact that the number of conduits is established before their diameter during development. Within a leaf, ML compliance requires leaf-specific conductivity to decrease from petiole to petiolule with the decrease in leaf area supplied. A similar scaling applied across species, indicating lower leaf-specific petiole conductivity in smaller leaves. Small leaf size should offset lower conductivity, and petiole conductance (conductivity/length) may be independent of leaf size. PMID- 19674330 TI - Low-decibel ultrasonic acoustic emissions are temperature-induced and probably have no biotic origin. PMID- 19674331 TI - Novel Phr1 mutations and the evolution of phenol reaction variation in US weedy rice (Oryza sativa). AB - *Red rice, a major agricultural weed, is phenotypically diverse and possesses traits that are similar to both wild and cultivated rice. The genetic resources available for rice make it possible to examine the molecular basis and evolution of traits characterizing this weed. Here, we assess the phenol reaction - a classical trait for distinguishing among cultivated rice varieties - in red rice at the phenotypic and molecular levels. *We phenotyped more than 100 US weed samples for the phenol reaction and sequenced the underlying Phr1 locus in a subset of samples. Data were analyzed in combination with previously published Phr1 data for cultivated rice. *Most weed accessions (96.3%) are positive for the phenol reaction, and samples with a negative response carry loss-of-function alleles that are rare or heretofore undocumented. One such allele may have evolved through mutational convergence of a 1-bp frameshift insertion. Haplotype sharing between red rice and US cultivars suggests occasional crop-weed hybridization. *Our discovery of previously undocumented nonfunctional phr1 alleles suggests that there are likely to be other loss-of-function mutations segregating in Oryza sativa around the world. Red rice may provide a useful study system for understanding the adaptive significance of Phr1 variation in agricultural settings. PMID- 19674332 TI - Expression profiling and functional analysis of Populus WRKY23 reveals a regulatory role in defense. AB - WRKY transcription factors are key regulators that activate and fine-tune stress responses, including defense responses against pathogens. We isolated a poplar (Populus tremulaxPopulus alba) cDNA sequence, PtWRKY23, that encodes the ortholog of Arabidopsis WRKY23 and present the functional analysis of WRKY23, with emphasis on its potential role in resistance to rust infection. To investigate the function of PtWRKY23, we examined PtWRKY23 expression after stress treatments by qRT-PCR and generated PtWRKY23-misexpressing plants. Transgenic plants were assessed for resistance to Melampsora rust and were analyzed using the poplar Affymetrix GeneChip and histological techniques to study the consequences of PtWRKY23 misexpression. PtWRKY23 is rapidly induced by Melampsora infection and elicitor treatments and poplars overexpressing and underexpressing PtWRKY23 were both more susceptible to Melampsora infection than wild type. Transcriptome analysis of PtWRKY23 overexpressors revealed a significant overlap with the Melampsora-infection response. Transcriptome analysis also suggests that PtWRKY23 affects redox homeostasis and cell wall-related metabolism, which was confirmed by analyses that showed that PtWRKY23-misexpressing plants have altered peroxidase activity, apparent H(2)O(2) accumulation and lignin deposition. Our results show that PtWRKY23 affects resistance to Melampsora infection and that this may be caused by deregulation of genes that disrupt redox homeostasis and cell wall metabolism. PMID- 19674334 TI - Are species shade and drought tolerance reflected in leaf-level structural and functional differentiation in Northern Hemisphere temperate woody flora? AB - Leaf-level determinants of species environmental stress tolerance are still poorly understood. Here, we explored dependencies of species shade (T(shade)) and drought (T(drought)) tolerance scores on key leaf structural and functional traits in 339 Northern Hemisphere temperate woody species. In general, T(shade) was positively associated with leaf life-span (L(L)), and negatively with leaf dry mass (M(A)), nitrogen content (N(A)), and photosynthetic capacity (A(A)) per area, while opposite relationships were observed with drought tolerance. Different trait combinations responsible for T(shade) and T(drought) were observed among the key plant functional types: deciduous and evergreen broadleaves and evergreen conifers. According to principal component analysis, resource-conserving species with low N content and photosynthetic capacity, and high L(L) and M(A), had higher T(drought), consistent with the general stress tolerance strategy, whereas variation in T(shade) did not concur with the postulated stress tolerance strategy. As drought and shade often interact in natural communities, reverse effects of foliar traits on these key environmental stress tolerances demonstrate that species niche differentiation is inherently constrained in temperate woody species. Different combinations of traits among key plant functional types further explain the contrasting bivariate correlations often observed in studies seeking functional explanation of variation in species environmental tolerances. PMID- 19674333 TI - Hydraulic adjustment of Scots pine across Europe. AB - * The variability of branch-level hydraulic properties was assessed across 12 Scots pine populations covering a wide range of environmental conditions, including some of the southernmost populations of the species. The aims were to relate this variability to differences in climate, and to study the potential tradeoffs between traits. * Traits measured included wood density, radial growth, xylem anatomy, sapwood- and leaf-specific hydraulic conductivity (K(S) and K(L)), vulnerability to embolism, leaf-to-sapwood area ratio (A(L) : A(S)), needle carbon isotope discrimination (Delta13C) and nitrogen content, and specific leaf area. * Between-population variability was high for most of the hydraulic traits studied, but it was directly associated with climate dryness (defined as a combination of atmospheric moisture demand and availability) only for A(L) : A(S), K(L) and Delta13C. Shoot radial growth and A(L) : A(S) declined with stand development, which is consistent with a strategy to avoid exceedingly low water potentials as tree size increases. In addition, we did not find evidence at the intraspecific level of some associations between hydraulic traits that have been commonly reported across species. * The adjustment of Scots pine's hydraulic system to local climatic conditions occurred primarily through modifications of A(L) : A(S) and direct stomatal control, whereas intraspecific variation in vulnerability to embolism and leaf physiology appears to be limited. PMID- 19674335 TI - Computational analysis of flowering in pea (Pisum sativum). AB - During plant development, the transition from a vegetative to reproductive state is a critical event. For decades, pea (Pisum sativum) has been used as a model species to study this transition. These studies have led to a conceptual, qualitative model for the control of flower initiation, referred to as the 'classical' model. This model involves many inputs, namely photoperiod, genetic states and two mobile signals which interact to determine the first node of flowering. Here, we developed a computational model based on the hypotheses of the classical model. Accordingly, we converted qualitative hypotheses into quantitative rules. We found that new hypotheses, in addition to those already described for the classical model, were required that explicitly described the signals. In particular, we hypothesized that the key flowering gene HR interacts with the photoperiod pathway to control flowering. The computational model was tested against a wide range of biological data, including pre-existing and new experimental results presented here, and was found to be accurate. This computational model, together with ongoing experimental advances, will assist future modelling efforts to increase our understanding of flowering in pea. PMID- 19674337 TI - Massively parallel 454 sequencing indicates hyperdiverse fungal communities in temperate Quercus macrocarpa phyllosphere. AB - * This study targeted the fungal communities in the phyllosphere of Quercus macrocarpa and compared the fungal species richness, diversity and community composition among trees located within and outside a small urban center using recently developed 454 sequencing and DNA tagging. * The results indicate that the fungal phyllosphere communities are extremely diverse and strongly dominated by ascomycetes, with Microsphaeropsis [two Operational Taxonomic Units (OTUs); 23.6%], Alternaria (six OTUs; 16.1%), Epicoccum (one OTU; 6.0%) and Erysiphe (two OTUs; 5.9%) as the most abundant genera. * Although the sequencing effort averaged 1000 reads per tree and detected nearly 700 distinct molecular OTUs at 95% internal transcribed spacer 1 similarity, the richness of the hyperdiverse phyllosphere communities could not be reliably estimated as nearly one-half of the molecular OTUs were singletons. * The fungal communities within and outside the urban center differed in richness and diversity, which were lower within the urban development. The two land-use types contained communities that were distinct and more than 10% of the molecular OTUs differed in their frequency. PMID- 19674336 TI - Impact of CCR1 silencing on the assembly of lignified secondary walls in Arabidopsis thaliana. AB - A cinnamoyl-CoA reductase 1 knockout mutant in Arabidopsis thaliana was investigated for the consequences of lignin synthesis perturbation on the assembly of the cell walls. The mutant displayed a dwarf phenotype and a strong collapse of its xylem vessels corresponding to lower lignin content and a loss of lignin units of the noncondensed type. Transmission electron microscopy revealed that the transformation considerably impaired the capacity of interfascicular fibers and vascular bundles to complete the assembly of cellulose microfibrils in the S(2) layer, the S(1) layer remaining unaltered. Such disorder in cellulose was correlated with X-ray diffraction showing altered organization. Semi quantitative immunolabeling of lignins showed that the patterns of distribution were differentially affected in interfascicular fibers and vascular bundles, pointing to the importance of noncondensed lignin structures for the assembly of a coherent secondary wall. The use of laser capture microdissection combined with the microanalysis of lignins and polysaccharides allowed these polymers to be characterized into specific cell types. Wild-type A. thaliana displayed a two fold higher syringyl to guaiacyl ratio in interfascicular fibers compared with vascular bundles, whereas this difference was less marked in the cinnamoyl-CoA reductase 1 knockout mutant. PMID- 19674338 TI - A look inside: localization patterns and functions of intracellular plant aquaporins. AB - Aquaporins form a superfamily of intrinsic channel proteins in the plasma and intracellular membranes of plant cells. While a lot of research effort has substantiated the importance of plasma membrane aquaporins for the regulation of plant water homeostasis, comparably little is known about the function of intracellular aquaporins. Yet, various low-molecular-weight compounds, in addition to water, were recently shown to permeate some of these aquaporins. In this review, we examine the diversity of transport properties and localization patterns of intracellular aquaporins. The discussed profiles include, for example, water and ammonia transport across the tonoplast or CO2 transport through the chloroplast envelope. Furthermore, we try to assess to what extent the diverse aquaporin distribution patterns, in relation to the high degree of compartmentation of plant cells, can be linked to a wide range of cellular functions. PMID- 19674339 TI - An actinoporin plays a key role in water stress in the moss Physcomitrella patens. AB - * Modern land plants arose from a green algae-like ancestor c. 480 million years ago. While several novel morphological features were critical for survival in the aerial environment, physiological innovation undoubtedly played a key role in the colonization of terrestrial habitats. Recently, actinoporin genes, a small group of pore-forming toxins from sea anemones, have been found in the bryophyte and lycophyte lineages of land plants where they are upregulated in water-stressed tissues. * The bryoporin gene in the moss Physcomitrella patens (PpBP) was functionally characterized by RNA blot analyses and overexpression in P. patens. In order to examine functional homology between PpBP and sea anemone actinoporins, the recombinant PpBP was subjected to hemolytic analysis of pig blood cells, which is one of the specific activities of actinoporins. * PpBP was upregulated by various abiotic stresses, in particular most strongly by dehydration stress. Overexpression of the bryoporin gene heightens drought tolerance in P. patens significantly. In addition, PpBP shared the highest structural homology with actinoporins in a three-dimensional structural database and showed hemolytic activity. * These results suggest that this phylogenetic distribution may have resulted from an ancient horizontal gene transfer and actinoporins may have played an important role in early land plants. PMID- 19674340 TI - Electron transfer mediators and other metabolites and cofactors in the treatment of mitochondrial dysfunction. AB - Mitochondrial disorders (MDs) are caused by impairment of the mitochondrial electron transport chain (ETC). The ETC is needed for oxidative phosphorylation, which provides the cell with the most efficient energy outcome in terms of ATP production. One of the pathogenic mechanisms of MDs is the accumulation of reactive oxygen species. Mitochondrial dysfunction and oxidative stress appear to also have a strong impact on the pathogenesis of neurodegenerative diseases and cancer. The treatment of MDs is still inadequate. Therapies that have been attempted include ETC cofactors, other metabolites secondarily decreased in MDs, antioxidants, and agents acting on lactic acidosis. However, the role of these dietary supplements in the treatment of the majority of MDs remains unclear. This article reviews the rationale for their use and their role in clinical practice in the context of MDs and other disorders involving mitochondrial dysfunction. PMID- 19674342 TI - Influence of dietary gangliosides on neonatal brain development. AB - Gangliosides are sialic acid-containing glycosphingolipids. Gangliosides are found in human milk; understanding of the potential role of gangliosides in infant development is emerging, with suggested roles in the brain and gut. Ganglioside accretion in the developing brain is highest in utero and in early neonatal life, during the periods of dendritic branching and new synapse formation. Further, brain contains the highest relative ganglioside content in the body, particularly in neuronal cell membranes concentrated in the area of the synaptic membrane. Gangliosides are known to play a role in neuronal growth, migration and maturation, neuritogenesis, synaptogenesis, and myelination. In addition to their roles in development and structure of the brain, gangliosides also play a functional role in nerve cell communication. It is less well known whether dietary gangliosides can influence the development of cognitive function. This review summarizes current knowledge on the role gangliosides play in brain development. PMID- 19674341 TI - Mendelian randomization in nutritional epidemiology. AB - Nutritional epidemiology aims to identify dietary and lifestyle causes for human diseases. Causality inference in nutritional epidemiology is largely based on evidence from studies of observational design, and may be distorted by unmeasured or residual confounding and reverse causation. Mendelian randomization is a recently developed methodology that combines genetic and classical epidemiological analysis to infer causality for environmental exposures, based on the principle of Mendel's law of independent assortment. Mendelian randomization uses genetic variants as proxies for environmental exposures of interest. Associations derived from Mendelian randomization analysis are less likely to be affected by confounding and reverse causation. During the past 5 years, a body of studies examined the causal effects of diet/lifestyle factors and biomarkers on a variety of diseases. The Mendelian randomization approach also holds considerable promise in the study of intrauterine influences on offspring health outcomes. However, the application of Mendelian randomization in nutritional epidemiology has some limitations. PMID- 19674343 TI - Does tailoring make a difference? A systematic review of the long-term effectiveness of tailored nutrition education for adults. AB - Tailoring individualizes information to the receiver and provides a potential strategy for improving dietary intakes. The present systematic review summarizes evidence for the long-term (> or =6 months) effectiveness of tailored nutrition education for adults and includes priority population groups. Key electronic databases and relevant bibliographies were searched for trials measuring the following outcomes: nutrition-related health behaviors (e.g., dietary intake and food purchases) and anthropometric measures. Data synthesis was comprised of meta analysis (for 15 trials including all population groups) and narrative review (for five trials of priority population groups). Overall, the quality of the studies was moderate to good. Tailored nutrition education was found to be a promising strategy for improving the diets of adults (including those in priority population groups) over the long term. However, future studies should ensure adequate reporting of research design and methods and reduce the chances of false positive findings by using more objective measures of diet, clearly identifying the primary study outcome, and concentrating on outcomes most relevant to nutrition-related disease. PMID- 19674346 TI - Polymorphisms in inflammation genes, tobacco smoke and furred pets and wheeze in children. AB - Persistent wheeze in childhood is associated with airway inflammation. The present study investigated relationships between polymorphisms in inflammatory genes, exposure to tobacco smoke and furred pets and risk of recurrent wheeze in children. Within a birth cohort of 101,042 children we identified 1111 eighteen month old cases with recurrent wheeze and 735 wheeze-free controls among 11942 children recruited in the Copenhagen area. Polymorphisms in IL-4R, IL-8, IL-13, SPINK5, and CD14 were genotyped. Interviews at gestational wks 12 and 30, and at age 6 and 18 months included questions on number of episodes with wheeze (18 months), exposure to tobacco smoke and pet-keeping. Recurrent wheeze was defined as at least four episodes of wheeze before the child was 18 months old. There was a statistically significant association between the IL-13 Arg144Gln polymorphism and risk of recurrent wheeze (p = 0.01). Furthermore, there was a statistically significant interaction between this polymorphism and exposure to tobacco smoke during pregnancy, though this was probably a chance finding. There were no other statistically significant effects of the polymorphisms or interactions with exposure to tobacco smoke in relation to the risk of recurrent wheeze. Polymorphisms in IL-8 affected the association between pet-keeping and risk of wheeze. Polymorphisms in inflammation genes might affect the association between environmental exposures and risk of recurrent wheeze in early childhood. PMID- 19674347 TI - Parasites and allergy. PMID- 19674344 TI - Vitamin D and the occurrence of depression: causal association or circumstantial evidence? AB - While recent laboratory-based studies have substantially advanced our understanding of the action of vitamin D in the brain, much is still unknown concerning how vitamin D relates to mood. The few epidemiological studies of vitamin D and depression have produced inconsistent results and generally have had substantial methodological limitations. Recent findings from a randomized trial suggest that high doses of supplemental vitamin D may improve mild depressive symptoms, but important questions persist concerning how vitamin D may affect monoamine function and hypothalamic-pituitary-adrenal axis response to stress, whether vitamin D supplementation can improve mood in individuals with moderate-to-severe depression, and whether vitamin D sufficiency is protective against incident depression and recurrence. At this time, it is premature to conclude that vitamin D status is related to the occurrence of depression. Additional prospective studies of this relationship are essential. PMID- 19674348 TI - In this issue. Volume 20 Issue 5 (August 2009). PMID- 19674349 TI - Shellfish allergy in children. AB - Food allergies affect approximately 3.5-4.0% of the world's population and can range from a mere inconvenience to a life-threatening condition. Over 90% of food allergies in childhood are caused by eight foods: cow's milk, hen's egg, soy, peanuts, tree nuts, wheat, fish, and shellfish. Shellfish allergy is known to be common and persistent in adults, and is an important cause of food induced anaphylaxis around the world for both children and adults. Most shellfish allergic children have sensitivity to dust mite and cockroach allergens. Diagnostic cut-off levels for skin prick testing in children with shrimp allergy exist but there are no diagnostic serum-specific immunoglobulin E (IgE) values. All patients with symptoms of IgE-mediated reactions to shellfish should receive epinephrine autoinjectors, even if the initial symptoms are mild. In this study, we review three cases of clinical presentations of shellfish allergy in children. PMID- 19674350 TI - Association of passive exposure of pregnant women to environmental tobacco smoke with asthma symptoms in children. AB - Environmental tobacco smoke (ETS) is a significant risk factor for the presence and increased severity of asthma- and allergy-related symptoms in children. Smoking during pregnancy has detrimental effects on asthma-associated outcomes in childhood. Whether passive exposure of pregnant women to ETS may also lead to asthma in their offspring, is not known. The aim of this study was to investigate the association of passive exposure of pregnant women to ETS and asthma- and/or allergy-related symptoms in Preschool children. Cross-sectional data were collected with questionnaires from 2374 Preschool children, recruited from public and private nurseries and day-care centers. Parental smoking was significantly associated with wheezing symptoms in their children. Mother's active smoking during pregnancy significantly increased the risk for occurrence of asthma symptoms and/or medically diagnosed asthma in Preschool children in a dose dependent manner. Passive exposure to ETS, mainly during the third trimester of pregnancy, was significantly associated with asthma- and allergy-related symptoms after adjusting for several confounders in a multivariate analysis (current wheeze: OR = 1.42, 95% CI = 1.06-1.91, pruritic rash ever: OR= 1.45, 95% CI = 1.01-2.08). Passive exposure of pregnant women to ETS during the third trimester is positively associated with asthma- and allergy-related symptoms in their Preschool age children. Public health policies should be oriented not only towards smoking cessation, but also reinforce elimination of ETS exposure of pregnant women. PMID- 19674351 TI - High mother's educational level: an associated factor for wheezing infants? PMID- 19674352 TI - Levels of Th2 chemokine CCL27 in adolescents: no impact of obesity. PMID- 19674353 TI - Early postpartum discharge and breastfeeding: an observational study from France. AB - BACKGROUND: Early postpartum discharge is a recent practice in France, but the influence of a shortened hospital stay on subsequent breastfeeding is unknown. The objective of the present study was to compare the breastfeeding mode after early discharge (ED) and conventional discharge (CD) from a hospital maternity unit. METHODS: An observational study was conducted in a French university hospital among 135 breastfeeding mothers, who delivered between 1 January and 31 July 2006. Forty-five ED mothers were matched with 90 CD mothers on 13 criteria. A structured questionnaire was used to collect data regarding feeding practices at 10 weeks postpartum, the period corresponding to paid maternity leave. RESULTS: Exclusive breast-, mixed, and bottle feedings were reported by, respectively, 35 (77.8%), three (6.7%) and seven (15.5%) ED mothers and 64 (71.1%), 12 (13.3%) and 14 (15.6%) CD mothers (no significant differences). Satisfaction with support for breastfeeding and reasons for switching to mixed or bottle feeding were comparable in the two groups. Multivariate analysis indicated that only the planned duration of breastfeeding and the mother's dissatisfaction with help significantly influenced breastfeeding prevalence. CONCLUSIONS: Early postpartum hospital discharge organized by skilled professionals is compatible with a satisfactory rate of exclusive breastfeeding up to the return to work. Formalized programs of instruction for perinatal professionals would help to reduce early abandonment. PMID- 19674354 TI - Nebulized hypertonic saline/salbutamol solution treatment in hospitalized children with mild to moderate bronchiolitis. AB - BACKGROUND: The objective of this study was to determine the efficacy and safety of nebulized 3% hypertonic saline solution and salbutamol in the treatment of mild to moderate bronchiolitis. METHODS: In a randomized controlled trial, 93 infants with mild to moderate bronchiolitis were divided into two groups. The infants received inhalation of 2.5 mg (0.5 mL) salbutamol dissolved in either 4.0 mL normal (0.9%) saline (control group, n= 43) or 4.0 mL hypertonic (3%) saline (treatment group, n= 50). The therapy was repeated three times daily until discharge. Cough, wheezing, pulmonary physical signs, and the length of hospital stay were recorded. RESULTS: Wheezing remission time was 3.8 + or - 1.1 days in the control group and 2.7 + or - 0.9 days in the treatment group (P < 0.01). Cough remission time was 6.3 + or - 0.9 days in the control group and 5.3 + or - 0.8 days in the treatment group (P < 0.01). The moist crackles disappeared at 5.4 + or - 0.8 days in the treatment group versus 6.2 + or - 0.9 days in the control group (P < 0.01). Furthermore, the average length of hospital stay decreased from 7.4 + or - 1.5 days in the control group to 6.0 + or - 1.2 days in the treatment group (P < 0.01). No obvious adverse effects were observed. CONCLUSIONS: Inhalation of nebulized 3% hypertonic saline solution and salbutamol is a safe and effective therapy for patients with mild to moderate bronchiolitis. PMID- 19674355 TI - The effect of low-carbohydrate diet on left ventricular diastolic function in obese children. AB - BACKGROUND: This study was conducted to evaluate left ventricle (LV) functions using conventional and tissue Doppler imaging in childhood obesity and to identify the effects of diet on LV diastolic functions. METHODS: Conventional and tissue Doppler echocardiographic measurements were compared in 34 obese children and 24 age- and gender-matched lean controls. Fasting plasma glucose, insulin and homeostatic model assessment of insulin resistance levels were also obtained. Thirty-one of the obese children were subjected to a low-carbohydrate diet and their follow-up measurements were obtained after 6 months. RESULTS: Left atrial diameter, LV mass and LV mass index were higher in obese children than in lean controls. Lateral mitral myocardial early diastolic (E(m)) and peak E(m)/myocardial late diastolic (A(m)) were lower, and mitral E/E(m) and lateral mitral myocardial isovolumetric relaxation time were higher in obese subjects than in lean controls. Insulin and homeostatic model assessment of insulin resistance levels were higher in obese patients and decreased significantly after diet. After diet therapy, lateral mitral E(m) and peak E(m)/A(m), were increased, mitral E/E(m) and myocardial isovolumetric relaxation time were decreased. CONCLUSIONS: Obesity predisposes children to increased preload reserve, left ventricular subclinical diastolic dysfunction and deterioration in diastolic filling. Weight reduction with a low-carbohydrate diet seems to be associated with significant improvement in LV diastolic function and a decrease in diastolic filling, as well as causing reversal in insulin resistance seen in obese children. PMID- 19674356 TI - Neutropenia in the acute phase of Kawasaki disease and prevention of coronary artery aneurysm. AB - BACKGROUND: The significance of neutropenia in Kawasaki disease (KD) has not been fully elucidated as yet. METHODS: Subjects were retrospectively sampled from two clinical trials. These patients treated with aspirin alone (ASA) and PolyglobinN Bayel (PolyN) given as i.v. immunoglobulin were categorized as ASA-early (n = 0), ASA-late (n = 8), PolyN-early (n = 18), or PolyN-late (n = 27) based on the therapy administered and the incidence of neutropenia before the 10th day of illness (DI) and after 11 DI. Data regarding the time of onset of neutropenia, and incidence of coronary artery lesion (CAL) formation were obtained. P < 0.05 was considered statistically significant. RESULTS: No patients in the ASA group exhibited neutropenia within 10 DI. The time of onset of neutropenia in the PolyN early group was 8 +/- 1.3 DI. That in the PolyN-late group (19.8 +/- 8 DI) was earlier than in the ASA-late group (26.6 +/- 14 DI; P < 0.025). PolyN-early patients had a lower incidence of CAL formation than ASA-non patients (patients without neutropenia in the ASA group; P = 0.00019) and ASA-late patients (P = 0.04). That in the PolyN-early group tended to be lower than in the PolyN-late group (P < 0.1). CONCLUSION: Early neutropenia indicated that circulating neutrophils within 10 DI may play an indispensable role in the following sequence to CAL formation in KD. PMID- 19674357 TI - Turner syndrome phalangeal screening based on a two-stage linear regression concept. AB - BACKGROUND: Turner syndrome (TS) is a congenital chromosomal abnormality, resulting in short stature, short fourth metacarpal, and retarded skeletal maturation in children. The existing methods of diagnosis, which include carpal angle, metacarpal sign, and body mass index (BMI), cannot accurately diagnose TS. The authors propose a novel procedure for examining the hand skeleton to distinguish between normal individuals and patients with TS. METHODS: This investigation was divided into two parts. In the first part, existing methods (evaluation of the metacarpal sign, measurement of the carpal angle, and determination of BMI) were used. Examination in the second part was based on the two-stage screening method (TSSM). In the first stage in TSSM, the ratio of the lengths of the distal-middle phalanges of the fifth digit was determined in normal subjects with average range of satisfactory body height and TS patients. A suitable cut-off was found on linear regression and used to divide the plot into TS patients and normal subjects. In the second stage, the normal section was transferred to another group based on bone and chronological ages. A greater number of patients were diagnosed with TS using this method. Finally, four cut off parameters were determined on linear regression analysis. Results with optimal sensitivity and specificity were automatically obtained. RESULTS: The combination of TSSM with optimal programming (sensitivity = 0.81 and specificity = 0.91) was satisfactory for diagnosing TS patients. CONCLUSION: TSSM can suitably evaluate growth of the hand skeleton to distinguish between normal individuals and patients with TS. PMID- 19674358 TI - Diagnostic spectrum of congenital hypothyroidism in Turkish children. AB - BACKGROUND: Congenital hypothyroidism (CH) is classified as transient or permanent. Transient CH can be treated with short-term thyroxine replacement therapy or followed up without therapy, whereas lifelong thyroxine replacement is needed in permanent CH. Determination of the underlying etiology is essential for determination of follow-up strategy. The purpose of the present study was therefore to assess the role of color Doppler ultrasonography (CDU) in etiologic diagnosis of CH together with radionuclide method and grayscale ultrasonography (GSU). METHODS: A total of 182 patients (83 female, 99 male) were evaluated. To determine etiologic diagnosis, the patients underwent a free T4 (fT4), thyroid stimulating hormone (TSH) and urinary iodine level measurement, and thyroid scintigraphy, perchlorate discharge test, CDU, and GSU. RESULTS: Fifty-four patients had transient and 97 had permanent CH. Isolated hyperthyrotropinemia was diagnosed in 31 patients. Transient CH was due to iodine deficiency in 22 and excess iodine in 13 patients. In 97 patients with permanent CH, ectopia was present in 32 patients and agenesis of the thyroid gland was found in 22 patients, while 43 were diagnosed with dyshormonogenesis. In the ectopia group, GSU failed to detect ectopic tissues in all cases, whereas CDU was successful in determining the presence of ectopic thyroid in 20 cases. The sensitivity of CDU was 80% in determining ectopic tissue. CONCLUSION: To determine etiologic diagnosis, radionuclide methods and sonographic modalities should be assessed together. The gold standard in the diagnosis of ectopic thyroid tissue is thyroid scintigraphy. CDU may be a major supportive diagnostic tool in the evaluation of ectopic thyroid gland. PMID- 19674359 TI - How frequent is adenoid obstruction? Impact on the diagnostic approach. AB - BACKGROUND: The rate of adenoidectomy has increased over the past years. The initial assessment methods are sometimes overused. The aims of the present study were to evaluate the use of these methods, estimate the incidence of obstructive adenoid and refine the approach to this problem. METHODS: This is a prospective observational study. All children referred for chronic nasal obstruction had anterior rhinoscopy and a routine physical examination. The palatal airway was assessed on lateral nasopharyngeal roentgenograms when these were requested. The degree of obstruction was measured on endoscopy when performed. The efficacy of each method was evaluated, as well as its necessity. The incidence of obstructive adenoid was calculated. RESULTS: A total of 213 patients aged 6 months-13 years (mean 5.1 years, median 4 years) were enrolled. One hundred patients had radiological evaluation while 65 had endoscopy during their initial assessment. Endoscopy was the most efficacious in reaching a proper diagnosis (100%) followed by clinical assessment (84.2%), and radiology (75%). The necessity of endoscopy, however, was 63.1% and that of radiology 63% within their respective groups. The incidence of obstructive adenoid was 57.7% (1.6% choanal). Clinical assessment, therefore, could have been sufficient in 41.8%, radiology in 44.1%; and endoscopy in 12.2% of the studied population to reach a proper diagnosis. CONCLUSIONS: Clinical assessment is crucial to evaluate chronic nasal obstruction. The lateral nasopharyngeal roentgenogram provides objective evaluation of the adenoid but its limitations should be considered. Nasal endoscopy may substitute for radiology but should be reserved for unusual cases. PMID- 19674360 TI - Clinical characteristics of respiratory syncytial virus infection-associated acute otitis media. AB - BACKGROUND: It is known that children with respiratory syncytial virus (RSV) infection frequently have complications of acute otitis media (AOM). METHODS: The hospital records of 148 inpatients aged 6-35 months who had RSV infection between January 2004 and December 2007, were retrospectively investigated. RESULTS: Forty six out of 148 children (31%) had AOM. There was a significantly greater number of children with fever who had AOM (P = 0.005). The percentage of children with beta-lactamase-non-producing ampicillin-resistant (BLNAR) Haemophilus influenzae in nasopharyngeal culture who had AOM showed a tendency to be greater than that of those who did not have AOM, but this was not statistically significant (P = 0.068). Moreover, BLNAR H. influenzae was positive in middle ear fluid specimens from four of five children with AOM who underwent tympanocentesis. There were no significant differences in the incidence of lower airway infection, leukocytes counts, or serum C-reactive protein levels between children with and without AOM. CONCLUSIONS: Children who had RSV infection with AOM had a higher incidence of fever than those without AOM. PMID- 19674361 TI - Homozygous variant of UGT1A1 gene mutation and severe neonatal hyperbilirubinemia. AB - BACKGROUND: The aim of the present study was to compare, in a case-control study, the prevalence of nucleotide 211 guanine to adenine (G-->A) mutation of uridine diphosphoglucuronosyl transferase (UGT1A1) gene in Malaysian Chinese newborns with and without severe hyperbilirubinemia (total serum bilirubin >250 micromol/L during first 48 h of life or > or =300 micromol/L thereafter), and to determine whether this mutation was a significant risk factor associated with severe hyperbilirubinemia. METHODS: Seventy-four term infants of Chinese descent admitted with severe hyperbilirubinemia were recruited. Infants without severe hyperbilirubinemia (n = 125) were randomly selected from among healthy Chinese term infants. UGT1A1 nucleotide 211 polymorphism was assayed using the Taqman single nucleotide polymorphism genotyping method. Using gestational age, types of feeds, G6PD mutation, G6PD enzyme levels, and UGT1A1 gene mutation status as independent variables, logistic regression analysis was carried out to determine the significant risk factors associated with severe hyperbilirubinemia. RESULTS: UGT1A1 gene mutation was significantly more common among hyperbilirubinemic infants (39.2%) than controls (25.6%; P = 0.04). Gestational age (adjusted odds ratio [OR], 0.7; 95% confidence intervals [CI]: 0.5-0.9; P = 0.01), G6PD mutation (adjusted OR, 7.2; 95%CI: 2.7-19.0; P < 0.0001), exclusive breast-feeding (adjusted OR, 11.7; 95%CI: 2.7-49.9; P = 0.001), and homozygous variant of UGT1A1 gene mutation (adjusted OR, 32.2; 95%CI: 3.8-273.2; P = 0.001) were significant risk factors. Heterozygous variant of UGT1A1 gene mutation, actual levels of G6PD enzyme, and mixed feeding were not. CONCLUSION: Homozygous variant of nucleotide 211 G-->A mutation of UGT1A1 gene is a significant risk factor associated with severe hyperbilirubinemia among Malaysian Chinese newborns. PMID- 19674362 TI - Effects of interview on mood status of pregnant women with high-risk delivery. AB - BACKGROUND: Expectant mothers at high risk for preterm labor admitted to the Mother and Child Health Centers (MCH) often have psychological problems and mood disturbances. The purpose of the present study was to evaluate the effects of an interview by health professionals on their mood status. METHODS: Participants consisted of 52 women admitted to the MCH of Kansai Medical University (KMU) hospital and who had delivered between December 2006 and September 2007. Interview sessions by health professionals consisting of a neonatologist and a clinical psychologist, termed 'KMU baby doctor-team interview', were held once a week. During the interview the neonatologist discussed the physiology of preterm infants and treatment for the possible complications while the clinical psychologist listened to the expectant mothers talk about their anxieties or complaints and responded to their requests as much as possible. To investigate their mood status objectively, the participants were asked to complete the Japanese version of the Profile of Mood States (POMS) before and after the first session. RESULTS: On admission, more than 20% of participants ranging from 19 to 42 years of age had mood disturbances in Tension-Anxiety, Depression-Dejection, and Vigor categorized by POMS. The average score for Depression-Dejection, however, significantly improved after the first interview sessions. CONCLUSIONS: Mood disturbances were observed in a considerable number of expectant mothers at high risk for preterm labor. Interviews by health professionals consisting of a neonatologist and a clinical psychologist may alter their mood status. PMID- 19674363 TI - Reliability of family history of lifestyle-related diseases on questionnaire. AB - BACKGROUND: Family history of lifestyle-related diseases is an important risk factor and is widely used in epidemiologic studies. To justify its use the validity of the family history needs to be assessed. The aim of the present study was therefore to examine reliability, that is, consistency in repeated measurements, of the family history obtained on questionnaire. METHODS: A questionnaire survey on the family history of lifestyle-related diseases was conducted for 438 high school students about students' parents, grandparents, uncles and aunts, twice with a 1 year interval. The questionnaire was filled in by their parents. Discordance between the two questionnaires in age, age at death and a positive history of lifestyle-related diseases expressed by age at onset by decade age among the family members was examined. Kappa was used as a measure of agreement. RESULTS: The kappas for relatives were high, mostly >0.7 for hypertension and >0.5 for myocardial infarction, angina pectoris, stroke, diabetes mellitus and hypercholesterolemia. The discordance tended to be higher among grandparents than parents, uncles and aunts. The discordance with regard to relatives was mostly <4% for angina pectoris, myocardial infarction, stroke and diabetes mellitus; for hypertension and hypercholesterolemia the discordance was <6%. CONCLUSIONS: The questionnaire provides a reliable and valid method of collecting family history on relatives of high school students. Thus, family history obtained through questionnaires can be used justifiably as a risk factor for lifestyle-related diseases among children and youths. PMID- 19674364 TI - Urinary uric acid : creatinine ratios in healthy Turkish children. AB - BACKGROUND: Determining uric acid : creatinine ratios in random urine samples may be useful to assess the excretion of uric acid in children. Because it was shown that urinary uric acid excretion varies with age and geographic area, it is important to have accurate reference values of uric acid excretion. The aim of the present study was therefore to obtain regional reference values for urinary uric acid : creatinine ratios in healthy Turkish children. METHODS: A total of 1306 children aged 1 month-15 years were analyzed for uric acid and creatinine, and urinary uric acid : creatinine ratios were determined from each sample. The second non-fasting morning urine samples were taken from all the children. Urine samples were analyzed for uric acid using the uricase method, and for creatinine with the Jaffe reaction. RESULTS: The mean +/- SD and 5th-95th percentiles of urinary uric acid : creatinine ratios (mg/mg) were 1.09 +/- 0.48 and 0.27-1.87 at 1-6 months, 0.86 +/- 0.41 and 0.19-1.64 at 7-12 months, 0.76 +/- 0.32 and 0.32 1.43 at 1-3 years, 0.63 +/- 0.29 and 0.20-1.23 at 4-6 years, 0.44 +/- 0.24 and 0.14-0.93 at 7-11 years, and 0.30 +/- 0.14 and 0.12-0.62 at 12-15 years. Uric acid : creatinine ratios were not significantly different between boys and the girls except at 12-15 years. Girls aged 12-15 years had higher urinary uric acid : creatinine ratio when compared with boys (P < 0.05). There was no correlation between urinary uric acid : creatinine ratio and protein intake. CONCLUSIONS: Urinary uric acid : creatinine ratio changes with age. When assessing urinary uric acid : creatinine ratio, the clinician should consider the age of the child. PMID- 19674365 TI - Diagnostic determination system for high-risk screening for inborn errors of bile acid metabolism based on an analysis of urinary bile acids using gas chromatography-mass spectrometry: results for 10 years in Japan. AB - BACKGROUND: Some patients with cholestasis of unknown cause may have inborn errors of bile acid metabolism (IEBAM) thus causing abnormalities of bile acid biosynthesis. Although seven types of bile acid synthetic defects have thus far been reported for this disorder, no detailed information on its incidence and so on in Japan is yet available. In order to elucidate the current status of IEBAM in Japan, in July 1996 a diagnostic determination system was established for high risk screening for IEBAM. METHODS: Urinary bile acids were analyzed on gas chromatography-mass spectrometry (GC-MS) and quantitative analysis was done using selected ion monitoring (SIM). RESULTS AND CONCLUSIONS: In a total of 576 samples analyzed over the 10 year period prior to June 2005, 159 patients were found with cholestasis of unknown etiology. Of these patients, 10 (6.3%) were found to have IEBAM by this system, while 91 (61.1%) had cholestasis without a definitive diagnosis. This diagnostic determination system with GC-MS of urinary bile acids is therefore considered useful for detecting IEBAM. PMID- 19674366 TI - Diagnostic quality of Actigraph-based physical activity cut-offs for children: what overweight/obesity references can tell? AB - BACKGROUND: There is conflicting literature about Actigraph-based cut-off points (CoP) that define the lower limit of moderate-to-vigorous physical activity (MVPA) among children. The aim of the present study was to compare the diagnostic quality of Actigraph-based CoP relative to different body mass index (BMI) classification systems of the weight status of children. METHODS: One hundred and thirteen children aged 8-11 years recruited from three randomly selected elementary schools wore the Actigraph, model 7164 (Actigraph Manufacturing Technology) for 3 days. Five CoP, expressed in counts per minute (c.p.m.; CoP 1000 CoP 2000, CoP 3000, CoP 3200, and CoP 3600) were used to compute their MVPA. The area under the receiver operating characteristic curves (AUC) together with specific indices allowed assessment of the performance of these CoP in reference to overweight/obesity status as defined using BMI-based criteria from the International Obesity Task Force (IOTF), French References (FR) and World Health Organization (WHO) standards. RESULTS: Overweight/obesity frequency ranged from 30% to 42%, with no sex-related difference. All AUC (range, 0.623-0.660) were significantly higher than 0.500 except from those of CoP 1000 and 2000 using the IOTF criteria (0.602 +/- 0.058 and 0.601 +/- 0.057, respectively) and the FR (0.608 +/- 0.060 for CoP 1000). Furthermore, with the WHO standards, all the CoP provided non-significant AUC (range, 0.566-0.597). According to the IOTF and the FR criteria, respectively, CoP 3600 had the highest probability of correct decision (0.62 and 0.68), the lowest misclassification errors (0.38 and 0.32), the highest validity coefficient (0.21 and 0.29), and the highest expected maximum utility (59 and 83). CONCLUSION: When children are classified using BMI based criteria, the threshold of 3600 c.p.m. should be more appropriate in discriminating non-overweight from overweight/obese. PMID- 19674367 TI - Effect of growth hormone therapy in two sisters with SHOX haploinsufficiency. PMID- 19674368 TI - Single episode of Behcet's disease-like symptoms caused by herpes simplex virus reactivation. PMID- 19674369 TI - Late-onset, unusual neurological symptoms in children with mycoplasma infection. PMID- 19674370 TI - Male newborn with anti-Kg antibody-induced hemolytic disease of the newborn. PMID- 19674371 TI - Second Japanese case of neonatal alloimmune thrombocytopenia due to anti-human platelet antigen-5a. PMID- 19674372 TI - Early diagnosis for polyarthritis of juvenile idiopathic arthritis using systemic gallium scintigraphy. PMID- 19674373 TI - Early prophylactic thyroidectomy for RET mutation-positive MEN 2B. PMID- 19674374 TI - Rodriguez lethal acrofacial dysostosis syndrome with pulmonary hypoplasia. PMID- 19674375 TI - Juvenile generalized acanthosis nigricans without any systemic disease. PMID- 19674376 TI - Vitamin B6 deficiency presenting with low alanine aminotransferase in a critically ill child. PMID- 19674379 TI - Childhood emotional abuse and dissociation in patients with conversion symptoms. AB - AIM: The aim of the present study was to evaluate the relationship between reported childhood trauma and dissociation in patients who have a conversion symptom. METHOD: Thirty-two outpatients with a conversion symptom were evaluated using Dissociative Experiences Scale, Somatoform Dissociation Questionnaire, Childhood Trauma Questionnaire, Spielberger Trait Anxiety Inventory, Clinician Administered Dissociative State Scale, and Dissociative Disorders Interview Schedule. RESULTS: A DSM-IV dissociative disorder was diagnosed in 46.9% of the patients. Conversion patients with a dissociative disorder had borderline personality disorder more frequently than those without a dissociative disorder. Among childhood trauma types, emotional abuse was the only significant predictor of dissociation in regression analysis. None of the childhood trauma types predicted borderline personality disorder criteria. CONCLUSIONS: Borderline personality disorder, dissociation and reports of childhood emotional abuse refer to a subgroup among patients with conversion symptom. Dissociation seems to be a mediator between childhood trauma and borderline phenomena among these patients. PMID- 19674380 TI - Effect of age and gender on schizotypal personality traits in the normal population. AB - AIM: There is some evidence that age and gender influence expression of schizotypal traits in the normal population. One important limitation of previous studies, however, is the restricted age range of subjects. The aim of the present study was therefore to investigate the effect of gender and age on schizotypal personality features. METHODS: A total of 1024 participants aged between 16 and 90 were assessed using the schizotypal personality questionnaire. The effect of gender and age on schizotypal features was investigated. Confirmatory factor analysis was used to examine the factor structure of the scale in different age groups. RESULTS: Male subjects scored higher than women in negative and disorganized-like symptoms. Female subjects scored significantly in social anxiety and odd belief subscales. Younger participants significantly scored higher in reference, odd belief, unusual perceptual experiences, odd behavior and odd speech subscales. While decreases in schizotypy scores were gradual for most of the scales, increased disorganization in female subjects was restricted to late adolescence. Confirmatory factor analysis of the data supported modified versions of the Raine three-factor model across all age groups. CONCLUSION: Psychological changes related to adolescence and better social adaptation as a result of life experience might contribute to the age- and gender-related differences in schizotypy. PMID- 19674381 TI - Prevalence of i.v. thiopental use in psychiatric emergency settings in Japan. AB - AIM: Because i.v. barbiturates such as thiopental carry the risk of apnea and laryngeal spasm in asthmatic patients, reducing the use of barbiturate in emergency situations is important. The purpose of the present study was therefore to investigate the prevalence of i.v. thiopental as a choice of sedation in behavioral emergency settings, we conducted a cross-sectional multicenter study. METHODS: Psychiatric emergency departments of seven hospitals were studied during a 4-month period. Patients with a score >15 on the Excited Component of the Positive and Negative Syndrome Scale (PANSS-EC) who received i.v. medication were included in the study. Drugs were chosen according to the Japanese guidelines, in which the first injection was either haloperidol or benzodiazepine in accordance with clinical requirements. A second injection, which was the opposite drug to the first injection was administered as needed. Only when excitement obviously increased following the first injection, which was considered uncontrollable without thiopental according to expert experience, was thiopental given as a second injection. A total of 137 patients were included. The mean age was 40.4 years (SD 13.1), and the rate of male gender, drug-naive, and F2 (schizophrenia, schizotypal and delusional disorders) on the ICD-10 were 48.9%, 29.9%, and 65.7%, respectively. RESULTS: The rate of patients treated with thiopental as a second injection was 8.0% (n = 11). All of the first injections in patients treated with thiopental were not haloperidol but benzodiazepines (P = 0.0072). CONCLUSION: Because this multicenter study has an epidemiological character, the prevalence of i.v. thiopental use in psychiatric emergency settings in Japan is considered to be 8.0%. PMID- 19674382 TI - Algorithm-guided treatment versus treatment as usual for major depression. AB - AIMS: The remission rates for patients with major depressive disorder (MDD) during algorithm-guided treatment (AGT), which consisted of four treatment strategy steps were prospectively compared with treatment as usual (TAU). METHODS: The remission rates of patients with mild or moderate MDD during AGT (n = 83) were compared with TAU (n = 127). RESULTS: The remission rate in the AGT group (60.2%) was approximately 10% greater than that in the TAU group (49.7%). The median number of days to achieve remission in the AGT group (93 days) was half as long as that in the TAU group (191 days). The hazard ratio of remission was 1.5 (95% confidence interval: = 1.2-1.8). A higher rate of lithium augmentation in the AGT group (20.5%) compared to the TAU (4.7%) may have led to the greater remission rate. Most participants who did not achieve remission either during the initial or second treatment steps dropped out from AGT. CONCLUSIONS: AGT may be superior to TAU for patients with mild or moderate MDD, based on the remission rates achieved. The later treatment steps in the AGT, however, were rarely utilized because participants who did not receive any benefit dropped out early. PMID- 19674383 TI - Total antioxidant capacity and total oxidant status in patients with major depression: impact of antidepressant treatment. AB - AIM: The purpose of the present study was to investigate whether total antioxidant capacity (TAC) and total oxidant status (TOS) are associated with major depressive disorder (MDD) and to evaluate the impact of antidepressant treatment on TAC and TOS in MDD. METHODS: Fifty-seven MDD patients and 40 healthy controls participated in the study. Serum TAC and TOS were measured both in patients and controls using Erel's methods. Patients were treated with antidepressant drugs for 12 weeks. The treatment course was evaluated using the Montgomery-Asberg Depression Rating Scale (MADRS) in all patients. RESULTS: TOS and oxidative stress index (OSI) were higher (P = 0.0001 for both) and TAC was lower (P = 0.0001) in the MDD group compared with those of the controls. After 3 months of antidepressant treatment, TOS and OSI were decreased and TAC was increased compared with the pretreatment values (P = 0.0001, for all). Furthermore, there were significant positive correlations between the severity of the disease and serum TOS and OSI (r = 0.584, P = 0.0001; r = 0.636, P = 0.0001, respectively). A negative correlation was found between the severity of the disease and serum TAC (r = -0.553, P = 0.0001) at the pre-treatment stage. CONCLUSION: Treatment administered for 3 months to MDD patients increases TAC while decreasing TOS and OSI. PMID- 19674384 TI - Practice-based depression screening for psychiatry outpatients: feasibility comparison of two-types of Center for Epidemiologic Studies Depression Scales. AB - AIMS: The Center for Epidemiologic Studies Depression Scale (CES-D) has been validated to avoid misdiagnoses of major depression in routine psychiatric outpatient settings, but it was reported to be only marginally feasible in these specific settings. A briefer and simpler version, known as the 10-item CES-D, meant to attain adequate feasibility, has been validated in geriatric outpatient settings, but it has not yet been examined in psychiatry outpatient settings. The purpose of the present study was therefore to compare the feasibility, reliability, and validity of the two types of CES-D. METHODS: A cross-sectional analysis was conducted of 86 consecutive outpatients in a psychiatric department in a general hospital. RESULTS: The 10-item CES-D has a higher feasibility than the 20-item CES-D, and its internal consistency, reliability, and validity are almost identical to those of the 20-item CES-D. CONCLUSIONS: The 10-item CES-D is the better instrument to use because of the higher feasibility than the 20-item CES-D in psychiatric outpatient settings. The different answer format used in each questionnaire (a yes or no format in the former vs a multiple-choice format in the latter) may influence the feasibility, rather than the number of items. PMID- 19674385 TI - Atypical antipsychotics in the treatment of delirium. AB - Delirium is common in all medical settings. Atypical antipsychotics are increasingly used for the management of delirium symptomatology but their effectiveness has not been systematically studied. The aim of the present study was therefore to provide an up-to-date review on the use of atypical antipsychotics in the treatment of delirium. A search was conducted of the databases of MEDLINE, PsycINFO and EMBASE from 1997 to 2008 for English-language articles using the key words 'delirium' and the names of all the atypical antipsychotics. A total of 23 studies were used for this review. Fifteen of the studies were single-agent trials. Four studies were comparison trials, including one double-blind trial, and four studies were retrospective, including three comparison studies. All studies reported improvement of delirium symptomatology after the administration of atypical antipsychotics. No study included a placebo group. Other limitations included sample heterogeneity, small sample size, different rating scales for delirium, and lack of adequate controls. The improvement in delirium was observed within a few days after treatment initiation and the doses given were relatively low. Atypical antipsychotics were well tolerated, but safety was not evaluated systematically. Atypical antipsychotics appear to be effective and safe in symptomatic treatment of delirium but the evidence is limited and inconclusive. There are no double-blind, placebo controlled studies assessing the efficacy and safety of these agents in delirium. Further research is needed with well-designed studies. PMID- 19674386 TI - Dishonesty in the name of equity. AB - Under what conditions do people act dishonestly to help or hurt others? We addressed this question by examining the influence of a previously overlooked factor-the beneficiary or victim of dishonest acts. In two experiments, we randomly paired participants and manipulated their wealth levels through an initial lottery. We then observed how inequity between partners influenced the likelihood of one dishonestly helping or hurting the other, while varying the financial incentives for dishonest behavior. The results show that financial self interest cannot fully explain people's tendency to dishonestly help or hurt others. Rather, such dishonesty is influenced by emotional reactions to wealth based inequity, even when the dishonesty bears a personal financial cost. Envy evoked by negative inequity led to hurting behavior, whereas guilt induced by positive inequity motivated helping behavior. Finally, inequity between the partner and third parties triggered dishonest helping through empathy with the partner. PMID- 19674387 TI - Choosing to enter or avoid diagnostic social situations. AB - Three studies suggest that people control the nature of their relationships, in part, by choosing to enter (or avoid) situations providing feedback about other people's social interest. In Study 1, chronically avoidant individuals (but not others) preferred social options that would provide no information about other people's evaluations of them over social options that would, but did not prefer nondiagnostic situations more generally. In Study 2, chronically avoidant students (but not others) in a methods class preferred to have their teacher assign them to working groups (a nondiagnostic situation) over forming their own groups (a diagnostic situation). In Study 3, individuals experimentally primed to feel avoidant were less likely than those primed to feel secure to choose to receive feedback about how another person felt about them. Overall, the research suggests that choices of socially diagnostic versus socially nondiagnostic situations play an important role in guiding people's social relationships. PMID- 19674388 TI - Eighteen-month-old infants show increased helping following priming with affiliation. AB - We show that the mere hint of affiliation dramatically increases prosocial behavior in infants. Eighteen-month-old infants helped a person in need more often, and more spontaneously, when primed with photographs evoking affiliation than when primed with photographs evoking individuality. This study demonstrates that social primes can have an influence on infant behavior, and so opens up a wealth of possibilities for future research. In addition, these data have wide ranging practical implications, suggesting that subtle changes to the social environment can promote prosocial behavior in children. PMID- 19674389 TI - An electrophysiological measure of access to representations in visual working memory. AB - Previous research has demonstrated that the maintenance of visual information in working memory is associated with a sustained posterior contralateral negativity. Here we show that this component is also elicited during the spatially selective access to visual working memory. Participants memorized a bilateral visual search array that contained two potential targets on the left and right side. The task relevant side was signalled by post-cues that were presented either 150 ms after array offset or after a longer interval (700-1000 ms). Enhanced negativities at posterior electrodes contralateral to the cued side of a target were elicited in response to both early and late post-cues, suggesting that they reflect not only memory maintenance, but also processes involved in the access to stored visual working memory representations. Results provide new electrophysiological evidence for the retinotopic organization of visual working memory. PMID- 19674390 TI - Modeling single-trial LRP waveforms using gamma functions. AB - The lateralized readiness potential (LRP) is a component of average event-related potentials that has proven very useful in the study of hand-specific motor preparation. We developed a model of single-trial LRP waveforms that produces realistic average waveforms for both stimulus-locked and response-locked averaging. This model may be useful in computer simulation studies of LRP scoring methods, and it may open up the possibility of ultimately retrieving trial-by trial information about LRP activity. PMID- 19674391 TI - Error-related negativity predicts academic performance. AB - Activity in the anterior cingulate cortex (ACC) has been linked to the processes of error detection and conflict monitoring, along with the subsequent engagement of cognitive-control mechanisms. The error-related negativity (ERN) is an electrophysiological signal associated with this ACC monitoring process, occurring approximately 100 ms after an error is made. The current study examined the possibility that individual differences in ERN magnitude would predict performance outcomes related to cognitive control. Undergraduate students completed a color-naming Stroop task while their neural activity was recorded via electroencephalogram. Results indicated that a larger ERN following errors was significantly correlated with better academic performance as measured by official student transcripts. A greater ability to monitor performance and engage cognitive-control mechanisms when needed thus appears associated with improved real-world performance. PMID- 19674393 TI - ERP correlates of online monitoring of auditory feedback during vocalization. AB - When speakers hear the fundamental frequency (F0) of their voice altered, they shift their F0 in the direction opposite the perturbation. The current study used ERPs to examine sensory processing of short feedback perturbations during an ongoing utterance. In one session, participants produced a vowel at an F0 of their own choosing. In another session, participants matched the F0 of a cue voice. An F0 perturbation of 0, 25, 50, 100, or 200 cents was introduced for 100 ms. A mismatch negativity (MMN) was observed. Differences between sessions were only found for 200-cent perturbations. Reduced compensation when speakers experienced the 200-cent perturbations suggests that this larger perturbation was perceived as externally generated. The presence of an MMN, and no earlier (N100) response suggests that the underlying sensory process used to identify and compensate for errors in mid-utterance may differ from feedback monitoring at utterance onset. PMID- 19674392 TI - Relationship between the P3 event-related potential, its associated time frequency components, and externalizing psychopathology. AB - P3 amplitude reduction (P3-AR) is associated with biological vulnerability to a spectrum of externalizing disorders, such as ADHD, conduct disorder, and substance use disorders. P3, however, is generally characterized as a broad activation involving multiple neurophysiological processes. One approach to separating P3-related processes is time-frequency (TF) analysis. The current study used a novel PCA-based TF analysis method to investigate relationships between P3, its associated TF components, and externalizing in a community-based sample of adolescent males. Results showed that 1) alone, P3 and each TF-PCA derived component could successfully discriminate diagnostic groups from controls, and 2) delta components in specific time ranges accounted for variance beyond that accounted for by P3. One delta component was associated with all diagnostic groups, suggesting it may represent a more parsimonious endophenotype for externalizing than P3-AR. PMID- 19674394 TI - On the differentiation of N2 components in an appetitive choice task: evidence for the revised Reinforcement Sensitivity Theory. AB - Task- and personality-related modulations of the N2 were probed within the framework of the revised Reinforcement Sensitivity Theory (RST). Using an appetitive choice task, we investigated 58 students with extreme scores on the behavioral inhibition system and behavioral approach system (BIS/BAS) scales. The baseline-to-peak N2 amplitude was sensitive to the strength of decision conflict and demonstrated RST-related personality differences. In addition to the baseline N2 amplitude, temporal PCA results suggested two N2 components accounting for a laterality effect and capturing different N2 patterns for BIS/BAS groups with increasing conflict level. Evidence for RST-related personality differences was obtained for baseline-to-peak N2 and tPCA components in the present task. The results support the RST prediction that BAS sensitivity modulates conflict processing and confirm the cognitive-motivational conflict concept of RST. PMID- 19674395 TI - Early life stress and psychiatric disorder modulate cortical responses to affective stimuli. AB - Altered affective processing has been proposed as mediating between early life stress (ELS) and subsequent psychopathology. The present study examined whether ELS influences affective cortical processing differently in psychiatric patients and healthy subjects. The number of stressful experiences before onset of puberty was assessed in 50 inpatients with diagnoses of Major Depressive Disorder, schizophrenia, drug addiction, or Borderline Personality Disorder and in 20 healthy comparison subjects. Subjects monitored pleasant, neutral, and unpleasant pictures during magnetoencephalographic recording. Suppression of right-posterior activity 160-210 ms after stimulus onset was associated with certain diagnoses and high ELS. Results confirmed specific contributions of ELS versus adult stress, comorbid posttraumatic stress disorder, or depression. PMID- 19674396 TI - Effects of prior stimulus and prior perception on neural correlates of auditory stream segregation. AB - We examined whether effects of prior experience are mediated by distinct brain processes from those processing current stimulus features. We recorded event related potentials (ERPs) during an auditory stream segregation task that presented an adaptation sequence with a small, intermediate, or large frequency separation between low and high tones (Deltaf), followed by a test sequence with intermediate Deltaf. Perception of two streams during the test was facilitated by small prior Deltaf and by prior perception of two streams and was accompanied by more positive ERPs. The scalp topography of these perception-related changes in ERPs was different from that observed for ERP modulations due to increasing the current Deltaf. These results reveal complex interactions between stimulus-driven activity and temporal-context-based processes and suggest a complex set of brain areas involved in modulating perception based on current and previous experience. PMID- 19674397 TI - Enhanced long-term recollection for emotional pictures: evidence from high density ERPs. AB - The present study used behavioral and electrophysiological measures to investigate the processes mediating long-term recognition memory for emotional and neutral pictures. The results show enhanced memory recollection for emotional arousing pictures compared to neutral low arousing pictures. In accordance with the behavioral data, we observed enhanced old/new effects in the ERPs for emotionally arousing pictures in the recollection-sensitive old/new component at centro-parietal sites (500-800 ms). Moreover, early old/new effects were present over frontal and parietal sites (300-500 ms) irrespective of picture contents. Analysis of the subjective awareness, indexed by the confidence ratings, showed that the late parietal old/new effect was increased for high confidence responses whereas the early component (300-500 ms) was mainly driven by low confidence responses, an indication for familiarity based recognition processes. PMID- 19674398 TI - The effects of valence and arousal on the neural activity leading to subsequent memory. AB - This study examined how valence and arousal affect the processes linked to subsequent memory for emotional information. While undergoing an fMRI scan, participants viewed neutral pictures and emotional pictures varying by valence and arousal. After the scan, participants performed a recognition test. Subsequent memory for negative or high arousal information was associated with occipital and temporal activity, whereas memory for positive or low arousal information was associated with frontal activity. Regression analyses confirmed that for negative or high arousal items, temporal lobe activity was the strongest predictor of later memory whereas for positive or low arousal items, frontal activity corresponded most strongly with later memory. These results suggest that the types of encoding processes relating to memory (e.g., sensory vs. elaborative processing) can differ based on the affective qualities of emotional information. PMID- 19674399 TI - Working memory resources in young children with mathematical difficulties. AB - Working memory (WM) (Baddeley, 1986, 1997) is argued to be one of the most important cognitive resources underlying mathematical competence (Geary, 2004). Research has established close links between WM deficits and mathematical difficulties. This study investigated the possible deficits in WM, language and fluid intelligence that seem to characterize 4- to 6-year-old children with poor early mathematical skills before formal mathematics education. Children with early mathematical difficulties showed poor performance in both verbal and visuospatial WM tasks as well as on language tests and a fluid intelligence test indicating a thoroughly lower cognitive base. Poor WM performance was not moderated by fluid intelligence, but the extent of WM deficits was related to language skills. The educational implications are discussed. PMID- 19674400 TI - The Stress Strategy Test in relation to personality traits and cognitive abilities. AB - Relations between results of the Stress Strategy Test (SST), a computerized test the first author developed partly involving the Stroop effect, and of an interview and an intelligence test used by the Swedish armed forces as selection instruments for placement of recruits in type of military service were investigated in a group of 3,205 recruits. The SST results were found able to explain 17.64% and 12.25% of the variance in results of the intelligence test and the interview, respectively. Emotional stability and psychological energy were the two interview variables most closely related to results of the SST. The usefulness of the SST in a selection context is discussed. PMID- 19674401 TI - Personality impressions associated with four distinct humor styles. AB - This study examined how personality impressions about another person are influenced by the style of humor that person displays. Four distinct styles were examined, with two of these being adaptive (affiliative and self-enhancing humor), and two being maladaptive (aggressive and self-defeating humor). Participants read descriptions of an individual displaying each humor style, and then rated that individual on several other personality attributes (e.g., friendly, complaining). The adaptive humor styles enhanced personality impressions of another individual, whereas the maladaptive styles had strong detrimental effects. Furthermore, participants provided clearly differentiated personality impressions within both the adaptive and maladaptive humor categories. Affiliative humor led to more positive impressions of another than self-enhancing humor; whereas aggressive humor resulted in more negative personality impressions than self-defeating humor. These findings were discussed in terms of approaches to humor that acknowledge the multifaceted nature of this construct and the resulting impact on social relationships. PMID- 19674402 TI - Adolescents' own suggestions for bullying interventions at age 13 and 16. AB - In this study we examined adolescents' perspectives on what interventions they consider to be effective in order to stop the bullying of a student. The adolescents' suggestions were reviewed at two time points, age 13 and 16. Participants were 474 girls and 403 boys at the first point of examination, and 429 girls and 332 boys at the second point of examination. The participants' suggestions were divided into categories based on some of the anti-bullying strategies commonly presented by researchers. Results showed that some anti bullying strategies were more salient than others in the adolescents' suggestions, and that their suggestions differed as a function of age, sex and to some extent, current experience of victimization. Having serious talks with the students involved was among the most common suggestions at both ages. However, girls were more likely than boys, and non-victims were more likely than victims, to suggest this particular strategy. PMID- 19674403 TI - Cancer, acute stress disorder, and repressive coping. AB - The purpose of this study was to investigate the association between repressive coping style and Acute Stress Disorder (ASD) in a sample of cancer patients. A total of 112 cancer patients recently diagnosed with cancer participated in the study. ASD was assessed by the Stanford Acute Stress Reaction Questionnaire, and repressive coping was assessed by a combination of scores from the Marlowe-Crowne Social Desirability Scale, and the Bendig version of the Taylor Manifest Anxiety Scale. Significantly fewer patients classified as "repressors" were diagnosed with ASD compared to patients classified as "non-repressors". However, further investigations revealed that the lower incidence of ASD in repressors apparently was caused by a low score on anxiety and not by an interaction effect between anxiety and defensiveness. Future studies have to investigate whether different psychological mechanisms are responsible for the lower incidence of ASD in repressors and true low-anxious patients. PMID- 19674404 TI - Creating coherence in real-life decision processes: Reasons, differentiation and consolidation. AB - Differentiation and Consolidation Theory describes human decision making as a process in which attractiveness values are restructured in order to reach a decision and support the decision made. Here, the theory was developed to include reasons pro and con alternatives and tested on students making decisions between two university psychotherapy training programs (cognitive-behavioral and psychodynamic therapy). Before and also after the decision, the attractiveness of the chosen alternative was upgraded and the non-chosen alternative downgraded. Different measures of evaluations of an alternative, such as "best" or "worse" converged over time until shortly after the decision. The number of reasons pro and con alternatives give a more complete picture than attractiveness and increased from the first to the last session. The reasons supporting the chosen alternative increased in strength, but reasons against the non-chosen alternative decreased. In informal comments participants reported that the study also served as a decision aid. PMID- 19674405 TI - The chloroplastic lipocalin AtCHL prevents lipid peroxidation and protects Arabidopsis against oxidative stress. AB - Lipocalins are small ligand-binding proteins with a simple tertiary structure that gives them the ability to bind small, generally hydrophobic, molecules. Recent studies have shown that animal lipocalins play important roles in the regulation of developmental processes and are involved in tolerance to oxidative stress. Plants also possess various types of lipocalins, and bioinformatics analyses have predicted that some lipocalin members may be present in the chloroplast. Here we report the functional characterization of the Arabidopsis thaliana chloroplastic lipocalin AtCHL. Cellular fractionation showed that AtCHL is a thylakoid lumenal protein. Drought, high light, paraquat and abscisic acid treatments induce AtCHL transcript and protein accumulation. Under normal growth conditions, knockout (KO) and over-expressing (OEX) lines do not differ from wild type plants in terms of phenotype and photosynthetic performance. However, KO plants, which do not accumulate AtCHL, show more damage upon photo-oxidative stress induced by drought, high light or paraquat. In contrast, a high level of AtCHL allows OEX plants to cope better with these stress conditions. When exposed to excess light, KO plants display a rapid accumulation of hydroxy fatty acids relative to the wild-type, whereas the lipid peroxidation level remains very low in OEX plants. The increased lipid peroxidation in KO plants is mediated by singlet oxygen and is not correlated with photo-inhibition of the photosystems. This work provides evidence suggesting that AtCHL is involved in the protection of thylakoidal membrane lipids against reactive oxygen species, especially singlet oxygen, produced in excess light. PMID- 19674406 TI - A redox-active isopropylmalate dehydrogenase functions in the biosynthesis of glucosinolates and leucine in Arabidopsis. AB - We report a detailed functional characterization of an Arabidopsis isopropylmalate dehydrogenase (AtIPMDH1) that is involved in both glucosinolate biosynthesis and leucine biosynthesis. AtIPMDH1 shares high homology with enzymes from bacteria and yeast that are known to function in leucine biosynthesis. In plants, AtIPMDH1 is co-expressed with nearly all the genes known to be involved in aliphatic glucosinolate biosynthesis. Mutation of AtIPMDH1 leads to a significant reduction in the levels of free leucine and of glucosinolates with side chains of four or more carbons. Complementation of the mutant phenotype by ectopic expression of AtIPMDH1, together with the enzyme's substrate specificity, implicates AtIPMDH1 in both glucosinolate and leucine biosynthesis. This functional assignment is substantiated by subcellular localization of the protein in the chloroplast stroma, and the gene expression patterns in various tissues. Interestingly, AtIPMDH1 activity is regulated by a thiol-based redox modification. This work characterized an enzyme in plants that catalyzes the oxidative decarboxylation step in both leucine biosynthesis (primary metabolism) and methionine chain elongation of glucosinolates (specialized metabolism). It provides evidence for the hypothesis that the two pathways share a common origin, and suggests a role for redox regulation of glucosinolate and leucine synthesis in plants. PMID- 19674407 TI - Ectopic expression of MYB46 identifies transcriptional regulatory genes involved in secondary wall biosynthesis in Arabidopsis. AB - MYB46 functions as a transcriptional switch that turns on the genes necessary for secondary wall biosynthesis. Elucidating the transcriptional regulatory network immediately downstream of MYB46 is crucial to our understanding of the molecular and biochemical processes involved in the biosynthesis and deposition of secondary walls in plants. To gain insights into MYB46-mediated transcriptional regulation, we first established an inducible secondary wall thickening system in Arabidopsis by expressing MYB46 under the control of dexamethasone-inducible promoter. Then, we used an ATH1 GeneChip microarray and Illumina digital gene expression system to obtain a series of transcriptome profiles with regard to the induction of secondary wall development. These analyses allowed us to identify a group of transcription factors whose expression coincided with or preceded the induction of secondary wall biosynthetic genes. A transient transcriptional activation assay was used to confirm the hierarchical relationships among the transcription factors in the network. The in vivo assay showed that MYB46 transcriptionally activates downstream target transcription factors, three of which (AtC3H14, MYB52 and MYB63) were shown to be able to activate secondary wall biosynthesis genes. AtC3H14 activated the transcription of all of the secondary wall biosynthesis genes tested, suggesting that AtC3H14 may be another master regulator of secondary wall biosynthesis. The transcription factors identified here may include direct activators of secondary wall biosynthesis genes. The present study discovered novel hierarchical relationships among the transcription factors involved in the transcriptional regulation of secondary wall biosynthesis, and generated several testable hypotheses. PMID- 19674409 TI - Influence of canal preparation depth on the incidence of femoral medullary infarction with Zurich Cementless Canine Total Hip arthroplasty. AB - OBJECTIVE: To determine the incidence of femoral medullary infarction after modifying the depth of femoral reaming and filing when performing total hip replacement (THR) using the Zurich Cementless Total Hip Replacement system (ZCTHR). STUDY DESIGN: Case series. ANIMALS: Dogs (n=31) that had ZCTHR (34). METHODS: Thirty-one dogs (34 THR) had ZCTHR (May 2003-September 2006) and with >1 year radiographic (craniocaudal, mediolateral views) follow-up after THR were evaluated for the presence of femoral medullary infarcts. Incidence was compared with a previous study performed before the technique modification. RESULTS: Femoral medullary infarction occurred in 1 femur (2.9%; dog <18 months at THR) compared with 19.5% before the technique change, a significant decrease (P<.001). CONCLUSION: Limiting the depth of reaming and filing of the medullary canal resulted in a significant decrease in the incidence of femoral medullary infarction. CLINICAL RELEVANCE: Depth of reaming and filing the medullary canal should be limited when performing THR using the ZCTHR. PMID- 19674410 TI - Femoral medullary infarction prevalence with the Zurich Cementless Canine Total Hip arthroplasty. AB - OBJECTIVE: To document the prevalence of femoral medullary infarction associated with the Zurich Cementless Total Hip Replacement (ZCTHR) system in dogs. STUDY DESIGN: Case series. ANIMALS: Dogs (n=35) with 41 ZCTHR implants. METHODS: Medical records (February 1999-December 2002) were reviewed for dogs that had ZCTHR and at least 1 year follow-up with radiographic evaluation. Thirty-five dogs (41 ZCTHR) met the inclusion criteria. Femoral morphologic data, implant to bone relationships, and medullary infarcts were recorded. Data were analyzed for associations between infarct occurrence and morphologic details and dog characteristics. RESULTS: Eight of 41 femurs had radiographic evidence of infarcts (19.5%). Dogs with infarcts were significantly younger (mean [+/-SD] age, 18.5+/-5.2 months) compared with those without infarction (mean age, 44.4+/ 5.6 months; P=.027). None of the other variables were significantly different between dogs with and without infarcts. Three femurs with infarcts ultimately developed stem loosening. CONCLUSION: A higher prevalence of femoral medullary infarcts was identified with ZCTHR compared with incidence reported for other total hip systems. Younger dogs were more likely to develop infarction. CLINICAL RELEVANCE: Femoral infarction appears to be associated with stem loosening. Adjustments in surgical technique or delaying surgery beyond 18 months of age may reduce incidence of infarction but needs further evaluation. PMID- 19674411 TI - A lateral approach for screw repair in lag fashion of spiral third metacarpal and metatarsal medial condylar fractures in horses. AB - OBJECTIVE: To describe a lateral approach for screw fixation in lag fashion of simple spiral medial condylar fractures of the third metacarpus/metatarsus (MC3/MT3). STUDY DESIGN: Case series. ANIMALS: Thoroughbred racehorses (n=9). METHODS: Nondisplaced medial MC3/MT3 condylar fractures (3 thoracic, 6 pelvic limbs), with mean length 126 mm (range, 91-151 mm) were repaired by internal fixation, under general anesthesia, using multiple 4.5 mm cortical screws inserted in lag fashion from the lateral aspect of the limb, using radiographic or fluoroscopic guidance. Horses were recovered from anesthesia in half-limb casts; 7 unassisted and 2 using a rope-recovery system. Horses had 2 months box rest, 1 month in-hand walking, and follow-up radiographic examination at 3 months. RESULTS: Horses recovered uneventfully from anesthesia. Five horses raced; 1 returned to training, was persistently lame, and was retired to stud; 2 were retired directly to stud; and 1 horse was lost to follow-up. CONCLUSIONS: MC3/MT3 medial condylar fractures were successfully repaired by screws inserted n lag fashion form the lateral aspect. CLINICAL RELEVANCE: Use of a lateral approach to medial condylar MC3/MT3 fractures allows screw insertion perpendicular to the fracture plane without interference with palmar/plantar soft tissue structures or from the splint bones. Although repair was performed under general anesthesia, the technique should be adaptable to application in standing horses. PMID- 19674412 TI - A lateral approach to the repair of propagating fractures of the medial condyle of the third metacarpal and metatarsal bone in 18 racehorses. AB - OBJECTIVES: To report the technique, observations on fracture configurations and results of treatment by fixation lag screw following the fracture plane determined by an approach to the third metacarpal/metatarsal bone (MC3/MT3) that begins laterally over the metacarpo(metatarso)phalangeal joint and extends dorsally over the diaphysis of the bone. STUDY DESIGN: Case series. ANIMALS: Thoroughbred horses (n=18) with propagating fractures of the medial condyle of MC3/MT3. METHODS: Retrospective analysis of case records of horses with fractures of the medial condyle of MC3/MT3 that propagated sagittaly or in a spiral configuration into the diaphysis, repaired surgically under general anesthesia by screw fixation in lag fashion through a lateral approach with periosteal reflection. RESULTS: Fractures were readily identified at surgery, enabling screw fixation in lag fashion following the fracture plane. Fracture configurations varied and could be classified as sagittal and spiral fractures with fractures within each group generally following a similar course. All horses recovered relatively uneventfully from general anesthesia and surgery, and all fractures healed well. Thirteen horses returned to training; 5 subsequently raced. CONCLUSIONS: Repair of propagating sagittal and spiral fractures of the medial condyle of MC3/MT3 with diaphyseal involvement, through a lateral approach with periosteal reflection permits stable fixation with minimal complications. In this series there were no catastrophic failures. CLINICAL RELEVANCE: Fractures of the medial condyle of MC3/MT3 that propagate either sagittaly or in a spiral configuration into the diaphysis can be successfully repaired with screw fixation in lag fashion using a lateral approach with periosteal reflection. PMID- 19674413 TI - Mapping of split-line pattern and cartilage thickness of selected donor and recipient sites for autologous osteochondral transplantation in the canine stifle joint. AB - OBJECTIVE: To characterize donor and recipient sites for autologous osteochondral transplantation in the canine stifle joint with respect to split-line pattern and cartilage thickness. STUDY DESIGN: In vitro study. Sample Population- Stifle joints (n=30) of dogs >20 kg. METHODS: Collagen network orientation of the hyaline cartilage coverage of the distal femur was assessed using split-line technique (n=10). Cartilage thickness was measured radiographically on osteochondral plugs harvested at the abaxial and axial surfaces of the femoropatellar joint (n=15), the most proximal abaxial aspect of the medial condyle (CO; n=5) and at both femoral condyles (n=15). RESULTS: Cartilage within the femoropatellar joint and both femoral condyles had mostly transversely orientated pattern of split-lines. Abaxial to the femoropatellar joint split lines were absent. All donor sites had significantly thinner cartilage than the medial condyle (CM). Only the distal trochlea had cartilage thickness comparable to the lateral condyle (CL). The thinnest cartilage was found abaxial to the femoropatellar joint followed by CO, which was approximately 2-3 times thinner than the cartilage at both condyles. CONCLUSIONS: None of the investigated donor sites provided transplants of comparable cartilage thickness to CM. Transplants from within the femoropatellar joint should be harvested in priority as they reach cartilage morphology comparable to CL. CLINICAL RELEVANCE: Adjusting for cartilage thickness and split-line pattern may improve long-term outcome after autologous osteochondral transplantation. But surface curvature and donor-site morbidity have to be considered as well when choosing the most suitable donor site. PMID- 19674414 TI - Cervical spine locking plate fixation for treatment of cervical spondylotic myelopathy in large breed dogs. AB - OBJECTIVE: To describe indirect decompression by means of cervical spine locking plate (CSLP) fixation with vertebral distraction, discectomy, and cancellous block bone grafting in large breed dogs with single caudal cervical dynamic spondylotic lesions diagnosed by myelography with linear traction to the cervical spine, and contrast-enhanced computed tomography. STUDY DESIGN: Prospective clinical study. ANIMALS: Dogs (n=12) with caudal cervical spondylotic myelopathy because of a single dynamic, traction-responsive lesion. METHODS: Single, traction-responsive, caudal cervical spondylotic lesions were treated by vertebral distraction, discectomy, cancellous block bone grafting, and CSLP fixation. Follow-up was obtained by sequential recheck examination by the author or referring veterinarian or by telephone inquiries. RESULTS: Ten dogs had neurologic improvement after surgery. Indirect decompression by maintained distraction with cancellous block grafting and CSLP fixation was readily accomplished with less risk of blood loss or iatrogenic spinal cord injury than that associated with direct (ventral) decompression. There were no complications of graft intrusion, extrusion or subsidence, implant loosening, foraminal impingement, or end-plate failure. Two dogs that had satisfactory short-term recoveries developed clinical signs associated with adjacent segment disease and were euthanatized. At long-term follow-up, 8 dogs had satisfactory function, either a normal gait or one with slight to moderate proprioceptive deficits. CONCLUSIONS: CSLP fixation with cancellous block interbody grafting is an effective and perhaps safer method of treating single-level, traction-responsive cervical spondylosis in large breed dogs. CLINICAL RELEVANCE: CSLP fixation with interbody bone grafting is a viable alternative to other techniques for treatment of single-level, traction-responsive cervical spondylosis. PMID- 19674415 TI - An in vitro biomechanical comparison of dynamic condylar screw plate combined with a dorsal plate and double plate fixation of distal diaphyseal radial osteotomies in adult horses. AB - OBJECTIVE: To compare stiffness and strength of a dynamic condylar screw plate combined with dorsal broad dynamic compression plate (DCS-bDCP) fixation with double broad dynamic compression plate (dbDCP) fixation used to repair oblique distal fractures of adult equine radii. STUDY DESIGN: Experimental. SAMPLE POPULATION: Adult equine radii (n=10 pair). METHODS: An unconstrained three dimensional loading-measurement system was used to determine stiffness of a 50 mm long intact, and then DCS-bDCP or dbDCP-plated osteotomized/ostectomized segment of radii when subjected to a nondestructive sequence of compression, torsion, and lateral-to-medial (LM), medial-to-lateral (ML), cranial-to-caudal (CrCa), and caudal-to-cranial (CaCr) bending. Uniform load over the entire length of construct identified its weakest characteristics during torsion and LM and CrCa bending to failure. RESULTS: No difference was observed between osteotomized/ostectomized DCS-bDCP and dbDCP construct stiffness for all 6 loading modes, and strength for all 3 failure loads. Ostectomized DCS-bDCP and dbDCP construct stiffness was significantly lower than osteotomized radii, the latter approaching intact for axial, LM, and CrCa bending. Most frequent failure was bone fracture through exit site of a screw located adjacent to osteotomy/ostectomy. CONCLUSIONS: DCS-DCP and dbDCP constructs had comparable strength and stiffness when repairing osteotomies/ostectomies in equine adult radius bone. Fracture reduction increased stiffness that approached intact bone for loads that placed the unplated side in compression. CLINICAL RELEVANCE: DCS bDCP and dbDCP constructs are comparable in stiffness and strength when applied to oblique distal diaphyseal osteotomies/ostectomies in equine radius bone. However, the DCS's localized effect on distal epiphyseal structure because of additional bone removal remains to be investigated under in vivo articular loading conditions. PMID- 19674416 TI - Efficacy of adhesive incise drapes in preventing bacterial contamination of clean canine surgical wounds. AB - OBJECTIVE: To investigate the efficacy of adhesive incise drapes in reducing bacterial contamination of clean canine surgical wounds. STUDY DESIGN: Randomized clinical trial. ANIMALS: Dogs (n=100) having elective ovariohysterectomy or stifle surgery. METHODS: Dogs were randomly assigned to 1 of 2 groups: drape or no drape. Swabs obtained from the inner edge of the surgical wound at the beginning (swab 1) and end (swab 2) of surgery were submitted for microbial culture. Number of colony forming units was counted for all positive cultures and change in bacterial counts between swabs 1 and 2 was calculated. Percentage adhesive drape adherence at the end of surgery was calculated from a digital photograph of the surgical site. duration of surgery/anesthesia and the anesthetic induction agent used were recorded. RESULTS: There was a significant increase in bacterial counts between swabs 1 and 2 (P=.001). Wound contamination was 14% (6 drape; 8 no drape; P=0.78) with Staphylococcus spp. most commonly isolated. Median percentage drape adherence at the end of surgery was 89.3% (0 100%). Duration of anesthesia was significantly related to wound contamination (P=.013), but duration of surgery and anesthesia induction agent were not. CONCLUSIONS: Adhesive incise drapes did not reduce wound contamination of clean canine surgical wounds. CLINICAL RELEVANCE: Use of adhesive incise drapes in clean surgical procedures is of questionable benefit in dogs. PMID- 19674417 TI - Prospective evaluation of two intracorporeally sutured prophylactic laparoscopic gastropexy techniques compared with laparoscopic-assisted gastropexy in dogs. AB - OBJECTIVE: To report technique, surgical time, complication rate, and postoperative activity in dogs undergoing 2 intracorporeally-sutured total laparoscopic gastropexy (TLG) techniques compared with a laparoscopic-assisted gastropexy (LAG) technique. STUDY DESIGN: Randomized clinical trial. ANIMALS: Dogs (n=30) weighing >25 kg. METHODS: Dogs were randomly assigned to 1 of 3 groups. Two TLG techniques were performed using a median 3 portal technique. One of 2 suturing techniques was used to approximate corresponding incisions made in the stomach and body wall; using intracorporeal hand-suturing or a suture-assist device (Endostitch). In a 3rd group, a previously reported LAG technique was used. All dogs had an activity monitor placed for 7 days pre- and postoperatively. Linear regression analyses were performed to evaluate the association of surgical procedure on gastropexy time and the percentage change in activity counts. RESULTS: Median gastropexy time was 28 minutes (range, 20-41 minutes) for LAG, 48 minutes (range, 39-61 minutes) for the hand suture TLG technique, and 56 minutes (range, 30-90 minutes) for the Endostitch TLG technique. LAG was performed faster than TLG (P<.05). LAG dogs had a greater decrease in postoperative activity than TLG dogs (P=.005); however there was no difference in surgical time or postoperative activity between TLG techniques. CONCLUSIONS: TLG can be performed safely and effectively in dogs and although it takes longer, it has less impact on postoperative activity compared with LAG. CLINICAL RELEVANCE: TLG techniques may have advantages over LAG as measured by a greater willingness of dogs to move around postoperatively. PMID- 19674418 TI - Long-term efficacy of a percutaneously adjustable hydraulic urethral sphincter for treatment of urinary incontinence in four dogs. AB - OBJECTIVE: To evaluate the efficacy of a surgically placed, static hydraulic urethral sphincter (SHUS) for treatment of urethral sphincter mechanism incompetency (USMI). STUDY DESIGN: Prospective study. ANIMALS: Spayed female dogs (n=4) with acquired USMI. METHODS: Urinary incontinence was assessed using a subjective continence score before and after implantation of an SHUS on the proximal urethra via ventral median celiotomy. Dogs were assessed for urinary continence, urinary tract infections, and implant-associated complications for 30 months. Residual incontinence was treated with percutaneous inflation of the SHUS with sterile saline solution through a biocompatible subcutaneous administration port. RESULTS: At last follow-up (26-30 months after surgery), continence scores improved from a median preoperative score of 3/10 to a median postoperative score of 10. One dog developed wound drainage over the subcutaneously placed administration port but remained continent after port removal. Three occluders were percutaneously filled with additional saline (median, 0.18 mL; mean, 0.16 mL) to improve continence after surgery. CONCLUSIONS: Application and adjustment of an SHUS provided sustained improvements in continence score in all dogs. CLINICAL RELEVANCE: In this pilot study, 3 of 4 dogs with hydraulic urethral sphincter implantation had successful percutaneous adjustment and maintained improved continence scores for 2 years after surgery. Continence was maintained in the 4th dog even after administration port removal. Based on this pilot study, the SHUS warrants further clinical evaluation for treatment of dogs with USMI unresponsive to medical management. PMID- 19674419 TI - Correlation of surface electromyography of the vastus lateralis muscle in dogs at a walk with joint kinematics and ground reaction forces. AB - OBJECTIVE: To describe the activity pattern of the vastus lateralis muscle in dogs at walk measured by surface electromyography (EMG) in relation to kinematics and kinetics of the pelvic limb. STUDY DESIGN: Experimental. ANIMALS: Malinois dogs (n=11). METHODS: Dogs (mean +/- SD age, 5.5 +/- 2.9 years; weight, 27.3 +/- 3.8 kg; shoulder height, 62.7 +/- 3.3 cm) walked on a treadmill system with integrated force plates, which allowed simultaneous analysis of kinematics, kinetics, and EMG data from all limbs. The maxima, minima, and their time of occurrence in the motion cycle of the EMG and the pelvic limb kinematics and kinetics were calculated; correlations between joint movement patterns, ground reaction forces (GRF), and activity pattern of the muscle group were investigated. RESULTS: The vastus lateralis muscle had an activity pattern with 2 peaks and a close positive correlation with GRF. The 1st peak occurred in early stance, followed by a decrease in activity during mid-stance. The 2nd peak occurred directly before the quick activity decrease in late stance phase, reaching its minimum early in swing phase. CONCLUSIONS: These results suggest that the vastus lateralis muscle supports the vertical position and elevation of the pelvis during stance and push-off. During early stance, the muscle acts as a coantagonist to the hamstring muscle group and the gastrocnemius muscle, and restrains flexion during the late stance. CLINICAL RELEVANCE: Results of this study could enhance diagnosis of locomotor system disorders and facilitate monitoring effects of treatments (e.g., therapeutic exercises) on gait ability and muscle function. PMID- 19674420 TI - Acute phase response to surgery of varying intensity in horses: a preliminary study. AB - OBJECTIVE: To evaluate the postoperative inflammatory response of horses to elective surgery of varying intensity. STUDY DESIGN: Prospective longitudinal study. ANIMALS: Horses referred to 2 hospitals for either arthroscopic removal of a unilateral osteochondritic lesion in the tibiotarsal joint (minimal surgical trauma, n=11), correction of recurrent laryngeal neuropathy by laryngoplasty and ventriculectomy (intermediate surgical trauma, n=10) or removal of an ovarian tumor by laparotomy (major surgical trauma, n=5). METHODS: Horses had a thorough clinical examination every day. White blood cell (WBC) counts and concentrations of serum amyloid A (SAA), fibrinogen, and iron were assessed in blood samples obtained before, and 1-3, 5, 7, 9, and 11 days after surgery. Differences in levels of the inflammatory markers between the 3 surgical groups were analyzed using repeated measures ANOVA. RESULTS: Postoperative concentrations of SAA and fibrinogen were significantly higher in horses that had laparotomy and ovariectomy than in horses that had laryngoplasty and ventriculectomy, or arthroscopy. Iron concentrations decreased to lower levels after intermediate and major surgical trauma than after small surgical trauma. WBC count did not differ between the 3 groups. CONCLUSIONS: Levels of SAA, fibrinogen, and iron reflected the intensity of the surgical trauma, whereas WBC count did not. CLINICAL RELEVANCE: Postoperative measurements of SAA, fibrinogen, and iron may be useful for comparing surgical trauma associated with new and established surgical techniques. Moreover, knowledge of the normal postoperative acute phase response is essential, if acute phase reactants are to be used for monitoring occurrence of postoperative infections. PMID- 19674421 TI - Achilles tendon repair in dogs using the semitendinosus muscle: surgical technique and short-term outcome in five dogs. AB - OBJECTIVE: To describe augmentation of primary Achilles tendon repair using suture with a semitendinosus muscle flap and report outcome in 5 dogs. STUDY DESIGN: Prospective clinical study. ANIMALS: Dogs (n=5) with Achilles tendon rupture (n=6). METHODS: After tendon repair with #2 polypropylene in a 3-loop pulley suture pattern, the lateral one-half of the semitendinosus muscle was transected from the ischium, rotated distally then sutured with #2 polypropylene to the calcaneus in a 3-loop pulley pattern. The epitenon was sutured to the muscle flap fascia with interrupted sutures. All dogs had a bivalved cast for 2 weeks then a cranial splint for 2-6 weeks. Lameness scores (0=stands and walks normally to 4=non-weight-bearing lameness, plantigrade stance on affected pelvic limb) were determined every 2-3 weeks postoperatively for 12 weeks. Outcome was determined from telephone questionnaire of owners. RESULTS: Four had lameness scores of 0, the 5th had a score of 1 at 12 weeks. Three owners were very satisfied with outcome. Minor complications included cast sores (2 dogs), infection (2), and acute swelling (1); 1 major complication occurred (infection resulting in reoperation). CONCLUSIONS: Semitendinosus flap augmentation resulted in early return to function without prolonged postoperative immobilization. Three dogs returned to full work/activity after repair. CLINICAL RELEVANCE: Augmentation of primary Achilles tendon repair with a semitendinosus flap can be considered in dogs with chronic rupture but further investigation of the long term outcome using this technique is needed. PMID- 19674422 TI - Immunohistochemical localization of RANK, RANKL and OPG in healthy and arthritic canine elbow joints. AB - OBJECTIVE: To determine if the receptor activator of nuclear factor-kappaB receptor activator of nuclear factor-kappaB ligand-osteoprotegerin (RANK-RANKL OPG) system is active in bone remodeling in dogs and, if so, whether differences in expression of these mediators occur in healthy and arthritic joints. STUDY DESIGN: Experimental study. SAMPLE POPULATION: Fragmented processus coronoidei (n=20) were surgically removed from dogs with elbow arthritis and 5 corresponding healthy samples from dogs euthanatized for reasons other than elbow joint disease. METHODS: Bright-field immunohistochemistry and high-resolution fluorescence microscopy were used to investigate the distribution of RANK, RANKL, and OPG in healthy and arthritic joints. RESULTS: All 3 molecules were identified by immunostaining of canine bone tissue. In elbow dysplasia, the number of RANK positive osteoclasts was increased. In their vicinity, cells expressing RANKL, a mediator of osteoclast activation, were abundant whereas the number of osteoblasts having the potential to limit osteoclastogenesis and bone resorption via OPG was few. CONCLUSIONS: The RANK-RANKL-OPG system is active in bone remodeling in dogs. In elbow dysplasia, a surplus of molecules promoting osteoclastogenesis was evident and is indicative of an imbalance between the mediators regulating bone resorption and bone formation. Both OPG and neutralizing antibodies against RANKL have the potential to counterbalance bone resorption. CLINICAL RELEVANCE: Therapeutic use of neutralizing antibodies against RANKL to inhibit osteoclast activation warrants further investigation. PMID- 19674423 TI - Multiple rib fracture repair in a neonatal Holstein calf. AB - OBJECTIVES: To report rib fracture repair in a neonatal calf using the Securos Cranial Cruciate Ligament Repair System (SCCLRS). STUDY DESIGN: Case report. ANIMALS: A 2-day-old female Holstein calf with fracture of right ribs 4-10. METHODS: On the day of admission the calf was anesthetized and rib fractures were repaired using open reduction and the SCCLRS. RESULTS: Rib fractures were successfully stabilized and the calf discharged from the hospital 8 days postoperatively. CONCLUSIONS: Modification of previously reported use of the SCCLRS to repair rib fractures in foals was required because of the different anatomy in the calf. This new technique was rapidly and easily performed for a large number of fractured ribs in this case. CLINICAL RELEVANCE: Effective rib fracture repair in calves can be readily performed using the SCCLRS with modification of the technique reported in foals. PMID- 19674424 TI - Filling the gaps between performance incentive programs and health care quality improvement. PMID- 19674425 TI - Commentary: A systematic review of health care efficiency measures. PMID- 19674426 TI - Commentary: Slack resources in health care organizations--fat to be trimmed or muscle to be exercised? PMID- 19674427 TI - Effects of the Premier Hospital Quality Incentive Demonstration on Medicare patient mortality and cost. AB - OBJECTIVE: To evaluate the effects of the Premier Inc. and Centers for Medicare and Medicaid Services Hospital Quality Incentive Demonstration (PHQID), a public quality reporting and pay-for-performance (P4P) program, on Medicare patient mortality, cost, and outlier classification. DATA SOURCES: The 2000-2006 Medicare inpatient claims, Medicare denominator files, and Medicare Provider of Service files. STUDY DESIGN: Panel data econometric methods are applied to a retrospective cohort of 11,232,452 admissions from 6,713,928 patients with principal diagnoses of acute myocardial infarction (AMI), heart failure, pneumonia, or a coronary-artery bypass grafting (CABG) procedure from 3,570 acute care hospitals between 2000 and 2006. Three estimators are used to evaluate the effects of the PHQID on risk-adjusted (RA) mortality, cost, and outlier classification in the presence of unobserved selection, resulting from the PHQID being voluntary: fixed effects (FE), FE estimated in the subset of hospitals eligible for the PHQID, and difference-in-difference-in-differences. DATA EXTRACTION METHODS: Data were obtained from CMS. Principal Findings. This analysis found no evidence that the PHQID had a significant effect on RA 30-day mortality or RA 60-day cost for AMI, heart failure, pneumonia, or CABG and weak evidence that the PHQID increased RA outlier classification for heart failure and pneumonia. CONCLUSIONS: By not reducing mortality or cost growth, this study suggests that the PHQID has made little impact on the value of inpatient care purchased by Medicare. PMID- 19674428 TI - The relationship between hospital volume and mortality in mechanical ventilation: an instrumental variable analysis. AB - OBJECTIVE: To examine the relationship between hospital volume and mortality for nonsurgical patients receiving mechanical ventilation. DATA SOURCES: Pennsylvania state discharge records from July 1, 2004, to June 30, 2006, linked to the Pennsylvania Department of Health death records and the 2000 United States Census. STUDY DESIGN: We categorized all general acute care hospitals in Pennsylvania (n=169) by the annual number of nonsurgical, mechanically ventilated discharges according to previous criteria. To estimate the relationship between annual volume and 30-day mortality, we fit linear probability models using administrative risk adjustment, clinical risk adjustment, and an instrumental variable approach. PRINCIPLE FINDINGS: Using a clinical measure of risk adjustment, we observed a significant reduction in the probability of 30-day mortality at higher volume hospitals (>or=300 admissions per year) compared with lower volume hospitals (<300 patients per year; absolute risk reduction: 3.4%, p=.04). No significant volume-outcome relationship was observed using only administrative risk adjustment. Using the distance from the patient's home to the nearest higher volume hospital as an instrument, the volume-outcome relationship was greater than observed using clinical risk adjustment (absolute risk reduction: 7.0%, p=.01). CONCLUSIONS: Care in higher volume hospitals is independently associated with a reduction in mortality for patients receiving mechanical ventilation. Adequate risk adjustment is essential in order to obtained unbiased estimates of the volume-outcome relationship. PMID- 19674430 TI - The health effects of Medicare for the near-elderly uninsured. AB - OBJECTIVE: To determine whether Medicare enrollment at age 65 has an effect on the health trajectory of the near-elderly uninsured. DATA SOURCES: Eight biennial waves (1992-2006) of the Health and Retirement Study, a nationally representative panel survey of noninstitutionalized 51-61 year olds and their spouses. STUDY DESIGN: We use a quasi-experimental approach to compare the health effects of insurance for the near-elderly uninsured with previously insured contemporaneous controls. The primary outcome measure is overall self-reported health status combined with mortality (i.e., excellent to very good, good, fair to poor, dead). RESULTS: The change in the trajectory of overall health status for the previously uninsured that can be attributed to Medicare is small and not statistically significant. For every 100 persons in the previously uninsured group, joining Medicare is associated with 0.6 fewer in excellent or very good health (95 percent CI: -4.8, 3.3), 0.3 more in good health (95 percent CI: -3.8, 4.1), 2.5 fewer in fair or poor health (95 percent CI: -7.4, 2.3), and 2.8 more dead (-4.0, 10.0) by age 73. The health trajectory patterns from physician objective health measures are similarly small and not statistically significant. CONCLUSIONS: Medicare coverage at age 65 for the previously uninsured is not linked to improvements in overall health status. PMID- 19674429 TI - Organizational and market influences on physician performance on patient experience measures. AB - OBJECTIVE: To examine the extent to which medical group and market factors are related to individual primary care physician (PCP) performance on patient experience measures. DATA SOURCES: This study employs Clinician and Group CAHPS survey data (n=105,663) from 2,099 adult PCPs belonging to 34 diverse medical groups across California. Medical group directors were interviewed to assess the magnitude and nature of financial incentives directed at individual physicians and the adoption of patient experience improvement strategies. Primary care services area (PCSA) data were used to characterize the market environment of physician practices. STUDY DESIGN: We used multilevel models to estimate the relationship between medical group and market factors and physician performance on each Clinician and Group CAHPS measure. Models statistically controlled for respondent characteristics and accounted for the clustering of respondents within physicians, physicians within medical groups, and medical groups within PCSAs using random effects. PRINCIPAL FINDINGS: Compared with physicians belonging to independent practice associations, physicians belonging to integrated medical groups had better performance on the communication ( p=.007) and care coordination ( p=.03) measures. Physicians belonging to medical groups with greater numbers of PCPs had better performance on all measures. The use of patient experience improvement strategies was not associated with performance. Greater emphasis on productivity and efficiency criteria in individual physician financial incentive formulae was associated with worse access to care ( p=.04). Physicians located in PCSAs with higher area-level deprivation had worse performance on the access to care ( p=.04) and care coordination ( p<.001) measures. CONCLUSIONS: Physicians from integrated medical groups and groups with greater numbers of PCPs performed better on several patient experience measures, suggesting that organized care processes adopted by these groups may enhance patients' experiences. Physicians practicing in markets with high concentrations of vulnerable populations may be disadvantaged by constraints that affect performance. Future studies should clarify the extent to which performance deficits associated with area-level deprivation are modifiable. PMID- 19674431 TI - The within-year concentration of medical care: implications for family out-of pocket expenditure burdens. AB - OBJECTIVE: To examine the within-year concentration of family health care and the resulting exposure of families to short periods of high expenditure burdens. DATA SOURCE: Household data from the pooled 2003 and 2004 Medical Expenditure Panel Survey (MEPS) yielding nationally representative estimates for the nonelderly civilian noninstitutionalized population. STUDY DESIGN: The paper examines the within-year concentration of family medical care use and the frequency with which family out-of-pocket expenditures exceeded 20 percent of family income, computed at the annual, quarterly, and monthly levels. PRINCIPAL FINDINGS: On average among families with medical care, 49 percent of all (charge-weighted) care occurred in a single month, and 63 percent occurred in a single quarter). Nationally, 27 percent of the study population experienced at least 1 month in which out-of-pocket expenditures exceeded 20 percent of income. Monthly 20 percent burden rates were highest among the poor, at 43 percent, and were close to or above 30 percent for all but the highest income group (families above four times the federal poverty line). CONCLUSIONS: Within-year spikes in health care utilization can create financial pressures missed by conventional annual burden analyses. Within-year health-related financial pressures may be especially acute among lower-income families due to low asset holdings. PMID- 19674432 TI - Health care utilization and costs associated with physical and nonphysical-only intimate partner violence. AB - OBJECTIVE: To estimate health care utilization and costs associated with the type of intimate partner violence (IPV) women experience by the timing of their abuse. METHODS: A total of 3,333 women (ages 18-64) were randomly sampled from the membership files of a large health plan located in a metropolitan area and participated in a telephone survey to assess IPV history, including the type of IPV (physical IPV or nonphysical abuse only) and the timing of the abuse (ongoing; recent, not ongoing but occurring in the past 5 years; remote, ending at least 5 years prior). Automated annual health care utilization and costs were assembled over 7.4 years for women with physical IPV and nonphysical abuse only by the time period during which their abuse occurred (ongoing, recent, remote), and compared with those of never-abused women (reference group). RESULTS: Mental health utilization was significantly higher for women with physical or nonphysical abuse only compared with never-abused women-with the highest use among women with ongoing abuse (relative risk for those with ongoing abuse: physical, 2.61; nonphysical, 2.18). Physically abused women also used more emergency department, hospital outpatient, primary care, pharmacy, and specialty services; for emergency department, pharmacy, and specialty care, utilization was the highest for women with ongoing abuse. Total annual health care costs were higher for physically abused women, with the highest costs for ongoing abuse (42 percent higher compared with nonabused women), followed by recent (24 percent higher) and remote abuse (19 percent higher). Women with recent nonphysical abuse only had annual costs that were 33 percent higher than nonabused women. CONCLUSION: Physical and nonphysical abuse contributed to higher health care utilization, particularly mental health services utilization. PMID- 19674434 TI - Factors associated with increasing nursing home closures. AB - PURPOSE: We determine the rate of nursing home closures for 7 years (1999-2005) and examine internal (e.g., quality), organizational (e.g., chain membership), and external (e.g., competition) factors associated with these closures. DESIGN AND METHOD: The names of the closed facilities and dates of closure from state regulators in all 50 states were obtained. This information was linked to the Online Survey, Certification, and Reporting data, which contains information on internal, organizational, and market factors for almost all nursing homes in the United States. RESULTS: One thousand seven hundred and eighty-nine facilities closed over this time period (1999-2005). The average annual rate of closure was about 2 percent of facilities, but the rate of closure was found to be increasing. Nursing homes with higher rates of deficiency citations, hospital based facilities, chain members, small bed size, and facilities located in markets with high levels of competition were more likely to close. High Medicaid occupancy rates were associated with a high likelihood of closure, especially for facilities with low Medicaid reimbursement rates. IMPLICATIONS: As states actively debate about how to redistribute long-term care services/dollars, our findings show that they should be cognizant of the potential these decisions have for facilitating nursing home closures. PMID- 19674433 TI - Preferences, beliefs, and self-management of diabetes. AB - OBJECTIVE: To assess relationships between self-assessed control over life events, subjective beliefs about longevity, time and risk preference, and other factors on use of recommended care for diabetes mellitus (DM), self-assessed control of diabetes, general health, and laboratory measures of HbA1c levels. DATA SOURCES: Health and Retirement Study (HRS) and 2003 HRS Diabetes Study (HRS DS). STUDY DESIGN: We used logit and ordered logit analyses to assess use of recommended care, and subjective and objective measures of health outcomes. DATA COLLECTION: Secondary analysis of HRS and HRS-DS data. PRINCIPAL FINDINGS: Individuals with higher self-assessed control over life events and higher subjective probabilities of living 10 years engaged in more recommended DM care practices and had better self-assessed DM control and general health. However, these beliefs did not influence HbA1c levels. More highly educated and cognitively able persons were more likely to follow care recommendations. There were differences by race/ethnicity in health outcomes, but not in health investment among Hispanics. CONCLUSIONS: Individuals' beliefs about control over life events and longevity influenced health investment and subjective health outcomes, although these beliefs did not translate into differences in HbA1c levels. Hispanics may realize lower returns on health investments, at least for diabetes care. PMID- 19674435 TI - Alzheimer's disease therapeutic research: the path forward. AB - The field of Alzheimer's disease therapeutic research seems poised to bring to clinic the next generation of treatments, moving beyond symptomatic benefits to modification of the underlying neurobiology of the disease. But a series of recent trials has had disappointingly negative results that raise questions about our drug development strategies. Consideration of ongoing programs demonstrates difficult pitfalls. But a clear path forward is emerging. Successful strategies will utilize newly available tools to reconsider issues of diagnosis, assessment and analysis, facilitating the study of new treatments at early stages in the disease process at which they are most likely to yield major clinical benefits. PMID- 19674436 TI - Angiotensins and Alzheimer's disease: a bench to bedside overview. AB - The pathology of Alzheimer's disease (AD) features amyloid beta peptide deposition, intracellular neurofibrillary tangles and deficits in the cholinergic pathway. Abnormal blood pressure is recognised as a risk factor for the development of AD, although the underlying mechanisms remain unproven. This review proposes angiotensins and associated enzymatic pathways as important mediators of recognised but undefined links between blood pressure and AD. Evidence in support of this involvement translates consistently from the most basic in vitro, in vivo and ex vivo experimental paradigms to more complex human based observational and experimental studies, which also fortunately offer potential for therapeutic interventions against AD. PMID- 19674437 TI - Beyond mild cognitive impairment: vascular cognitive impairment, no dementia (VCIND). AB - Identifying the causes of dementia is important in the search for effective preventative and treatment strategies. The concept of mild cognitive impairment (MCI), as prodromal dementia, has been useful but remains controversial since in population-based studies it appears to be a limited predictor of progression to dementia. Recognising the relative contribution of neurodegenerative and vascular causes, as well as their interrelationship, may enhance predictive accuracy. The concept of vascular cognitive impairment (VCI) has been introduced to describe the spectrum of cognitive change related to vascular causes from early cognitive decline to dementia. A recent review of this concept highlighted the need for diagnostic criteria that encompass the full range of the VCI construct. However, very little is known regarding the mildest stage of VCI, generally termed 'vascular cognitive impairment, no dementia' (VCIND). Whether mild cognitive change in the context of neurodegenerative pathologies is distinct from that in the context of cerebrovascular diseases is not known. This is key to the definition of VCIND and whether it is possible to identify this state. Distinguishing between vascular (that is, VCIND) and non-vascular (that is, MCI) cognitive disorders and determining how well each might predict dementia may not be possible due to the overlap in pathologies observed in the older population. Here, we review the concept of VCIND in an effort to identify recent developments and areas of controversy in nosology and the application of VCIND for screening individuals at increased risk of dementia secondary to vascular disease and its risk factors. PMID- 19674438 TI - The human immunodeficiency virus type 1 Vpr protein and its carboxy-terminally truncated form induce apoptosis in tumor cells. AB - The human immunodeficiency virus type 1 (HIV-1) accessory protein Vpr induces apoptosis after cell cycle arrest at the G2 phase in primate cells. We have reported previously that C81, a carboxy-terminally truncated form of Vpr, interferes with cell proliferation and results in apoptosis without G2 arrest. Here, we investigated whether this property of Vpr and C81 could be exploited for use as a potential anticancer agent. First, we demonstrated that C81 induced G1 arrest and apoptosis in all tumor cells tested. In contrast, Vpr resulted in G2 arrest and apoptosis in HeLa and 293 T cells. Vpr also suppressed the damaged-DNA specific binding protein 1 (DDB1) in HepG2 cells, thereby inducing apoptosis without G2 arrest. G2 arrest was restored when DDB1 was overexpressed in cells that also expressed Vpr. Surprisingly, C81 induced G2 arrest when DDB1 was overexpressed in HepG2 cells, but not in HeLa or 293 T cells. Thus, the induction of Vpr- and C81-mediated cell cycle arrest appears to depend on the cell type, whereas apoptosis was observed in all tumor cells tested. Overall, Vpr and C81 have potential as novel therapeutic agents for treatment of cancer. PMID- 19674439 TI - Achilles tendon suture deteriorates tendon capillary blood flow with sustained tissue oxygen saturation - an animal study. AB - BACKGROUND: Treatment of ruptured Achilles tendons currently constitutes of conservative early functional treatment or surgical treatment either by open or minimal invasive techniques. We hypothesize that an experimental Achilles tendon suture in an animal model significantly deteriorates Achilles tendon microcirculation immediately following suturing. METHODS: Fifteen Achilles tendons of eight male Wistar rats (275-325 g) were included. After preparation of the Achilles tendon with a medial paratendinous approach, Achilles tendon microcirculation was assessed using combined Laser-Doppler and spectrophotometry (Oxygen-to-see) regarding:- tendinous capillary blood flow [arbitrary units AU]- tendinous tissue oxygen saturation [%]- tendinous venous filling pressure [rAU]The main body of the Achilles tendon was measured in the center of the suture with 50 Hz. 10 minutes after Achilles tendon suture (6-0 Prolene), a second assessment of microcirculatory parameters was performed. RESULTS: Achilles tendon capillary blood flow decreased by 57% following the suture (70 +/- 30 AU vs. 31 +/- 16 AU; p < 0.001). Tendinous tissue oxygen saturation remained at the same level before and after suture (78 +/- 17% vs. 77 +/- 22%; p = 0.904). Tendinous venous filling pressure increased by 33% (54 +/- 16 AU vs. 72 +/- 20 AU; p = 0.019) after suture. CONCLUSION: Achilles tendon suture in anaesthetised rats causes an acute loss of capillary perfusion and increases postcapillary venous filling pressures indicating venous stasis. The primary hypothesis of this study was confirmed. In contrast, tendinous tissue oxygen saturation remains unchanged excluding acute intratendinous hypoxia within the first 10 minutes after suture. Further changes of oxygen saturation remain unclear. Furthermore, it remains to be determined to what extent reduced capillary blood flow as well as increased postcapillary stasis might influence tendon healing from a microcirculatory point of view in this animal setting. PMID- 19674440 TI - Why don't physicians adhere to guideline recommendations in practice? An analysis of barriers among Dutch general practitioners. AB - BACKGROUND: Despite wide distribution and promotion of clinical practice guidelines, adherence among Dutch general practitioners (GPs) is not optimal. To improve adherence to guidelines, an analysis of barriers to implementation is advocated. Because different recommendations within a guideline can have different barriers, in this study we focus on key recommendations rather than guidelines as a whole, and explore the barriers to implementation perceived by Dutch GPs. METHODS: A qualitative study using six focus groups was conducted, in which 30 GPs participated, with an average of seven per session. Fifty-six key recommendations were derived from twelve national guidelines. In each focus group, barriers to the implementation of the key recommendations of two clinical practice guidelines were discussed. Focus group discussions were audiotaped and transcribed verbatim. Data was analysed by using an existing framework of barriers. RESULTS: The barriers varied largely within guidelines, with each key recommendation having a unique pattern of barriers. The most perceived barriers were lack of agreement with the recommendations due to lack of applicability or lack of evidence (68% of key recommendations), environmental factors such as organisational constraints (52%), lack of knowledge regarding the guideline recommendations (46%), and guideline factors such as unclear or ambiguous guideline recommendations (43%). CONCLUSION: Our study findings suggest a broad range of barriers. As the barriers largely differ within guidelines, tailored and barrier-driven implementation strategies focusing on key recommendations are needed to improve adherence in practice. In addition, guidelines should be more transparent concerning the underlying evidence and applicability, and further efforts are needed to address complex issues such as comorbidity in guidelines. Finally, it might be useful to include focus groups in continuing medical education as an innovative medium for guideline education and implementation. PMID- 19674442 TI - 'ONE HEALTH' and parasitology. PMID- 19674441 TI - CAPIH: a Web interface for comparative analyses and visualization of host-HIV protein-protein interactions. AB - BACKGROUND: The Human Immunodeficiency Virus type one (HIV-1) is the major causing pathogen of the Acquired Immune Deficiency Syndrome (AIDS). A large number of HIV-1-related studies are based on three non-human model animals: chimpanzee, rhesus macaque, and mouse. However, the differences in host-HIV-1 interactions between human and these model organisms have remained unexplored. DESCRIPTION: Here we present CAPIH (Comparative Analysis of Protein Interactions for HIV-1), the first web-based interface to provide comparative information between human and the three model organisms in the context of host-HIV-1 protein interactions. CAPIH identifies genetic changes that occur in HIV-1-interacting host proteins. In a total of 1,370 orthologous protein sets, CAPIH identifies approximately 86,000 amino acid substitutions, approximately 21,000 insertions/deletions, and approximately 33,000 potential post-translational modifications that occur only in one of the four compared species. CAPIH also provides an interactive interface to display the host-HIV-1 protein interaction networks, the presence/absence of orthologous proteins in the model organisms in the networks, the genetic changes that occur in the protein nodes, and the functional domains and potential protein interaction hot sites that may be affected by the genetic changes. The CAPIH interface is freely accessible at http://bioinfo-dbb.nhri.org.tw/capih. CONCLUSION: CAPIH exemplifies that large divergences exist in disease-associated proteins between human and the model animals. Since all of the newly developed medications must be tested in model animals before entering clinical trials, it is advisable that comparative analyses be performed to ensure proper translations of animal-based studies. In the case of AIDS, the host-HIV-1 protein interactions apparently have differed to a great extent among the compared species. An integrated protein network comparison among the four species will probably shed new lights on AIDS studies. PMID- 19674443 TI - Evaluation of changes in postnatal care using the "Parents' Postnatal Sense of Security" instrument and an assessment of the instrument's reliability and validity. AB - BACKGROUND: A sense of security is important for experiences of parenthood in the early postpartum period. The objectives of this study were to evaluate two models of postnatal care using a questionnaire incorporating the Parents' Postpartum Sense of Security (PPSS) instrument and to test the validity of the PPSS instrument. METHODS: Postal surveys were sent to 234 mothers who had experienced two different forms of postnatal care (study group and control group) and returned by 86.8%. These two groups of mothers were compared for total scores on the PPSS instrument. Demographic variables and mothers' opinions about care interventions were also compared and these variables were tested for correlations with the total PPSS score. A regression analysis was carried out to assess areas of midwifery care which might affect a sense of security. The internal consistency and concurrent validity of the instrument were tested for the total population. RESULTS: there were no significant differences between the groups for scores on the PPSS instrument. A total of three variables predicted 26% of the variability on the PPSS scores for the study group and five variables predicted 37% of the variability in the control group. One variable was common to both: "The midwives on the postnatal ward paid attention to the mother as an individual". There were significant correlations between the total PPSS scores and scores for postpartum talks and visits to the breastfeeding clinic. There was also a significant correlation between the single question: "I felt secure during the first postpartum week" and the total PPSS score. Tests for internal consistency and concurrent validity were satisfactory. CONCLUSION: The proposed new model of care neither improved nor impaired mothers' feelings of security the week following birth. Being seen as an individual by the midwife who provides postnatal care may be an important variable for mothers' sense of postnatal security. It is possible that postpartum talks may encourage the processing of childbirth experiences in a positive direction. Availability of breastfeeding support may also add to a sense of security postpartum. The PPSS instrument has shown acceptable reliability and validity. PMID- 19674444 TI - Supervised learning for the automated transcription of spacer classification from spoligotype films. AB - BACKGROUND: Molecular genotyping of bacteria has revolutionized the study of tuberculosis epidemiology, yet these established laboratory techniques typically require subjective and laborious interpretation by trained professionals. In the context of a Tuberculosis Case Contact study in The Gambia we used a reverse hybridization laboratory assay called spoligotype analysis. To facilitate processing of spoligotype images we have developed tools and algorithms to automate the classification and transcription of these data directly to a database while allowing for manual editing. RESULTS: Features extracted from each of the 1849 spots on a spoligo film were classified using two supervised learning algorithms. A graphical user interface allows manual editing of the classification, before export to a database. The application was tested on ten films of differing quality and the results of the best classifier were compared to expert manual classification, giving a median correct classification rate of 98.1% (inter quartile range: 97.1% to 99.2%), with an automated processing time of less than 1 minute per film. CONCLUSION: The software implementation offers considerable time savings over manual processing whilst allowing expert editing of the automated classification. The automatic upload of the classification to a database reduces the chances of transcription errors. PMID- 19674445 TI - Prioritising neonatal medicines research: UK Medicines for Children Research Network scoping survey. AB - BACKGROUND: The dosing regimen and indications for many medicines in current use in neonatology are not well defined. There is a need to prioritise research in this area, but currently there is little information about which drugs are used in UK neonatal units and the research needs in this area as perceived by UK neonatologists. METHODS: The Neonatal Clinical Studies Group (CSG) of the Medicines for Children Research Network (MCRN) undertook a 2 week prospective scoping survey study to establish which medicines are used in UK neonatal units; how many babies are receiving them; and what clinicians (and other health professionals) believe are important issues for future research. RESULTS: 49 out of 116 units responded to at least one element of the survey (42%). 37 units reported the number of neonates who received medicines over a 2 week period. A total of 3924 medicine-patient pairs were reported with 119 different medicines. 70% of medicine-patient pairs involved medicines that were missing either a license or dose for either term or preterm neonates. 4.3% of medicine-patient pairs involved medicines that were missing both license and dose for any neonate. The most common therapeutic gap in need of additional research identified by UK neonatologists was chronic lung disease (21 responding units), followed by patent ductus arteriosus and vitamin supplements (11 responding units for both) CONCLUSION: The research agenda for neonatal medicines can be informed by knowledge of current medicine use and the collective views of the neonatal community. PMID- 19674447 TI - Persistence of colicinogenic Escherichia coli in the mouse gastrointestinal tract. AB - BACKGROUND: The ability of a bacterial strain to competitively exclude or displace other strains can be attributed to the production of narrow spectrum antimicrobials, the bacteriocins. In an attempt to evaluate the importance of bacteriocin production for Escherichia coli strain residence in the gastrointestinal tract, a murine model experimental evolution study was undertaken. RESULTS: Six colicin-producing, yet otherwise isogenic, E. coli strains were administered and established in the large intestine of streptomycin treated mice. The strains' persistence, population density, and doubling time were monitored over a period of 112 days. Early in the experiment only minor differences in population density between the various colicin-producing and the non-producing control strains were detected. However, over time, the density of the control strains plummeted, while that of the colicin-producing strains remained significantly higher (F(7,66) = 2.317; P < 0.0008). CONCLUSION: The data presented here support prior claims that bacteriocin production may play a significant role in the colonization of E. coli in the gastrointestinal tract. Further, this study suggests that the ability to produce bacteriocins may prove to be a critical factor in determining the success of establishing probiotic E. coli in the gastrointestinal tract of humans and animals. PMID- 19674446 TI - Fluorodeoxyglucose-positron emission tomography in the differential diagnosis of early-onset dementia: a prospective, community-based study. AB - BACKGROUND: The aim of this study was to evaluate the diagnostic accuracy of positron emission tomography (PET) using F18 fluorodeoxyglucose (FDG) in the differential diagnosis of early-onset Alzheimer's disease (AD) and other dementias in a community-dwelling population. METHODS: A prospective sample of 102 individuals presenting consecutively to a primary care centre for examination of suspected early-onset dementing diseases. The mean age of symptom onset of dementia in our patients was 60.06 +/- 4.28 years (mean +/- 1 SD, 95% lower confidence intervals (CI) 54.75, upper 63.37). Patients were evaluated using standard clinical criteria for the diagnosis of dementia. Functional neuroimaging data was obtained and nuclear medicine physicians blind to the clinical diagnosis generated FDG-PET diagnoses. Final clinical diagnoses based on all available data were then established and compared against PET diagnoses. RESULTS: Forty-nine patients received a final clinical diagnosis of early-stage AD (MMSE score 20.97 +/- 5.10). There were 29 non-AD demented patients, 11 depressed patients and a miscellaneous group of 13 patients. Among patients with AD, the sensitivity and specificity of FDG-PET was 78% (95% CI: 66-90%) and 81% (95% CI: 68-86%), respectively. The positive likelihood ratio (PLR) for a FDG-PET scan positive for the diagnosis of AD was 4.11 (95% CI: 2.29-7.32) and negative likelihood ratio (NLR) for a negative FDG-PET scan in the absence of AD was 0.27 (95% CI: 0.16 0.46). The pre-test probability was 48% and post-test probability was 79.02%. The specificity of FDG-PET in the differential diagnosis of other dementias, including frontotemporal dementia, was greater than 95%. Recruitment methods in this study provide a sample that may be more representative of patients in the general population and indicate that FDG-PET imaging can contribute to the diagnosis of AD in younger adults with major increases in the positive likelihood rates and post-test probability. CONCLUSION: The high specificity of FDG-PET suggests this technique might help in the diagnosis of frontotemporal dementia and other forms of early-onset dementia. PMID- 19674448 TI - The Bipolar Interactive Psychoeducation (BIPED) study: trial design and protocol. AB - BACKGROUND: Bipolar disorders affect between 3-5% of the population and are associated with considerable lifelong impairment. Since much of the morbidity associated with bipolar disorder is caused by recurrent depressive symptoms, which are often only poorly responsive to antidepressants, there is a need to develop alternative, non-pharmacological interventions. Psychoeducational interventions have emerged as promising long-term therapeutic options for bipolar disorder. METHODS/DESIGN: The study is an exploratory, individually randomised controlled trial. The intervention known as 'Beating Bipolar' is a psychoeducational programme which is delivered via a novel web-based system. We will recruit 100 patients with a diagnosis of DSM-IV bipolar disorder (including type I and type II) currently in clinical remission. The primary outcome is quality of life. This will be compared for those patients who have participated in the psychoeducational programme with those who received treatment as usual. Quality of life will be assessed immediately following the intervention as well as 10 months after randomisation. Secondary outcomes include current depressive and manic symptoms, number of episodes of depression and mania/hypomania experienced during the follow-up period, global functioning, functional impairment and insight. An assessment of costs and a process evaluation will also be conducted which will explore the feasibility and acceptability of the intervention as well as potential barriers to effectiveness. DISCUSSION: Bipolar disorder is common, under-recognised and often poorly managed. It is a chronic, life-long, relapsing condition which has an enormous impact on the individual and the economy. This trial will be the first to explore the effectiveness of a novel web-based psychoeducational intervention for patients with bipolar disorder which has potential to be easily rolled out to patients. TRIAL REGISTRATION: Current Controlled Trials ISRCTN81375447. PMID- 19674449 TI - Acupuncture in acute herpes zoster pain therapy (ACUZoster) - design and protocol of a randomised controlled trial. AB - BACKGROUND: Acute herpes zoster is a prevalent condition. One of its major symptoms is pain, which can highly influence patient's quality of life. Pain therapy is limited. Acupuncture is supposed to soften neuropathic pain conditions and might therefore act as a therapeutic alternative. Objective of the present study is to investigate whether a 4 week semi-standardised acupuncture is non inferior to sham laser acupuncture and the anticonvulsive drug gabapentine in the treatment of pain associated with herpes zoster. METHODS/DESIGN: Three-armed, randomised, placebo-controlled trial with a total follow-up time of 6 months. Up to estimated 336 patients (interim analyses) with acute herpes zoster pain (VAS > 30 mm) will be randomised to one of three groups (a) semi-standardised acupuncture (168 patients); (b) gabapentine with individualised dosage between 900-3600 mg/d (84 patients); (c) sham laser acupuncture. Intervention takes place over 4 weeks, all patients will receive analgesic therapy (non-opioid analgesics: metamizol or paracetamol and opioids: tramadol or morphine). Therapy phase includes 4 weeks in which group (a) and (c) consist of 12 sessions per patient, (b) visits depend on patients needs. Main outcome measure is to assess the alteration of pain intensity before and 1 week after treatment sessions (visual analogue scale VAS 0-100 mm). Secondary outcome measure are: alteration of pain intensity and frequency of pain attacks; alteration of different aspects of pain evaluated by standardised pain questionnaires (NPI, PDI, SES); effects on quality of life (SF 36); analgesic demand; alteration of sensoric perception by systematic quantitative sensory testing (QST); incidence of postherpetic neuralgia; side effects and cost effectiveness. Credibility of treatments will be assessed. DISCUSSION: This study is the first large-scale randomised placebo controlled trial to evaluate the efficacy of acupuncture compared to gabapentine and sham treatment and will provide valuable new information about the clinical and physiological effects of acupuncture and gabapentine in the treatment of acute herpes zoster pain. The study has been pragmatically designed to ensure that the study findings can be implemented into clinical practice if acupuncture can be shown to be an effective treatment strategy in acute herpes zoster pain. TRIAL REGISTRATION: NCT00885586. PMID- 19674450 TI - Increased chemokine signaling in a model of HIV1-associated peripheral neuropathy. AB - Painful distal sensory polyneuropathy (DSP) is the most common neurological complication of HIV1 infection. Although infection with the virus itself is associated with an incidence of DSP, patients are more likely to become symptomatic following initiation of nucleoside reverse transcriptase inhibitor (NRTI) treatment. The chemokines monocyte chemoattractant protein-1 (MCP1/CCL2) and stromal derived factor-1 (SDF1/CXCL12) and their respective receptors, CCR2 and CXCR4, have been implicated in HIV1 related neuropathic pain mechanisms including NRTI treatment in rodents. Utilizing a rodent model that incorporates the viral coat protein, gp120, and the NRTI, 2'3'-dideoxycytidine (ddC), we examined the degree to which chemokine receptor signaling via CCR2 and CXCR4 potentially influences the resultant chronic hypernociceptive behavior. We observed that following unilateral gp120 sciatic nerve administration, rats developed profound tactile hypernociception in the hindpaw ipsilateral to gp120 treatment. Behavioral changes were also present in the hindpaw contralateral to the injury, albeit delayed and less robust. Using immunohistochemical studies, we demonstrated that MCP1 and CCR2 were upregulated by primary sensory neurons in lumbar ganglia by post-operative day (POD) 14. The functional nature of these observations was confirmed using calcium imaging in acutely dissociated lumbar dorsal root ganglion (DRG) derived from gp120 injured rats at POD 14. Tactile hypernociception in gp120 treated animals was reversed following treatment with a CCR2 receptor antagonist at POD 14. Some groups of animals were subjected to gp120 sciatic nerve injury in combination with an injection of ddC at POD 14. This injury paradigm produced pronounced bilateral tactile hypernociception from POD 14-48. More importantly, functional MCP1/CCR2 and SDF1/CXCR4 signaling was present in sensory neurons. In contrast to gp120 treatment alone, the hypernociceptive behavior associated with the injury plus drug combination was only effectively reversed using the CXCR4 antagonist AMD3100. These studies indicate that the functional upregulation of CCR2 and CXCR4 signaling systems following a combination of gp120 and an NRTI are likely to be of central importance to associated DSP and may serve as potential therapeutic targets for treatment of this syndrome. PMID- 19674451 TI - Cardiac CT and MRI guide surgery in impending left ventricular rupture after acute myocardial infarction. AB - We report the case of a 67 year-old patient who presented with worsening chest pain and shortness of breath, four days post acute myocardial infarction. Contrast enhanced computed tomography of the chest ruled out a pulmonary embolus but revealed an unexpected small subepicardial aneurysm (SEA) in the lateral left ventricular wall which was confirmed on cardiac magnetic resonance imaging. Intraoperative palpation of the left lateral wall was guided by the cardiac MRI and CT findings and confirmed the presence of focally thinned and weakened myocardium, covered by epicardial fat. An aneurysmorrhaphy was subsequently performed in addition to coronary bypass surgery and a mitral valve repair. The patient was discharged home on post operative day eight in good condition and is feeling well 2 years after surgery. PMID- 19674452 TI - An integrated review of "unplanned" dialysis initiation: reframing the terminology to "suboptimal" initiation. AB - BACKGROUND: Ideally, care prior to the initiation of dialysis should increase the likelihood that patients start electively outside of the hospital setting with a mature arteriovenous fistula (AVF) or peritoneal dialysis (PD) catheter. However, unplanned dialysis continues to occur in patients both known and unknown to nephrology services, and in both late and early referrals. The objective of this article is to review the clinical and socioeconomic outcomes of unplanned dialysis initiation. The secondary objective is to explore the potential cost implications of reducing the rate of unplanned first dialysis in Canada. METHODS: MEDLINE and EMBASE from inception to 2008 were used to identify studies examining the clinical, economic or quality of life (QoL) outcomes in patients with an unplanned versus planned first dialysis. Data were described in a qualitative manner. RESULTS: Eight European studies (5,805 patients) were reviewed. Duration of hospitalization and mortality was higher for the unplanned versus planned population. Patients undergoing a first unplanned dialysis had significantly worse laboratory parameters and QoL. Rates of unplanned dialysis ranged from 2449%. The total annual burden to the Canadian healthcare system of unplanned dialysis in 2005 was estimated at $33 million in direct hospital costs alone. Reducing the rate of unplanned dialysis by one-half yielded savings ranging from $13.3 to $16.1 million. CONCLUSION: The clinical and socioeconomic impact of unplanned dialysis is significant. To more consistently characterize the unplanned population, the term suboptimal initiation is proposed to include dialysis initiation in hospital and/or with a central venous catheter and/or with a patient not starting on their chronic modality of choice. Further research and implementation of initiatives to reduce the rate of suboptimal initiation of dialysis in Canada are needed. PMID- 19674453 TI - Application of massive parallel sequencing to whole genome SNP discovery in the porcine genome. AB - BACKGROUND: Although the Illumina 1 G Genome Analyzer generates billions of base pairs of sequence data, challenges arise in sequence selection due to the varying sequence quality. Therefore, in the framework of the International Porcine SNP Chip Consortium, this pilot study aimed to evaluate the impact of the quality level of the sequenced bases on mapping quality and identification of true SNPs on a large scale. RESULTS: DNA pooled from five animals from a commercial boar line was digested with DraI; 150-250-bp fragments were isolated and end-sequenced using the Illumina 1 G Genome Analyzer, yielding 70,348,064 sequences 36-bp long. Rules were developed to select sequences, which were then aligned to unique positions in a reference genome. Sequences were selected based on quality, and three thresholds of sequence quality (SQ) were compared. The highest threshold of SQ allowed identification of a larger number of SNPs (17,489), distributed widely across the pig genome. In total, 3,142 SNPs were validated with a success rate of 96%. The correlation between estimated minor allele frequency (MAF) and genotyped MAF was moderate, and SNPs were highly polymorphic in other pig breeds. Lowering the SQ threshold and maintaining the same criteria for SNP identification resulted in the discovery of fewer SNPs (16,768), of which 259 were not identified using higher SQ levels. Validation of SNPs found exclusively in the lower SQ threshold had a success rate of 94% and a low correlation between estimated MAF and genotyped MAF. Base change analysis suggested that the rate of transitions in the pig genome is likely to be similar to that observed in humans. Chromosome X showed reduced nucleotide diversity relative to autosomes, as observed for other species. CONCLUSION: Large numbers of SNPs can be identified reliably by creating strict rules for sequence selection, which simultaneously decreases sequence ambiguity. Selection of sequences using a higher SQ threshold leads to more reliable identification of SNPs. Lower SQ thresholds can be used to guarantee sufficient sequence coverage, resulting in high success rate but less reliable MAF estimation. Nucleotide diversity varies between porcine chromosomes, with the X chromosome showing less variation as observed in other species. PMID- 19674454 TI - Attitudes towards 12-step groups and referral practices in a 12-step naive treatment culture; a survey of addiction professionals in Norway. AB - BACKGROUND: Addressing substance use disorders effectively requires a long-term approach. Substance abuse treatment is typically of short duration; referring patients to Twelve Step based self-help groups (TSGs) - e.g. Narcotics Anonymous, represents a promising complementary recovery resource. Clinicians' attitudes and referral practices towards the TSGs have mainly been studied in countries with high integration of the 12-step philosophy in their substance abuse services and where the TSGs are widely available, such as the US. In Norway, there are currently 294 weekly TSG meetings (6 per 100,000 inhabitants). This study describes clinicians' attitudes and referral practices to TSGs in Norway where health authorities seek to promote self-help participation, but where the treatment culture is unfamiliar with 12-step fellowships. METHODS: Data collected by a self-administered questionnaire, adapted from established US and UK instruments. Information covered the attitudes, knowledge and referral practices towards TSGs among addiction treatment professionals in Norway in mid 2008. RESULTS: The return rate was 79.7% (n = 291). Participants had moderately positive attitude scores towards TSGs, but referral to these groups among Norwegian addiction professionals was low, as was the level of knowledge about TSGs. More than six out of ten did not refer any patients to TSGs in the previous week. Local variation with more referrals to TSGs in the county with the one established 12-step treatment facility was observed. Respondents' integration of the 12-steps in their own treatment work, higher self-efficacy for making a successful referral, and greater TSG knowledge were associated with referring patients. CONCLUSION: Low referral rates to TSGs point to the need for education and training to raise the awareness and knowledge about it among addiction professionals unfamiliar with these 12-step fellowships. Training should focus on the usefulness of these groups for all types of treatment models regardless of therapeutic orientation. Increased knowledge is expected to lead to higher referral rates, which in turn would maximize the likelihood of positive long-term patient outcomes. PMID- 19674455 TI - Potential for a global dynamic of Influenza A (H1N1). AB - BACKGROUND: Geographical and temporal diffusion patterns of a human pandemic due to Swine Origin Influenza Virus (S-OIV) remain uncertain. The extent to which national and international pandemic preparedness plans and control strategies can slow or stop the process is not known. However, despite preparedness efforts, it appears that, particularly in the USA, Mexico, Canada and the UK, local chains of virus transmission can sustain autonomous dynamics which may lead to the next pandemic. Forecasts of influenza experts usually rely on information related to new circulating strains. METHODS: We attempted to quantify the possible spread of the pandemic across a network of 52 major cities and to predict the effect of vaccination against the pandemic strain, if available. Predictions are based on simulations from a stochastic SEIR model. Parameters used in the simulations are set to values consistent with recent estimations from the outbreak in Mexico. RESULTS: We show that a two-wave pandemic dynamic may be observed in Southern hemisphere because of seasonal constraints for a maximum value of the basic reproductive number (R0, max) within a city equal to 1.5 and a mean generation interval (GI) of 2 days. In this case and in the absence of vaccination, attack rates may reach 46% when considering a completely susceptible population. More severe scenarios characterized by higher values of R0, max (2.2) and GI (3.1) yield an attack rate of 77%. By extrapolation, we find that mass vaccination in all countries (i.e. up to 50% of the population) implemented 6 months after the start of the pandemic may reduce the cumulative number of cases by 91% in the case of the low transmissible strain (R0, max = 1.5). This relative reduction is only 44% for R0, max = 2.2 since most of the cases occur in the first 6 months and so before the vaccination campaign. CONCLUSION: Although uncertainties remain about the epidemiological and clinical characteristics of the new influenza strain, this study provides the first analysis of the potential spread of the pandemic and first assessment of the impact of different immunization strategies. PMID- 19674457 TI - Quality of life and persisting symptoms in intensive care unit survivors: implications for care after discharge. AB - BACKGROUND: We assessed the quality of life of ICU survivors using SF-36 at 4 months after ICU discharge and investigated any correlation of PCS and MCS with age, illness severity and hospital or ICU length of stay. We examined the relationship between these variables, persisting physical and psychological symptoms and the perceived benefit of individual patients of follow-up. FINDINGS: For one year, adult patients admitted for multiple organ or advanced respiratory support for greater than 48 hours to a 16-bedded teaching hospital general intensive care unit were identified. Those surviving to discharge were sent a questionnaire at 4 months following ICU discharge assessing quality of life and persisting symptoms. Demographic, length of stay and illness severity data were recorded. Higher or lower scores were divided at the median value. A two-tailed Students t-test assuming equal variances was used for normally-distributed data and Mann-Whitney tests for non-parametric data.87 of 175 questionnaires were returned (50%), but only 65 had sufficient data giving a final response rate of 37%. Elderly patients had increased MCS as compared with younger patients. The PCS was inversely related to hospital LOS. There was a significant correlation between the presence of psychological and physical symptoms and desire for follow up. CONCLUSION: Younger age and prolonged hospital stay are associated with lower mental or physical quality of life and may be targets for rehabilitation. Patients with persisting symptoms at 4 months view follow-up as beneficial and a simple screening questionnaire may identify those likely to attend outpatient services. PMID- 19674456 TI - The cyclin-dependent kinase inhibitor 5, 6-dichloro-1-beta-D ribofuranosylbenzimidazole induces nongenotoxic, DNA replication-independent apoptosis of normal and leukemic cells, regardless of their p53 status. AB - BACKGROUND: Current chemotherapy of human cancers focuses on the DNA damage pathway to induce a p53-mediated cellular response leading to either G1 arrest or apoptosis. However, genotoxic treatments may induce mutations and translocations that result in secondary malignancies or recurrent disease. In addition, about 50% of human cancers are associated with mutations in the p53 gene. Nongenotoxic activation of apoptosis by targeting specific molecular pathways thus provides an attractive therapeutic approach. METHODS: Normal and leukemic cells were evaluated for their sensitivity to 5, 6-dichloro-1-beta-D ribofuranosylbenzimidazole (DRB) through cell viability and caspase activation tests. The apoptotic pathway induced by DRB was analysed by immunfluorescence and immunoblot analysis. H2AX phosphorylation and cell cycle analysis were performed to study the dependance of apoptosis on DNA damage and DNA replication, respectively. To investigate the role of p53 in DRB-induced apoptosis, specific p53 inhibitors were used. Statistical analysis on cell survival was performed with the test of independence. RESULTS: Here we report that DRB, an inhibitor of the transcriptional cyclin-dependent kinases (CDKs) 7 and 9, triggers DNA replication-independent apoptosis in normal and leukemic human cells regardless of their p53 status and without inducing DNA damage. Our data indicate that (i) in p53-competent cells, apoptosis induced by DRB relies on a cytosolic accumulation of p53 and subsequent Bax activation, (ii) in the absence of p53, it may rely on p73, and (iii) it is independent of ATM and NBS1 proteins. Notably, even apoptosis-resistant leukemic cells such as Raji were sensitive to DRB. CONCLUSION: Our results indicate that DRB represents a potentially useful cancer chemotherapeutic strategy that employs both the p53-dependent and -independent apoptotic pathways without inducing genotoxic stress, thereby decreasing the risk of secondary malignancies. PMID- 19674459 TI - The South London and Maudsley NHS Foundation Trust Biomedical Research Centre (SLAM BRC) case register: development and descriptive data. AB - BACKGROUND: Case registers have been used extensively in mental health research. Recent developments in electronic medical records, and in computer software to search and analyse these in anonymised format, have the potential to revolutionise this research tool. METHODS: We describe the development of the South London and Maudsley NHS Foundation Trust (SLAM) Biomedical Research Centre (BRC) Case Register Interactive Search tool (CRIS) which allows research accessible datasets to be derived from SLAM, the largest provider of secondary mental healthcare in Europe. All clinical data, including free text, are available for analysis in the form of anonymised datasets. Development involved both the building of the system and setting in place the necessary security (with both functional and procedural elements). RESULTS: Descriptive data are presented for the Register database as of October 2008. The database at that point included 122,440 cases, 35,396 of whom were receiving active case management under the Care Programme Approach. In terms of gender and ethnicity, the database was reasonably representative of the source population. The most common assigned primary diagnoses were within the ICD mood disorders (n = 12,756) category followed by schizophrenia and related disorders (8158), substance misuse (7749), neuroses (7105) and organic disorders (6414). CONCLUSION: The SLAM BRC Case Register represents a 'new generation' of this research design, built on a long running system of fully electronic clinical records and allowing in-depth secondary analysis of both numerical, string and free text data, whilst preserving anonymity through technical and procedural safeguards. PMID- 19674461 TI - Adolescent idiopathic scoliosis (AIS) treated with arthrodesis and posterior titanium instrumentation: 8 to 12 years follow up without late infection. AB - BACKGROUND: There are no data in the peer-reviewed literature regarding long term results in patients treated for AIS with a posterior titanium instrumentation. Therefore we assessed the outcome in 50 patients treated by titanium implant. METHODS: A total of 50 patients with a mean age of 16.6 years were treated. In all patients, titanium hooks and pedicle screws were used in combination. The demographic data and the pre- and post-operative radiographs of all 50 patients were re-examined, and 49 of the 50 patients (98%) attended a radiological and clinical follow up-examination on average 10.1 years post-operatively. The clinical results were recorded by means of the SRS 24 questionnaire. RESULTS: In the frontal plane, the mean pre-operative thoracic and lumbar curve had been 62.4 degrees and 43.5 degrees respectively, post-operatively the curves were reduced to 26.9 degrees and 16.3 degrees , resulting in a correction rate of 56.9% for thoracic and 62.5% for lumbar curve. At the follow up-evaluation, the Cobb angle of the thoracic and lumbar curve was 31.0 degrees and 21.3 degrees respectively, giving a final correction rate of 50.3% for thoracic, and 51.0% for lumbar curve. 7 of the 50 patients (14.3%) had undergo revision surgery for complications, but complete implant removal was necessary in only one case. Analysis of the SRS 24 questionnaire showed an average score of 95.8 points. CONCLUSION: Posterior titanium instrumentation is a safe and effective procedure in the surgical correction of AIS. In this retrospective study with small patient number, it shows favourable long-term results; in particular, the loss of correction is low, no late infection occurred and there was a very high survival rate of the implant itself. PMID- 19674460 TI - A single amino acid change (Y318F) in the L-arabitol dehydrogenase (LadA) from Aspergillus niger results in a significant increase in affinity for D-sorbitol. AB - BACKGROUND: L-arabitol dehydrogenase (LAD) and xylitol dehydrogenase (XDH) are involved in the degradation of L-arabinose and D-xylose, which are among the most abundant monosaccharides on earth. Previous data demonstrated that LAD and XDH not only differ in the activity on their biological substrate, but also that only XDH has significant activity on D-sorbitol and may therefore be more closely related to D-sorbitol dehydrogenases (SDH). In this study we aimed to identify residues involved in the difference in substrate specificity. RESULTS: Phylogenetic analysis demonstrated that LAD, XDH and SDH form 3 distinct groups of the family of dehydrogenases containing an Alcohol dehydrogenase GroES-like domain (pfam08240) and likely have evolved from a common ancestor. Modelling of LadA and XdhA of the saprobic fungus Aspergillus niger on human SDH identified two residues in LadA (M70 and Y318), that may explain the absence of activity on D-sorbitol. While introduction of the mutation M70F in LadA of A. niger resulted in a nearly complete enzyme inactivation, the Y318F resulted in increased activity for L-arabitol and xylitol. Moreover, the affinity for D-sorbitol was increased in this mutant. CONCLUSION: These data demonstrates that Y318 of LadA contributes significantly to the substrate specificity difference between LAD and XDH/SDH. PMID- 19674458 TI - The utilization of humanized mouse models for the study of human retroviral infections. AB - The development of novel techniques and systems to study human infectious diseases in both an in vitro and in vivo settings is always in high demand. Ideally, small animal models are the most efficient method of studying human afflictions. This is especially evident in the study of the human retroviruses, HIV-1 and HTLV-1, in that current simian animal models, though robust, are often expensive and difficult to maintain. Over the past two decades, the construction of humanized animal models through the transplantation and engraftment of human tissues or progenitor cells into immunocompromised mouse strains has allowed for the development of a reconstituted human tissue scaffold in a small animal system. The utilization of small animal models for retroviral studies required expansion of the early CB-17 scid/scid mouse resulting in animals demonstrating improved engraftment efficiency and infectivity. The implantation of uneducated human immune cells and associated tissue provided the basis for the SCID-hu Thy/Liv and hu-PBL-SCID models. Engraftment efficiency of these tissues was further improved through the integration of the non-obese diabetic (NOD) mutation leading to the creation of NODSCID, NOD/Shi-scid IL2rgamma-/-, and NOD/SCID beta2 microglobulinnull animals. Further efforts at minimizing the response of the innate murine immune system produced the Rag2-/-gammac-/- model which marked an important advancement in the use of human CD34+ hematopoietic stem cells. Together, these animal models have revolutionized the investigation of retroviral infections in vivo. PMID- 19674462 TI - Current experiences and educational preferences of general practitioners and staff caring for people with dementia living in residential facilities. AB - BACKGROUND: Residential care is important for older adults, particularly for those with advanced dementia and their families. Education interventions that achieve sustainable improvement in the care of older adults are critical to quality care. There are few systematic data available regarding the educational needs of Residential Care Facility (RCF) staff and General Practitioners (GPs) relating to dementia, or the sustainability of educational interventions. We sought to determine participation in dementia education, perceived levels of current knowledge regarding dementia, perceived unmet educational needs, current barriers, facilitators and preferences for dementia education. METHODS: A mixed methods study design was utilised. A survey was distributed to a convenience sample of general practitioners, and staff in 223 consecutive residential care facilities in Perth, Western Australia. Responses were received from 102 RCF staff working in 10 facilities (out of 33 facilities who agreed to distribute the survey) and 202 GPs (19% of metropolitan GPs). Quantitative survey data were summarised descriptively and chi squared statistics were used to analyse the distribution of categorical variables. Qualitative data were collected from general practitioners, staff in residential care facilities and family carers of people with dementia utilizing individual interviews, surveys and focus groups. Qualitative data were analysed thematically. RESULTS: Among RCF staff and GPs attending RCF, participation in dementia education was high, and knowledge levels generally perceived as good. The individual experiences and needs of people with dementia and their families were emphasised. Participants identified the need for a person centred philosophy to underpin educational interventions. Limited time was a frequently mentioned barrier, especially in relation to attending dementia care education. Perceived educational needs relating to behaviours of concern, communication, knowledge regarding dementia, aspects of person centred care, system factors and the multidisciplinary team were consistently and frequently cited. Small group education which is flexible, individualized, practical and case based was sought. CONCLUSION: The effectiveness and sustainability of an educational intervention based on these findings needs to be tested. In addition, future interventions should focus on supporting cultural change to facilitate sustainable improvements in care. PMID- 19674463 TI - A gene expression profile for detection of sufficient tumour cells in breast tumour tissue: microarray diagnosis eligibility. AB - BACKGROUND: Microarray diagnostics of tumour samples is based on measurement of prognostic and/or predictive gene expression profiles. Typically, diagnostic profiles have been developed using bulk tumour samples with a sufficient amount of tumour cells (usually >50%). Consequentially, a diagnostic results depends on the minimal percentage of tumour cells within a sample. Currently, tumour cell percentage is assessed by conventional histopathological review. However, even for experienced pathologists, such scoring remains subjective and time consuming and can lead to ambiguous results. METHODS: In this study we investigated whether we could use transcriptional activity of a specific set of genes instead of histopathological review to identify samples with sufficient tumour cell content. Genome-wide gene expression measurements were used to develop a transcriptional gene profile that could accurately assess a sample's tumour cell percentage. RESULTS: Supervised analysis across 165 breast tumour samples resulted in the identification of a set of 13 genes which expression correlated with presence of tumour cells. The developed gene profile showed a high performance (AUC 0.92) for identification of samples that are suitable for microarray diagnostics. Validation on 238 additional breast tumour samples indicated a robust performance for correct classification with an overall accuracy of 91 percent and a kappa score of 0.63 (95%CI 0.47-0.73). CONCLUSION: The developed 13-gene profile provides an objective tool for assessment whether a breast cancer sample contains sufficient tumour cells for microarray diagnostics. It will improve the efficiency and throughput for diagnostic gene expression profiling as it no longer requires histopathological analysis for initial tumour percentage scoring. Such profile will also be very use useful for assessment of tumour cell percentage in biopsies where conventional histopathology is difficult, such as fine needle aspirates. PMID- 19674464 TI - Whole shaft visibility and mechanical performance for active MR catheters using copper-nitinol braided polymer tubes. AB - BACKGROUND: Catheter visualization and tracking remains a challenge in interventional MR.Active guidewires can be made conspicuous in "profile" along their whole shaft exploiting metallic core wire and hypotube components that are intrinsic to their mechanical performance. Polymer-based catheters, on the other hand, offer no conductive medium to carry radio frequency waves. We developed a new "active" catheter design for interventional MR with mechanical performance resembling braided X-ray devices. Our 75 cm long hybrid catheter shaft incorporates a wire lattice in a polymer matrix, and contains three distal loop coils in a flexible and torquable 7Fr device. We explored the impact of braid material designs on radiofrequency and mechanical performance. RESULTS: The incorporation of copper wire into in a superelastic nitinol braided loopless antenna allowed good visualization of the whole shaft (70 cm) in vitro and in vivo in swine during real-time MR with 1.5 T scanner. Additional distal tip coils enhanced tip visibility. Increasing the copper:nitinol ratio in braiding configurations improved flexibility at the expense of torquability. We found a 16 wire braid of 1:1 copper:nitinol to have the optimum balance of mechanical (trackability, flexibility, torquability) and antenna (signal attenuation) properties. With this configuration, the temperature increase remained less than 2 degrees C during real-time MR within 10 cm horizontal from the isocenter. The design was conspicuous in vitro and in vivo. CONCLUSION: We have engineered a new loopless antenna configuration that imparts interventional MR catheters with satisfactory mechanical and imaging characteristics. This compact loopless antenna design can be generalized to visualize the whole shaft of any general purpose polymer catheter to perform safe interventional procedures. PMID- 19674466 TI - Bilateral myositis ossificans of the masseter muscle after chemoradiotherapy and critical illness neuropathy--report of a rare entity and review of literature. AB - Myositis ossificans in the head and neck is a rare heterotropic bone formation within a muscle. Besides fibrodysplasia ossificans progressiva, traumatic and neurogenic forms are described in the literature. We are presenting the case of a 35-year-old female patient with a very rare form of MO of both masseter muscles after 4 weeks of intensive care because of complications (critical illness neuropathy) after chemotherapy. Therefore, special attention should be paid to surgical trauma. As in the present case, radiotherapy, long-time intubation with immobilization and critical myopathy and neuropathy can cause MO with severe problems, such as trismus and reduced mouth hygiene, which can lead to reduced quality of life. PMID- 19674465 TI - Mechanism of action of lenalidomide in hematological malignancies. AB - Immunomodulatory drugs lenalidomide and pomalidomide are synthetic compounds derived by modifying the chemical structure of thalidomide to improve its potency and reduce its side effects. Lenalidomide is a 4-amino-glutamyl analogue of thalidomide that lacks the neurologic side effects of sedation and neuropathy and has emerged as a drug with activity against various hematological and solid malignancies. It is approved by FDA for clinical use in myelodysplastic syndromes with deletion of chromosome 5q and multiple myeloma. Lenalidomide has been shown to be an immunomodulator, affecting both cellular and humoral limbs of the immune system. It has also been shown to have anti-angiogenic properties. Newer studies demonstrate its effects on signal transduction that can partly explain its selective efficacy in subsets of MDS. Even though the exact molecular targets of lenalidomide are not well known, its activity across a spectrum of neoplastic conditions highlights the possibility of multiple target sites of action. PMID- 19674468 TI - Hepatoid adenocarcinoma of the stomach - a different histology for not so different gastric adenocarcinoma: a case report. AB - Hepatoid adenocarcinoma is an extrahepatic tumor characterized by morphological similarities to hepatocellular carcinoma. Hepatoid adenocarcinoma of the stomach is a cancer with an extremely poor prognosis with few cases reported. Here, we describe a 75-year-old Spanish man referred to our hospital with a history of abdominal pain, general fatigue, anorexia and sickness. Initial study revealed anemia, and computed tomography scan and abdominal ultrasonography showed multiple metastases to the liver with hepatocellular carcinoma characteristics in a liver with no cirrhotic change. Further study included a serum level of alpha fetoprotein (AFP), which resulted markedly elevated, and a conclusive esophagogastroduodenoscopy describing an elevated tumour growing through the cardia and gastroesophageal junction with foci of necrosis and haemorrhage. Gastric biopsies of the tumor revealed poorly differenciated adenocarcinoma, with hepatoid differentiation. After a diagnosis of AFP-producing hepatoid adenocarcinoma of the stomach with multiple liver metastases was made, pallitive total gastrectomy, without liver resection, was performed. Patient recovered well after surgery, and entered into a palliative systemich chemotherapy protocol. Although this illness is recognized as having poor prognosis, the patient remains alive 8 months after the operation. Accurate diagnosis of hepatoid adenocarcinoma of the stomach is important, and should be suspected under certain circumstances. We describe this rare case of hepatoid adenocarcinoma of the stomach, and review the literature concerning the clinicopathological aspects. PMID- 19674467 TI - Psychosocial correlates of eating behavior in children and adolescents: a review. AB - BACKGROUND: Understanding the correlates of dietary intake is necessary in order to effectively promote healthy dietary behavior among children and adolescents. A literature review was conducted on the correlates of the following categories of dietary intake in children and adolescents: Fruit, Juice and Vegetable Consumption, Fat in Diet, Total Energy Intake, Sugar Snacking, Sweetened Beverage Consumption, Dietary Fiber, Other Healthy Dietary Consumption, and Other Less Healthy Dietary Consumption in children and adolescents. METHODS: Cross-sectional and prospective studies were identified from PubMed, PsycINFO and PsycArticles by using a combination of search terms. Quantitative research examining determinants of dietary intake among children and adolescents aged 3-18 years were included. The selection and review process yielded information on country, study design, population, instrument used for measuring intake, and quality of research study. RESULTS: Seventy-seven articles were included. Many potential correlates have been studied among children and adolescents. However, for many hypothesized correlates substantial evidence is lacking due to a dearth of research. The correlates best supported by the literature are: perceived modeling, dietary intentions, norms, liking and preferences. Perceived modeling and dietary intentions have the most consistent and positive associations with eating behavior. Norms, liking, and preferences were also consistently and positively related to eating behavior in children and adolescents. Availability, knowledge, outcome expectations, self-efficacy and social support did not show consistent relationships across dietary outcomes. CONCLUSION: This review examined the correlates of various dietary intake; Fruit, Juice and Vegetable Consumption, Fat in Diet, Total Energy Intake, Sugar Snacking, Sweetened Beverage Consumption, Dietary Fiber, Other Healthy Dietary Consumption, and Other Less Healthy Dietary Consumption in cross-sectional and prospective studies for children and adolescents. The correlates most consistently supported by evidence were perceived modeling, dietary intentions, norms, liking and preferences. More prospective studies on the psychosocial determinants of eating behavior using broader theoretical perspectives should be examined in future research. PMID- 19674469 TI - Intrinsic expression of host genes and intronic miRNAs in prostate carcinoma cells. AB - BACKGROUND: Recent data show aberrant and altered expression of regulatory noncoding micro (mi) RNAs in prostate cancer (PCa). A large number of miRNAs are encoded in organized intronic clusters within many protein coding genes. While expression profiling studies of miRNAs are common place, little is known about the host gene and their resident miRNAs coordinated expression in PCa cells. Furthermore, whether expression of a subset of miRNAs is distinct in androgen responsive and androgen-independent cells is not clear. Here we have examined the expression of mature miRNAs of miR 17-92, miR 106b-25 and miR 23b-24 clusters along with their host genes C13orf25, MCM7 and AMPO respectively in PCa cell lines. RESULTS: The expression profiling of miRNAs and host genes was performed in androgen-sensitive MDA PCa 2b and LNCaP as well as in androgen-refractory PC-3 and DU 145 cell culture models of PCa. No significant correlation between the miRNA expression and the intrinsic hormone-responsive property of PCa cells was observed. Androgen-sensitive MDA PCa 2b cells exhibited the highest level of expression of most miRNAs studied in this report. We found significant expression variations between host genes and their resident miRNAs. The expressions of C13orf25 and miR 17-92 cluster as well as MCM7 and miR 106b-25 cluster did not reveal statistically significant correlation, thus suggesting that host genes and resident miRNAs may be expressed independent of each other. CONCLUSION: Our results suggest that miRNA expression profiles may not predict intrinsic hormone sensitive environment of PCa cells. More importantly, our data indicate the possibility of additional novel mechanisms for intronic miRNA processing in PCa cells. PMID- 19674470 TI - New classification of maxillary ameloblastic carcinoma based on an evidence-based literature review over the last 60 years. AB - BACKGROUND: The ameloblastic carcinoma is a rare malignant odontogenic tumor which rather occurs in the mandible than in the maxilla. Its rarity and in this context somewhat speculative histopathogenesis may account for diagnostic difficulties. Current classifications do not consider benign histopathological features at the primary and malignant features at the metastatic tumour site. Based on an evidence-based literature review, a recommendation for a novel classification is presented. METHODS: An evidence-based literature review over the last 60 years regarding ameloblastic carcinoma of the maxilla was conducted. RESULTS: An overall of 26 cases were found (mean age: 54.4 (583 years); male to female ratio: 2.7 to 1). In 54% the primary diagnosis was ameloblastic carcinoma, 34.6% revealed pulmonary metastases, however, only in one patient cervical lymph node metastasis could be found. Whereas two cases did not reveal malignant histopathology at the primary, they revealed malignant features at their metastatic sites. Nineteen of 26 patients (73,1%) were controlled during a median follow-up time of 54,3 months (6 to 156 months); 6 patients died of disease after a median time of 62,7 months (7 to 156 months) after initial diagnosis. CONCLUSION: It is of utmost importance to be aware of that ameloblastomas may be capable to degenerate into a "malignant" disease with recurrence and metastasis. In addition to local long-term control, special attention should be paid to potential pulmonary involvement. PMID- 19674472 TI - Review of cigarette smoking and tuberculosis in China: intervention is needed for smoking cessation among tuberculosis patients. AB - BACKGROUND: As a risk factor of tuberculosis (TB), tobacco smoking has increased substantially over the past three decades, especially in developing countries. However, the association between smoking and TB, which has been shown to exist in different studies with different ethnic background, has not yet received sufficient attention in terms of TB care standards and research in China. METHODS: An observational study was conducted in two rural areas of China. A total of 613 TB patients frequency matched with 1226 controls were interviewed by using a structured questionnaire. The associations between cigarette smoking and risk of TB were estimated by computing odds ratios (ORs) and 95% confidence intervals (95% CIs) from logistic regression model. Patients' smoking behavior and patterns of smoking cessation were followed after TB diagnosis. Multivariate Cox proportional hazards model was applied to calculate hazard ratios (HRs) and 95% confidence intervals (95% CIs) in analyzing the risk factors for smoking relapse. The Kaplan-Meier estimate was computed to plot the ability of smoking free after cessation among different groups, with the Log-rank test being used to compare the difference. RESULTS: The proportion of cigarette smoking was 54.6% in TB cases, which was significantly higher than that in controls (45.1%) with adjusted OR of 1.93(95% CI: 1.51-2.48). Though 54.9% smokers stopped smoking after being diagnosed with TB, more than 18% relapsed during the follow-up period. The proportion of relapse was higher within 6-9 months (6%) and 12-15 months (11%) after cessation. In the Cox regression estimates adjusted for age and gender, compared with those highly educated and previously treated patients, the hazard ratios of smoking relapse were 3.48(95% CI: 1.28-9.47) for less educated (<6 years) and 4.30(95% CI: 1.01-18.30) for newly treated patients, respectively. CONCLUSION: Cigarette smoking is associated with TB in the Chinese. Interventions of smoking cessation are recommended to be included in the current TB control practice. PMID- 19674471 TI - Can cognitive enhancers reduce the risk of falls in older people with mild cognitive impairment? A protocol for a randomised controlled double blind trial. AB - BACKGROUND: Older adults with cognitive problems have a higher risk of falls, at least twice that of cognitively normal older adults. The consequences of falls in this population are very serious: fallers with cognitive problems suffer more injuries due to falls and are approximately five times more likely to be admitted to institutional care. Although the mechanisms of increased fall risk in cognitively impaired people are not completely understood, it is known that impaired cognitive abilities can reduce attentional resource allocation while walking. Since cognitive enhancers, such as cholinesterase inhibitors, improve attention and executive function, we hypothesise that cognitive enhancers may reduce fall risk in elderly people in the early stages of cognitive decline by improving their gait and balance performance due to an enhancement in attention and executive function. METHOD/DESIGN: Double blinded randomized controlled trial with 6 months follow-up in 140 older individuals with Mild Cognitive Impairment (MCI). Participants will be randomized to the intervention group, receiving donepezil, and to the control group, receiving placebo. A block randomization by four and stratification based on fall history will be performed. Primary outcomes are improvements in gait velocity and reduction in gait variability. Secondary outcomes are changes in the balance confidence, balance sway, attention, executive function, and number of falls. DISCUSSION: By characterizing and understanding the effects of cognitive enhancers on fall risk in older adults with cognitive impairments, we will be able to pave the way for a new approach to fall prevention in this population. This RCT study will provide, for the first time, information regarding the effect of a medication designed to augment cognitive functioning have on the risk of falls in older adults with Mild Cognitive Impairment. We expect a significant reduction in the risk of falls in this vulnerable population as a function of the reduced gait variability achieved by treatment with cognitive enhancers. This study may contribute to a new approach to prevent and treat fall risk in seniors in early stages of dementia. TRIAL REGISTRATION: The protocol for this study is registered with the Clinical Trials Registry, identifier number: NCT00934531 http://www.clinicaltrials.gov. PMID- 19674473 TI - Methamphetamine use and rates of incarceration among street-involved youth in a Canadian setting: a cross-sectional analysis. AB - BACKGROUND: Given concerns over rising use of methamphetamine, especially among street-involved youth, and the links between exposure to the correctional system and the production of drug-related harm, we sought to assess the relationship between ever using methamphetamine and reporting ever being incarcerated in the At-Risk Youth Survey (ARYS) in Vancouver, Canada. METHODS: The relationship between ever being imprisoned and ever using methamphetamine was estimated using a multivariate logistic regression analysis while also considering potentially confounding secondary demographic, social and behavioural variables. RESULTS: Of the 478 youth recruited into ARYS between September 2005 and October 2006, 385 (80.5%) reported ever being incarcerated overnight or longer. In the multivariate model, methamphetamine use was independently associated with ever being incarcerated (Adjusted Odds Ratio: 1.79, 95% Confidence Interval [CI]: 1.03 - 3.13). CONCLUSION: Incarceration was very common in this cohort and strongly linked with ever using methamphetamine. This finding is of concern and, along with the previously identified risks of drug-related harm associated with incarceration, supports the development of novel public policy, such as community based drug treatment, to address the use of methamphetamine among street youth. PMID- 19674474 TI - Traditional Chinese Medicines in the treatment of hepatocellular cancers: a systematic review and meta-analysis. AB - BACKGROUND: Liver cancer is a common malignancy with a high mortality rate. Given the poor prognosis associated with this cancer, many patients seek additional therapies that may improve quality of life or survival. Several Traditional Chinese Medicines (TCM) have been evaluated in clinical trials, but little is known about them outside of China. METHODS: We searched independently and in duplicate 8 electronic databases, including 2 Chinese language databases, until February 2009. We included any randomized clinical trials (RCT) evaluating a TCM oral preparation for the treatment of hepatocellular cancers. We abstracted data on survival, tumor response, and performance scores. We conducted a random effects meta-analysis and applied a meta-regression analysis. RESULTS: We included 45 RCTs (n = 3,236). All studies employed an active control group. In general, the reporting of methodological issues was poor. We analyzed data from 37 trials reporting on complete response effects score (Relative Risk [RR] of 1.26 (95 CI, 1.04-1.52, P = 0.01, I2 = 0%, P = 0.99). Products containing ginseng, astragalus and mylabris had a larger treatment effect (OR 1.34, 95% CI, 1.04-1.71, P = 0.01) than the pooled broad estimate, also the case for astragalus based treatments (OR 1.35, 95% CI, 1.001-1.80. P = 0.048). We examined survival rates and pooled 15 studies reporting on 6 month outcomes (RR 1.10, 95% CI, 1.04 1.15, P = < 0.0001, I2 = 0%, P = 0.60). This effect was consistent at other prospective dates, including 12 months (22 trials, RR 1.26, 95% CI, 1.17-1.36, P = < 0.0001, I2 = 7%, P = 0.36), 24 months (15 trials, 1.72, 95% CI, 1.40-2.03, P = < 0.0001, I2 = 0%, P = 0.75); and, at 36 months (8 trials, RR 2.40, 95% CI, 1.65-3.49, P = < 0.0001, I2 = 0%, P = 0.62). LIMITATIONS: All included trials were conducted in China where emerging evidence suggests many RCTs are not, in fact, randomized. Publication bias may exist, favouring positive reports. CONCLUSION: Our meta-analysis displays compelling evidence of effectiveness for hepatocellular cancers that should be evaluated in high-quality and transparent clinical trials. PMID- 19674475 TI - A mutation in CTSK gene in an autosomal recessive pycnodysostosis family of Pakistani origin. AB - BACKGROUND: Pycnodysostosis is a rare autosomal recessive skeletal dysplasia characterized by short stature, osteosclerosis, acro-osteolysis, frequent fractures and skull deformities. Mutations in the gene encoding cathepsin K (CTSK), a lysosomal cysteine protease, have been found to be responsible for this disease. OBJECTIVES: To identify pathogenic mutation in a consanguineous Pakistani family with 3 affected individuals demonstrating autosomal recessive pycnodysostosis. METHODS: Genotyping of 10 members of the family, including three affected and seven unaffected individuals was carried out by using polymorphic markers D1S442, D1S498, and D1S305, which are closely linked to the CTSK gene on chromosome 1q21. To screen for mutations in the CTSK gene, all of its exons and splice junctions were PCR amplified from genomic DNA and sequenced directly in an ABI Prism 310 automated sequencer. RESULTS: Genotyping results showed linkage of the pycnodysostosis Pakistani family to the CTSK locus. Sequence analysis of the CTSK gene revealed homozygosity for a missense mutation (A277V) in the affected individuals. CONCLUSION: We describe a missense mutation in the CTSK gene in a Pakistani family affected with autosomal recessive pycnodysostosis. Our study strengthens the role of this particular mutation in the pathogenesis of pycnodysostosis and suggests its prevalence in Pakistani patients. PMID- 19674477 TI - Comparative evaluation of the Ifakara tent trap-B, the standardized resting boxes and the human landing catch for sampling malaria vectors and other mosquitoes in urban Dar es Salaam, Tanzania. AB - BACKGROUND: Frequent, sensitive and accurate sampling of Anopheles mosquitoes is a prerequisite for effective management of malaria vector control programmes. The most reliable existing means to measure mosquito density is the human landing catch (HLC). However, the HLC technique raises major ethical concerns because of the necessity to expose humans to vectors of malaria and a variety of other pathogens. Furthermore, it is a very arduous undertaking that requires intense supervision, which is severely limiting in terms of affordability and sustainability. METHODS: A community-based, mosquito sampling protocol, using the Ifakara tent trap-B (ITT-B) and standardized resting boxes (SRB), was developed and evaluated in terms of the number and sample composition of mosquitoes caught by each, compared to rigorously controlled HLC. Mosquitoes were collected once and three times every week by the HLC and the alternative methods, respectively, in the same time and location. RESULTS: Overall, the three traps caught 44,848 mosquitoes. The ITT-B, HLC and SRB caught 168, 143 and 46 Anopheles gambiae s.l. as well as 26,315, 13,258 and 4,791 Culex species respectively. The ITT-B was three- and five-times cheaper than the HLC per mosquito caught for An. gambiae and Cx. Species, respectively. Significant correlations between the numbers caught by HLC and ITT-B were observed for both An. gambiae s.l. (P < 0.001) and Cx. species (P = 0.003). Correlation between the catches with HLC and SRB were observed for Cx. species (P < 0.001) but not An. gambiae s.l. (P = 0.195), presumably because of the low density of the latter. Neither ITT-B nor SRB exhibited any obvious density dependence for sampling the two species. CONCLUSION: SRBs exhibited poor sensitivity for both mosquito taxa and are not recommended in this setting. However, this protocol is affordable and effective for routine use of the ITT-B under programmatic conditions. Nevertheless, it is recommended that the trap and the protocol be evaluated further at full programmatic scales to establish effectiveness under fully representative conditions of routine practice. PMID- 19674476 TI - Validation of a short form Wisconsin Upper Respiratory Symptom Survey (WURSS-21). AB - BACKGROUND: The Wisconsin Upper Respiratory Symptom Survey (WURSS) is an illness specific health-related quality-of-life questionnaire outcomes instrument. OBJECTIVES: Research questions were: 1) How well does the WURSS-21 assess the symptoms and functional impairments associated with common cold? 2) How well can this instrument measure change over time (responsiveness)? 3) What is the minimal important difference (MID) that can be detected by the WURSS-21? 4) What are the descriptive statistics for area under the time severity curve (AUC)? 5) What sample sizes would trials require to detect MID or AUC criteria? 6) What does factor analysis tell us about the underlying dimensional structure of the common cold? 7) How reliable are items, domains, and summary scores represented in WURSS? 8) For each of these considerations, how well does the WURSS-21 compare to the WURSS-44, Jackson, and SF-8? STUDY DESIGN AND SETTING: People with Jackson defined colds were recruited from the community in and around Madison, Wisconsin. Participants were enrolled within 48 hours of first cold symptom and monitored for up to 14 days of illness. Half the sample filled out the WURSS-21 in the morning and the WURSS-44 in the evening, with the other half reversing the daily order. External comparators were the SF-8, a 24-hour recall general health measure yielding separate physical and mental health scores, and the eight-item Jackson cold index, which assesses symptoms, but not functional impairment or quality of life. RESULTS: In all, 230 participants were monitored for 2,457 person-days. Participants were aged 14 to 83 years (mean 34.1, SD 13.6), majority female (66.5%), mostly white (86.0%), and represented substantive education and income diversity. WURSS-21 items demonstrated similar performance when embedded within the WURSS-44 or in the stand-alone WURSS-21. Minimal important difference (MID) and Guyatt's responsiveness index were 10.3, 0.71 for the WURSS-21 and 18.5, 0.75 for the WURSS-44. Factorial analysis suggested an eight dimension structure for the WURSS-44 and a three dimension structure for the WURSS-21, with composite reliability coefficients ranging from 0.87 to 0.97, and Cronbach's alpha ranging from 0.76 to 0.96. Both WURSS versions correlated significantly with the Jackson scale (W-21 R=0.85; W-44 R=0.88), with the SF-8 physical health (W-21 R=-0.79; W-44 R=-0.80) and SF-8 mental health (W-21 R=-0.55; W-44 R=-0.60). CONCLUSION: The WURSS-44 and WURSS-21 perform well as illness-specific quality-of life evaluative outcome instruments. Construct validity is supported by the data presented here. While the WURSS-44 covers more symptoms, the WURSS-21 exhibits similar performance in terms of reliability, responsiveness, importance-to patients, and convergence with other measures. PMID- 19674478 TI - EvoRSR: an integrated system for exploring evolution of RNA structural robustness. AB - BACKGROUND: Robustness, maintaining a constant phenotype despite perturbations, is a fundamental property of biological systems that is incorporated at various levels of biological complexity. Although robustness has been frequently observed in nature, its evolutionary origin remains unknown. Current hypotheses suggest that robustness originated as a direct consequence of natural selection, as an intrinsic property of adaptations, or as a congruent correlate of environment robustness. To elucidate the evolutionary origins of robustness, a convenient computational package is strongly needed. RESULTS: In this study, we developed the open-source integrated system EvoRSR (Evolution of RNA Structural Robustness) to explore the evolution of robustness based on biologically important landscapes induced by RNA folding. EvoRSR is object-oriented, modular, and freely available at http://biotech.bmi.ac.cn/EvoRSR under the GNU/GPL license. We present an overview of EvoRSR package and illustrate its features with the miRNA gene cel mir-357. CONCLUSION: EvoRSR is a novel and flexible package for exploring the evolution of robustness. Accordingly, EvoRSR can be used for future studies to investigate the evolution and origin of robustness and to address other common questions about robustness. While the current EvoRSR environment is a versatile analysis framework, future versions can include features to enhance evolutionary studies of robustness. PMID- 19674480 TI - ArachnoServer: a database of protein toxins from spiders. AB - BACKGROUND: Venomous animals incapacitate their prey using complex venoms that can contain hundreds of unique protein toxins. The realisation that many of these toxins may have pharmaceutical and insecticidal potential due to their remarkable potency and selectivity against target receptors has led to an explosion in the number of new toxins being discovered and characterised. From an evolutionary perspective, spiders are the most successful venomous animals and they maintain by far the largest pool of toxic peptides. However, at present, there are no databases dedicated to spider toxins and hence it is difficult to realise their full potential as drugs, insecticides, and pharmacological probes. DESCRIPTION: We have developed ArachnoServer, a manually curated database that provides detailed information about proteinaceous toxins from spiders. Key features of ArachnoServer include a new molecular target ontology designed especially for venom toxins, the most up-to-date taxonomic information available, and a powerful advanced search interface. Toxin information can be browsed through dynamic trees, and each toxin has a dedicated page summarising all available information about its sequence, structure, and biological activity. ArachnoServer currently manages 567 protein sequences, 334 nucleic acid sequences, and 51 protein structures. CONCLUSION: ArachnoServer provides a single source of high-quality information about proteinaceous spider toxins that will be an invaluable resource for pharmacologists, neuroscientists, toxinologists, medicinal chemists, ion channel scientists, clinicians, and structural biologists. ArachnoServer is available online at http://www.arachnoserver.org. PMID- 19674479 TI - Multiple functions of precursor BDNF to CNS neurons: negative regulation of neurite growth, spine formation and cell survival. AB - BACKGROUND: Proneurotrophins and mature neurotrophins elicit opposite effects via the p75 neurotrophin receptor (p75(NTR)) and Trk tyrosine kinase receptors, respectively; however the molecular roles of proneurotrophins in the CNS are not fully understood. RESULTS: Based on two rare single nucleotide polymorphisms (SNPs) of the human brain-derived neurotrophic factor (BDNF) gene, we generated R125M-, R127L- and R125M/R127L-BDNF, which have amino acid substitution(s) near the cleavage site between the pro- and mature-domain of BDNF. Western blot analyses demonstrated that these BDNF variants are poorly cleaved and result in the predominant secretion of proBDNF. Using these cleavage-resistant proBDNF (CR proBDNF) variants, the molecular and cellular roles of proBDNF on the CNS neurons were examined. First, CR-proBDNF showed normal intracellular distribution and secretion in cultured hippocampal neurons, suggesting that inhibition of proBDNF cleavage does not affect intracellular transportation and secretion of BDNF. Second, we purified recombinant CR-proBDNF and tested its biological effects using cultured CNS neurons. Treatment with CR-proBDNF elicited apoptosis of cultured cerebellar granule neurons (CGNs), while treatment with mature BDNF (matBDNF) promoted cell survival. Third, we examined the effects of CR-proBDNF on neuronal morphology using more than 2-week cultures of basal forebrain cholinergic neurons (BFCNs) and hippocampal neurons. Interestingly, in marked contrast to the action of matBDNF, which increased the number of cholinergic fibers and hippocampal dendritic spines, CR-proBDNF dramatically reduced the number of cholinergic fibers and hippocampal dendritic spines, without affecting the survival of these neurons. CONCLUSION: These results suggest that proBDNF has distinct functions in different populations of CNS neurons and might be responsible for specific physiological cellular processes in the brain. PMID- 19674482 TI - Complex correlation measure: a novel descriptor for Poincare plot. AB - BACKGROUND: Poincare plot is one of the important techniques used for visually representing the heart rate variability. It is valuable due to its ability to display nonlinear aspects of the data sequence. However, the problem lies in capturing temporal information of the plot quantitatively. The standard descriptors used in quantifying the Poincare plot (SD1, SD2) measure the gross variability of the time series data. Determination of advanced methods for capturing temporal properties pose a significant challenge. In this paper, we propose a novel descriptor "Complex Correlation Measure (CCM)" to quantify the temporal aspect of the Poincare plot. In contrast to SD1 and SD2, the CCM incorporates point-to-point variation of the signal. METHODS: First, we have derived expressions for CCM. Then the sensitivity of descriptors has been shown by measuring all descriptors before and after surrogation of the signal. For each case study, lag-1 Poincare plots were constructed for three groups of subjects (Arrhythmia, Congestive Heart Failure (CHF) and those with Normal Sinus Rhythm (NSR)), and the new measure CCM was computed along with SD1 and SD2. ANOVA analysis distribution was used to define the level of significance of mean and variance of SD1, SD2 and CCM for different groups of subjects. RESULTS: CCM is defined based on the autocorrelation at different lags of the time series, hence giving an in depth measurement of the correlation structure of the Poincare plot. A surrogate analysis was performed, and the sensitivity of the proposed descriptor was found to be higher as compared to the standard descriptors. Two case studies were conducted for recognizing arrhythmia and congestive heart failure (CHF) subjects from those with NSR, using the Physionet database and demonstrated the usefulness of the proposed descriptors in biomedical applications. CCM was found to be a more significant (p = 6.28E-18) parameter than SD1 and SD2 in discriminating arrhythmia from NSR subjects. In case of assessing CHF subjects also against NSR, CCM was again found to be the most significant (p = 9.07E-14). CONCLUSION: Hence, CCM can be used as an additional Poincare plot descriptor to detect pathology. PMID- 19674483 TI - Effect of two yoga-based relaxation techniques on memory scores and state anxiety. AB - BACKGROUND: A yoga practice involving cycles of yoga postures and supine rest (called cyclic meditation) was previously shown to improve performance in attention tasks more than relaxation in the corpse posture (shavasana). This was ascribed to reduced anxiety, though this was not assessed. METHODS: In fifty seven male volunteers (group average age +/- S.D., 26.6 +/- 4.5 years) the immediate effect of two yoga relaxation techniques was studied on memory and state anxiety. All participants were assessed before and after (i) Cyclic meditation (CM) practiced for 22:30 minutes on one day and (ii) an equal duration of Supine rest (SR) or the corpse posture (shavasana), on another day. Sections of the Wechsler memory scale (WMS) were used to assess; (i) attention and concentration (digit span forward and backward), and (ii) associate learning. State anxiety was assessed using Spielberger's State-Trait Anxiety Inventory (STAI). RESULTS: There was a significant improvement in the scores of all sections of the WMS studied after both CM and SR, but, the magnitude of change was more after CM compared to after SR. The state anxiety scores decreased after both CM and SR, with a greater magnitude of decrease after CM. There was no correlation between percentage change in memory scores and state anxiety for either session. CONCLUSION: A cyclical combination of yoga postures and supine rest in CM improved memory scores immediately after the practice and decreased state anxiety more than rest in a classical yoga relaxation posture (shavasana). PMID- 19674481 TI - A dual propagation contours technique for semi-automated assessment of systolic and diastolic cardiac function by CMR. AB - BACKGROUND: Although cardiovascular magnetic resonance (CMR) is frequently performed to measure accurate LV volumes and ejection fractions, LV volume-time curves (VTC) derived ejection and filling rates are not routinely calculated due to lack of robust LV segmentation techniques. VTC derived peak filling rates can be used to accurately assess LV diastolic function, an important clinical parameter. We developed a novel geometry-independent dual-contour propagation technique, making use of LV endocardial contours manually drawn at end systole and end diastole, to compute VTC and measured LV ejection and filling rates in hypertensive patients and normal volunteers. METHODS: 39 normal volunteers and 49 hypertensive patients underwent CMR. LV contours were manually drawn on all time frames in 18 normal volunteers. The dual-contour propagation algorithm was used to propagate contours throughout the cardiac cycle. The results were compared to those obtained with single-contour propagation (using either end-diastolic or end systolic contours) and commercially available software. We then used the dual contour propagation technique to measure peak ejection rate (PER) and peak early diastolic and late diastolic filling rates (ePFR and aPFR) in all normal volunteers and hypertensive patients. RESULTS: Compared to single-contour propagation methods and the commercial method, VTC by dual-contour propagation showed significantly better agreement with manually-derived VTC. Ejection and filling rates by dual-contour propagation agreed with manual (dual-contour - manual PER: -0.12 +/- 0.08; ePFR: -0.07 +/- 0.07; aPFR: 0.06 +/- 0.03 EDV/s, all P = NS). However, the time for the manual method was approximately 4 hours per study versus approximately 7 minutes for dual-contour propagation. LV systolic function measured by LVEF and PER did not differ between normal volunteers and hypertensive patients. However, ePFR was lower in hypertensive patients vs. normal volunteers, while aPFR was higher, indicative of altered diastolic filling rates in hypertensive patients. CONCLUSION: Dual-propagated contours can accurately measure both systolic and diastolic volumetric indices that can be applied in a routine clinical CMR environment. With dual-contour propagation, the user interaction that is routinely performed to measure LVEF is leveraged to obtain additional clinically relevant parameters. PMID- 19674484 TI - Exploration of the beliefs and experiences of Aboriginal people with cancer in Western Australia: a methodology to acknowledge cultural difference and build understanding. AB - BACKGROUND: Aboriginal Australians experience poorer outcomes, and are 2.5 times more likely to die from cancer than non-Aboriginal people, even after adjustment for stage of diagnosis, cancer treatment and comorbidities. They are also less likely to present early as a result of symptoms and to access treatment. Psycho social factors affect Aboriginal people's willingness and ability to participate in cancer-related screening and treatment services, but little exploration of this has occurred within Australia to date. The current research adopted a phenomenological qualitative approach to understand and explore the lived experiences of Aboriginal Australians with cancer and their beliefs and understanding around this disease in Western Australia (WA). This paper details considerations in the design and process of conducting the research. METHODS/DESIGN: The National Health and Medical Research Council (NHMRC) guidelines for ethical conduct of Aboriginal research were followed. Researchers acknowledged the past negative experiences of Aboriginal people with research and were keen to build trust and relationships prior to conducting research with them. Thirty in-depth interviews with Aboriginal people affected by cancer and twenty with health service providers were carried out in urban, rural and remote areas of WA. Interviews were audio-recorded, transcribed verbatim and coded independently by two researchers. NVivo7 software was used to assist data management and analysis. Participants' narratives were divided into broad categories to allow identification of key themes and discussed by the research team. DISCUSSION AND CONCLUSION: Key issues specific to Aboriginal research include the need for the research process to be relationship-based, respectful, culturally appropriate and inclusive of Aboriginal people. Researchers are accountable to both participants and the wider community for reporting their findings and for research translation so that the research outcomes benefit the Aboriginal community. There are a number of factors that influence whether the desired level of engagement can be achieved in practice. These include the level of resourcing for the project and the researchers' efforts to ensure dissemination and research translation; and the capacity of the Aboriginal community to engage with research given other demands upon their time. PMID- 19674485 TI - Cost-effectiveness of a structured progressive task-oriented circuit class training programme to enhance walking competency after stroke: the protocol of the FIT-Stroke trial. AB - BACKGROUND: Most patients who suffer a stroke experience reduced walking competency and health-related quality of life (HRQoL). A key factor in effective stroke rehabilitation is intensive, task-specific training. Recent studies suggest that intensive, patient-tailored training can be organized as a circuit with a series of task-oriented workstations. Primary aim of the FIT-Stroke trial is to evaluate the effects and cost-effectiveness of a structured, progressive task-oriented circuit class training (CCT) programme, compared to usual physiotherapeutic care during outpatient rehabilitation in a rehabilitation centre. The task-oriented CCT will be applied in groups of 4 to 6 patients. Outcome will be defined in terms of gait and gait-related ADLs after stroke. The trial will also investigate the generalizability of treatment effects of task oriented CCT in terms of perceived fatigue, anxiety, depression and perceived HRQoL. METHODS/DESIGN: The multicentre single-blinded randomized trial will include 220 stroke patients discharged to the community from inpatient rehabilitation, who are able to communicate and walk at least 10 m without physical, hands-on assistance. After discharge from inpatient rehabilitation, patients in the experimental group will receive task-oriented CCT two times a week for 12 weeks at the physiotherapy department of the rehabilitation centre. Control group patients will receive usual individual, face-to-face, physiotherapy. Costs will be evaluated by having each patient keep a cost diary for the first 24 weeks after randomisation. Primary outcomes are the mobility part of the Stroke Impact Scale (SIS-3.0) and the EuroQol. Secondary outcomes are the other domains of SIS-3.0, lower limb muscle strength, walking endurance, gait speed, balance, confidence not to fall, instrumental ADL, fatigue, anxiety, depression and HRQoL. DISCUSSION: Based on assumptions about the effect of intensity of practice and specificity of treatment effects, FIT-Stroke will address two key aims. The first aim is to investigate the effects of task oriented CCT on walking competency and HRQoL compared to usual face-to-face physiotherapy. The second aim is to reveal the cost-effectiveness of task oriented CCT in the first 6 months post stroke. Both aims were recently recommended as priorities by the American Hearth Association and Stroke Council. PMID- 19674486 TI - Vitamin B12 status in patients of Turkish and Dutch descent with depression: a comparative cross-sectional study. AB - BACKGROUND: Studies have shown a clear relationship between depressive disorders and vitamin B12 deficiency. Gastroenteritis and Helicobacter pylori infections can cause vitamin B12 deficiency. Helicobacter pylori infections are not uncommon among people of Turkish descent in The Netherlands. AIM: To examine the frequency of vitamin B12 deficiency in depressive patients of Turkish descent and compare it to the frequency of vitamin B12 deficiency in depressive patients of Dutch descent. METHODS: The present study is a comparative cross-sectional study of 47 patients of Turkish descent and 28 of Dutch descent. The depressive disorder diagnosis and differential diagnosis were made using the Structured Clinical Interview for the Diagnostic and Statistical Manual of Mental Disorders, fourth edition text revision (SCID). The severity of the depressive symptoms was determined using the Beck Depression Inventory (BDI) and the 21-item Hamilton Depression Rating Scale (HAM-D-21). Serum baseline vitamin B6 and B12, folic acid and total serum homocysteine (tHcy) levels were measured. RESULTS: The average ages of the patients of Turkish and Dutch descent were 40.57 and 44.75 years, respectively. There were no demonstrable differences between the serum vitamin B6, folic acid and tHcy levels in the two groups. The serum vitamin B12 levels were however clearly lower in the patients of Turkish descent than in those of Dutch descent. Vitamin B12 deficiency was however observed in 14 patients of Turkish descent and 1 of Dutch descent. This difference was significant. On the BDI, the patients of Turkish descent scored significantly higher than those of Dutch descent. Patients with vitamin B12 deficiency and those with hyperhomocysteinaemia had a significantly higher BDI score than patients with normal vitamin B12 and homocysteine levels. No relationship was observed with vitamin B12 and tHcy. CONCLUSION: Vitamin B12 deficiency occurs more frequently in depressive patients of Turkish than of Dutch descent. This is why it is advisable to test the vitamin B12 serum level in depressive patients of Turkish descent. PMID- 19674487 TI - Human IgG response to a salivary peptide, gSG6-P1, as a new immuno epidemiological tool for evaluating low-level exposure to Anopheles bites. AB - BACKGROUND: Human populations exposed to low malaria transmission present particular severe risks of malaria morbidity and mortality. In addition, in a context of low-level exposure to Anopheles vector, conventional entomological methods used for sampling Anopheles populations are insufficiently sensitive and probably under-estimate the real risk of malaria transmission. The evaluation of antibody (Ab) responses to arthropod salivary proteins constitutes a novel tool for estimating exposure level to insect bites. In the case of malaria, a recent study has shown that human IgG responses to the gSG6-P1 peptide represented a specific biomarker of exposure to Anopheles gambiae bites. The objective of this study was to investigate if this biomarker can be used to estimate low-level exposure of individuals to Anopheles vector. METHODS: The IgG Ab level to gSG6-P1 was evaluated at the peak and at the end of the An. gambiae exposure season in children living in Senegalese villages, where the Anopheles density was estimated to be very low by classical entomological trapping but where malaria transmission occurred during the studied season. RESULTS: Specific IgG responses to gSG6-P1 were observed in children exposed to very low-level of Anopheles bites. In addition, a significant increase in the specific IgG Ab level was observed during the Anopheles exposure season whereas classical entomological data have reported very few or no Anopheles during the studied period. Furthermore, this biomarker may also be applicable to evaluate the heterogeneity of individual exposure. CONCLUSION: The results strengthen the hypothesis that the evaluation of IgG responses to gSG6-P1 during the season of exposure could reflect the real human contact with anthropophilic Anopheles and suggest that this biomarker of low exposure could be used at the individual level. This promising immuno epidemiological marker could represent a useful tool to assess the risk to very low exposure to malaria vectors as observed in seasonal, urban, altitude or travellers contexts. In addition, this biomarker could be used for the surveillance survey after applying anti-vector strategy. PMID- 19674489 TI - Abstracts of the 4th International Conference of cGMP Generators, Effectors and Therapeutic Implications. Regensburg, Germany. June 19-21, 2009. PMID- 19674490 TI - Correspondence of continuous interstitial glucose measurement against arterialised and capillary glucose following an oral glucose tolerance test in healthy volunteers. AB - The aim of the present study was to validate the Glucoday continuous interstitial ambulatory glucose-monitoring device (AGD) against plasma glucose measured from arterialised venous (AV) and glucose from capillary whole blood (finger prick, FP) in non-diabetic subjects in response to an oral glucose tolerance test. Fifteen healthy overweight men (age 30-49 years, BMI 26-31 kg/m2) participated. Glucose levels were measured before, during and after consumption of an oral 75 g glucose load using twelve FP samples and forty-four 1 ml AV blood samples during 180 min. Interstitial glucose was measured via the AGD. Three venous samples for fasting insulin were taken to estimate insulin resistance. Profiles of AGD, AV and FP glucose were generated for each participant. Glucose values for each minute of the measurement period were interpolated using a locally weighted scatterplot smoother. Data were compared using Bland-Altman plots that showed good correspondence between all pairs of measurements. Concordance between the three methods was 0.8771 (Kendall's W, n 15, P < 0.001). Concordance was greater between AV and FP (W = 0.9696) than AGD and AV (W = 0.8770) or AGD and FP (W = 0.8764). Analysis of time to peak glucose indicated that AGD measures lagged approximately 15 min behind FP and AV measures. Percent body fat was significantly correlated with time to peak glucose levels for each measure, while BMI and estimated insulin resistance (homeostatic model assessment, HOMA) were not. In conclusion, AGD shows good correspondence with FP and AV glucose measures in response to a glucose load with a 15 min time lag. Taking this into account, AGD has potential application in nutrition and behaviour studies. PMID- 19674491 TI - Hamburger high in total, saturated and trans-fatty acids decreases HDL cholesterol and LDL particle diameter, and increases TAG, in mildly hypercholesterolaemic men. AB - The consumption of high-fat hamburger enriched with SFA and trans-fatty acids may increase risk factors for coronary vascular disease, whereas hamburger enriched with MUFA may have the opposite effect. Ten mildly hypercholesterolaemic men consumed five, 114 g hamburger patties per week for two consecutive phases. Participants consumed high-SFA hamburger (MUFA:SFA = 0.95; produced from pasture fed cattle) for 5 weeks, consumed their habitual diets for 3 weeks and then consumed high-MUFA hamburger (MUFA:SFA = 1.31; produced from grain-fed cattle) for 5 weeks. These MUFA:SFA ratios were typical of ranges observed for retail ground beef. Relative to habitual levels and levels during the high-MUFA phase, the high-SFA hamburger: increased plasma palmitic acid, palmitoleic acid and TAG (P < 0.01); decreased HDL cholesterol (HDL-C) and LDL particle diameter percentile distributions (P < 0.05); and had no effect on LDL cholesterol or plasma glucose (P>0.10). Plasma palmitoleic acid was positively correlated with TAG (r 0.90), VLDL cholesterol (r 0.73) and the LDL:HDL ratio (r 0.45), and was negatively correlated with plasma HDL-C (r - 0.58), whereas plasma palmitic, stearic and oleic acids were negatively correlated with LDL particle diameter (all P 280mg/dl): 14.30+/-8.36% (n=253). A significant inverse relationship was found between the baseline plasma vitamin C concentrations and mean percent change in total cholesterol from baseline (r = 0.500, p<0.005). It was also observed that the high and severe baseline cholesterol groups possessed lower baseline plasma vitamin C concentrations than those in the normal cholesterol groups (0.79 and 0.55 versus 1.24 mg/dl respectively). CONCLUSION: This finding strengthens the hypothesis that the cholesterol lowering and cardio-protective benefit of vitamin C supplementation may be in its ability to elevate plasma vitamin C concentrations in those patients who initially possess lower than normal vitamin C plasma concentrations. PMID- 19674667 TI - Resolution of anovulation infertility using neuro emotional technique: a report of 3 cases. AB - BACKGROUND: The female menstrual cycle is a complicated interaction of hormonal messages that are under the control of the Hypothalamic-Pituitary-Ovarian axis. Dysfunction in this axis can lead to anovulation and infertility. Stress has the potential to produce such dysfunction. OBJECTIVES: To review the normal menstrual cycle, and present a number of case studies on how the stress- reducing technique of Neuro Emotional Technique (NET) successfully aided the fertility of a number of female patients by resolving anovulation/menstrual irregularity. METHODS: Three chronic anovulating, infertile patients underwent NET. A visual analog scale was used to evaluate the effectiveness of the intervention. OUTCOMES: Anovulating patients started to ovulate following a series of treatments. Initial visual analog scale (VAS) scale on menstrual irregularity was rated 10 out of a possible 10 (anovulation) for all patients. After treatment, these 3 patients rated 0 out of 10 on the VAS scale and had fallen pregnant with subsequent birthing. A discussion of the potential link between stress and anovulation through altered gonadotropin releasing hormone pulsitile activity and how the use of NET may have resolved the anovulation seen in these 3 patients is presented. CONCLUSION: The success attributed to the NET intervention and the resumption of ovulation warrant the need for further research involving long term prospective randomized controlled trial experiments to determine a direct causal relationship. PMID- 19674669 TI - Modification of a local smoking ordinance: a case-report of chiropractic health advocacy. AB - BACKGROUND: Health Promotion may involve assessing the community and assisting in the formation of coalitions that empower a community to reduce health risks. OBJECTIVE: To describe an intervention lead by a doctor of chiropractic to reduce exposure to environmental tobacco smoke in one rural southeast Alabama town. DISCUSSION: A coalition influenced the city council to modify an existing ordinance governing smoking in public places. As a secondary goal, the coalition hoped to use the local media to significantly increase public awareness of the dangers of ETS through positive press coverage of their efforts and general media advocacy. City councilmen and the local media were involved in the coalition to use a political process to change the ordinance. Ten months after initiating the project, the existing ordinance was modified. CONCLUSION: Doctors of chiropractic involved as health advocates were able to change the local ordinance pertaining to indoor smoking. PMID- 19674668 TI - Proposed mechanisms and treatment strategies for motion sickness disorder: a case series. AB - OBJECTIVE: To present an overview of symptomatic motion sickness disorder and to propose allopathic and chiropractic approaches for treatment. CLINICAL FEATURES: Three representative cases are presented involving patients with motion sickness. All 3 patients had suffered from this condition throughout their lives. A discussion of the hypothesis of sensory conflict as a causative factor in cases of motion sickness is offered. INTERVENTION AND OUTCOME: Following methods founded in applied kinesiology and using spinal and cranial manipulative treatment, the patients were able to travel long distances without nausea, sickness, or dizziness. The evaluation of these patients' responses to treatment was determined by the doctor's observation, the patients' subjective description of symptoms while riding in a motor vehicle, a visual analog scale for neck and associated pain, and applied kinesiology chiropractic physical assessment tools. CONCLUSION: Further studies into chiropractic manipulative treatments for sensory conflict and proprioceptive dysfunctions associated with the problem of motion sickness are indicated. The hypothesis of sensory conflict as the cause of motion sickness should be explored more fully by other chiropractic physicians and researchers. PMID- 19674671 TI - Abdominal calcifications and diagnostic imaging decision making: a topic review. AB - OBJECTIVE: To review commonly encountered calcifications found within the abdomen as seen on the lumbar spine radiograph and to determine which advanced imaging modality is best to thoroughly assess the patient. METHODS: Searches of electronic databases and textbooks were conducted to construct this narrative overview. DISCUSSION: By categorizing the type of calcification and localizing it anatomically, most often a definitive diagnosis can be reached. Two commonly encountered conditions, abdominal aortic aneurysms and urinary calculi, are used to compare the main advanced imaging modalities (diagnostic ultrasound and computed tomography) used to further assess abdominal calcifications. CONCLUSION: In most circumstances, either diagnostic ultrasound or computed tomography will establish a definitive diagnosis and offer thorough imaging assessment for abdominal calcifications. PMID- 19674670 TI - Exercise therapy for low back pain: a narrative review of the literature. AB - PURPOSE: Exercise therapy has become an expected modality in the treatment of low back pain but there are no clear evidence based guidelines available. The purpose of this review is to analyze the available literature in regard to demonstrated efficacy of various types of exercise therapy for low back pain. METHOD: A search was made on PubMed for exercise and low back pain that found 28 citations in English including 23 randomized controlled clinical trials published between 1993 and 2004 and 5 literature reviews published in 2000 through 2002. RESULTS: Previous reviews have stated that the literature does not support the use of exercise therapy for acute low back pain but it does give qualified support to exercise therapy for chronic low back pain. More recent studies have divided subjects based on functional rather than temporal characteristics, specifically, taking into consideration the directional preference of the patient and addressing muscle groups that augment stabilization of the "neutral zone." Such approaches yield effective clinical response in both acute and chronic patients within 2 weeks with 6 visits and provide lasting benefit up to 3 years after intervention. CONCLUSIONS: By identifying specific functional characteristics of individual patients, providers should be able to design exercise therapies that will provide more effective clinical intervention for low back pain patients. PMID- 19674672 TI - Taking nutritional supplements for three months reduced blood pressure but not blood lipid levels in students. AB - OBJECTIVE: To study the effects of a nutritional supplement on cardiovascular health determined by blood and saliva biochemistry and heart rate variability (HRV) in asymptomatic college students and faculty members. METHODS: Forty subjects were recruited in the study via a school wide email notification and through personal contacts. GreensFirst vegetable supplement drink was tested to document its effect on the blood and saliva biochemistry in relation to cardiovascular health. RESULTS: After taking the supplement for 90 days, both the systolic and diastolic blood pressure decreased significantly. The heart rate decreased but did not reach statistical significance. Time domain analysis of HRV showed a slight decrease in standard deviation of normal-normal and square root of the mean squared differences but did not reach statistical significance. Frequency analysis of HRV found no significant changes. Saliva dehydroepiandosterone sulfate (DHEA-S) and Cortisol decreased after taking the supplement, however, they did not reach significant levels. Blood glucose concentration was increased slightly but the change was not significant. Blood serum tests showed no significant changes in the total cholesterol or the low density lipoprotein levels. However, it was noted that the high-density lipoprotein level was decreased significantly. Triglycerides did not show any significant changes but it were slightly elevated after the treatment period. Homocysteine increased significantly after taking the supplement for 90 days. CONCLUSION: For this group of subjects, taking the green vegetable drink for 90 days reduced blood pressure but did not significantly affect the blood and saliva chemistry. PMID- 19674673 TI - Is vitamin C an effective antihypertensive supplement? A review and analysis of the literature. AB - OBJECTIVE: Hypertension is a common condition with high mortality from associated diseases. Epidemiological evidence suggests that a dietary deficiency of vitamin C may be a risk factor for hypertension. However the literature on vitamin C interventional trials appears divided on the efficacy of vitamin C utilization clinically. METHODS: A literature search and review of published trials using vitamin C in treating patients with hypertension was undertaken. Relevant references were located using MEDLINE (1966-2005) and the bibliographies of located articles. RESULTS: Thirteen trials making up 14 separate groups were identified and analyzed providing a pooled population of 284 hypertensive patients (52% female), with a weighted mean age of 58.8 +/- 9.5 years. Median vitamin C dose and study intervention duration was 500mg/day and 6 weeks respectively. The weighted mean baseline and post treatment systolic blood pressures across all 14 groups were 149.6 +/- 11.1 and 145.7 +/- 11.0 mmHg respectively. This represented a systolic blood pressure decrease of 3.9 mmHg. Seven of the 14 groups ascertained statistically significant reductions (p < .05) in systolic blood pressures. However only 2 of the 14 groups found significant reductions in diastolic blood pressure. The weighted mean baseline and post treatment diastolic blood pressures across all 14 groups were 84.6 +/- 4.4 and 82.5 +/- 4.1 mmHg respectively. This represented a diastolic blood pressure decrease of 2.1 mmHg. CONCLUSION: Vitamin C supplementation in hypertensive patients appears to possess modest effects on reducing systolic blood and diastolic blood pressure. PMID- 19674674 TI - Chiropractic care of a six-year-old child with congenital torticollis. AB - OBJECTIVE: To discuss the care of a patient with congenital torticollis who was treated with chiropractic diversified technique and a chiropractic technique proposed by Frogley and Wallace in a previously published article. CLINICAL FEATURES: A 6-year-old female suffered from congenital torticollis with a left head-tilt subsequent to a difficult birth that culminated in a Caesarian section. Computerized tomography of the brain revealed plagiocephaly. Left facial asymmetry was demonstrated on visual observation. INTERVENTION AND OUTCOME: The patient was treated with a chiropractic adjusting protocol as suggested by Frogley and Wallace and by diversified chiropractic technique. She tolerated the care well and experienced substantial changes in clinical presentation including a distinct improvement in head-tilt. The improvement in head-tilt has been maintained for a period of 1 year. CONCLUSION: The conservative chiropractic methodology postulated by Frogley and Wallace in the care of headache patients should be investigated further in its use for congenial torticollis. The positive results observed with this patient lead the authors to recommend that additional studies be undertaken to assess whether these clinical observations can be replicated in other cases of congenital torticollis. PMID- 19674675 TI - Upper quarter kinetic chain response to cervical manipulation: a case report. AB - OBJECTIVE: The purpose of this single subject case study is to examine the relationship between cervical spine manipulation and tender points located in the upper quarter. CLINICAL FEATURES: A 45-year-old female presented with cervical spine pain and corresponding pain points in the upper quarter following a motor vehicle collision occurring 7 months prior to testing. Inclusion criteria were 22 of 22 clinically significant pain points as measured by pressure algometry. INTERVENTION AND OUTCOME: High velocity/low amplitude manual manipulation was administered to areas of joint restriction in the cervical spine. Pre- and post treatment computerized algometry readings were taken in areas of the cervical spine and upper extremities to assess change in pain over time. Algometry points were chosen based on published upper quarter tender points associated with cervical joint fixation/immobilization. CONCLUSION: Following a series of 15 cervical chiropractic manipulations, the patient demonstrated a significant increase in pain tolerance related to upper quarter pain points and demonstrated by pressure algometry. These findings correlate well with the literature, indicating that cervical spine joint dysfunction should be considered globally as part of a dynamic kinetic chain involving the upper quarter. Dysfunction in one part of the kinetic chain should direct treatment to other areas of the chain. PMID- 19674676 TI - How to write a case report for publication. AB - OBJECTIVE: This paper describes how and why to write a case report for publication in a peer-reviewed journal. METHODS: PubMed, the Cumulative Index to Nursing and Allied Health Literature (CINAHL), and the Index to Chiropractic Literature were searched from 2000 through September 2006 using the following search terms: case report, authorship, peer review, and manuscript. Relevant manuscripts were retrieved and the results were used to update a previous narrative overview of the literature. DISCUSSION: Commensurate with the increased use of evidence-based health care and recent changes in publication requirements, new standards are expected of case reports. Case reports should present new information to the literature and be written succinctly. The types of case reports available are discussed. Steps for preparing a case report are described based upon the current available literature. CONCLUSION: Case reports are important contributions to the health sciences literature. Proper preparation of this study design is necessary in order for it to be published. A self-evaluation check sheet for authors is included to assist in the writing process. PMID- 19674677 TI - The prevalence of positive imaging findings on MRI scans ordered by chiropractic versus medical providers. AB - OBJECTIVE: To determine if there is a greater yield of pathological findings identified on MRI scans of patients referred by chiropractors as compared to those referred by allopathic providers. METHODS: MRI reports authored by medical radiologists from two independent MRI centers in the Denver metropolitan area were analyzed retrospectively for pathological data related to the spinal regions studied. A pathological report data sheet was used to record pathological findings in 22 different categories. A total of 150 reports from each provider group were reviewed. RESULTS: Of the 22 pathological conditions studied, a statistically significant difference between doctor of chiropractic and medical doctor referrers was identified in 4 categories: central spinal canal stenosis, lateral stenosis, facet arthrosis, and negative report. The most common primary diagnoses given for MRI referral were low back pain/sciatica, neck pain, and extremity pain. Seventy-four percent of the reports evaluated were performed on patients referred with a diagnosis of pain. In 3 of the 22 categories (14%), the medical doctors had a statistically higher pathological yield than the chiropractors. However, in 4 of the 22 categories (18%), the chiropractors had a statistically higher pathological yield. In 18 of the 22 categories (82%), there was no statistical difference between the two provider groups. CONCLUSION: The data presented in this study suggests chiropractic and medical providers are compeer at ordering MRI for suspected pathological findings. PMID- 19674678 TI - A case series of reduced urinary incontinence in elderly patients following chiropractic manipulation. AB - OBJECTIVE: This report presents a spinal adjustment approach to treat elderly patients with urinary incontinence. METHODS: This retrospective case series reports the clinical observation of 13 patients with urinary incontinence. They were treated for 1-8 weeks with Pro-Adjuster technique without any other additional drug and physical therapy treatment. The primary outcome measure for this analysis was the frequency of nocturia reported by the patients during each office visit to the chiropractor. Bladder control was reported by the patients as frequency of nocturia or the urgency of urination or instances of leakage, and the number of pad changes per day or a return of the sense of urinary urge. RESULTS: A total of 13 patients' data (6 female, mean ages 65.7 +/- 12.9 years) were included in the study. After 1-8 weeks of chiropractic adjustments, the urinary frequency at night was significantly reduced from 3.8 to 1.2 time a night (P < 0.001). Three patients improved bladder control with only 2 adjustments. CONCLUSION: The Pro-adjuster treatment program seemed to reduce nocturia in patients with urinary incontinence. PMID- 19674679 TI - A manual therapy and exercise approach to meralgia paresthetica in pregnancy: a case report. AB - OBJECTIVE: To present a case of a pregnant patient with meralgia paresthetica who improved using manual therapy and exercise procedures. CLINICAL FEATURES: A 22 year-old patient in the sixteenth week of pregnancy had low back pain, bilateral anterolateral thigh paresthesia and groin pain for a duration of 1 month. She had no motor deficits in either lower extremity and her reflexes were intact. As a standard clinic procedure, a battery of functional tests were performed including: active straight leg raise, long dorsal ligament test, and the pelvic pain provocation procedure. Based on her clinical history and physical responses to the aforementioned functional tests, the diagnosis of meralgia paresthetica was deduced. INTERVENTION AND OUTCOME: Treatment was provided at 6 visits over a 6-week period where the patient underwent evaluation, manual intervention, and exercise prescription. Active Release Technique (ART) was performed to the restricted right sacroiliac (SIJ) complex and quadratus lumborum muscles. ART and post-isometric relaxation were applied to the illiopsoas muscles. The home exercise program consisted of pelvic/low back mobility, stabilization and relaxation exercises. After 6 treatments, the patient reported complete resolution of low back pain and left lower extremity symptoms and a 90% improvement in the right thigh symptoms. At her one-year follow-up, the patient reported no further complications and the absence of pain. CONCLUSIONS: Manual therapy and exercises may serve as an effective treatment protocol for pregnant patients experiencing low back pain complicated by paresthesia. Because these conservative procedures offer a low-risk intervention, additional clinical studies are warranted to further study this treatment. PMID- 19674680 TI - Use of conservative and sport-specific management strategies for a baseball pitcher with persistent elbow pain. AB - OBJECTIVE: To present a case of medial epicondylitis in an adolescent baseball pitcher that appeared to improve when addressing the upper extremity kinetic chain with a variety of conservative management strategies. CLINICAL FEATURES: An 18-year-old right hand dominant high school baseball pitcher had recurring sharp pain in his right elbow. Previous trials of medication and physical therapy were only partially successful in resolving the problem. The patient's pain was rated as an 8 on a numerical pain scale during throwing activities. A variety of tender points were noted in the right upper extremity. The patient had discontinued playing competitive baseball due to the pain. INTERVENTION AND OUTCOME: The patient was treated 14 times over 15 weeks. Treatment included cervical, thoracic and upper extremity manipulation, soft tissue mobilization and progressive sport specific training for the upper extremity. After the first treatment he was able to throw the ball and was able to continue throwing during the treatment program. The patient returned to league play after the treatment program. CONCLUSION: A course of conservative care addressing multiple elements of the upper extremity kinetic chain seemed beneficial for this athlete. PMID- 19674681 TI - Writing narrative literature reviews for peer-reviewed journals: secrets of the trade. AB - OBJECTIVE: To describe and discuss the process used to write a narrative review of the literature for publication in a peer-reviewed journal. Publication of narrative overviews of the literature should be standardized to increase their objectivity. BACKGROUND: In the past decade numerous changes in research methodology pertaining to reviews of the literature have occurred. These changes necessitate authors of review articles to be familiar with current standards in the publication process. METHODS: Narrative overview of the literature synthesizing the findings of literature retrieved from searches of computerized databases, hand searches, and authoritative texts. DISCUSSION: An overview of the use of three types of reviews of the literature is presented. Step by step instructions for how to conduct and write a narrative overview utilizing a 'best evidence synthesis' approach are discussed, starting with appropriate preparatory work and ending with how to create proper illustrations. Several resources for creating reviews of the literature are presented and a narrative overview critical appraisal worksheet is included. A bibliography of other useful reading is presented in an appendix. CONCLUSION: Narrative overviews can be a valuable contribution to the literature if prepared properly. New and experienced authors wishing to write a narrative overview should find this article useful in constructing such a paper and carrying out the research process. It is hoped that this article will stimulate scholarly dialog amongst colleagues about this research design and other complex literature review methods. PMID- 19674682 TI - The effects of active release technique on carpal tunnel patients: A pilot study. AB - OBJECTIVE: To examine changes in electromyography (EMG) and a valid self administered outcome measure after applying active release technique to carpal tunnel syndrome (CTS) patients. METHODS: Five subjects (mean age 48.2 SD +/- 16.7) with CTS were included in the trial. Subjects completed the Boston Questionnaire (BQ) and an EMG examination before the first treatment. Participants were treated with Active Release technique using a protocol intended to affect the median nerve 3 times a week for 2 weeks. The BQ was re-administered following the final treatment. The mean scores for the initial and final BQ were compared using a paired samples t-test. An analysis of variance compared the mean contraction amplitudes for EMG parameters before and after the first treatment. RESULTS: There was significant improvement (p < 0.05) in the mean symptom severity and functional status scores of the BQ following the intervention. There were no significant differences found in the EMG analyses. CONCLUSION: The preliminary data from this clinical pilot trial suggest that active release technique may be an effective conservative management strategy for CTS patients. These results support the need for further clinical trials with larger samples. PMID- 19674683 TI - Sagittal plane blockage of the foot, ankle and hallux and foot alignment prevalence and association with low back pain. AB - INTRODUCTION: Some practitioners believe mechanical low back pain may be caused or aggravated by a stiff ankle, stiff great toe or flat feet. This study investigates subjects with and without mechanical low back pain and measures ankle and great toe range of motion and flattening of the medial longitudinal arch in both groups. METHODS: The study was a blinded, 2-arm, non- randomized clinical study involving 100 subjects with chronic or recurrent mechanical low back pain (intervention group) and 104 subjects without chronic mechanical low back pain (control group) between the ages of 18 and 45. A blind assessor performed weight-bearing goniometry of the ankle and big toe and the navicular drop test on all subjects in both groups. RESULTS: An independent t-test (inter group) revealed a statistically significant decrease (p .05) in the control group. The total power reflecting the total autonomic activity was significantly decreased immediately after exercise and after the 20-minute rest period at the end of the exercise session in both the control and experimental groups. CONCLUSIONS: There were positive changes in cardiac and vascular autonomic regulations with exercise training when combined with foot orthotics. PMID- 19674696 TI - Intraexaminer and interexaminer reliability of mastoid fossa readings using a temporal artery thermometer. AB - OBJECTIVE: The temperature of the mastoid fossa region has been measured by chiropractors since the 1950s using various instruments. The reliability of the procedure is largely unknown for the various instruments used for this purpose. This study assessed the reliability of a thermal instrument designed to measure forehead temperature while having an alternate landmark as the mastoid fossa. METHODS: : Thirty students were recruited to participate. Three blinded examiners twice scanned each student with the temporal artery thermometer. Intraexaminer and interexaminer reliability was assessed using the intraclass correlation coefficient. Differential margins of error were also calculated. RESULTS: Intraexaminer reliability was acceptable in 6 of 6 assessments, whereas interexaminer reliability was acceptable in 3 of 4 assessments. The maximum mean differential margin of error was 0.54 for intraexaminer reliability and 0.46 for interexaminer reliability. DISCUSSION: Combining the mean maximum differential margin of error for intraexaminer (0.54 degrees ) and interexaminer (0.46 degrees ) and then averaging these 2 ([0.54 + 0.46] / 2) results in a maximum mean differential margin of error of 0.50 degrees . CONCLUSION: In this study, intraexaminer reliability ranged from fair to strong. For these examiners, fossa differentials of 0.5 can be considered within the margin of error. PMID- 19674697 TI - Pregnancy and chiropractic: a narrative review of the literature. AB - OBJECTIVE: The purpose of this article is to review the literature on the topic of chiropractic care during pregnancy. METHODS: A PubMed search was performed using the terms pregnancy and chiropractic. Sources were cross-referenced to obtain further articles and research information after reviewing the articles obtained through the search. RESULTS: Thirty-three references were used for this review. The current literature reports favorable results on the use of chiropractic care throughout pregnancy. CONCLUSIONS: Chiropractic evaluation and treatment during pregnancy may be considered a safe and effective means of treating common musculoskeletal symptoms that affect pregnant patients. The scarcity of published literature warrants further research. PMID- 19674698 TI - Physical assessment of lower extremity radiculopathy and sciatica. AB - OBJECTIVE: The purpose is to describe active and passive physical maneuvers that provoke or alleviate lumbosacral radiculopathy and/or sciatica to demonstrate how these maneuvers function in common orthopedic and neurological tests/signs. METHODS: Descriptions of the maneuvers and their countermaneuvers are provided to establish an understanding of their mechanisms and influence on lumbosacral nerve roots and the sciatic nerve. Common tests and signs for lower extremity radiculopathy and sciatica are described to demonstrate the clinical application of the maneuvers. Combinations of common tests are also offered to improve examination efficiency. RESULTS: Understanding how each maneuver contributes individually or in combination with standard tests will enhance the reader's examination skills. CONCLUSION: Improved examination skills lead to improved diagnosis and differential diagnosis of lower extremity radiculopathy and sciatica. PMID- 19674699 TI - Adolescent idiopathic scoliosis treatment using the Pettibon corrective procedures: a case report. PMID- 19674700 TI - The American Chiropractic Board of Sports Physicians supports the Journal of Chiropractic Medicine. PMID- 19674701 TI - Spinous process palpation using the scapular tip as a landmark vs a radiographic criterion standard. AB - OBJECTIVE: This study aimed at determining the standing spinal landmark that corresponds to the inferior tip of the scapula and determining the accuracy of experienced palpators in locating a spinous process (SP) 3 levels above and below a given SP. METHODS: The study participants were 34 asymptomatic or minimally symptomatic chiropractic students. An experienced palpator located the inferior scapular tip on each and then positioned a 2-mm lead marker about 5 cm lateral to the nearest SP. Two more markers were placed at levels intended to be 3 levels above and below the first marker placed. The locations of the scapular tip and the spinal targets were determined by comparison with a radiological criterion standard. RESULTS: The standing inferior scapular tip corresponded to the T8 SP on average (SD = 0.9). Having placed the first lead marker, examiners on average overshot the upper marker by 0.26 (SD = 0.51) vertebral levels and undershot the lower marker by 0.21 (SD = 0.48) vertebral levels. The modes for the placement of the 3 markers were at T5, T8, and T11. CONCLUSION: Approximately 68% of patients would be palpated to have their inferior scapular tips at T7, T8, or T9. An experienced palpator can quite accurately locate vertebral levels 3 above or below a given landmark. Chiropractors and other health professionals using the typical rule of thumb linking the inferior scapular tip to the standing T7 SP have likely been applying clinical interventions at spinal locations different from those intended. PMID- 19674702 TI - Triage and case presentations in a chiropractic pediatric clinic. AB - OBJECTIVE: The use of triage in a chiropractic practice is to determine whether or not a patient who has presented to an office is in need of a referral to another health care provider. The objective of this article is to illustrate the use of triage skills in a primary care, chiropractic pediatric practice. This is examined both in the new patient setting and during visit-to-visit protocol. METHOD: An analysis of the number of chiropractic triage visits during a 1-year period was performed on 48 new children or pregnant women and 1634 existing pediatric and pregnancy visits. RESULTS: The average level of chiropractic triage for a new patient, whether pediatric or pregnant, was a limited level of concern (P3, green flag), with more than 85% of new patients falling into that category. Fifteen percent were at a cautionary (P2, yellow flag) level, and none were at the emergency (P1, red flag) level. Between 12% and 15% of the total of existing pediatric or pregnancy visits were at a P1 or P2 level of triage, and 11% of the total of pediatric triage visits were at the P1 level. CONCLUSION: New patients rarely come to a chiropractic pediatric office in a P1-level crisis, whereas existing patients have a much higher likelihood of presenting during a P1 challenge. P2 triage levels are somewhat common, whereas P3 triage levels are the most frequent. Triage methods are a way to help place a patient in a category that can ensure an optimum, safe, and effective level of care. PMID- 19674703 TI - A review of the use of likelihood ratios in the chiropractic literature. AB - OBJECTIVE: The objective of this review is to determine how frequently likelihood ratios (LRs) have been used or described in the chiropractic literature and to depict their appropriate use in the clinical setting. METHODS: A literature search was conducted of the MEDLINE, Manual Alternative and Natural Therapy Index System, and Index to Chiropractic Literature databases, with search years encompassing 1966 through June 2006. Citations in the English language that addressed the following search terms were retrieved: likelihood ratio in combination with manipulation and chiropractic. RESULTS: The searches netted a total of 64 citations: 10 in MEDLINE, 34 in the Manual Alternative and Natural Therapy Index System, and 20 in the Index to Chiropractic Literature. After eliminating articles from journals that were not focused specifically on chiropractic, duplicates, and those that did not involve LRs, 3 articles remained and were reviewed. None of the reviewed articles provided a description of LRs, and only 2 used them in a clinical context. CONCLUSIONS: The use of LRs can be very helpful in patient management; however, LRs are rarely reported in the chiropractic literature. Accordingly, chiropractic practitioners are most likely uninformed on the subject and may not have the capacity to use them in formulating diagnoses. It is suggested that researchers increase the reporting of LRs and that chiropractic clinicians begin to make use of them in day-to-day practice. PMID- 19674704 TI - Correlation of health outcomes with physician and chiropractor ratios in the United States. AB - OBJECTIVE: There is controversy as to whether increased doctor-patient ratios result in a commensurate benefit to the health care consumer. This study assesses doctor (allopathic/osteopathic physician and chiropractor) ratios in the 50 states in the United States and correlates these ratios with various health outcomes to determine if one doctor type has stronger correlations in certain outcomes compared with the other doctor type by geographic region. METHODS: A total of 21 health outcomes for the year 2004, such as obesity and cancer deaths, were correlated with 2004 physician and chiropractor ratios (number of doctors per 100000 population). The 25 highest doctor ratio states, along with the corresponding health outcomes, were compared with the 25 lowest doctor ratio states and the corresponding health outcomes. The Spearman and Wilcoxon tests (for correlation and differences, respectively) were used to assess the data. RESULTS: Increases in doctor ratios resulted in correlations in 12 outcomes for chiropractors and 8 outcomes for allopathic/osteopathic physicians. When comparing low with high doctor ratios, physicians had improvements in 13 outcomes, whereas chiropractors had 12. CONCLUSIONS: Correlation does not necessarily show causation but may provide clues. Many of the improved outcomes were not surprising for allopathic/osteopathic physicians, for example, cardiovascular deaths, but were surprising for chiropractors. It is possible, although care should be taken to avoid overspeculation, that doctors of chiropractic are having an effect in seemingly unlikely outcomes such as cardiovascular and cancer deaths. Further research is warranted for other years to verify these findings. PMID- 19674705 TI - The etiology of cervical artery dissection. AB - The etiology of cervical artery dissection (CAD) is unclear, although a number of risk factors have been reported to be associated with the condition. On rare occasions, patients experience CAD after cervical spine manipulation, making knowledge about the cervical arteries, the predisposing factors, and the pathogenesis of the condition of interest to chiropractors. This commentary reports on the relevant anatomy of the cervical arteries, developmental features of CAD, epidemiology of the condition, and mechanisms of dissection. The analysis of CAD risk factors is confusing, however, because many people are exposed to mechanical events and known pathophysiological associations without ever experiencing dissection. No cause-and-effect relationship has been established between cervical spine manipulation and CAD, but it seems that cervical manipulation may be capable of triggering dissection in a susceptible patient or contributing to the evolution of an already existing CAD. Despite the many risk factors that have been proposed as possible causes of CAD, it is still unknown which of them actually predispose patients to CAD after cervical spine manipulation. PMID- 19674706 TI - Feasibility study of short-term effects of chiropractic manipulation on older adults with impaired balance. AB - OBJECTIVE: The purpose of this study was to collect preliminary information on the effects of chiropractic spinal manipulation on reducing risk of falls in older adults with impaired balance, as assessed by the Berg Balance Scale (BBS). This information is necessary to develop a line of investigation into the role of chiropractic care on reduction of fall risk in this population. METHODS: Randomized, 2-group pretest/posttest design feasibility study with a target sample size of 10 (5 per group), conducted within the outpatient health center of a chiropractic college. Inclusion criteria were as follows: aged 60 years or older, able to stand on one leg <5 seconds, and able to attend all sessions. Patients were assigned to chiropractic care (CMT) or supervised exercise (EX) and scheduled for 2 visits per week for 8 weeks. RESULTS: A total of 26 people responded to recruitment; and 11 were enrolled: 6 in the CMT and 5 in the EX group. Two patients dropped out at the baseline visit when they were assigned to the EX group. One CMT patient dropped out in the seventh week because of a fall at home resulting in a leg fracture. All remaining patients were compliant with treatment protocols. Five of 6 CMT patients and 4 of 5 EX patients had baseline BBS scores <45, indicating increased risk of falls. At visit 16, 2 CMT and 1 of the 3 remaining EX patients had BBS scores <45. One mild and transient adverse event was noted. CONCLUSION: Further investigation of the possible role of chiropractic care in reducing fall risk in this population appears feasible. PMID- 19674707 TI - Urinary incontinence in women treated by ischemic compression over the bladder area: a pilot study. AB - OBJECTIVE: The purpose of this study was to determine if ischemic compression therapy over the bladder area results in clinically important changes among female patients with stress and mixed (stress and urge) incontinence. METHODS: One group of patients (n = 24) received ischemic compression therapy directed over the bladder area (experimental group). The control group (n = 9) received ischemic compression therapy directed toward structures of the hip joint. Changes in urinary incontinence symptoms were monitored using a 2-part questionnaire: the urogenital distress inventory and the incontinence impact questionnaire. Patients' perceived amelioration (improvement) was quantified using a scale divided from 0% to 100%. RESULTS: Mean scores for the first questionnaire (urogenital distress inventory + incontinence impact questionnaire, 19 questions) were 23.3 vs 25.3 at baseline and 10.2 vs 22.2 after 15 treatments for the experimental and control group, respectively. The experimental group scores were 6.9 at 30 days after the last treatment and 11.3 at the 6-month follow-up. The perceived percentages of amelioration after 15 treatments were 69% vs 32% for the experimental and control group, respectively. The experimental group scores were 73% at 30 days after the last treatment and 60% at the 6-month follow-up. CONCLUSIONS: In this study, ischemic compression directed toward elicited trigger points over bladder area was found to be an effective treatment of patients presenting symptoms of urinary incontinence. Improvement in symptoms was still present in follow-up at 6 months. PMID- 19674708 TI - The effect of cervical spine manual therapy on normal mouth opening in asymptomatic subjects. AB - OBJECTIVE: Temporomandibular disorders are a group of disorders affecting the temporomandibular joint and/or masticatory muscles. One of the signs associated with temporomandibular disorders is a reduction in mouth opening. During normal mouth opening, extension occurs at the cervical-cranial junction. The purpose of this investigation was to determine if manual therapy applied to the cervical cranial junction would significantly improve mouth-opening capacity. METHODS: One hundred one participants were randomly assigned to either an Active Release Technique (ART) group; high-velocity, low-amplitude manipulation (HVLA) group; or control group. A blinded investigator measured mouth opening using a TheraBite range of motion scale (TheraBite Corporation, West Chester, PA). Participants received ART to the suboccipitals or HVLA to the cervical spine at C1 or sat with an investigator for 3 minutes with no treatment. After the treatment session, mouth opening was remeasured. A repeated-measures analysis of variance was used to compare the group mean values. The a priori alpha level was .05. RESULTS: The repeated-measures analysis of variance showed no significant difference between the ART, HVLA, and control groups' pretreatment and posttreatment measurements (F = 0.41, P > .05). CONCLUSION: Manual therapy to the cervical spine did not significantly improve mouth opening in this asymptomatic population. Future trials using participants with restricted mouth-opening measures are warranted. PMID- 19674709 TI - Electromyographic responses from the stimulation of the temporalis muscle through facial acupuncture points. AB - OBJECTIVE: This study measured the electromyographic (EMG) responses of the temporalis muscle to electrical stimulation at 10 acupuncture points (acupoints) of the face and assessed whether electrical stimulation of these acupoints may improve the rhythmic activity of the muscles that open and close the jaw. METHODS: Fifty healthy adults clenched 2 thin pieces of wood between their teeth and kept a constant bite force for 30 seconds. The left anterior temporalis muscle was chosen for spontaneous EMG stimulation and recording. RESULTS: The mean EMG responses were positive at 10 points in early reflexes but negative at points Jiache, Chengjiang, and Dicang for late reflexes. No statistically significant difference was observed in the responses between age and stimulation intensity for both areas, but a significant inverse correlation existed between the distance from the acupoint to the recording electrode and the response. CONCLUSION: This study showed that the surface electrical current applied to facial skin acupoints may excite or inhibit the anterior temporalis muscle via reflex pathways. The result from this stimulation may be applied to treat temporomandibular joint disorder occlusion of muscular origin. PMID- 19674711 TI - Undiagnosed Brodie abscess in a gymnast after surgical fixation of a tibial fracture. AB - OBJECTIVE: This study presents a case of a posttraumatic subacute osteomyelitis in a child with leg pain. CLINICAL FEATURES: A 10-year-old female gymnast with leg pain presented to a chiropractic clinic after having been treated over the previous year for a leg fracture. The patient had leg pain associated with prolonged use of her right leg, restlessness at night, and tenderness over the right tibia. The history did not suggest a mechanical cause of the patient's pain. All available radiographs were reviewed by the chiropractor; a diffuse lytic lesion with bone thickening and sclerosis was clearly visible in the area of the patient's chief complaint, representing a Brodie abscess. INTERVENTION AND OUTCOME: The doctor of chiropractic sent the patient back to the hospital. She was treated first with oral antibiotics, which were not successful. She underwent surgery and recovered well. CONCLUSION: Subacute osteomyelitis may have a diagnostic delay; thus, it is possible for a chiropractor to see this condition in the office. A good case history, examination, and radiographs are important for the diagnosis and to make a proper referral. PMID- 19674710 TI - Chiropractic treatment of a pregnant patient with lumbar radiculopathy. AB - OBJECTIVE: The purpose of this report is to describe chiropractic treatment of lower back and unilateral leg pain in a pregnant patient. CLINICAL FEATURES: A 26 year-old woman in her second trimester of pregnancy had severe pain in her lower back that radiated to her hips bilaterally and to her right leg. She reported tingling down her right lower leg to the dorsum of her foot. Although no diagnostic imaging was performed, her differential diagnoses included lumbalgia with associated radiculopathy. INTERVENTION AND OUTCOME: Treatment consisted of manual traction in the side-lying position using a specialized chiropractic table and treatment technique (Cox flexion-distraction decompression) modified for pregnancy. Relief was noted after the first treatment, and complete resolution of her subjective and objective findings occurred after 8 visits. CONCLUSION: When modified, this chiropractic technique appears to be an effective method for treating lower back pain with radiation to the leg in a pregnant patient who cannot lie prone. PMID- 19674712 TI - Combination testing in orthopedic and neurologic physical examination: a proposed model. AB - OBJECTIVE: This article suggests a 4-part model for teaching and using orthopedic and neurologic physical testing. DISCUSSION: Four methods of combining and sequencing orthopedic and neurologic physical tests are described. The descriptions are followed by examples including test names, test performance, and the relationships between the tests in each group. The principles of the methods originated in the lead author's private practice and were refined while teaching chiropractic students and graduate doctors. CONCLUSION: This model offers one possible method of combining and sequencing the orthopedic and neurologic examination in an effort to provide a more complete picture portraying the mechanisms, results, pathologies, differential diagnosis, and clinical thought processes associated with common orthopedic and neurologic physical tests. PMID- 19674713 TI - A feasibility study assessing manual therapies to different regions of the spine for patients with subacute or chronic neck pain. AB - OBJECTIVE: The purpose of this project was to develop and test protocols for a randomized clinical trial of a combined therapeutic approach (thoracic spine and sacroiliac joint high-velocity, low-amplitude spinal manipulation [HVLA SM] + cervical spine postisometric relaxation) and cervical spine HVLA SM for patients with subacute or chronic neck pain. METHODS: Patients were recruited in the Quad Cities in Iowa and Illinois. After a baseline assessment visit, eligible patients were randomly assigned to cervical spine HVLA SM or to the combined therapeutic approach for 4 treatment visits over 2 weeks. Outcome assessments included the Neck Disability Index, visual analog scale, and posttreatment response questionnaire. Patient outcomes were not aggregated or compared by treatment group. RESULTS: It took approximately 8 months of planning, which included the development of forms and protocols, pretesting the forms, and training staff and clinicians in the standardized protocols. Twelve participants were screened, and 6 patients were enrolled and randomly allocated to care over a 6-week period. All patients completed 5 visits. Five of 6 patients had an improvement on the Neck Disability Index. On the visual analog scale, 2 patients improved at 2 weeks, whereas the other 4 got worse. Five patients completed the posttreatment response questionnaire; 2 of the 5 indicated they experienced discomfort or an unpleasant reaction from the study treatments. CONCLUSIONS: Designing a successful feasibility randomized clinical trial requires considerable planning, development and pretesting of the forms and protocols, and training clinicians and staff for standardized protocols. Patients were willing to be randomized, follow treatment protocols, complete baseline and outcome assessments, and return 83% of the follow-up questionnaires. PMID- 19674714 TI - Surface area congruence of atlas superior articulating facets and occipital condyles. AB - OBJECTIVE: The purpose of this study was to compare the surface areas of 12 atlanto-occipital joints from 6 cadavers to determine how well their ipsilateral and contralateral surface areas matched. METHODS: Three methods were used. Method 1 consisted of digitized photographs downloaded to the Able Image Analyser software program (Ljubljana, Slovenia; http://able.mulabs.com). The perimeters were measured and expressed as square millimeters. Method 2 consisted of a point count method using moulds of the joint surfaces produced by pressing aluminum paper, leaving a clear imprint for analysis. Method 3 consisted of drawing outlines of the molds from method 2 onto transparencies and assessing overlap. RESULTS: Method 1 showed a moderate correlation of matched articular surfaces between the left C1 superior articulating surface and the left condyle (r = 0.573, P = .01). Method 2 showed moderate correlations between all surface areas that were analyzed. The matched pairs compared were left C1 superior articulating surface and left condyle (r = 0.588, P = .01) and right C1 and right condyle (r = 0.730, P = .001). The contralateral surfaces correlated were left C1 and right C1 (r = 0.596, P = .009) and right condyle and left condyle (r = 0.769, P = .000). Method 3 showed no statistically significant differences between surface areas. CONCLUSIONS: All 3 methods revealed that the articular surfaces of the atlas and corresponding or contralateral condyle for specimens used in this study were not an exact match. PMID- 19674715 TI - Resolution of cervical radiculopathy in a woman after chiropractic manipulation. AB - OBJECTIVE: To describe a case regarding a woman with 2-level cervical disk herniation with radicular symptoms conservatively treated with chiropractic care including high-velocity, low-amplitude (HVLA) manipulation with complete resolution of her symptoms. CLINICAL FEATURES: A 40-year-old woman developed right finger paresthesia and neck pain. Results of electrodiagnostics were normal, but clinical examination revealed subtle findings of cervical radiculopathy. A subsequent magnetic resonance imaging revealed a large right posterolateral disk protrusion and spur impinging on the right hemicord with moderate to severe central canal and right neuroforaminal stenosis at C5-6 and C6 7. She was treated with HVLA manipulation to the cervical spine, as well as soft tissue techniques, traction, nonsteroidal anti-inflammatory drugs, and exercise. INTERVENTION AND OUTCOME: Her clinical findings and symptoms resolved within 90 days of initiating care and did not return in 1 year. There were no untoward effects, including transient ones. CONCLUSION: This case describes the clinical presentation and course of a patient with multilevel large herniated disks and associated radiculopathy who was treated with HVLA manipulation and other conservative approaches and appeared to have good outcomes. PMID- 19674716 TI - A case study of back pain and renal cell carcinoma. AB - OBJECTIVE: Renal cell carcinoma accounts for 3% of adult malignancy and 95% of neoplasms arising from the kidney. One third of the patients have metastatic disease at the time of presentation and are asymptomatic with the diagnosis being made incidentally from a radiologic study obtained for other reasons. Typically, skeletal metastases are purely lytic. It is common for cases of back pain to be managed in chiropractic clinics. The diagnosis of metastatic disease is to be considered as a differential, especially in an aging population. CLINICAL FEATURES: A 69-year-old man experienced back pain after a twisting movement while working. He presented for chiropractic care 2 weeks later. He had a 10-year history of low back pain and a 2-month history of weakness at the knee. Plain film radiography was assessed as negative for pathology by a medical radiologist and a chiropractic radiologist. INTERVENTIONS AND OUTCOMES: After 6 visits of care using atlas orthogonal adjusting, the outcome assessment revealed improvement in the pain but a regression in strength of the right quadriceps and peroneus muscles. The patient was referred for a magnetic resonance imaging study, which revealed a mass on the kidney and a large erosive bone lesion of the L4 vertebrae. The diagnosis of renal cell carcinoma was later confirmed. CONCLUSION: Progressive muscle weakness is one of the "red flag" signs in a back pain case. Advanced imaging is necessary to detect occult diseases processes such as renal cell carcinoma. The chiropractor's role as primary care clinician includes the process of clinical reasoning and appropriate referral when the clinical picture becomes unclear and/or "red flag" signs appear. PMID- 19674717 TI - Slow progressing cardiac complications-a case report. AB - OBJECTIVE: This case presentation describes an uncommon development of complete heart block. Within 48 hours after a motor vehicle accident with the deployment of the air bag against the patient's chest, the patient reported exertional bradycardia and shortness of breath. CLINICAL FEATURES: A 51-year-old man was in a motor vehicle accident. After the collision, he noticed a slow onset of chest discomfort with exertion and bradycardia. The patient experienced cardiac difficulty during a stress electrocardiogram. During the 4 months after the motor vehicle accident, symptoms progressed; and a diagnosis of vagal sympathetic reflex was suggested. INTERVENTION AND OUTCOME: A pacemaker was finally required because of the ventricular pacing of 35 to 40 beats per minute, which was symptomatic of a complete atrioventricular block. CONCLUSION: A gradual progression to complete atrioventricular block over a period longer than 3 weeks is unusual. This case demonstrates that a patient manifesting exertional bradycardia and shortness of breath shortly after chest trauma should be regularly monitored until all symptoms are resolved. PMID- 19674718 TI - Cloth-covered chiropractic treatment tables as a source of allergens and pathogenic microbes. AB - OBJECTIVE: Vinyl chiropractic tables have been found to harbor pathogenic bacteria, but wiping with a simple disinfection agent can significantly reduce the risk of bacteria. The aim of this study was to assess the presence of microbes and other allergens or pathogens on cloth chiropractic tables. METHODS: Cloth-covered tables in a chiropractic college teaching clinic were selected. Samples were taken from the facial piece and hand rests with RODAC plates containing nutrient agar, followed by confirmatory testing when indicated. RESULTS: Numerous microbacteria strains were found, including Staphylococcus aureus and Propionibacterium. Allergen-producing molds, including Candida, were also found. CONCLUSION: Cloth tables were shown to contain pathogenic microbacteria and allergens. The chiropractic profession should establish an infection control protocol relevant to treatment tables and discard use of cloth covered treatment tables in this process. PMID- 19674719 TI - Effect of chiropractic treatment on hip extension ability and running velocity among young male running athletes. AB - OBJECTIVE: This study investigates the effect of chiropractic treatment on hip joint extension ability and running velocity. METHODS: This was a prospective, randomized, controlled experimental pilot study. Seventeen healthy male junior athletes (age, 17-20 years) training in middle distance running were recruited from local Swedish athletic associations. Hip extension ability and running velocity were measured before and after the study period. Chiropractic investigations comprised motion palpation of the sacroiliac and hip joints and modified Thomas test of the ability to extend the leg. In the treatment group, findings of restrictive joint dysfunctions formed the basis for the choice of chiropractic treatment. The interventions were based on a pragmatic approach consisting of high-velocity, low-amplitude manipulations targeted toward, but not exclusively to, the sacroiliac joints. RESULTS: The treatment group showed significantly greater hip extension ability after chiropractic treatment than did controls (P < .05). Participants in the treatment group did not show a significant decrease in time for running 30 m after treatment (average, -0.065 seconds; P = .0572), whereas the difference was even smaller for the control subjects (average, -0.003; P = .7344). CONCLUSIONS: The results imply that chiropractic treatment can improve hip extensibility in subjects with restriction as measured by the modified Thomas test. It could be speculated that the running step was amplified by increasing the angle of step through facilitated hip joint extension ability. The possible effect of chiropractic treatment to enhance the running velocity, by increasing the hip joint extension ability and thereby increasing the running step, remains unproven. PMID- 19674720 TI - Vitamin C supplementation lowers serum low-density lipoprotein cholesterol and triglycerides: a meta-analysis of 13 randomized controlled trials. AB - OBJECTIVE: Vitamin C has been shown to be an effective therapeutic for reducing total serum cholesterol, but epidemiologic studies have determined that low density lipoprotein (LDL) cholesterol and high-density lipoprotein (HDL) cholesterol are actually better predictive measures of coronary heart disease risk. Therefore, the purpose of this study was to provide a comprehensive meta analysis of randomized controlled trials to investigate the effect of vitamin C supplementation on LDL and HDL cholesterol as well as triglycerides in patients with hypercholesterolemia. METHODS: Thirteen randomized controlled trials published between 1970 and June 2007 were identified using Medline and a manual search. From the 13 trials, 14 separate group populations with hypercholesterolemia and who were supplemented with at least 500 mg/d of vitamin C for between 3 and 24 weeks were entered into the meta-analysis. This meta analysis used a random-effects model; and the overall effect sizes were calculated for changes in LDL and HDL cholesterol, as well as triglyceride concentrations. RESULTS: The pooled estimate of effect for vitamin C supplementation on LDL and HDL cholesterol was -7.9 mg/dL (95% confidence interval [CI], -12.3 to -3.5; P = .000) and 1.1 mg/dL (95% CI, -0.2 to 2.3; not significant), respectively. The pooled estimate of effect for vitamin C supplementation on triglycerides was -20.1 mg/dL (95% CI, -33.3 to -6.8; P < .003). CONCLUSION: Supplementation with at least 500 mg/d of vitamin C, for a minimum of 4 weeks, can result in a significant decrease in serum LDL cholesterol and triglyceride concentrations. However, there was a nonsignificant elevation of serum HDL cholesterol. PMID- 19674721 TI - Effects of Biofreeze and chiropractic adjustments on acute low back pain: a pilot study. AB - OBJECTIVE: This randomized controlled study was designed to determine the pain relieving effect of Biofreeze (Performance Health Inc., Export, PA) body surface application and chiropractic adjustments on subjects with acute low back pain (LBP). METHODS: The data were collected at the baseline, 2 weeks after treatment, and 4 weeks after treatment for final analyses. Diversified manual adjustments were provided by licensed chiropractors twice a week for 4 weeks to both control and experimental groups. Biofreeze was applied to the lower back area 3 times a day for 4 weeks in the experimental group. Outcome assessments included visual analog scale, Roland Morris Disability Questionnaire, heart rate variability for stress, and electromyography for low back muscle activity. RESULTS: A total of 36 subjects were recruited in the study (25 male). The average age was 34 years. Significant pain reduction was found after each week of treatment in the experimental group (P < .05). The Roland Morris Disability Questionnaire did not show significant changes in both groups. There were no significant differences for pain reduction in the control group. Heart rate variability analysis showed no significant change (P > .05) in the experimental group after 4 weeks of Biofreeze and chiropractic adjustments. There were no statistically significant changes in the electromyography readings between the 2 groups. CONCLUSION: Biofreeze combined with chiropractic adjustment showed significant reduction in LBP. PMID- 19674722 TI - A case of chronic migraine remission after chiropractic care. AB - OBJECTIVE: To present a case study of migraine sufferer who had a dramatic improvement after chiropractic spinal manipulative therapy (CSMT). CLINICAL FEATURES: The case presented is a 72-year-old woman with a 60-year history of migraine headaches, which included nausea, vomiting, photophobia, and phonophobia. INTERVENTION AND OUTCOME: The average frequency of migraine episodes before treatment was 1 to 2 per week, including nausea, vomiting, photophobia, and phonophobia; and the average duration of each episode was 1 to 3 days. The patient was treated with CSMT. She reported all episodes being eliminated after CSMT. The patient was certain there had been no other lifestyle changes that could have contributed to her improvement. She also noted that the use of her medication was reduced by 100%. A 7-year follow-up revealed that the person had still not had a single migraine episode in this period. CONCLUSION: This case highlights that a subgroup of migraine patients may respond favorably to CSMT. While a case study does not represent significant scientific evidence, in context with other studies conducted, this study suggests that a trial of CSMT should be considered for chronic, nonresponsive migraine headache, especially if migraine patients are nonresponsive to pharmaceuticals or prefer to use other treatment methods. PMID- 19674723 TI - Public health advocacy and chiropractic: a guide to helping your community reach its health objectives. AB - OBJECTIVE: Doctors of chiropractic (DCs) provide health educational and promotion efforts in the communities they serve by counseling patients at the individual level. This article outlines a method and model in which DCs can effectively serve as public health advocates within their community. DISCUSSION: The social ecological model of health education and health promotion serves as an excellent template for taking into account every antecedent to disease within a community and how to prevent it through health promotion. A step-by-step guide to getting the DC involved in the community can be centered on this model, with the DC serving as a health advocate for his or her community. Resources are provided to assist in this process. CONCLUSION: The DC can and should engage his or her community in areas that are conducive to health through involvement and advocacy roles where these are suitable. A community's health can be enhanced with greater health care provider involvement, and DCs need to consider themselves a part of this process. PMID- 19674724 TI - Maximum vs. minimum values: preferences of speakers and listeners for upper and lower limit estimates. AB - Estimates about uncertain quantities can be expressed in terms of lower limits (more than X, minimum X), or upper limits (less than Y, maximum Y). It has been shown that lower limit statements generally occur much more often than upper limit statements (Halberg & Teigen, 2009). However, in a conversational context, preferences for upper and lower limit statements will be moderated by the concerns of the interlocutors. We report three studies asking speakers and listeners about their preferences for lower and upper limit statements, in the domains of distances, durations, and prices. It appears that travellers prefer information about maximum distances and maximum durations, and buyers (but not sellers) prefer to be told about maximum prices and maximum delivery times. Mistaken maxima are at the same time regarded as more "wrong" than mistaken minima. However, this preference for "worst case" information is not necessarily shared by providers of information (advisors), who are also concerned about being blamed if wrong. PMID- 19674725 TI - Peripheral branching of levator superioris muscle and Muller muscle origin. AB - PURPOSE: To examine the structure of the distal levator superioris muscle and the origin of the Muller muscle. DESIGN: Experimental dissectional study. METHODS: Specimens from 10 postmortem upper eyelids and orbits of 7 white cadavers (6 right eyes and 4 left eyes; age range, 78 to 101 years of age at death; age average, 87.7 years) were used for microscopic observations. The upper eyelids and orbits with sagittal full-thickness sections of the central part were examined microscopically using Masson trichrome staining. RESULTS: Sections from 9 of the specimens were suitable for analysis. The levator superioris muscle divided into a superior and an inferior branch by connective tissue in the peripheral region in all 9 samples. Although the thickness of each branch varied, that of the superior branch tended to be thicker than that of the inferior branch. In 8 specimens of 7 cadavers, the Muller muscle originated from the distal end of the inferior branch of the levator superioris muscle. In one specimen, the Muller muscle originated from the distal end as well as the posterior aspect of the levator superioris muscle. The levator aponeurosis originated from the superior branch. CONCLUSIONS: The distal levator superioris muscle consisted of a branched structure, and the Muller muscle originated from the inferior branch of the levator superioris muscle. PMID- 19674726 TI - Failed descemet-stripping automated endothelial keratoplasty grafts: a clinicopathologic analysis. AB - PURPOSE: To describe the clinicopathologic findings in failed Descemet-stripping automated endothelial keratoplasty (DSAEK) grafts. DESIGN: Retrospective, interventional case series. METHODS: SETTING: New York Eye and Ear Infirmary. STUDY POPULATION: Twenty-one patients with 22 failed DSAEK grafts treated between March 1, 2006 and February 1, 2008. INTERVENTION: Repeat DSAEK or penetrating keratoplasty were performed in the eyes with failed grafts. All failed grafts were examined histopathologically. MAIN OUTCOME MEASURES: Histopathologic parameters studied in failed DSAEK grafts included endothelial cell count, interface characteristics, retrocorneal membrane formation, inflammation, and immunoreactivity for herpes simplex virus type 1 (HSV-1) antigen. RESULTS: DSAEK failure was strongly associated with postoperative lenticle dislocation. Graft failure was primary in 19 DSAEKs and secondary to rejection, eccentric trephination with epithelial ingrowth, or bacterial infection in the remaining 3. All failed grafts demonstrated endothelial hypocellularity and stromal edema. Additional findings included stromal inflammation (68%), interface fibrosis (50%), retrocorneal membrane (36%), unplanned retention of Descemet membrane (14%), immunoreactivity for HSV-1 (14%), paucicellular stroma (14%), and uneven trephination with epithelial ingrowth (5%). CONCLUSIONS: Most DSAEK failures are secondary to endothelial cell loss. Other contributing factors include interface fibrosis, retrocorneal membrane formation, retained host Descemet membrane, uneven trephination, epithelial ingrowth, graft rejection, and infection. PMID- 19674728 TI - Chronic herpes simplex scleritis: characterization of 9 cases of an underrecognized clinical entity. AB - PURPOSE: To characterize in detail persistent and recalcitrant infectious scleritis resulting from herpes simplex virus (HSV). DESIGN: Retrospective and interventional clinical and immunopathologic case series. METHODS: Nine patients with chronic scleral redness, edema, and pain refractory to conventional therapy underwent a scleroconjunctival biopsy for routine histopathologic evaluation and definitive immunodiagnosis for the presence of HSV. Immunofluorescent probing for HSV was performed on the patient specimens. Negative controls for HSV included elimination of anti-HSV and anti-varicella zoster virus antibody in the probing process and the use of normal human conjunctiva and sclera as substrates. Response to therapy with acyclovir was monitored and recorded. RESULTS: The average age of the affected patients was 50.2 years, and the average duration of symptoms before tissue diagnosis of herpetic scleritis was 3.2 years (median, 4 years). Three histopathologic patterns were discovered: granulomatous inflammation (2 cases), plasma cell-rich pyogenic granuloma-like pattern (1 case), and reactive fibroinflammatory pattern (6 cases). Herpes antigen was demonstrated uniformly by immunofluorescence in a perivascular distribution and less commonly in the interstitium. Varicella zoster virus was not detected, and all controls for nonspecific antibody reagent binding to tissue showed negative results. Acyclovir caused a dramatic improvement in the chronic or recurrent ocular inflammation in all instances, with an average duration of improvement of inflammation of 15.3 months. CONCLUSIONS: Unrecognized HSV infection can cause longstanding scleritis. Histopathologic features of HSV scleritis are varied and nonspecific; immunofluorescent demonstration of HSV protein can make a definitive diagnosis. Prolonged administration of acyclovir is required for effective therapy. PMID- 19674730 TI - Confocal microscopy in ophthalmology. AB - PURPOSE: To describe the principles, capabilities, and applications of confocal microscopy in vivo in ophthalmology. DESIGN: Perspective, literature review, and commentary. METHODS: Review and synthesis of selected recent literature, with interpretation and perspective. RESULTS: Confocal microscopy imaging has led to a better understanding of the cellular microstructure in the normal, postsurgical, and diseased cornea by enabling quantitative analysis of the cellular response in the human cornea in vivo. At present, the major role of confocal microscopy is in research of corneal surgery and disease. Clinical applications are limited to facilitating the diagnosis of Acanthamoeba and deep fungal keratitis, measuring residual bed thickness after laser in situ keratomileusis, and measuring endothelial cell density in high-light-scattering situations. CONCLUSIONS: In addition to providing qualitative data, confocal microscopy is valuable for quantitative analysis of the cornea and will enable the investigation of pharmacologic and surgical modifications of corneal wound healing, nerve regeneration, and cellular responses. Prospective, quantitative analyses require individual calibration of confocal microscopes for lateral and axial dimensions of images, for image depth, and for light intensity. PMID- 19674729 TI - Three-year follow-up of the tube versus trabeculectomy study. AB - PURPOSE: To report 3-year results of the Tube Versus Trabeculectomy (TVT) Study. DESIGN: Multicenter randomized clinical trial. METHODS: SETTING: Seventeen clinical centers. STUDY POPULATION: Patients 18 to 85 years of age who had previous trabeculectomy, cataract extraction with intraocular lens implantation, or both and uncontrolled glaucoma with intraocular pressure (IOP) > or =18 mm Hg and < or =40 mm Hg on maximum tolerated medical therapy. INTERVENTIONS: A 350 mm(2) Baerveldt glaucoma implant or trabeculectomy with mitomycin C (MMC 0.4 mg/ml for 4 minutes). MAIN OUTCOME MEASURES: IOP, visual acuity, use of supplemental medical therapy, surgical complications, and failure (IOP >21 mm Hg or not reduced by 20%, IOP < or =5 mm Hg, reoperation for glaucoma, or loss of light perception vision). RESULTS: A total of 212 eyes of 212 patients were enrolled, including 107 in the tube group and 105 in the trabeculectomy group. At 3 years, IOP (mean +/- standard deviation [SD]) was 13.0 +/- 4.9 mm Hg in the tube group and 13.3 +/- 6.8 mm Hg in the trabeculectomy group (P = .78). The number of glaucoma medications (mean +/- SD) was 1.3 +/- 1.3 in the tube group and 1.0 +/- 1.5 in the trabeculectomy group (P = .30). The cumulative probability of failure during the first 3 years of follow-up was 15.1% in the tube group and 30.7% in the trabeculectomy group (P = .010; hazards ratio, 2.2; 95% confidence interval, 1.2 to 4.1). Postoperative complications developed in 42 patients (39%) in the tube group and 63 patients (60%) in the trabeculectomy group (P = .004). Surgical complications were associated with reoperation and/or loss of > or =2 Snellen lines in 24 patients (22%) in the tube group and 28 patients (27%) in the trabeculectomy group (P = .58). CONCLUSIONS: Tube shunt surgery had a higher success rate compared to trabeculectomy with MMC during the first 3 years of follow-up in the TVT Study. Both procedures were associated with similar IOP reduction and use of supplemental medical therapy at 3 years. While the incidence of postoperative complications was higher following trabeculectomy with MMC relative to tube shunt surgery, most complications were transient and self limited. PMID- 19674731 TI - Changes in aqueous vascular endothelial growth factor and pigment epithelium derived factor after ranibizumab alone or combined with verteporfin for exudative age-related macular degeneration. AB - PURPOSE: To investigate changes in aqueous vascular endothelial growth factor (VEGF) and pigment epithelium-derived factor (PEDF) in choroidal neovascularization (CNV) secondary to age-related macular degeneration (AMD) after ranibizumab (Lucentis; Genentech Inc, South San Francisco, California, USA) monotherapy or combined with photodynamic therapy (PDT). DESIGN: Prospective, interventional, case-control study. METHODS: We recruited 34 patients with CNV secondary to AMD and 10 controls. Baseline examinations, including visual acuity (VA), central macular thickness (CMT), fluorescein angiography, and indocyanine angiography, were performed, and the measurements of VA and CMT were repeated 1 month after treatments. Seventeen of 34 patients received a single intravitreal injection of 0.5 mg ranibizumab, and the remaining 17 patients underwent combined PDT on the same day. Aqueous samples were collected at the time of injection and 1 month after treatment and were measured by enzyme-linked immunosorbent assay. Main outcomes measures were the changes in VA and CMT and the changes in VEGF and PEDF levels. RESULTS: Demographic features, lesion characteristics, and mean changes in VA and CMT were similar between the two groups. Aqueous VEGF and PEDF levels were reduced significantly 1 month after treatment in all patients. The reduction levels of VEGF and PEDF were similar between the two groups. There was a positive correlation between the reduction levels of aqueous VEGF and the reduction levels of aqueous PEDF. The reduction levels of VEGF and PEDF were correlated positively with the decrease in CMT, but were not positively correlated with the improvements in VA. CONCLUSIONS: Ranibizumab therapy for CNV secondary to AMD is associated with reduced levels of aqueous VEGF and PEDF regardless of combined therapy with PDT. The reduction levels of VEGF and PEDF are correlated with anatomic improvements in the macula. PMID- 19674732 TI - Prevalence and associations of keratoconus in rural maharashtra in central India: the central India eye and medical study. AB - PURPOSE: To evaluate the prevalence and associated factors of keratoconus in the adult Indian population. DESIGN: Population-based study. METHODS: The Central India Eye and Medical Study is a population-based study performed in a rural region close to Nagpur in Central India. It included 4,711 subjects (aged 30 years and above) out of 5,885 eligible subjects (response rate, 80.1%). The participants underwent a detailed ophthalmic and medical examination. This study was focused on the prevalence of keratoconus, defined as an anterior corneal refractive power exceeding 48 diopters (D). RESULTS: Out of the 4,711 subjects, corneal refractive power measurements were available for 4,667 subjects (99.1%). Keratoconus was detected in 212 eyes (2.3%) of 128 subjects (prevalence rate, 2.3% +/- 0.2%). In multivariate analysis, the presence of keratoconus was significantly associated with lower body height (P < .001), lower level of education (P= .03), higher myopic refractive error (P = .004), and thinner central corneal thickness (P = .006). It was not significantly associated with alcohol consumption (P = .99) or smoking (P = .08) nor with questions relating to the psychiatric status. Defining a keratoconus as corneal refractive power of > or =49 D or of > or =50 D, a keratoconus was detected in 58 eyes (0.6%) and 10 eyes (0.1%), respectively. CONCLUSIONS: Keratoconus defined as corneal refractive power of 48+ D has a prevalence of 2.3% +/- 0.2% among Indians aged 30 years and above and living in the rural region of Central India. The prevalence of keratoconus was associated with lower body height, lower level of education, myopic refractive error, and thinner central cornea. PMID- 19674733 TI - Injury severity assessment for underground coalmine workers. AB - An injury severity model is proposed for assessment of injury incidents in industrial settings. A classification scheme for injury incidents considering interactions is also developed. The injury severity model considers injury potential in the form of unsafe conditions and analyzes its transfer to actual injury of varying severity. A case study was conducted in an underground coalmine of eastern India. An observed reduction in risk realization is explained through the model. Presence of interactions is found to be the most significant incident attribute of injury occurrences. The classification scheme and the results obtained from this study will help in improving accident/injury investigation reporting and devising preventive measures for reducing injury severity. PMID- 19674735 TI - Sorting and disposing of waste at recycling centres--a user's perspective. AB - This paper investigates Swedish recycling centres from the users' perspective. The aim was to describe the characteristics and experiences of the users and their activities when sorting and disposing of waste, and to identify improvements for the users. The typical recycling centre user is a recently retired man, living in a house with a garden, having travelled 5 km alone in his own car. The users requested longer opening hours and better information available at home and at the recycling centre. The major difficulty for the users is to understand which fraction their waste belongs to, and consequently into which container they should throw it. The most important sources of sorting information, in addition to experience from earlier visits, are signs and asking the personnel. Although the service at recycling centres is perceived positively by a majority of users, substantial improvements can still be made, and a number of such suggestions are given. PMID- 19674734 TI - Personal noise exposures of operators of agricultural tractors. AB - Approximately one million agricultural tractors are used in Turkey for crop production and about one-third of the population lives in rural areas. The objectives of this study were to determine sound pressure levels, A-weighted sound pressure levels, and the permissible exposure time for tractors without cabins, field-installed cabins, and original cabins at ear level of agricultural tractor operators for following machines: plows, cultivators, top soil cultivators, rotary tillers, tool combinations (harrow+roller), mechanical drills, pneumatic drills, chemical applicators, fertilizer applicators, drum mowers, balers, and forage harvesters. Variance analyses showed that type of operation, type of cabins, and operation x cabin interactions were statistically significant (P<0.01) both for sound pressure levels and equivalent (A-weighted) sound pressure levels. The use of original cabins had a greater effect in decreasing average sound pressures and resulted in more efficient noise insulation, especially at higher center frequencies compared to field-installed cabins whereas field-installed cabins proved to be more favorable compared to tractors without cabins. Sound pressure levels at 4000Hz center frequency was reduced 2-13dB and 4-18dB by using a field-installed cabin and an original cabin, respectively. The measured A-weighted equivalent sound pressure levels were compared to the threshold limit level, and was concluded that depending on the cabin types used, the operators could usually work from 4 to 6h a day without suffering from noise induced inconveniences while 2-3h is permissible for plowing and forage harvesting on tractors without cabins. Due to timeliness considerations in agricultural machine operations, a farmer would not be willing to interrupt the operation based on permissible exposure time set by the standards. Based on the findings of this study, particularly an original cabin is recommended to reduce machine-induced noise below the danger limit during agricultural machine operations. Personal protection devices should be used when tractors are operated without cabins, which could reduce A-weighted equivalent sound pressure levels by 10-45dB(A). PMID- 19674737 TI - Do perfectionists raise their standards after success? An experimental examination of the revaluation of standard setting in perfectionism. AB - Setting high standards is one of the core aspects of perfectionism. Shafran, Cooper, and Fairburn (2002) proposed that individuals with self-imposed perfectionism raise their standards after they successfully meet them. This study examines whether individuals with self-imposed perfectionism raise their goal after the feedback of success. Fifty-three college students, whose self-oriented perfectionism (SOP; Hewitt & Flett, 1991) was assessed, performed a task with a goal, and received feedback of success. Following the feedback, they were asked to choose (a) the same goal or (b) a more difficult goal for the next task. The logistic regression analysis revealed that the greater the SOP, the more likely the participants were to choose the difficult goal. The theoretical and clinical implications for the standard setting of perfectionism were discussed. PMID- 19674736 TI - Cancer progression is associated with increased expression of basement membrane proteins in three-dimensional in vitro models of human oral cancer. AB - BACKGROUND: Although basement membrane was traditionally considered an inert barrier that tumour cells had to cross before invasion into the surrounding stroma, recent studies suggest that basement membrane components are not only degraded during tumour progression, but also newly synthesised at the invasive front. OBJECTIVE: This study aimed at evaluating (1) the expression of basement membrane proteins in human oral carcinogenesis and (2) the role that epithelial mesenchymal interactions play on it, by using an in vitro oral cancer progression model. MATERIAL AND METHODS: In vitro three-dimensional (3D) organotypic cultures of normal, early neoplastic and neoplastic human oral mucosa were developed by growing primary normal human oral keratinocytes, dysplastic human oral keratinocytes (DOK cell line), and neoplastic human oral keratinocytes (PE/CA PJ15 cell line) on type I collagen biomatrices, with or without primary fibroblasts isolated from normal human oral mucosa. The cultured tissues were immunohistochemically assessed for the expression of the major basement membrane proteins laminin-332, type IV collagen, and fibronectin. RESULTS: Expression of laminin-332, type IV collagen, and fibronectin was gradually more pronounced in neoplastic models when compared to normal mucosa models, and, with the exception of laminin-332, it was further enhanced by presence of fibroblasts. Deposition of type IV collagen at the epithelium-biomatrix interface occurred only in presence of fibroblasts, as well as the extracellular matrix deposition of fibronectin. CONCLUSIONS: These findings, obtained in a 3D in vitro model that closely mirrors the in vivo human oral cancer progression, show an enhanced basement membrane protein expression during human oral cancer progression that is dependent on the epithelial-mesenchymal environment, respectively the existence of fibroblasts. PMID- 19674739 TI - Information learned from generic language becomes central to children's biological concepts: evidence from their open-ended explanations. AB - Generic sentences (e.g., "Snakes have holes in their teeth") convey that a property (e.g., having holes in one's teeth) is true of a category (e.g., snakes). We test the hypothesis that, in addition to this basic aspect of their meaning, generic sentences also imply that the information they express is more conceptually central than the information conveyed in similar non-generic sentences (e.g., "This snake has holes in his teeth"). To test this hypothesis, we elicited 4- and 5-year-old children's open-ended explanations for generic and non-generic versions of the same novel properties. Based on arguments in the categorization literature, we assumed that, relative to more peripheral properties, properties that are understood as conceptually central would be explained more often as causes and less often as effects of other features, behaviors, or processes. Two experiments confirmed the prediction that preschool age children construe novel information learned from generics as more conceptually central than the same information learned from non-generics. Additionally, Experiment 2 suggested that the conceptual status of novel properties learned from generic sentences becomes similar to that of familiar properties that are already at the category core. These findings illustrate the power of generic language to shape children's concepts. PMID- 19674738 TI - Memory for emotional faces in naturally occurring dysphoria and induced sadness. AB - The aim was to establish if the memory bias for sad faces, reported in clinically depressed patients (Gilboa-Schechtman, Erhard Weiss, & Jeczemien, 2002; Ridout, Astell, Reid, Glen, & O'Carroll, 2003) generalizes to sub-clinical depression (dysphoria) and experimentally induced sadness. Study 1: dysphoric (n=24) and non dysphoric (n=20) participants were presented with facial stimuli, asked to identify the emotion portrayed and then given a recognition memory test for these faces. At encoding, dysphoric participants (DP) exhibited impaired identification of sadness and neutral affect relative to the non-dysphoric group (ND). At memory testing, DP exhibited superior memory for sad faces relative to happy and neutral. They also exhibited enhanced memory for sad faces and impaired memory for happy relative to the ND. Study 2: non-depressed participants underwent a positive (n=24) or negative (n=24) mood induction (MI) and were assessed on the same tests as Study 1. At encoding, negative MI participants showed superior identification of sadness, relative to neutral affect and compared to the positive MI group. At memory testing, the negative MI group exhibited enhanced memory for the sad faces relative to happy or neutral and compared to the positive MI group. CONCLUSION: MCM bias for sad faces generalizes from clinical depression to these sub-clinical affective states. PMID- 19674740 TI - Real time acquisition and PC to PC wireless transmission of human carotid pulse waveform. AB - Carotid pulse is an established physiological parameter used to deduce cardiac information. Technological advances allow non-invasive measurement of carotid waveform, but involve complex electronics. Simple and relatively inexpensive method is devised here to acquire the carotid pulse. Measurements were made non invasively on human subjects in various postures using piezoelectric sensor. Transducer output is interfaced directly to a computer in real time and carotid wave is viewed successfully on virtual oscilloscope. Stored data is digitally filtered, visually analyzed and transferred wirelessly from one computer to another using windows hyper-terminal and XBee RF transceiver module. Automated analysis algorithm can be further developed. PMID- 19674741 TI - Robust and fast shell registration in PET and MR/CT brain images. AB - A robust and fast hybrid method using a shell volume that consists of high contrast voxels with their neighbors is proposed for registering PET and MR/CT brain images. Whereas conventional hybrid methods find the best matched pairs from several manually selected or automatically extracted local regions, our method automatically selects a shell volume in the PET image, and finds the best matched corresponding volume using normalized mutual information (NMI) in overlapping volumes while transforming the shell volume into an MR or CT image. A shell volume not only can reduce irrelevant corresponding voxels between two images during optimization of transformation parameters, but also brings a more robust registration with less computational cost. Experimental results on clinical data sets showed that our method successfully aligned all PET and MR/CT image pairs without losing any diagnostic information, while the conventional registration methods failed in some cases. PMID- 19674742 TI - Age-related changes in prefrontal cortex activity are associated with behavioural deficits in both temporal and spatial context memory retrieval in older adults. AB - Aging is associated with decrements in both spatial and temporal context retrieval. Functional neuroimaging studies of young adults suggest that there are differences in left versus right lateral prefrontal cortex (PFC) contributions to spatial versus temporal source (recency) retrieval, respectively. The goal of the current study was to determine if age-related decreases in temporal and spatial context retrieval are due to common or distinct changes in PFC function. To address this goal we conducted an event-related functional magnetic resonance imaging (fMRI) study in which young and older adults performed recognition, recency and spatial context retrieval tasks using face stimuli to identify event related PFC regions associated with these retrieval tasks in both age groups. Our behavioural results indicated that older adults did not differ on recognition performance, but did exhibit a deficit in both context retrieval tasks, compared to young adults. The fMRI results suggest that age-related deficits in both spatial and temporal context retrieval may be linked to functional changes in right dorsolateral and left medial anterior PFC (APFC) function. In addition, based on brain-behaviour correlations in older adults, our results imply that older adults attempt to compensate for these deficits by engaging left dorsolateral PFC during spatial context retrieval and right APFC during temporal context retrieval. PMID- 19674743 TI - Assessment of long-term subacute exposure to dimethoate by hair analysis of dialkyl phosphates DMP and DMTP in exposed rabbits: The effects of dose, dose duration and hair colour. AB - Hair analysis for dialkyl phosphates' (DAPs) residues could provide a measure of chronic exposure to organophosphate pesticides (OPs). The aim of this study was to determine whether these metabolites can be internally incorporated into the hair of rabbits exposed to dimethoate and also to investigate the influence of dose and dose duration of this OP, as well as the effect of hair colour on the concentrations of its DAPs in hair. Two-coloured rabbits were daily exposed to dimethoate (0, 12 or 24mgkg(-1) body weight) via their drinking water. Hair samples of both colours were obtained 4 and 6 months after the beginning of exposure from the back of all treated rabbits, and each hair colour sample was analyzed for dimethyl phosphate (DMP) and dimethyl thiophosphate (DMTP) by gas chromatography-mass spectrometry (GC-MS). Analysis revealed the incorporation of these metabolites into the rabbit hair in a dose-dependent manner. The mean concentrations found ranged from 0.18 to 0.77ngmg(-1) for DMP and from 0.43 to 1.53ngmg(-1) for DMTP. Mixed results for the significance of the relationship between dose duration and the levels of the two DAPs in hair are observed. Hair pigmentation does not appear to affect the concentration values of DMTP, whereas it seems to be a critical factor in the incorporation of DMP into hair. These data confirm the ability of hair testing to assess chronic OP exposure by the detection of DAPs. PMID- 19674744 TI - Response to a cardiac event in relation to cardiac knowledge and risk perception in a Lebanese sample: a cross sectional survey. AB - BACKGROUND: Heart disease is the leading cause of morbidity and mortality in many countries, including Lebanon. The continuing advances in the treatment of heart disease have improved patient outcomes significantly over the years. Nevertheless, prompt treatment remains essential to treatment success, which depends on the person's early recognition of cardiac symptoms and quick seeking of care. OBJECTIVE: The purpose of the study was to examine the relationship between knowledge of symptoms of heart disease, perceived cardiac risk and the potential response to a heart attack in a sample of Lebanese adults. DESIGN: A cross sectional descriptive survey design was used. A convenience sample of 399 adult hospital visitors who did not have heart disease were interviewed in two major referral hospitals in Beirut. MEASURES: The Illness Perception Questionnaire and the Behavioral Risk Factor Surveillance System were used; in addition an open-ended question was asked to elicit what the visitor would do in case he/she experienced cardiac symptoms. RESULTS: While most participants (94%) correctly recognized major symptoms of heart disease (like chest pain and dyspnea), they also reported irrelevant symptoms to be associated with heart disease (like irritability by 68% and muscle cramps by 52%). Only 21% of participants reported that they would seek emergency care as their first response if they were experiencing a heart attack. Knowledge of symptoms and perceived cardiac risk were not associated with the response to a heart attack. Being single was the only significant predictor of choosing the appropriate response when experiencing a cardiac event. CONCLUSION: The findings suggest lack of adequate knowledge of cardiac symptoms and a high potential for inappropriate responses when symptoms occur. Public health heart health campaigns are needed to raise awareness about heart disease in Lebanon. PMID- 19674727 TI - SCORE Study Report 7: incidence of intravitreal silicone oil droplets associated with staked-on vs luer cone syringe design. AB - PURPOSE: To evaluate the incidence of intravitreal silicone oil (SO) droplets associated with intravitreal injections using a staked-on vs luer cone syringe design in the SCORE (Standard Care vs COrticosteroid in REtinal Vein Occlusion) Study. DESIGN: Prospective, randomized, phase III clinical trial. METHODS: The incidence of intravitreal SO was compared among participants exposed to the staked-on syringe design, the luer cone syringe design, or both of the syringe designs in the SCORE Study, which evaluated intravitreal triamcinolone acetonide injection(s) for vision loss secondary to macular edema associated with central or branch retinal vein occlusion. Injections were given at baseline and 4-month intervals, based on treatment assignment and study-defined retreatment criteria. Because intravitreal SO was observed following injections in some participants, investigators were instructed, on September 22, 2006, to look for intravitreal SO at all study visits. On November 1, 2007, the luer cone syringe design replaced the staked-on syringe design. RESULTS: A total of 464 participants received a total of 1,205 injections between November 4, 2004 and February 28, 2009. Intravitreal SO was noted in 141 of 319 participants (44%) exposed only to staked on syringes, 11 of 87 (13%) exposed to both syringe designs, and 0 of 58 exposed only to luer cone syringes (P < .0001). Among participants with first injections after September 22, 2006, intravitreal SO was noted in 65 of 114 (57%) injected only with staked-on syringes compared with 0 of 58 injected only with luer cone syringes. Differential follow-up is unlikely to explain these results. CONCLUSION: In the SCORE Study, luer cone syringe design is associated with a lower frequency of intravitreal SO droplet occurrence compared with the staked-on syringe design, likely attributable to increased residual space in the needle hub with the luer cone design. PMID- 19674745 TI - Circulating level of microparticles and their correlation with arterial elasticity and endothelium-dependent dilation in patients with type 2 diabetes mellitus. AB - OBJECTIVE: This study was designed to measure the circulating level of microparticles (MP) in patients with type 2 diabetes mellitus versus healthy volunteers and to evaluate their correlation with arterial elasticity and endothelium-dependent dilation. METHODS: Flow cytometry was used to measure the circulating levels of MP, including annexinV+MP, platelet-derived microparticles (PMP), leukocyte-derived microparticles (LMP) and endothelial microparticles (EMP), in 63 patients with type 2 diabetes mellitus and 29 healthy volunteers. Brachial ankle pulse wave velocity (baPWV) and endothelium-dependent flow mediated dilation (FMD) of the brachial artery were also measured. RESULTS: The levels of annexinV+MP, PMP, LMP, CD31+/CD42-EMP and CD51+EMP increased significantly in diabetic patients compared with healthy controls (P<0.001). Correlation analysis showed that HbA(1c) was positively correlated to CD31+/CD42 EMP (r(s)=0.337, P=0.008) and CD51+EMP (r(s)=0.266, P=0.038). FMD in diabetic patients was significantly lower than that in healthy individuals (P=0.007). FMD was negatively correlated to CD31+/CD42-EMP (r(s)=-0.441, P=0.008) and CD51+EMP (r(s)=-0.405, P=0.016). baPWV level in diabetic patients was significantly higher than that in healthy individuals (P<0.001). baPWV was positively correlated to CD31+/CD42-EMP (r(s)=0.497, P<0.001) and CD51+EMP (r(s)=0.428, P=0.001). Multiple regression analysis indicated that EMP was an independent risk factor of FMD and baPWV. CONCLUSIONS: The circulating level of microparticles increases in patients with type 2 diabetes. The level of endothelial microparticles is closely associated with vascular dysfunction. PMID- 19674746 TI - High plasma levels of matrix metalloproteinase-8 in patients with unstable angina. AB - Matrix metalloproteinases (MMPs) play a role in collagen breakdown, leading to plaque instability. High levels of MMPs mRNA and proteins, especially MMP-1, MMP 2, MMP-8, MMP-9, and MMP-13, were shown in human atherosclerotic plaques. However, among various MMPs, only MMP-1, MMP-8 and MMP-13, so-called interstitial collagenases, can initiate collagen breakdown. To elucidate whether MMP-1, MMP-8 and MMP-13 levels in blood were high in patients with unstable angina (UAP), we measured serum MMP-1 and plasma MMP-8 and MMP-13 levels in 45 patients with UAP, 175 with stable coronary artery disease (CAD), and 45 controls. Plasma C-reactive protein levels tended to be higher in patients with UAP than in those with stable CAD and controls (median 0.94 vs. 0.69 and 0.51mg/l). Regarding blood levels of MMPs, MMP-13 levels were above the lower detection limit in only one patient with UAP (2%), one with stable CAD (1%), and none in controls. MMP-1 levels did not differ among patients with UAP, stable CAD, and controls (median 4.8, 5.3, and 5.4ng/ml). Notably, MMP-8 levels were higher in patients with stable CAD than in controls (median 3.5ng/ml vs. 2.8ng/ml, P<0.005), however, MMP-8 levels in patients with UAP were much higher than those in stable CAD (3.9ng/ml vs. 3.5ng/ml, P<0.05). In multivariate analysis, MMP-8 level was an independent factor for UAP. Thus, plasma MMP-8 levels were found to be high in patients with UAP, suggesting that MMP-8 levels in UAP may reflect coronary plaque instability and that MMP-8 is a promising biomarker for UAP. PMID- 19674748 TI - An application of principal component analysis for lower body kinematics between loaded and unloaded walking. AB - Load carriage is a very common daily activity at home and in the workplace. Generally, the load is in the form of an external load carried by an individual, it could also be the excessive body mass carried by an overweight individual. To quantify the effects of carrying extra weight, whether in the form of an external load or excess body mass, motion capture data were generated for a diverse subject set. This consisted of twenty-three subjects generating one hundred fifteen trials for each loading condition. This study applied principal component analysis (PCA) to motion capture data in order to analyze the lower body gait patterns for four loading conditions: normal weight unloaded, normal weight loaded, overweight unloaded and overweight loaded. PCA has been shown to be a powerful tool for analyzing complex gait data. In this analysis, it is shown that in order to quantify the effects of external loads and/or for both normal weight and overweight subjects, the first principal component (PC1) is needed. For the work in this paper, PCs were generated from lower body joint angle data. The PC1 of the hip angle and PC1 of the ankle angle are shown to be an indicator of external load and BMI effects on temporal gait data. PMID- 19674749 TI - Tolerance of the skull to blunt ballistic temporo-parietal impact. AB - Less-lethal ballistic projectiles are used by police personnel to temporarily incapacitate suspects. While the frequency of these impacts to the head is low, they account for more serious injuries than impacts to any other body region. As a result, there is an urgent need to assess the tolerance of the head to such impacts. The focus of this study was to investigate the tolerance of the temporo parietal skull to blunt ballistic impact and establish injury criteria for risk assessment. Seven unembalmed isolated cadaver heads were subjected to fourteen impacts. Specimens were instrumented with a nine-accelerometer array as well as strain gages surrounding the impact site. Impacts were performed with a 38 mm instrumented projectile at velocities ranging from 18 to 37 m/s. CT images and autopsies were performed to document resulting fractures. Peak fracture force for the seven resulting fractures was 5633+/-2095 N. Peak deformation for fracture producing impacts was 7.8+/-3.2 mm. The blunt criterion (BC), peak force and principal strain were determined to be the best predictors of depressed comminuted fractures. Temporo-parietal tolerance levels were consistent with previous studies. An initial force tolerance level of 2346 N is established for the temporo-parietal region for blunt ballistic impact with a 38 mm diameter impactor. PMID- 19674747 TI - Non-nucleoside reverse transcriptase inhibitor efavirenz increases monolayer permeability of human coronary artery endothelial cells. AB - Highly active antiretroviral therapy (HAART) is often associated with endothelial dysfunction and cardiovascular complications. In this study, we determined whether HIV non-nucleoside reverse transcriptase inhibitor efavirenz (EFV) could increase endothelial permeability. Human coronary artery endothelial cells (HCAECs) were treated with EFV (1, 5 and 10 microg/ml) and endothelial permeability was determined by a transwell system with a fluorescence-labeled dextran tracer. HCAECs treated with EFV showed a significant increase of endothelial permeability in a concentration-dependent manner. With real time PCR analysis, EFV significantly reduced the mRNA levels of tight junction proteins claudin-1, occludin, zonula occluden-1 and junctional adhesion molecule-1 compared with controls (P<0.05). Protein levels of these tight junction molecules were also reduced substantially in the EFV-treated cells by western blot and flow cytometry analyses. In addition, EFV also increased superoxide anion production with dihydroethidium and cellular glutathione assays, while it decreased mitochondrial membrane potential with JC-staining. Antioxidants (ginkgolide B and MnTBAP) effectively blocked EFV-induced endothelial permeability and mitochondrial dysfunction. Furthermore, EFV increased the phosphorylation of MAPK JNK and IkappaBalpha, thereby increasing NFkappaB translocation to the nucleus. Chemical JNK inhibitor and dominant negative mutant JNK and IkappaBalpha adenoviruses effectively blocked the effects of EFV on HCAECs. Thus, EFV increases endothelial permeability which may be due to the decrease of tight junction proteins and the increase of superoxide anion. JNK and NFkappaB activation may be directly involved in the signal transduction pathway of EFV action in HCAECs. PMID- 19674750 TI - Modeling aqueous humor collection from the human eye. AB - Glaucoma is a common cause of blindness. Studies of this disease can involve collection of aqueous humor (AH) fluid from eyes of patients undergoing surgery, which involves aspirating a small fluid volume from the anterior region of the eye through a fine-bore needle. Unfortunately, the composition of the AH is spatially non-uniform in the eye, and thus the composition of the aspirated fluid is uncertain. Our goal was to numerically simulate the AH aspiration process to determine where the aspirated fluid was being collected from and thus gain insight into the composition of the collected fluid. A 3D computational model of the anterior region of the human eye was created and the Navier-Stokes equations were numerically solved during the aspiration process for a set of typical (baseline) conditions: 40 microl aspirated volume and needle placement in the central anterior chamber. We also ran variations of this baseline simulation. The main finding was that the aspirated fluid comes from a very localized region around the needle tip, so that for typical conditions, almost no aspirated fluid is withdrawn from the angle region of the anterior chamber. This is important because the AH in this angle region is protein-rich and directly interacts with the tissues that control fluid drainage from the eye. Recommendations for standardizing aspiration conditions are given. PMID- 19674751 TI - Ultrasound-assisted dispersive liquid-liquid microextraction combined with high performance liquid chromatography-fluorescence detection for sensitive determination of biogenic amines in rice wine samples. AB - Ultrasound-assisted dispersive liquid-liquid microextraction coupled with high performance liquid chromatography-fluorescence detection was used for the extraction and determination of three biogenic amines including octopamine, tyramine and phenethylamine in rice wine samples. Fluorescence probe 2,6-dimethyl 4-quinolinecarboxylic acid N-hydroxysuccinimide ester was applied for derivatization of biogenic amines. Acetonitrile and 1-octanol were used as disperser solvent and extraction solvent, respectively. Extraction conditions including the type of extraction solvent, the volume of extraction solvent, ultrasonication time and centrifuging time were optimized. After extraction and centrifuging, analyte was injected rapidly into high-performance liquid chromatography and then detected with fluorescence. The calibration graph of the proposed method was linear in the range of 5-500 microg mL(-1) (octopamine and tyramine) and 0.025-2.5 microg mL(-1) (phenethylamine). The relative standard deviations were 2.4-3.2% (n=6) and the limits of detection were in the range of 0.02-5 ng mL(-1). The method was applied to analyze the rice wine samples and spiked recoveries in the range of 95.42-104.56% were obtained. The results showed that ultrasound-assisted dispersive liquid-liquid microextraction was a very simple, rapid, sensitive and efficient analytical method for the determination of trace amount of biogenic amines. PMID- 19674753 TI - One-step in-syringe ionic liquid-based dispersive liquid-liquid microextraction. AB - Dispersive liquid-liquid microextraction (DLLME) has been proved to be a powerful tool for the rapid sample treatment of liquid samples providing at the same time high enrichment factors and extraction recoveries. A new, simple and easy to handle one step in-syringe set-up for DLLME is presented and critically discussed in this paper. The novel approach avoids the centrifugation step, typically off line and time consuming, opening-up a new horizon on DLLME automation. The suitability of the proposal is evaluated by means of the determination of non steroidal anti-inflammatory drugs in urine by liquid chromatography/ultraviolet detection. In the presented approach an ionic liquid is used as extractant. The target drugs can be determined in urine within the concentration range 0.02-10 microg mL(-1), allowing their determination at therapeutic and toxic levels. Limits of detection were in the range from 8.3 ng mL(-1) (indomethacin) to 32 ng mL(-1) (ketoprofen). The repeatability of the proposed method expressed as RSD (n=5) varied between 2.5% (for ketoprofen) and 8.6% (for indomethacin). PMID- 19674752 TI - Ultrasound-assisted emulsification microextraction method based on applying low density organic solvents followed by gas chromatography analysis for the determination of polycyclic aromatic hydrocarbons in water samples. AB - In this study, a fast, simple and efficient ultrasound-assisted emulsification microextraction (USAEME) method was successfully developed based on applying low density organic solvents. Fourteen microliters of toluene was injected slowly into a 12 mL home-designed centrifuge glass vial containing an aqueous sample that was located inside the ultrasonic water bath. The formed emulsion was centrifuged and 2 microL of separated toluene (about 4 microL) was injected into a gas chromatographic system equipped with a flame ionization detector (GC-FID) for analysis. Some polycyclic aromatic hydrocarbons (PAHs) were selected as model compounds for developing the method and evaluating its performance and to compare the efficiency of the proposed method with previously reported techniques. Several factors influencing the emulsification, extraction and collection efficiency such as the nature and volume of organic solvent, emulsification extraction temperature, ionic strength and equilibrium and centrifugation times were investigated and optimized. Under the optimum conditions, preconcentration factors (PFs) in a range of 1776-2714 were obtained. The performance of the proposed method was studied in terms of linear dynamic range (LDRs from 0.05 to 100 microg L(-1)), linearity (R(2) > or = 0.994), precision (repeatability: RSD% < or = 7.9, reproducibility: RSD% < or = 14.6) and extraction percents (59.2 90.5%). Limits of detection (LODs) in the range of 0.02-0.05 microg L(-1) were obtained for different PAHs. The applicability of the proposed method was evaluated by the extraction and determination of PAHs from several natural water samples. PMID- 19674754 TI - Synthesis and characterization of the water-soluble silica-coated ZnS:Mn nanoparticles as fluorescent sensor for Cu(2+) ions. AB - Silica-coated ZnS:Mn nanoparticles were synthesized by coating hydrophobic ZnS:Mn nanoparticles with silica shell through microemulsion. The core-shell structural nanoparticles were confirmed by X-ray diffraction (XRD) patterns, high-resolution transmission electron microscope (HRTEM) images and energy dispersive spectroscopy (EDS) measurements. Results show that each core-shell nanoparticle contains single ZnS:Mn nanoparticle within monodisperse silica nanospheres (40nm). Photoluminescence (PL) spectroscopy and UV-vis spectrum were used to investigate the optical properties of the nanoparticles. Compared to uncoated ZnS:Mn nanoparticles, the silica-coated ZnS:Mn nanoparticles have the improved PL intensity as well as good photostability. The obtained silica-coated ZnS:Mn nanoparticles are water-soluble and have fluorescence sensitivity to Cu(2+) ions. Quenching of fluorescence intensity of the silica-coated nanoparticles allows the detection of Cu(2+) concentrations as low as 7.3x10(-9)molL(-1), thus affording a very sensitive detection system for this chemical species. The possible quenching mechanism is discussed. PMID- 19674755 TI - Carboxylated core-shell particles: I. A system showing hindered swelling behavior. AB - We study the swelling behavior of carboxylated core-shell particles. It is well known that these particles swell with increasing pH due to the electrostatic repulsion between carboxylate groups. Our results reveal that the swelling behavior is affected by the preparation method. We find that the swelling is promoted in those particles which were initially in a highly swollen state (pH>or=10). However, the swelling is hindered for those particles which were not previously in this trigger pH. In the hindered systems, a compact conformation of the polymer shell is induced by hydrophobic attractions between the non-charged segments which compete against the swelling driving force. In addition, an interesting hysteresis behavior emerges when promoted systems are subjected to a heating-cooling cycle; a new stable system appears with a less extended polymer shell conformation. Furthermore, salt-induced swelling experiments corroborate not only polymer restructuring but also assembly among carboxylate groups which affects their ionization grade. PMID- 19674756 TI - Fairness considerations: increasing understanding of intentionality during adolescence. AB - The development of fairness considerations in decision making is not well understood. Here we tested the hypothesis that increased understanding of intentionality during adolescence underlies increased fairness considerations in social decision making. We conducted three experiments using an adapted version of the Ultimatum Game with participants during four stages of adolescence: 9, 12, 15, and 18 years of age. Participants made or evaluated monetary offers, and we manipulated the intentionality context of offers. Results show that strategic thinking is already present at 9 years of age. There was no age difference in fairness of offers when the responder could not reject an offer (Experiment 1), but when they could reject an offer there was an age-related increase in taking into account the perspective and intentionality of other players when making offers (Experiment 2) and evaluating offers (Experiment 3). Taken together, the results demonstrate a linear developmental transition in fairness considerations that may have important implications for our understanding of social interactions during adolescence. PMID- 19674757 TI - Gender differences in the histopathology of MS? AB - Multiple sclerosis (MS) lesions are histopathologically characterized by inflammation, demyelination/remyelination, axonal damage and gliosis. Animal experimental and in vitro studies suggest that sex hormones influence the immune system and contribute to the increased likelihood in women of developing MS. However, a variety of studies have also shown that remyelination is more marked in female rodents or that female sex hormones are beneficial for myelin repair. To determine whether gender influences the histopathology of MS lesions, we compared the extent of inflammation, axonal damage and remyelination in MS lesions of female and male MS patients. We observed no differences in the composition of inflammatory infiltrates, axonal damage or cortical pathology. Similar numbers of oligodendroglial progenitor cells and mature oligodendrocytes were present in MS lesions. Remyelination is slightly, but not significantly, more extensive in women than men in early MS lesions. The absence of significant differences in lesion pathology between female and male MS patients might be explained by a lack of a gender influence, but also might be due to the limited number of tissue samples available for histopathological analysis. PMID- 19674758 TI - Clustering of multiple sclerosis, age of onset and gender in Sardinia. AB - In diseases with unpredictably long latent period, such as multiple sclerosis (MS), cluster studies, and the analysis of disease distribution by gender and age of onset, can provide clues to disease etiopathogenesis. Sardinia, insular Italy, is among the regions at highest risk for MS worldwide. Multiple epidemiological evidences favour the role of exogenous factors in modulating MS incidence in Sardinians. Its steady increase over time is the main determinant for the observed increased prevalence of MS. Incidence is especially increasing for MS with RR initial course, in individuals with older age and in the female population. More women that will develop MS are believed to being born now. Early childhood is likely the age of susceptibility to MS, and especially in women, in most recent cases, and in individuals developing MS at earlier age and with RR initial clinical course. These evidences suggest that specific exogenous (gender related? perinatal?) factors have recently been exerting as determinants of the MS increase in Sardinians and the change of its phenotype. Such factors are unlikely agents with fixed latency incubation period, but they could rather be the result of recent lifestyle changes and/or of their interplay with a highly susceptible genetic background. PMID- 19674759 TI - [Implantation of aortic valvular prosthesis via transfemoral catheter. Evaluation of candidates undergoing the procedure]. AB - BACKGROUND AND OBJECTIVE: In recent years, techniques for implantation of aortic prosthesis via catheter have been developed as a therapeutic alternative in patients with severe aortic stenosis rejected for surgery. The correct selection of candidates is one of the more complex aspects of this treatment. We analyzed the acceptance rate in our environment for transcatheter aortic valve implantation in patients referred to our hospital for evaluation as possible candidates, describing the exclusion reasons. PATIENTS AND METHOD: 30 patients with severe aortic stenosis and rejected for surgical aortic valve replacement were referred to our hospital to evaluate transcatheter aortic valve implantation. The patients first underwent clinical evaluation and were studied with echocardiography, angiography and computed tomography. RESULTS: Of the 30 patients, 18 were rejected for the procedure (60%): 4 patients with non-severe aortic stenosis, 2 asymptomatic patients, 2 patients who finally underwent surgery because of a low-surgical-risk, 5 patients with contraindications for the procedure, 2 patients who finally did not want to undergo the procedure and 3 patients were further rejected because the vascular access was inappropriate. Of the remaining 12 patients initially accepted, 3 died before the procedure was performed. Finally, only 9 patients (30%) underwent transcatheter aortic valve implantation. CONCLUSIONS: Of the patients referred for transcatheter aortic valve implantation, only 40% were accepted. The mortality rate during the evaluation process of this procedure is high, showing that these patients are terminally ill. PMID- 19674761 TI - [Impact of malnutrition on clinical outcome in hospitalized elderly patients]. PMID- 19674760 TI - [Differential aspects in children and adult patients with medulloblastoma]. AB - BACKGROUND AND OBJECTIVE: Medulloblastoma is the more frequent malignant cerebral tumor in childhood. PATIENTS AND METHODS: This is an observational study of a retrospective cohort in which there were included all the patients diagnosed of medulloblastoma in the last 19 years (1989-2007) in Hospital de Cruces of Baracaldo, Vizcaya, Spain. RESULTS: There were included 37 patients, 20 men and 17 women, with ages between 1 and 48 years (average age 13.7 years with standard deviation 11.4). Tumor site and mortality according to initial dissemination were variables of statistic significance. Metastases were detected at diagnosis in two patients. The surgical resection was total in 75% of the patients. A relapse was diagnosed in the follow-up in 59.5% of the patients, with a positive spinal fluid in 27%. Sequelae were detected in 100% of the survivors, mainly with cerebellar and ocular alterations. One patient developed a meningioma and a maxillary sarcoma at the long term follow-up. It is important to emphasize the aggressiveness of medulloblastoma in both children and adults, with a global mortality of 56.8% and 48,6% at 5 years. CONCLUSIONS: It is considered necessary a multidisciplinary treatment and a long term monitoring of the patients and the sequelae of the survivors, including the possibility of second tumours. PMID- 19674762 TI - PAF-antagonistic bicyclo[3.2.1]octanoid neolignans from leaves of Ocotea macrophylla Kunth. (Lauraceae). AB - Di-nor-benzofuran neolignan aldehydes, Delta(7)-3,4-methylenedioxy-3'-methoxy 8',9'-dinor-4',7-epoxy-8,3'-neolignan-7'-aldehyde (ocophyllal A) 1, Delta(7) 3,4,5,3'-tetramethoxy-8',9'-dinor-4',7-epoxy-8,3'-neolignan-7'-aldehyde (ocophyllal B) 2, and macrophyllin-type bicyclo[3.2.1]octanoid neolignans (7R, 8R, 3'S, 4'S, 5'R)-Delta(8)'-4'-hydroxy-5'-methoxy-3,4-methylenedioxy-2',3',4',5' tetrahydro-2'-oxo-7.3',8.5'-neolignan (ocophyllol A) 3, (7R, 8R, 3'S, 4'S, 5'R) Delta(8)'-4'-hydroxy-3,4,5'-trimethoxy-2',3',4',5'-tetrahydro-2'-oxo-7.3',8.5' neolignan (ocophyllol B) 4, (7R, 8R, 3'S, 4'S, 5'R)-Delta(8)'-4'-hydroxy-3,4,5,5' tetramethoxy-2',3',4',5'-tetrahydro-2'-oxo-7.3',8.5'-neolignan (ocophyllol C) 5, as well as 2'-epi-guianin 6 and (+)-licarin B 7, were isolated and characterized from leaves of Ocotea macrophylla (Lauraceae). The structures and configuration of these compounds were determined by extensive spectroscopic analyses. Inhibition of platelet activating factor (PAF)-induced aggregation of rabbit platelets were tested with neolignans 1-7. Although compound 6 was the most potent PAF-antagonist, compounds 3-5 showed some activity. PMID- 19674763 TI - Prevalence of and risk factors for fatty liver disease in a professional population of Wuhan, China. AB - OBJECTIVES: To explore the prevalence of fatty liver disease (FLD) and its risk factors in a cold steel-rolling mill in Wuhan, China. STUDY DESIGN: A cross sectional survey. METHODS: In total, 1174 subjects were investigated for FLD (596 exposed workers and 578 controls). FLD was determined by ultrasonography. Height, weight, blood pressure, fasting plasma glucose, serum aspartate aminotransferase (AST), serum triglyceride (TG) and serum hepatitis B surface antigen (HBsAg) were determined by routine laboratory methods. RESULTS: The prevalences of major diseases such as FLD (41.6%), hypertension (21.1%) and high serum TG (42.6%) were higher in the exposed workers than in the controls. The prevalence of FLD was highest among grinding roller workers (50.6%); this was almost four times as high as in the control group (14.7%). Logistic regression analysis indicated that all six types of work (zincing, shearing, tinning, operating grinding rollers, electrics and fitting), as well as high serum TG, high serum AST, hyperglycaemia, hypertension, higher body mass index (BMI) and drinking alcohol were risk factors for FLD. CONCLUSION: A high prevalence of FLD was found among the exposed workers, and some risk factors, especially different types of work, were related to the prevalence of FLD. High serum TG, high serum AST, hypertension, hyperglycaemia, higher BMI and drinking alcohol were important risk factors for FLD. PMID- 19674764 TI - Occurrence and removal of pharmaceuticals, caffeine and DEET in wastewater treatment plants of Beijing, China. AB - The occurrence and removal of 13 pharmaceuticals and 2 consumer products, including antibiotic, antilipidemic, anti-inflammatory, anti-hypertensive, anticonvulsant, stimulant, insect repellent and antipsychotic, were investigated in four wastewater treatment plants (WWTPs) of Beijing, China. The compounds were extracted from wastewater samples by solid-phase extraction (SPE) and analyzed by ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS). Most of the target compounds were detected, with the concentrations of 4.4 ng L(-1)-6.6 microg L(-1) and 2.2-320 ng L(-1) in the influents and secondary effluents, respectively. These concentrations were consistent with their consumptions in China, and much lower than those reported in the USA and Europe. Most compounds were hardly removed in the primary treatment, while their removal rates ranging from -12% to 100% were achieved during the secondary treatment. In the tertiary treatment, different processes showed discrepant performances. The target compounds could not be eliminated by sand filtration, but the ozonation and microfiltration/reverse osmosis (MF/RO) processes employed in two WWTPs were very effective to remove them, showing their main contributions to the removal of such micro-pollutants in wastewater treatment. PMID- 19674765 TI - Treated water quality assurance and description of distribution networks by multivariate chemometrics. AB - Throughout the year 2007, 89 treated water samples from three water treatment plants (WTPs) of the Athens Water Supply and Sewerage Company (EYDAP S.A.) and 180 samples from network tanks (NWTs) were analyzed for electrical conductivity (EC), alkalinity (TA), pH, aluminium (Al), total hardness (TH), chloride (Cl(-)), residual chlorine (free Cl), calcium (Ca(2+)) and magnesium (Mg(2+)). The results regarding the WTPs were subjected to a principal component analysis (PCA) with 75% of the total variance being explained. A stepwise linear discriminant analysis (LDA) model constructed from the 89 treated water samples was used to predict class membership of the samples from the NWTs with a view to estimating the propagation of a possible water quality deterioration originating from the WTPs. The model utilized Cl(-), Al and EC and yielded a 96% correct classification of the training dataset, whereas the cross-validation yielded a 94% correct classification. Network tank samples were 95% correctly classified with regard to their theoretically expected origin. The stepwise discriminant analysis based on separate covariance matrices of the canonical discriminant functions yielded a 98% correct classification of both the training dataset and the network tank samples. The classification and regression tree (C&RT) algorithm showed that the main parameters used in the discrimination of the WTP samples were EC and Al. The post-hoc classification of the training dataset was 99%, whereas 88% of NWT samples were correctly classified. PMID- 19674766 TI - Synthesis and NMR properties of derivatives of 5,6-dihydroborauracil and 5,6 dihydroborathymine. AB - Novel boron compounds, a series of 4-hydroxy-5,6-dihydroborauracil and 4-hydroxy 5,6-dihydroborathymine derivatives containing various substituents at 3-, 5- and 6-positions, is presented. The spectroscopic properties, along with analyses of NMR-controlled boron compound-alcohol and boron compound-amine interactions, proves the existence of sp(3)-hybridized, stable B,B-bis-methoxy-5,6 dihydroborauracils and pyridine-/n-butylamine-5,6-dihydroborauracils ate complexes in solution. PMID- 19674767 TI - Inactivation of Ascaris eggs in soil by microwave treatment compared to UV and ozone treatment. AB - This study reports on the effect of microwave radiation for inactivation of Ascaris lumbricoides eggs in 25 g of soil compared to ultraviolet irradiation and ozone expose. Microwave radiation at 700 W with 14% water content (w/w) achieved approximately 2.5 log inactivation of eggs in soil within 60s. On the other hand, UV irradiation at 3 mW cm(-2) with and without shaking soil for 3600 s achieved approximately 0.32 and 0.01 log inactivation of eggs, respectively. In ozone treatment, 0.13 log inactivation of eggs was achieved with 5.8+/-0.7 mg L(-1) of dissolved ozone dose for 30 min in a continuous diffusion reactor. In addition, the inactivation of eggs by three disinfection techniques was conducted in water in order to compare the inactivation efficiency of eggs in soil. The inactivation efficiency of microwave radiation was found to be no significant difference between in soil and water. However, the inactivation efficiency of UV irradiation was significantly increased in water while in ozone expose there was no significant difference between in soil and water. Microwave treatment thus proved to be the most efficient method in controlling A. lumbricoides eggs in soil. PMID- 19674768 TI - Joint toxicity of fluoranthene and pentachlorobenzene to Hyalella azteca and Chironomus dilutus. AB - Nonpolar organic chemicals such as polycyclic aromatic hydrocarbons and chlorobenzenes are expected to act additively when exposed as a mixture. The present study examined the toxicity of fluoranthene (FLU) and pentachlorobenzene (PCBz) individually and in a binary mixture using the whole-body residue as the dose metric. Body residues were based on the toxic equivalent body residue, which included the parent compound plus the organically extractable metabolites for FLU and the parent compound only for PCBz. Using a toxic unit (TU) approach, the binary mixtures of FLU and PCBz following 4- and 10-d water-only exposures acted additively. The lethal residue (LR50) values for mixtures of the compounds for Hyalella azteca were 1.26 (1.19-1.33) TU and 1.27 (1.20-1.34) TU for 4- and 10-d exposures, respectively. For Chironomus dilutus, the 4-d and 10-d values were 0.93 (0.90-0.97) TU and 1.01 (0.96-1.06) TU. Additionally, the total molar sum of PCBz and FLU whole-body residues in a mixture were compared to residues from single compound exposures. For both species tested, the LR50 values based on the total molar sum fell within the range of those determined from the single compound tests; providing additional support for molar additivity for nonpolar narcotic compounds. Assuming that residue-effects data among narcotic compounds (e.g., LR50) are similar, applying the molar sum methodology to narcotic compounds in tissues determined from routine biomonitoring programs and risk specific sampling may be a valuable tool to assess potential effects to biota in the field. PMID- 19674769 TI - Atrazine and terbuthylazine mineralization by an Arthrobacter sp. isolated from a sugarcane-cultivated soil in Kenya. AB - A tropical soil from a Kenyan sugarcane-cultivated field showed a very high capability to mineralize (14)C-ring-labeled atrazine. In laboratory experiments this soil mineralized about 90% of the applied atrazine within 98 d. The atrazine degrading microbial community was enriched in liquid cultures containing atrazine as the sole N source and 100 mgL(-1) glucose as additional C source. From the enrichment culture a bacterial strain was isolated and identified by comparative sequence analysis of the 16S-rDNA as member of the genus Arthrobacter. The enriched mixed culture as well as the isolated strain, designated as Arthrobacter sp. strain GZK-1, could grow on atrazine and terbuthylazine as sole N-sources; Arthrobacter sp. GZK-1 mineralized (14)C-ring-labeled atrazine up to 88% to (14)CO(2) and (14)C-ring-labeled terbuthylazine up to 65% to (14)CO(2) in a liquid culture within 14 d. The enriched microbial consortium as well as the isolated strain could be a potential solution for the remediation of s-triazine polluted agricultural soils. PMID- 19674770 TI - Body size and its consequences: allometry and the lower limb length of Liang Bua 1 (Homo floresiensis). AB - Bivariate femoral length allometry in recent humans, Pan, and Gorilla is investigated with special reference to the diminutive Liang Bua (LB) 1 specimen (the holotype of Homo floresiensis) and six early Pleistocene femora referred to the genus Homo. Relative to predicted body mass, Pan and Gorilla femora show strong negative length allometry while recent human femora evince isometry to positive allometry, depending on sample composition and line-fitting technique employed. The allometric trajectories of Pan and Homo show convergence near the small body size range of LB 1, such that LB 1 manifests a low percentage deviation (d(yx) of Smith [1980]) from the Pan allometric trajectory and falls well within the 95% confidence limits around the Pan individuals (but also outside the 95% confidence limits for recent Homo). In contrast, the six early Pleistocene Homo femora, belonging to larger individuals, show much greater d(yx) values from both Pan and Gorilla and fall well above the 95% confidence limits for these taxa. All but one of these Pleistocene Homo specimens falls within the 95% confidence limits of the recent human sample. Similar results are obtained when femoral length is regressed on femoral head diameter in unlogged bivariate space. Regardless of the ultimate taxonomic status of LB 1, these findings are consistent with a prediction made by us (Franciscus and Holliday, 1992) that hominins in the small body size range of A.L. 288-1 ("Lucy"), including members of the genus Homo, will tend to possess short, ape-like lower limbs as a function of body size scaling. PMID- 19674771 TI - Effects of pre-ozonation on the removal of THM precursors by coagulation. AB - Pre-ozonation in combination with enhanced coagulation was used to remove NOM from lake water as to control the formation of disinfection by-products, DBPs. The effect of the hydrophobicity/hydrophilicity nature of NOM on the performance of the combined pre-ozonation and coagulation process was studied. The hydrophilicity/hydrophobicity property of NOM was characterized in terms of mass distribution of the phydrophilic and the hydrophobilic fractions of NOM. The optimal condition for the combined pre-ozonation-coagulation was established: pH=ca. 9.0 and ozone dose=0.45 mg-O(3)/mg-DOC. Under the optimal condition, it was able to achieve approximately 60% of THMFP removal. In terms of THMPF, results also indicated that the distribution between the hydrophilic and the hydrophobic fractions of NOM was 57.3 and 98.7 microg-THMFP/mg-DOC, respectively. Ozonation alters the structures and characteristics of NOM thereby affecting the coagulation effectiveness. Pre-ozonation was effective in removing the hydrophobic NOM, with a decrease of THMFP by approximately 20% versus approximately 10% for the hydrophilic fraction. The dosage of coagulant also governed DOC removal. The removal of hydrophobic and hydrophilic NOM were in the range of 27-41 and 2.5-22.7%, respectively at alum dosage of 0.41-1.65 (in Al/DOC) and 0.41-1.65 (in Al/DOC) and ozone dose of 0.58-2.93, mg/mg respectively. The adsorption characteristics of the hydrophilic and the hydrophobic fractions of NOM on aluminum hydroxide (from coagulant alum) were studied. Results indicated that the modified Langmuir isotherm of competitive adsorption was able to describe the adsorption of NOM onto hydrous aluminum hydroxide formed during alum coagulation of the lake waters. PMID- 19674772 TI - Use of biomarkers to show sub-cellular effects in meadow voles (Microtus pennsylvanicus) living on an abandoned gold mine site. AB - Run-off from mine tailings ponds constitutes the main anthropogenic release of arsenic in Canada. As a potential consequence, wildlife not normally exposed to arsenic under other circumstances may receive toxicologically relevant concentrations of arsenic compounds in their food and water. To test this hypothesis, and to determine if arsenic is being transported through trophic levels, the arsenic concentrations in members of a short food chain (soil-plant meadow vole) were measured. Arsenic concentrations were higher in exposed organisms compared with those from a reference location. However, elevated concentrations of arsenic do not necessarily indicate impact, and consequently a biomonitoring study was undertaken to determine if there were sub-cellular effects of exposure in meadow voles (Microtus pennsylvanicus) as a consequence of arsenic exposure. In this work, adenosine triphosphate (ATP) and liver glutathione (GSH) levels were used as biomarkers of exposure and the frequency of red blood cell micronuclei (mono- and polychromatic) was used as a biomarker of effect. ATP results were not conclusive but there was a statistically significant relationship between a reduction of GSH in vole livers and increased liver arsenic concentrations. A statistically significant relationship was also observed between increased micronucleated monochromatic red blood cells in voles from arsenic contaminated sites compared to a background location. The results of the GSH and monochromatic red blood cell investigations suggest that there are possible sub-cellular effects on these voles as a consequence of dietary arsenic exposure. This is the first field study in which such effects have been observed in voles living near mine tailings. PMID- 19674773 TI - Correlation of thromboelastography with standard tests of anticoagulation in paediatric patients receiving extracorporeal life support. AB - Children requiring extracorporeal life support (ECLS) are at significant risk for thrombotic and haemorrhagic complications. Thromboelastography (TEG) is increasingly being used to assist in monitoring the coagulation status of critically ill patients. Its role in heparinised children receiving ECLS is unknown. METHODS: A retrospective review of TEG in 27 children (mean age 2 years and 8 months) receiving ECLS in a tertiary paediatric intensive care unit between December 2006 and April 2008. Paired TEG (kaolin and heparinase) analysis was performed on 171 occasions. On all occasions activated partial thromboplastin time (APTT) and platelet count were performed within 4 hours of the TEG (mean 6.5 minutes after TEG). On 158 occasions, the activated clotting time (ACT) was measured simultaneously with TEG. RESULTS: The TEG (kaolin) sample was not interpretable due to the heparin effect in 89 (52%) samples. There was a weak correlation between TEG (heparinase) variables and APTT, and between TEG and ACT with a stronger correlation between TEG (Maximum amplitude) and platelet count. CONCLUSION: TEG monitoring should always include paired samples in heparinised children on ECLS. In this heterogeneous population, weak, and moderate correlations exist between TEG and standard haematological tests. Prospective studies, with simultaneous sampling for TEG and conventional laboratory tests, must be performed in order to establish its absolute utility as a clinical tool in this population. PMID- 19674774 TI - Incidences of kidney, pelvis, ureter, and bladder cancer in a nationwide, population-based cancer registry, Denmark, 1944-2003. AB - OBJECTIVES: To determine changes in the population-based incidence rates, degree of histologic verification, and morphologic distribution of tumors of the kidney, pelvis, and ureter and bladder in both sexes for the duration of 60 years in Denmark. METHODS: Age-standardized incidence rates in 5-year age and calendar intervals, histologic verification and morphologic distribution were derived from reports to the Danish Cancer Registry. RESULTS: The incidence of kidney tumors increased by 78% among men and 32% among women during the study period; the incidence of pelvis and ureter tumors increased by 1070% and 2785%, respectively, and those of bladder tumors increased by 325% and 195%, respectively. The incidence of Wilms tumor was stable; the incidence of other kidney tumors leveled out during the last 30 years of the period and those of other 2 groups during the last 20 years. The frequency of histologic verification was stable for pelvis and ureter cancers and Wilms tumor and increased for kidney and bladder tumors. Most of the kidney tumors were of epithelial origin and most others were of urothelial origin. CONCLUSIONS: The overall incidences of all tumors increased in both sexes during the 60-year period. Some of the increase could be due to improved diagnosis. The recent decline in rates might be due to reduced exposure to risk factors. PMID- 19674775 TI - Pediatric radical abdominal trachelectomy for solitary fibrous tumor of the uterine cervix. AB - OBJECTIVE: Solitary fibrous tumor is a rare mesenchymal tumor initially described arising in the pleura but now reported in many other thoracic and extrathoracic sites. Here we report the first case of a 14-year-old girl who was discovered to have a large solitary fibrous tumor of the uterine cervix, an unusual site for these tumors, to highlight the diagnosis and management of these tumors. METHODS: Case report and review of the literature on extrapleural solitary fibrous tumors. RESULTS: The median age of occurrence is in the late 50's and there is only one previous report in the literature of a solitary fibrous tumor in a patient less than 18 years old. Most patients will be cured by complete surgical resection though late local and distant relapse will occur in a minority of patients, many of whom have indicative histological features. Complete surgical resection of this tumor arising in the uterine cervix, was achieved by performing a radical abdominal trachelectomy. CONCLUSIONS: Review of the literature confirms the good long-term outcome of the majority of these patients with complete surgical resection but emphasizes the importance of close long-term follow-up. PMID- 19674776 TI - Homoeopathy: not a matter for drug-regulatory authorities. PMID- 19674777 TI - Gold nanoparticles with a monolayer of doxorubicin-conjugated amphiphilic block copolymer for tumor-targeted drug delivery. AB - Gold (Au) nanoparticles (NPs) stabilized with a monolayer of folate-conjugated poly(L-aspartate-doxorubicin)-b-poly(ethylene glycol) copolymer (Au-P(LA-DOX)-b PEG-OH/FA) was synthesized as a tumor-targeted drug delivery carrier. The Au-P(LA DOX)-b-PEG-OH/FA NPs consist of an Au core, a hydrophobic poly(l-aspartate doxorubicin) (P(LA-DOX)) inner shell, and a hydrophilic poly(ethylene glycol) and folate-conjugated poly(ethylene glycol) outer shell (PEG-OH/FA). The anticancer drug, doxorubicin (DOX), was covalently conjugated onto the hydrophobic inner shell by acid-cleavable hydrazone linkage. The DOX loading level was determined to be 17 wt%. The Au-P(LA-DOX)-b-PEG-OH/FA NPs formed stable unimolecular micelles in aqueous solution. The size of the Au-P(LA-DOX)-b-PEG-OH/FA micelles were determined as 24-52 and 10-25 nm by dynamic light scattering (DLS) and transmission electron microscopy (TEM), respectively. The conjugated DOX was released from the Au-P(LA-DOX)-b-PEG-OH/FA micelles much more rapidly at pH 5.3 and 6.6 than at pH 7.4, which is a desirable characteristic for tumor-targeted drug delivery. Cellular uptake of the Au-P(LA-DOX)-b-PEG-OH/FA micelles facilitated by the folate-receptor-mediated endocytosis process was higher than that of the micelles without folate. This was consistent with the higher cytotoxicity observed with the Au-P(LA-DOX)-b-PEG-OH/FA micelles against the 4T1 mouse mammary carcinoma cell line. These results suggest that Au-P(LA-DOX)-b-PEG OH/FA NPs could be used as a carrier with pH-triggered drug releasing properties for tumor-targeted drug delivery. PMID- 19674778 TI - Prolonged local antibiotics delivery from hydroxyapatite functionalised with cyclodextrin polymers. AB - Per-operative infection is a common complication for bone-graft surgery. Combining antiseptic agents with graft materials may offer a solution by increasing local drug concentration at target sites. Aiming to achieve a sustained local antibiotic (ATB) delivery for a widely applied bone substitute material - hydroxyapatite (HA), we attempted incorporating hydroxypropyl-beta cyclodextrin polymer (polyHPbetaCD) into microporous HA via impregnating either in a CD monomers mixture solution or a pre-synthesized CD polymer solution, followed by thermal fixation processing. In such functionalised material (CD-HA), polyHPbetaCD could entrap ATBs and release them progressively. Infrared spectroscopic analysis confirmed the presence of polyHPbetaCD in functionalised HA via both processing pathways; polyHPbetaCD functionalisation yields were quantitated by thermogravimetric analysis for optimising the processing regime. Ciprofloxacin (CFX) and vancomycin (VCM), commonly applied in orthopaedics, have been respectively loaded on CD-HA by dip-coating. For both ATBs, kinetic release test in phosphate buffered saline showed significantly increased initial-burst amount and prolonged release from CD-HA compared with those from non functionalised HA. Encouragingly, ATBs loaded CD-HA also revealed a prolonged bacteriostatic activity against Staphylococcus aureus and progressively increased cytocompatibility to osteoblasts (MC3T3-E1). Overall, polyHPbetaCD functionalisation on HA could be an effective drug-delivery model for loading different drug molecules in prevention of infection. PMID- 19674779 TI - Finite element study of scaffold architecture design and culture conditions for tissue engineering. AB - Tissue engineering scaffolds provide temporary mechanical support for tissue regeneration and transfer global mechanical load to mechanical stimuli to cells through its architecture. In this study the interactions between scaffold pore morphology, mechanical stimuli developed at the cell microscopic level, and culture conditions applied at the macroscopic scale are studied on two regular scaffold structures. Gyroid and hexagonal scaffolds of 55% and 70% porosity were modeled in a finite element analysis and were submitted to an inlet fluid flow or compressive strain. A mechanoregulation theory based on scaffold shear strain and fluid shear stress was applied for determining the influence of each structures on the mechanical stimuli on initial conditions. Results indicate that the distribution of shear stress induced by fluid perfusion is very dependent on pore distribution within the scaffold. Gyroid architectures provide a better accessibility of the fluid than hexagonal structures. Based on the mechanoregulation theory, the differentiation process in these structures was more sensitive to inlet fluid flow than axial strain of the scaffold. This study provides a computational approach to determine the mechanical stimuli at the cellular level when cells are cultured in a bioreactor and to relate mechanical stimuli with cell differentiation. PMID- 19674780 TI - An electrochemical glucose biosensor exploiting a polyaniline grafted multiwalled carbon nanotube/perfluorosulfonate ionomer-silica nanocomposite. AB - A glucose biosensor was fabricated with loading of glucose oxidase (GOx) into a new organic-inorganic hybrid nanocomposite. The preparation involves formation of silica network into a Nafion (perfluorosulfonate ionomer) and subsequent loading of polyaniline grafted multiwalled carbon nanotubes (MWNT-g-PANI) onto Nafion silica nanocomposite. Field emission scanning electron microscopy (FE-SEM) of Nafion-silica/MWNT-g-PANI composite reveals the presence of spherical silica particles (sizes in the range 250 nm-1 microm) and tubular MWNT-g-PANI particles. Chronoamperometry and cyclic voltammetry were used to evaluate the performance of biosensor towards glucose. The Nafion-silica/MWCNT-g-PANI/GOx biosensor exhibited a linear response to glucose in the concentration range of 1-10 mm with a correlation coefficient of 0.9972, good sensitivity (5.01 microA/mm), a low response time (approximately 6s), repeatability (R.S.D value of 2.2%) and along term stability. The presence of silica network within Nafion and MWNT-g-PANI synergistically contributes to the performance of the biosensor towards the electrochemical detection of glucose. PMID- 19674781 TI - Bioengineering of a functional sheet of islet cells for the treatment of diabetes mellitus. AB - The present study was designed to establish a novel tissue engineering approach for diabetes mellitus (DM) by fabricating a tissue sheet composed of pancreatic islet cells for in vivo transplantation. Pancreatic islet cell suspensions were obtained from Lewis rats, and plated onto temperature-responsive culture dishes coated with extracellular matrix (ECM) proteins. After the cells reached confluency, islet cells cultured on laminin-5 coated dishes were successfully harvested as a uniformly spread tissue sheet by lowering the culture temperature to 20 degrees C for 20 min. The functional activity of the islet cell sheets was confirmed by histological examination and Insulin secretion assay prior to in vivo transplantation. Histological examination revealed that the harvested islet cell sheet was comprised of insulin- (76%) and glucagon- (19%) positive cells, respectively. In vivo functionality of the islet cell sheet was maintained even 7 days after transplantation into the subcutaneous space of Lewis rats. The present study describes an approach to generate a functional sheet of pancreatic islet cells on laminin-5 coated temperature-responsive dishes, which can be subsequently transplanted in vivo. This study serves as the foundation for the creation of a novel cell-based therapy for DM to provide patients an alternative method. PMID- 19674782 TI - Biomimetic apatite-coated alginate/chitosan microparticles as osteogenic protein carriers. AB - Bone morphogenetic proteins (BMPs) are currently approved for spinal fusion, tibial fracture repair, and maxillofacial bone regeneration. However, BMP pleiotropism, paradoxical activities on precursor cells, and unexpected side effects at local and ectopic sites may limit their usage. Thus, the need remains for alternative osteoinductive factors that provide more bone-specific activities with fewer adverse effects. Nell-1 [Nel-like molecule-1; Nel (a protein highly expressed in neural tissue encoding epidermal growth factor like domain)] is a novel osteogenic protein believed to specifically target cells committed to the osteogenic lineage. The objective of this project is to incorporate Nell-1 into a moldable putty carrier that can adapt to bony defects and deliver Nell-1 to the local microenvironment. We show here that moldability can be achieved by mixing hyaluronan hydrogel with two types of particles: demineralized bone powder for osteoconductivity, and biomimetic apatite-coated alginate/chitosan microparticles for controlled Nell-1 delivery. Besides enhancing overall osteoconductivity of the carrier, the biomimetic apatite coating also provides a more sustained release (approximately 15% cumulative release over 30 days) and greatly reduces the initial burst release that is observed with non-coated alginate/chitosan microparticles (approximately 40% release after 1 day). The efficacy of Nell-1 delivery from these carriers was evaluated in a rat spinal fusion model against Nell-free carriers as controls. At 4 weeks post-implantation, Nell-1 enhanced spinal fusion rates as assessed by manual palpation, radiographs, high-resolution micro-computerized tomography (microCT), and histology. This moldable putty carrier system appears to be a suitable carrier for promoting osteogenesis, and will be further evaluated in larger animal models over longer periods to follow the remodeling of the regenerated bone. PMID- 19674783 TI - Development and analysis of multi-layer scaffolds for tissue engineering. AB - The development of 3D scaffolds consisting of stacked multi-layered porous sheets featuring microchannels is proposed and investigated in this work. In this concept, the inner-porosity of the sheets allows diffusion of nutrients and signalling products between the layers whereas the microchannels facilitate nutrient supply on all layers as they provide space for the culture medium to be perfused throughout the scaffold. Besides the above, these scaffolds have excellent distribution of the cells as seeding and attaching of the cells occurs on individual layers that are subsequently stacked. In addition, these scaffolds enable gaining local data from within the scaffolds as unstacking of the stacked layers allows for determination of various parameters per layer. Here, we show the proof of this concept by culturing C2C12 pre-myoblasts and A4-4 cells on stacked Poly(l-lactic acid) (PLLA) sheets featuring microchannels. The results obtained for culturing under static conditions clearly indicate that despite inhibited cell proliferation due to nutrient limitations, diffusion between the layers takes place and cells on various layers stay viable and also affect each other. Under dynamic conditions, medium flow through the channels improves nutrient availability to the cells on the various layers, drastically increasing cell proliferation on all layers. PMID- 19674784 TI - Poly(ethylene glycol) hydrogels formed by thiol-ene photopolymerization for enzyme-responsive protein delivery. AB - Degradable hydrogels have been extensively used in biomedical applications such as drug delivery, and recent interest has grown in hydrogels that degrade in recognition of a cellular response. This contribution describes a poly(ethylene glycol) (PEG) hydrogel platform with human neutrophil elastase (HNE) sensitive peptide cross-links formed using thiol-ene photopolymerization rendering the gel degradable at sites of inflammation. Further, protein therapeutics can be physically entrapped within the network and selectively released upon exposure to HNE. HNE-responsive hydrogels exhibited surface erosion where the degradation kinetics was influenced by changes in peptide k(cat), concentration of HNE, and concentration of peptide within the gel. Using this platform, we were able to achieve controlled, zero-order release of bovine serum albumin (BSA) in the presence of HNE, and release was arrested in the absence of HNE. To further exploit the advantages of surface eroding delivery systems, a smaller protein (carbonic anhydrase) was delivered at the same rate as BSA and only dependent on gel formulation and environmental conditions. Also, protein release was predicted from a 3-layered hydrogel device using mass loss data. Lastly, the bioactivity of lysozyme was maintained above 90% following the exposure to thiol-ene photopolymerization conditions. PMID- 19674785 TI - The growth and differentiation of mesenchymal stem and progenitor cells cultured on aligned collagen matrices. AB - Cell-matrix interactions are paramount for the successful repair and regeneration of damaged and diseased tissue. Since many tissues have an anisotropic architecture, it has been proposed that aligned extracellular matrix (ECM) structures in particular could guide and support the differentiation of resident mesenchymal stem and progenitor cells (MSCs). We therefore created aligned collagen type I structures using a microfluidic set-up with the aim to assess their impact on MSC growth and differentiation. In addition, we refined our aligned collagen matrices by incorporating the glycosaminoglycan (GAG) heparin to demonstrate the versatility of the applied methodology to study multiple ECM components in a single system. Our reconstituted, aligned ECM structures maintained and allowed multilineage (osteogenic/adipogenic/chondrogenic) differentiation of MSCs. Most noticeable was the observation that during osteogenesis, aligned collagen substrates choreographed ordered matrix mineralization. Likewise, myotube assembly of C2C12 cells was profoundly influenced by aligned topographic features resulting in enhanced myotube organization and length. Our results shed light on the regulation of MSCs through directional ECM structures and demonstrate the versatility of these cell culture platforms for guiding the morphogenesis of tissue types with anisotropic structures. PMID- 19674786 TI - Placentation in the Egyptian slit-faced bat Nycteris thebaica (Chiroptera: Nycteridae). AB - Bats are a highly successful, widely distributed group, with considerable variation in placental structure. The Egyptian slit-faced bat Nycteris thebaica is a member of one of the few families with previously undescribed placentation. It was found that, although the interhemal type of the Nycteris placenta is endotheliochorial with a single layer of cytotrophoblast, the arborizing pattern of the maternal vessels and especially the extraordinary major placental artery differs from the placenta of the emballonurid bats to which this family is considered to be most closely related. The major placental artery providing maternal blood to the vessels of the placental disk has a highly glycosylated matrix surrounded by two-layered folds of trophoblast, forming an apparently rigid structure of unique morphology. The yolk sac is collapsed, with hypertrophied endodermal and mesothelial cells similar to many other bat species. The paraplacenta is extensive with abundant fetal vessels underlying cytotrophoblast and syncytial trophoblast layers, fronting on an endometrium that largely lacks uterine epithelial cells but has large decidual cells and is poorly vascularized. The placenta of Nycteris lacks a hemophagous region, unlike the emballonurid bats Taphozous and Saccopteryx. Although the latter two species have similar placentas, the placental structure of Nycteris does little to relate it to the other family within the Emballonuroidea. Shared and divergent reproductive characters are discussed in relationship to bat phylogenetic relationships. PMID- 19674787 TI - A placental protective role for trophoblast-derived TNF-related apoptosis inducing ligand (TRAIL). AB - Recent studies show that apoptosis, programmed cell death, plays an important role in the normal development of the human placenta and that an altered balance between proliferation and apoptosis of villous trophoblasts is associated with abnormal pregnancies. The TNF-related apoptosis-inducing ligand (TRAIL) is a molecule belonging to TNF superfamily. The role of TRAIL and its Death Receptor 5 (DR5) in regulating villous trophoblast cell turnover in normal and pathologic pregnancies remains to be explored. In order to elucidate the role of TRAIL in the regulation of placental growth, primary cytotrophoblast cells were isolated from normal term placentas (n=13) and cultured for 18 and 66h to generate mononucleate and multinucleate trophoblasts, respectively. The protein expression and localisation of TRAIL and DR5 were determined by Western blotting and immunofluorescence. Secreted sTRAIL was also measured by ELISA. Trophoblast apoptosis was measured by TUNEL in the presence of recombinant TRAIL (rTRAIL), and DR5 relocalisation was assessed by immunostaining after 18h exposure to TNFalpha. We demonstrated that TRAIL protein expression and the secretion of soluble TRAIL (sTRAIL) were down-regulated in syncytialised villous trophoblasts and that sTRAIL was independent of biochemical differentiation, as TRAIL neutralizing antibody (2E5) failed to influence hCG production. TRAIL immunoreactivity was detected in mono- and multinucleated trophoblast cells and localised to the cytoplasm and cellular membranes -- more intense staining was associated with apoptotic nuclei. rTRAIL failed to induce apoptosis in trophoblasts cells owing to the nuclear localisation of DR5. However, TNFalpha treatment caused the redistribution of intracellular DR5 to the cell surface, potentiating apoptotic susceptibly to exogenously administered rTRAIL. These findings highlight a mechanism by which TRAIL and DR5 serve to protective trophoblasts in normal development, but may be activated in conditions of excessive TNFalpha. PMID- 19674788 TI - Acute myeloid leukemia and Langerhans' cell histiocytosis: multiple theories for an unusual presentation. PMID- 19674789 TI - An open-label, Phase I study of cediranib (RECENTIN) in patients with acute myeloid leukemia. AB - VEGFR and c-Kit signaling pathways may contribute to the pathophysiology of acute myeloid leukemia (AML). Thirty-five patients with AML received cediranib (RECENTIN), an oral, highly potent VEGF signaling inhibitor with c-Kit activity, at doses of < or =30 mg/day. The most common adverse events were diarrhea, hypertension and fatigue. Six patients experienced an objective response (3 each at 20 and 30 mg). Dose- and time-dependent reductions in sVEGFR-2 were observed, and there was a positive correlation between cediranib exposure and the change in plasma VEGF levels from baseline. Cediranib was generally well tolerated and showed preliminary evidence of activity as a monotherapy. PMID- 19674790 TI - Bortezomib-induced peripheral neuropathy in multiple myeloma: a comparison between previously treated and untreated patients. AB - Peripheral neuropathy (PN), with neuropathic pain as main symptom, represents the dose-limiting toxicity of the proteasome inhibitor bortezomib. Aim of this study was to compare the incidence, risk factors, severity and outcome of PN and neuropathic pain in patient treated with bortezomib up-front or at relapse. We studied 55 patients with multiple myeloma (MM) who received bortezomib as first line therapy and 70 pre-treated patients who received bortezomib in relapse or progression. Regarding PN, no differences were found among untreated and pre treated patients in the incidence (55% vs 52%, p=0.43), severity (NCI grade 3-4 9% vs 14%, p=0.27), and outcome (improved/resolved 90% vs 91%, p=0.58). Concerning neuropathic pain, the incidence was lower (50% vs 81%, p=0.008) and solved earlier (35 days vs 91 days, p=0.02) in untreated compared with pre treated patients. Untreated patients needed dose modification less frequently (36% vs 73%, p=0.012). No correlation was found between development of PN and prior exposure to potentially neurotoxic drugs such as thalidomide, vincristine, and cysplatin. Age represented the main risk factor for PN (p=0.036) with an increase in risk of PN amounting to 6% per year of age. In conclusion, incidence, severity and outcome of bortezomib-related PN are similar in untreated and pre treated MM patients except for neuropathic pain which has lower incidence and shorter duration in untreated patients with less frequent need for bortezomib discontinuation. Age emerges as the most relevant risk factor for peripheral neuropathy, with a risk increase for PN of 6% per year of age. PMID- 19674791 TI - Interictal arterial spin-labeling MRI perfusion in intractable epilepsy. AB - INTRODUCTION: Magnetic resonance imaging (MRI) is required for the investigation of surgically intractable epilepsy. In addition to the standard MRI techniques, perfusion sequences can be added to improve visualization of underlying pathological changes. Arterial spin-labeling (ASL) MRI perfusion does not require contrast administration and, for this reason, may have advantages in these patients. METHODS: We report here on 16 patients with epilepsy who underwent MRI of the brain with ASL and positron emission tomography (PET). RESULTS: Despite a slightly reduced resolution with ASL, we found a correlation between ASL, PET and electrophysiological data, with hypoperfusion on ASL that corresponded with hypoperfusion on interictal PET. CONCLUSION: Given the correlation between ASL and PET and electrophysiology, perfusion with ASL could become part of the standard work-up in patients with epilepsy. PMID- 19674792 TI - Interactions of histidine-rich glycoprotein with immunoglobulins and proteins of the complement system. AB - This study describes how the serum protein histidine-rich glycoprotein (HRG) affects the complement system. We show that HRG binds strongly to several complement proteins: C1q, factor H and C4b-binding protein and that it is found complexed with these proteins in human sera and synovial fluids of rheumatoid arthritis patients. HRG also binds C8 and to a lesser extent mannose-binding lectin, C4 and C3. However, HRG alone neither activates nor inhibits complement. Both HRG and C1q bind to necrotic cells and increase their phagocytosis. We found that C1q competes weakly with HRG for binding to necrotic cells whilst HRG does not compete with C1q. Furthermore, HRG enhances complement activation on necrotic cells measured as deposition of C3b. We show that HRG inhibits the formation of immune complexes of ovalbumin/anti-ovalbumin, whilst the reverse holds for C1q. Immune complexes formed in the presence of HRG show enhanced complement activation, whilst those formed in the presence of C1q show diminished complement activation. Taken together, HRG may assist in the maintenance of normal immune function by mediating the clearance of necrotic material, inhibiting the formation of insoluble immune complexes and enhancing their ability to activate complement, resulting in faster clearance. PMID- 19674793 TI - Investigating motionese: The effect of infant-directed action on infants' attention and object exploration. AB - Adults modify their communication when interacting with infants, and these modifications have been tied to infant attention. However, the effect infant directed action on infant behavior is understudied. This study examined whether infant-directed action affects infants, specifically their attention to and exploratory behaviors with objects. Forty-eight 8- to 10-month-old infants and their caregivers participated in a laboratory session during which caregivers demonstrated objects to infants using infant-directed action. Results indicated that variation in amplitude and repetition were tied to differences in infant attention, and varying levels of repetition were tied to differences in object exploration. PMID- 19674794 TI - Canadian Network for Mood and Anxiety Treatments (CANMAT) clinical guidelines for the management of major depressive disorder in adults. III. Pharmacotherapy. AB - BACKGROUND: In 2001, the Canadian Psychiatric Association and the Canadian Network for Mood and Anxiety Treatments (CANMAT) partnered to produce evidence based clinical guidelines for the treatment of depressive disorders. A revision of these guidelines was undertaken by CANMAT in 2008-2009 to reflect advances in the field. METHODS: The CANMAT guidelines are based on a question-answer format to enhance accessibility to clinicians. An evidence-based format was used with updated systematic reviews of the literature and recommendations were graded according to Level of Evidence using pre-defined criteria. Lines of Treatment were identified based on criteria that included Levels of Evidence and expert clinical support. This section on "Pharmacotherapy" is one of 5 guideline articles. RESULTS: Despite emerging data on efficacy and tolerability differences amongst newer antidepressants, variability in patient response precludes identification of specific first choice medications for all patients. All second generation antidepressants have Level 1 evidence to support efficacy and tolerability and most are considered first-line treatments for MDD. First generation tricyclic and monoamine oxidase inhibitor antidepressants are not the focus of these guidelines but generally are considered second- or third-line treatments. For inadequate or incomplete response, there is Level 1 evidence for switching strategies and for add-on strategies including lithium and atypical antipsychotics. LIMITATIONS: Most of the evidence is based on trials for registration and may not reflect real-world effectiveness. CONCLUSIONS: Second generation antidepressants are safe, effective and well tolerated treatments for MDD in adults. Evidence-based switching and add-on strategies can be used to optimize response in MDD that is inadequately responsive to monotherapy. PMID- 19674795 TI - Shared genetic factors in the co-occurrence of symptoms of depression and cardiovascular risk factors. AB - BACKGROUND: We aim to investigate the extent to which shared genetic and shared environmental factors play a role in the co-occurrence of symptoms of depression and cardiovascular risk factors. METHODS: The analyses included 2383 individuals from a genetically isolated population in the Netherlands (mean age 48.7 years (standard deviation 15.1), percentage of women 56.9%). Symptoms of depression were assessed using the Center for Epidemiology Studies Depression Scale (CES-D) and the Depression subscale of the Hospital Anxiety and Depression Scale (HADS D). Assessment of cardiovascular risk factors included systolic and diastolic blood pressure, plasma, glucose levels, high and low density lipoprotein (HDL, LDL) and total cholesterol levels. RESULTS: Overall, we found that HADS-D was significantly correlated to total cholesterol levels (correlation coefficient [rho]=0.05), and inversely associated to HDL (rho=-0.06). Statistically significant genetic correlations (rho(G)) were found between CES-D scores and total plasma cholesterol (rho(G)=0.30), LDL (rho(G)=0.31) and total cholesterol/HDL ratios (rho(G)=0.25). For HADS-D scores, a significant genetic correlation was found with total cholesterol/HDL ratios (rho(G)=0.27). Environmental correlations (rho(E)) with an opposite direction were found between CES-D and both total cholesterol (rho(E)=-0.16) and LDL (rho(E)=-0.15). LIMITATION: By adjusting for sibship, we are taking into account environmental effects, however we cannot exclude dominance variance. CONCLUSIONS: Our study shows that there is evidence for shared genetic factors contributing to the co occurrence of symptoms of depression and lipid levels. This finding suggests a joint genetic pathogenesis. Future research is encouraged to assess susceptibility genes for mood disorders to be studied for cardiovascular disorders and vice versa. PMID- 19674796 TI - Canadian Network for Mood and Anxiety Treatments (CANMAT) clinical guidelines for the management of major depressive disorder in adults. I. Classification, burden and principles of management. AB - BACKGROUND: Major depressive disorder (MDD) is one of the most burdensome illnesses in Canada. The purpose of this introductory section of the 2009 revised CANMAT guidelines is to provide definitions of the depressive disorders (with an emphasis on MDD), summarize Canadian data concerning their epidemiology and describe overarching principles of managing these conditions. This section on "Classification, Burden and Principles of Management" is one of 5 guideline articles in the 2009 CANMAT guidelines. METHODS: The CANMAT guidelines are based on a question-answer format to enhance accessibility to clinicians. An evidence based format was used with updated systematic reviews of the literature and recommendations were graded according to the Level of Evidence using pre-defined criteria. Lines of Treatment were identified based on criteria that included evidence and expert clinical support. RESULTS: Epidemiologic data indicate that MDD afflicts 11% of Canadians at some time in their lives, and approximately 4% during any given year. MDD has a detrimental impact on overall health, role functioning and quality of life. Detection of MDD, accurate diagnosis and provision of evidence-based treatment are challenging tasks for both clinicians and for the health systems in which they work. LIMITATIONS: Epidemiologic and clinical data cannot be seamlessly linked due to heterogeneity of syndromes within the population. CONCLUSIONS: In the eight years since the last CANMAT Guidelines for Treatment of Depressive Disorders were published, progress has been made in understanding the epidemiology and treatment of these disorders. Evidence supporting specific therapeutic interventions is summarized and evaluated in subsequent sections. PMID- 19674797 TI - Immunoglobulins and transcription factors in adenoids of children with otitis media with effusion and chronic rhinosinusitis. AB - OBJECTIVE: To evaluate the level of immunoglobulins and expression of the transcription factors in the adenoids. METHODS: The study population consisted of 30 children with otitis media with effusion (OME), 22 children with chronic rhinosinusitis (CRS) and 27 children with adenoid hyperplasia. IgA, IgG, IgD and IgM concentrations in these three groups were compared immunohistochemically. We also compared the levels of expression of B lymphocyte inducer of maturation program 1 (BLIMP-1), a promoter of plasmacytosis, and B cell leukemia/lymphoma-6 (BCL-6), a repressor of plasmacytosis, in the adenoids of these children. RESULTS: The expression of antibody to Ig A in the OME and CRS groups each was significantly lower than the score in the adenoid hyperplasia group. The staining scores of antibodies to IgG, IgD and Ig M did not differ significantly among the three groups. The expression of antibody to BLIMP-1 in the CRS and adenoid hyperplasia groups showed a significant difference. Staining scores of Antibody to BCL-6 did not differ significantly among the three groups. CONCLUSIONS: We found that the reduction of IgA in the adenoids was associated with pediatric OME and CRS, and that reduction of expression of the transcription factor BLIMP-1 rather than BCL-6 was associated with CRS. PMID- 19674799 TI - Comparing the inhibitory effects of five protoxicant organophosphates (azinphos methyl, parathion-methyl, chlorpyriphos-methyl, methamidophos and diazinon) on the spontaneously beating auricle of Sparus aurata: an in vitro study. AB - Organophosphates (OPs) can provoke toxicity by inhibiting acetylcholinesterase (AChE) in non-target organisms, like fish. In a previous pilot study, the anticholinesterase effects of paraoxon on the heart of Sparus aurata were examined [Tryfonos, M., Antonopoulou, E., Papaefthimiou, C., Chaleplis, G., Theophilidis, G., 2009. An in vitro assay for the assessment of the effects of an organophosphate, paraoxon, and a triazine, atrazine, on the heart of the gilthead sea bream (Sparus aurata). Pest. Biochem. Physiol. 93, 40-46]. The objective of the present study was to investigate the effects of the five protoxicant OPs, azinphos-methyl (MeAZP), parathion-methyl (MePS), chlorpyriphos-methyl (MeCCP), methamidophos (MET) and diazinon (DZ), on the spontaneously beating auricle of S. aurata. The results showed that: (1) MeAZP and MET induced exclusively cholinergic effects on auricle contractility. These effects were expressed as a significant decrease in the force and frequency of contractions and were fully reversible (140%) after the application of the muscarinic cholinergic receptor antagonist, atropine (15 microM). MeAZP was found to be the most effective anticholinesterase compound, with an IC(50) of 2.19+/-1.05 microM (n=6), while MET was less effective, with an IC(50) of 72.3+/-1.2 microM (n=6). (2) DZ and MePS, although classified as OPs, induced non-cholinergic effects. These effects were observed as an irreversible decrease in force and frequency of the auricle in all the concentrations examined; the depression is retained even after application of 15 microM atropine. (3) MeCCP was halfway between a typical OP and an OP lacking anticholinesterase properties, since there was a partial recovery in the force, but no recovery in the frequency of the auricle contractions. (4) The toxicity order, based on the IC(50), was as follows: MeAZP, 2.19+/-1.05 microM>paraoxon, 3.2+/-1.5 microM"MET, 72.3+/-1.2 microM>MePS, 80.3+/-1.03 microM>MeCPP, 93.7+/-1.01 microM>DZ, 164+/-1.01 microM. (5) There was a good correlation (r=0.779, p=0.04, n=5) between IC(50) and the previously determined logP (octanol:water partition coefficient) values for MeAZP, paraoxon, MeCCP, MePS and DZ. The results indicated that the increase in lipophilicity of MePS, MeCCP and DZ is accompanied by a decrease in their acute cardiotoxic properties in vitro. The non-cholinergic effects of these relatively high lipophilic OPs, might be caused by their tendency to distribute preferentially in the lipid bilayer of cardiac cells, affecting the proper functioning of the ionic channels which regulate the force (Ca(2+) channels) and the frequency (K(+) channels) of the spontaneous auricle contractions. PMID- 19674798 TI - Parent versus child assessment of quality of life in children using cochlear implants. AB - OBJECTIVE: Children with hearing loss who use cochlear implants have lower quality of life (QoL) in social situations and lower self-esteem than hearing peers. The child's QoL has been assessed primarily by asking the parent rather than asking the child. This poses a problem because parents have difficulty judging less observable aspects like self-esteem and socio-emotional functioning, the domains most affected by hearing loss. METHODS: This case-control study evaluated QoL in 50 preschoolers using a cochlear implant and their parents with the Kiddy KINDL((r)), an established QoL measure. Children's responses were compared to a hearing control group and correlated with demographic variables. We used a questionnaire for parents and a face-to-face interview with children. T tests were used to compare (a) paired parent-child ratings and (b) children with cochlear implants versus normal hearing. Pearson rank correlations were used to compare QoL with demographic variables. RESULTS: Children using cochlear implants rated overall QoL significantly more positively than their parents (M(Difference)=4.22, p=.03). Child rating of QoL did not differ significantly by auditory status (cochlear implant (82.8) vs. hearing (80.8), p=.42). Overall QoL correlated inversely with cochlear implant experience and chronologic age, but did not correlate with implantation age. CONCLUSIONS: Preschool children using cochlear implants can assess adequately their own QoL, but parents afford valuable complementary perspective on the child's socio-emotional and physical well-being. Preschool children using cochlear implants rate overall QoL measures similar to hearing peers. A constellation of QoL measures should be collected to yield a better understanding of general QoL as well as specific domains centered on hearing loss. PMID- 19674800 TI - Impact of elastic stall partitions on tied dairy cows' behaviour and stall cleanliness. AB - The aim of this study was to investigate the effects of soft elastic stall partitions on the positions in own or neighbouring stalls by dairy cows, faeces and urine, and of contact between animal and partition. There were two designs of a soft elastic partition, here called I-partition and lambda-partition. The study was performed on a commercial dairy herd where the partitions had been developed. After clinical examination 16 lactating cows were selected and used as focal animals, and paired for video recordings. The animals were placed, according to their size, on stalls with different breadths and lengths. Each pair of focal animals was video recorded continuously for 9 days according to the following sequence: (1) with partition 3x24 h; (2) without partition 3x24 h; (3) with partition again 3x24 h. Before each such session each focal pair was again clinically examined. Behaviours were analysed per hour, summarised per 24h and analysed with a mixed linear model or Wilcoxon Rank Sum test. Without partitions more urine (p<0.05) and faeces (p<0.05) landed on the neighbouring cow's stall and the time standing 45 degrees to the manger was longer (p<0.01), whereas the time standing 90 degrees to the manger was shorter (p<0.01). Without partitions three out of the 16 cows were standing and three were lying parallel to the manger. Presence or absence of partitions did not influence the total lying time per 24 h. Cows with the lambda-partition defecated more seldom in neighbouring stalls than cows with the I-partition (p<0.01). It is concluded that without partitions between tied animals more eliminations on neighbouring stalls and more deviations from the 90 degrees position in relation to the manger occurred. This seems to be the first experimental study of how partitions influence the behaviour of tied cattle and the results support the general advice that tied cattle should have partitions. PMID- 19674802 TI - When 1+1=1: the unification of independent actors revealed through joint Simon effects in crossed and uncrossed effector conditions. AB - The "Simon effect" describes a pattern of reaction times (RTs) where responses to symbolic information are shorter when the information is presented on the same side of space as the desired response than when it is on the opposite side of space. For example, if right hand responses are required for green targets and left hand responses for red targets, RTs with the right hand are shorter when the green target appears on the right side than on the left side. It has been reported that Simon effects also appear when two individuals perform independent components of a Simon effect task. It has been suggested that such joint Simon effects occur because participants represent the action of their partner. It is unclear, however, if the joint Simon effect emerges because: (1) each partner represents the other's action; (2) each partner is using the other person or their response as an environmental reference; or (3) an intra-hemispheric processing advantage due to the lateralized cerebral organization of perceptual and motor systems. The present study distinguished between these possibilities by asking pairs of participants to perform in conditions in which they crossed their arms into the other person's space. Consistent with within-person Simon effects, joint Simon effects were observed in uncrossed- and crossed limb conditions. These results support a response co-representation explanation of joint Simon effects. It is suggested that the processes underlying the evoked representations have developed to allow two independent agents to form temporary synergies to facilitate efficient task completion. PMID- 19674803 TI - Evaluation of the "testing and scheduling" strategy for control of Campylobacter in broiler meat in The Netherlands. AB - "Testing and scheduling" has been proposed as a strategy for control of Campylobacter in broiler meat. By this strategy, flocks with high numbers of Campylobacter in fecal samples would be diverted away from fresh meat production at the entrance of the broiler meat processing plant. Risk assessment studies suggest that this would effectively decrease human health risks, if these flocks are responsible for the meat products with the highest Campylobacter numbers. To investigate the effect of this control strategy, the numbers of Campylobacter were determined in fecal samples from transport containers, and in cecal and breast meat samples from birds in 62 broiler chicken flocks. Results from direct plating and enrichment were combined by a statistical method that allows the inclusion of censored data. As the implementation of "testing and scheduling" requires a rapid on-site test to detect high numbers of Campylobacter, a lateral flow immuno-assay (LFA) was developed and applied to the fecal samples collected from containers. The Campylobacter prevalence in broiler flocks in the autumn of 2007 was found to be 85.4% by traditional microbiological methods. Campylobacter could be isolated from breast meat samples from 42% of the flocks. There was limited agreement between Campylobacter results for the three types of samples and weak correlation between the quantitative results for fecal or cecal samples and meat samples. Agreement between the results of LFA and traditional methods was poor. These findings do not support the implementation of "testing and scheduling" as a practical control strategy, because of both measurement uncertainties and shortcomings in understanding the dynamics of transmission and survival of Campylobacter in the broiler meat processing plant. The limited correlation between Campylobacter contamination of cecal samples and breast meat samples, as observed in this study, suggests that cecal samples are no good indicator for human exposure to Campylobacter. PMID- 19674801 TI - Novel opportunities for computational biology and sociology in drug discovery. AB - Current drug discovery is impossible without sophisticated modeling and computation. In this review we outline previous advances in computational biology and, by tracing the steps involved in pharmaceutical development, explore a range of novel, high-value opportunities for computational innovation in modeling the biological process of disease and the social process of drug discovery. These opportunities include text mining for new drug leads, modeling molecular pathways and predicting the efficacy of drug cocktails, analyzing genetic overlap between diseases and predicting alternative drug use. Computation can also be used to model research teams and innovative regions and to estimate the value of academy industry links for scientific and human benefit. Attention to these opportunities could promise punctuated advance and will complement the well-established computational work on which drug discovery currently relies. PMID- 19674804 TI - Phytate degradation by human gut isolated Bifidobacterium pseudocatenulatum ATCC27919 and its probiotic potential. AB - The growing awareness of the relationship between diet and health has led to an increasing demand for food products that support health above and beyond providing basic nutrition. Probiotics are live organisms present in foods, which yield health benefits related to their interactions with the gastrointestinal tract. Phytases are a subgroup of phosphatases that catalyse the desphosphorylation of phytate, which reduces its negative impact on mineral bioavailability, and generates lower inositol phosphates. The aims of this investigation were to (i) study the ability of the probiotic candidate Bifidobacterium pseudocatenulatum to degrade phytate in synthetic medium, to (ii) identify the lower inositol phosphates generated, to (iii) study its survival under conditions mimicking gastrointestinal passage and finally to (iv) assess adhesion of the bacteria to Caco-2 cells. The first steps of InsP(6) degradation by B. pseudocatenulatum phytate-degrading enzyme/s were preferentially initiated at the DL-6-position and 5-position of the myo-inositol ring. It suggests that the main InsP(6) degradation pathway by B. pseudocatenulatum by sequential removal of phosphate groups was D/L-Ins(1,2,3,4,5)P(5) or D/L-Ins(1,2,3,4,6)P(5); D/L-Ins(1,2,3,4)P(4); to finally Ins(1,2,3)P(3) and D/L-Ins(1,2,4)P(3)/D/L Ins(1,3,4)P(3). This human strain also showed a notable tolerance to bile as well as a selective adhesion capacity (adhesion to control surfaces was zero), to human intestinal Caco-2 cells comparable to the commercial probiotic B. lactis. The phytate-degrading activity constitutes a novel metabolic trait which could contribute to the improvement of mineral absorption in the intestine as a nutritional probiotic feature with potential trophic effect in human gut. PMID- 19674805 TI - Serum lipid and hsCRP levels in prediabetes--impaired fasting glucose (IFG) and impaired glucose tolerance (IGT). AB - AIM: To evaluate cardiovascular risk in prediabetes (IFG and IGT) assessed by serum lipid and hsCRP levels. SUBJECTS AND METHODS: 445 subjects with prediabetes (248 with IFG, 197 with IGT), 318 patients with newly-diagnosed diabetes (NDD) and a group of 477 age- and BMI-matched subjects with normal glucose tolerance (NGT) were enrolled. Glucose tolerance was studied during oral glucose tolerance test (OGTT) and 2006 WHO criteria were applied. Serum hsCRP and lipids (total cholesterol, triglycerides, HDL-cholesterol (HDL-c) and free fatty acids (FFAs) were measured. RESULTS: Both IFG and IGT showed significantly atherogenic changes in serum lipid and hsCRP levels when compared to NGT. Subjects with IGT presented with significantly higher triglycerides (p=0.01) and FFAs (p<0.0001) and significantly lower HDL-c (p=0.04) as compared to IFG. IFG showed significantly higher levels of HDL-c (p<0.0001) and lower levels of triglycerides (p<0.0001), FFAs (p<0.001) and hsCRP (p=0.04) as compared to NDD, while IGT differed from NDD only in the lower hsCRP (p=0.04). CONCLUSIONS: Both IFG and IGT are associated with increased cardiovascular risk as assessed by serum lipid and hsCRP levels. The risk is different in the two categories of prediabetes, IGT being characterized by a more atherogenic risk profile, similar to that in NDD. PMID- 19674806 TI - Effect of rosiglitazone, metformin and medical nutrition treatment on arterial stiffness, serum MMP-9 and MCP-1 levels in drug naive type 2 diabetic patients. AB - The aim of the study was to evaluate the long-term effect of rosiglitazone and metformin monotherapy with medical nutrition treatment (MNT) and of MNT alone on arterial stiffness, serum monocyte chemoattractant protein (MCP)-1 and matrix metalloproteinase (MMP)-9 in drug naive patients with type 2 diabetes mellitus. Fifty type 2 diabetic patients were randomized to receive rosiglitazone 4 mg/day (n=19) or metformin 850 mg/day (n=16) with MNT or MNT alone (n=15), for 52 weeks. Arterial stiffness was assessed by using large and small artery elasticity index (SAEI and LAEI, respectively). SAEI, LAEI, serum MCP-1 and MMP-9 levels were measured at baseline and following 52 weeks of treatment. SAEI was improved only in the rosiglitazone group, and the difference was still statistically significant when the three groups were compared (p=0.024). There were no differences in LAEI in inter- and intragroup comparisons at the end of the study. Serum MMP-9 levels were decreased in the metformin (-13.5+/-34.8%, p=0.02) and rosiglitazone (-27.2+/-51.0%, p=0.023) groups compared with baseline values, whereas no significant change was seen in serum MCP-1 levels. These results suggest that rosiglitazone monotherapy has favorable effects on arterial stiffness compared with metformin monotherapy independent of glycemic control. PMID- 19674807 TI - Swinnen and DeVries: "Higher dose requirements with insulin detemir in type 2 diabetes-three cases and a review of the literature" [Diab. Res. Clin. Pract. 84 (May (2)) (2009) e24-6 (Epub 2009 February 28)]. PMID- 19674808 TI - The impact of family history of type 2 diabetes on pancreatic beta-cell function. AB - AIMS: To study the impact of genetic factor on pancreatic beta-cell function in the Chinese population. METHODS: 233 first-degree relatives of patients with type 2 diabetes (T2D) with no history of blood glucose abnormalities and their 190 spouses, who did not have a family history of T2D, underwent a 75-g oral glucose tolerance test (OGTT). Based upon the OGTT, these two groups were further divided into three subgroups, including groups with normal glucose tolerance (NGT), impaired glucose regulation (IGR), and type 2 diabetes. Insulin resistance (IR) was evaluated using the homeostasis model assessment-IR (HOMA-IR), beta-cell function indices of basal and first-phase were measured by DI1 (HOMA-beta/HOMA IR) and DI2 (DeltaI30/DeltaG30/HOMA-IR), respectively. RESULTS: Among the first degree relatives and their spouses, the HOMA-IR was highest in the T2D group and lowest in the NGT group. However, the HOMA-beta, DI1 and DI2 declined significantly with progressive reductions in glucose tolerance (P<0.01 or 0.05). DI1 and DI2 of the NGT group of first-degree relatives (FNGT) were significantly lower than those of the spouse NGT (SNGT) group (P<0.05). DI1 and DI2 of the IGR of first-degree relatives (FIGR) group were significantly lower than those of the spouse IGR (SIGR) group. CONCLUSIONS: Defects in pancreatic beta-cell function exist in the first-degree relatives, who have different glucose tolerance statuses, of T2D patients. These defects are more profound in FNGT and FIGR when compared to their spouses in corresponding glucose tolerance subgroups. However, there is no difference in IR between the corresponding glucose tolerance subgroups of the first-degree relatives and their spouses. It suggests that the genetic factor possibly aggravates beta-cell lesion. PMID- 19674809 TI - A tutorial on discrete-event simulation for health policy design and decision making: optimizing pediatric ultrasound screening for hip dysplasia as an illustration. AB - BACKGROUND: It is increasingly recognized that healthcare is a complex system with limited resources and many interacting sources of both positive and negative feedback. Discrete-event simulation (DES) is a tool that readily accommodates questions of capacity planning, throughput management and interacting resources. As a result the use of DES in informing healthcare decision making is increasing. However, understanding when and how to build a DES model and use it for policy making is not yet a common knowledge. METHODS: The steps in building a DES model will be demonstrated using a real-world example, i.e., pediatric ultrasound screening for hip dysplasia. The main components of a DES model such as entities, resources and queues will be introduced and we will examine questions such as referral schedule, number of ultrasound machines and type of screeners and how these entities interact. Finally a review of the statistical techniques appropriate to DES will be provided. CONCLUSION: Discrete-event simulation is a valuable tool in the policymakers armentarium. It can be used effectively to analyze and understand complex healthcare systems and policy problems such as population screening. PMID- 19674810 TI - Cesarean section deliveries in the occupied Palestinian territory (oPt): an analysis of the 2006 Palestinian Family Health Survey. AB - OBJECTIVE: Against the backdrop of a rise in cesarean section deliveries from 6.0% in 1996 to 14.8% in 2006, the objective of this study was to investigate socio-demographic, clinical and service-related factors associated with cesarean sections in the occupied Palestinian territory. METHODS: Data from the Palestinian Family Health Survey 2006 were used to examine last births in the 5 years preceding the survey to women aged 15-49 years. Bivariate and multivariate associations between type of delivery (dependent variable) and selected factors were analyzed using logistic regression. Selected maternal outcomes were also investigated with type of delivery as the independent variable. RESULTS: Cesarean section deliveries were significantly associated with maternal age (35+ years), primiparity, low birth weight and residence area in the West Bank and Gaza. There was no significant difference in the prevalence of cesarean deliveries by sector in the West Bank, but in Gaza, they were significantly more common in the governmental sector. CONCLUSIONS: There is a need for detailed audits of cesarean section deliveries, nationally and at the facility level, in order to avoid unnecessary interventions in the context of high fertility, rising poverty and fragmented health services. Variations by governorate should be studied further for focused interventions. PMID- 19674811 TI - The relationship between response to previous systemic treatment and the efficacy of subsequent pemetrexed therapy in advanced non-small cell lung cancer. AB - BACKGROUND: We sought to identify the relationship between response to previous systemic treatment and the efficacy of subsequent pemetrexed therapy in advanced non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: Two hundred and fifty clinical stage IIIB or IV NSCLC patients treated with pemetrexed as a second-line or further-line treatment between April 2007 and June 2008 were analyzed retrospectively. Prior therapies were divided into four types (gemcitabine-based [G], paclitaxel-based [P], docetaxel-based [D], and EGFR tyrosine kinase inhibitor [I]). Objective response rates (ORR) and progression-free survivals (PFS) for pemetrexed therapy were analyzed according to the response outcome with each previous treatment. RESULTS: The ORR of pemetrexed therapy was higher for patients who had achieved partial response with previous [G] therapy than others (15.0% vs. 4.3%, p=0.02). In addition, median PFS for pemetrexed therapy was greater for responders to [G] than for nonresponders (3.0 months vs. 1.7 months, p=0.004). The longer PFS for responders to [G] was also shown in the analysis among patients with squamous cell carcinoma (3.2 months vs. 1.7 months, p=0.056). By univariate analyses, the variables of the responder to [G] therapy, female, adenocarcinoma, never smoking status, and ECOG performance status of 0-1 were good predictive factors for pemetrexed therapy in terms of PFS. Multivariate analysis revealed that only response to [G] had statistical significance (hazard ratio=0.62, p=0.006). CONCLUSION: Response outcome to prior [G] therapy might predict the efficacy of subsequent pemetrexed therapy in advanced NSCLC. PMID- 19674812 TI - Variable-density flow in heterogeneous porous media--laboratory experiments and numerical simulations. AB - Konz, M., Ackerer, P., Younes, A., Huggenberger, P., Zechner, E., 2009a. 2D Stable Layered Laboratory-scale Experiments for Testing Density-coupled Flow Models. Water Resources Research, 45. doi:10.1029/2008WR007118., a series of laboratory-scale 2D tank experiments were conducted and accurately simulated for density driven flow problems on homogeneous porous media. In the present work, we extended the numerical and experimental studies to heterogeneous problems. The heterogeneous porous medium was constructed with a low permeability zone in the centre of the tank and had well-defined parameters and boundary conditions. Concentration distributions were measured in high resolution using a photometric method and an image analysis technique. The numerical model used for the simulations was based on efficient advanced approximations for both spatial and temporal discretizations. The Method Of Lines (MOL) was used to allow higher order temporal discretization. Three different boundary conditions, corresponding to different localizations of the inflow and the outflow openings at the opposite edges of the tank, were applied to investigate different flow scenarios in the heterogeneous porous medium flow tank. Simulation results of all three density coupled experiments revealed a density-dependent behavior of dispersion. Thus, a reduction of dispersivites was required to obtain a good matching of the experimental data. The high quality of the experiments enabled a detailed testing of numerical variable-density flow codes under heterogeneous conditions. Therefore, the experiments were considered to be reliable benchmark tests. PMID- 19674813 TI - A detailed field-based evaluation of naphthenic acid mobility in groundwater. AB - An anaerobic plume of process-affected groundwater was characterized in a shallow sand aquifer adjacent to an oil sands tailings impoundment. Based on biological oxygen demand measurements, the reductive capacity of the plume is considered minimal. Major dissolved components associated with the plume include HCO(3), Na, Cl, SO(4), and naphthenic acids (NAs). Quantitative and qualitative NA analyses were performed on groundwater samples to investigate NA fate and transport in the subsurface. Despite subsurface residence times exceeding 20 years, significant attenuation of NAs by biodegradation was not observed based on screening techniques developed at the time of the investigation. Relative to conservative tracers (i.e., Cl), overall NA attenuation in the subsurface is limited, which is consistent with batch sorption and microcosm studies performed by other authors. Insignificant biological oxygen demand and low concentrations of dissolved As (<10 microg L(-1)) in the plume suggest that the potential for secondary trace metal release, specifically As, via reductive dissolution reactions driven by ingress of process-affected water is minimal. It is also possible that readily leachable As is not present in significant quantities within the sediments of the study area. Thus, for similar plumes of process-affected groundwater in shallow sand aquifers which may occur as oil sands mining expands, a reasonable expectation is for NA persistence, but minimal trace metal mobilization. PMID- 19674814 TI - Effect of cadmium on diaphorase activity and nitric oxide production in barley root tips. AB - The effect of Cd on NADPH-diaphorase activity and nitric oxide (NO) production was investigated in barley root tips. The Cd-induced increase of NADPH-diaphorase activity occurred at the elongation zone and increased further in the differentiation zone of barley root tips. This activity was associated primarily with the microsomal membrane fraction of crude extract. In situ analysis revealed that the diaphorase activity was localized in the metaxylem and metaphloem elements and to some cells of the pericycle and parenchyma of root tips. Cd induced NO generation was observed in pericycle, parenchymatic stelar cells and companion cells of protophloem. The results suggest that the Cd-induced generation of NO functions in Cd toxicity through the ectopic and accelerated differentiation of root tips, causing the shortening of the root elongation zone and a subsequent reduction in root growth. PMID- 19674815 TI - Liver X receptor activation restores memory in aged AD mice without reducing amyloid. AB - Alterations in cerebral cholesterol metabolism are thought to play a role in the progression of Alzheimer's disease (AD). Liver X receptors (LXRs) are key regulators of cholesterol metabolism. The synthetic LXR activator, T0901317 has been reported to improve memory functions in animal models for AD and to reduce amyloid-beta (Abeta) deposition in the brain. Here we provide evidence that long term administration of T0901317 to aged, 21-month-old APPSLxPS1mut mice restores impaired memory. Cerebral cholesterol turnover was enhanced as indicated by the increased levels of brain cholesterol precursors and the upregulation of LXR target genes Abca1, Abcg1, and Apoe. Unexpectedly, the improved memory functions in the APPSLxPS1mut mice after T0901317 treatment were not accompanied by a decrease in Abeta plaque load in the cortex or hippocampus DG, CA1 or CA3. T0901317 administration also enhanced cerebral cholesterol turnover in aged C57BL/6NCrl mice, but did not further improve their memory functions. In conclusion, long-term activation of the LXR-pathway restored memory functions in aged APPSLxPS1mut mice with advanced Abeta deposition. However the beneficial effects of T0901317 on memory in the APPSLxPS1mut mice were independent of the Abeta plaque load in the hippocampus, but were associated with enhanced brain cholesterol turnover. PMID- 19674816 TI - Discovery of potential ZAP-70 kinase inhibitors: pharmacophore design, database screening and docking studies. AB - The best ZAP-70 inhibitor model consists of four-pharmacophore features, (1) one hydrogen bond acceptor, (2) one hydrogen bond donor (3) one hydrophobic aliphatic and (4) one hydrophobic aromatic features. This model was validated against 110 known ZAP-70 inhibitors with a correlation of 0.902 as well as enrichment factor of 1.61 against a maximum value of 2. This model picked 4094 hits from a database of 238,819 molecules while 358 molecules were indicated as highly active. Subsequently, docking studies were performed on the hits and novel series of potent leads were suggested based on the interactions energy between ZAP-70 and the putative inhibitors which validated not only the virtual screening potential of the model but also identified the possible new Chemotypes. PMID- 19674817 TI - Synthesis, anticonvulsant and CNS depressant activity of some new bioactive 1-(4 substituted-phenyl)-3-(4-oxo-2-phenyl/ethyl-4H-quinazolin-3-yl)-urea. AB - Several new 1-(4-substituted-phenyl)-3-(4-oxo-2-phenyl/ethyl-4H-quinazolin-3-yl) urea were synthesized and screened for anticonvulsant, CNS depressant and sedative-hypnotic activity in the mice. After i.p. injection to mice at doses of 30, 100, and 300 mg/kg body weight synthesized compounds were examined in the maximal electroshock induced seizures (MES) and subcutaneous pentylenetetrazole (scPTZ) induced seizure models in mice. Spectroscopic data and elemental analysis were consistent with the newly synthesized compounds. The neurotoxicity was assessed using the rotorod method. Compounds E1, E6, E9, E12, P3, P4 and P6 were found to be active in the MES screen whereas E1, P4, P6 and P11 were found to be active in the scPTZ screen. All except E6, E11 and P6 showed more than 50% decrease in locomotor activity at 1h of compound administration via actophotometer screen. CNS depressant activity screened with the help of the forced swim method resulted into some potent compounds. All the compounds were found to exhibit potent CNS depressants activity as indicated by increased immobility time. It can be concluded that newly synthesized compounds possessed promising CNS activities. PMID- 19674818 TI - [Dyspnea secondary to pulmonary hypertension in a patient with splenic myeloid metaplasia]. AB - There are several possible pathophysiological links between the development of pulmonary hypertension and myelofibrosis with myeloid metaplasia. We report a woman with myelofibrosis and myeloid metaplasia who presented with dyspnea and massive, painful splenomegaly. Right heart catheterization evidenced pulmonary hypertension. Her management consisted of splenic irradiation associated to sildenafil. Dyspnea in patients with myelofibrosis and myeloid metaplasia can be secondary to pulmonary hypertension and conversely the differential diagnosis of pulmonary hypertension should include a myeloproliferative syndrome. PMID- 19674819 TI - Serum albumin and total lymphocyte count as predictors of outcome in hip fractures. AB - BACKGROUND & AIMS: Hip fractures are a significant cause of mortality and morbidity in the elderly. Malnutrition is a significant contributor to this, however no consensus exists as to the detection or management of this condition. We hypothesise that results of admission serum albumin and total lymphocyte count (TLC), as markers of Protein Energy Malnutrition (PEM) can help predict clinical outcome in hip fracture patients aged over 60 years. METHODS: This retrospective study evaluated the nutritional status of patients with hip fractures using albumin and TLC assays and analysed their prognostic relevance. Clinical outcome parameters studied were delay to operation, duration of in-patient stay, re admission and in-patient, 3- and 12-month mortality. RESULTS: Four hundred and fifteen hip fracture patients were evaluated. Survival data were available for 377 patients at 12 months. In-hospital mortality for PEM patients was 9.8%, compared with 0% for patients without. Patients with PEM had a higher 12-month mortality compared to patients who had normal values of both laboratory parameters (Odds Ratio 4.6; 95% CI: 1.0-21.3). Serum albumin (Hazard Ratio 0.932, 95% CI: 0.9-1.0) and age (Hazard Ratio 1.04, 95% CI: 1.0-1.1) were found to be significant independent prognostic factors of mortality by Cox regression analysis. CONCLUSIONS: These results highlight the relevance of assessing the nutritional status of patients with hip fractures at the time of admission and emphasises the correlation between PEM and outcome in these patients. PMID- 19674820 TI - A non-disease in need of a name. PMID- 19674821 TI - Accumulation of anticoagulant rodenticides in a non-target insectivore, the European hedgehog (Erinaceus europaeus). AB - Studies on exposure of non-targets to anticoagulant rodenticides have largely focussed on predatory birds and mammals; insectivores have rarely been studied. We investigated the exposure of 120 European hedgehogs (Erinaceus europaeus) from throughout Britain to first- and second-generation anticoagulant rodenticides (FGARs and SGARs) using high performance liquid chromatography coupled with fluorescence detection (HPLC) and liquid-chromatography mass spectrometry (LCMS). The proportion of hedgehogs with liver SGAR concentrations detected by HPLC was 3 13% per compound, 23% overall. LCMS identified much higher prevalence for difenacoum and bromadiolone, mainly because of greater ability to detect low level contamination. The overall proportion of hedgehogs with LCMS-detected residues was 57.5% (SGARs alone) and 66.7% (FGARs and SGARs combined); 27 (22.5%) hedgehogs contained >1 rodenticide. Exposure of insectivores and predators to anticoagulant rodenticides appears to be similar. The greater sensitivity of LCMS suggests that hitherto exposure of non-targets is likely to have been under estimated using HPLC techniques. PMID- 19674822 TI - Leaf size and surface characteristics of Betula papyrifera exposed to elevated CO2 and O3. AB - Betula papyrifera trees were exposed to elevated concentrations of CO(2) (1.4 x ambient), O(3) (1.2 x ambient) or CO(2) + O(3) at the Aspen Free-air CO(2) Enrichment Experiment. The treatment effects on leaf surface characteristics were studied after nine years of tree exposure. CO(2) and O(3) increased epidermal cell size and reduced epidermal cell density but leaf size was not altered. Stomatal density remained unaffected, but stomatal index increased under elevated CO(2). Cuticular ridges and epicuticular wax crystallites were less evident under CO(2) and CO(2) + O(3). The increase in amorphous deposits, particularly under CO(2) + O(3,) was associated with the appearance of elongated plate crystallites in stomatal chambers. Increased proportions of alkyl esters resulted from increased esterification of fatty acids and alcohols under elevated CO(2) + O(3). The combination of elevated CO(2) and O(3) resulted in different responses than expected under exposure to CO(2) or O(3) alone. PMID- 19674823 TI - [Leech-borne infection on a TRAM flap: a case report]. AB - Leeches are used worldwide to treat venous congestion of flaps. Aeromonas hydrophila infections are recognized complications of leech use. We report a new case of delayed leech-borne infection in mammary reconstruction by a Transverse Rectus Abdominis Myocutaneous flap (TRAM), which caused the flap loss. The use of prophylactic antibiotics is a way to prevent A.hydrophila infection (third generation cephalosporin, ciprofloxacin). This antibioprophylaxy must be followed until wound closure of the venous congested tissue. PMID- 19674824 TI - Erosion and abrasion of tooth-colored restorative materials and human enamel. AB - OBJECTIVES: The aim of this study was to investigate the effects of erosion and toothbrush abrasion on different restorative materials and human enamel. METHODS: Human enamel and 5 kinds of tooth-colored restorative materials were used. The restorative materials included three composite resins (Filtek Silorane, Tetric EvoCeram, and Tetric EvoFlow), a polyacid-modified composite (Dyract Extra), and a conventional glass-ionomer cement (Ketac Fil Plus). For each type of the material, 40 specimens were prepared and embedded in ceramic moulds and divided into four groups (n=10): control group (C), erosion group (E), abrasion group (A), and erosion-abrasion group (EA). The specimens were subjected to six daily erosive attacks (groups E and EA; citric acid, pH 2.3, 1 min) and/or six abrasive attacks (groups A and EA; toothbrush abrasion, 100 strokes, 1 min), while the control specimens (group C) were maintained in artificial saliva. After 10-day treatment, the substance loss and surface changes were determined by surface profilometry and scanning electron microscopy. RESULTS: Human enamel presented higher substance loss when compared to restorative materials. Generally, combined erosion-abrasion (EA) caused the highest substance loss, followed by erosion, abrasion, and storage in artificial saliva. Composite resin presented highest durability under erosive and/or abrasive attacks. Enamel and restorative materials showed degradation in groups E and EA through SEM observation. CONCLUSIONS: Toothbrush abrasion has a synergistic effect with erosion on substance loss of human enamel, polyacid-modified composite and glass-ionomer cement. The acid- and abrasive-resistance of human enamel was lower compared to restorative materials. PMID- 19674826 TI - Effect of caregiver gender, age, and feedback prompts on chest compression rate and depth. AB - BACKGROUND: Quality of chest compressions (CC) is an important determinant of resuscitation outcome for cardiac arrest patients. PURPOSE: To characterize the quality of CC performed by hospital personnel, evaluate for predictors of CC performance, and determine the effects of audiovisual feedback on CC performance. METHODS: Seven hundred and fifty four individuals participated in a CPR quality improvement challenge at 30 US hospitals. Participants performed 2min of CC on a manikin with an accelerometer-based system for measuring both rate (CC/min) and depth (in.) of CC (AED Plus:ZOLL Medical). Real-time audiovisual feedback was disabled. A subset of participants performed a second trial of CC with the audiovisual feedback prompts activated. RESULTS: Mean depth of CC was below AHA minimum guidelines (<1.5in.) for 34% (1.30+/-0.14in.) and above maximum guidelines (>2.0in.) for 12% of participants (2.20+/-0.22in.). Depth of CC was greater for male vs. female (p<0.001) and younger vs. older (p=0.009) but did not differ between ACLS, BCLS, and non-certified participants (p=0.6). Predictors of CC depth included CC rate (r(part)=-0.34, p<0.0001), gender (r(part)=0.13, p=0.001), and age (r(part)=-0.09, p=0.02). Mean depth of CC increased, mean rate decreased, and variance in CC depth and rate declined when feedback was used (p< or =0.0001 vs. without feedback). The percentage of CC performed within AHA guidelines (1.5-2in.) improved from 15 to 78% with feedback. CONCLUSIONS: The quality of CC performed by personnel at US hospitals as judged by their performance on a manikin is often suboptimal. Quality of CC can be improved with use of CPR feedback technologies. PMID- 19674825 TI - Effects of large volume, ice-cold intravenous fluid infusion on respiratory function in cardiac arrest survivors. AB - International guidelines for cardiopulmonary resuscitation recommend mild hypothermia (32-34 degrees C) for 12-24h in comatose survivors of cardiac arrest. To induce therapeutic hypothermia a variety of external and intravascular cooling devices are available. A cheap and effective method for inducing hypothermia is the infusion of large volume, ice-cold intravenous fluid. There are concerns regarding the effects of rapid infusion of large volumes of fluid on respiratory function in cardiac arrest survivors. We have retrospectively studied the effects of high volume cold fluid infusion on respiratory function in 52 resuscitated cardiac arrest patients. The target temperature of 32-34 degrees C was achieved after 4.1+/-0.5h (cooling rate 0.48 degrees C/h). During this period 3427+/-210 mL ice-cold fluid was infused. Despite significantly reduced LV-function (EF 35.8+/-2.2%) the respiratory status of these patients did not deteriorate significantly. On intensive care unit admission the mean PaO(2) was 231.4+/-20.6 mmHg at a F(i)O(2) of 0.82+/-0.03 (PaO(2)/F(i)O(2)=290.0+/-24.1) and a PEEP level of 7.14+/-0.31 mbar. Until reaching the target temperature of or = 10 copies of the gene encoding beta actin were measured in the urine sediment genomic DNA. Sensitivity, specificity, and predictive values of the MSP and cytology tests were assessed and compared. RESULTS AND LIMITATIONS: MSP assays performed on 466 of the 496 (94%) valid urine samples identified two genes, TWIST1 and NID2, that were frequently methylated in urine samples collected from BCa patients, including those with early-stage and low-grade disease. The sensitivity of this two-gene panel (90%) was significantly better than that of cytology (48%), with comparable specificity (93% and 96%, respectively). The positive predictive value and negative predictive value of the two-gene panel was 86% and 95%, respectively. CONCLUSIONS: Detection of the methylated TWIST1 and NID2 genes in urine sediments using MSP provides a highly (> or = 90%) sensitive and specific, noninvasive approach for detecting primary BCa. TRIAL REGISTRATION: BlCa-001 study - EudraCt 2006-003303-40. PMID- 19674834 TI - Ultrasound-enhanced drug dispersion through solid tumours and its possible role in aiding ultrasound-targeted cancer chemotherapy. AB - It has been demonstrated that inadequate dispersion of cancer chemotherapeutic drugs throughout the tissues of larger, relatively poorly vascularised tumours compromises the therapeutic effectiveness of such drugs. Recently we demonstrated that electric fields could be exploited to achieve dispersion of a cancer chemotherapeutic drug through relatively impermeable tissues of a poorly vascularised solid tumour model. Using a modified Sonidel SP100 sonoporator we demonstrate that ultrasound may enhance the toxicity of a cancer chemotherapeutic drug by dispersing the drug through relatively impermeable tissues of a non vascularised tumour model in vivo. We suggest that such a phenomenon may play a significant role in ultrasound targeting of cancer chemotherapeutic drugs, particularly in the treatment of solid tumours. PMID- 19674836 TI - In-situ degradation of sulphur mustard using (1R)-(-)-(camphorylsulphonyl) oxaziridine impregnated adsorbents. AB - Bis-2-chloroethyl sulphide (sulphur mustard or HD) is an extremely toxic and persistent chemical warfare agent. For in-situ degradation of HD and its analogues (simulants), i.e., dibutyl sulphide (DBS) and ethyl 2-hydroxyethyl sulphide (HEES), different adsorbents systems loaded with (1R)-(-) (camphorylsulphonyl) oxaziridine were prepared. Solution of sulphur mustard and its simulants was prepared in carbon tetrachloride and taken for uniform adsorption on the impregnated systems using incipient volume. Degradation kinetics monitored by GC/FID were found to be first-order. The half-life of degradation reactions for simulants was obtained in less than 30 and for HD in 120 min. From the studied kinetics it was observed that reaction was very rapid with simulants and decreased rate was found for HD. The order of reactivity of MgO/Oxa system for HD and simulants was found to be DBS>HEES>HD. Reaction products of the oxidation reaction of simulants and HD on adsorbents were extracted in dichloromethane and analysed by GC-MS. The products were found to be non-toxic sulphoxide. The objective of the study is to develop a reactive adsorbent for in-situ degradation of sulphur mustard which could be used in nuclear biological and chemical (NBC) filtration systems. PMID- 19674835 TI - Removal of lead (II) from aqueous environment by a fibrous ion exchanger: polycinnamamide thorium (IV) phosphate. AB - The objective of the present research was to synthesize, characterize and to investigate the removal efficiency of lead (II) ion from synthetic lead solution by a hybrid fibrous ion exchanger. In the present study polycinnamamide thorium (IV) phosphate was synthesized by co-precipitation method and was characterized using SEM, XRD, FTIR and TGA-DSC. To know the practical applicability, a detailed removal study of lead ion was carried out using the material. The removal of lead was 81.2% under optimum conditions. Adsorption kinetic study revealed that the adsorption process followed first order kinetics. Adsorption data were fitted to linearly transformed Langmuir isotherm with R(2) (correlation coefficient) >0.99. Thermodynamic parameters were also calculated to study the effect of temperature on the removal process. In order to understand the adsorption type, equilibrium data were tested with Dubinin-Radushkevich isotherm. PMID- 19674837 TI - Preparation of TiO(2)/Ag colloids with ultraviolet resistance and antibacterial property using short chain polyethylene glycol. AB - TiO(2)/Ag nano-antibacterial material was prepared at low temperature using polyethylene glycol (PEG-600) as reducing and stabilizing agent. The size and shape as well as the optical properties of the nano-materials were characterized with transmission electron microscopy (TEM) and UV-vis spectroscopy (UV-vis). The results showed that the average particle size of TiO(2) among these nano materials was around 50-150 nm, and the average particle size of nano-silver was around 20 nm. Formation of Ag nano-particles on the surface of TiO(2) was confirmed by X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS), and the antibacterial activity was also investigated. By the antibacterial activity study and ultraviolet resistance test, it is noted that growth inhibition rates against E. coli was 99.99% as the concentration of nano particles dispersion solution was 10 ppm, the minimum UV protective effect could be achieved as the concentration was 290 ppm. PMID- 19674838 TI - Degradation of C.I. Acid Orange 7 by ultrasound enhanced heterogeneous Fenton like process. AB - The effect of ultrasonic power density, goethite addition, hydrogen peroxide concentration, initial pH, hydroxyl radical scavenger, and initial dye concentration on the decolorization of C.I. Acid Orange 7 by ultrasound/goethite/H(2)O(2) process was investigated. The results showed that the decolorization rate increased with power density, goethite addition, and hydrogen peroxide concentration, but decreased with the increase of initial dye concentration. The ultrasonic power density, goethite addition, and initial dye concentration have little effect on decolorization efficiency after 30 min reaction, while the increase of hydrogen peroxide concentration results in the increase of decolorization efficiency. There existed an optimal initial pH to achieve the highest decololrization rate and decolorization efficiency. The presence of hydroxyl radical scavenger would inhibit the decolorization reaction. Only less than half of total organic carbon (TOC) was removed after 90 min reaction, indicating more aggressive conditions are required to achieve the complete mineralization than those employed to simply break the chromophore group. PMID- 19674839 TI - Sonochemical destruction of nonylphenol: effects of pH and hydroxyl radical scavengers. AB - Nonylphenols are water-stable endocrine disrupting compounds that inhibit the growth of sewage bacteria in biological processes. The study describes the decomposition of 4-n-nonylphenol (NP) in water by 20 kHz ultrasound with emphasis on the impacts of pH, concentration and OH scavengers. It was found that the rate of degradation was accelerated by alkalinization, but more so by the addition of hydroxide alkalinity than carbonate. The addition of low doses of CO(3)(2-) and t butyl alcohol as strong scavengers of OH was also found to accelerate the decomposition of NP. The observation was attributed to the generation of reactive CO(3)(2-) and CH(3) via pyrolysis of the additives in the cavity bubbles. The results also revealed that NP did not compete with OH scavenging agents when their relative concentration was low. In case of high frequency sonication (861 kHz) the competition was slightly effective (slowed down degradation) at an identical dose of t-butyl alcohol. The difference was attributed to shorter bubble life time at high frequencies leading to less violent/less energetic bubble collapse and lower yield of CH(3). PMID- 19674840 TI - Acute sensitivity of activated sludge bacteria to erythromycin. AB - The presence of antibiotics in water resources has been disturbing news for the stakeholders who are responsible for public health and the drinking water supply. In many cases, biological wastewater treatment plants are the final opportunity in the water cycle to trap these substances. The sensitivity of activated sludge bacteria to erythromycin, a macrolide widely used in human medicine was investigated in batch toxicity tests using a concentration range of 1-300 mg L( 1). Erythromycin, a protein synthesis inhibitor, has been found to significantly inhibit ammonification, nitritation and nitratation at concentrations higher than 20 mg L(-1). The degree of inhibition increased with greater concentrations of the antibiotic. Exposure to erythromycin also clearly affected heterotrophs, particularly filamentous bacteria, causing floc disintegration and breakage of filaments. Cell lysis was observed with the concomitant release of organic nitrogen (intracellular proteins) and soluble COD. Although erythromycin exhibits properties of a surfactant, this characteristic alone cannot explain the damage to heterotrophs: the effects from erythromycin were greater than those of Tween 80, a commonly used surfactant. Floc disruption can lead to the release of isolated bacteria, and possibly antibiotic resistance genes, into the environment. PMID- 19674841 TI - Thrombin receptor: An endogenous inhibitor of inflammatory pain, activating opioid pathways. AB - Serine proteases such as thrombin, trypsin and mast cell tryptase can act on different cell types through protease-activated receptors (PARs). These receptors have been shown to be implicated in several phenomena such as inflammation, platelet activation, immune response and atherosclerosis. Several studies recently reported PARs expression on neurons and some of them demonstrated that these receptors could interfere with nociception. The contribution of PAR(1) to inflammatory pain and the mechanism involved in this phenomenon were investigated. Intraplantar injection of PAR(1) agonist increased withdrawal latency and reduced response frequency to von Frey filaments, thus inhibiting nociceptive response to both mechanical and thermal stimuli in mice. PAR(1) agonist also reduced carrageenan-induced inflammatory hyperalgesia. The anti nociceptive effects of PAR(1) agonist were mediated by endogenous opioids, as this effect was inhibited by local injection of naloxone methiodide, and because intraplantar injection of PAR(1) agonist increased mRNA expression of the endogenous opioid precursor proenkephalin. However, PAR(1) agonist was not able to inhibit calcium signals in isolated sensory neurons exposed to pro-nociceptive agents. Finally, despite similar inflammatory parameters, PAR(1)-deficient mice showed a strong potentiation of inflammatory hyperalgesia induced by the intraplantar injection of either formalin or carrageenan, or in the chronic model of collagen-induced arthritis, compared to wild-type mice. This study highlights a previously unknown endogenous mechanism of analgesia, showing a central role for the thrombin receptor PAR(1) in the regulation of inflammatory pain and as an activator of opioid pathways. PMID- 19674843 TI - Indentation modulus and hardness of viscoelastic thin films by atomic force microscopy: A case study. AB - We propose a nanoindentation technique based on atomic force microscopy (AFM) that allows one to deduce both indentation modulus and hardness of viscoelastic materials from the force versus penetration depth dependence, obtained by recording the AFM cantilever deflection as a function of the sample vertical displacement when the tip is pressed against (loading phase) and then removed from (unloading phase) the surface of the sample. Reliable quantitative measurements of both indentation modulus and hardness of the investigated sample are obtained by calibrating the technique through a set of different polymeric samples, used as reference materials, whose mechanical properties have been previously determined by standard indentation tests. By analyzing the dependence of the cantilever deflection versus time, the proposed technique allows one to evaluate and correct the effect of viscoelastic properties of the investigated materials, by adapting a post-experiment data processing procedure well established for standard depth sensing indentation tests. The technique is described in the case of the measurement of indentation modulus and hardness of a thin film of poly(3,4-ethylenedioxythiophene) doped with poly(4 styrenesulfonate), deposited by chronoamperometry on an indium tin oxide (ITO) substrate. PMID- 19674842 TI - An fMRI case report of photophobia: activation of the trigeminal nociceptive pathway. AB - Photophobia, or painful oversensitivity to light, occurs in a number of clinical conditions, which range from superficial eye irritation to meningitis. In this case study, a healthy subject with transient photophobia (induced by the overuse of contact lenses) was examined using functional magnetic resonance imaging (fMRI). While being scanned in a darkened environment, the subject was presented with intermittent 6-s blocks of bright light. The subject was scanned twice, once during his photophobic state and once after recovery. The subject reported that the visual stimuli produced pain (pain intensity=3/10 and unpleasantness=7/10) only during the photophobic state. During photophobia, specific activation patterns in the trigeminal system were seen at the level of the trigeminal ganglion, trigeminal nucleus caudalis, and ventroposteromedial thalamus. The anterior cingulate cortex, a brain structure associated with unpleasantness, was also active during photophobia. After recovery from photophobia, no significant activations were detected in these areas. This study may contribute to a better understanding of the pathways involved in photophobia in the human condition. PMID- 19674844 TI - Host and environmental risk factors associated with Cryptosporidium scophthalmi (Apicomplexa) infection in cultured turbot, Psetta maxima (L.) (Pisces, Teleostei). AB - An epidemiological cohort study of Cryptosporidium scophthalmi in cultured turbot Psetta maxima L. of Northwestern Spain was conducted along a four-year period. Four different ongrowing cohorts were monitored monthly from introduction into the ongrowing tanks (10-50 g) until reaching market size (400-1400 g). The association of host and environmental factors with five categories of parasite abundance was assessed using a multivariable regression framework. Epidemiological factors assessed here were water temperature, weight, length, month of collection, season, age, origin, condition factor, water filtration, and status to the myxozoan Enteromyxum scophthalmi infection. E. scophthalmi was included into the analysis because it targets the same organ than C. scophthalmi and it was prevalent in the studied population. The multivariable analysis demonstrated the statistically significant association between several factors and parasite abundance. C. scophthalmi abundance was associated (P<0.05) with age, condition factor, season, and status to E. scophthalmi infection. Young animals, with poor condition factor, during spring or summer, and not infected with the myxozoan were most likely to be highly infected by C. scophthalmi. Inclusion of these four variables significantly (P<0.05) improved the model, compared to the model that did not include any of these epidemiological factors. Increasing levels of C. scophthalmi abundance were associated (P<0.01) with higher severity of C. scophthalmi-compatible lesions. The frequency of distribution of C. scophthalmi abundance was clearly right-skewed and fitted a negative binomial distribution, whereas the intensity of infection fitted a Poisson distribution. The quantification of the variance-to-mean ratio stratified by age demonstrated overdispersion for 8-16 months old fish, although this bivariate association is likely affected by several other factors, as suggested by the results of the multivariable analysis. The negative relation between C. scophthalmi abundance and status to E. scophthalmi infection suggests differences in the transmission, onset, and course of both infections. The coarse filtration used in some cohorts did not significantly affect the levels of infection. C. scophthalmi was probably introduced into the ongrowing tanks mainly with carrier fish, though the involvement of infective oocysts from the water supply cannot be disregarded. Infection prevalence and mean intensity decreased with fish age and a seasonal distribution was found. Results presented here will help to understand the epidemiology of C. scophthalmi in turbot, to estimate the expected levels of infection associated with presence or absence of epidemiological factors, and to quantify the impact that the disease may have on susceptible turbot populations. The multivariable model used here is more powerful than the visual inspection of graphics for exploring associations in cooperative processes and can be easily extended to the assessment of epidemiological associations in other population and parasitic diseases. PMID- 19674846 TI - S100B serum levels are closely correlated with body mass index: an important caveat in neuropsychiatric research. AB - Elevated blood levels of S100B in neuropsychiatric disorders have so far been mainly attributed to glial pathologies. However, increases or dysfunction of adipose tissue may be alternatively responsible. Our study assessed S100B serum levels in 60 adult subjects without a prior history of neuropsychiatric disorders. S100B concentrations were closely correlated with the body mass index (BMI, range 18-45 kg/m(2)) as well as levels of leptin and adipocyte-type fatty acid-binding protein (A-FABP/FABP4) that are well-known adipose-related factors. Effect sizes as measured by Cohen's d indicated medium (0.8 > d > 0.5) to strong effects (d > 0.9) of BMI on S100B blood levels. In conclusion, physiological S100B levels in humans appear to closely reflect adipose tissue mass, which should therefore be considered as an important confounding factor in clinical studies examining the role of S100B. PMID- 19674848 TI - Unexpected serious adverse reactions to the therapy with botulinum toxin might be due to hypersensitivity. PMID- 19674847 TI - Integrated network systems and evolutionary developmental endocrinology. AB - Endocrine system has been considered to be a linear one, but the 'real world endocrine system' is a complex system, which is difficult to investigate using conventional strategies, such as single nucleotide polymorphism, genome-wide analysis, or gene targeting in animals. Here we propose a new strategy to comprehend the endocrine system as a complex network system. We introduced several novel concepts, such as complex system, network analysis, systems biology and evolutionary medicine, into the comprehension of endocrine system as a whole complex network system. This system is considered to be a scale-free network with key molecules such as acetyl CoA, NAD or ATP as 'hubs'. This system is robust against simple mutations, but various complex diseases may attack hubs. The system is also 'fractals', since there exist similar network systems among cells, proteins, and transcription factors in the lower levels, and there are similar ones among disease and social network in the higher levels. We propose to call this model 'Integrated Network Systems and Evolutionary DEvelopmental ENdocrinology (INS-EDEN)'. This novel framework will facilitate us to develop a new approach for understanding and treatment of various complex diseases related to endocrinology, and identify a unified theory of complex diseases. PMID- 19674845 TI - High pregnancy anxiety during mid-gestation is associated with decreased gray matter density in 6-9-year-old children. AB - Because the brain undergoes dramatic changes during fetal development it is vulnerable to environmental insults. There is evidence that maternal stress and anxiety during pregnancy influences birth outcome but there are no studies that have evaluated the influence of stress during human pregnancy on brain morphology. In the current prospective longitudinal study we included 35 women for whom serial data on pregnancy anxiety was available at 19 (+/-0.83), 25 (+/ 0.9) and 31 (+/-0.9) weeks gestation. When the offspring from the target pregnancy were between 6 and 9 years of age, their neurodevelopmental stage was assessed by a structural MRI scan. With the application of voxel-based morphometry, we found regional reductions in gray matter density in association with pregnancy anxiety after controlling for total gray matter volume, age, gestational age at birth, handedness and postpartum perceived stress. Specifically, independent of postnatal stress, pregnancy anxiety at 19 weeks gestation was associated with gray matter volume reductions in the prefrontal cortex, the premotor cortex, the medial temporal lobe, the lateral temporal cortex, the postcentral gyrus as well as the cerebellum extending to the middle occipital gyrus and the fusiform gyrus. High pregnancy anxiety at 25 and 31 weeks gestation was not significantly associated with local reductions in gray matter volume.This is the first prospective study to show that a specific temporal pattern of pregnancy anxiety is related to specific changes in brain morphology. Altered gray matter volume in brain regions affected by prenatal maternal anxiety may render the developing individual more vulnerable to neurodevelopmental and psychiatric disorders as well as cognitive and intellectual impairment. PMID- 19674849 TI - Solid-pseudopapillary neoplasm of the pancreas with massive central calcification in an old man. AB - Solid-pseudopapillary neoplasm (SPN) is a rare pancreatic tumor primarily affecting women in their twenties. It is characterized by a well-demarcated or encapsulated mass, indolent behavior and favorable prognosis. Capsular or punctate calcification is occasionally observed. Reported herein is a case of SPN of the pancreas with massive calcification in a 76-year-old Japanese man. Macroscopically, the pancreatic tumor appeared to be a simple calcified nodule, but histological examination revealed that it was an epithelioid tumor with massive calcification. The tumor cells, forming nests and cords, had eosinophilic cytoplasm and small eccentric nuclei. They were immunohistochemically positive for vimentin, CD56 and neuron-specific enolase. Nuclear accumulation of beta catenin protein and a point mutation of the beta-catenin gene by genomic DNA sequencing confirmed that the tumor was SPN. This is a very rare case of pancreatic SPN with massive calcification in an old man. PMID- 19674850 TI - Expression profiling of integrins in lung cancer cells. AB - Integrins are heterodimeric transmembrane receptors consisting of 18 alpha and 8 beta subunits. Heterodimer composition of alpha and beta subunits has a potential for determining tumor subtypes of human lung cancer. The purpose of this study was to investigate the expression profile of integrins in lung cancer cells. Expression profiling of integrins in a panel of lung cancer cell line, including A549 (adenocarcinoma, ADC), Calu-1 (squamous carcinoma, SCC), H1650 (bronchioloalveolar carcinoma, BAC) and DMS-53 (small cell lung cancer, SCLC), was analyzed by cDNA microarrays, restriction analysis of gene expression (RAGE) and flow cytometry. Seventy-nine lung cancer specimens were used to further validate the data from cell lines using immunohistochemistry. Integrins are obviously expressed in a cell type-specific manner, such as alpha 3 in A549, Calu 1 and H1650 except in DMS53, alpha 4 in H1650, alpha 5 and beta1 in all cell lines. The integrins detected with cDNA microarrays were all unequivocally detected with RAGE and by flow cytometry at the protein level. In all lung cancer specimens, alpha 3 integrin was strongly expressed in ADC, SCC and BAC, but was infrequent in SCLC. alpha 4 integrin was solely expressed in BAC. alpha 5 and beta1 integrins were expressed in all four histological types of lung cancer specimens. Integrin alpha 3 and alpha 4 may be useful as diagnostic markers for adenocarcinoma, squamous cell carcinoma and bronchioloalveolar carcinoma. RAGE is a promising technique for studying the expression profiles of genes, such as integrins in cancer cells. PMID- 19674851 TI - Mucinous adenocarcinoma as heterologous element in intermediately differentiated Sertoli-Leydig cell tumor of the ovary. AB - Sertoli-Leydig cell tumor (SLCT) is a rare tumor involving the ovary. Approximately 20% of SLCT are associated with heterologous elements that are either of endodermal or mesodermal origin. The gastrointestinal-type epithelium is the most commonly described endodermal heterologous element. SLCT with benign and borderline mucinous neoplasm has been reported in the literature. However, SLCT with mucinous adenocarcinoma as heterologous element has been rarely documented. Herein, we describe a rare case of intermediately differentiated Sertoli-Leydig cell tumor with mucinous adenocarcinoma as the heterologous element in a 21-year-old woman. She presented with throbbing lower abdominal pain and was found to have a large, complex left ovarian mass on imaging studies. She underwent left salpingo-oophorectomy, appendectomy and lymph node staging. Gross examination of the surgical specimen showed a large, encapsulated, solid-cystic mass completely replacing the ovary. Microscopically, the tumor was composed of intermediately differentiated Sertoli-Leydig cell tumor and well-differentiated mucinous adenocarcinoma. Interestingly, the bulk of the tumor (more than 90%) was composed of mucinous adenocarcinoma, whereas the SLCT component comprised less than 10% of the total tumor. The mucinous adenocarcinoma expressed positivity for CK20, CEA, CDX2 and CK7, and the SLCT component was positive for inhibin expression. The histopathological features and results of immunostaining were consistent with the diagnosis of the intermediately differentiated SLCT with mucinous adenocarcinoma as the heterologous element. This case was a diagnostic challenge as more than 90% of the tumor was composed of mucinous adenocarcinoma and SLCT constituted only the minor part of the tumor. This feature was in contrast to the previously described two cases, where mucinous adenocarcinoma as heterologous element was present as microscopic foci. This case highlights the importance of identifying the SLCT component in a case of an apparently pure mucinous adenocarcinoma in a young patient. PMID- 19674852 TI - Substance use among non-fatally injured patients attended at emergency departments in Spain. AB - AIMS: To describe the prevalence of recent use of alcohol, medication, and illegal drugs among patients who attended emergency departments (EDs) as a result of suffering an injury due to any external mechanism and to identify factors associated with alcohol and drug use. METHODS: A cross-sectional study was conducted in eight university hospitals in Spain. Participants were adult patients admitted to a trauma ED. Oral fluid was used to test for psychoactive substances analyzed by gas chromatography-mass spectrometry. Socio-demographic data and information on circumstances of the injury were collected through interviews. RESULTS: The analysis included 1579 patients admitted to the EDs (56.4% men). Among young people (< 40 years), 21.4% of men and 8.5% of women were positive for any illegal substance, primarily cannabinoids or cocaine; 24.7% of men and 14.8% of women were positive for alcohol. Among patients > or = 40 years, 7.4% of men and 1.6% of women were positive for any illegal substance, and 16.3% and 11.0% respectively for alcohol. Prevalence of substance detected varied across mechanism of injury, gender and age group. Night-time injury was associated with substance use. CONCLUSIONS: A high proportion of injured patients who were treated in an EDs tested positive for psychoactive drugs. Routine testing at trauma departments would maximize the identification of patients who may benefit from referral to specialized addiction treatment centers, or brief interventions. PMID- 19674853 TI - Spontaneous myometrial contractility in cows suffering from endometritis Influence of localisation, smooth muscle layer and cycle phase. An in vitro study. AB - Contractility of the healthy bovine myometrium depends on the reproductive state. Furthermore, contractility is influenced by localisation and the direction of smooth muscle strips. However, little is known about the contractile behaviour of the uterus when affected by endometritis. In our study, myometrial specimens from the larger horn (near the corpus and near the tip) in cows suffering from endometritis in estrus (n=8) or diestrus (n=8) were collected after slaughter. Two strips were prepared from each region corresponding to the circular and the longitudinal muscle layers, respectively. The spontaneous contractility of these strips was recorded in an organ bath. To analyse the results, the 2.5h recordings were divided into five periods of 30 min each. The variables area under curve (AUC) and maximal (A(max)) and minimal amplitude (A(min)) were calculated separately for each period, and the results were analysed using a non-parametric model regarding the influence of cycle phase (estrus vs. diestrus), region (corpus vs. tip) and muscle layer (circular vs. longitudinal). The values of both AUC and A(max) increased significantly over time. Muscle layer had a significant effect on AUC (corpus, tip) and A(max) (tip): the values of circular layers were increased compared to longitudinal layers. Dividing the data into subgroups allowed us to analyse them additionally according to muscle layer: In longitudinal layers, A(max) was increased at the corpus as compared with the tip. In this model, the factor cycle phase did not produce any significant difference in spontaneous myometrial activity. However, data of all variables showed non significant higher values in estrus than in diestrus samples. PMID- 19674854 TI - Proton pump inhibitors, osteoporosis, and osteoporosis-related fractures. AB - Proton pump inhibitors (PPIs) are among the most commonly prescribed medications today with an excellent short-term safety profile. Recently, a number of studies from a variety of data sources have reported an association between PPI use and hip fractures. However, there is not yet any direct evidence of a causal link between PPI use and the development of hip fracture. In the following paper, we will review the recent studies which have described this association between PPI use and hip fracture, and discuss the evidence supporting the likelihood of this association being causal, using data from previous work on the effects of surgical and pharmacological inhibition of gastric acid secretion on calcium absorption and bone mineral density. We will conclude by summarizing the current state of evidence on the relationship between gastric acid inhibition and the risk of fracture, and suggest management strategies for patients who require the long-term use of gastric acid inhibiting medications who also may be at risk for metabolic bone disease and fracture. PMID- 19674855 TI - [Multiple imputation of missing at random data: General points and presentation of a Monte-Carlo method]. AB - BACKGROUND: Statistical analysis of a data set with missing data is a frequent problem to deal with in epidemiology. Methods are available to manage incomplete observations, avoiding biased estimates and improving their precision, compared to more traditional methods, such as the analysis of the sub-sample of complete observations. METHODS: One of these approaches is multiple imputation, which consists in imputing successively several values for each missing data item. Several completed data sets having the same distribution characteristics as the observed data (variability and correlations) are thus generated. Standard analyses are done separately on each completed dataset then combined to obtain a global result. In this paper, we discuss the various assumptions made on the origin of missing data (at random or not), and we present in a pragmatic way the process of multiple imputation. A recent method, Multiple Imputation by Chained Equations (MICE), based on a Monte-Carlo Markov Chain algorithm under missing at random data (MAR) hypothesis, is described. An illustrative example of the MICE method is detailed for the analysis of the relation between a dichotomous variable and two covariates presenting MAR data with no particular structure, through multivariate logistic regression. RESULTS: Compared with the original dataset without missing data, the results show a substantial improvement of the regression coefficient estimates with the MICE method, relatively to those obtained on the dataset with complete observations. CONCLUSION: This method does not require any direct assumption on joint distribution of the variables and it is presently implemented in standard statistical software (Splus, Stata). It can be used for multiple imputation of missing data of several variables with no particular structure. PMID- 19674856 TI - [Gastro-esophageal reflux through gastric antisecretory drugs]. AB - During the 20th century, gastro-esophageal reflux moved from the status of a rare and severe disease to that of a frequent disease occurring mostly, in the absence of any significant lesions. Proton pump inhibitors (PPIs) are the mainstay of its therapy and are prescribed mainly in an empirical way. Extradigestive manifestations require more accurate diagnostic tests and therapeutic management. The modalities of prescription of the PPIs quickly progressed toward the on demand therapy and over-the-counter PPIs should become widespread. The relative failures of PPIs led to a profusion of new antisecretory agents but clinical improvements are presently disappointing and the rationale of this escalation is questionable. The concept of non acid gastro-esophageal reflux opens more innovative diagnostic and therapeutic perspectives which, however, must be validated. In this respect, endoscopic treatment needs more reliable techniques and more rigorous trials. Gastro-esophageal reflux refractory to PPIs corresponds mainly to functional esophageal disorders that need diagnostic and therapeutic improvements. Barrett's esophagus constitutes a major challenge for the next few years. Its screening and its prevention seem, for the moment, inaccessible. Its survey and the prevention of its complications should benefit from progress of diagnostic and interventional endoscopy. PMID- 19674857 TI - Overestimation of contralateral internal carotid artery stenosis before ipsilateral surgical endarterectomy. AB - OBJECTIVE: The aim of this study was to investigate a possible overestimation of the degree of contralateral carotid artery stenosis by duplex in patients with significant bilateral carotid stenoses who are to undergo carotid endarterectomy (CEA). METHODS: A retrospective analysis was performed of all patients undergoing CEA in our center over a period of 11 years. Pre- and postoperative duplex ultrasonography measurements of peak systolic velocity (PSV) and end diastolic velocity (EDV) were compared and used to classify the degree of stenosis. Univariate analysis was performed to indicate possible predictors for contralateral stenosis overestimation. RESULTS: A total of 384 CEA procedures in 357 patients were performed in our hospital. Pre- and postoperative bilateral duplex measurements were available in 135 patients. Forty-four out of 135 patients (33%) were preoperatively identified as having significant stenosis (>60%) of the internal carotid artery on both sides. In these patients, postoperative duplex measurements of the contralateral carotid showed a decrease in mean (SD) PSV from 2.53 (1.11) m s(-1) to 1.97 (0.87) m s(-1) (P<0.01) and a decrease in EDV from 0.87 (0.60) m s(-1) to 0.60 (0.36) m s(-1) (P<0.01). The absolute changes in contralateral PSV and EDV after CEA were larger among patients with a higher degree of stenosis preoperatively. These changes led to reclassification of stenosis to a lesser degree in 24 (55%) patients. In 16 cases (36%), this resulted in a measured stenosis on the contralateral side of less than 60%. CONCLUSIONS: One-third of the patients with duplex measurements consistent with bilateral significant carotid stenosis did not have a significant contralateral stenosis by duplex after CEA. Therefore, additional postoperative duplex measurement is advisable before planning contralateral CEA. PMID- 19674858 TI - Isolation of a new butanol-producing Clostridium strain: high level of hemicellulosic activity and structure of solventogenesis genes of a new Clostridium saccharobutylicum isolate. AB - New isolates of solventogenic bacteria exhibited high hemicellulolytic activity. They produced butanol and acetone with high selectivity for butanol (about 80% of butanol from the total solvent yield). Their 16S rDNA sequence was 99% identical to that of Clostridium saccharobutylicum. The genes responsible for the last steps of solventogenesis and encoding crotonase, butyryl-CoA dehydrogenase, electron-transport protein subunits A and B, 3-hydroxybutyryl-CoA dehydrogenase, alcohol dehydrogenase, CoA-transferase (subunits A and B), acetoacetate decarboxylase, and aldehyde dehydrogenase were identified in the new C. saccharobutylicum strain Ox29 and cloned into Escherichia coli. The genes for crotonase, butyryl-CoA dehydrogenase, electron-transport protein subunits A and B, and 3-hydroxybutyryl-CoA dehydrogenase composed the bcs-operon. A monocistronic operon containing the alcohol dehydrogenase gene was located downstream of the bcs-operon. Genes for aldehyde dehydrogenase, CoA-transferase (subunits A and B), and acetoacetate decarboxylase composed the sol-operon. The gene sequences and the gene order within the sol- and bcs-operons of C. saccharobutylicum Ox29 were most similar to those of Clostridium beijerinckii. The activity of some of the bcs-operon genes, expressed in heterologous E. coli, was determined. PMID- 19674859 TI - Development of a liquid chromatographic system with fluorescent detection for beta-secretase immobilized enzyme reactor on-line enzymatic studies. AB - A novel liquid chromatographic method has been developed for use in throughput screening of new inhibitors of human recombinant beta-amyloid precursor protein cleaving enzyme (hrBACE1). The approach is based on the use of an immobilized enzyme reactor (IMER) containing the target enzyme (hrBACE1-IMER) and uses fluorescence detection. The bioreactor was prepared by immobilizing hrBACE1 on an ethylendiamine (EDA) monolithic disk (CIM) and a fluorogenic peptide (M-2420) containing the beta-secretase site of the Swedish mutation of amyloid precursor protein (APP) was used as substrate. After injection into the hrBACE1-IMER system, M-2420 was enzymatically cleaved, giving rise to a fluorescent methoxycoumaryl-fragment (Rt=1.6min), which was separated from the substrate and selectively detected at lambda(exc)=320 and lambda(em)=420nm. Product and substrate were characterized by using a post monolithic C18 stationary phase coupled to an ion trap mass analyser. A calibration curve was constructed to determine the immobilized hrBACE1-IMER rate of catalysis and kinetic constants. Specificity of the enzymatic cleavage was confirmed by injecting the substrate on a blank CIM-EDA. The proposed method was validated by the determination of the inhibitory potency of five reference compounds with activities ranked over four order of magnitude (four peptidic inhibitors and a green tea polyphenol, ( )gallocatechin gallate). The obtained results were found in agreement with the data reported in literature, confirming the validity and the applicability of the hrBACE1-IMER as a tool for the fast screening of unknown inhibitors (more than 6 compounds per hour). Moreover, the hrBACE1-IMER showed high stability during the analysis, permitting its use for more than three months without affecting enzyme activity. PMID- 19674860 TI - Teaching medical interviewing to patients: the other side of the encounter. AB - OBJECTIVES: This paper describes the development and pilot testing of a communication skills curriculum based on medical student curriculum and modified for use with patients. METHODS: Six key concepts from our introductory medical education communication skills curriculum were identified. The core knowledge and skills related to these concepts were reorganized into six modules, including presentation materials, handouts and active learning components. The curriculum was pilot tested with three independent groups of non-medical participants, representing a broad cross-section of the community. RESULTS: Participants reported a high level of satisfaction; over 80% found the program helpful for learning new information and skills, and 92% for working with their physicians. Over 90% would recommend the program to others. Participant self-assessments revealed the greatest change in knowledge of medical interviewing. Skill changes were greatest in expressing emotion and efficiently telling the medical story. The pilot tests also highlighted the importance of other issues related to recruitment and health literacy. CONCLUSIONS: This project demonstrates that key concepts underlying doctor-patient communications can be simplified and repackaged for use from the patient's perspective. PRACTICE IMPLICATIONS: Similar curricula can empower patients from all walks of life to better communicate with their health care providers and enhance their healthcare experience. PMID- 19674861 TI - A pilot study assessing emotional intelligence training and communication skills with 3rd year medical students. AB - OBJECTIVE: To investigate whether emotional intelligence (EI) developmental training workshops can lead to increases with the Bar-On Emotional Quotient (EQ i) total scores. METHODS: A pilot study with a quasi-randomised controlled design was employed with self-report assessments conducted at baseline and post intervention following a 7-month training programme. Medical students based at a UK-based medical school participated in the study, and 36 volunteer students were recruited to the control group with 50 students randomly assigned to receive the intervention. A total of 34 (68%) students in the intervention group attended the first intervention training workshop, 17 (34%) attended the majority of the monthly development sessions and completed the post-intervention assessment. In the control group only one participant did not complete the follow-up assessment. RESULTS: The intervention group had significantly higher EQ-i change from baseline mean scores than the control group. The intervention group mean scores had increased across time, whilst the control group mean scores slightly decreased. CONCLUSION: The EI developmental training workshops had a positive effect on the medical students in the intervention group. PRACTICE IMPLICATIONS: Further research is warranted to determine whether EI can be a useful measure in medical training, and the concept and measurement of EI requires further development. PMID- 19674862 TI - Encouraging patients with depressive symptoms to seek care: a mixed methods approach to message development. AB - OBJECTIVE: To assess the message preferences of individuals affected by depression as part of a project that will evaluate interventions to encourage at risk patients to talk to their physicians about depression. METHODS: Adaptive Conjoint Analysis (ACA) of 32 messages defined by 10 message attributes. Messages were developed based on input from three focus groups comprised of individuals with a personal and/or family history of depression, then tested using volunteers from an Internet health community. In an online conjoint survey, 249 respondents with depression rated their liking of the messages constructed for each attribute. They were then presented with two message sets and rated their preferences. Preference utilities were generated using hierarchical Bayes estimation. RESULTS: The optimal communication approach described both psychological and physical symptoms of depression, recognized multiple treatment options, offered lifetime prevalence data, noted that depression can affect anyone, and acknowledged that finding an effective treatment can take time. CONCLUSION: Individuals with depression respond differently to depression care messages, underscoring the need for careful message development and evaluation. PRACTICE IMPLICATIONS: ACA, used in conjunction with focus groups, is a promising approach for developing and testing messages in the formative research stage of intervention development. PMID- 19674863 TI - Mind the gap: Learners' perspectives on what they learn in communication compared to how they and others behave in the real world. AB - OBJECTIVE: To explore the lack of the learners' voice at previous international conferences on communication in healthcare. METHODS: A group of medical students and recently qualified junior doctors were invited to give the learner's perspective on how communication skills are taught and how they are implemented in 'real life', at a 90min symposium at the EACH International Conference on Communication in Healthcare, 4th September 2008, Oslo. RESULTS: We attempt to bridge the gap between learning communication skills formally in the medical classroom and actually implementing these in the real world between doctors and patients from a learners' perspective. In making this transition we highlight obvious weaknesses and potential pitfalls, whilst also drawing attention to the successful strategies used in our respective medical schools. Four key areas are discussed: (1) using simulated patients, (2) learning in the clinical setting, (3) barriers to utilizing communication skills, (4) future directions. We have drawn upon the learning experiences from both undergraduate and postgraduate environments in the UK, the USA and Norway. CONCLUSION: Our experiences differed between universities from the same country, which widened across continents. The differences between how we behave in the classroom and how we are with real patients when unobserved have been highlighted; and we have attempted to explain why trainees sometimes modify their behavior in medical assessments with standardized patients for examinations as opposed to how we would perform on wards or in general practice. The teaching of communication skills will continue to develop over the forthcoming years. PRACTICE IMPLICATIONS: Integrating communication skills into medical school curricula is essential. Identifying enthusiastic doctors who are effective communicators and have the initiative to help develop this is vital. It may be beneficial to train simulated patients to react to students in a variety of different ways to reflect the diversity of true patient responses. In addition, having a better understanding of the multidisciplinary roles and rapidly developing technology would facilitate not only communication between health professionals but would also help optimize patient care. PMID- 19674864 TI - Improvement in insulin resistance and reduction in plasma inflammatory adipokines after weight loss in obese dogs. AB - Obesity is now a major disease of dogs, predisposing to numerous disorders including diabetes mellitus. Adipocytes are active endocrine cells, and human obesity is characterized by derangements in inflammatory adipokine production. However, it is unclear as to whether similar changes occur in dogs. The purpose of the current study was to assess insulin sensitivity and inflammatory adipokine profiles in dogs with naturally occurring obesity and to investigate the effect of subsequent weight loss. Twenty-six overweight dogs were studied, representing a range of breeds and both sexes. All dogs underwent a weight loss program involving diet and exercise. Body fat mass was measured by dual-energy x-ray absorptiometry; plasma concentrations of insulin, glucose, and a panel of inflammatory adipokines (including acute-phase proteins, cytokines, and chemokines) were also analyzed. Body fat mass before weight loss was positively correlated with both plasma insulin concentrations (Kendall tau=0.30, P=0.044) and insulin:glucose ratio (Kendall tau=0.36, P=0.022), and both decreased after weight loss (P=0.0037 and 0.0063, respectively). Weight loss also led to notable decreases in plasma tumor necrosis factor-alpha (TNF-alpha), haptoglobin, and C reactive protein concentrations (P<0.05 for all), suggesting improvement of a subclinical inflammatory state associated with obesity. This study has demonstrated that in obese dogs, insulin resistance correlates with degree of adiposity, and weight loss improves insulin sensitivity. Concurrent decreases in TNF-alpha and adipose tissue mass suggest that in dogs, as in humans, this adipokine may be implicated in the insulin resistance of obesity. PMID- 19674865 TI - Interactions between genes involved in growth and muscularity in pigs: IGF-2, myostatin, ryanodine receptor 1, and melanocortin-4 receptor. AB - In the swine breeding industry, two economical traits are of particular importance in sires, namely, muscle growth and average daily gain (ADG). These traits are quantitative, which implies that they are under the control of multiple genes. Mutations in these genes, associated with either muscularity or growth, are useful quantitative trait nucleotides (QTN) for unraveling genetic variation of these traits and can be used in marker-assisted selection. Until now, QTN involved in muscle growth and/or ADG in pigs were identified in porcine ryanodine receptor 1 (RYR1), insulin-like growth factor-2 (IGF-2), and melanocortin-4 receptor (MC4R). Recently, a fourth possible QTN was found in porcine myostatin (MSTN). All four QTN have an influence on muscle growth and/or somatic growth, so an influence of one mutation on one or more of the other mutations should not be excluded. However, although the polymorphisms in the RYR1 and the MC4R gene affect the function of the respective protein, the polymorphisms of the IGF-2 and MSTN gene influence the mRNA expression of the respective gene. Therefore, this study investigated possible interactions between the genotypes of MSTN, IGF-2, and MC4R (population 1) or the RYR1, IGF-2, and MSTN QTN (population 2) on IGF-2 and MSTN expression in different muscle types in pigs. In both skeletal muscle and heart muscle growth, the IGF-2:MSTN ratio seems to play an important role. Also, the RYR1 genotype had a significant effect on IGF-2 expression in m. longissimus dorsi. No effect of the MC4R QTN could be seen. PMID- 19674866 TI - Peptide screening of cerebrospinal fluid in patients with glioblastoma multiforme. AB - AIMS: To apply modern mass spectrometry based technology to identify possible CSF peptide markers of glioblastoma multiforme (GBM). METHODS: Mass spectrometry based peptidomics technology enables a systematic and comprehensive screening of cerebrospinal fluid (CSF) with regard to its peptide composition. Differential Peptide Display (DPD) allows the identification of single marker peptides for a target disease. Using both, we analyzed CSF samples of 11 patients harbouring a glioblastoma multiforme in comparison to 13 normal controls. RESULTS: Four CSF peptides which significantly distinguished GBM from controls in all applied statistic tests could be identified out of more than 2,000 detected CSF peptides. They were specific C-terminal fragments of alpha-1-antichymotrypsin, osteopontin, and transthyretin as well as a N-terminal residue of albumin. All molecules are constituents of normal CSF, but none has previously been reported to be significantly elevated in CSF of GBM patients. CONCLUSION: The study showed that peptidomics technology is able to identify possible biomarkers of neoplastic CNS disease. It remains to be determined if the identified elevated CSF peptides are specific for GBM. With regard to GBM, however, the more important role of CSF peptide biomarkers than aiding initial diagnosis might be early recognition of disease recurrence or monitoring of efficacy of adjuvant therapy protocols. PMID- 19674867 TI - [A new drug of abuse: Gammabutyrolactone (GBL)]. PMID- 19674868 TI - [Liver transplantation for hepatocellular carcinoma and for patients with pre existing cancers]. AB - Hepatocellular carcinoma (HCC) is currently the leading indication for liver transplantation in France, accounting for 25% of all cases. Transplantation is appropriate, nonetheless, only for patients whose HCC has a low risk of posttransplant recurrence and is limited in size and number, meeting the Milan criteria (1 single nodule of a maximum diameter of 5 cm or 3 lesions of a maximum diameter of 3 cm), or slightly exceeds these criteria without vascular invasion visible on preoperative imaging. Results for this indication are very satisfactory, and 5-year survival ranges from 60 to 80%, according to tumor stage. Small HCCs (<2 cm) are usually treated conservatively. Transplantation is proposed in cases of a contraindication to resection or radiofrequency ablation or of recurrence after local treatment. A history of an extrahepatic tumor is found in approximately 5% of candidates for liver transplantation. This history is not necessarily a contraindication to transplantation. They may be considered eligible for a graft after discussion in a multidisciplinary meeting, if the extrahepatic tumor was treated curatively and if their 5-year tumor-related life expectancy is greater than 50-60%. PMID- 19674869 TI - Attentional control theory: anxiety, emotion, and motor planning. AB - The present study investigated how trait anxiety alters the balance between attentional control systems to impact performance of a discrete preplanned goal directed motor task. Participants executed targeted force contractions (engaging the goal-directed attentional system) at the offset of emotional and non emotional distractors (engaging the stimulus-driven attentional system). High and low anxious participants completed the protocol at two target force levels (10% and 35% of maximum voluntary contraction). Reaction time (RT), performance accuracy, and rate of change of force were calculated. Expectations were confirmed at the 10% but not the 35% target force level: (1) high anxiety was associated with slower RTs, and (2) threat cues lead to faster RTs independently of trait anxiety. These new findings suggest that motor efficiency, but not motor effectiveness is compromised in high relative to low anxious individuals. We conclude that increased stimulus-driven attentional control interferes with movements that require greater attentional resources. PMID- 19674870 TI - Serum uric acid, hyperuricemia and body mass index in children and adolescents with intellectual disabilities. AB - The aims of the preset study were to describe the profile of serum uric acid, the prevalence of hyperuricemia and its risk factors among children and adolescents with intellectual disabilities. We conducted a cross-sectional study of 941 children and adolescents with intellectual disabilities (aged 4-18 years) who participated in annual health examinations in three special schools in Taiwan. This study indicated 30.6% boys and 17.9% girls with intellectual disabilities were with hyperuricemia in Taiwan. The factors of gender, age and BMI were variables that can significantly predict the hyperuricemia occurrence in this vulnerable population. Those children and adolescents with intellectual disabilities were boys (OR=2.93, 95% CI=2.02-4.26) and older age (OR=6.49, 95% CI=2.19-19.21) were more likely to be hyperuricemia. With regard to BMI to hyperuricemia occurrence, those children and adolescents with intellectual disabilities were overweight (OR=1.16-3.21, 95% CI=1.16-3.21) and being obese (OR=4.95-11.58, 95% CI=4.95-11.58) was more likely to have a hyperuricemia than the normal weight group. This study provides the general profile of serum uric acid, hyperuricemia and its risk factors of children and adolescents with intellectual disabilities. Medical professionals should be highly alert to the possible consequences of hyperuricemia and provide useful information about the clinical manifestation of this condition for caregivers of children and adolescents with intellectual disabilities. PMID- 19674871 TI - Precision-guided surgical navigation system using laser guidance and 3D autostereoscopic image overlay. AB - This paper describes a precision-guided surgical navigation system for minimally invasive surgery. The system combines a laser guidance technique with a three dimensional (3D) autostereoscopic image overlay technique. Images of surgical anatomic structures superimposed onto the patient are created by employing an animated imaging method called integral videography (IV), which can display geometrically accurate 3D autostereoscopic images and reproduce motion parallax without the need for special viewing or tracking devices. To improve the placement accuracy of surgical instruments, we integrated an image overlay system with a laser guidance system for alignment of the surgical instrument and better visualization of patient's internal structure. We fabricated a laser guidance device and mounted it on an IV image overlay device. Experimental evaluations showed that the system could guide a linear surgical instrument toward a target with an average error of 2.48 mm and standard deviation of 1.76 mm. Further improvement to the design of the laser guidance device and the patient-image registration procedure of the IV image overlay will make this system practical; its use would increase surgical accuracy and reduce invasiveness. PMID- 19674872 TI - A vectorial image soft segmentation method based on neighborhood weighted Gaussian mixture model. AB - The CT uroscan consists of three to four time-spaced acquisitions of the same patient. After registration of these acquisitions, the data forms a volume in which each voxel contains a vector of elements corresponding to the information of the CT uroscan acquisitions. In this paper we will present a segmentation tool in order to differentiate the anatomical structures within the vectorial volume. Because of the partial volume effect (PVE), soft segmentation is better suited because it allows regions or classes to overlap. Gaussian mixture model is often used in statistical classifier to realize soft segmentation by getting classes probability distributions. But this model relies only on the intensity distributions, which will lead a misclassification on the boundaries and on inhomogeneous regions with noise. In order to solve this problem, a neighborhood weighted Gaussian mixture model is proposed in this paper. Expectation maximization algorithm is used as optimization method. The experiments demonstrate that the proposed method can get a better classification result and is less affected by the noise. PMID- 19674873 TI - In vitro analysis of the occurrence of a paradoxical effect with different echinocandins and Candida albicans biofilms. PMID- 19674874 TI - One-pot biomimetic synthesis of monolithic titania through mineralization of polysaccharide. AB - To obviate the common precipitation of titania after a precursor addition into aqueous or water-containing solutions, here we are using ethylene glycol as the reaction media with dissolved polysaccharide xanthan. The water is introduced in a restricted amount to provide only the hydration of polysaccharide macromolecules. An introduced precursor is involved into instant hydrolysis and following condensation reactions where contacting with the hydrating water. This resulted in titania formation on carbohydrate macromolecules like the biomineralization in living organisms. The gelling of solution proceeded without the precipitation even when the precursor was taken in concentration of ca. 3 wt.%. Polysaccharide in these syntheses served as a template. By varying the concentrations of precursor, xanthan and water, it was possible to manipulate the metal oxide morphology. Fibrillar, particulate and plate-like structures presented that depended on the synthesizing conditions. Titania prepared at ambient conditions was amorphous. When it was calcined at temperatures between 300 and 900 degrees C, crystalline anatase and rutile were found at 300 and 700 degrees C, respectively. PMID- 19674875 TI - Fungal based synthesis of silver nanoparticles--an effect of temperature on the size of particles. AB - A simple and effective approach to aqueous based biosynthesis of silver nanoparticles was demonstrated and the effect of temperature on controlling size of silver nanoparticles was studied. The morphology and uniformity of silver nanoparticles were investigated by UV-Vis spectroscopy, X-ray diffraction and HrTEM. The functional group of protein molecule was identified using FTIR. Increase in reaction temperature leads to decrease in size of silver nanoparticles and increase in monodispersity. PMID- 19674876 TI - Phospholipidosis in healthy subjects participating in clinical studies: ultrastructural findings in white blood cells. AB - Lipid storage disorders and phospholipidosis share similar morphologic characteristics displayed as lamellar bodies at ultrastructural level. More than 50 cationic amphiphilic drugs (CADs), including antidepressants, antianginal, antimalarial, and cholesterol-lowering agents, have been reported to induce phospholipidosis, however, the mechanism by which this occurs has not been extensively studied and is not well understood. Both the Food and Drug Administration (FDA) and the pharmaceutical industry recognized drug-induced phospholipidosis as a significant challenge for drug development. In a randomized, double-blind, placebo-controlled, active-controlled, ascending multiple-dose study to investigate the tolerability, safety, pharmacokinetics, and pharmacodynamics of a new investigational drug (an antihypertensive drug in early drug development) in healthy male subjects, possible drug-induced phospholipidosis was also explored ultrastructurally. Given the presence of these structures both pretreatment and following placebo treatment, it was concluded that the presence of phospholipid-like structures in individual volunteers could be a normal background finding in neutrophilic granulocytes thus emphasizing their role as natural phagocytic cells. Recommendations for the conduct of this type of studies are given. PMID- 19674877 TI - Subchronic exposure to arsenic trioxide-induced oxidative DNA damage in kidney tissue of mice. AB - To evaluate the oxidative DNA damage in kidney tissue of mice exposed to arsenic trioxide (As) subchronically, expression of 8-hydroxy-2-deoxyguanosine (8-OHdG) and pathologic changes were observed. Forty mice were randomly divided into 4 groups of 10 each (5 mice of each sex). Group 1 received drinking water alone (control). Groups 2, 3 and 4 received 1, 2 and 4 mg/L arsenic trioxide, respectively. Arsenic trioxide was given through drinking water for 60 days. The expression of 8-OHdG in the kidney tissue of mice was analyzed observed in these 4 groups. The groups treated with As showed pathologic changes in the kidney tissue and significant increase in the level of 8-OHdG expression (P<0.01). Moreover, the dose-dependent increase between As exposure and renal damages were observed. Especially, its immunoactivity was strong in the proximal convoluted tubule and Bowman's capsule. These results suggest that chronic exposure to As induces damages to kidney tissue, and especially the epithelial cells of proximal convoluted tubule and the podocytes of the Bowman's capsule may be more sensitive to As-induced nephrotoxicity. PMID- 19674878 TI - Toxicological evaluation of 10% Solanum lycocarpum St. Hill fruit consumption in the diet of growing rats: hematological, biochemical and histopathological effects. AB - Solanum lycocarpum St. Hill (Solanaceae) is a native shrub very common in the Brazilian savanna. The fruit of this plant contains steroidal glycoalkaloids that may disrupt the endocrine system. Because this plant is employed in folk medicine for the management of diabetes, obesity and decreasing cholesterol levels, the present study determined the possible toxic effects of exposure to S. lycocarpum fruit from weaning (21 days old) until adult age (8 weeks of treatment) in male and female rats. In male rats, the plant reduced weight gain, while few significant differences were observed in female animals. Slight significant differences were observed in food and water consumption and in hematological parameters in treated rats. Reductions in adrenal gland, spleen, heart, kidneys and thymus weights of treated males were observed, while increased relative weights were detected in the heart, epididymises, lungs, seminal vesicles, and testicles. In females, no differences were observed in organ weights and few differences were observed in relative weights of some organs. The histopathologic study showed no alteration between groups. Serum biochemical parameters showed triglyceride reductions in treated animals of both sexes; in females, an increase in albumin and alanine aminotransferase levels and a reduction in total protein levels were noted. The present data therefore demonstrate sex-related differences in S. lycocarpum toxicity. PMID- 19674879 TI - Ontogeny of photophore pattern in the velvet belly lantern shark, Etmopterus spinax. AB - Bioluminescence is known to be of great ecological importance to a luminous organism but extremely few studies investigate the ontogeny of luminous capabilities. The photogenic pattern of the velvet belly lantern shark Etmopterus spinax was investigated over ontogeny (14.0-52.5 cm total length) to determine the scaling of the surface area and the photophore density of different luminous zones as well as the ecological consequences of ontogenetic variations in bioluminescence efficiency. According to the luminous zone considered, different scaling patterns were found for the surface areas while the photophore densities of all zones scale with negative allometry, even though photophore insertion occurs. No sexual differences in these relationships were found. Luminous zones can be placed in two morphologically different groups: the "coverage" and the "isolated" zones. While counter-illumination is certainly the function of the former, the latter are probably involved in intraspecific behaviours. Due to the discrepancy between luminous capabilities of these two luminous zone categories, there is an ontogenetic increase in the luminescence heterogeneity of the luminous pattern as it was shown by luminescence modelling and confirmed by direct observations of spontaneous luminescence in living sharks. This heterogeneity certainly represents a trade-off between an efficient ventral camouflage and a strong identification tool for intraspecific behaviours such as coordinate hunting, which would be particularly useful when E. spinax become fish eaters (>19 cm total length), and for sexual recognition in mature individuals. PMID- 19674881 TI - Effect of xanthone from Kielmeyera coriacea stems on serotonergic neurons of the median raphe nucleus. AB - Kielmeyera coriacea Mart. (Clusiaceae), known as "Pau Santo", is used to treat several tropical diseases. The hydroethanolic extract (HE) of Kielmeyera coriacea stems and its semi-pure dichloromethane constituent (DCM) produced an anti immobility effect in rats submitted to the forced swimming test (FST), suggesting a antidepressant-like profile. This study evaluated the effect of intra-median raphe nucleus (MRN) microinjection of 1,3,7-trihydroxy-2-(3-methylbut-2-enyl) xanthone, present in large quantity in the HE from Kielmeyera coriacea stems, on immobility behaviour in the FST in rats. The effects of xanthone were compared with intra-MRN microinjections of Way100635 (5-HT1A antagonist) or (+) 8-OH-DPAT (5-HT1A agonist). Locomotor activity in the open-field test (OFT) was evaluated as a complementary measure. Xanthone (0.3ng) or Way100635 (2.5microg) reduced, whereas (+) 8-OH-DPAT (5.0microg) increased immobility time in the FST. Way100635 (2.5 or 5.0microg) completely reversed the effects of (+) 8-OHDPAT (5.0microg), and potentiated the anti-immobility effect of the ineffective dose of xanthone (0.2ng) in the FST. The association of effective doses of (+) 8-OH-DPAT (5.0microg) and xanthone (0.3ng) annulled the effect of each compound on immobility time. These results suggest that xanthone acts as an antagonist at 5 HT1A autoreceptors in MRN and increases serotonin (5-HT) availability in projection regions, proving to be a prototype drug that may be useful in mood isorders such as depression, or indeed be a beneficial adjunctive treatment improving the efficacy and/or accelerating the effects of antidepressant drugs in patients with major depression. PMID- 19674882 TI - The role of EBUS-TBNA for the diagnosis of sarcoidosis--comparisons with other bronchoscopic diagnostic modalities. AB - BACKGROUND: The diagnosis of sarcoidosis requires both compatible clinical features and pathologic findings as a means to exclude other differential diagnoses. The utility of endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) for diagnosis of sarcoidosis has been reported, although its indication remains unclear for cases of suspicious sarcoidosis. To clarify the role of EBUS-TBNA for the diagnosis of sarcoidosis, we compared three diagnostic modalities: EBUS-TBNA, transbronchial lung biopsy (TBLB) and bronchoalveolar lavage fluid analysis (BAL). METHODS: Thirty-eight patients with suspicious sarcoidosis who had enlarged hilar and/or mediastinal lymph nodes on chest CT were retrospectively reviewed. Patients with malignancies or prior established diagnosis of sarcoidosis were excluded. BAL was initially performed followed by TBLB and finally EBUS-TBNA at the same setting. Microbacterial examinations were also performed from all samples. RESULTS: Pathological findings compatible with sarcoidosis were obtained in 32 patients. The remaining 6 patients were diagnosed as one case each of chronic eosinophilic pneumonia, atypical mycobacterial infection and tuberculosis, and the remaining three were pathologically indefinite cases. Clinically, 35 patients were diagnosed with sarcoidosis. The diagnostic accuracy of sarcoidosis was significantly better by EBUS-TBNA (91.4%, p<0.001) compared to the other two modalities. According to chest roentgenogram classifications, there were 31 stage I patients and 4 stage II patients. For stage I patients, EBUS-TBNA was significantly better (90.3%, p<0.001), but each modality showed 100% accuracy for stage II patients. CONCLUSION: It is recommended that EBUS-TBNA is added to the conventional diagnostic modalities for patients with suspicious stage I sarcoidosis on chest roentgenogram. PMID- 19674880 TI - Role of cyclin inhibitor protein p21 in the inhibition of HCT116 human colon cancer cell proliferation by American ginseng (Panax quinquefolius) and its constituents. AB - American ginseng and its ginsenoside constituents have been shown to exert anti cancer effects although the mechanism of action remains unclear. The present study determined the effects of water-extracted ginseng (GE) or its ginsenoside (GF) and polysaccharide (PS) fractions on the proliferation of human colon cancer cells and examined the role of p21 in mediating these effects using wild-type and p21-/- HCT116 human colon carcinoma cells. Proliferation was inhibited by GE, GF, and PS in wild-type and p21-/- cells, and the p21-/- cells were more sensitive to these treatments. Wild type cells treated with GE were arrested in the G0/G1 phase of the cell cycle and the expression of p53 and p21 proteins was increased while phospho-MEK levels decreased. In contrast, cells deficient in p21 displayed reduced cell viability, elevated number of dead cells, and increased expression of Bax and cleaved caspase-3 proteins. Both polysaccharides and ginsenosides appear to be responsible for the anti-proliferative and proapoptotic effects of GE. This study suggests that p21 functions to arrest HCT116 wild-type cells treated with GE, while p21-deficient cells undergo cell death in a ginseng constituent-dependent manner. PMID- 19674884 TI - Pyrolysis of the tetra pak. AB - This study deals with pyrolysis of tetra pak which is widely used as an aseptic beverage packaging material. Pyrolysis experiments were carried out under inert atmosphere in a batch reactor at different temperatures and by different pyrolysis modes (one- and two-step). The yields of char, liquid and gas were quantified. Pyrolysis liquids produced were collected as three separate phases; aqueous phase, tar and polyethylene wax. Characterization of wax and the determination of the total amount of phenols in aqueous phase were performed. Chemical compositions of gas and char products relevant to fuel applications were determined. Pure aluminum can be also recovered by pyrolysis. PMID- 19674886 TI - Highly sensitive capacitive immunosensor based on porous silicon-polyaniline structure: Bias dependence on specificity. AB - Porous silicon/polyaniline (PS/PANI) heterostructure has been prepared electrochemically which provides label free, real-time electrical detection with high sensitivity for the specific model biomolecules (mouse IgG/goat antimouse IgG). The sensor structures based on PS/PANI were prepared easily by directly immobilizing the biomolecule through gluteraldehyde coupler. These structures were used to detect specific biomolecule down to at least a picomolar concentration range and were found to be reusable. High sensitivity and easy processability of PS/PANI structures could be understood by the change in surface charge of the silicon channels in the PS structure that occurs by PANI deposition whilst providing covalent binding with biomolecules with its naturally occurring NH groups. The space charge layer in the PS/PANI channels get further modified by the surface charge change created by the specific analyte-antibody binding giving rise to high sensitivity. It was found that a substrate bias in the accumulation region of porous silicon gives the optimum sensitivity and specificity. PMID- 19674883 TI - Plakophilins: multifunctional scaffolds for adhesion and signaling. AB - Armadillo family proteins known as plakophilins have been characterized as structural components of desmosomes that stabilize and strengthen adhesion by enhancing attachments with the intermediate filament cytoskeleton. However, plakophilins and their close relatives are emerging as versatile scaffolds for multiple signaling and metabolic processes that not only facilitate junction dynamics but also more globally regulate diverse cellular activities. While perturbation of plakophilin functions contribute to inherited diseases and cancer pathogenesis, the functional significance of the multiple PKP isoforms and the mechanisms by which their behaviors are regulated remain to be elucidated. PMID- 19674885 TI - Spectral-domain optical coherence phase microscopy for label-free multiplexed protein microarray assay. AB - Quantitative measurement of affinities and kinetics of various biomolecular interactions such as protein-protein, protein-DNA and receptor-ligand is central to our understanding of basic molecular and cellular functions and is useful for therapeutic evaluation. Here, we describe a laser-scanning quantitative imaging method, referred to as spectral-domain optical coherence phase microscopy, as an optical platform for label-free detection of biomolecular interactions. The instrument is based on a confocal interferometric microscope that enables depth resolved quantitative phase measurements on sensor surface with high spatial resolution and phase stability. We demonstrate picogram per square millimeter surface mass sensitivity, and show its sensing capability by presenting static and dynamic detection of multiplexed protein microarray as immobilized antigens capture their corresponding antibodies. PMID- 19674887 TI - A one-compartment fructose/air biological fuel cell based on direct electron transfer. AB - The construction and characterization of a one-compartment fructose/air biological fuel cell (BFC) based on direct electron transfer is reported. The BFC employs bilirubin oxidase and d-fructose dehydrogenase adsorbed on a cellulose multiwall carbon nanotube (MWCNT) matrix, reconstituted with an ionic liquid, as the biocathode and the bioanode for oxygen reduction and fructose oxidation reactions, respectively. The performance of the bioelectrode was investigated by chronoamperometric and cyclic voltammetric techniques in a standard three electrode cell, and the polarization and long-term stability of the BFC was tested by potentiostatic discharge. An open circuit voltage of 663 mV and a maximum power density of 126 microWcm(-2) were obtained in buffer at pH 5.0. Using this regenerated cellulose-MWCNT matrix as the immobilization platform, this BFC has shown a relatively high performance and long-term stability compared with previous studies. PMID- 19674888 TI - Optical biosensors for environmental monitoring based on computational and biotechnological tools for engineering the photosynthetic D1 protein of Chlamydomonas reinhardtii. AB - Homology-based protein modelling and computational screening followed by virtual mutagenesis analyses were used to identify functional amino acids in the D1 protein of the photosynthetic electron transfer chain interacting with herbicides. A library of functional mutations in the unicellular green alga Chlamydomonas reinhardtii for preparing biomediators was built and their interactions with herbicides were calculated. D1 proteins giving the lowest and highest binding energy with herbicides were considered as suitable for preparing the environmental biosensors for detecting specific herbicide classes. Arising from the results of theoretical calculations, three mutants were prepared by site directed mutagenesis and characterized by fluorescence analysis. Their adsorption and selective recognition ability were studied by an equilibrium-adsorption method. The S268C and S264K biomediators showed high sensitivity and resistance, respectively, to both triazine and urea classes of herbicides. When immobilized on a silicon septum, the biomediators were found to be highly stable, remaining so for at least 1-month at room temperature. The fluorescence properties were exploited and a reusable and portable multiarray optical biosensor for environmental monitoring was developed with limits of detection between 0.8 x 10( 11) and 3.0 x 10(-9), depending on the target analyte. In addition, biomediator regeneration without obvious deterioration in performance was demonstrated. PMID- 19674889 TI - Sequence-specific DNA detection by using biocatalyzed electrochemiluminescence and non-fouling surfaces. AB - We herein report a DNA sensor for the sequence-specific DNA detection by using glucose oxidase-based biocatalyzed electrochemiluminescence and non-fouling surfaces. In this design, a glucose oxidase-labeled sandwich-type DNA sensor was built on a non-fouling surfaces made of a mixed self-assembled monolayers (SAMs) incorporating thiolated oligonucleotides and oligo(ethylene glycol) (OEG) thiols (SH-DNA/OEG). The sequence-specific DNA sensing was accomplished by the electrochemiluminesce (ECL) signal of luminol with the in-situ generated H(2)O(2). The protein-resistant non-fouling surfaces significantly suppressed the non-specific adsorption of enzyme label on electrode and reduced the background noise of this sensor. This sensor was able to detect as little as 1 pM of target DNA in pure buffer matrix. In complicated biological fluids such as human serum, this non-fouling platform-based sensor also revealed superior performance over conventional sandwich-type DNA sensors with mercaptohexanol (MCH)-coated surfaces. This strategy combines the high sensitivity of enzymatic amplified ECL and protein-resistant non-fouling platform, which is expected to be extended to a range of biological detection. PMID- 19674890 TI - Representations in auditory cortex. AB - How does auditory cortex represent auditory stimuli, and how do these representations contribute to behavior? Recent experimental evidence suggests that activity in auditory cortex consists of sparse and highly synchronized volleys of activity, observed both in anesthetized and awake animals. Many neurons are capable of remarkably precise activity with very low jitter or spike count variability. Most importantly, animals are capable of exploiting such precise neuronal activity in making sensory decisions. Whether the ability of auditory cortex to exploit fine temporal differences in cortical activity is unique to auditory modality, or represents a general strategy used by cortical circuits remains an open question. PMID- 19674893 TI - Quantitative analysis of sugars in wood hydrolyzates with 1H NMR during the autohydrolysis of hardwoods. AB - The focus of this work was to determine the utility of (1)H NMR spectroscopy in the quantification of sugars resulting from the solubilization of hemicelluloses during the autohydrolysis of hardwoods and the use of this technique to evaluate the kinetics of this process over a range of temperatures and times. Yields of residual xylan, xylooligomers, xylose, glucose, and the degraded products of sugars, i.e., furfural and HMF (5-hydroxymethyl furfural), were determined. The monosaccharide and oligomer contents were quantified with a recently developed high resolution (1)H NMR spectroscopic analysis. This method provided precise measurement of the residual xylan and cellulose remaining in the extracted wood samples and xylose and glucose in the hydrolyzates. NMR was found to exhibit good repeatability and provided carbohydrate compositional results comparable to published methods for sugar maple and aspen woods. PMID- 19674891 TI - Cell-type identity: a key to unlocking the function of neocortical circuits. AB - A central tenet of neuroscience is that the precise patterns of connectivity among neurons in a given brain area underlie its function. However, assigning any aspect of perception or behavior to the wiring of local circuits has been challenging. Here, we review recent work in sensory neocortex that demonstrates the power of identifying specific cell types when investigating the functional organization of brain circuits. These studies indicate that knowing the identity of both the presynaptic and postsynaptic cell type is key when analyzing neocortical circuits. Furthermore, identifying the circuit organization of particular cell types in the neocortex allows the recording and manipulation of each cell type's activity and the direct testing of its functional role in perception and behavior. PMID- 19674892 TI - Linking peripheral taste processes to behavior. AB - The act of eating and drinking brings food-related chemicals into contact with taste cells. Activation of these taste cells, in turn, engages neural circuits in the central nervous system that help animals identify foods and fluids, determine what and how much to eat, and prepare the body for digestion and assimilation. Analytically speaking, these neural processes can be divided into at least three categories: stimulus identification, ingestive motivation, and digestive preparation. This review will discuss recent advances in peripheral gustatory mechanisms, primarily from rodent models, in the context of these three major categories of taste function. PMID- 19674894 TI - Discovery and in vitro and in vivo profiles of 4-fluoro-N-[4-[6 (isopropylamino)pyrimidin-4-yl]-1,3-thiazol-2-yl]-N-methylbenzamide as novel class of an orally active metabotropic glutamate receptor 1 (mGluR1) antagonist. AB - We identified 4-fluoro-N-[4-[6-(isopropylamino)pyrimidin-4-yl]-1,3-thiazol-2-yl] N-methylbenzamide 27 as a potent mGluR1 antagonist. The compound possessed excellent subtype selectivity and good PK profile in rats. It also demonstrated relatively potent antipsychotic-like effects in several animal models. Suitable for development as a PET tracer, compound 27 would have great potential for elucidation of mGluR1 functions in human. PMID- 19674895 TI - Synthesis and biological evaluation of quinic acid derivatives as anti inflammatory agents. AB - Quinic acid (QA) esters found in hot water extracts of Uncaria tomentosa (a.k.a. cat's claw) exert anti-inflammatory activity through mechanisms involving inhibition of the pro-inflammatory transcription factor nuclear factor kappa B (NF-kappaB). Herein, we describe the synthesis and biological testing of novel QA derivatives. Inhibition of NF-kappaB was assessed using A549 (Type II alveolar epithelial-like) cells that stably express a secreted alkaline phosphatase (SEAP) reporter driven by an NF-kappaB response element. A549-NF-kappaB cells were stimulated with TNF-alpha (10 ng/mL) in the presence or absence of QA derivative for 18 hours followed by measurement of SEAP activity. Amide substitution at the carboxylic acid position yielded potent inhibitors of NF-kappaB. A variety of modifications to the amide substitution were tolerated with the N-propyl amide derivative being the most potent. Further examination of the SAR demonstrated that acetylation of the hydroxyl groups reduced NF-kappaB inhibitory activity. QA amide derivatives lacked anti-oxidant activity and were found to be neither anti proliferative nor cytotoxic at concentrations up to 100 microM. In conclusion, we have discovered a novel series of non-toxic QA amides that potently inhibit NF kappaB, despite their lack of anti-oxidant activity. Mechanistic studies and pre clinical efficacy studies in various inflammatory animal models are on-going. PMID- 19674896 TI - Discovery of a novel class of isoxazoline voltage gated sodium channel blockers. AB - Analogs of the previously reported voltage gated sodium channel blocker CDA54 were prepared in which one of the amide functions was replaced with aromatic and non-aromatic heterocycles. Replacement of the amide with an aromatic heterocycle resulted in significant loss of sodium channel blocking activity, while non aromatic heterocycle replacements were well tolerated. PMID- 19674897 TI - From natural products to achiral drug prototypes: potent thrombin inhibitors based on P2/P3 dihydropyrid-2-one core motifs. AB - A series of dihydropyrid-2-ones was synthesized and tested for inhibitory activity against serine protease enzymes. Moderate to low nanomolar inhibitory activities were obtained against thrombin and excellent selectivity against trypsin was observed. PMID- 19674898 TI - Spiropiperidine CCR5 antagonists. AB - A novel series of CCR5 antagonists has been identified, utilizing leads from high throughput screening which were further modified based on insights from competitor molecules. Lead optimization was pursued by balancing opposing trends of metabolic stability and potency. Selective and potent analogs with good pharmacokinetic properties were successfully developed. PMID- 19674899 TI - Disubstituted pyrimidines as Lck inhibitors. AB - We have developed a family of 4-benzimidazolyl-N-piperazinethyl-pyrimidin-2 amines that are subnanomolar inhibitors of Lck. A subset of these Lck inhibitors, with heterocyclic substituents at the benzimidazole C5, are also low-nanomolar inhibitors of cellular IL2 release. PMID- 19674901 TI - Comparing two devices of suspended treadmill walking by varying body unloading and Froude number. AB - In rehabilitation, treadmill walking with body weight support is commonly used to reduce loads on lower extremities. (1) However, gait pattern alterations during unloading at constant Froude number are infrequently reported. (2) Furthermore, differences between two common devices for unloading are not well known. Therefore, we investigated two devices; a waist-high chamber with increased pressure called Lower Body Positive Pressure (LBPP), and a harness system (LiteGait), considered a standard system for unloading the lower body. Four gait parameters (cadence, normalized stride length, duty factor, and leg angle at touch down), heart rate, and comfort level were monitored in 12 healthy volunteers. Subjects walked at three body weight (BW) conditions (100%, 66%, and 33% BW) and three Froude numbers (Fr), which refer to a dimensionless speed reflecting slow walking (Fr=0.09), comfortable walking (Fr=0.25), and walk-run transition (Fr=0.5). Absolute treadmill speed was determined using Froude numbers reflecting dynamically similar motions during unloading. We found that (1) the normal gait pattern is altered during unloading at a constant Froude number. In rehabilitation, physical therapists should be aware that normal gait pattern may not need to be maintained during unloaded treadmill walking. (2) Gait parameters were not different when comparing LBPP to harness supported walking. However, heart rate was lower and comfort higher during unloaded LBPP ambulation compared to suspended harness walking. Therefore, suspended LBPP walking may be more appropriate for patients with cardiovascular disease and for conditions at high unloading. PMID- 19674900 TI - Effects of physical guidance on short-term learning of walking on a narrow beam. AB - Physical guidance is often used in rehabilitation when teaching patients to re learn movements. However, the effects of guidance on motor learning of complex skills, such as walking balance, are not clear. We tested four groups of healthy subjects that practiced walking on a narrow (1.27 cm) or wide (2.5 cm) treadmill mounted balance beam, with or without physical guidance. Assistance was given by springs attached to a hip belt that applied restoring forces towards beam center. Subjects were evaluated while walking unassisted before and after training by calculating the number of times subjects stepped off of the beam per minute of successful walking on the beam (Failures per Minute). Subjects in Unassisted groups had greater performance improvements in walking balance from pre to post compared to subjects in Assisted groups. During training, Unassisted groups had more Failures per Minute than Assisted groups. Performance improvements were smaller in Narrow Beam groups than in Wide Beam groups. The Unassisted-Wide and Assisted-Narrow groups had similar Failures per Minute during training, but the Unassisted-Wide group had much greater performance gains after training. These results suggest that physical assistance can hinder motor learning of walking balance, assistance appears less detrimental for more difficult tasks, and task specific dynamics are important to learning independent of error experience. PMID- 19674902 TI - Outer membrane translocation: numerical protein secretion nomenclature in question in mycobacteria. PMID- 19674903 TI - Special feature of mixed phosphotriester derivatives of cytarabine. AB - Despite the unquestionable therapeutic interest of bis(SATE) pronucleotides, a presystemic metabolism preventing the delivery of the prodrugs in target cancer cells or tumours may constitute a limitation to the in vivo development of such derivatives. In order to overcome these drawbacks several strategies have been envisaged and we report herein the application of the S-acyl-2-thioethyl (SATE) phenyl pronucleotide approach to the well-known cytotoxic nucleoside cytosine-1 beta-D-arabinofuranoside (cytarabine, araC). We describe modifications of the SATE moieties with the introduction of polar groups on the acyl residue, in order to study how these changes affect antitumoral activity and metabolic stability. Two different synthetic pathways were explored and lead to obtain the corresponding mixed derivatives in satisfactory yields. Cytotoxicity was studied in murine leukaemia cells L1210 as well as in cells derived from solid human tumours (Messa and MCF7). Biological evaluation of these compounds in cell culture experiments with nucleoside analogue-sensitive and resistant cell lines showed that the modified compounds were active at higher concentrations than unmodified cytarabine, yet were much able to partially reverse resistance due to deficient nucleoside transport or activation. These results can be correlated with an incomplete decomposition mechanism into the corresponding 5' mononucleotide. PMID- 19674904 TI - Synthesis and biological evaluation of asymmetric gramicidin S analogues containing modified D-phenylalanine residues. AB - The synthesis of new analogues of the cationic antimicrobial peptide gramicidin S, having a modified D-phenylalanine residue, their antibacterial properties against several gram positive and negative strains, as well as their hemolytic activity is reported. PMID- 19674905 TI - Synthesis and evaluation of bioactive naphthoquinones from the Brazilian medicinal plant, Tabebuia avellanedae. AB - A series of naphthoquinones based on the naphtho[2,3-b]furan-4,9-dione skeleton such as (-)-5-hydroxy-2-(1'-hydoxyethyl)naphtho[2,3-b]furan-4,9-dione (1) and its positional isomer, (-)-8-hydroxy-2-(1'-hydoxyethyl)naphtho[2,3-b]furan-4,9-dione (2), which are secondary metabolites found in the inner bark of Tabebuia avellanedae, were stereoselectively synthesized and their biological activities were evaluated in conjunction with those of their corresponding enantiomers. Compound 1 exhibited potent antiproliferative effect against several human tumor cell lines, but its effect against some human normal cell lines was much lower than that of mitomycin. On the other hand, its enantiomer (R)-1 was less active toward the above tumor cell lines than 1. The antiproliferative effect of 2 against all tumor cell lines was significantly reduced. These results indicated the presence of the phenolic hydroxy group at C-5 is of great important for increasing antiproliferative effect. In addition, 1 also showed higher cancer chemopreventive activity than 2, while there were no significant differences between 1 and 2 in antimicrobial activity. Both compounds displayed modest antifungal and antibacterial activity (gram-positive bacteria), whereas they were inactive against gram-negative bacteria. PMID- 19674906 TI - Antioxidant efficacy of iridoid and phenylethanoid glycosides from the medicinal plant Teucrium chamaedris in cell-free systems. AB - Twelve glycosides, seven iridoids and five phenylethanoids, have been isolated from leaf and root methanolic extracts of Wall Germander (Teucrium chamaedrys), a Mediterranean species historically used as a medicinal plant. Among them, three iridoid and one phenylethanoid glycosides have been isolated and characterized for the first time. All of the structures have been elucidated on the basis of their spectral data, especially 1D and 2D NMR experiments. The antioxidative properties of pure metabolites, as well as of crude organic extracts of the plant, have been analyzed on the basis of their DPPH radical scavenging capability. The antioxidant capacity in cell-free systems of the isolated metabolites was carried out by measuring their capabilities to inhibit the synthesis of thiobarbituric acid reactive species in assay media using as oxidable substrates a vegetable fat and the pentose sugar 2-deoxyribose and to prevent oxidative damage of the hydrosoluble bovine serum albumin (BSA) protein. Phenylethanoid glycosides resulted efficacious DPPH radical, while iridoid glycosides prevent massively the 2-deoxyribose and BSA oxidations in assay media. PMID- 19674907 TI - Staurosporine tethered peptide ligands that target cAMP-dependent protein kinase (PKA): optimization and selectivity profiling. AB - We have recently developed a fragment based selection strategy for targeting kinases, where a small molecule warhead can be non-covalently tethered to a phage displayed library of peptides. This approach was applied to the conversion of the promiscuous kinase inhibitor, staurosporine, into a potent bivalent ligand for cAMP-dependent protein kinase (PKA). Herein we report a systematic evaluation of this new bivalent ligand (BL); (a) Lineweaver-Burke analysis revealed that the BL, unlike substrate-based bivalent kinase inhibitors, displayed non-competitive inhibition with respect to the peptide substrate, suggesting an allosteric mechanism of action; (b) linker optimization of the BL, afforded one of the most potent, sub-nanomolar, inhibitors of PKA reported to date; (c) the BL was found to be modular, where attachment of active site targeted small molecule warheads in lieu of staurosporine could achieve similar gains in affinity; and (d) profiling studies of both the staurosporine derivative and the BL (amide isostere) against a panel of 90 kinases revealed almost unique enhancement in selectivity against PKA (>5-fold) compared to the starting staurosporine derivative. These combined results provide new insights for BL discovery, which has the potential to provide guidance toward the development of kinase selective reagents while uncovering new allosteric sites on kinases for therapeutic targeting. PMID- 19674908 TI - Synthesis of pentopyranosyl-containing thiodisaccharides. Inhibitory activity against beta-glycosidases. AB - Beta-(1-->4)-thiodisaccharides formed by a pentopyranose unit as reducing or non reducing end have been synthesized using a sugar enone derived from a hexose or pentose as Michael acceptor of a 1-thiopentopyranose or 1-thiohexopyranose derivatives. Thus, 2-propyl per-O-acetyl-3-deoxy-4-S-(beta-D-Xylp)-4 thiohexopyranosid-2-ulose (3) and benzyl per-O-acetyl-3-deoxy-4-S-(beta-D-Galp)-4 thiopentopyranosid-2-ulose (11) were obtained in almost quantitative yields. The carbonyl function of these uloses was reduced with NaBH(4) or K-Selectride, and the stereochemical course of the reduction was highly dependent on the reaction temperature, reducing agent and solvent. Unexpectedly, reduction of 3 with NaBH(4)-THF at 0 degrees C gave a 3-deoxy-4-S-(beta-D-Xylp)-4-thio-alpha-D-ribo hexopyranoside derivative (6) as major product (74% yield), with isomerization of the sulfur-substituted C-4 stereocenter of the pyranone. Reduction of 11 gave always as major product the benzyl 3-deoxy-4-S-(Galp)-4-thio-beta-D-threo pentopyranoside derivative 14, which was the only product isolated (80% yield) in the reduction with K-Selectride in THF at -78 degrees C. Deprotection of 14 and its epimer at C-2 (13) afforded, respectively the free thiodisaccharides 19 and 18. They displayed strong inhibitory activity against the beta-galactosidase from Escherichia coli. Thus, compound 18 proved to be a non-competitive inhibitor of the enzyme (K(i)=0.80 mM), whereas 19 was a mixed-type inhibitor (K(i)=32 microM). PMID- 19674909 TI - Novel semisynthetic derivatives of betulin and betulinic acid with cytotoxic activity. AB - A series of new imidazole carboxylic esters (carbamates) and N-acylimidazole derivatives of betulin and betulinic acid (14-29) have been synthesized. The new compounds were screened for in vitro cytotoxicity activity against human cancer cell lines HepG2, Jurkat and HeLa. A number of compounds have shown IC(50) values lower than 2 microM against the cancer cell lines tested and the vast majority has shown a better cytotoxicity profile than betulinic acid, including the betulin derivatives. N-Acylimidazole derivatives 26 and 27 (IC(50) 0.8 and 1.7 microM in HepG2 cells) and the C-3 carbamate derivative 16 (IC(50) 2.0 microM in HepG2 cells) were the most promising compounds. Based on the observed cytotoxicity, structure-activity relationships have been established. PMID- 19674910 TI - Inhibitory effect of beta-diketones and their metal complexes on TNF-alpha induced expression of ICAM-1 on human endothelial cells. AB - Recent reports show that the natural beta-diketone curcumin displays important biological properties regarding the intercellular adhesion molecule-1 (ICAM-1), which plays a critical role in the immune responses and inflammation. In this study the ICAM-1 inhibitory activity of beta-diketone compounds, which are curcumin models lacking aromatic peripheral hydroxyl and methoxy groups, along with some metal derivatives is investigated. Beta-diketones are systematically more active than metal complexes and the best obtained inhibition is 75% for both groups. The best inhibitors are 4-benzoyl-3-methyl-1-phenyl-pyrazol-5-one (HQ(Ph)) among the ligands, and sodium benzoylacetonato among metal derivatives. These results appear in line with the reported antitumor activity of related species. Since 4-acyl-5-pyrazolones posses four tautomeric forms, those corresponding to HQ(Ph) were investigated using density functional theory. Docking of all HQ(Ph) tautomers on ICAM-1 protein was performed suggesting one keto-enol form favored to act in biological environment. PMID- 19674911 TI - Histochemical technique for the detection of chloride cells in fish. AB - Chloride cells are responsible for ionic exchanges in the fish gill. These cells have been widely studied, considering its importance in vital functions of the gill, and because they proliferate when exposed to unfavorable environments. One of the main characteristic of these cells is an acidic cytoplasm, which has been used for identification through histochemical techniques with dyes such as Toluidine Blue and Hematoxylin and Eosin. However, these techniques can be problematic, since epithelial cells can, in certain situations, acquire acidic characteristics similar to those of chloride cells, thus staining in a similar way. Among other functions, chloride cells play a role in calcium uptake from the environment, and therefore have a high concentration of this element. Based on this information, this study aims at developing a specific protocol for the identification of chloride cells. With this purpose, the Von Kossa method specific for calcium was used combined with Hematoxylin counterstaining. Chloride cells had cytoplasm slightly stained with Hematoxylin and the presence of dark stained granules dispersed in the cytoplasm resulted from the Von Kossa reaction due to the calcium present in these cells. This was not found in any other gill cell. Thus, the technique used in this study was specific and efficient to identify chloride cells in fish gills. PMID- 19674912 TI - In situ study of the metal-insulator transition in VO2 by EELS and ab initio calculations. AB - Changes in the dielectric properties during the thermochromic transition of commercial VO(2) powders were determined in situ, by analyzing the low-loss region of the electron energy-loss spectroscopy (EELS) spectra in a transmission electron microscope at room temperature (insulator phase) and 100 degrees C (metallic phase). A comparison of experimental EELS spectra and ab initio density functional theory calculations (WIEN2k code) within the generalized gradient approximation (GGA) is presented. A characteristic peak around 5.6 eV appears in the energy-loss function in metallic phase, which is absent in insulator phase. The origin of the characteristic peak is analyzed by means of energy-band structure calculations. PMID- 19674913 TI - Daunorubicin and doxorubicin inhibit the [(11)C]choline accumulation in cancer cells. AB - We studied how very short (10-40min) incubation with anthracycline derivatives modifies the accumulation of PET tumor-diagnostic radiotracers in cancer cells. The human ovarian A2780 and A2780AD, human B lymphoid JY, human epidermoid KB-3-1 and KB-V-1, and smooth muscle DDT1 MF-2 cells were pre-incubated with daunorubicin and doxorubicin, and the uptake of [(18)F]FDG and [(11)C]choline was measured. Anthracycline treatment decreased remarkably the [(11)C]choline accumulation in a concentration dependent manner, while it did not modify significantly the [(18)F]FDG uptake of the cells. PMID- 19674914 TI - The role of Cone beam CT in the evaluation and management of a family with Gardner's syndrome. AB - Gardner's syndrome (GS) is a hereditary autosomal dominant disease of the colon that presents with extra-colonic manifestations such as osteomas, skin lesions and dental abnormalities. Osteomas are commonly found in the skull, jaws and the paranasal sinuses. We present a family of four sisters affected with GS with a wide range of anomalies. The role of Cone beam computed tomography (CBCT) in the early detection and evaluation of osteomas and dental anomalies with precise assessment of their relationship to adjacent anatomic structures is described here in detail. The careful interpretation of CBCT may be of a great value in surgical and orthodontic treatment planning in the presence of jaw lesions. Management of dental problems in GS may be challenging due to the presence of odontomas and increased bone density. A multidisciplinary approach in the management of GS can achieve the best treatment results. PMID- 19674915 TI - Fast-track video-assisted bullectomy and pleurectomy for pneumothorax: initial experience and description of technique. AB - OBJECTIVE: Pleurectomy+/-bullectomy by video-assisted thoracoscopic surgery (VATS) is an established surgical procedure for pneumothorax. Early ambulation and discharge should be a reasonable goal. This study explores the feasibility of day-case surgery and identifies the obstacles requiring further work to facilitate day-case pneumothorax surgery. METHODS: Between June 2007 and May 2008, 16 consecutive patients underwent video-assisted thoracoscopic surgery bullectomy+/-pleurectomy (under the care of a single surgeon) with immediate connection to an ambulatory drainage system in the theatre following surgery. Analgesia comprised temporary paravertebral with early conversion to oral opiate+/-paracetamol. There were 13 males (81%), average age 23 (range: 17-29) years, and three females (19%), average age 35 (range: 22-46) years. Twelve patients (75%) had left-sided disease, of which nine (56%) underwent elective surgery. All patients had previously suffered at least one primary spontaneous pneumothorax. Patients with probable secondary pneumothorax were excluded from the study. Length of stay (LOS) was compared with a control group of patients conventionally treated prior to the study. RESULTS: In 13 patients (81%), early discharge was achieved 1 (range: 1-2 days) day post-op, whilst connected to an ambulatory drainage system. In three patients, early discharge was not achieved. One of these patients had the chest drain removed prematurely and remained an inpatient for 3 days with aspiration and observation for a small pneumothorax. The two remaining patients required extended inpatient admissions due to postoperative non-surgical complications. In the 13 patients discharged immediately, the time to drain removal (in clinic) was electively 7 days (range: 2-11 days). Two patients required re-admission: one for contralateral spontaneous pneumothorax and another for an ipsilateral basal pneumothorax treated with a drain. CONCLUSION: We have shown early discharge with ongoing ambulatory drainage following VATS pleurectomy+/-bullectomy in patients with primary pneumothorax to be feasible with paravertebral in the theatre and rapid conversion to oral analgesia. Patients managed intercostal drains at home. Limiting factors such as postoperative nausea and pain control usually can be sufficiently managed in the outpatient. Shorter stays may have a beneficial financial result. Long-term follow-up and a quantification of the patients experience is warranted. PMID- 19674916 TI - Postoperative lipid-lowering therapy and bioprosthesis structural valve deterioration: justification for a randomised trial? AB - OBJECTIVE: Bioprosthesis structural valve deterioration (SVD) is an incompletely understood process involving the accumulation of calcium and lipids. Whether this process could be delayed with lipid-lowering therapy (LLT) is currently unknown. The purpose of this observational study was to evaluate if an association exists between early LLT and a slowing of bioprosthesis SVD, with a view to designing a prospective trial. METHODS: We followed 1193 patients who underwent aortic valve replacement with contemporary bioprostheses between 1990 and 2006 (mean follow-up 4.5+/-3.1 years, maximum 17.3 years). Of these patients, 150 received LLT (including statins) early after surgery. Prosthetic valve haemodynamics on echocardiography and freedom from re-operation for SVD were compared between patients who did and did not receive postoperative LLT. RESULTS: After bioprosthetic implantation, the progression of peak and mean trans-prosthetic gradients during echocardiographic follow-up (mean 3.3 years) was equivalent between patients treated with and without LLT (peak increase: 0.9+/-7.7 vs 1.1+/ 10.9 mmHg, LLT vs no LLT, P=0.87; mean increase: 0.8+/-4.1 vs 0.2+/-5.9 mmHg, LLT vs no LLT, P=0.38). The annualised linear rate of gradient progression following valve replacement was also similar between groups (peak increase per year: 2.0+/ 12.1 vs 1.0+/-12.9 mmHg per year, LLT vs no LLT, P=0.52; mean increase per year: 0.5+/-2.2 vs 0.6+/-6.0 mmHg per year, LLT vs no LLT, P=0.94). The incidence of mild or greater aortic insufficiency on the most recent echocardiogram was comparable (16.3% vs 13.8%, LLT vs no LLT, P=0.44), and there was no difference in the 10-year freedom from re-operation for SVD between the two groups [98.9% (95% confidence interval (CI): 91.9%, 99.8%) vs 95.4% (95% CI 90.5%, 97.9%), LLT vs no LLT, P=0.72]. CONCLUSIONS: In this observational study, there was no association demonstrated between early postoperative LLT and a slowing of bioprosthesis SVD. With the excellent durability of bioprostheses in the current era, a prospective randomised trial of statin therapy to prevent bioprosthetic SVD does not appear to be justified, let alone feasible. PMID- 19674917 TI - Interaction of the cesium cation with mono-, di-, and tricarboxylic acids in the gas phase. A Cs+ affinity scale for cesium carboxylates ion pairs. AB - Humic substances (HS), including humic and fulvic acids, play a significant role in the fate of metals in soils. The interaction of metal cations with HS occurs predominantly through the ionized (anionic) acidic functions. In the context of the effect of HS on transport of radioactive cesium isotopes in soils, a study of the interaction between the cesium cation and model carboxylic acids was undertaken. Structure and energetics of the adducts formed between Cs+ and cesium carboxylate salts [Cs+RCOO-] were studied by the kinetic method and density functional theory (DFT). Clusters generated by electrospray ionization mass spectrometry from mixtures of a cesium salt (nitrate, iodide, trifluoroacetate) and carboxylic acids were quantitatively studied by CID. By combining the results of the kinetic method and the energetic data from DFT calculations, a scale of cesium cation affinity, CsCA, was built for 33 cesium carboxylates representing the first scale of cation affinity of molecular salts. The structural effects on the CsCA values are discussed. PMID- 19674918 TI - Three-dimensional spine kinematics during multidirectional, target-directed trunk movement in sitting. AB - The current study provides a quantitative assessment of three-dimensional spine motion during target-directed trunk movements in sitting. Subjects sat on an elevated surface, without foot support, and targets were placed in five directions, at three subject-specific distances (based on trunk height). Subjects were asked to lean toward the target, touch it with their head, and return to upright sitting. A retro-reflective motion analysis system was used to measure spine motion, using three kinematic trunk models (1, 3 and 7 segments). Significant differences were noted in the total trunk motion measured between the models, as well as between target distances and directions. In the most segmented model, inter-segmental trunk motion was also found to differ between trunk levels, with complex interaction effects involving target distance and direction. These findings suggest that inter-segmental spine motion is complex, task dependent, and often unevenly distributed between spine levels, with motion patterns differing between subjects, even in the absence of pathology. Use of a multi-segmental model provides the most interpretable findings, allowing for differentiation of individual motion patterns of the spine. Such an approach may be beneficial to the understanding of movement-related spine pathologies. PMID- 19674919 TI - The Starling mechanism of the urinary bladder contractile function and the influence of hyperglycemia on diabetic rats. AB - Diabetic cystopathy is one of the common complications of diabetes. Bladder dysfunction in diabetes is attributable to diabetic neuropathy that induces sensory and autonomic nerve dysfunction. MATERIALS AND METHODS: In the present study, the contractile mechanism of the bladder was evaluated both with and without electrical stimulation in normal rats, streptozotocin (STZ)-induced diabetic rats, and diabetic rats with insulin treatment. RESULTS: Both the normal and diabetic rats had optimal capacity of bladder and optimal length of detrusor muscle strips. The peak values of the volume-pressure curves of the bladder and length-tension curves of detrusor muscle strips as well as the enhanced values after electrical stimulation in 6- and 10-week diabetic groups were lower than in the 6- and 10-week normal groups and insulin-treated groups. However, there was no significant difference in peak and enhanced values between normal rats and diabetic rats treated with insulin, indicating that voiding function was improved after insulin treatment. CONCLUSIONS: The contractile function of rat's bladder including normal rats, diabetic rats, and diabetic rats treated with insulin is similar to the 'Starling mechanism.' It can be impaired by hyperglycemia, and insulin treatment is helpful to restore this function. PMID- 19674920 TI - Retention rate of pregabalin in drug-resistant epilepsy: 1-year follow-up, single centre observation in 105 consecutive, adult patients. AB - OBJECTIVES: Pregabalin (PGB) is a newer antiepileptic drug (AED) licensed as add on treatment for partial epilepsy in adults. Efficacy and safety have been proven in several controlled clinical studies. These trials, however, only partially reflect clinical practice. Retention rate has been established as a marker for efficacy and safety of AEDs in long-term follow-up studies. METHODS: We evaluated the data of the first 105 patients treated with PGB at Bethel Epilepsy Centre, a tertiary referral centre for epilepsy. The patients were interviewed after 3, 6 and 12 months. RESULTS: 105 adult patients (aged 38+/-13 years) were treated with PGB, on average in combination with 2.1 AEDs (mean observation period 232 days). 76.2% had focal epilepsy, 19.0 multifocal epilepsy, and 3.8% epilepsy with both focal and generalised seizures. 40% continued PGB with the following outcome: 5.7% seizure-free for at least 1 month (4.8% for at least 3 months, 2.4% for at least 6 months; one of the seizure-free patients, however, had had epilepsy surgery during the observational period), 17.1% responders (> or =50% reduction of seizure frequency but not seizure-free), 13.3% with unchanged or increased seizure frequency. Reasons for withdrawal were lack of efficacy (47.6%) or side effects (12.7%). CONCLUSIONS: PGB is a new therapeutic option as add-on therapy for patients with highly refractory focal epilepsies although the therapeutic success that can be expected in this group of patients is limited. PMID- 19674922 TI - Hormonal influences of early postnatal indomethacin and ibuprofen in neonatal rats. AB - OBJECTIVE: Indomethacin and ibuprofen are administered to preterm neonates for symptomatic patent ductus arteriosus. The drugs suppress prostaglandins (PGs) which modulate growth and secretion of various hormones. We examined the hypothesis that early postnatal indomethacin and ibuprofen influence growth and GH-IGF-I-insulin and HPA axes in neonatal rats. DESIGN: Rat pups received IP injections of saline (Sal) on P1, P2, and P3; 10mg/kg ibuprofen on P1 followed by 5mg/kg on P2 and P3; or 0.2mg/kg indomethacin on P1 followed by 0.1mg/kg on P2 and P3. Serum and hepatic GH, GHBP and IGF-I; and serum corticosterone and insulin levels were determined. RESULTS: Ibuprofen suppressed somatic growth in the sucking rats, but the effect was transient, resolving by P14. Indomethacin had an opposite, latent effect on body weight and liver to body weight ratios in weanling rats. Both indomethacin and ibuprofen had profound hormonal effects that differed in magnitude and timing. Indomethacin resulted in a sustained elevation in corticosterone levels at P21, while ibuprofen increased serum and hepatic GH levels. Both drugs suppressed GHBP in serum at P7 and P14; and liver at P4 and P7, but a rebound increase in serum GHBP was noted at P21 with Ibuprofen only. Both drugs increased serum IGF-I at P7. The effect remained sustained with indomethacin. CONCLUSIONS: These results provide evidence for an involvement of PGs in the regulation of growth as well as the GH-IGF and HPA axes. Therefore, early postnatal exposure to PG inhibitors may further exacerbate postnatal growth restriction and ability to cope with stress. PMID- 19674921 TI - Physical and psychosocial functioning following motor vehicle trauma: relationships with chronic pain, posttraumatic stress, and medication use. AB - Chronic pain and PTSD are known to hold substantial comorbidity following traumatic injury. Although pharmacological agents have been examined in the treatment of pain and PTSD individually, little is known regarding the relationship of medication use with functioning in patients with comorbid conditions. This research examined the relationships of pain, PTSD, and medication use across physical and psychosocial functioning in patients with chronic pain following motor vehicle injury (N=234). Separate analyses were conducted for opioids, SSRIs, and sedative/anxiolytics, respectively. Several relevant effects were noted: (1) Pain evidenced strong associations with reduced functioning across both physical and psychosocial domains, (2) Opioid use held interactive relationships with PTSD across both functioning domains. Specifically, opioids were associated with greater physical impairment in patients without comorbid PTSD. Opioids also were related to greater psychosocial impairment in patients without PTSD while PTSD was associated with greater impairment in patients not using opioids, (3) Opioid use evidenced a marginal interaction with pain on psychosocial functioning. Opioids were associated with greater psychosocial impairment among patients with high-pain, and high-pain was associated with greater impairment among opioid users, (4) SSRIs held a marginal interaction with PTSD such that PTSD was related to poorer psychosocial functioning only among individuals not using an SSRI, and (5) Anxiolytic use evidenced a marginal interaction with PTSD on physical functioning although no between-group differences were noted. These data suggest that PTSD symptomology may be an important consideration in determining treatment modality for patients experiencing pain subsequent to traumatic injury. PMID- 19674923 TI - Rabies exposure in international travelers: do we miss the target? AB - BACKGROUND: Little data exist about the spatial distribution of the risk for travelers of being injured by a potentially rabid animal. METHODS: Over the last 14 years, animal-associated injuries in 424 international travelers presenting to a travel medicine clinic in Marseille, southern France, were investigated. RESULTS: The majority of cases were reported from North Africa (41.5%) and Asia (22.2%). Most countries where at-risk injuries occurred (Algeria, Morocco, Tunisia, Thailand, and Turkey) were those for which travelers do not usually seek advice at a specialized travel clinic, because these countries are not at risk for specific travel-associated diseases like malaria or yellow fever. The probability of travelers being attacked by each animal species varied significantly according to the destination country. Dogs were more frequently involved in Algeria, cats in Tunisia and the Middle East, and non-human primates in sub-Saharan Africa, Madagascar, and Asia. CONCLUSIONS: We suggest that rabies pre-exposure vaccination should be offered to individuals traveling regularly to North Africa to visit their relatives and who are at high risk of exposure to potentially rabid animal attacks. Pre-travel advice when addressing rabies prevention should consider the specific epidemiology of animal-related injuries in the traveled country, as well as the traveler's characteristics. Travelers should be advised about which species of animal are potentially aggressive in their destination country so that they can more easily avoid risk-contacts. PMID- 19674924 TI - Ultrasonic degradation of Rhodamine B in the presence of hydrogen peroxide and some metal oxide. AB - In this research, degradation of Rodamine B in the presence of (hydrogen peroxide), (hydrogen peroxide+ultrasound), (hydrogen peroxide+aluminum oxide), (hydrogen peroxide+aluminum oxide+ultrasound with different ultrasound power), (hydrogen peroxide+iron oxide) and (hydrogen peroxide+iron oxide+ultrasound with different ultrasound power) were investigated at 25 degrees C. The apparent rate constants for the examined systems were calculated by pseudo-first-order kinetics. The results indicate that the rate of degradation was accelerated by ultrasound. The rate of degradation was increased by increasing power ultrasound. The efficiency of the (hydrogen peroxide+iron oxide+ultrasound) system for degradation of Rodamine B was higher than the others examined. PMID- 19674925 TI - Investigation of spiral blood flow in a model of arterial stenosis. AB - The spiral component of blood flow has both beneficial and detrimental effects in human circulatory system [Stonebridge PA, Brophy CM. Spiral laminar flow in arteries? Lancet 1991; 338: 1360-1]. We investigate the effects of the spiral blood flow in a model of three-dimensional arterial stenosis with a 75% cross sectional area reduction at the centre by means of computational fluid dynamics (CFD) techniques. The standard k-omega model is employed for simulation of the blood flow for the Reynolds number of 500 and 1000. We find that for Re=500 the spiral component of the blood flow increases both the total pressure and velocity of the blood, and some significant differences are found between the wall shear stresses of the spiral and non-spiral induced flow downstream of the stenosis. The turbulent kinetic energy is reduced by the spiral flow as it induces the rotational stabilities in the forward flow. For Re=1000 the tangential component of the blood velocity is most influenced by the spiral speed, but the effect of the spiral flow on the centreline turbulent kinetic energy and shear stress is mild. The results of the effects of the spiral flow are discussed in the paper along with the relevant pathological issues. PMID- 19674926 TI - Modeling and simulation of flexible needles. AB - Needle insertion is performed in many clinical and therapeutic procedures. Tissue displacement and needle bending which result from needle-tissue interaction make accurate targeting difficult. For performing physicians to gain essential needle targeting skills, needle insertion simulators can be used for training. An accurate needle bending model is essential for such simulators. These bending models are also needed for needle path planning. In this paper, three different models are presented to simulate the deformations of a needle. The first two models use the finite element method and take the geometric nonlinearity into account. The third model is a series of rigid bars connected by angular springs. The models were compared to recorded deformations during experiments of applying lateral tip forces on a brachytherapy needle. The model parameters were identified and the simulation results were compared to the experimental data. The results show that the angular spring model, which is computationally the most efficient model, is also the most accurate in modeling the bending of the brachytherapy needle. PMID- 19674927 TI - The benefits of a standardized patient education program for patients with Parkinson's disease and their caregivers. AB - The Patient Education Program Parkinson (PEPP) is a standardized psychosocial intervention aiming at improving the health-related quality of life (Hr-Qol) of patients with Parkinson's disease (PD) and caregivers. A randomized controlled trial was performed to assess its effectiveness. Sixty-four PD patients and 46 caregivers were allocated to either the intervention group (PEPP) or the control group (usual care). The intervention consisted of eight weekly sessions of 90 minute duration. Assessments were performed on psychosocial problems (BELA-P/A k), Hr-Qol (PDQ-39/EQ-5D) and depression (SDS) at baseline and one week after the end of the PEPP. Participants rated their mood on a visual analogue scale before and after each session. A significant effect for the caregivers on psychosocial problems and need for help was found and a trend for significance for patients' quality of life. Patients' and caregivers' mood improved significantly after each session. This study provides indications that PD patients and caregivers benefit from the PEPP. PMID- 19674928 TI - A study of internal rotations and vibrational spectra of oxiranemethanol (glycidol). AB - The conformational stability and the C-O and O-H internal rotations in oxiranemethanol were investigated at the DFT-B3LYP/6-311 G**, MP2/6-311 G** and MP4(SDQ)/6-311 G** levels of theory. Three minima were predicted in the CCOH potential energy scans of the molecule to have relative energies of about 2 kcal/mol or less and all were calculated to have real frequencies upon full optimization of structural parameters at the DFT and the MP2 levels of calculations. The Cg1 (H bond inner) conformation was predicted to be the lowest energy conformation for oxiranemethanol in excellent agreement with an earlier microwave study. The equilibrium mixture was calculated from Gibb's free-energy changes to be about 79% Cg1, 17% G1g and 3% Gg1 at the B3LYP/6-311G** level and about 87% Cg1, 11% G1g and 2% Gg1 at the MP2/6-311 G** level for oxiranemethanol at 298.15 K. No conclusive evidence was obtained for the presence of high-energy form in the liquid phase of oxiranemethanol. The vibrational frequencies of oxiranemethanol in its three stable forms were computed at the B3LYP level and complete vibrational assignments were made for the lowest energy Cg1 form on basis of calculated and experimental data of the molecule. PMID- 19674929 TI - Biological potential study of metal complexes of sulphonylurea glibenclamide on the house fly, Musca domestica (Diptera-Muscidae): preparation, spectroscopic and thermal characterization. AB - The ligatation behaviour of sulphonylurea glibenclamide drug is studied in order to give an idea about its potentiality towards some transition metals in vitro systems. Metal complexes of glibenclamide (GCA; H(3)L) drug are prepared and characterized based on elemental analyses, IR, diffused reflectance, magnetic moment, molar conductance and thermal analysis (TG and DTG) techniques. From the elemental analyses data, the complexes are proposed to have the general formulae [M(H(3)L)Cl(n)(H(2)O)(m)].yH(2)O (where M=Cr(III) (n=3, m=1, y=3); Mn(II) (n=2, m=0, y=1); Fe(III) (n=3, m=1, y=0), Co(II) (n=2, m=2, y=0); Ni(II) (n=2, m=2, y=3); Cu(II) (n=2, m=2, y=2) and Zn(II) (n=2, m=0, y=0). The molar conductance data reveal that all the metal chelates are non-electrolytes. IR spectra show that GCA is coordinated to the metal ions in a neutral bidentate manner with OO donor sites of the amide-O and sulphone-O. From the magnetic and solid reflectance spectra, it is found that the geometrical structures of these complexes are octahedral except Mn(II) and Zn(II) complexes which have tetrahedral structure. The thermal behaviour of these chelates is studied using thermogravimetric analysis (TG and DTG) technique. The activation thermodynamic parameters are calculated using Coats-Redfern method. The GCA drug, in comparison to its metal complexes also is screened for its biological activity against house fly, Musca domestica (Diptera-Muscidae). Dose of 5 microg/insect of GCA is topically applied against 3 days old larval instar of M. domestica. Survival of pupal and adult stages has been affected by the complexes of GCA more than larval instars. Morphogenic abnormalities of larvae, pupae and adults are studied. On the other hand pupation and adult emergence program is deteriorated by the effect of different chemicals. PMID- 19674930 TI - The first detection of human bocavirus 2 infections in China. PMID- 19674931 TI - Barcoding of diatoms: nuclear encoded ITS revisited. AB - DNA-barcoding is based on the premise that the divergence of a small DNA fragment coincides with biological separation of species. If true, it offers an additional tool for worldwide consistent species recognition even in cases of semi-cryptic species. Our study includes 618 sequences representing 114 diatom species belonging to the two most species-rich classes of diatoms (Mediophyceae and Bacillariophyceae). A 99.5% success rate in separating biologically defined species and a 91% success rate in separating all species tested was obtained when using the proposed barcode starting at the 5' end of 5.8S and ending in the conserved motif of helix III of ITS2 (300 to 400 bp). Including the whole 5.8S+ITS2 region did not significantly improve species resolution. We tested our barcode on 17 unidentified, misidentified or contaminated strains derived mostly from a culture collection, and these were correctly flagged as erroneous by their ITS sequences. We conclude that the proposed barcode represents for the Mediophyceae and Bacillariophyceae a robust, economical, and rapid way to recognize and identify most species (when a reference sequence is available) that is as good as or better than other molecular markers thus far proposed. PMID- 19674932 TI - Structure of loricae and stalks of several bacterivorous chrysomonads (chrysophyceae): taxonomical importance and possible ecological significance. AB - Stalks and loricae of chrysomonads were studied by fluorescence microscopy employing Calcoflor White-Evans blue (CW-Eb) staining, and by uranyl acetate staining of dried, whole mount preparations for electron microscopy. These structures were composed of microfibrils approximately 4 nm in diameter embedded in a matrix. The organization of the loricae of Poterioochromonas malhamenesis, "Amimonas minuta", Poterioochromonas stipitata and Ochromonas gloeopara showed a similar structural plan, consisting of a foot, stalk and cup region that together resemble a wine glass. CW-Eb-stained, microfibrillar stalks were identified also in Paraphysomonas vestita, Anthophysa vegetans and "Felimonas flocculans". These results suggest that CW-Eb-stained structures composed of microfibrils approximately 4 nm in diameter may be more common in chrysomonads than previously recognized. In cultures, these structures participate in the formation of cell aggregates and attachment of cells to substrates, and thus may be of ecological importance. Additionally, non-siliceous, CW-Eb-stained cysts were identified for the first time in P. malhamensis. PMID- 19674933 TI - The environmental clade LKM11 and Rozella form the deepest branching clade of fungi. AB - Previous environmental surveys of eukaryotic diversity using the small subunit ribosomal RNA (SSU rRNA) gene have revealed many clone sequences that branch near the base of Fungi. In this work, we demonstrate that many of these sequences, including those of the environmental clade LKM11, form a monophyletic and strongly supported group that also includes two sequences derived from the parasitic genus Rozella. This novel clade, called here "Rozellida", is the deepest branch of true fungi so far identified, and appears to be extremely diverse in the environment. PMID- 19674934 TI - Functional characterization of the 5'-upstream region of MTT5 metallothionein gene from Tetrahymena thermophila. AB - Metallothioneins are ubiquitous small, cysteine-rich, metal-binding proteins that play important roles in intracellular metal homeostasis and detoxification. Very few data are available on the promoter region and the mechanism of metallothionein transcription in Protozoa. In this study, we focused on Tetrahymena thermophila MTT5 5'-flanking region. To define the sequence elements underlying the metal-responsiveness of this promoter, we constructed a series of deletions and mutations starting with a 1777 bp fragment immediately upstream of the start codon of MTT5. As a reporter gene we used the previously tested IAG52B surface antigen from the protozoan fish parasite Ichthyophthirius multifiliis. The results suggest that a region spanning between -300 bp and -274 bp, dubbed Tetrahymena thermophila Cadmium-Response-Element (TtCdRE), is necessary to elicit high-level expression of the transgene following induction with cadmium. This is the first demonstration by in vivo analyses of a regulatory element essential for Cd-mediated control of protozoan metallothionein gene expression, where the sequence GATA appears to be involved. PMID- 19674935 TI - Distress in patients undergoing allogeneic haematopoietic stem cell transplantation is correlated with distress in nurses. AB - PURPOSE: To date, no studies have reported on the relationship between the emotional distress of patients undergoing allogeneic haematopoietic stem cell transplantation (HSCT) and the distress of their nurses. METHODS: 113 patients rated their distress by means of a daily questionnaire during their inpatient hospitalisation for HSCT. At the same time, nurses were asked to assess their distress caused by the additional care needs and increased emotional demands placed on them by the patients. Surveys covered a treatment period from day -5 to day +29 post-HSCT. RESULTS: The correlation between the distress level of the patients and that of the nurses was r=0.40 (p<0.001). The partial coefficient of this correlation was r=0.43 (p<0.001) when an indicator of the physical state of the patients, as assessed by their treating physicians, was controlled. CONCLUSIONS: Distress, as experienced by patients and nurses, is positively correlated. We assume that such a relation holds beyond other individual, organisational, structural, and occupational influences that play decisive and limiting roles in the patient-nurse relationship. Reducing the distress of one of these parties could also positively affect the distress levels of the other party. Therefore, patient distress should be monitored since it may add to nurse distress. Types of patient interventions, as well as the people responsible for performing these interventions, must be discussed. On the other hand, interventions aimed at reducing nurse distress (regardless of the cause of the distress) could reduce patient distress and improve their satisfaction with the treatment, and their quality of life. PMID- 19674936 TI - Adjuvant therapy with oral sodium clodronate in locally advanced and metastatic prostate cancer: long-term overall survival results from the MRC PR04 and PR05 randomised controlled trials. AB - BACKGROUND: Bisphosphonates might modulate the development of symptomatic bone metastases in men with prostate cancer. The Medical Research Council (MRC) PR05 and PR04 randomised controlled trials assessed the use of sodium clodronate, an oral, first-generation bisphosphonate. We report the final analyses of long-term survival data with additional follow-up in both trials. METHODS: 311 men with metastatic disease were recruited to PR05 between 1994 and 1998, and 508 men with non-metastatic disease were recruited to PR04 from 1994 to 1997. All men were treated according to the recruiting site's standard practice at the time: for metastatic disease, all men were starting or responding to long-term hormone therapy; for non-metastatic disease, most men had radiotherapy, hormone therapy, or both. Men were randomly assigned to take four tablets per day of sodium clodronate (2080 mg) or matching placebo for up to 3 years (metastatic disease) or 5 years (non-metastatic). Long-term overall survival was assessed on an intention-to-treat basis in all men at sites in England and Wales using data from the National Health Service Information Centre, which held data for 278 of 311 men in the PR05 trial and 471 of 508 men in the PR04 trial. These studies are registered International Standardised Randomised Controlled Trials, numbers ISRCTN38477744 (PR05) and ISRCTN61384873 (PR04). FINDINGS: Of the 278 men with metastatic disease, 258 (93%) were reported to have died. Evidence of a benefit for those with metastatic disease from use of sodium clodronate compared with placebo was seen in overall survival (hazard ratio [HR] 0.77, 95% CI 0.60-0.98; p=0.032). Of the 471 men with non-metastatic disease, 281 (60%) were reported to have died, with no evidence of improvement in overall survival with clodronate compared with placebo (HR 1.12, 0.89-1.42; p=0.94). INTERPRETATION: Long-term data from these trials show that a first-generation bisphosphonate, sodium clodronate, improves overall survival in men with metastatic prostate cancer who are starting hormone therapy, but there is no evidence of an effect in men with non-metastatic prostate cancer. FUNDING: UK MRC; and an education grant and free drug from Roche Products Ltd. PMID- 19674937 TI - Novel cytokine-targeted therapies and intestinal inflammation. AB - Several cytokines have been identified as critical mediators of chronic inflammation in inflammatory bowel disease (IBD) and biological therapies that target these molecules have been developed during recent years. Thereby, anti-TNF agents have noticeably improved the treatment of patients with IBD in comparison to conventional therapy. Furthermore, initial clinical trials showed promising results with anti-IL-6 and anti-IL-12/IL-23 agents. In addition to these well known mediators of IBD, various novel cytokines have been described as critical during the pathogenesis of IBD in recent experimental studies and therapeutic targeting of these cytokines could provide new strategies for human disease. PMID- 19674938 TI - Commentary to "Controversies in the management of vesicoureteral reflux - the rationale for the RIVUR study": Urinary tract infections and vesicoureteral reflux in childen: What have we learned? PMID- 19674940 TI - Using interactional and linguistic analysis to distinguish between epileptic and psychogenic nonepileptic seizures: a prospective, blinded multirater study. AB - This study was carried out to test the suggestion that close interactional and linguistic examination of the communication between neurologists and patients during a first encounter can contribute to the differential diagnosis of epilepsy or psychogenic nonepileptic seizures. Twenty unselected patients admitted for video/EEG telemetry because of diagnostic uncertainty were included. Two linguists blinded to all medical data independently studied video recordings and transcripts of 25- to 35-minute interactions. They attempted to predict the medical diagnosis on the basis of qualitative assessments addressing 17 separate observations. They also used a diagnostic scoring aid (DSA) to convert their qualitative assessments into a simple numeric score. Using qualitative assessment, both linguists predicted 17 of 20 (85%) diagnoses (kappa=0.59). With the DSA, diagnoses were predicted with a sensitivity of 85.7% (71.4%) and a specificity of 84.6% (92.3%). This blinded, prospective multirater study confirms the diagnostic value of linguistic and interactional observations in the seizure clinic setting. PMID- 19674939 TI - Hippocampal volumetry and functional MRI of memory in temporal lobe epilepsy. AB - This study examined the utility of structural and functional MRI at 1.5 and 3T in the presurgical evaluation and prediction of postsurgical cognitive outcome in temporal lobe epilepsy (TLE). Forty-nine patients undergoing presurgical evaluation for temporal lobe (TL) resection and 25 control subjects were studied. Patients completed standard presurgical evaluations, including the intracarotid amobarbital test (IAT) and neuropsychological testing. During functional imaging, subjects performed a complex visual scene-encoding task. High-resolution structural MRI scans were used to quantify hippocampal volumes. Both structural and functional imaging successfully lateralized the seizure focus and correlated with IAT memory lateralization, with improvement for functional imaging at 3T as compared with 1.5 T. Ipsilateral structural and functional MRI data were related to cognitive outcome, and greater functional asymmetry was related to earlier age at onset. These findings support continued investigation of the utility of MRI and fMRI in the presurgical evaluation of TLE. PMID- 19674942 TI - Visceral afferents - determinants and modulation of excitability. AB - An essential property of visceral sensory afferents is to be able to alter their firing properties in response to changes in the microenvironment at the level of the sensory ending. Significant progress has been made in recent years in understanding the ionic mechanisms of the regulation of afferent neuronal excitability, and in identifying the mechanisms by which this can be altered. This article will review some of the recent developments in the state of knowledge regarding mechanisms of increased excitability after inflammation, and pharmacological modulation of excitability, concentrating on afferent nerves innervating the GI tract and urinary bladder. PMID- 19674941 TI - Gut chemosensing: interactions between gut endocrine cells and visceral afferents. AB - Chemosensing in the gastrointestinal tract is less well understood than many aspects of gut mechanosensitivity; however, it is important in the overall function of the GI tract and indeed the organism as a whole. Chemosensing in the gut represents a complex interplay between the function of enteroendocrine (EEC) cells and visceral (primarily vagal) afferent neurons. In this brief review, I will concentrate on a new data on endocrine cells in chemosensing in the GI tract, in particular on new findings on glucose-sensing by gut EEC cells and the importance of incretin peptides and vagal afferents in glucose homeostasis, on the role of G protein coupled receptors in gut chemosensing, and on the possibility that gut endocrine cells may be involved in the detection of a luminal constituent other than nutrients, the microbiota. The role of vagal afferent pathways as a downstream target of EEC cell products will be considered and, in particular, exciting new data on the plasticity of the vagal afferent pathway with respect to expression of receptors for GI hormones and how this may play a role in energy homeostasis will also be discussed. PMID- 19674943 TI - Highly improved electrooxidation of glucose at a nickel(II) oxide/multi-walled carbon nanotube modified glassy carbon electrode. AB - Electrochemical oxidation of glucose on a glassy carbon disc electrode modified with multi-walled carbon nanotubes and nickel(II) oxide (GC/MWCNT/NiO) was examined by cyclic voltammetry and chronoamperometry in alkaline aqueous solutions. The results were compared with those obtained on a nickel(II) oxide modified glassy carbon electrode (GC/NiO). Both electrodes conditioned by potential cycling in a limited potential range (0.1-0.6 V vs. Ag/AgCl) in 0.10 M NaOH solution. It was found that the multi-walled carbon nanotubes improve remarkably the reactivity of nickel(II) oxide for glucose oxidation. The GC/MWCNT/NiO electrode exhibited good linear behavior in the concentration range from 2.0x10(-4) mol/L to 1.2x10(-2) mol/L for the quantitative analysis of glucose with a limit of detection of 1.6x10(-4) mol/L (3sigma). The prepared electrode exhibits satisfactory stability and long life if stored at ambient conditions. Finally, it has been demonstrated that the proposed modified electrode can be successfully used for the assay of glucose in serum samples. PMID- 19674944 TI - Directly applicable microbial fuel cells in aeration tank for wastewater treatment. AB - The application of microbial fuel cell (MFC) for wastewater treatment is a promising strategy for the simultaneous treatment of pollutants and generation of electricity. However, for practical application, there are several limitations to the MFC that involve biological and engineering aspects. In this study, a single chambered MFC able to submerge into the aeration tank of the activated sludge process was developed to optimize the cell configuration and electrode materials. Among four MFCs with different electrode materials, the MFC with a graphite felt (GF) anode and a GF cathode showed the highest power density of 16.7 W m(-3) and the lowest internal resistance of 17 Omega. When the blower was stopped to evaluate the effect of mixing intensity, the concentration of dissolved oxygen nevertheless remained at 8 mg O2 L(-1), and the cell voltage of MFCs dropped rapidly and reached 30 mV. However, the cell voltage immediately returned to around 200 mV after the blowing of air. The MFCs with a GF cathode were sensitive to mixing intensity. At the very low concentration of 0.2 mg O2 L(-1), the cell voltage remained at a high level of 200 mV when the oxygen close to the cathode remained and mixing was sufficient. PMID- 19674945 TI - Assays for the quantification of melphalan and its hydrolysis products in human plasma by liquid chromatography-tandem mass spectrometry. AB - Two high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS) assays are described for the quantification of melphalan in human plasma. N phenyldiethanolamine was tested as internal standard. The first assay consisted of a protein precipitation by cold methanol and a reversed-phase HPLC whereas the second one was based on a solid phase extraction and a hydrophilic interaction chromatography. Both provided a very satisfactory mean extraction yield with a small volume of sample. The first method was simple, rapid and used as a routine assay. The second one was developed in order to determine melphalan hydrolysis products and to avoid scarce cases when interferences from biological matrix alter the quantification of melphalan using the first method. The two assays were linear and sensitive in the range of 1-500ng/mL for the first one and in a range of 25-2000ng/mL for the second one. Concentrations out of the range fixed with the first method were also validated. The procedure was reliable with precision and accuracy below 10%. All compounds were detected after positive mode electrospray ionization in selected reaction monitoring mode. These new analytical procedures were developed for melphalan pharmacokinetic studies or therapeutic drug monitoring. PMID- 19674946 TI - Social likeability, conformity, and body talk: Does fat talk have a normative rival in female body image conversations? AB - Fat talk, dialogues among women involving negative body-focused discussions, was studied as a function of conformity and social likeability through the use of four vignettes depicting young women in conversation. Using a 2 (body presentation style of the group: negative or positive)x2 (body presentation style of the target, Jenny: negative or positive) factorial design, 215 college women (92.1% non-Hispanic Caucasian) read one of four vignettes in a classroom setting and made ratings on a social likeability scale. Participants' personal ratings of Jenny's likeability were higher when she spoke positively about her body, whereas they expected the other group members in the vignette to like Jenny more when she conformed to the group's body presentation style. This study is the first to support two competing norms for women's body image-the existing norm to fat talk versus a newly documented norm that some women like others who express body acceptance. PMID- 19674947 TI - The Male Body Attitudes Scale: A confirmatory factor analysis with a sample of gay men. AB - The Male Body Attitudes Scale (MBAS; Tylka, Bergeron, & Schwartz, 2005) is a recently created instrument which assesses males' attitudes regarding their muscularity, body fat, and height. Although the MBAS was created via exploratory and confirmatory factor analyses, its factor structure has yet to be replicated with more diverse samples. The aim of the current study was to conduct a confirmatory factor analysis of the MBAS with a sample of gay men. Results from an online sample of 207 self-identified gay men revealed that the original three factor structure of the MBAS, as well as a two-factor structure, consisting of muscularity and body fat, displayed strong factorial validity. These findings suggest that either a two- or three-factor structure may be used when assessing body image concerns in gay men. PMID- 19674948 TI - New spatial measure for dispersed-dot halftoning assuring good point distribution in any density. AB - As a core component of dispersed-dot halftoning, this paper focuses on the definition of new measures for giving or measuring good point distributions in a plane. By defining good point distributions from a purely geometric viewpoint of circle packing, it is shown that the energy defined by a certain strong convex function satisfies the necessary conditions for obtaining good point distributions in any point density by minimizing the energy. The energy with such the conditions are mathematically plain and there are no obscure parameters. The theory is also significantly motivated by a requirement of the adjustability to discrete spaces, and it is shown that the conditions actually work well also in the spaces. As an application, by using technically simple methods, dispersed-dot halftone masks are designed and goodness of point distributions of masks are estimated. PMID- 19674950 TI - Granular neural networks and their development through context-based clustering and adjustable dimensionality of receptive fields. AB - In this study, we present a new architecture of a granular neural network and provide a comprehensive design methodology as well as elaborate on an algorithmic setup supporting its development. The proposed neural network relates to a broad category of radial basis function neural networks (RBFNNs) in the sense that its topology involves a collection of receptive fields. In contrast to the standard architectures encountered in RBFNNs, here we form individual receptive fields in subspaces of the original input space rather than in the entire input space. These subspaces could be different for different receptive fields. The architecture of the network is fully reflective of the structure encountered in the training data which are granulated with the aid of clustering techniques. More specifically, the output space is granulated with use of K-means clustering while the information granules in the multidimensional input space are formed by using the so-called context-based fuzzy C-means, which takes into account the structure being already formed in the output space. The innovative development facet of the network involves a dynamic reduction of dimensionality of the input space in which the information granules are formed in the subspace of the overall input space which is formed by selecting a suitable subset of input variables so that this subspace retains the structure of the entire space. As this search is of combinatorial character, we use the technique of genetic optimization [genetic algorithms (GAs), to be more specific] to determine the optimal input subspaces. A series of numeric studies exploiting synthetic data and data coming from the Machine Learning Repository, University of California at Irvine, provide a detailed insight into the nature of the algorithm and its parameters as well as offer some comparative analysis. PMID- 19674951 TI - Wavelet differential neural network observer. AB - State estimation for uncertain systems affected by external noises is an important problem in control theory. This paper deals with a state observation problem when the dynamic model of a plant contains uncertainties or it is completely unknown. Differential neural network (NN) approach is applied in this uninformative situation but with activation functions described by wavelets. A new learning law, containing an adaptive adjustment rate, is suggested to imply the stability condition for the free parameters of the observer. Nominal weights are adjusted during the preliminary training process using the least mean square (LMS) method. Lyapunov theory is used to obtain the upper bounds for the weights dynamics as well as for the mean squared estimation error. Two numeric examples illustrate this approach: first, a nonlinear electric system, governed by the Chua's equation and second the Lorentz oscillator. Both systems are assumed to be affected by external perturbations and their parameters are unknown. PMID- 19674952 TI - Computation of image spatial entropy using quadrilateral Markov random field. AB - Shannon entropy is a powerful tool in image analysis, but its reliable computation from image data faces an inherent dimensionality problem that calls for a low-dimensional and closed form model for the pixel value distributions. The most promising such models are Markovian, however, the conventional Markov random field is hampered by noncausality and its causal versions are also not free of difficulties. For example, the Markov mesh random field has its own limitations due to the strong diagonal dependency in its local neighboring system. A new model, named quadrilateral Markov random field (QMRF) is introduced in this paper in order to overcome these limitations. A property of QMRF with neighboring size of 2 is then used to decompose an image prior into a product of 2-D joint pdfs in which they are estimated using a joint histogram under the homogeneity assumption. In addition, the paper includes an extension of the introduced method to the computation of image spatial mutual information. Comparisons on synthesized images as well as two applications with real images are presented to motivate the developments in this paper and demonstrate the advantages in the performance of the introduced method over the existing ones. PMID- 19674953 TI - Robust color demosaicking with adaptation to varying spectral correlations. AB - Almost all existing color demosaicking algorithms for digital cameras are designed on the assumption of high correlation between red, green, blue (or some other primary color) bands. They exploit spectral correlations between the primary color bands to interpolate the missing color samples, but in areas of no or weak spectral correlations, these algorithms are prone to large interpolation errors. Such demosaicking errors are visually objectionable because they tend to correlate with object boundaries and edges. This paper proposes a remedy to the above problem that has long been overlooked in the literature. The main contribution of this work is a hybrid demosaicking approach that supplements an existing color demosaicking algorithm by combining its results with those of adaptive intraband interpolation. This is formulated as an optimal data fusion problem, and two solutions are proposed: one is based on linear minimum mean square estimation and the other based on support vector regression. Experimental results demonstrate that the new hybrid approach is more robust and eliminates the worst type of color artifacts of existing color demosaicking methods. PMID- 19674954 TI - Octahedral transforms for 3-D image processing. AB - The octahedral group is one of the finite subgroups of the rotation group in 3-D Euclidean space and a symmetry group of the cubic grid. Compression and filtering of 3-D volumes are given as application examples of its representation theory. We give an overview over the finite subgroups of the 3-D rotation group and their classification. We summarize properties of the octahedral group and basic results from its representation theory. Wide-sense stationary processes are processes with group theoretical symmetries whose principal components are closely related to the representation theory of their symmetry group. Linear filter systems are defined as projection operators and symmetry-based filter systems are generalizations of the Fourier transforms. The algorithms are implemented in Maple/Matlab functions and worksheets. In the experimental part, we use two publicly available MRI volumes. It is shown that the assumption of wide-sense stationarity is realistic and the true principal components of the correlation matrix are very well approximated by the group theoretically predicted structure. We illustrate the nature of the different types of filter systems, their invariance and transformation properties. Finally, we show how thresholding in the transform domain can be used in 3-D signal processing. PMID- 19674955 TI - A multiobjective optimization approach to obtain decision thresholds for distributed detection in wireless sensor networks. AB - For distributed detection in a wireless sensor network, sensors arrive at decisions about a specific event that are then sent to a central fusion center that makes global inference about the event. For such systems, the determination of the decision thresholds for local sensors is an essential task. In this paper, we study the distributed detection problem and evaluate the sensor thresholds by formulating and solving a multiobjective optimization problem, where the objectives are to minimize the probability of error and the total energy consumption of the network. The problem is investigated and solved for two types of fusion schemes: 1) parallel decision fusion and 2) serial decision fusion. The Pareto optimal solutions are obtained using two different multiobjective optimization techniques. The normal boundary intersection (NBI) method converts the multiobjective problem into a number of single objective-constrained subproblems, where each subproblem can be solved with appropriate optimization methods and nondominating sorting genetic algorithm-II (NSGA-II), which is a multiobjective evolutionary algorithm. In our simulations, NBI yielded better and evenly distributed Pareto optimal solutions in a shorter time as compared with NSGA-II. The simulation results show that, instead of only minimizing the probability of error, multiobjective optimization provides a number of design alternatives, which achieve significant energy savings at the cost of slightly increasing the best achievable decision error probability. The simulation results also show that the parallel fusion model achieves better error probability, but the serial fusion model is more efficient in terms of energy consumption. PMID- 19674956 TI - Feasibility and effectiveness of online physical activity advice based on a personal activity monitor: randomized controlled trial. AB - BACKGROUND: Inactive people are often not aware of the fact that they are insufficiently active. Providing insight into their actual physical activity (PA) levels may raise awareness and could, in combination with tailored PA advice, stimulate a physically active lifestyle. OBJECTIVE: This study evaluated the feasibility and effectiveness of a 3-month intervention in which Dutch office workers were provided with a personal activity monitor (PAM) coupled to simple and concise Web-based tailored PA advice (PAM COACH). METHOD: Participants were randomly assigned to the 3-month PAM intervention (n = 51) or received a single written information brochure with brief general PA recommendations (n = 51). Study outcome measures were changes in PA (recall of minutes per week spent on PA, as measured by the Activity Questionnaire for Adolescents and Adults), determinants of PA, aerobic fitness, and body composition. Follow-up measurements were performed immediately after the 3-month intervention and at 8-months, 5 months after the end of the 3-month intervention period. RESULTS: A total of 102 workers, 23 to 39 years old, completed the baseline measurement at the worksite. 48 completed the 3-month follow up and 38 the 8-month follow-up in the intervention group, 50 completed the 3-month follow up and 42 the 8-month follow up in the control group. 35 out of 48 (73%) participants in the PAM intervention group reported wearing the PAM regularly, and the PAM COACH was used almost once a week; 24 out of 46 (52%) PAM users set a personal goal, and 33 (72%) entered their favorite activities on the website. Main reasons for not using these items were lack of interest or not being able to find the item on the website. The majority of PAM users (34 out of 46, 74%) read the advice, of whom 14 (39%) found it unappealing. After the 3-month intervention, no significant intervention effect was observed (adjusted difference in min/week) for sedentary behavior (beta = 10, 95% CI = -435 to 455), light-intensity PA (beta = -129, 95% CI = -337 to 79), moderate-intensity PA (beta = -13, 95% CI = -89 to 63), vigorous intensity PA (beta= -6, 95% CI = -75 to 62), and moderate- to vigorous-intensity PA (beta = -23, 95% CI = -121 to 76). No significant intervention effect was observed in the PA outcomes at the 8-month follow-up. For the determinants of PA, aerobic fitness, and body composition, no statistically significant intervention effect was observed in the total study population immediately after the 3-month intervention or the 8-month follow-up. CONCLUSIONS: The intervention appeared to be easily applicable to real-life settings. The intervention was ineffective in improving PA behavior or its determinants in healthy office workers. More attention should have been given to the quality and appropriateness of the tailored advice. TRIAL REGISTRATION: International Standard Randomized Controlled Trial Number (ISRCTN): 93896459; http://www.controlled-trials.com/ISRCTN93896459/ (Archived by WebCite at http://www.webcitation.org/5iR3mf7ex). PMID- 19674957 TI - Nationwide implementation of Hello World: a Dutch email-based health promotion program for pregnant women. AB - BACKGROUND: In November 2006, an email-based health promotion program for pregnant women was implemented nationally in the Netherlands. The program consisted of emails containing quizzes with pregnancy-related questions tailored to the number of weeks of pregnancy. Emails were sent out once every 4 weeks, up to a maximum of nine emails. OBJECTIVES: The aims of the study were (1) to assess the recruitment of participants and their representativeness of the Dutch population and (2) to study differences in recruitment, program use, and program appreciation among women with different levels of education. METHODS: Data from 13,946 pregnant women who enrolled during the first year of the program were included. Upon registration, participants were asked how they found out about the program and subsequently received an email questionnaire to assess demographic, lifestyle, and Internet characteristics. Program use was tracked, and participants were classified into five user groups (inactive to very active). Program appreciation (low, intermediate, and high) was assessed twice with an email questionnaire that was sent after the woman had received her third and sixth quiz email. Information about pregnant women and their characteristics was obtained from Dutch registries to assess representativeness of the study population. RESULTS: About 8% of the pregnant women in the Netherlands enrolled in the program. Immigrants were underrepresented, and women with a low level of education seemed to be slightly underrepresented. Most women knew about the program from a promotional email sent by the organization (32%), followed by the Internet (22%) and midwives (16%). Women with little education were more often inactive users of the program than were highly educated women (15% vs 11%, P < .001), whereas highly educated women were more often very active users compared with women with little education (25% vs 20%, P< .001). However, women with less education were more likely than women with more education to have a high appreciation of the program after receiving three quiz emails (52% vs 44%, P = .001). CONCLUSIONS: In this real-life setting, pregnant women can be reached through an email-based health promotion program. Selective engagement by education level remains a challenge. PMID- 19674958 TI - What is my cancer risk? How internet-based cancer risk assessment tools communicate individualized risk estimates to the public: content analysis. AB - BACKGROUND: Internet-based cancer risk assessment tools have the potential to inform the public about cancer risk and promote risk-reducing behaviors. However, poorly communicated information on these websites may result in unintended adverse health outcomes. OBJECTIVE: This study examined whether: (1) Internet based cancer risk assessment tools use risk communication formats that facilitate comprehension and reduce bias (as identified by the empirical literature); (2) the use of these formats varies by website affiliation; and (3) the websites provided information necessary to evaluate the quality of the risk estimate. METHODS: A content analysis of Internet-based cancer risk assessment tools was conducted. The terms calculate cancer risk, cancer risk calculator, estimate cancer risk, assess cancer risk, and cancer risk assessment were searched using three search engines. We identified 47 risk assessment tools and coded each according to standardized criteria. We calculated simple frequencies on all coding categories and performed crosstabulations but did not conduct formal statistical analysis due to small cell sizes. RESULTS: Use of risk communication formats that facilitate comprehension and reduce bias varied widely (eg, 30% of websites [14/47] provided absolute and comparative risk information but 83% [39/47] provided safety messages). Use of formats that facilitate comprehension varied by website affiliation and communication strategy (eg, only 8.3% [1/12] websites affiliated with the health care industry provided absolute and comparative risk information, but 83% [5/6] of websites affiliated with a governmental organization did so). Only 53% (25/47) of websites provided information about the statistical model or the peer-reviewed literature that was used to calculate the risk estimate. CONCLUSION: Internet-based cancer risk assessment tools varied in their use of risk communication formats that facilitate comprehension and reduce bias. Formats that are difficult to understand may cause people to misperceive their cancer risk and consequently take inappropriate action. PMID- 19674959 TI - Mobile and fixed computer use by doctors and nurses on hospital wards: multi method study on the relationships between clinician role, clinical task, and device choice. AB - BACKGROUND: Selecting the right mix of stationary and mobile computing devices is a significant challenge for system planners and implementers. There is very limited research evidence upon which to base such decisions. OBJECTIVE: We aimed to investigate the relationships between clinician role, clinical task, and selection of a computer hardware device in hospital wards. METHODS: Twenty-seven nurses and eight doctors were observed for a total of 80 hours as they used a range of computing devices to access a computerized provider order entry system on two wards at a major Sydney teaching hospital. Observers used a checklist to record the clinical tasks completed, devices used, and location of the activities. Field notes were also documented during observations. Semi-structured interviews were conducted after observation sessions. Assessment of the physical attributes of three devices-stationary PCs, computers on wheels (COWs) and tablet PCs-was made. Two types of COWs were available on the wards: generic COWs (laptops mounted on trolleys) and ergonomic COWs (an integrated computer and cart device). Heuristic evaluation of the user interfaces was also carried out. RESULTS: The majority (93.1%) of observed nursing tasks were conducted using generic COWs. Most nursing tasks were performed in patients' rooms (57%) or in the corridors (36%), with a small percentage at a patient's bedside (5%). Most nursing tasks related to the preparation and administration of drugs. Doctors on ward rounds conducted 57.3% of observed clinical tasks on generic COWs and 35.9% on tablet PCs. On rounds, 56% of doctors' tasks were performed in the corridors, 29% in patients' rooms, and 3% at the bedside. Doctors not on a ward round conducted 93.6% of tasks using stationary PCs, most often within the doctors' office. Nurses and doctors were observed performing workarounds, such as transcribing medication orders from the computer to paper. CONCLUSIONS: The choice of device was related to clinical role, nature of the clinical task, degree of mobility required, including where task completion occurs, and device design. Nurses' work, and clinical tasks performed by doctors during ward rounds, require highly mobile computer devices. Nurses and doctors on ward rounds showed a strong preference for generic COWs over all other devices. Tablet PCs were selected by doctors for only a small proportion of clinical tasks. Even when using mobile devices clinicians completed a very low proportion of observed tasks at the bedside. The design of the devices and ward space configurations place limitations on how and where devices are used and on the mobility of clinical work. In such circumstances, clinicians will initiate workarounds to compensate. In selecting hardware devices, consideration should be given to who will be using the devices, the nature of their work, and the physical layout of the ward. PMID- 19674961 TI - Ability to generate patient registries among practices with and without electronic health records. AB - BACKGROUND: The ability to generate registries of patients with particular clinical attributes, such as diagnoses or medications taken, is central to measuring and improving the quality of health care. However, it is not known how many providers have the ability to generate such registries. OBJECTIVES: To assess the proportion of physician practices that can construct registries of patients with specific diagnoses, laboratory results, or medications, and to determine the relationship between electronic health record (EHR) usage and the ability to perform registry functions. METHODS: We conducted a mail survey of a stratified random sample of physician practices in Massachusetts in the northeastern United States (N = 1884). The survey included questions about the physicians' ability to generate diagnosis, laboratory result, and medication registries; the presence of EHR; and usage of specific EHR features. RESULTS: The response rate was 71% (1345/1884). Overall, 79.8% of physician practices reported being able to generate registries of patients by diagnosis; 56.1% by laboratory result; and 55.8% by medication usage. In logistic regression analyses, adjusting for urban/rural location, practice size and ownership, teaching status, hospital affiliation, and specialty, physician practices with an EHR were more likely to be able to construct diagnosis registries (adjusted odds ratio [OR] 1.53, 95% confidence interval [CI] 1.25 - 1.86), laboratory registries (OR 1.42, 95% CI 1.22 - 1.66), and medication registries (OR 2.30, 95% CI 1.96 - 2.70). CONCLUSIONS: Many physician practices were able to generate registries, but this capability is far from universal. Adoption of EHRs appears to be a useful step toward this end, and practices with EHRs are considerably more likely to be able to carry out registry functions. Because practices need registries to perform broad-based quality improvement, they should consider adopting EHRs that have built-in registry functionality. PMID- 19674960 TI - Patients' attitudes toward electronic health information exchange: qualitative study. AB - BACKGROUND: In many countries, there has been substantial progress in establishing the electronic transmission of patients' health information between health care providers, but little is known about how best to engage patients in the process. OBJECTIVE: We explored patients' views about sharing of electronic health information and their preferences for learning about and participating in this process. METHODS: Patients in one Massachusetts community in the northeastern United States were recruited to participate in focus-group discussions. Prior to discussion, participants completed a written questionnaire that captured their reactions to draft educational materials and a consent form. The discussion moderator and two physicians analyzed the moderator's detailed notes from each session and participants' written comments, using an immersion crystallization approach. RESULTS: Three dominant themes emerged: (1) concerns about privacy and security, (2) the potential benefit to a person's health, and (3) the desire for more information about the consent process. On the pre discussion questionnaire, 55 out of 62 participants (88%) indicated that they would provide consent for their information to be shared electronically among their health care providers, given the materials they had reviewed. CONCLUSIONS: Patients are enthusiastic about electronic health information exchange, recognizing its capacity to improve the quality and safety of health care; however, they are also concerned about its potential to result in breached privacy and misuse of health data. As the exchange of electronic health information becomes more widespread, policy makers will need to ensure that patients have access to concise educational materials and opportunities to engage in conversations about the risks and benefits of participation. PMID- 19674962 TI - Test-retest reliability of web-based retrospective self-report of tobacco exposure and risk. AB - BACKGROUND: Retrospectively collected data about the development and maintenance of behaviors that impact health are a valuable source of information. Establishing the reliability of retrospective measures is a necessary step in determining the utility of that methodology and in studying behaviors in the context of risk and protective factors. OBJECTIVE: The goal of this study was to examine the reliability of self-report of a specific health-affecting behavior, tobacco use, and its associated risk and protective factors as examined with a Web-based questionnaire. METHODS: Core tobacco use and risk behavior questions in the Lifetime Tobacco Use Questionnaire-a closed, invitation-only, password controlled, Web-based instrument-were administered at a 2-month test-retest interval to a convenience sample of 1229 respondents aged 18 to 78 years. Tobacco use items, which covered cigarettes, cigars, smokeless tobacco, and pipe tobacco, included frequency of use, amount used, first use, and a pack-years calculation. Risk-related questions included family history of tobacco use, secondhand smoke exposure, alcohol use, and religiosity. RESULTS: Analyses of test-retest reliability indicated modest (.30 to .49), moderate (.50 to .69), or high (.70 to 1.00) reliability across nearly all questions, with minimal reliability differences in analyses by sex, age, and income grouping. Most measures of tobacco use history showed moderate to high reliability, particularly for age of first use, age of first weekly and first daily smoking, and age at first or only quit attempt. Some measures of family tobacco use history, secondhand smoke exposure, alcohol use, and religiosity also had high test-retest reliability. Reliability was modest for subjective response to first use. CONCLUSIONS: The findings reflect the stability of retrospective recall of tobacco use and risk factor self-report responses in a Web-questionnaire context. Questions that are designed and tested with psychometric scrutiny can yield reliable results in a Web setting. PMID- 19674963 TI - Concurrent quantification of proteome and phosphoproteome to reveal system-wide association of protein phosphorylation and gene expression. AB - Reversible phosphorylation of proteins is an important process modulating cellular activities from upstream, which mainly involves sequential phosphorylation of signaling molecules, to downstream where phosphorylation of transcription factors regulates gene expression. In this study, we combined quantitative labeling with multidimensional liquid chromatography-mass spectrometry to monitor the proteome and phosphoproteome changes in the initial period of adipocyte differentiation. The phosphorylation level of a specific protein may be regulated by a kinase or phosphatase without involvement of gene expression or as a phenomenon that accompanies the alteration of its gene expression. Concurrent quantification of phosphopeptides and non-phosphorylated peptides makes it possible to differentiate cellular phosphorylation changes at these two levels. Furthermore, on the system level, certain proteins were predicted as the targeted gene products regulated by identified transcription factors. Among them, several proteins showed significant expression changes along with the phosphorylation alteration of their transcription factors. This is to date the first work to concurrently quantify proteome and phosphoproteome changes during the initial period of adipocyte differentiation, providing an approach to reveal the system-wide association of protein phosphorylation and gene expression. PMID- 19674964 TI - Affinity enrichment and characterization of mucin core-1 type glycopeptides from bovine serum. AB - The lack of consensus sequence, common core structure, and universal endoglycosidase for the release of O-linked oligosaccharides makes O glycosylation more difficult to tackle than N-glycosylation. Structural elucidation by mass spectrometry is usually inconclusive as the CID spectra of most glycopeptides are dominated by carbohydrate-related fragments, preventing peptide identification. In addition, O-linked structures also undergo a gas-phase rearrangement reaction, which eliminates the sugar without leaving a telltale sign at its former attachment site. In the present study we report the enrichment and mass spectrometric analysis of proteins from bovine serum bearing Galbeta1 3GalNAcalpha (mucin core-1 type) structures and the analysis of O-linked glycopeptides utilizing electron transfer dissociation and high resolution, high mass accuracy precursor ion measurements. Electron transfer dissociation (ETD) analysis of intact glycopeptides provided sufficient information for the identification of several glycosylation sites. However, glycopeptides frequently feature precursor ions of low charge density (m/z > approximately 850) that will not undergo efficient ETD fragmentation. Exoglycosidase digestion was utilized to reduce the mass of the molecules while retaining their charge. ETD analysis of species modified by a single GalNAc at each site was significantly more successful in the characterization of multiply modified molecules. We report the unambiguous identification of 21 novel glycosylation sites. We also detail the limitations of the enrichment method as well as the ETD analysis. PMID- 19674965 TI - iTRAQ-based proteomics profiling reveals increased metabolic activity and cellular cross-talk in angiogenic compared with invasive glioblastoma phenotype. AB - Malignant gliomas (glioblastoma multiforme) have a poor prognosis with an average patient survival under current treatment regimens ranging between 12 and 14 months. The tumors are characterized by rapid cell growth, extensive neovascularization, and diffuse cellular infiltration of normal brain structures. We have developed a human glioblastoma xenograft model in nude rats that is characterized by a highly infiltrative non-angiogenic phenotype. Upon serial transplantation this phenotype will develop into a highly angiogenic tumor. Thus, we have developed an animal model where we are able to establish two characteristic tumor phenotypes that define human glioblastoma (i.e. diffuse infiltration and high neovascularization). Here we aimed at identifying potential biomarkers expressed by the non-angiogenic and the angiogenic phenotypes and elucidating the molecular pathways involved in the switch from invasive to angiogenic growth. Focusing on membrane-associated proteins, we profiled protein expression during the progression from an invasive to an angiogenic phenotype by analyzing serially transplanted glioma xenografts in rats. Applying isobaric peptide tagging chemistry (iTRAQ) combined with two-dimensional LC and MALDI TOF/TOF mass spectrometry, we were able to identify several thousand proteins in membrane-enriched fractions of which 1460 were extracted as quantifiable proteins (isoform- and species-specific and present in more than one sample). Known and novel candidate proteins were identified that characterize the switch from a non angiogenic to a highly angiogenic phenotype. The robustness of the data was corroborated by extensive bioinformatics analysis and by validation of selected proteins on tissue microarrays from xenograft and clinical gliomas. The data point to enhanced intercellular cross-talk and metabolic activity adopted by tumor cells in the angiogenic compared with the non-angiogenic phenotype. In conclusion, we describe molecular profiles that reflect the change from an invasive to an angiogenic brain tumor phenotype. The identified proteins could be further exploited as biomarkers or therapeutic targets for malignant gliomas. PMID- 19674966 TI - A community standard format for the representation of protein affinity reagents. AB - Protein affinity reagents (PARs), most commonly antibodies, are essential reagents for protein characterization in basic research, biotechnology, and diagnostics as well as the fastest growing class of therapeutics. Large numbers of PARs are available commercially; however, their quality is often uncertain. In addition, currently available PARs cover only a fraction of the human proteome, and their cost is prohibitive for proteome scale applications. This situation has triggered several initiatives involving large scale generation and validation of antibodies, for example the Swedish Human Protein Atlas and the German Antibody Factory. Antibodies targeting specific subproteomes are being pursued by members of Human Proteome Organisation (plasma and liver proteome projects) and the United States National Cancer Institute (cancer-associated antigens). ProteomeBinders, a European consortium, aims to set up a resource of consistently quality-controlled protein-binding reagents for the whole human proteome. An ultimate PAR database resource would allow consumers to visit one on-line warehouse and find all available affinity reagents from different providers together with documentation that facilitates easy comparison of their cost and quality. However, in contrast to, for example, nucleotide databases among which data are synchronized between the major data providers, current PAR producers, quality control centers, and commercial companies all use incompatible formats, hindering data exchange. Here we propose Proteomics Standards Initiative (PSI) PAR as a global community standard format for the representation and exchange of protein affinity reagent data. The PSI-PAR format is maintained by the Human Proteome Organisation PSI and was developed within the context of ProteomeBinders by building on a mature proteomics standard format, PSI-molecular interaction, which is a widely accepted and established community standard for molecular interaction data. Further information and documentation are available on the PSI PAR web site. PMID- 19674967 TI - Structural basis for parathyroid hormone-related protein binding to the parathyroid hormone receptor and design of conformation-selective peptides. AB - Parathyroid hormone (PTH) and PTH-related protein (PTHrP) are two related peptides that control calcium/phosphate homeostasis and bone development, respectively, through activation of the PTH/PTHrP receptor (PTH1R), a class B G protein-coupled receptor. Both peptides hold clinical interest for their capacities to stimulate bone formation. PTH and PTHrP display different selectivity for two distinct PTH1R conformations, but how their binding to the receptor differs is unclear. The high resolution crystal structure of PTHrP bound to the extracellular domain (ECD) of PTH1R reveals that PTHrP binds as an amphipathic alpha-helix to the same hydrophobic groove in the ECD as occupied by PTH, but in contrast to a straight, continuous PTH helix, the PTHrP helix is gently curved and C-terminally "unwound." The receptor accommodates the altered binding modes by shifting the side chain conformations of two residues within the binding groove: Leu-41 and Ile-115, the former acting as a rotamer toggle switch to accommodate PTH/PTHrP sequence divergence, and the latter adapting to the PTHrP curvature. Binding studies performed with PTH/PTHrP hybrid ligands having reciprocal exchanges of residues involved in different contacts confirmed functional consequences for the altered interactions and enabled the design of altered PTH and PTHrP peptides that adopt the ECD-binding mode of the opposite peptide. Hybrid peptides that bound the ECD poorly were selective for the G protein-coupled PTH1R conformation. These results establish a molecular model for better understanding of how two biologically distinct ligands can act through a single receptor and provide a template for designing better PTH/PTHrP therapeutics. PMID- 19674968 TI - The hepatitis C virus non-structural NS5A protein impairs both the innate and adaptive hepatic immune response in vivo. AB - The role of hepatitis C virus (HCV) protein non-structural (NS) 5A in HCV associated pathogenesis is still enigmatic. To investigate the in vivo role of NS5A for viral persistence and virus-associated pathogenesis a transgenic (Tg) mouse model was established. Mice with liver-targeted NS5A transgene expression were generated using the albumin promoter. Alterations in the hepatic immune response were determined by Western blot, infection by lymphocytic choriomeningitis virus (LCMV), and using transient NS3/4A Tg mice generated by hydrodynamic injection. Cytotoxic T lymphocyte (CTL) activity was investigated by the Cr-release assay. The stable NS5A Tg mice did not reveal signs of spontaneous liver disease. The intrahepatic immunity was disrupted in the NS5A Tg mice as determined by clearance of LCMV infection or transiently NS3/4A Tg hepatocytes in vivo. This impaired immunity was explained by a reduced induction of interferon beta, 2',5'-OAS, and PKR after LCMV infection and an impairment of the CTL mediated elimination of NS3-expressing hepatocytes. In conclusion, these data indicate that in the present transgenic mouse model, NS5A does not cause spontaneous liver disease. However, we discovered that NS5A could impair both the innate and the adaptive immune response to promote chronic HCV infection. PMID- 19674969 TI - Multiple Rieske proteins enable short- and long-term light adaptation of Synechocystis sp. PCC 6803. AB - In contrast to eukaryotes, most cyanobacteria contain several isoforms of the Rieske iron-sulfur protein, PetC, resulting in heterogeneity in the composition of the cytochrome b(6)f complexes. Of three isoforms in the mesophilic cyanobacterium Synechocystis PCC 6803, PetC1 is the major Rieske protein in the cytochrome b(6)f complex, whereas the physiological function of PetC2 and PetC3 is still uncertain. Comparison of wild type and various petC-deficient strains under selected light conditions revealed distinct functional differences: high light exposure of wild type cells resulted in a significantly enhanced petC2 transcript level, whereas a Delta petC1 mutant showed a low cytochrome b(6)f content, low electron flux, and a considerably increased accumulation of cytochrome-bd oxidase. In contrast to wild type and Delta petC1, Delta petC2 and Delta petC3 strains still grew fast under high-light conditions although all three Rieske proteins are required for maximal electron transport rates. Although the presence of PetC3 appears to be required for activation of the cyclic electron transport, state transitions were more effective in the absence of PetC2 and/or PetC3. In summary, our data suggest defined roles of the various PetC proteins in short- and long-term light adaptation. PMID- 19674970 TI - Critical role for GATA3 in mediating Tie2 expression and function in large vessel endothelial cells. AB - Endothelial phenotypes are highly regulated in space and time by both transcriptional and post-transcriptional mechanisms. There is increasing evidence that the GATA family of transcription factors function as signal transducers, coupling changes in the extracellular environment to changes in downstream target gene expression. Here we show that human primary endothelial cells derived from large blood vessels express GATA2, -3, and -6. Of these factors, GATA3 was expressed at the highest levels. In DNA microarrays of human umbilical vein endothelial cells (HUVEC), small interfering RNA-mediated knockdown of GATA3 resulted in reduced expression of genes associated with angiogenesis, including Tie2. At a functional level, GATA3 knockdown inhibited angiopoietin (Ang)-1 mediated but not vascular endothelial cell growth factor (VEGF)-mediated AKT signaling, cell migration, survival, and tube formation. In electrophoretic gel mobility shift assays and chromatin immunoprecipitation, GATA3 was shown to bind to regulatory regions within the 5'-untranslated region of the Tie2 gene. In co immunoprecipitation and co-transfection assays, GATA3 and the Ets transcription factor, ELF1, physically interacted and synergized to transactivate the Tie2 promoter. GATA3 knockdown blocked the ability of Ang-1 to attenuate vascular endothelial cell growth factor stimulation of vascular cell adhesion molecule-1 expression and monocytic cell adhesion. Moreover, exposure of human umbilical vein endothelial cells to tumor necrosis factor-alpha resulted in marked down regulation of GATA3 expression and reduction in Tie2 expression. Together, these findings suggest that GATA3 is indispensable for Ang-1-Tie2-mediated signaling in large vessel endothelial cells. PMID- 19674971 TI - Loss of calmodulin binding to Bax inhibitor-1 affects Pseudomonas-mediated hypersensitive response-associated cell death in Arabidopsis thaliana. AB - Bax inhibitor-1 (BI-1) is a cell death suppressor protein conserved across a variety of organisms. The Arabidopsis atbi1-1 plant is a mutant in which the C terminal 6 amino acids of the expressed BI-1 protein have been replaced by T-DNA insertion. This mutant BI-1 protein (AtBI-CM) produced in Escherichia coli can no longer bind to calmodulin. A promoter-reporter assay demonstrated compartmentalized expression of BI-1 during hypersensitive response, introduced by the inoculation of Pseudomonas syringae possessing the avrRTP2 gene, Pst(avrRPT2). In addition, both BI-1 knockdown plants and atbi1-1 showed increased sensitivity to Pst(avrRPT2)-induced cell death. The results indicated that the loss of calmodulin binding reduces the cell death suppressor activity of BI-1 in planta. PMID- 19674972 TI - Promoter regulation of the visinin-like subfamily of neuronal calcium sensor proteins by nuclear respiratory factor-1. AB - VILIP-1 (gene name VSNL1), a member of the neuronal Ca(2+) sensor protein family, acts as a tumor suppressor gene by inhibiting cell proliferation, adhesion, and invasiveness. VILIP-1 expression is down-regulated in several types of human cancer. In human non-small cell lung cancer, we found that down-regulation was due to epigenetic changes. Consequently, in this study we analyzed the VSNL1 promoter and its regulation. Serial truncation of the proximal 2-kb VSNL1 promoter (VP-1998) from its 5' terminus disclosed that the last 3' terminal 100 bp promoter fragment maintained similar promoter activity as compared with VP 1998 and therefore was referred to as VSNL1 minimal promoter. When the 5' terminal 50 bp were deleted from the minimal promoter, the activity was dramatically decreased, suggesting that the deleted 50 bp contained a potential cis-acting element crucial for promoter activity. Deletion and site-directed mutagenesis combined with in silico transcription factor binding analysis of VSNL1 promoter identified nuclear respiratory factor (NRF)-1/alpha-PAL as a major player in regulating VSNL1 minimal promoter activity. The function of NRF-1 was further confirmed using dominant-negative NRF-1 overexpression and NRF-1 small interfering RNA knockdown. Electrophoretic mobility shift assay and chromatin immunoprecipitation provided evidence for direct NRF-1 binding to the VSNL1 promoter. Methylation of the NRF-1-binding site was found to be able to regulate VSNL1 promoter activity. Our results further indicated that NRF-1 could be a regulatory factor for gene expression of the other visinin-like subfamily members including HPCAL4, HPCAL1, HPCA, and NCALD. PMID- 19674973 TI - Architectural nucleoporins Nup157/170 and Nup133 are structurally related and descend from a second ancestral element. AB - The nuclear pore complex (NPC) constitutes one of the largest protein assemblies in the eukaryotic cell and forms the exclusive gateway to the nucleus. The stable, approximately 15-20-MDa scaffold ring of the NPC is built from two multiprotein complexes arranged around a central 8-fold axis. Here we present crystal structures of two large architectural units, yNup170(979-1502) and hNup107(658-925) x hNup133(517-1156), each a constituent of one of the two multiprotein complexes. Conservation of domain arrangement and of tertiary structure suggests that Nup157/170 and Nup133 derived from a common ancestor. Together with the previously established ancestral coatomer element (ACE1), these two elements constitute the major alpha-helical building blocks of the NPC scaffold and define its branched, lattice-like architecture, similar to vesicle coats like COPII. We hypothesize that the extant NPC evolved early during eukaryotic evolution from a rudimentary structure composed of several identical copies of a few ancestral elements, later diversified and specified by gene duplication. PMID- 19674974 TI - Coordination between polymerase beta and FEN1 can modulate CAG repeat expansion. AB - The oxidized DNA base 8-oxoguanine (8-oxoG) is implicated in neuronal CAG repeat expansion associated with Huntington disease, yet it is unclear how such a DNA base lesion and its repair might cause the expansion. Here, we discovered size limited expansion of CAG repeats during repair of 8-oxoG in a wild-type mouse cell extract. This expansion was deficient in extracts from cells lacking pol beta and HMGB1. We demonstrate that expansion is mediated through pol beta multinucleotide gap-filling DNA synthesis during long-patch base excision repair. Unexpectedly, FEN1 promotes expansion by facilitating ligation of hairpins formed by strand slippage. This alternate role of FEN1 and the polymerase beta (pol beta) multinucleotide gap-filling synthesis is the result of uncoupling of the usual coordination between pol beta and FEN1. HMGB1 probably promotes expansion by stimulating APE1 and FEN1 in forming single strand breaks and ligatable nicks, respectively. This is the first report illustrating that disruption of pol beta and FEN1 coordination during long-patch BER results in CAG repeat expansion. PMID- 19674975 TI - Homologous recombination but not nucleotide excision repair plays a pivotal role in tolerance of DNA-protein cross-links in mammalian cells. AB - DNA-protein cross-links (DPCs) are unique among DNA lesions in their unusually bulky nature. The steric hindrance imposed by cross-linked proteins (CLPs) will hamper DNA transactions, such as replication and transcription, posing an enormous threat to cells. In bacteria, DPCs with small CLPs are eliminated by nucleotide excision repair (NER), whereas oversized DPCs are processed exclusively by RecBCD-dependent homologous recombination (HR). Here we have assessed the roles of NER and HR for DPCs in mammalian cells. We show that the upper size limit of CLPs amenable to mammalian NER is relatively small (8-10 kDa) so that NER cannot participate in the repair of chromosomal DPCs in mammalian cells. Moreover, CLPs are not polyubiquitinated and hence are not subjected to proteasomal degradation prior to NER. In contrast, HR constitutes the major pathway in tolerance of DPCs as judged from cell survival and RAD51 and gamma H2AX nuclear foci formation. Induction of DPCs results in the accumulation of DNA double strand breaks in HR-deficient but not HR-proficient cells, suggesting that fork breakage at the DPC site initiates HR and reactivates the stalled fork. DPCs activate both ATR and ATM damage response pathways, but there is a time lag between two responses. These results highlight the differential involvement of NER in the repair of DPCs in bacterial and mammalian cells and demonstrate the versatile and conserved role of HR in tolerance of DPCs among species. PMID- 19674976 TI - Mechanism of LDL binding and release probed by structure-based mutagenesis of the LDL receptor. AB - The LDL receptor (LDL-R) mediates cholesterol metabolism in humans by binding and internalizing cholesterol transported by LDL. Several different molecular mechanisms have been proposed for the binding of LDL to LDL-R at neutral plasma pH and for its release at acidic endosomal pH. The crystal structure of LDL-R at acidic pH shows that the receptor folds back on itself in a closed form, obscuring parts of the ligand binding domain with the epidermal growth factor (EGF)-precursor homology domain. We have used a structure-based site-directed mutagenesis approach to examine 12 residues in the extracellular domain of LDL-R for their effect on LDL binding and release. Our studies show that the interface between the ligand binding domain and the EGF-precursor homology domain seen at acidic pH buries residues mediating both LDL binding and release. Our results are consistent with an alternative model of LDL-R whereby multiple modules of the extracellular domain interact with LDL at neutral pH, concurrently positioning key residues so that at acidic pH the LDL-R:LDL interactions become unfavorable, triggering release. After LDL release, the closed form of LDL-R may target its return to the cell surface. PMID- 19674978 TI - A new subtype of frontotemporal lobar degeneration with FUS pathology. AB - Frontotemporal dementia (FTD) is a clinical syndrome with a heterogeneous molecular basis. The neuropathology associated with most FTD is characterized by abnormal cellular aggregates of either transactive response DNA-binding protein with Mr 43 kDa (TDP-43) or tau protein. However, we recently described a subgroup of FTD patients, representing around 10%, with an unusual clinical phenotype and pathology characterized by frontotemporal lobar degeneration with neuronal inclusions composed of an unidentified ubiquitinated protein (atypical FTLD-U; aFTLD-U). All cases were sporadic and had early-onset FTD with severe progressive behavioural and personality changes in the absence of aphasia or significant motor features. Mutations in the fused in sarcoma (FUS) gene have recently been identified as a cause of familial amyotrophic lateral sclerosis, with these cases reported to have abnormal cellular accumulations of FUS protein. Because of the recognized clinical, genetic and pathological overlap between FTD and amyotrophic lateral sclerosis, we investigated whether FUS might also be the pathological protein in aFTLD-U. In all our aFTLD-U cases (n = 15), FUS immunohistochemistry labelled all the neuronal inclusions and also demonstrated previously unrecognized glial pathology. Immunoblot analysis of protein extracted from post mortem aFTLD-U brain tissue demonstrated increased levels of insoluble FUS. No mutations in the FUS gene were identified in any of our patients. These findings suggest that FUS is the pathological protein in a significant subgroup of sporadic FTD and reinforce the concept that FTD and amyotrophic lateral sclerosis are closely related conditions. PMID- 19674981 TI - Cardiovascular flashlight. Percutaneous treatment of a giant right coronary artery pseudoaneurysm in Adamantiades-Behcet's syndrome. PMID- 19674979 TI - Overlap of disease susceptibility loci for rheumatoid arthritis and juvenile idiopathic arthritis. AB - BACKGROUND: Genome-wide association studies (GWAS) have been extremely successful in the search for susceptibility risk factors for complex genetic autoimmune diseases. As more studies are published, evidence is emerging of considerable overlap of loci between these diseases. In juvenile idiopathic arthritis (JIA), another complex genetic autoimmune disease, the strategy of using information from autoimmune disease GWAS or candidate gene studies to help in the search for novel JIA susceptibility loci has been successful, with confirmed association with two genes, PTPN22 and IL2RA. Rheumatoid arthritis (RA) is an autoimmune disease that shares similar clinical and pathological features with JIA and, therefore, recently identified confirmed RA susceptibility loci are also excellent JIA candidate loci. OBJECTIVE: To determine the overlap of disease susceptibility loci for RA and JIA. METHODS: /st> Fifteen single nucleotide polymorphisms (SNPs) at nine RA-associated loci were genotyped in Caucasian patients with JIA (n=1054) and controls (n=3531) and tested for association with JIA. Allele and genotype frequencies were compared between cases and controls using the genetic analysis software, PLINK. RESULTS: Two JIA susceptibility loci were identified, one of which was a novel JIA association (STAT4) and the second confirmed previously published associations of the TRAF1/C5 locus with JIA. Weak evidence of association of JIA with three additional loci (Chr6q23, KIF5A and PRKCQ) was also obtained, which warrants further investigation. CONCLUSION: All these loci are good candidates in view of the known pathogenesis of JIA, as genes within these regions (TRAF1, STAT4, TNFAIP3, PRKCQ) are known to be involved in T cell receptor signalling or activation pathways. PMID- 19674980 TI - Toll-like receptor 7 (TLR7) modulates anti-nucleosomal autoantibody isotype and renal complement deposition in mice exposed to syngeneic late apoptotic cells. AB - OBJECTIVES: The objectives of this study were to determine whether late apoptotic cell material directly induces autoantibodies characteristic of systemic lupus erythematosus (SLE) and to investigate the innate recognition pathways involved. METHODS: B6, B6.MyD88(-/-), B6.TLR7(-/-) and B6.TLR9(-/-) mice were subcutaneously injected with B6 syngeneic late apoptotic thymocytes (SLATs) without adjuvant on days 0, 10, 24 and 37. Sera were tested for IgG antibodies to histones and double-stranded DNA (dsDNA) by ELISA and Crithidia luciliae indirect immunofluorescence. IgG and C3 deposition in kidney glomeruli was assessed by immunostaining and fluorescence microscopy. RESULTS: SLAT injections induced anti dsDNA and anti-histone antibodies of the IgG1 and IgG2b isotypes in B6 but not MyD88(-/-) mice. TLR7(-/-) and TLR9(-/-) mice injected with SLATs produced delayed or slightly more robust responses, respectively. SLAT injections induced IgG deposits in renal glomeruli of B6, TLR7(-/-) and TLR9(-/-) mice that were absent in MyD88(-/-) mice. Unlike B6 and TLR9(-/-) animals, TLR7(-/-) mice failed to exhibit IgG colocalised glomerular C3 deposits and demonstrated autoantibodies of primarily the IgG2a isotype. CONCLUSIONS: Late apoptotic cell-induced anti histone and anti-dsDNA antibodies require MyD88 but not Toll-like receptor (TLR)9. Moreover, TLR7 promotes glomerular C3 deposition, possibly through a mechanism of altered antibody isotype switching. PMID- 19674982 TI - Aorto-left ventricular tunnel causing severe aortic regurgitation in adult man. PMID- 19674983 TI - The challenge of early systemic sclerosis for the EULAR Scleroderma Trial and Research group (EUSTAR) community. It is time to cut the Gordian knot and develop a prevention or rescue strategy. AB - Early diagnosis of systemic sclerosis (SSc) may allow the start of treatment that could slow disease progression. For this reason early diagnosis of the disease is of pivotal importance. However, the lack of diagnostic criteria and valid predictors significantly limit patient evaluation and the use of potentially effective drugs in the earliest phase of SSc. Early SSc may be suspected on the basis of Raynaud's phenomenon, puffy fingers, autoantibodies and SSc capillaroscopic pattern. In practice, the aim is to have criteria for the diagnosis of very early SSc. The criteria that are proposed are obviously provisional and need to be validated: (a) initially through a Delphi technique; (b) thereafter perhaps using already available datasets; but (c) of critical importance, through prospective studies. Only after prospective studies can these potential criteria be considered validated. The consensus on criteria for the classification of very early SSc might be part of the evolving EULAR/ACR project of reclassification of SSc. PMID- 19674984 TI - Mycobacterial arthritis of large joints. PMID- 19674985 TI - Rheumatoid arthritis after Chikungunya fever: a prospective follow-up study of 21 cases. PMID- 19674986 TI - Monozygotic twins with stiff person syndrome and autoimmune thyroiditis: rituximab inefficacy in a double-blind, randomised, placebo controlled crossover study. PMID- 19674987 TI - New rheumatoid arthritis genetic factor and C5 serum level. PMID- 19674988 TI - High incidence of cardiovascular events in patients with rheumatoid arthritis. PMID- 19674989 TI - The "tank top sign": a unique pattern of skin fibrosis seen in pansclerotic morphea. PMID- 19674990 TI - High prevalence of vertebral deformities in elderly patients with early rheumatoid arthritis. PMID- 19674991 TI - Opposite relationships between circulating Dkk-1 and cartilage breakdown in patients with rheumatoid arthritis and knee osteoarthritis. PMID- 19674992 TI - Nucleosome accumulation and reduction of C-reactive protein are associated with the generation of anti-nuclear antibodies in patients with rheumatoid arthritis treated with adalimumab, but not with etanercept. PMID- 19674993 TI - Significance of retropharyngeal lymph node dissection in hypopharyngeal cancer. AB - OBJECTIVE: We investigated the risk factors for metastasis to retropharyngeal lymph nodes (RPLNs) and the significance of dissection of RPLNs in hypopharyngeal cancer. Metastasis to the RPLNs is an important prognostic factor in head and neck cancer, especially in hypopharyngeal cancer. METHODS: Study subjects were 129 cases who received primary treatment at nine leading medical facilities in the field of head and neck cancer management in Japan. Focusing on RPLNs, we compared prognosis in RPLN-metastasis-positive, RPLN-metastasis-negative, RPLN dissected and RPLN-non-dissected cases. RESULTS: The 5-year survival rate for the entire study group was 41.1%. Metastasis to RPLNs occurred during the follow-up period in 13.2%. RPLN dissection was performed in 32 of the 129 cases at the time of primary treatment. In the RPLN-dissected group, the 5-year survival rate in the RPLN-metastasis-positive subgroup was 30.0%, whereas that in the RPLN metastasis-negative subgroup was 41.2%, showing no statistically significant difference. Among 17 cases having RPLN metastasis, 30.0% in the RPLN-dissected group (n = 10) survived for 5 years versus none in the RPLN-non-dissected group (n = 7). The rate of RPLN metastasis was higher in primary hypopharyngeal cancer of the posterior wall/post-cricoid area (PC/PW) compared with that of the piriform sinus (P = 0.020). CONCLUSIONS: We recommend RPLN dissection at the time primary of treatment of hypopharyngeal cancer, especially in cases with cancer at subsites PC/PW, as RPLN dissection is expected to improve prognosis. The primary subsites PC/PW are associated with a risk of RPLN metastasis. PMID- 19674994 TI - PSA doubling time as a predictive factor on repeat biopsy for detection of prostate cancer. AB - OBJECTIVE: Detection of prostate cancer needs a biopsy of the prostate. Suspecting cancer from an increase in prostate-specific antigen (PSA) has a high negative rate at an initial prostate biopsy. Cases with negative initial biopsy may be the candidates of subsequent biopsy. For lowering unnecessary repeat biopsy, the use of predictive factors before a repeat biopsy is applied for indication. METHODS: Seventy-seven cases with negative initial prostate biopsy received a repeat biopsy and factors for the detection of cancer were examined. RESULTS: PSA doubling time distinguished a part of cancer cases. Its sensitivity of 30, 50 and 70 months was 36.6%, 30.4% and 10%, respectively. Cancer case did not show PSA doubling time of >100 months in general. Values of PSA transition zone density, %Free/total PSA and PSA velocity were similar between cancer and no cancer cases. CONCLUSIONS: PSA doubling time was one of the predictive factors for the detection of prostate cancer and was valuable for avoiding unnecessary repeat biopsy in some cases. PMID- 19674995 TI - Flow cytometric detection of small cell lung cancer cells with aberrant CD45 expression in micrometastatic bone marrow. AB - A lot of hematologists are often faced with the difficulty of diagnosing bone marrow micrometastasis of carcinoma cells. We employed a new flow cytometric immunophenotyping by a combination of CD45 with three neuroendocrine markers: CD56, microtubule-associated protein-2 and synaptophysin, and successfully detected micrometastatic tumor cells in the bone marrow of a 61-year-old male patient with small cell lung cancer (SCLC), whose marrow smears never showed a distinct morphology of metastasis. It was noteworthy that these SCLC cells accompanied the aberrant expression of CD45, leukocyte common antigen known as a specific marker for hematolymphoid neoplasms, which was not detected in the tumor of primary lesion. We describe this rare case to arouse an attention that tumors of non-hematolymphoid origin can exhibit exceptional CD45-positvity in metastatic sites. PMID- 19674998 TI - Use of computational fluid dynamics to study forces exerted on prey by aquatic suction feeders. AB - Suction feeding is the most commonly used mechanism of prey capture among aquatic vertebrates. Most previous models of the fluid flow caused by suction feeders involve making several untested assumptions. In this paper, a Chimera overset grids approach is used to solve the governing equations of fluid dynamics in order to investigate the assumptions that prey do not interact with the flow and that the flow can be modelled as a one-dimensional flow. Results show that, for small prey, both neglecting the prey and considering prey interaction give similar calculated forces exerted on the prey. However, as the prey item increases in size toward the size of the gape, its effect on the flow becomes more pronounced. This in turn affects both the magnitude of the hydrodynamic forces imparted to the prey and the time when maximum force is delivered. Maximum force is delivered most quickly to intermediate sized prey, about one-third of mouth diameter, and most slowly to prey less than 7 per cent or greater than 67 per cent of mouth diameter. This suggests that the effect of prey size on the timing of suction forces may have substantial consequences for the feeding ecology of suction feeders that are known to prefer prey between 25 and 50 per cent of mouth diameter. Moreover, for a 15 cm fish with a 15 mm gape, assuming a radial one-dimensional flow field can result in underestimating the maximum force exerted on a 5 mm diameter spherical prey 1 gape distance from the mouth by up to 28.7 per cent. PMID- 19674997 TI - Suspension biomechanics of swimming microbes. AB - Micro-organisms play a vital role in many biological, medical and engineering phenomena. Some recent research efforts have demonstrated the importance of biomechanics in understanding certain aspects of micro-organism behaviours such as locomotion and collective motions of cells. In particular, spatio-temporal coherent structures found in a bacterial suspension have been the focus of many research studies over the last few years. Recent studies have shown that macroscopic properties of a suspension, such as rheology and diffusion, are strongly affected by meso-scale flow structures generated by swimming microbes. Since the meso-scale flow structures are strongly affected by the interactions between microbes, a bottom-up strategy, i.e. from a cellular level to a continuum suspension level, represents the natural approach to the study of a suspension of swimming microbes. In this paper, we first provide a summary of existing biomechanical research on interactions between a pair of swimming micro organisms, as a two-body interaction is the simplest many-body interaction. We show that interactions between two nearby swimming micro-organisms are described well by existing mathematical models. Then, collective motions formed by a group of swimming micro-organisms are discussed. We show that some collective motions of micro-organisms, such as coherent structures of bacterial suspensions, are satisfactorily explained by fluid dynamics. Lastly, we discuss how macroscopic suspension properties are changed by the microscopic characteristics of the cell suspension. The fundamental knowledge we present will be useful in obtaining a better understanding of the behaviour of micro-organisms. PMID- 19674999 TI - Probing aggressive motivation in a cichlid fish. AB - The duration of startles provides an inverse measure of motivation to resume the previous activity. Here, we use a novel method in which one convict cichlid fish (Amatitlania nigrofasciata) of a competing pair was startled independently of the opponent. Fish were given various opponents and the mean startle duration determined. This mean was negatively correlated with the mean use of highly escalated 'frontal activities' such as biting and frontal display, but not the less escalated lateral displays or tail beating. Thus the startle duration was a reliable surrogate measure of the most escalated components of aggressive interactions. That is, it provided a motivational probe for aggressiveness of individual fish. Fight motivation is often determined in terms of fight duration or physiological costs for losers, who reveal the costs they are prepared to pay. We discuss various potential advantages of the motivational probe over previous measures, particularly with respect to winners and losers and different times during the interactions. PMID- 19674996 TI - Quorum sensing and social networking in the microbial world. AB - For many years, bacterial cells were considered primarily as selfish individuals, but, in recent years, it has become evident that, far from operating in isolation, they coordinate collective behaviour in response to environmental challenges using sophisticated intercellular communication networks. Cell-to-cell communication between bacteria is mediated by small diffusible signal molecules that trigger changes in gene expression in response to fluctuations in population density. This process, generally referred to as quorum sensing (QS), controls diverse phenotypes in numerous Gram-positive and Gram-negative bacteria. Recent advances have revealed that bacteria are not limited to communication within their own species but are capable of 'listening in' and 'broadcasting to' unrelated species to intercept messages and coerce cohabitants into behavioural modifications, either for the good of the population or for the benefit of one species over another. It is also evident that QS is not limited to the bacterial kingdom. The study of two-way intercellular signalling networks between bacteria and both uni- and multicellular eukaryotes as well as between eukaryotes is just beginning to unveil a rich diversity of communication pathways. PMID- 19675000 TI - Fluctuating mate preferences in a marine fish. AB - According to theory, directional female choice for male sexual ornaments is expected to erode underlying genetic variation. Considerable attention, in this regard, has been given to understanding the ubiquity of heritable genetic variation in both female choice and male sexual traits. One intriguing possibility emerging from this work is that persistent genetic variation could be maintained, over time, by variation in female mate preferences. Here, we report the results of a four-year study showing significant year-to-year fluctuations in mate preferences in a small marine fish, the sand goby, Pomatoschistus minutus. Although the average size of mature fish varied across years, we were unable to find direct evidence linking this variation to differences in female preferences among years. Our results, nevertheless, underscore the importance of temporal fluctuations in female mate preferences, as these can have important consequences for understanding variation in sexual traits and the intensity of sexual selection. PMID- 19675001 TI - Predictive motor activation during action observation in human infants. AB - Certain regions of the human brain are activated both during action execution and action observation. This so-called 'mirror neuron system' has been proposed to enable an observer to understand an action through a process of internal motor simulation. Although there has been much speculation about the existence of such a system from early in life, to date there is little direct evidence that young infants recruit brain areas involved in action production during action observation. To address this question, we identified the individual frequency range in which sensorimotor alpha-band activity was attenuated in nine-month-old infants' electroencephalographs (EEGs) during elicited reaching for objects, and measured whether activity in this frequency range was also modulated by observing others' actions. We found that observing a grasping action resulted in motor activation in the infant brain, but that this activity began prior to observation of the action, once it could be anticipated. These results demonstrate not only that infants, like adults, display overlapping neural activity during execution and observation of actions, but that this activation, rather than being directly induced by the visual input, is driven by infants' understanding of a forthcoming action. These results provide support for theories implicating the motor system in action prediction. PMID- 19675002 TI - Polyandry enhances offspring survival in an infanticidal species. AB - The adaptive significance of polyandry is an intensely debated subject in sexual selection. For species with male infanticidal behaviour, it has been hypothesized that polyandry evolved as female counterstrategy to offspring loss: by mating with multiple males, females may conceal paternity and so prevent males from killing putative offspring. Here we present, to our knowledge, the first empirical test of this hypothesis in a combined laboratory and field study, and show that multiple mating seems to reduce the risk of infanticide in female bank voles Myodes glareolus. Our findings thus indicate that females of species with non-resource based mating systems, in which males provide nothing but sperm, but commit infanticide, can gain non-genetic fitness benefits from polyandry. PMID- 19675003 TI - Bitter taste perception in Neanderthals through the analysis of the TAS2R38 gene. AB - The bitter taste perception (associated with the ability or inability to taste phenylthiocarbamide) is mediated by the TAS2R38 gene. Most of the variation in this gene is explained by three common amino-acid polymorphisms at positions 49 (encoding proline or alanine), 262 (alanine or valine) and 296 (valine or isoleucine) that determine two common isoforms: proline-alanine-valine (PAV) and alanine-valine-isoleucine (AVI). PAV is the major taster haplotype (heterozygote and homozygote) and AVI is the major non-taster haplotype (homozygote). Amino acid 49 has the major effect on the distinction between tasters and non-tasters of all three variants. The sense of bitter taste protects us from ingesting toxic substances, present in some vegetables, that can affect the thyroid when ingested in large quantities. Balancing selection has been used to explain the current high non-taster frequency, by maintaining divergent TAS2R38 alleles in humans. We have amplified and sequenced the TAS2R38 amino acid 49 in the virtually uncontaminated Neanderthal sample of El Sidron 1253 and have determined that it was heterozygous. Thus, this Neanderthal was a taster individual, although probably slightly less than a PAV homozygote. This indicates that variation in bitter taste perception pre-dates the divergence of the lineages leading to Neanderthals and modern humans. PMID- 19675004 TI - Crowded locusts produce hatchlings vulnerable to fungal attack. AB - Transgenerational effects of parental experience on offspring immunity are well documented in the vertebrate literature (where antibodies play an obligatory role), but have only recently been described in invertebrates. We have assessed the impact of parental rearing density upon offspring disease resistance by challenging day-old locust hatchlings (Schistocerca gregaria) from either crowd- or solitary-reared parents with the fungal pathogen Metarhizium anisopliae var. acridum. When immersed in standardized conidia suspensions, hatchlings from gregarious parents suffered greater pathogen-induced mortality than hatchlings from solitary-reared parents. This observation contradicts the basic theory of positive density-dependent prophylaxis and demonstrates that crowding has a transgenerational influence upon locust disease resistance. PMID- 19675005 TI - Merging ancient and modern DNA: extinct seabird taxon rediscovered in the North Tasman Sea. AB - Ancient DNA has revolutionized the way in which evolutionary biologists research both extinct and extant taxa, from the inference of evolutionary history to the resolution of taxonomy. Here, we present, to our knowledge, the first study to report the rediscovery of an 'extinct' avian taxon, the Tasman booby (Sula tasmani), using classical palaeontological data combined with ancient and modern DNA data. Contrary to earlier work, we show an overlap in size between fossil and modern birds in the North Tasman Sea (classified currently as S. tasmani and Sula dactylatra fullagari, respectively). In addition, we show that Holocene fossil birds have mitochondrial control region sequences that are identical to those found in modern birds. These results indicate that the Tasman booby is not an extinct taxon: S. dactylatra fullagari O'Brien & Davies, 1990 is therefore a junior synonym of Sula tasmani van Tets, Meredith, Fullagar & Davidson, 1988 and all North Tasman Sea boobies should be known as S. d. tasmani. In addition to reporting the rediscovery of an extinct avian taxon, our study highlights the need for researchers to be cognizant of multidisciplinary approaches to understanding taxonomy and past biodiversity. PMID- 19675006 TI - Shifting latitudinal clines in avian body size correlate with global warming in Australian passerines. AB - Intraspecific latitudinal clines in the body size of terrestrial vertebrates, where members of the same species are larger at higher latitudes, are widely interpreted as evidence for natural selection and adaptation to local climate. These clines are predicted to shift in response to climate change. We used museum specimens to measure changes in the body size of eight passerine bird species from south-eastern Australia over approximately the last 100 years. Four species showed significant decreases in body size (1.8-3.6% of wing length) and a shift in latitudinal cline over that period, and a meta-analysis demonstrated a consistent trend across all eight species. Southern high-latitude populations now display the body sizes typical of more northern populations pre-1950, equivalent to a 7 degrees shift in latitude. Using ptilochronology, we found no evidence that these morphological changes were a plastic response to changes in nutrition, a likely non-genetic mechanism for the pattern observed. Our results demonstrate a generalized response by eight avian species to some major environmental change over the last 100 years or so, probably global warming. PMID- 19675007 TI - Inferring population histories using cultural data. AB - The question as to whether cultures evolve in a manner analogous to that of genetic evolution can be addressed by attempting to reconstruct population histories using cultural data. As others have argued, this can only succeed if cultures are isolated enough to maintain and pass on a central core of traditions that can be modified over time. In this study we used a set of cultural data (canoe design traits from Polynesia) to look for the kinds of patterns and relationships normally found in population genetic studies. After developing new techniques to accommodate the peculiarities of cultural data, we were able to infer an ancestral region (Fiji) and a sequence of cultural origins for these Polynesian societies. In addition, we found evidence of cultural exchange, migration and a serial founder effect. Results were stronger when analyses were based on functional traits (presumably subject to natural selection and convergence) rather than symbolic or stylistic traits (probably subject to cultural selection for rapid divergence). These patterns strongly suggest that cultural evolution, while clearly affected by cultural exchange, is also subject to some of the same processes and constraints as genetic evolution. PMID- 19675008 TI - Whether depositing fat or losing weight, fish maintain a balance. AB - In fish, the relative amount of tissues of different densities changes significantly over short periods throughout the year, depending on the availability of food, nutrition and their developmental status, such as sexual maturation. If a land-living animal accumulates fat it influences not only its general state of health, but also markedly increases its energy expenditure for locomotion owing to the force of gravity. On a body submerged in water, this force, which acts on the centre of gravity (COG), is counterbalanced by a lifting force that is negligible in air and which acts on the centre of buoyancy (COB). Any difference in the longitudinal positions of the two centres will therefore result in pitching moments that must be counteracted by body or fin movements. The displacement of the COG away from the COB is a result of tissues of different density (e.g. bones and fat) not being distributed homogeneously along the body axis. Moreover, the proportions of tissues of different densities change significantly with feeding status. It is still unknown whether these changes produce a displacement of the COG and thus affect the hydrostatic stability of fish. Analysis of computed tomography and magnetic resonance imaging images of Atlantic herring, Atlantic salmon and Atlantic mackerel reveals that the COG is fairly constant in each species, although we recorded major interspecies differences in the relative amount of fat, muscle and bone. We conclude that the distribution of different tissues along the body axis is very closely adjusted to the swimming mode of the fish by keeping the COG constant, independent of the body fat status, and that fish can cope with large variations in energy intake without jeopardizing their COG and thus their swimming performance. PMID- 19675009 TI - Evidence for regular ongoing introductions of mosquito disease vectors into the Galapagos Islands. AB - Wildlife on isolated oceanic islands is highly susceptible to the introduction of pathogens. The recent establishment in the Galapagos Islands of the mosquito Culex quinquefasciatus, a vector for diseases such as avian malaria and West Nile fever, is considered a serious risk factor for the archipelago's endemic fauna. Here we present evidence from the monitoring of aeroplanes and genetic analysis that C. quinquefasciatus is regularly introduced via aircraft into the Galapagos Archipelago. Genetic population structure and admixture analysis demonstrates that these mosquitoes breed with, and integrate successfully into, already established populations of C. quinquefasciatus in the Galapagos, and that there is ongoing movement of mosquitoes between islands. Tourist cruise boats and inter island boat services are the most likely mechanism for transporting Culex mosquitoes between islands. Such anthropogenic mosquito movements increase the risk of the introduction of mosquito-borne diseases novel to Galapagos and their subsequent widespread dissemination across the archipelago. Failure to implement and maintain measures to prevent the human-assisted transport of mosquitoes to and among the islands could have catastrophic consequences for the endemic wildlife of Galapagos. PMID- 19675010 TI - Seventy-five-million-year-old tropical tetra-like fish from Canada tracks Cretaceous global warming. AB - Newly discovered fossil fish material from the Cretaceous Dinosaur Park Formation of Alberta, Canada, documents the presence of a tropical fish in this northern area about 75 million years ago (Ma). The living relatives of this fossil fish, members of the Characiformes including the piranha and neon tetras, are restricted to tropical and subtropical regions, being limited in their distribution by colder temperatures. Although characiform fossils are known from Cretaceous through to Cenozoic deposits, none has been reported previously from North America. The modern distribution of characiforms in Mexico and southern Texas in the southernmost United States is believed to have been the result of a relatively recent colonization less than 12 Ma. The new Canadian fossils document the presence of these fish in North America in the Late Cretaceous, a time of significantly warmer global temperatures than now. Global cooling after this time apparently extirpated them from the northern areas and these fishes only survived in more southern climes. The lack of early Cenozoic characiform fossils in North America suggests that marine barriers prevented recolonization during warmer times, unlike in Europe where Eocene characiform fossils occur during times of global warmth. PMID- 19675011 TI - The ground level event 70 on December 13th, 2006 and related effective doses at aviation altitudes. AB - The 70th ground level event in the records of the Neutron Monitor network occurred on 13 December 2006 reaching a maximum count rate increase at the Oulu station of more than 90 % during the 5 min interval 3.05-3.10 UTC. Thereafter, count rates gradually decreased registering increases of a few per cent above the galactic cosmic ray background after a few hours. The primary proton spectrum during the first 6 h after the onset of the event is characterised in this work by fitting the energy and angular distribution by a power law in rigidity and a linear dependence in the pitch angle using a minimisation technique. The results were obtained by analysing the data from 28 Neutron Monitor stations. At very high northern and southern latitudes, the effective dose rates were estimated to reach values of 25-30 microSv h(-1) at atmospheric depth of 200 g cm(-2) during the maximum of the event. The increase in effective dose during north atlantic and polar flights was estimated to be in the order of 20 %. PMID- 19675012 TI - Antibiotic stewardship and consumption: findings from a pan-European hospital study. AB - OBJECTIVES: Much has been written about antibiotic stewardship although less is known about the structure and content of antibiotic policies at hospital level. As part of the European Commission Concerted Action Antibiotic Resistance Prevention And Control (ARPAC) Project, data on antibiotic stewardship were collated and relationships investigated by antibiotic consumption in European hospitals. METHODS: A questionnaire survey on antibiotic stewardship factors was completed by 170 hospitals from 32 European countries. Data on committees, antibiotic formularies and policies addressing empirical therapy and prophylaxis were collated. Data on antibiotic use, expressed as defined daily doses per 100 occupied bed-days (DDD/100 BD), were provided by 139 hospitals from 30 countries, and 124 hospitals provided both data sets. Six key indicator stewardship variables were analysed by European region, case mix and antibiotic consumption. RESULTS: Hospitals from Northern and Western Europe were more likely to convene antibiotic committees or drugs and therapeutic committees compared with those from Southern and South-Eastern Europe (P < 0.001). One-fifth of hospitals had neither an antibiotic committee nor a policy. Hospital antibiotic policies commonly included recommendations on individual drugs, drug choices, dosage, duration and route but were less likely to contain information on side effects and cost. There were no significant differences by median total (J01) antibiotic consumption, although other antibiotic subgroups differed by stewardship indicators. CONCLUSIONS: Policies and practices relating to antibiotic stewardship varied considerably across European hospitals. These data provide a benchmark for newer European strategies tackling antibiotic resistance. More work is required to achieve harmonization of recommended practice, particularly in hospitals from Southern Europe. PMID- 19675013 TI - Voluntary and mandatory surveillance for methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) bacteraemia in England. AB - Voluntary laboratory reporting of communicable disease, including bacteraemia, has been the mainstay of surveillance in England for >30 years. The impact of introducing a parallel mandatory reporting process for surveillance of Staphylococcus aureus bacteraemia [both methicillin susceptible (MSSA) and resistant (MRSA)] was assessed by national and regional comparison of MSSA and MRSA reports to the two surveillance systems. Introduction of mandatory reporting in 2001 demonstrated that the true number of cases was 40% higher than indicated by voluntary reporting (i.e. 60% case ascertainment by voluntary reporting). However by 2008 the difference in reporting of MRSA bacteraemia between the two systems dropped to 30%, with six of the nine health regions in England having improved their levels of voluntary reporting, although there was still under reporting from London, the South East and the North West. Improvements in voluntary surveillance contributed to increased ascertainment of bacteraemia due to S. aureus (both MRSA and MSSA). Decreasing trends for MRSA bacteraemia were evident in both surveillance systems, with a 56% decrease in the mandatory and a 53% decline in the voluntary systems, from 2004. In contrast there was little change in reported cases of MSSA during 2004-2006. However, in 2007, when MRSA bacteraemia case numbers decreased by 27%, MSSA bacteraemia case reports actually increased by 6%. Trends for MSSA bacteraemia can be assessed more accurately from voluntary than from mandatory surveillance at the present time because mandatory reporting of MSSA bacteraemia is incomplete, with only 133 of 170 (78%) Trusts reporting in all four quarters of a year. PMID- 19675014 TI - Antimicrobial resistance in the UK and Ireland. AB - After the dramatic expansion of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli and Klebsiella in the UK and Ireland from 2001 onwards, the situation appears to have stabilized, with similar ESBL prevalence detected in 2007 as in 2006. Equally dramatic, but more welcome, is the sharp reduction since 2005 in the number of methicillin-resistant Staphylococcus aureus (MRSA) bacteraemias in England, and the reduction in prevalence of MRSA as a proportion of all S. aureus bacteraemias. These two trends dominate the information from the major resistance surveillance schemes in the UK and Ireland, but a wealth of further detail is available from these rich information sources. Resistance rates vary between hospitals, between specialities within hospitals and between patients within specialities depending on their characteristics such as age. In addition, resistance varies over time as new genetic mechanisms appear in clinically relevant bacteria, and spread or retreat under the pressures of competition and changing patterns of antibiotic use. Up-to-date information is required, not only at a national level for research and policy purposes, but at a very local level to inform day-to-day clinical decisions. PMID- 19675015 TI - Infectious Disease Research Network. AB - The Infectious Disease Research Network (IDRN) has been in operation since 2001, with an overall aim to promote multidisciplinary collaborations and increase the capacity of multidisciplinary translational research in the UK. The Network has achieved this by utilizing several different forms of activity, including the running of research strategy workshops, organizing of protocol development groups and creation of a website containing several tools and resources. These resources include a researchers' database, funding and training bulletins, and hosting study webpages. The Network has >1500 members, and covers all regions in the UK. The IDRN website is www.idrn.org. Activity has encompassed several priority research areas, including antimicrobial resistance. PMID- 19675016 TI - Has the era of untreatable infections arrived? AB - Antibiotic resistance is a major public health concern, with fears expressed that we shortly will run out of antibiotics. In reality, the picture is more mixed, improving against some pathogens but worsening against others. Against methicillin-resistant Staphylococcus aureus (MRSA)--the highest profile pathogen- the range of treatment options is expanding, with daptomycin, linezolid and tigecycline all launched, and telavancin, ceftobiprole, ceftaroline and dalbavancin anticipated. There is a greater problem with enterococci, especially if, as in endocarditis, bactericidal activity is needed and the isolate has high level aminoglycoside resistance; nevertheless, daptomycin, telavancin and razupenem all offer cidal potential. Against Enterobacteriaceae, the rapid and disturbing spread of extended-spectrum beta-lactamases, AmpC enzymes and quinolone resistance is forcing increased reliance on carbapenems, with resistance to these slowly accumulating via the spread of metallo-, KPC and OXA 48 beta-lactamases. Future options overcoming some of these mechanisms include various novel beta-lactamase-inhibitor combinations, but none of these overcomes all the carbapenemase types now circulating. Multiresistance that includes carbapenems is much commoner in non-fermenters than in the Enterobacteriaceae, depending mostly on OXA carbapenemases in Acinetobacter baumannii and on combinations of chromosomal mutation in Pseudomonas aeruginosa. No agent in advanced development has much to offer here, though there is interest in modified, less-toxic, polymyxin derivatives and in the siderophore monobactam BAL30072, which has impressive activity against A. baumannii and members of the Burkholderia cepacia complex. A final and surprising problem is Neisseria gonorrhoeae, where each good oral agent has been eroded in turn and where there is now little in reserve behind the oral oxyimino cephalosporins, to which low level resistance is emerging. PMID- 19675017 TI - The changing epidemiology of resistance. AB - Antibiotic resistance is now a linked global problem. Dispersion of successful clones of multidrug resistant (MDR) bacteria is common, often via the movement of people. Local evolution of MDR bacteria is also important under the pressure of excessive antibiotic use, with horizontal gene transfer providing the means by which genes such as bla(CTX-M) spread amongst different bacterial species and strains. Beta-lactamase production is a common resistance mechanism in Gram negative bacteria, and the rapid dissemination of novel genes reflects their evolution under the selective pressure of antibiotic usage. Many Enterobacteriaceae now carry broad-spectrum beta-lactamases such as CTX-M, with particular genotypes associated with different geographical regions. The spread of these enzymes has compromised the clinical utility of a number of beta-lactam classes and with the spread of genes such as bla(KPC), carbapenems may be increasingly compromised in the future. High-level fluoroquinolone resistance (mainly caused by gyrA mutations) has also been shown to be associated with CTX-M and CMY-type enzymes, commonly due to co-carriage on conjugative plasmids of the gene for the aminoglycoside-inactivating enzyme AAC-6(1)-Ib-cr and qnr genes (which confer low-level resistance), allowing the easy selection of gyrA mutants in the host strain. Resistance in Gram-positive bacteria is also widely distributed and increasing, with the emergence of community-associated methicillin-resistant Staphylococcus aureus (MRSA) blurring the distinction between hospital and community strains. Antibiotic use and environmental factors all have a role in the emergence and spread of resistance. This article reviews some of the new mechanisms and recent trends in the global spread of MDR bacteria. PMID- 19675018 TI - New developments in HIV drug resistance. AB - Several new antiretroviral drugs have recently been licensed for use in HIV-1 infected patients. These include drugs in two new classes: an integrase inhibitor (raltegravir) and a CCR5 co-receptor antagonist (maraviroc). In addition, two new protease inhibitors, atazanavir and darunavir, which have activity against viruses resistant to other protease inhibitors, have come into clinical use. Finally etravirine, a novel non-nucleoside reverse transcriptase inhibitor (NNRTI) is being increasingly used in patients whose virus is resistant to the earlier NNRTIs. These clinical advances have required the development of novel assays and interpretation systems for detection of resistance to allow the laboratory monitoring of patients receiving these new therapies. PMID- 19675024 TI - Students' attitudes to the communications employed during an outbreak of meningococcal disease in a UK school: a qualitative study. AB - BACKGROUND: Following an outbreak of meningococcal disease in a school in the North West of England, the communication methods employed by the Health Protection Agency (HPA) were evaluated in order to explore ways of improving communication with the public. METHODS: Qualitative questionnaires were distributed to Year 12 (sixth form) students. The Framework approach was used to analyse the data, which were coded, and emergent themes identified. RESULTS: In the absence of clear communication from official sources, many participants suggested that circulating rumours caused confusion and anxiety in the student population. Rumours were spread through informal networks in person or through text and MSN messaging. It was generally perceived that accurate information in this period would have been useful to allay potentially unfounded anxiety. Most students surveyed reported that they were sufficiently aware of the situation prior to receiving official announcements. The information provided by the HPA through the school was generally perceived as being useful, but it came too late. CONCLUSION: In outbreak situations, rumours will spread rapidly in the absence of early communication, and this can be a significant cause of anxiety. The use of digital communication strategies should be considered, since they can seed dependable information that will disseminate rapidly through peer groups. PMID- 19675025 TI - Whole genome sequence of Desulfovibrio magneticus strain RS-1 revealed common gene clusters in magnetotactic bacteria. AB - Magnetotactic bacteria are ubiquitous microorganisms that synthesize intracellular magnetite particles (magnetosomes) by accumulating Fe ions from aquatic environments. Recent molecular studies, including comprehensive proteomic, transcriptomic, and genomic analyses, have considerably improved our hypotheses of the magnetosome-formation mechanism. However, most of these studies have been conducted using pure-cultured bacterial strains of alpha proteobacteria. Here, we report the whole-genome sequence of Desulfovibrio magneticus strain RS-1, the only isolate of magnetotactic microorganisms classified under delta-proteobacteria. Comparative genomics of the RS-1 and four alpha-proteobacterial strains revealed the presence of three separate gene regions (nuo and mamAB-like gene clusters, and gene region of a cryptic plasmid) conserved in all magnetotactic bacteria. The nuo gene cluster, encoding NADH dehydrogenase (complex I), was also common to the genomes of three iron-reducing bacteria exhibiting uncontrolled extracellular and/or intracellular magnetite synthesis. A cryptic plasmid, pDMC1, encodes three homologous genes that exhibit high similarities with those of other magnetotactic bacterial strains. In addition, the mamAB-like gene cluster, encoding the key components for magnetosome formation such as iron transport and magnetosome alignment, was conserved only in the genomes of magnetotactic bacteria as a similar genomic island-like structure. Our findings suggest the presence of core genetic components for magnetosome biosynthesis; these genes may have been acquired into the magnetotactic bacterial genomes by multiple gene-transfer events during proteobacterial evolution. PMID- 19675027 TI - Colles' fracture treated with non-bridging external fixation: a 1-year follow-up. AB - The results in 75 of 105 patients with Older type II/III (AO type A2.2, A3.1, A3.2) Colles' fractures, treated with non-bridging external fixation are presented. The mean age was 67.8 years, and all patients were followed prospectively for 12 months with radiological and functional assessment. No statistically significant loss of radial length, angulation or inclination was seen between the postoperative reduction and the 1-year follow-up examination. The clinical results after 1 year were 66 (88%) excellent/good, nine (12%) fair and 0 (0%) poor according to the modified Gartland and Werley score. Mean visual analogue scale pain score after 1 year was 0.8. In three patients (4%), re displacement of the fracture occurred and was treated with plating. Non-bridging external fixation offers a reliable method of maintaining radiological reduction of Older type II/III fractures of the distal radius and gives a good functional outcome after 1 year. PMID- 19675028 TI - Tendon entrapment in distal radius fractures. AB - We retrospectively defined the rate and clinical features of tendon entrapment in 693 consecutive patients with 701 distal radius fractures treated in a single hospital. Eight extensor tendons and one flexor tendon were entrapped. All fractures with extensor tendon entrapment were palmarly displaced (Smith type) or epiphyseal. Flexor tendon entrapment was seen in dorsally angulated (Colles type) epiphyseal fracture. The rate of tendon entrapment in acute distal radius fractures was 1.3%. Extensor tendon entrapment in palmarly displaced fractures is more common. PMID- 19675029 TI - Long-term results of low rotation humeral osteotomy in children with Erb's obstetric brachial plexus palsy. AB - Seventeen children with Erb's (C5/6 and C5/6/7 types) obstetric brachial plexus palsy who underwent low rotation humeral osteotomy to treat internal rotation contracture of the shoulder were recalled back to the clinic at a mean of 10 (range 8-14) years after surgery. Eight were male and nine female with mean age of 16 (range 13-20) years. The osteotomy procedure was done at a mean age of 6 (range 5-8) years. Preoperative, early postoperative, and late postoperative motor assessments were compared. There was no recurrence of the internal rotation posturing of the shoulder and there was maintenance of the improvements in elbow extension deficit and forearm rotation. The most surprising finding was a significant (P = 0.003) decrease in shoulder abduction on long-term follow-up (the mean shoulder abduction was 135 degrees , 146 degrees and 109 degrees measured pre-, early post- and late postoperatively, respectively). There was no correlation between changes in shoulder abduction and the radiological score of the shoulder. PMID- 19675030 TI - Histological staging and Dupuytren's disease recurrence or extension after surgical treatment: a retrospective study of 124 patients. AB - Dupuytren's disease has a high rate of recurrence after treatment. In this study we have assessed the usefulness of histological staging in the prediction of recurrence. We have also verified whether there is a correlation between histological staging and features of Dupuytren's diathesis. We studied 139 hands in 124 Caucasian patients treated between 1997 and 2004. There was a significant difference in the recurrence rate between the three histological types (P = 0.04). Histological staging was independent of features of Dupuytren's diathesis. This study confirms that histological staging is a reliable method for predicting recurrence. However, it should be used in association with clinical data to determine precisely the prognosis of patients suffering from Dupuytren's contracture. PMID- 19675031 TI - What is the significance of tendon suture purchase? AB - Repairs have been performed on porcine flexor tendons and subjected to tensile stress measurements to determine the effects and mechanism of core suture purchase (the length of the suture bite). Eighty-four pig trotter flexor profundus tendons were divided and repaired using four lengths of core suture purchase (1.33, 1, 0.66 and 0.33 cm) using a double modified Kessler repair (four strands, two knots) with a peripheral epitendinous suture. Tendon purchase was achieved by either bilateral equal purchase lengths or with one tendon purchase at a fixed depth of 1 cm. A separate group of tendons were incubated in blood for 24 hours to simulate the wound environment prior to testing. Tensile tests demonstrated a progressive increase of repair strength with purchase length. With the exception of the 0.33 cm group, video analysis demonstrated the mode of failure as suture failure and not due to suture pullout. Therefore, the increase in breaking strength cannot be attributed to a better grip of the tendon ends, but to the mechanical characteristics of the suture polymer. The tendency for the incubated tendons to fail more consistently by pullout rather than suture failure, particularly in the shorter purchase lengths, emphasises the importance of studying tendon purchase in vivo. The significance of ex vivo mechanical testing should be considered with caution. PMID- 19675032 TI - Mini open carpal tunnel release using Knifelight: evaluation of the safety and effectiveness of using a single wrist incision (cadaveric study). AB - This cadaveric study evaluates the margin of safety and technical efficacy of mini open carpal tunnel release performed using Knifelight (Stryker Instruments) through a transverse 1 cm wrist incision. A single investigator released 32 wrists in 17 cadavers. The wrists were then explored to assess the completeness of release and damage to vital structures including the superficial palmar arch, palmar cutaneous branch and recurrent branch of the median nerve. All the releases were complete and no injury to the median nerve and other structures were observed. The mean distance of the recurrent motor branch to the ligamentous divisions was 5.7 +/- 2.4 mm, superficial palmar arch was 8.7 +/- 3.1 mm and palmar cutaneous branch to the ligamentous division was 7.2 +/- 2.4 mm. The mean length of the transverse carpal ligament was 29.3 +/- 3.7 mm. Guyon's canal was preserved in all cases. PMID- 19675033 TI - Use of Wolfe graft for the treatment of mucous cysts. AB - Many surgical procedures have been described for the treatment of mucous cysts. We report a case series of a surgical technique that excises the cyst along with overlying skin and reconstructs the defect using a Wolfe graft harvested from the wrist crease. This technique can be applied to cysts in all locations, even those adjacent to the nail. The procedure has been performed on 51 mucous cysts with satisfactory results, a very low recurrence rate (4%) and negligible complications. PMID- 19675035 TI - Midcarpal instability after excision arthroplasty for scapho-trapezial-trapezoid (STT) arthritis. PMID- 19675034 TI - The effect of miniscalpel-needle versus steroid injection for trigger thumb release. AB - This study compared the result of percutaneous release using a miniscalpel-needle and steroid injection in 93 trigger thumbs in 83 patients. The patients were randomly assigned to either miniscalpel-needle percutaneous release (group A) or steroid injection (group B). Visual analogue pain scales and patients' satisfaction were evaluated at baseline, 1 and 12 months. One patient in group A and two patients in group B were lost to follow-up. Forty-four of the 46 trigger thumbs in group A and 12 of 47 trigger thumbs in group B had satisfactory results at 12 months. No digital nerve injury occurred in either group. Percutaneous release with a miniscalpel-needle had a higher success rate than steroid injection. PMID- 19675036 TI - Anconeus muscle flap for the treatment of soft tissue defects over the olecranon after total elbow arthroplasty. PMID- 19675037 TI - A simple technique for wire tensioning. PMID- 19675038 TI - The use of a trifurcation of the lateral antebrachial cutaneous nerve for digital nerve grafting. PMID- 19675039 TI - Wrist arthroscopy: beware the novice. PMID- 19675040 TI - Use of the A3 pulley as an interposition flap to cover periosteal defects. PMID- 19675041 TI - Intraosseous glomus tumour in a distal phalanx. PMID- 19675043 TI - Mallet finger as a complication of liquid nitrogen cryosurgery for verruca vulgaris. PMID- 19675042 TI - Arterial thrombosis in a replanted thumb due to factor V mutation and anti phospholipid antibodies. PMID- 19675044 TI - Closed traumatic A2 pulley rupture: rare mechanism of injury. PMID- 19675045 TI - Primary pyomyositis (bacterial myositis) of the pronator quadratus. PMID- 19675046 TI - A case of spontaneous wrist haematoma in Achenbach syndrome. PMID- 19675047 TI - Transcarpal migration of a broken Kirschner wire causing ulnar neurapraxia. PMID- 19675048 TI - Traumatic closed index extensor tendon rupture: a case report. PMID- 19675049 TI - Cutaneous leishmaniasis: a diagnosis of suspicion. PMID- 19675050 TI - Psychometric properties of the Patient Rated Wrist/Hand Evaluation - Dutch Language Version (PRWH/E-DLV). PMID- 19675051 TI - Re: Bulic K. Wassel type IV thumb duplication. A case of mistaken identity? J Hand Surg Eur. 2008, 33: 536-7. PMID- 19675052 TI - Re: Kalbermatten DF, Erba P, Mahay D, Wiberg M, Pierer G, Terenghi G. Schwann cell strip for peripheral nerve repair. J Hand Surg Eur. 2008, 33: 587-94. PMID- 19675053 TI - Re: Hansen TB, Vainorius D. High loosening rate of the Moje Acamo prosthesis for treating osteoarthritis of the trapeziometacarpal joint. J Hand Surg Eur. 2008, 33: 571-4. PMID- 19675054 TI - Re: Souer JS, Mudgal CS. Plate fixation in closed ipsilateral multiple metacarpal fractures. J Hand Surg Eur. 2008, 33: 740-4. PMID- 19675056 TI - Thyroid stimulating hormone cut-off to define subclinical hypothyroidism. PMID- 19675057 TI - The diagnosis and investigation of adrenal insufficiency in adults. AB - There is considerable variation in the methods used to diagnose and investigate adrenal insufficiency in clinical practice. These include a range of adrenocorticotropin (ACTH) stimulation and other dynamic testing protocols, serum cortisol cut-off values for diagnosis and tests used for differential diagnosis. With the introduction of modern cortisol and ACTH assays, the interpretation of tests used for diagnosis and differential diagnosis has become more complex and requires local validation. This review examines the basis of normal hypothalamic pituitary-adrenal axis function and adrenal insufficiency states based upon an evidence base accumulated over the past four decades. The role of the laboratory in the differential diagnosis and interpretation based upon assay methodology is discussed. The accurate identification of patients who may benefit from corticosteroid replacement in special settings such as critical illness is challenging and will be explored. PMID- 19675058 TI - Association of serum glycated albumin to haemoglobin A1C ratio with hepatic function tests in patients with chronic liver disease. AB - BACKGROUND: In patients with chronic liver disease (CLD), glycated haemoglobin (HbA(1C)) levels have been shown to be apparently lower than real values, whereas serum glycated albumin (GA) levels are apparently higher. The present study was aimed to examine whether both glycaemic indices are influenced by hepatic function. METHODS: Subjects consisted of 82 patients with CLD. Various indicators for hepatic function as well as HbA(1C) and GA were also measured. Estimated HbA(1C) values were calculated from the mean plasma glucose levels. Two hundred and two type 2 diabetic patients without CLD were studied as controls. RESULTS: Although GA was strongly correlated with HbA(1C) in patients with CLD as well as diabetic patients, GA levels in patients with CLD were relatively higher than those in diabetic patients. In patients with estimated HbA(1C) < or = 5.8%, GA levels significantly increased but HbA(1C) levels decreased as a function of decreasing hepaplastin test (HPT). The ratio of GA/HbA(1C) (G/H ratio) increased as a function of decreasing HPT. In patients with estimated HbA(1C) > 5.8%, in contrast, GA levels were independent of HPT levels. In the patients with CLD, GA and HbA(1C) were associated with mean plasma glucose levels and some indicators for hepatic function. The multivariate analysis revealed a significant association of G/H ratio with HPT, cholinesterase and direct bilirubin. The G/H ratio was not associated with the mean plasma glucose but with HPT and cholinesterase levels. CONCLUSIONS: The G/H ratio correlates with hepatic function but not with plasma glucose levels. Therefore, CLD should be suspected for diabetic patients with an elevated G/H ratio. PMID- 19675060 TI - Salt is getting under our skin. PMID- 19675059 TI - Prevalence of chronic kidney disease in patients with suspected sleep apnoea. AB - BACKGROUND: Little is known about the epidemiology of chronic kidney disease (CKD) in patients with suspected sleep apnoea (SA). METHODS: Glomerular filtration rate (eGFR) was calculated in consecutive patients referred for full night observed in-hospital polysomnography. SA was defined as the respiratory disturbance index (RDI) > 5. RESULTS: One hundred and fifty-eight patients were studied. The age (mean +/- SD) was 61.2 +/- 12.7 years, body mass index 29.5 +/- 5.9 kg/m(2) and eGFR 86.1 +/- 21.7 mL/min/1.73 m(2). SA was present in 133 patients (85%). The eGFR was 94.6 7 mL/min/1.73 m(2) in patients without SA and 84.5 7 mL/min/1.73 m(2) in patients with SA [mean difference (95% confidence interval) 10.0 (0.6-19.4) mL/min/1.73 m(2); P = 0.037]. Seventy patients had eGFR > or = 90 mL/min/1.73 m(2) (group 1), and 70 patients had between 60 and 89 mL/min/1.73 m(2) (group 2), and 18 patients had 30-59 mL/min/1.73 m(2) (CKD 3). Although the prevalence of SA did not differ among the groups (group 1: 80%; group 2: 86%; CKD 3: 94%), the number of central sleep apnoeas (CSA) per hour was 5.9 +/- 12.2 in CKD 3, six times greater compared to patients with eGFR > or = 60 mL/min/1.73 m(2) (1.0 +/- 2.1; P = 0.01). The prevalence of obstructive SA did not differ between the groups. After adjustment for age, gender, BMI, hypertension, diabetes mellitus and smoking status, CKD 3 (P = 0.0004) and New York Heart Association class > or =3 (P = 0.0001) remained predictive of CSA events per hour. CONCLUSIONS: eGFR is reduced in patients with SA, particularly in those with episodes of CSA. PMID- 19675061 TI - Comparison between creatinine-based equations for estimating total creatinine clearance in peritoneal dialysis: a multicentre study. AB - BACKGROUND: It is crucial to assess the adequacy of peritoneal dialysis (PD) because of its influence on patient outcome. Collecting dialysate and urine for 24 h can be rather troublesome, so a simple and inexpensive alternative method for rapidly evaluating adequacy in PD would be very useful. Our study aimed to assess the performance of 12 different creatinine (Cr)-based equations commonly used to estimate GFR in predicting total Cr clearance (totCrCL) in PD. METHODS: Four Italian dialysis centres enrolled 355 PD patients with 2916 fluid collections. To rank the equations, their accuracy (median absolute percentage error, MAPE), precision (root mean square error, RMSE), agreement (k statistics), sensitivity and specificity (area under ROC curves, AUC, where x = 1 - specificity and y = sensitivity) were calculated with reference to the measured totCrCL. RESULTS: The Gates, Virga and 4-MDRD equations showed the best global performance as concerns accuracy (MAPE = 14.1, 16.3, 15.9% respectively), precision (RMSE = 13.2, 13.3, 13.4), agreement (k = 0.425, 0.440, 0.375), sensitivity and specificity (AUC = 0.825, 0.826, 0.820), while the Cockcroft Gault formula revealed a rather poor reliability. CONCLUSIONS: Fluid collection remains the gold standard for assessing PD adequacy. Our study ascertained how 12 Cr-based equations performed in estimating totCrCL in PD patients with a view to enabling the most accurate and precise among them to be chosen for use in approximately assessing totCrCL. PMID- 19675062 TI - CMV findings in the gastrointestinal tract in kidney transplantation patients, patients with end-stage kidney disease and immunocompetent patients. AB - AIM: Cytomegalovirus (CMV) is the most common viral pathogen affecting organ transplant recipients. The objective was to determine to what extent CMV can be found in the gastrointestinal tract in kidney transplant recipients and to compare them with patients in dialysis and randomly chosen otherwise healthy patients who were referred for oesophagogastroduodenoscopy (OEGD) or colonoscopy. PATIENTS AND METHODS: Biopsies for CMV examinations were obtained from 130 oesophagogastroduodenoscopies and 54 colonoscopies performed on 82 kidney transplant recipients, 49 dialysis patients with chronic end-stage kidney disease and 53 immunocompetent patients because of clinical indications. CMV was demonstrated by immunohistochemistry, both in frozen sections using a monoclonal antibody against CMV-specific antigens (pp65 matrix protein) and in paraffin sections by means of a monoclonal antibody against the delayed early protein (p52). RESULTS: CMV-positive cells were found in the gastroduodenal mucosa in 46 (68%) out of 82 kidney transplant recipients, in 9 (31%) of 49 dialysis patients and in 15 (45%) of 53 immunocompetent patients, in the colorectal mucosa in 7 (50%), in 6 (30%) and in 9 (45%) of the patient groups, respectively. In the transplant recipient group, 4 patients had severe and 10 patients moderate CMV infection in the gastroduodenal mucosa. CMV disease was diagnosed in two patients with severe infection and in one patient with moderate infection. All dialysis and immunocompetent patients had only moderate or mild CMV involvement. CONCLUSION: It appears that CMV-positive cells were present in all groups studied, suggesting that CMV-infected cells alone are not sufficient to make the diagnosis of CMV disease in the transplanted host. Moreover, the clinical symptoms and the intensity of the histologic CMV findings did not correlate with the symptoms the patients were having. In kidney transplant recipients, it emerges that CMV is activated more easily in the upper rather than in the lower gastrointestinal tract. PMID- 19675063 TI - Renal involvement in AL amyloidosis: the facts, the promise and the hope. PMID- 19675065 TI - Kif5b is an essential forward trafficking motor for the Kv1.5 cardiac potassium channel. AB - We have investigated the role of the kinesin I isoform Kif5b in the trafficking of a cardiac voltage-gated potassium channel, Kv1.5. In Kv1.5-expressing HEK293 cells and H9c2 cardiomyoblasts, current densities were increased from control levels of 389 +/- 50.0 and 317 +/- 50.3 pA pF(1), respectively, to 614 +/- 74.3 and 580 +/- 90.9 pA pF(1) in cells overexpressing the Kif5b motor. Overexpression of the Kif5b motor increased Kv1.5 expression additively with several manipulations that reduce channel internalization, suggesting that it is involved in the delivery of the channel to the cell surface. In contrast, expression of a Kif5b dominant negative (Kif5bDN) construct increased Kv1.5 expression non additively with these manipulations. Thus, the dominant negative acts by indirectly inhibiting endocytosis. The increase in Kv1.5 currents induced by wild type Kif5b was dependent on Golgi function; a 6 h treatment with Brefeldin A reduced Kv1.5 currents to control levels in Kif5b-overexpressing cells but had little effect on the increase associated with Kif5bDN expression. Finally, expression of the Kif5bDN prior to induction of Kv1.5 in a tetracycline inducible system blocked surface expression of the channel in both HEK293 cells and H9c2 cardiomyoblasts. Thus, Kif5b is essential to anterograde trafficking of a cardiac voltage-gated potassium channel. PMID- 19675064 TI - Vagally mediated effects of glucagon-like peptide 1: in vitro and in vivo gastric actions. AB - Glucagon-like peptide-1 (GLP-1) is a neuropeptide released following meal ingestion that, among other effects, decreases gastric tone and motility. The central targets and mechanism of action of GLP-1 on gastric neurocircuits have not, however, been fully investigated. A high density of GLP-1 containing neurones and receptors are present in brainstem vagal circuits, suggesting that the gastroinhibition may be vagally mediated. We aimed to investigate: (1) the response of identified gastric-projecting neurones of the dorsal motor nucleus of the vagus (DMV) to GLP-1 and its analogues; (2) the effects of brainstem application of GLP-1 on gastric tone; and (3) the vagal pathway utilized by GLP-1 to induce gastroinhibition. We conducted our experiments using whole-cell recordings from identified gastric-projecting DMV neurones and microinjection in the dorsal vagal complex (DVC) of anaesthetized rats while monitoring gastric tone. Perfusion with GLP-1 induced a concentration-dependent excitation of a subpopulation of gastric-projecting DMV neurones. The GLP-1 effects were mimicked by exendin-4 and antagonized by exendin-9-39. In an anaesthetized rat preparation, application of exendin-4 to the DVC decreased gastric tone in a concentration-dependent manner. The gastroinhibitory effects of exendin-4 were unaffected by systemic pretreatment with the pro-motility muscarinic agonist bethanechol, but were abolished by systemic administration of the nitric oxide synthase (NOS) inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME), or by bilateral vagotomy. Our data indicate that GLP-1 activates selective receptors to excite DMV neurones mainly and that the gastroinhibition observed following application of GLP-1 in the DVC is due to the activation of an inhibitory non adrenergic, non-cholinergic input to the stomach. PMID- 19675066 TI - Asymmetric control of cycle period by the spinal locomotor rhythm generator in the adult cat. AB - During walking, a change in speed is accomplished by varying the duration of the stance phase, while the swing phase remains relatively invariant. To determine if this asymmetry in the control of locomotor cycles is an inherent property of the spinal central pattern generator (CPG), we recorded episodes of fictive locomotion in decerebrate cats with or without a complete spinal transection (acute or chronic). During fictive locomotion, stance and swing phases typically correspond to extension and flexion phases, respectively. The extension and flexion phases were determined by measuring the duration of extensor and flexor bursts, respectively. In the vast majority of locomotor episodes, cycle period varied more with the extension phase. This was found without phasic sensory feedback, supraspinal structures, pharmacology or sustained stimulation. We conclude that the control of walking speed is governed by an asymmetry within the organization of the spinal CPG, which can be modified by extraneous factors. PMID- 19675068 TI - Early biophysics of the NMDA receptor channel. PMID- 19675067 TI - Seeing the hand while reaching speeds up on-line responses to a sudden change in target position. AB - Goal-directed movements are executed under the permanent supervision of the central nervous system, which continuously processes sensory afferents and triggers on-line corrections if movement accuracy seems to be compromised. For arm reaching movements, visual information about the hand plays an important role in this supervision, notably improving reaching accuracy. Here, we tested whether visual feedback of the hand affects the latency of on-line responses to an external perturbation when reaching for a visual target. Two types of perturbation were used: visual perturbation consisted in changing the spatial location of the target and kinesthetic perturbation in applying a force step to the reaching arm. For both types of perturbation, the hand trajectory and the electromyographic (EMG) activity of shoulder muscles were analysed to assess whether visual feedback of the hand speeds up on-line corrections. Without visual feedback of the hand, on-line responses to visual perturbation exhibited the longest latency. This latency was reduced by about 10% when visual feedback of the hand was provided. On the other hand, the latency of on-line responses to kinesthetic perturbation was independent of the availability of visual feedback of the hand. In a control experiment, we tested the effect of visual feedback of the hand on visual and kinesthetic two-choice reaction times--for which coordinate transformation is not critical. Two-choice reaction times were never facilitated by visual feedback of the hand. Taken together, our results suggest that visual feedback of the hand speeds up on-line corrections when the position of the visual target with respect to the body must be re-computed during movement execution. This facilitation probably results from the possibility to map hand- and target-related information in a common visual reference frame. PMID- 19675069 TI - Dynamic changes in the direction of blood flow through the ductus arteriosus at birth. AB - Major cardiovascular changes occur at birth, including increased pulmonary blood flow (PBF) and closure of the ductus arteriosus (DA), which acts as a low resistance shunt between the fetal pulmonary and systemic circulations. Although the pressure gradient between these circulations reverses after birth, little is known about DA blood flow changes and whether reverse DA flow contributes to PBF after birth. Our aim was to describe the changes in PBF and DA flow before, during and after the onset of pulmonary ventilation at birth. Flow probes were implanted on the left pulmonary artery (LPA) and DA in preterm fetal sheep (n = 8) approximately 3 days before they were delivered and ventilated. Blood flow was measured in the LPA and DA, before and after umbilical cord occlusion (UCO) and for 2 h after ventilation onset. Following UCO, DA flow decreased from 534 +/- 57 ml min(1) to 237 +/- 29 ml min(1) which reflected a similar reduction in right ventricular output. Within 5 min of ventilation onset, PBF increased from 11 +/- 6 ml min(1) to 230 +/- 13 ml min(1) whereas DA flow decreased to 172 +/- 54 ml min(1); negative values indicate reverse DA flow (left-to-right shunting). Reverse flow through the DA contributed up to 50% of total PBF at 30 min and a decrease in this contribution accounted for 71 +/- 13% of the time-related decrease in PBF after birth. DA blood flow is very dynamic after birth and depends upon the pressure gradient between the pulmonary and systemic circulations. Following ventilation, reverse DA flow provided a significant contribution to total PBF after birth. PMID- 19675070 TI - Origin of complex behaviour of spatially discordant alternans in a transgenic rabbit model of type 2 long QT syndrome. AB - Enhanced dispersion of repolarization has been proposed as an important mechanism in long QT related arrhythmias. Dispersion can be dynamic and can be augmented with the occurrence of spatially out-of-phase action potential duration (APD) alternans (discordant alternans; DA). We investigated the role of tissue heterogeneity in generating DA using a novel transgenic rabbit model of type 2 long QT syndrome (LQT2). Littermate control (LMC) and LQT2 rabbit hearts (n = 5 for each) were retrogradely perfused and action potentials were mapped from the epicardial surface using di-4-ANEPPS and a high speed CMOS camera. Spatial dispersion (Delta APD and Delta slope of APD restitution) were both increased in LQT2 compared to LMC (Delta APD: 34 +/- 7 ms vs. 23 +/- 6 ms; Delta slope: 1.14 +/- 0.23 vs. 0.59 +/- 0.19). Onset of DA under a ramp stimulation protocol was seen at longer pacing cycle length (CL) in LQT2 compared to LMC hearts (206 +/- 24 ms vs. 156 +/- 5 ms). Nodal lines between regions with APD alternans out of phase from each other were correlated with conduction velocity (CV) alternation in LMC but not in LQT2 hearts. In LQT2 hearts, larger APD dispersion was associated with onset of DA at longer pacing CL. At shorter CLs, closer to ventricular fibrillation induction (VF), nodal lines in LQT2 (n = 2 out of 5) showed persistent complex beat-to-beat changes in nodal line formation of DA associated with competing contribution from CV restitution and tissue spatial heterogeneity, increasing vulnerability to conduction block. In conclusion, tissue heterogeneity plays a significant role in providing substrate for ventricular arrhythmia in LQT2 rabbits by facilitating DA onset and contributing to unstable nodal lines prone to reentry formation. PMID- 19675073 TI - Role of transitory carbon reserves during adjustment to climate variability and source-sink imbalances in oil palm (Elaeis guineensis). AB - Oil palm (Elaeis guineensis Jacq.) is a perennial, tropical, monocotyledonous plant characterized by simple architecture and low phenotypic plasticity, but marked by long development cycles of individual phytomers (a pair of one leaf and one inflorescence at its axil). Environmental effects on vegetative or reproductive sinks occur with various time lags depending on the process affected, causing source-sink imbalances. This study investigated how the two instantaneous sources of carbon assimilates, CO(2) assimilation and mobilization of transitory non-structural carbohydrate (NSC) reserves, may buffer such imbalances. An experiment was conducted in Indonesia during a 22-month period (from July 2006 to May 2008) at two contrasting locations (Kandista and Batu Mulia) using two treatments (control and complete fruit pruning treatment) in Kandista. Measurements included leaf gas exchange, dynamics of NSC reserves and dynamics of structural aboveground vegetative growth (SVG) and reproductive growth. Drought was estimated from a simulated fraction of transpirable soil water. The main sources of variation in source-sink relationships were (i) short term reductions in light-saturated leaf CO(2) assimilation rate (A(max)) during seasonal drought periods, particularly in Batu Mulia; (ii) rapid responses of SVG rate to drought; and (iii) marked lag periods between 16 and 29 months of environmental effects on the development of reproductive sinks. The resulting source-sink imbalances were buffered by fluctuations in NSC reserves in the stem, which mainly consisted of glucose and starch. Starch was the main buffer for sink variations, whereas glucose dynamics remained unexplained. Even under strong sink limitation, no negative feedback on A(max) was observed. In conclusion, the different lag periods for environmental effects on assimilate sources and sinks in oil palm are mainly buffered by NSC accumulation in the stem, which can attain 50% (dw:dw) in stem tops. The resulting dynamics of growth and production are complex because several dozen phytomers of different phenological ages develop at any given time and interact with a common pool of reserves. PMID- 19675071 TI - Downregulation of oxytocin and natriuretic peptides in diabetes: possible implications in cardiomyopathy. AB - Regular physical activity is beneficial in preventing the risk of cardiovascular complications of diabetes. Recent studies showed a cardioprotective role of oxytocin (OT) to induce natriuretic peptides (NPs) and nitric oxide (NO) release. It is not known if the diabetic state is associated with a reduced OT-NPs-NO system and if exercise training improves this system. To address this, we investigated the effects of treadmill running using the db/db mouse model of type 2 diabetes. Eight-week-old db/db mice were subjected to running 5 days per week for a period of 8 weeks. The lean db/+ littermates were used as controls. Sedentary db/db mice were obese and hyperglycaemic, and exercise training was not effective in reducing body weight and the hyperglycaemic state. Compared to control mice, db/db mice had lower heart weight and heart-to-body weight ratios. In these mice, this was associated with augmented cardiac apoptosis, cardiomyocyte enlargement and collagen deposits. In addition, db/db mice displayed significant downregulation in gene expression of OT (76%), OT receptors (65%), atrial NP (ANP; 43%), brain NP (BNP; 87%) and endothelial nitric oxide synthase (eNOS) (54%) in the heart (P < 0.05). Exercise training had no effect on expression of these genes which were stimulated in control mice. In response to exercise training, the significant increment of anti-apoptotic Bcl-2 gene expression was observed only in control mice (P < 0.05). In conclusion, downregulation of the OT-NPs-NO system occurs in the heart of the young db/db mouse. Exercise training was not effective in reversing the defect, suggesting impairment of this cardiac protective system in diabetes. PMID- 19675072 TI - High temperature does not alter fatigability in intact mouse skeletal muscle fibres. AB - Intense activation of skeletal muscle results in fatigue development, which involves impaired function of the muscle cells resulting in weaker and slower contractions. Intense muscle activity also results in increased heat production and muscle temperature may rise by up to 6 degrees C. Hyperthermia is associated with impaired exercise performance in vivo and recent studies have shown contractile dysfunction and premature fatigue development in easily fatigued muscle fibres stimulated at high temperatures and these defects were attributed to oxidative stress. Here we studied whether fatigue-resistant soleus fibres stimulated at increased temperature show premature fatigue development and whether increasing the level of oxidative stress accelerates fatigue development. Intact single fibres or small bundles of soleus fibres were fatigued by 600 ms tetani given at 2 s intervals at 37 degrees C and 43 degrees C, which is the highest temperature the muscle would experience in vivo. Tetanic force in the unfatigued state was not significantly different at the two temperatures. With 100 fatiguing tetani, force decreased by approximately 15% at both temperatures; the free cytosolic [Ca(2+)] (assessed with indo-1) showed a similar approximately 10% decrease at both temperatures. The oxidative stress during fatigue at 43 degrees C was increased by application of 10 microM hydrogen peroxide or tert butyl hydroperoxide and this did not cause premature fatigue development. In summary, fatigue-resistant muscle fibres do not display impaired contractility and fatigue resistance at the highest temperature that mammals, including humans, would experience in vivo. Thus, intrinsic defects in fatigue-resistant muscle fibres cannot explain the decreased physical performance at high temperatures. PMID- 19675074 TI - Wood CO(2) efflux and foliar respiration for Eucalyptus in Hawaii and Brazil. AB - We measured CO(2) efflux from wood for Eucalyptus in Hawaii for 7 years and compared these measurements with those on three- and four-and-a-half-year-old Eucalyptus in Brazil. In Hawaii, CO(2) efflux from wood per unit biomass declined approximately 10x from age two to age five, twice as much as the decline in tree growth. The CO(2) efflux from wood in Brazil was 8-10x lower than that for comparable Hawaii trees with similar growth rates. Growth and maintenance respiration coefficients calculated from Hawaii wood CO(2) efflux declined with tree age and size (the growth coefficient declined from 0.4 mol C efflux mol C( 1) wood growth at age one to 0.1 mol C efflux mol C(-1) wood growth at age six; the maintenance coefficient from 0.006 to 0.001 micromol C (mol C biomass)(-1) s( 1) at 20 degrees C over the same time period). These results suggest interference with CO(2) efflux through bark that decouples CO(2) efflux from respiration. We also compared the biomass fractions and wood CO(2) efflux for the aboveground woody parts for 3- and 7-year-old trees in Hawaii to estimate how focusing measurements near the ground might bias the stand-level estimates of wood CO(2) efflux. Three-year-old Eucalyptus in Hawaii had a higher proportion of branches < 0.5 cm in diameter and a lower proportion of stem biomass than did 7-year-old trees. Biomass-specific CO(2) efflux measured at 1.4 m extrapolated to the tree could bias tree level estimates by approximately 50%, assuming no refixation from bark photosynthesis. However, the bias did not differ for the two tree sizes. Foliar respiration was identical per unit nitrogen for comparable treatments in Brazil and Hawaii (4.2 micromol C mol N(-1) s(-1) at 20 degrees C). PMID- 19675075 TI - Aggressive inherited and sporadic medullary thyroid carcinomas display similar oncogenic pathways. AB - RET oncogene mutations are found in familial medullary thyroid carcinomas (MTC) and in one-third of sporadic cases. Oncogenic mechanisms involved in non-RET mutated sporadic MTC remain unclear. To study alterations associated with the development of both inherited and sporadic MTC, pangenomic DNA microarrays were used to analyze the transcriptome of 13 MTCs (four familial and nine sporadic). By using an ANOVA test, a list of 173 gene sequences with at least a twofold change expression was obtained. A subset of differentially expressed genes was controlled by real-time quantitative PCR and immunohistochemistry on a larger collection of MTCs. The expression pattern of those genes allowed us to distinguish two groups of sporadic tumors. The first group displays an expression profile similar to that expressed by inherited RET634 tumors. The second presents an expression profile close to that displayed by inherited RET918 tumors and includes tumors from patients with distant metastases. It is characterized by the overexpression of genes involved in proliferation and invasion (PTN, ESM1, and CEACAM6) or matrix remodeling (COL1A1, COL1A2, and FAP). Interestingly, RET918 tumors showed overexpression of the PTN gene, encoding pleiotrophin, a protein associated with metastasis. Using a MTC cell line, silencing of RET induced the inhibition of PTN gene expression. Overall, our results suggest that familial MTC and sporadic MTC could activate similar oncogenic pathways. PMID- 19675077 TI - A complication after a previous caesarean section. PMID- 19675076 TI - Estrogen receptor 1 mRNA is a prognostic factor in ovarian carcinoma: determination by kinetic PCR in formalin-fixed paraffin-embedded tissue. AB - Epidemiological and cell culture studies indicate that ovarian carcinoma growth is dependent on estrogen stimulation. However, possibly due to the lack of a reliable biomarker that helps to select patients according to prognostically relevant estrogen receptor (ER) levels, clinical trials using anti-estrogenic therapeutics in ovarian carcinoma have had inconsistent results. Therefore, we tested if ER expression analysis by a quantitative method might be useful in this regard in formalin-fixed paraffin-embedded (FFPE) tissue. In a study group of 114 primary ovarian carcinomas expression of estrogen receptor 1 (ESR1) mRNA was analyzed using a new method for RNA extraction from FFPE tissue that is based on magnetic beads, followed by kinetic PCR. The prognostic impact of ESR1 mRNA expression was investigated and compared to ERalpha protein expression as determined by immunohistochemistry. In univariate survival analysis the expression level of ESR1 mRNA was a significant positive prognostic factor for patient survival (hazard ratio (HR) 0.230 (confidence interval (CI) 0.102-0.516), P=0.002). ERalpha protein expression was correlated to ESR1 mRNA expression (P=0.0001); however, ERalpha protein expression did not provide statistically significant prognostic information. In multivariate analysis, ESR1 mRNA expression emerged as a prognostic factor, independent of stage, grade, residual tumor mass, age, and ERalpha protein expression (HR 0.227 (CI 0.078-0.656), P=0.006). Our results indicate that the determination of ESR1 levels by kinetic PCR may be superior to immunohistochemical methods in assessment of biologically relevant levels of ER expression in ovarian carcinoma, and is feasible in routinely used FFPE tissue. PMID- 19675078 TI - An unusual cause of chest pain. PMID- 19675080 TI - A multifaceted strategy for implementation of the Ottawa ankle rules in two emergency departments. AB - PROBLEM: Despite widespread acceptance of the Ottawa ankle rules for assessment of acute ankle injuries, their application varies considerably. DESIGN: Before and after study. BACKGROUND AND SETTING: Emergency departments of a tertiary teaching hospital and a community hospital in Australia. KEY MEASURES FOR IMPROVEMENT: Documentation of the Ottawa ankle rules, proportion of patients referred for radiography, proportion of radiographs showing a fracture. STRATEGIES FOR CHANGE: Education, a problem specific radiography request form, reminders, audit and feedback, and using radiographers as "gatekeepers." EFFECTS OF CHANGE: Documentation of the Ottawa ankle rules improved from 57.5% to 94.7% at the tertiary hospital, and 51.6% to 80.8% at the community hospital (P<0.001 for both). The proportion of patients undergoing radiography fell from 95.8% to 87.2% at the tertiary hospital, and from 91.4% to 78.9% at the community hospital (P<0.001 for both). The proportion of radiographs showing a fracture increased from 20.4% to 27.1% at the tertiary hospital (P=0.069), and 15.2% to 27.2% (P=0.002) at the community hospital. The missed fracture rate increased from 0% to 2.9% at the tertiary hospital and from 0% to 1.6% at the community hospital compared with baseline (P=0.783 and P=0.747). LESSONS LEARNT: Assessment of case note documentation has limitations. Clinician groups seem to differ in their capacity and willingness to change their practice. A multifaceted change strategy including a problem specific radiography request form can improve the selection of patients for radiography. PMID- 19675081 TI - Evidence based implementation of complex interventions. PMID- 19675082 TI - Diagnosis and management of neuropathic pain. PMID- 19675090 TI - Data show "alarming" rise in oral cancers among people in their 40s. PMID- 19675092 TI - Pharmacological management of gastrointestinal stromal tumours: an update on the role of sunitinib. AB - The efficacy and tolerability of the receptor tyrosine kinase inhibitor, sunitinib malate, have been demonstrated in phase I-III clinical trials of patients with imatinib-resistant or imatinib-intolerant gastrointestinal stromal tumours (GIST) as well as in a worldwide expanded-access study and in a continuous daily dosing (CDD) trial. Tumour genotype may have a significant influence on the activity of sunitinib in patients with imatinib-resistant GIST. Sunitinib activity was observed across different GIST genotypes and particularly in patients with wild-type and KIT exon 9 mutations (all relatively resistant to standard-dose imatinib) and in patients with secondary KIT exons 13 and 14 mutations. Adverse events with sunitinib were generally mild to moderate and easily managed by dose reduction, dose interruption or standard supportive measures. Treatment discontinuation can be avoided in most patients by close monitoring before and during treatment with appropriate adverse event management as necessary. The correlation between treatment exposure and clinical response is prompting the search for new approaches to treatment optimisation to ensure that patients derive maximum benefit from sunitinib therapy, including dose adjustments based on blood testing to ensure optimal drug exposure, and the use of the alternative CDD regimen to avoid treatment interruption. PMID- 19675093 TI - The strength of female sex as a prognostic factor in small-cell lung cancer: a pooled analysis of chemotherapy trials from the Manchester Lung Group and Medical Research Council Clinical Trials Unit. AB - BACKGROUND: Previous studies reported that women survive longer than men, but experience greater toxicity, when treated for small-cell lung cancer (SCLC). METHODS: Individual patient data from six randomized phase II/III chemotherapy trials, from the Manchester Lung Group and UK Medical Research Council, were pooled for analysis. End points included overall survival, response rate, toxicity, dose intensity (DI) and transfusion rates. RESULTS: Of 1707 patients analyzed, 44% were women. At baseline, women had poorer performance status (PS) (57% versus 67% Eastern Cooperative Oncology Group PS 0-1/Karnofsky PS 80-100, P = 0.0004) and more were of normal weight or underweight (57% versus 48%, P = 0.003), but fewer were anemic (25% versus 62%, P < 0.0001). Response rates between women and men were similar (77% versus 76%, P = 0.64). In univariate [hazard ratio (HR) 0.85, 95% confidence interval (CI) 0.76-0.96, P = 0.006] and multivariate (HR 0.88, 95% CI 0.79-0.99, P = 0.04) analyses, female sex predicted for longer survival. Women experienced more grade 3/4 emesis (18% versus 9%, P < 0.0001) and grade 3/4 mucositis (13% versus 8%, P = 0.005) than men. There were no differences in DI, infections, transfusions or treatment-related deaths. CONCLUSION: Data from >1700 patients in randomized SCLC chemotherapy trials confirm that women survive modestly longer than men but may experience greater toxicity. PMID- 19675094 TI - Neurexin 1 (NRXN1) deletions in schizophrenia. PMID- 19675091 TI - Metabolic glycoengineering: sialic acid and beyond. AB - This report provides a perspective on metabolic glycoengineering methodology developed over the past two decades that allows natural sialic acids to be replaced with chemical variants in living cells and animals. Examples are given demonstrating how this technology provides the glycoscientist with chemical tools that are beginning to reproduce Mother Nature's control over complex biological systems - such as the human brain - through subtle modifications in sialic acid chemistry. Several metabolic substrates (e.g., ManNAc, Neu5Ac, and CMP-Neu5Ac analogs) can be used to feed flux into the sialic acid biosynthetic pathway resulting in numerous - and sometime quite unexpected - biological repercussions upon nonnatural sialoside display in cellular glycans. Once on the cell surface, ketone-, azide-, thiol-, or alkyne-modified glycans can be transformed with numerous ligands via bioorthogonal chemoselective ligation reactions, greatly increasing the versatility and potential application of this technology. Recently, sialic acid glycoengineering methodology has been extended to other pathways with analog incorporation now possible in surface-displayed GalNAc and fucose residues as well as nucleocytoplasmic O-GlcNAc-modified proteins. Finally, recent efforts to increase the "druggability" of sugar analogs used in metabolic glycoengineering, which have resulted in unanticipated "scaffold-dependent" activities, are summarized. PMID- 19675096 TI - PerM: efficient mapping of short sequencing reads with periodic full sensitive spaced seeds. AB - MOTIVATION: The explosion of next-generation sequencing data has spawned the design of new algorithms and software tools to provide efficient mapping for different read lengths and sequencing technologies. In particular, ABI's sequencer (SOLiD system) poses a big computational challenge with its capacity to produce very large amounts of data, and its unique strategy of encoding sequence data into color signals. RESULTS: We present the mapping software, named PerM (Periodic Seed Mapping) that uses periodic spaced seeds to significantly improve mapping efficiency for large reference genomes when compared with state-of-the art programs. The data structure in PerM requires only 4.5 bytes per base to index the human genome, allowing entire genomes to be loaded to memory, while multiple processors simultaneously map reads to the reference. Weight maximized periodic seeds offer full sensitivity for up to three mismatches and high sensitivity for four and five mismatches while minimizing the number random hits per query, significantly speeding up the running time. Such sensitivity makes PerM a valuable mapping tool for SOLiD and Solexa reads. AVAILABILITY: http://code.google.com/p/perm/ PMID- 19675097 TI - The genomic structure of thermus bacteriophage {phi}IN93. AB - We have determined the complete nucleotide sequence of the phage IN93 is 19,604 bp long and contains 39 putative open reading frames. The functions for 20% of IN93 gene products are similar to those expressed by other known phages and bacteria, and include peptidase, lytic enzymes, integrase, repressor protein and replication protein. The structural proteins of the IN93 virion were identified through sodium dodecyl sulphate-polyacrylamide gel electrophoresis and found to have no similarity to those of other phages. We also determined the transcription initiation sites and classified four transcription units using the primer extension method. Three transcription units were transcribed in the same direction as part of the lytic cycle, while the remaining unit was transcribed in the opposite direction as part of the lysogenic cycle. PMID- 19675095 TI - Evolution of duplicated beta-globin genes and the structural basis of hemoglobin isoform differentiation in Mus. AB - The functional diversification of multigene families may be strongly influenced by mechanisms of concerted evolution such as interparalog gene conversion. The beta-globin gene family of house mice (genus Mus) represents an especially promising system for evaluating the effects of gene conversion on the functional divergence of duplicated genes. Whereas the majority of mammalian species possess tandemly duplicated copies of the adult beta-globin gene that are identical in sequence, natural populations of house mice are often polymorphic for distinct two-locus haplotypes that differ in levels of functional divergence between duplicated beta-globin genes, HBB-T1 and HBB-T2. Here, we use a phylogenetic approach to unravel the complex evolutionary history of the HBB-T1 and HBB-T2 paralogs in a taxonomically diverse set of species in the genus Mus. The main objectives of this study were 1) to reconstruct the evolutionary history of the different HBB haplotypes of house mice, 2) to assess the role of recombinational exchange between HBB-T1 and HBB-T2 in promoting concerted evolution, 3) to assess the role of recombinational exchange between HBB-T1 and HBB-T2 in creating chimeric genes, and 4) to assess the structural basis of hemoglobin isoform differentiation in species that possess distinct HBB paralogs. Results of our phylogenetic survey revealed that the HBB-T1 and HBB-T2 genes in different species of Mus exhibit the full range of evolutionary outcomes with respect to levels of interparalog divergence. At one end of the spectrum, the two identical HBB paralogs on the Hbb(s) haplotype (shared by Mus domesticus, Mus musculus, and Mus spretus) represent a classic example of concerted evolution. At the other end of the spectrum, the two distinct HBB paralogs on the Hbb(d), Hbb(p), Hbb(w1), and Hbb(w2) haplotypes (shared by multiple species in the subgenus Mus) show no trace of gene conversion and are distinguished by a number of functionally important amino acid substitutions. Because the possession of distinct HBB paralogs expands the repertoire of functionally distinct hemoglobin isoforms that can be synthesized during fetal development and postnatal life, variation in the level of functional divergence between HBB-T1 and HBB-T2 may underlie important physiological variation within and among species. PMID- 19675098 TI - PERIOD1 is an anti-apoptotic factor in human pancreatic and hepatic cancer cells. AB - PERIOD1 (PER1) is a clock gene. We examined the effect of knockdown of PER1 on apoptosis in pancreatic cancer (MIA PaCa-2 and PANC-1) and hepatocellular carcinoma (HepG2) cells. Transfection of siRNA against PER1 into these cells increased the cleaved forms of caspases and poly-ADP-ribose-polymerase and induced apoptosis in all three cell lines. In the two pancreatic cancer cell lines, PER1 knockdown resulted in upregulation of Bax and downregulation of Bcl 2. Expression of p53 was not altered in the two pancreatic cancer cell lines containing mutated p53, but was upregulated in the HepG2 cells containing wild type p53. Cell proliferation of MIA PaCa-2 and HepG2 was inhibited by PER1 knockdown. We also examined, by immunohistochemical staining, the expression of PER1 in pancreatic cancer tissue and found that PER1 was strongly expressed in pancreatic cancer cells. These results indicate that PER1 acts as an anti apoptotic factor in pancreatic cancer cells. PMID- 19675100 TI - Amyloid formation by the model protein muscle acylphosphatase is accelerated by heparin and heparan sulphate through a scaffolding-based mechanism. AB - Amyloid formation is the hallmark of many diseases. The propensity of a protein to aggregate depends on a number of biological factors like the presence of sulphated polysaccharides termed as glycosaminoglycans (GAGs). Here we assessed whether the polymeric nature of GAGs is responsible for their protein aggregation promoting effect. We studied the effect of different monosaccharide derivatives, featuring the main characteristics of heparin and heparan sulphate (HS) building blocks, on the aggregation kinetics of human muscle acylphosphatase (mAcP), a useful model protein for these studies. We observed that while heparin and HS changed the mAcP aggregation kinetic profile, the monosaccharide derivatives had no effect, whatever their concentration could be and both when they are studied separately or in combination. In contrast, heparin fragments with six or more monosaccharides reproduced the effects of HS and in part those of heparin. We conclude that the effect of heparin and HS on protein aggregation arises from the clustering and regular distribution of their composing units on a polymeric structure. We propose a model in which heparin and HS promote mAcP aggregation through a scaffolding-based mechanism, in which the regularly spaced sulphate moieties of the polymer interact with mAcP molecules increasing their local concentration and facilitating their orientation. PMID- 19675101 TI - Flaxseed oil supplementation increases plasma F1-phytoprostanes in healthy men. AB - Supplementation with eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) has been reported to reduce lipid peroxidation products formed from arachidonic acid (F(2)-isoprostanes) in healthy humans, as well as in those under oxidative stress. alpha-Linolenic acid (ALA) is a precursor to EPA and DHA; however, its conversion in humans is thought to be inefficient. ALA can also undergo free radical oxidation, forming compounds known as F(1)-phytoprostanes, which are found in all plants and are in high concentrations in plant pollens. In this study, we examined the effect of ALA supplementation on plasma and urine F(1) phytoprostane and F(2)-isoprostane concentrations in men. Thirty-six nonsmoking men, aged 20-65 y, were recruited from the general population and randomly allocated to consume 9 g/d of either flaxseed oil (62% ALA, 5.4 g/d) or olive oil (placebo) for 4 wk in a parallel design. At baseline and after 4 wk of supplementation, blood samples and a 24-h urine sample were collected for measurement of plasma and urinary F(1)-phytoprostanes and F(2)-isoprostanes and plasma fatty acids. Compared with the olive oil group, plasma phospholipid ALA was greater (P < 0.0001), as were F(1)-phytoprostanes in plasma (P = 0.049) and urine (P = 0.06) in the flaxseed oil group after 4 wk supplementation. Flaxseed oil did not affect plasma or urinary F(2)-isoprostanes. The greater plasma F(1) phytoprostane concentration in the flaxseed oil group most likely resulted from the increased plasma concentration of the ALA substrate and/or the F(1) phytoprostane content of the flaxseed oil. Future studies are needed to determine the physiological importance of increased plasma and urine F(1)-phytoprostanes and their relevance to heart disease prevention. PMID- 19675099 TI - Ro52-mediated monoubiquitination of IKK{beta} down-regulates NF-{kappa}B signalling. AB - Upon activation, NF-kappaB translocates into the nucleus and initiates biological events. This NF-kappaB signalling is mainly regulated by the protein kinase IKKbeta. Early in this signalling pathway, IKKbeta is phosphorylated for activation by several factors, such as pro-inflammatory cytokines and the Tax oncoprotein of HTLV-1. In cells infected by HTLV-1, IKKbeta is persistently phosphorylated and conjugated with monoubiquitin due to Tax expression. Although this Tax-induced monoubiquitination appears to be an important regulation system for IKKbeta, how the monoubiquitination occurs is unknown and its role in NF kappaB signalling is still unclear. Here, we show that an E3-ubiquitin ligase Ro52 interacts weakly with wild-type IKKbeta but strongly with a phosphomimetic mutant IKKbeta to conjugate monoubiquitin in cooperation with an E2-ubiquitin conjugating enzyme UbcH5B. These results suggest that the Tax-induced phosphorylation of IKKbeta causes an interaction with Ro52 for the subsequent monoubiquitination. NF-kappaB reporter assays have shown that the IKKbeta activity is suppressed by wild-type Ro52, but not by its inactive mutant. In addition, monoubiquitin fusion of IKKbeta reduced its activity for NF-kappaB signalling. We also found that Ro52 dramatically reduces the level of Tax. These results suggest that Ro52 down-regulates Tax-induced NF-kappaB signalling by monoubiquitinating IKKbeta and by reducing the level of Tax. PMID- 19675102 TI - Diet quality is associated with better cognitive test performance among aging men and women. AB - Most studies of association between diet and cognition among the elderly focus on the role of single nutrients or foods and ignore the complexity of dietary patterns and total diet quality. We prospectively examined associations between an index of diet quality and cognitive function and decline among elderly men and women of the Cache County Study on Memory and Aging in Utah. In 1995, 3634 resident men and women > or =65 y of age completed a baseline survey that included a 142-item FFQ. Cognition was assessed using an adapted version of the Modified Mini-Mental State Examination (3MS) at baseline and 3 subsequent interviews spanning approximately 11 y. A recommended food score (RFS) and non RFS were computed by summing the number of recommended foods (n = 57) and nonrecommended foods (n = 23) regularly consumed. Multivariable-mixed models were used to estimate associations between the RFS and non-RFS and average 3MS score over time. Those in the highest quartile of RFS scored 1.80 points higher on the baseline 3MS test than did those in the lowest quartile of RFS (P < 0.001). This effect was strengthened over 11 y of follow-up. Those with the highest RFS declined by 3.41 points over 11 y compared with the 5.2-point decline experienced by those with the lowest RFS (P = 0.0013). The non-RFS was not associated with cognitive scores. Consuming a diverse diet that includes a variety of recommended foods may help to attenuate age-related cognitive decline among the elderly. PMID- 19675104 TI - Multiple micronutrients including iron are not more effective than iron alone for improving hemoglobin and iron status of Malian school children. AB - Iron deficiency and anemia remain among the most important global public health problems facing school children. Helminth infections often peak at school age and aggravate nutritional risks. We conducted a 12-wk randomized controlled trial in 406 Malian anemic schoolchildren infected with Schistosoma hematobium to examine the effects of 2 doses of praziquantel (P) (40 mg/kg body weight), P + 60 mg/d iron (Fe), and/or a multiple micronutrient supplement (MM) that included 18 mg/d Fe. Supplements were administered to the children each school day (5 d/wk) throughout the study. Changes in hemoglobin (Hb), serum ferritin (SF), and serum transferrin receptors (s-TfR) were followed. We also examined interactions between Fe and MM supplements on Hb and SF concentrations and malaria incidence. The effects of Fe on Hb and SF concentrations were greater than the effects of P alone and MM with or without added Fe at 6 and 12 wk (P < 0.001). In all groups, s-TfR decreased at 6 and 12 wk compared with baseline. The decrease was most pronounced in the P + Fe group compared with the other 3 groups at wk 6 (P = 0.05). Fe and MM interacted negatively at wk 6 and 12 to affect Hb (beta = -0.43, 95% CI = -0.77, -0.09; P = 0.01 and beta = -0.47, 95% CI = -0.83, -0.11; P = 0.01, respectively) and SF (beta = -0.42, 95% CI = -25.60, 12.31; P < 0.001, and beta = -0.37, 95% CI = -0.63, -0.12; P = 0.004, respectively). Malaria incidence was higher in the groups treated with added Fe (relative risk: 1.66; 95% CI: 0.75, 3.67). In this context, MM with added iron were not more effective than Fe without MM. Fe supplementation of schoolchildren with 60 mg/d for anemia control should be considered carefully. PMID- 19675103 TI - Bacillus polyfermenticus ameliorates colonic inflammation by promoting cytoprotective effects in colitic mice. AB - Although human consumption of Bacillus polyfermenticus provides several health benefits, the probiotic effect of this bacterium against colonic inflammation has not yet, to our knowledge, been studied. Therefore, we induced colitis in mice by oral or intrarectal administration of dextran sodium sulfate (DSS) or trinitrobenzenosulfonic acid (TNBS), respectively, and investigated the effect of B. polyfermenticus on colitis. We found that mice treated with DSS or TNBS along with B. polyfermenticus had reduced mortality and severity of colitis (weight loss, diarrhea, and mucosal damages) than mice treated with DSS or TNBS alone. B. polyfermenticus also reduced the expression of inflammatory molecules, including chemokine (C-X-C motif) ligand 1, intercellular adhesion molecule, and tumor necrosis factor-alpha, but enhanced the expression of the antiinflammatory cytokine interleukin-10 in the inflamed mouse colon. Moreover, B. polyfermenticus suppressed apoptosis both in vivo in inflamed colonic mucosa and in vitro in colonic epithelial cells stimulated with apoptosis-inducing agents (FasL or Clostridium difficile Toxin A) when the apoptotic response was determined by a terminal deoxynucleotidyl transferase dUTP nick end labeling assay and cleavage of poly(ADP-ribose) polymerase or caspase-3, respectively. Treating colonic epithelial cells with B. polyfermenticus-conditioned medium (BPCM) enhanced cell proliferation and induced the phosphoinositide 3-kinases/Akt signaling pathway, suggesting that this bacterium can promote epithelial cell proliferation. BPCM also promoted the migration of colonic epithelial cells. These data suggest that B. polyfermenticus ameliorates colonic inflammation by suppressing apoptosis and promoting epithelial cell proliferation and migration. PMID- 19675105 TI - Dual-process action of exercise on appetite control: increase in orexigenic drive but improvement in meal-induced satiety. AB - BACKGROUND: Exercise could contribute to weight loss by altering the sensitivity of the appetite regulatory system. OBJECTIVE: The aim of this study was to assess the effects of 12 wk of mandatory exercise on appetite control. DESIGN: Fifty eight overweight and obese men and women [mean (+/-SD) body mass index (in kg/m(2)) = 31.8 +/- 4.5, age = 39.6 +/- 9.8 y, and maximal oxygen intake = 29.1 +/- 5.7 mL . kg(-1) . min(-1)] completed 12 wk of supervised exercise in the laboratory. The exercise sessions were designed to expend 2500 kcal/wk. Subjective appetite sensations and the satiating efficiency of a fixed breakfast were compared at baseline (week 0) and at week 12. An Electronic Appetite Rating System was used to measure subjective appetite sensations immediately before and after the fixed breakfast in the immediate postprandial period and across the whole day. The satiety quotient of the breakfast was determined by calculating the change in appetite scores relative to the breakfast's energy content. RESULTS: Despite large variability, there was a significant reduction in mean body weight (3.2 +/- 3.6 kg), fat mass (3.2 +/- 2.2 kg), and waist circumference (5.0 +/- 3.2 cm) after 12 wk. The analysis showed that a reduction in body weight and body composition was accompanied by an increase in fasting hunger and in average hunger across the day (P < 0.0001). Paradoxically, the immediate and delayed satiety quotient of the breakfast also increased significantly (P < 0.05). CONCLUSIONS: These data show that the effect of exercise on appetite regulation involves at least 2 processes: an increase in the overall (orexigenic) drive to eat and a concomitant increase in the satiating efficiency of a fixed meal. PMID- 19675106 TI - Serum vitamin C and the prevalence of vitamin C deficiency in the United States: 2003-2004 National Health and Nutrition Examination Survey (NHANES). AB - BACKGROUND: Vitamin C (ascorbic acid) may be the most important water-soluble antioxidant in human plasma. In the third National Health and Nutrition Examination Survey (NHANES III, 1988-1994), approximately 13% of the US population was vitamin C deficient (serum concentrations <11.4 micromol/L). OBJECTIVE: The aim was to determine the most current distribution of serum vitamin C concentrations in the United States and the prevalence of deficiency in selected subgroups. DESIGN: Serum concentrations of total vitamin C were measured in 7277 noninstitutionalized civilians aged > or =6 y during the cross-sectional, nationally representative NHANES 2003-2004. The prevalence of deficiency was compared with results from NHANES III. RESULTS: The overall age-adjusted mean from the square-root transformed (SM) concentration was 51.4 micromol/L (95% CI: 48.4, 54.6). The highest concentrations were found in children and older persons. Within each race-ethnic group, women had higher concentrations than did men (P < 0.05). Mean concentrations of adult smokers were one-third lower than those of nonsmokers (SM: 35.2 compared with 50.7 micromol/L and 38.6 compared with 58.0 micromol/L in men and women, respectively). The overall prevalence (+/-SE) of age adjusted vitamin C deficiency was 7.1 +/- 0.9%. Mean vitamin C concentrations increased (P < 0.05) and the prevalence of vitamin C deficiency decreased (P < 0.01) with increasing socioeconomic status. Recent vitamin C supplement use or adequate dietary intake decreased the risk of vitamin C deficiency (P < 0.05). CONCLUSIONS: In NHANES 2003-2004, vitamin C status improved, and the prevalence of vitamin C deficiency was significantly lower than that during NHANES III, but smokers and low-income persons were among those at increased risk of deficiency. PMID- 19675107 TI - Differential functional magnetic resonance imaging response to food pictures in successful weight-loss maintainers relative to normal-weight and obese controls. AB - BACKGROUND: Prior research indicates that successful weight-loss maintainers (SWLs) work harder than people of normal weight to maintain their weight loss, including greater dietary restriction of fat and higher physical activity levels. However, little work to date has examined how SWLs differ biologically from normal-weight (NW) and obese controls. OBJECTIVE: The objective was to compare the brain responses of SWLs to food pictures with those of NW and obese controls. DESIGN: Blood oxygen level-dependent responses to high- and low-energy food pictures were measured in 18 NW controls, 16 obese controls, and 17 SWLs. RESULTS: Group differences were identified in 4 regions, which indicated significant change in activation in response to the food pictures. SWLs showed greater activation in the left superior frontal region and right middle temporal region than did NW and obese controls-a pattern of results confirmed in exploratory voxel-wise analyses. Obese controls also showed greater activation in a bilateral precentral region. CONCLUSIONS: These results suggest that SWLs show greater activation in frontal regions and primary and secondary visual cortices-a pattern consistent with greater inhibitory control in response to food cues and greater visual attention to the food cues. A greater engagement of inhibitory control regions in response to food cues as well as a greater monitoring of foods may promote control of food intake and successful weight-loss maintenance. PMID- 19675109 TI - Fruit, vegetables, and colorectal cancer risk: the European Prospective Investigation into Cancer and Nutrition. PMID- 19675108 TI - Dietary correlates of urinary 6-sulfatoxymelatonin concentrations in the Nurses' Health Study cohorts. AB - BACKGROUND: Age and certain lifestyle factors, including a higher body mass index and exposure to light at night, are related to lower circulating concentrations of melatonin-a hormone with probable cancer-protective properties. Although melatonin is a direct derivative of the essential amino acid tryptophan, little is known about the relation of diet with melatonin concentrations. OBJECTIVE: The objective was to examine cross-sectional associations of various nutrients and dietary factors as well as food groups with creatinine-adjusted first morning urinary melatonin (6-sulfatoxymelatonin; aMT6s) concentrations. DESIGN: Participants were 998 healthy women from 2 independent cohorts: the Nurses' Health Study (NHS; n = 585) and NHS II (n = 413). We computed least-squares mean hormone concentrations across categories of dietary variables, with adjustment for total energy intake, age, and other nondietary factors known to be associated with aMT6s concentrations. RESULTS: In multivariate analyses, we found no significant associations between the intake of various nutrients, including tryptophan and urinary melatonin concentrations. A higher intake of meat, particularly red meat, was associated with lower concentrations of aMT6s (adjusted mean concentrations of aMT6s across increasing quartiles of red meat intake were 17.9, 17.0, 18.1, and 15.3 ng/mg creatinine; P for trend = 0.02). In contrast, neither poultry intake (including turkey) nor fish intake was associated with aMT6s concentrations. CONCLUSION: Although no specific nutrients were associated with altered concentrations of melatonin, our findings raise the possibility that several specific foods, including red meat, could affect cancer risk through the lowering of melatonin concentrations. PMID- 19675110 TI - New approach for m-cell-specific molecules screening by comprehensive transcriptome analysis. AB - A minor population of M cells within the follicle-associated epithelium (FAE) of intestinal Peyer's patches (PPs) serves as a major portal for entry of exogenous antigens. Characterization of the mammalian M cells, including identification of M-cell surface molecules used for bacterial uptake, has been hampered by their relative rarity. In contrast, M cells constitute virtually all of the FAE cells in the avian bursa of Fabricius. We therefore performed comparative gene expression profiling of chicken and murine FAE to identify commonly expressed genes by M cells in both species. The comprehensive transcriptome analysis revealed that 28 genes were commonly up-regulated in FAE from both species. In situ hybridization revealed that annexin A10 (Anxa10) mRNA was scattered in FAE, and co-localized with Ulex europaeus agglutinin-1 binding to M cells. Whole-mount immunostaining also revealed that cellular prion protein (PrP(C)) was expressed on the luminal side of the apical plasma membrane of M cells, and co-localized with grycoprotein 2 that recognizes only M cells in murine PP. Our findings identify new M-cell-specific molecules through using comprehensive transcriptome analysis. These conserved molecules in M cells of mice and chickens may play essential roles in M-cell function and/or differentiation. PMID- 19675111 TI - A genome-wide compilation of the two-component systems in Lotus japonicus. AB - The two-component systems (TCS), or histidine-to-aspartate phosphorelays, are evolutionarily conserved common signal transduction mechanisms that are implicated in a wide variety of cellular responses to environmental stimuli in both prokaryotes and eukaryotes including plants. Among higher plants, legumes including Lotus japonicus have a unique ability to engage in beneficial symbiosis with nitrogen-fixing bacteria. We previously presented a genome-wide compiled list of TCS-associated components of Mesorhizobium loti, which is a symbiont specific to L. japonicus (Hagiwara et al. 2004, DNA Res., 11, 57-65). To gain both general and specific insights into TCS of this currently attractive model legume, here we compiled TCS-associated components as many as possible from a genome-wide viewpoint by taking advantage that the efforts of whole genome sequencing of L. japonicus are almost at final stage. In the current database (http://www.kazusa.or.jp/lotus/index.html), it was found that L. japonicus has, at least, 14 genes each encoding a histidine kinase, 7 histidine-containing phosphotransmitter-related genes, 7 type-A response regulator (RR)-related genes, 11 type-B RR-related genes, and also 5 circadian clock-associated pseudo-RR genes. These results suggested that most of the L. japonicus TCS-associated genes have already been uncovered in this genome-wide analysis, if not all. Here, characteristics of these TCS-associated components of L. japonicus were inspected, one by one, in comparison with those of Arabidopsis thaliana. In addition, some critical experiments were also done to gain further insights into the functions of L. japonicus TCS-associated genes with special reference to cytokinin-mediated signal transduction and circadian clock. PMID- 19675112 TI - Do lifestyle or social factors explain ethnic/racial inequalities in breast cancer survival? AB - Despite numerous studies documenting ethnic inequalities in breast cancer survival between minority and majority ethnic groups worldwide, reasons for these inequalities remain unclear. The authors performed a systematic review of published literature to identify studies that investigated the explanatory power of smoking, alcohol consumption, body mass index (BMI), and socioeconomic position (SEP) on ethnic inequalities in breast cancer survival. Sixteen studies were included in the review. From 5 studies, the authors found that differences in breast cancer survival between ethnic groups may be in part explained by BMI, but there was little evidence to implicate smoking or alcohol consumption as explanatory factors of this inequality. From 12 studies, the authors found that SEP explains part of the ethnic inequality in all-cause survival but that it was not evident for breast-cancer-specific survival. SEP explains more of the disparities among African-American versus white women in the United States compared with other ethnic comparisons. Furthermore, given social patterning of BMI and other lifestyle habits, it is possible that results for SEP and BMI are measuring the same effect. In this review, the authors make suggestions regarding the role of epidemiology in facilitating further research to better inform the development of effective policies to address ethnic differences in survival. PMID- 19675113 TI - In memoriam - John Jaques Bullen (1920-2009). PMID- 19675114 TI - How effective are physical activity interventions for alleviating depressive symptoms in older people? A systematic review. AB - BACKGROUND: The benefits of physical exercise in reducing clinically defined depression in the general population have been established, although a review of the evidence for older adults is needed. OBJECTIVES: To assess the efficacy of physical exercise for the treatment of depressive symptoms in older adults (>60 years). DATA SOURCES: We searched: MEDLINE (1966-May 2008); EMBASE (1980-May 2008); Cumulative Index to Nursing & Allied Health Literature (CINAHL; 1982-May 2008); PsycINFO (1966-May 2008), The Cochrane Library (Issue 2, 2008), and National Research Register (NRR; Issue 2, 2008). REVIEW METHODS: Randomized controlled trials and quasi-experimental studies of physical exercise interventions for depression were included where 80% or more of participants were >60 years. Abstracts were assessed to determine whether they met specified inclusion criteria. Primary analysis focused on the prevalence of diagnosable depressive disorder following intervention. Secondary outcome was depression or mood scores on standardized scales. RESULTS: Eleven randomized controlled trials with a total of 641 participants were included in the review. Short-term positive outcome for depression or depressive symptoms was found in nine studies, although the mode, intensity and duration of intervention varied across studies. Medium- to long-term effects of intervention were less clear. CONCLUSION: Physical exercise programmes obtain clinically relevant outcomes in the treatment of depressive symptoms in depressed older people. Exercise, though not appropriate for all in this population, may improve mood in this group. Further research is needed to establish medium- to long-term effects and cost-effectiveness. PMID- 19675115 TI - Effectiveness of a video-based exercise programme to reduce falls and improve health-related quality of life among older adults discharged from hospital: a pilot randomized controlled trial. AB - OBJECTIVE: Falls, loss of health-related quality of life and physical capacity, reduced participation in activities of daily living, and increased fear of falling are all potential outcomes for older adults discharged from hospital. A low-cost video based exercise programme may address this. DESIGN: This study was a randomized controlled trial with blinded outcomes assessment and a six-month follow-up. SUBJECTS AND SETTING: Participants were older adults (>65 years) using a mobility aid discharged from a tertiary hospital in Brisbane, Australia, without referral for community-based rehabilitation services. INTERVENTION: A digital video disk-based programme encompassing six exercise types each with six levels of difficulty. A home visit from a project physiotherapist was conducted to ensure patient safety. Control group patients received usual care. MAIN MEASURES: Falls, health-related quality of life, participation in activities of daily living, physical capacity and fear of falling. RESULTS: Study participants (n = 53, 19 intervention, 34 control) experienced decreasing health-related quality of life, several falls (72), and lower levels of participation in activities of daily living over the six-month follow-up. The intervention group did not differ significantly from the control group in terms of the outcomes examined, though a non-significant reduction in the rate of falls was observed. Intervention group participants complied with the exercise programme well during the first two weeks following discharge from hospital but then reduced their compliance levels thereafter. CONCLUSIONS: The intervention may be beneficial for reducing the rate of falls in this patient population though further research with a larger sample size is indicated. PMID- 19675117 TI - Coping with Meniere's disease: experience and benefits from the use of complementary and alternative medicine. AB - OBJECTIVES: To identify the significant events and persons prompting search for non-biomedical/complementary and alternative (CAM) ways to cope with the impact of Meniere's syndrome, the CAM therapies used and perceived benefits. METHODS: A narrative-based approach, with participants recruited from web sites and newsletters of two Meniere's Societies (in the UK and in Victoria, Australia) and searches of a monthly newsletter, Spin, for letters related to the use of CAM therapies. A thematic approach to data analysis was undertaken. RESULTS: Twenty personal, written stories were obtained and 23 letters. Commonly, critical events or changes in the condition provided the catalyst to seek alternative treatment, and sometimes significant others (a GP, CAM practitioner, friends, family). The accounts illustrate a range and variety of journeys that participants travelled in locating satisfactory ways to manage the condition, and the essential 'personal' nature of the experience and perceived benefits. No single CAM solution or pathway was evident. DISCUSSION: The findings suggest the importance of 'finding your own way', having a positive outlook and being proactive in regaining control over one's health, and the value of early recommendations to 'perhaps try alternative/ CAM treatments'. PMID- 19675118 TI - Gender differences in sense of coherence, perceived social support, and negative emotions among drug-abstinent israeli inmates. AB - This study examines gender differences in the sense of coherence, perceived social support, and negative emotions among drug-abstinent Israeli inmates. One hundred nineteen inmates have participated in this study (65 men and 54 women). The findings indicate that among female inmates, abstinence of more than a year is related to a decrease in sense of coherence and to an increase in their perceived friend support. In contrast, among male inmates, abstinence of more than a year is related to an increase in sense of coherence and no change at all in their perceived friend support. In addition, hostility level is lower and sense of coherence is higher when the length of abstinence exceeds a year among male inmates. In contrast, among female inmates hostility level is higher when the sense of coherence is lower, without any connection to the length of abstinence. The implications of these findings to treatment intervention are discussed. PMID- 19675116 TI - Physicians' participatory decision-making and quality of diabetes care processes and outcomes: results from the triad study. AB - OBJECTIVES: In participatory decision-making (PDM), physicians actively engage patients in treatment and other care decisions. Patients who report that their physicians engage in PDM have better disease self-management and health outcomes. We examined whether physicians' diabetes-specific treatment PDM preferences as well as their self-reported practices are associated with the quality of diabetes care their patients receive. METHODS: 2003 cross-sectional survey and medical record review of a random sample of diabetes patients (n=4198) in 10 US health plans across the country and their physicians (n=1217). We characterized physicians' diabetes care PDM preferences and practices as 'no patient involvement,' 'physician-dominant,' 'shared,' or 'patient-dominant' and conducted multivariate analyses examining their effects on the following: (1) three diabetes care processes (annual hemoglobin A1c test; lipid test; and dilated retinal exam); (2) patients'satisfaction with physician communication; and (3) whether patients' A1c, systolic blood pressure (SBP), and low-density lipoprotein cholesterol (LDL) were in control. RESULTS: Most physicians preferred 'shared' PDM (58%) rather than 'no patient involvement' (9%), 'physician-dominant' (28%) or 'patient dominant' PDM (5%). However, most reported practicing 'physician dominant' PDM (43%) with most of their patients, rather than 'no patient involvement' (13%), 'shared' (37%) or 'patient-dominant' PDM (7%). After adjusting for patient and physician-level characteristics and clustering by health plan, patients of physicians who preferred 'shared' PDM were more likely to receive A1c tests [90% vs. 82%, AOR: 2.05, 95% CI: 1.03-3.07] and patients of physicians who preferred 'patient-dominant' treatment decision-making were more likely to receive lipid tests [60% vs. 50%, AOR: 1.58, 95% CI: 1.04-2.39] than those of providers who preferred 'no patient involvement' in treatment decision making. There were no differences in patients' satisfaction with their doctor's communication or control of A1c, SBP or LDL depending on their physicians' PDM preferences. Physicians' self-reported PDM practices were not associated with any of the examined aspects of diabetes care in multivariate analyses. CONCLUSIONS: Patients whose physicians prefer more patient involvement in decision-making are more likely than patients whose physicians prefer more physician-directed styles to receive some recommended risk factor screening tests, an important first step toward improved diabetes outcomes. Involving patients in treatment decision making alone, however, appears not to be sufficient to improve biomedical outcomes. PMID- 19675119 TI - Differential modulation of human {beta}-defensins expression in human gingival epithelia by Porphyromonas gingivalis lipopolysaccharide with tetra- and penta acylated lipid A structures. AB - Porphyromonas gingivalis lipopolysaccharide (LPS) is a crucial virulence factor strongly involved in the development of chronic periodontitis. It displays a significant amount of lipid A structural heterogeneity, containing both tetra- (LPS(1435/1449) ) and penta-acylated (LPS( 1690)) lipid A structures with opposing effects on E-selectin expression in human endothelial cells. Little is known about how these two isoforms of P. gingivalis LPS could differentially affect host innate immune responses in human gingival epithelia. The present study compares the modulatory effects of P. gingivalis LPS(1435/1449) and LPS(1690) on the expression of human beta-defensins (hBDs) in the reconstituted human gingival epithelium, and examines the involvements of a panel of pattern recognition receptors in the modulatory effects concerned. It is shown that hBD 1, hBD-2 and hBD-3 mRNAs are significantly up-regulated by P. gingivalis LPS(1690), but down-regulated by P. gingivalis LPS( 1435/1449). Toll-like receptor (TLR) 2 and CD14 mRNAs are also differentially regulated, and the modulation of hBD-2 expression may be through the co-operation of both TLR2 and TLR4. This study suggests that P. gingivalis LPS with different lipid A structures could differentially modulate host innate immune responses in human gingival epithelia, which may be a hitherto undescribed novel pathogenic mechanism of P. gingivalis in periodontal pathogenesis. PMID- 19675120 TI - Altered innate immune response in farmers and smokers. AB - Pig farmers and cigarette smokers are continuously exposed to pathogen-associated molecular patterns (PAMPs) have an increased prevalence of respiratory disorders, such as chronic bronchitis and chronic obstructive pulmonary decease (COPD). We hypothesized that markers of innate immunity, T-helper (Th) cell cytokine profile and acute responses to pro-inflammatory stimuli differ between smokers and farmers, who are exposed to organic material on a daily basis and healthy non exposed subjects. Eleven non-smoking pig farmers, 12 non-farming smokers and 12 controls underwent bronchial lipopolysaccharide (LPS) challenge and exposure in a pig barn during 3 h on separate days. Toll-like receptor 2 (TLR2), TLR4 and CD14 on blood monocytes and neutrophils and intracellular cytokine profile of Th cells were assessed before and 7 h after exposures. The same outcomes were analysed on peripheral blood and purified neutrophils from farmers and controls after stimulation ex vivo with dust from a pig barn and LPS. Circulating neutrophils and IL-13 and IL-4 producing Th cells were increased in smokers and farmers and TLR2 expression on blood monocytes was decreased in farmers compared with controls and smokers. After in vivo exposure, altered TLR expression was only observed in controls and the ex vivo stimulations showed an attenuated response in farmers compared to the control group. The inflammatory systemic response to pro-inflammatory stimuli is altered in farmers and smokers probably because of adaptive mechanisms arising from chronic exposure to organic material. This increased proportion of Th2 cells and reduced TLR2 expression may have health related implications and may be related to the increased prevalence of respiratory disorders observed in these groups. PMID- 19675121 TI - The effects of a secondary task on forward and backward walking in Parkinson's disease. AB - BACKGROUND: People with Parkinson's disease (PD) often fall while multitasking or walking backward, unavoidable activities in daily living. Dual tasks involving cognitive demand during gait and unfamiliar motor skills, such as backward walking, could identify those with fall risk, but dual tasking while walking backward has not been examined in those with PD, those who experience freezing of gait (FOG), or healthy older controls. METHODS: A total of 78 people with PD (mean age = 65.1 +/-9.5 years; female, 28%) and 74 age-matched and sex-matched controls (mean age = 65.0 +/-10.0 years; female, 23%) participated. A computerized walkway measured gait velocity, stride length, swing percent, stance percent, cadence, heel to heel base of support, functional ambulation profile, and gait asymmetry during forward and backward walking with and without a secondary cognitive task. RESULTS: Direction and task effects on walking performance were similar between healthy controls and those with PD. However, those with PD were more affected than controls, and freezers were more affected than nonfreezers, by backward walking and dual tasking. Walking backward seemed to affect gait more than dual tasking in those with PD,although the subset of freezers appeared particularly affected by both challenges. CONCLUSION: People with PD are impaired while performing complex motor and mental tasks simultaneously,which may put them at risk for falling. Those with FOG are more adversely affected by both motor and mental challenges than those without. Evaluation of backward walking while performing a secondary task might be an effective clinical tool to identify locomotor difficulties. PMID- 19675122 TI - The effects of repeated rehabilitation "tune-ups" on functional recovery after focal ischemia in rats. AB - BACKGROUND: For most stroke survivors, rehabilitation therapy is the only treatment option available. The beneficial effects of early rehabilitation on neuroplasticity and functional recovery have been modeled in experimental stroke using a combination of enriched environment and rehabilitation. However, the impact of a secondary intervention, such as a periodic return to therapy, remains unclear. OBJECTIVE: This study examines whether a return to enriched rehabilitation (ie, "tune-up") can further promote functional recovery or produce beneficial changes in brain plasticity in the chronic phase of stroke recovery. METHODS: Rats were exposed to focal ischemia (endothelin-1 applied to forelimb sensorimotor cortex and dorsolateral striatum) and allowed to recover either in standard housing or in a combination of enriched environment and rehabilitative reaching for 9 weeks. Animals were then exposed to rotating periods of standard housing (5 weeks) and intensive "tune-up" therapy consisting of various sensorimotor/cognitive activities (2 weeks). Functional recovery was assessed using the Montoya staircase, beam-traversing, and cylinder tests, and Golgi-Cox analysis was used to examine dendritic complexity in the contralesional forelimb motor cortex. RESULTS: Although early enriched rehabilitation significantly improved sensorimotor function in both the beam and staircase tests, "tune-up" therapy had no effect on recovery. Golgi-Cox analysis revealed no effect of treatment on dendritic complexity. CONCLUSIONS: This study reaffirms the benefits of early rehabilitation for functional recovery after stroke. However, "tune-up" therapy provided no benefit in ischemic animals regardless of earlier rehabilitation experience. It is possible that alternative approaches in the chronic phase may prove more effective. PMID- 19675123 TI - Falls in persons with spinal cord injury: validity and reliability of the Berg Balance Scale. AB - BACKGROUND: Persons with spinal cord injury who are able to walk are at risk for falls. OBJECTIVE: The objectives were to investigate if the Berg Balance Scale (BBS) can discriminate those with a propensity to fall; to determine whether the BBS is associated with mobility measures, fear of falling, and muscle strength; and to assess interobserver reliability. METHODS: The measurement tools used were the BBS, the Spinal Cord Independence Measure, the Falls Efficacy Scale (FES-I), the Walking Index for Spinal Cord Injury, the 10-m walk test, and the standard neurological classification including motor scores (MS). Falls were recorded retrospectively for the previous month and prospectively for the subsequent 4 months. To determine interobserver reliability, BBS performance was videotaped and analyzed by additional physical therapists. Associations between BBS and the number of falls, measures of mobility, FES-I, and MS were calculated using Spearman correlations. The interobserver reliability was quantified using Kendall's coefficient of concordance and intraclass correlation coefficients (ICCs). RESULTS: Forty-two participants were included of whom 26 sustained 1 or more falls. BBS performance correlated with measures of mobility, FES-I, and MS (r(s) = -.83 to .93; P < .001) but not with the number of falls (r(s) = -.17; P = .28). The interobserver reliability was excellent, both for single items (.84 .98, P < .001) and for the total score (ICC = .95; 95% confidence interval = 0.910-0.975). CONCLUSIONS: The BBS proved to be reliable and to relate well with other mobility measures, fear of falling, and muscle strength. However, it was unable discriminate between people who did fall and people who did not fall. PMID- 19675124 TI - Comment on "Evidence of increased motoneuron excitability in stroke patients without clinical spasticity". PMID- 19675125 TI - Direct detection of salmonella cells in the air of livestock stables by real-time PCR. AB - A SYBR Green real-time quantitative polymerase chain reaction (qPCR) assay for specific detection and quantification of airborne Salmonella cells in livestock housings is presented. A set of specific primers was tested and validated for specific detection and quantification of Salmonella-specific invA genes of DNA extracted from bioaerosol samples. Application of the method to poultry house bioaerosol samples showed concentrations ranging from 2.2 x 10(1) to 3 x 10(6) Salmonella targets m(-3) of air. Salmonella were also detected by a cultivation based approach in some samples, but concentrations were two to three magnitudes lower than the concentrations detected by molecular biological results. Specificity of results was demonstrated by cloning analyses of PCR products, which were exclusively assigned to the genus Salmonella. However, by molecular methods, microorganisms are detected independently of their viability status, leading to an overestimation of concentration. Hence, the survival rate of Salmonella cells was measured on filter surfaces during filtration samplings where 82% of the cells died within 20 min of filtration. The results clearly show the specificity and practicability of the established qPCR assay for analysis and quantification of salmonellae in bioaerosols. The results demonstrate airborne Salmonella workplace concentrations in poultry production of up to 3.3% of 4',6 Diamidino-2-phenylindole-counted total cell numbers. PMID- 19675126 TI - Myosin II regulation during C. elegans embryonic elongation: LET-502/ROCK, MRCK-1 and PAK-1, three kinases with different roles. AB - Myosin II plays a central role in epithelial morphogenesis; however, its role has mainly been examined in processes involving a single cell type. Here we analyze the structure, spatial requirement and regulation of myosin II during C. elegans embryonic elongation, a process that involves distinct epidermal cells and muscles. We developed novel GFP probes to visualize the dynamics of actomyosin remodeling, and found that the assembly of myosin II filaments, but not actin microfilaments, depends on the myosin regulatory light chain (MLC-4) and essential light chain (MLC-5, which we identified herein). To determine how myosin II regulates embryonic elongation, we rescued mlc-4 mutants with various constructs and found that MLC-4 is essential in a subset of epidermal cells. We show that phosphorylation of two evolutionary conserved MLC-4 serine and threonine residues is important for myosin II activity and organization. Finally, in an RNAi screen for potential myosin regulatory light chain kinases, we found that the ROCK, PAK and MRCK homologs act redundantly. The combined loss of ROCK and PAK, or ROCK and MRCK, completely prevented embryonic elongation, but a constitutively active form of MLC-4 could only rescue a lack of MRCK. This result, together with systematic genetic epistasis tests with a myosin phosphatase mutation, suggests that ROCK and MRCK regulate MLC-4 and the myosin phosphatase. Moreover, we suggest that ROCK and PAK regulate at least one other target essential for elongation, in addition to MLC-4. PMID- 19675127 TI - Systematic analysis of dynamic miRNA-target interactions during C. elegans development. AB - Although microRNA (miRNA)-mediated functions have been implicated in many aspects of animal development, the majority of miRNA::mRNA regulatory interactions remain to be characterized experimentally. We used an AIN/GW182 protein immunoprecipitation approach to systematically analyze miRNA::mRNA interactions during C. elegans development. We characterized the composition of miRNAs in functional miRNA-induced silencing complexes (miRISCs) at each developmental stage and identified three sets of miRNAs with distinct stage-specificity of function. We then identified thousands of miRNA targets in each developmental stage, including a significant portion that is subject to differential miRNA regulation during development. By identifying thousands of miRNA family-mRNA pairs with temporally correlated patterns of AIN-2 association, we gained valuable information on the principles of physiological miRNA::target recognition and predicted 1589 high-confidence miRNA family::mRNA interactions. Our data support the idea that miRNAs preferentially target genes involved in signaling processes and avoid genes with housekeeping functions, and that miRNAs orchestrate temporal developmental programs by coordinately targeting or avoiding genes involved in particular biological functions. PMID- 19675128 TI - Vegetally localized Xenopus trim36 regulates cortical rotation and dorsal axis formation. AB - Specification of the dorsoventral axis in Xenopus depends on rearrangements of the egg vegetal cortex following fertilization, concomitant with activation of Wnt/beta-catenin signaling. How these processes are tied together is not clear, but RNAs localized to the vegetal cortex during oogenesis are known to be essential. Despite their importance, few vegetally localized RNAs have been examined in detail. In this study, we describe the identification of a novel localized mRNA, trim36, and characterize its function through maternal loss-of function experiments. We find that trim36 is expressed in the germ plasm and encodes a ubiquitin ligase of the Tripartite motif-containing (Trim) family. Depletion of maternal trim36 using antisense oligonucleotides results in ventralized embryos and reduced organizer gene expression. We show that injection of wnt11 mRNA rescues this effect, suggesting that Trim36 functions upstream of Wnt/beta-catenin activation. We further find that vegetal microtubule polymerization and cortical rotation are disrupted in trim36-depleted embryos, in a manner dependent on Trim36 ubiquitin ligase activity. Additionally, these embryos can be rescued by tipping the eggs 90 degrees relative to the animal vegetal axis. Taken together, our results suggest a role for Trim36 in controlling the stability of proteins regulating microtubule polymerization during cortical rotation, and subsequently axis formation. PMID- 19675130 TI - Enhancer-promoter communication at the Drosophila engrailed locus. AB - Enhancers are often located many tens of kilobases away from the promoter they regulate, sometimes residing closer to the promoter of a neighboring gene. How do they know which gene to activate? We have used homing P[en] constructs to study the enhancer-promoter communication at the engrailed locus. Here we show that engrailed enhancers can act over large distances, even skipping over other transcription units, choosing the engrailed promoter over those of neighboring genes. This specificity is achieved in at least three ways. First, early acting engrailed stripe enhancers exhibit promoter specificity. Second, a proximal promoter-tethering element is required for the action of the imaginal disc enhancer(s). Our data suggest that there are two partially redundant promoter tethering elements. Third, the long-distance action of engrailed enhancers requires a combination of the engrailed promoter and sequences within or closely linked to the promoter proximal Polycomb-group response elements. These data show that multiple mechanisms ensure proper enhancer-promoter communication at the Drosophila engrailed locus. PMID- 19675129 TI - A chromatin insulator mediates transgene homing and very long-range enhancer promoter communication. AB - Insulator sequences help to organize the genome into discrete functional regions by preventing inappropriate cross-regulation. This is thought to be mediated in part through associations with other insulators located elsewhere in the genome. Enhancers that normally drive Drosophila even skipped (eve) expression are located closer to the TER94 transcription start site than to that of eve. We discovered that the region between these genes has enhancer-blocking activity, and that this insulator region also mediates homing of P-element transgenes to the eve-TER94 genomic neighborhood. Localization of these activities to within 0.6 kb failed to separate them. Importantly, homed transgenic promoters respond to endogenous eve enhancers from great distances, and this long-range communication depends on the homing/insulator region, which we call Homie. We also find that the eve promoter contributes to long-distance communication. However, even the basal hsp70 promoter can communicate with eve enhancers across distances of several megabases, when the communication is mediated by Homie. These studies show that, while Homie blocks enhancer-promoter communication at short range, it facilitates long-range communication between distant genomic regions, possibly by organizing a large chromosomal loop between endogenous and transgenic Homies. PMID- 19675131 TI - Receptor tyrosine phosphatases control tracheal tube geometries through negative regulation of Egfr signaling. AB - The formation of epithelial tubes with defined shapes and sizes is essential for organ development. We describe a unique tracheal tubulogenesis phenotype caused by loss of both Drosophila type III receptor tyrosine phosphatases (RPTPs), Ptp4E and Ptp10D. Ptp4E is the only widely expressed Drosophila RPTP, and is the last of the six fly RPTPs to be genetically characterized. We recently isolated mutations in Ptp4E, and discovered that, although Ptp4E null mutants have no detectable phenotypes, double mutants lacking both Ptp4E and Ptp10D display synthetic lethality at hatching owing to respiratory failure. In these double mutants, unicellular and terminal tracheal branches develop large bubble-like cysts that selectively incorporate apical cell surface markers. Cysts in unicellular branches are enlargements of the lumen that are sealed by adherens junctions, whereas cysts in terminal branches are cytoplasmic vacuoles. Cyst size and number are increased by tracheal expression of activated Egfr tyrosine kinase, and decreased by reducing Egfr levels. Ptp10D forms a complex with Egfr in transfected cells. Downregulation of Egfr signaling by the RPTPs is required for the construction of tubular lumens, whether extracellular or intracellular, by cells that undergo remodeling during branch morphogenesis. The Ptp4E Ptp10D phenotype represents the first evidence of an essential role for RPTPs in epithelial organ development. These findings might be relevant to organ development and disease in mammals, because PTPRJ (DEP-1), an ortholog of Ptp4E/Ptp10D, interacts with the hepatocyte growth factor receptor tyrosine kinase. PTPRJ corresponds to the murine Scc1 (suppressor of colon cancer) gene. PMID- 19675132 TI - Abi plays an opposing role to Abl in Drosophila axonogenesis and synaptogenesis. AB - Abl tyrosine kinase (Abl) regulates axon guidance by modulating actin dynamics. Abelson interacting protein (Abi), originally identified as a kinase substrate of Abl, also plays a key role in actin dynamics, yet its role with respect to Abl in the developing nervous system remains unclear. Here we show that mutations in abi disrupt axonal patterning in the developing Drosophila central nervous system (CNS). However, reducing abi gene dosage by half substantially rescues Abl mutant phenotypes in pupal lethality, axonal guidance defects and locomotion deficits. Moreover, we show that mutations in Abl increase synaptic growth and spontaneous synaptic transmission frequency at the neuromuscular junction. Double heterozygosity for abi and enabled (ena) also suppresses the synaptic overgrowth phenotypes of Abl mutants, suggesting that Abi acts cooperatively with Ena to antagonize Abl function in synaptogenesis. Intriguingly, overexpressing Abi or Ena alone in cultured cells dramatically redistributed peripheral F-actin to the cytoplasm, with aggregates colocalizing with Abi and/or Ena, and resulted in a reduction in neurite extension. However, co-expressing Abl with Abi or Ena redistributed cytoplasmic F-actin back to the cell periphery and restored bipolar cell morphology. These data suggest that abi and Abl have an antagonistic interaction in Drosophila axonogenesis and synaptogenesis, which possibly occurs through the modulation of F-actin reorganization. PMID- 19675133 TI - Vascular endothelial growth factor inhibition by dRK6 causes endothelial apoptosis, fibrosis, and inflammation in the heart via the Akt/eNOS axis in db/db mice. AB - OBJECTIVE: Vascular endothelial growth factor (VEGF), which is associated with the stimulation of angiogenesis and collateral vessel synthase, is one of the crucial factors involved in cardiac remodeling in type 2 diabetes. RESEARCH DESIGN AND METHODS: We investigated VEGF inhibition by dRK6 on the heart in an animal model of type 2 diabetes. Male db/db and db/m mice either were treated with dRK6 starting at 7 weeks of age for 12 weeks (db/db-dRK6 and db/m-dRK6) or were untreated. RESULTS: Cardiac dysfunction and hypertrophy were noted by echocardiogram and molecular markers in the db/db-dRK6 mice. The presence of diabetes significantly suppressed the expression of VEGF receptor (VEGFR)-1 and VEGFR-2, phospho-Akt, and phospho-endothelial nitric oxide synthase (eNOS) in the heart. In db/db-dRK6 mice, dRK6 completely inhibited VEGFR-2, phospho-Akt, and phospho-eNOS expression, whereas no effect on VEGFR-1 was observed. Cardiac fibrosis, microvascular scarcity associated with an increase in apoptotic endothelial cells, and inflammation were prominent, as well as increase in antiangiogenic growth factors. Cardiac 8-hydroxy-deoxyguanine and hypoxia inducible factor-1alpha expression were significantly increased. No such changes were found in the other groups, including the db/m-dRK6 mice. The number of apoptotic human umbilical vein endothelial cells was increased by dRK6 in a dose dependent manner only at high glucose concentrations, and this was associated with a decrease in phospho-Akt and phospho-eNOS related to oxidative stress. CONCLUSIONS: Our results demonstrated that systemic blockade of VEGF by dRK6 had deleterious effects on the heart in an animal model of type 2 diabetes; dRK6 induced downregulation of the VEGFR-2 and Akt-eNOS axis and enhancement of oxidative stress. PMID- 19675134 TI - Muscle contraction, but not insulin, increases microvascular blood volume in the presence of free fatty acid-induced insulin resistance. AB - OBJECTIVE: Insulin and contraction each increase muscle microvascular blood volume (MBV) and glucose uptake. Inhibiting nitric oxide synthase blocks insulin's but not contraction's effects. We examined whether contraction could augment the MBV increase seen with physiologic hyperinsulinemia and whether free fatty acid (FFA)-induced insulin resistance differentially affects contraction- versus insulin-mediated increases in MBV. RESEARCH DESIGN AND METHODS: Rats were fasted overnight. Plasma FFAs were increased by intralipid/heparin infusion (3 h), insulin was increased with a euglycemic clamp (3 mU x min(-1) x kg(-1)), and hindlimb muscle contraction was electrically stimulated. Muscle MBV was measured using contrast-enhanced ultrasound. Insulin transport into muscle was measured using (125)I-insulin. BQ-123 (0.4 mg/h) was used to block the endothelin-1 (ET-1) receptor A. RESULTS: Superimposing contraction on physiologic hyperinsulinemia increased MBV within 10 min by 37 and 67% for 0.1 or 1 Hz, respectively (P < 0.01). FFA elevation alone did not affect MBV, whereas 0.1 Hz stimulation doubled MBV (P < 0.05) and increased muscle insulin uptake (P < 0.05) despite high FFA. Physiologic hyperinsulinemia during FFA elevation paradoxically decreased MBV (P < 0.05). This MBV decrease was reversed by either 0.1 Hz contraction or ET-1 receptor A antagonism, and the combination raised MBV above basal. CONCLUSIONS: Contraction recruits microvasculature beyond that seen with physiologic hyperinsulinemia by a distinct mechanism that is not blocked by FFA-induced vascular insulin resistance. The paradoxical MBV decline seen with insulin plus FFA may result from differential inhibition of insulin-stimulated nitric oxide dependent vasodilation relative to ET-1 vasoconstriction. Our results implicate ET-1 as a potential mediator of FFA-induced vascular insulin resistance. PMID- 19675135 TI - Validity and reproducibility of measurement of islet autoreactivity by T-cell assays in subjects with early type 1 diabetes. AB - OBJECTIVE: Type 1 diabetes results from an immunemediated destruction of beta cells, likely to be mediated by T lymphocytes, but the sensitivity, specificity, and other measures of validity of existing assays for islet autoreactive T-cells are not well established. Such assays are vital for monitoring responses to interventions that may modulate disease progression. RESEARCH DESIGN AND METHODS: We studied the ability of cellular assays to discriminate responses in patients with type 1 diabetes and normal control subjects in a randomized blinded study in the U.S. and U.K. We evaluated the reproducibility of these measurements overall and to individual analytes from repeat collections. RESULTS: Responses in the cellular immunoblot, U.K.-ELISPOT, and T-cell proliferation assays could differentiate patients from control subjects with odds ratios of 21.7, 3.44, and 3.36, respectively, with sensitivity and specificity as high as 74 and 88%. The class II tetramer and U.S. ELISPOT assays performed less well. Despite the significant association of the responses with type 1 diabetes, the reproducibility of the measured responses, both overall and individual analytes, was relatively low. Positive samples from normal control subjects (i.e., false positives) were generally isolated to single assays. CONCLUSIONS: The cellular immunoblot, U.K.-ELISPOT, and T-cell proliferation assays can distinguish responses from patients with type 1 diabetes and healthy control subjects. The limited reproducibility of the measurements overall and of responses to individual analytes may reflect the difficulty in detection of low frequency of antigen-specific T-cells or variability in their appearance in peripheral blood. PMID- 19675136 TI - Paradoxical coupling of triglyceride synthesis and fatty acid oxidation in skeletal muscle overexpressing DGAT1. AB - OBJECTIVE: Transgenic expression of diacylglycerol acyltransferase-1 (DGAT1) in skeletal muscle leads to protection against fat-induced insulin resistance despite accumulation of intramuscular triglyceride, a phenomenon similar to what is known as the "athlete paradox." The primary objective of this study is to determine how DGAT1 affects muscle fatty acid oxidation in relation to whole-body energy metabolism and insulin sensitivity. RESEARCH DESIGN AND METHODS: We first quantified insulin sensitivity and the relative tissue contributions to the improved whole-body insulin sensitivity in muscle creatine kisase (MCK)-DGAT1 transgenic mice by hyperinsulinemic-euglycemic clamps. Metabolic consequences of DGAT1 overexpression in skeletal muscles were determined by quantifying triglyceride synthesis/storage (anabolic) and fatty acid oxidation (catabolic), in conjunction with gene expression levels of representative marker genes in fatty acid metabolism. Whole-body energy metabolism including food consumption, body weights, oxygen consumption, locomotor activity, and respiration exchange ratios were determined at steady states. RESULTS: MCK-DGAT1 mice were protected against muscle lipoptoxicity, although they remain susceptible to hepatic lipotoxicity. While augmenting triglyceride synthesis, DGAT1 overexpression also led to increased muscle mitochondrial fatty acid oxidation efficiency, as compared with wild-type muscles. On a high-fat diet, MCK-DGAT1 mice displayed higher basal metabolic rates and 5-10% lower body weights compared with wild-type littermates, whereas food consumption was not different. CONCLUSIONS: DGAT1 overexpression in skeletal muscle led to parallel increases in triglyceride synthesis and fatty acid oxidation. Seemingly paradoxical, this phenomenon is characteristic of insulin-sensitive myofibers and suggests that DGAT1 plays an active role in metabolic "remodeling" of skeletal muscle coupled with insulin sensitization. PMID- 19675137 TI - Adipose tissue exosome-like vesicles mediate activation of macrophage-induced insulin resistance. AB - OBJECTIVE: We sought to determine whether exosome-like vesicles (ELVs) released from adipose tissue play a role in activation of macrophages and subsequent development of insulin resistance in a mouse model. RESEARCH DESIGN AND METHODS: ELVs released from adipose tissue were purified by sucrose gradient centrifugation and labeled with green fluorescent dye and then intravenously injected into B6 ob/ob mice (obese model) or B6 mice fed a high-fat diet. The effects of injected ELVs on the activation of macrophages were determined through analysis of activation markers by fluorescence-activated cell sorter and induction of inflammatory cytokines using an ELISA. Glucose tolerance and insulin tolerance were also evaluated. Similarly, B6 mice with different gene knockouts including TLR2, TLR4, MyD88, and Toll-interleukin-1 receptor (TIR) domain containing adaptor protein inducing interferon-beta (TRIF) were also used for testing their responses to the injected ELVs. RESULTS: ELVs are taken up by peripheral blood monocytes, which then differentiate into activated macrophages with increased secretion of tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6). Injection of obELVs into wild-type C57BL/6 mice results in the development of insulin resistance. When the obELVs were intravenously injected into TLR4 knockout B6 mice, the levels of glucose intolerance and insulin resistance were much lower. RBP4 is enriched in the obELVs. Bone marrow derived macrophages preincubated with recombinant RBP4 led to attenuation of obELV-mediated induction of IL-6 and TNF-alpha. CONCLUSIONS: ELVs released by adipose tissue can act as a mode of communication between adipose tissues and macrophages. The obELV-mediated induction of TNF-alpha and IL-6 in macrophages and insulin resistance requires the TLR4/TRIF pathway. PMID- 19675139 TI - C. elegans as model for the study of high glucose- mediated life span reduction. AB - OBJECTIVE: Establishing Caenorhabditis elegans as a model for glucose toxicity mediated life span reduction. RESEARCH DESIGN AND METHODS: C. elegans were maintained to achieve glucose concentrations resembling the hyperglycemic conditions in diabetic patients. The effects of high glucose on life span, glyoxalase-1 activity, advanced glycation end products (AGEs), and reactive oxygen species (ROS) formation and on mitochondrial function were studied. RESULTS: High glucose conditions reduced mean life span from 18.5 + or - 0.4 to 16.5 + or - 0.6 days and maximum life span from 25.9 + or - 0.4 to 23.2 + or - 0.4 days, independent of glucose effects on cuticle or bacterial metabolization of glucose. The formation of methylglyoxal-modified mitochondrial proteins and ROS was significantly increased by high glucose conditions and reduced by mitochondrial uncoupling and complex IIIQo inhibition. Overexpression of the methylglyoxal-detoxifying enzyme glyoxalase-1 attenuated the life-shortening effect of glucose by reducing AGE accumulation (by 65%) and ROS formation (by 50%) and restored mean (16.5 + or - 0.6 to 20.6 + or - 0.4 days) and maximum life span (23.2 + or - 0.4 to 27.7 + or - 2.3 days). In contrast, inhibition of glyoxalase-1 by RNAi further reduced mean (16.5 + or - 0.6 to 13.9 + or - 0.7 days) and maximum life span (23.2 + or - 0.4 to 20.3 + or - 1.1 days). The life span reduction by glyoxalase-1 inhibition was independent from the insulin signaling pathway because high glucose conditions also affected daf-2 knockdown animals in a similar manner. CONCLUSIONS: C. elegans is a suitable model organism to study glucose toxicity, in which high glucose conditions limit the life span by increasing ROS formation and AGE modification of mitochondrial proteins in a daf-2 independent manner. Most importantly, glucose toxicity can be prevented by improving glyoxalase-1-dependent methylglyoxal detoxification or preventing mitochondrial dysfunction. PMID- 19675138 TI - 11beta-hydroxysteroid dehydrogenase type 1 regulates glucocorticoid-induced insulin resistance in skeletal muscle. AB - OBJECTIVE: Glucocorticoid excess is characterized by increased adiposity, skeletal myopathy, and insulin resistance, but the precise molecular mechanisms are unknown. Within skeletal muscle, 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) converts cortisone (11-dehydrocorticosterone in rodents) to active cortisol (corticosterone in rodents). We aimed to determine the mechanisms underpinning glucocorticoid-induced insulin resistance in skeletal muscle and indentify how 11beta-HSD1 inhibitors improve insulin sensitivity. RESEARCH DESIGN AND METHODS: Rodent and human cell cultures, whole-tissue explants, and animal models were used to determine the impact of glucocorticoids and selective 11beta HSD1 inhibition upon insulin signaling and action. RESULTS: Dexamethasone decreased insulin-stimulated glucose uptake, decreased IRS1 mRNA and protein expression, and increased inactivating pSer(307) insulin receptor substrate (IRS) 1. 11beta-HSD1 activity and expression were observed in human and rodent myotubes and muscle explants. Activity was predominantly oxo-reductase, generating active glucocorticoid. A1 (selective 11beta-HSD1 inhibitor) abolished enzyme activity and blocked the increase in pSer(307) IRS1 and reduction in total IRS1 protein after treatment with 11DHC but not corticosterone. In C57Bl6/J mice, the selective 11beta-HSD1 inhibitor, A2, decreased fasting blood glucose levels and improved insulin sensitivity. In KK mice treated with A2, skeletal muscle pSer(307) IRS1 decreased and pThr(308) Akt/PKB increased. In addition, A2 decreased both lipogenic and lipolytic gene expression. CONCLUSIONS: Prereceptor facilitation of glucocorticoid action via 11beta-HSD1 increases pSer(307) IRS1 and may be crucial in mediating insulin resistance in skeletal muscle. Selective 11beta-HSD1 inhibition decreases pSer(307) IRS1, increases pThr(308) Akt/PKB, and decreases lipogenic and lipolytic gene expression that may represent an important mechanism underpinning their insulin-sensitizing action. PMID- 19675140 TI - Caveolin-1 and altered neuregulin signaling contribute to the pathophysiological progression of diabetic peripheral neuropathy. AB - OBJECTIVE: Evaluate if Erb B2 activation and the loss of caveolin-1 (Cav1) contribute to the pathophysiological progression of diabetic peripheral neuropathy (DPN). RESEARCH DESIGN AND METHODS: Cav1 knockout and wild-type C57BL/6 mice were rendered diabetic with streptozotocin, and changes in motor nerve conduction velocity (MNCV), mechanical and thermal hypoalgesia, Erb B2 phosphorylation (pErb B2), and epidermal nerve fiber density were assessed. The contribution of Erb B2 to DPN was assessed using the Erb B2 inhibitors PKI 166 and erlotinib and a conditional bitransgenic mouse that expressed a constitutively active form of Erb B2 in myelinated Schwann cells (SCs). RESULTS: Diabetic mice exhibited decreased MNCV and mechanical and thermal sensitivity, but the extent of these deficits was more severe in diabetic Cav1 knockout mice. Diabetes increased pErb B2 levels in both genotypes, but the absence of Cav1 correlated with a greater increase in pErb B2. Erb B2 activation contributed to the mechanical hypoalgesia and MNCV deficits in both diabetic genotypes because treatment with erlotinib or PKI 166 improved these indexes of DPN. Similarly, induction of a constitutively active Erb B2 in myelinated SCs was sufficient to decrease MNCV and induce a mechanical hypoalgesia in the absence of diabetes. CONCLUSIONS: Increased Erb B2 activity contributes to specific indexes of DPN, and Cav1 may be an endogenous regulator of Erb B2 signaling. Altered Erb B2 signaling is a novel mechanism that contributes to SC dysfunction in diabetes, and inhibiting Erb B2 may ameliorate deficits of tactile sensitivity in DPN. PMID- 19675141 TI - Time-modified confounding. AB - According to the authors, time-modified confounding occurs when the causal relation between a time-fixed or time-varying confounder and the treatment or outcome changes over time. A key difference between previously described time varying confounding and the proposed time-modified confounding is that, in the former, the values of the confounding variable change over time while, in the latter, the effects of the confounder change over time. Using marginal structural models, the authors propose an approach to account for time-modified confounding when the relation between the confounder and treatment is modified over time. An illustrative example and simulation show that, when time-modified confounding is present, a marginal structural model with inverse probability-of-treatment weights specified to account for time-modified confounding remains approximately unbiased with appropriate confidence limit coverage, while models that do not account for time-modified confounding are biased. Correct specification of the treatment model, including accounting for potential variation over time in confounding, is an important assumption of marginal structural models. When the effect of confounders on either the treatment or outcome changes over time, time modified confounding should be considered. PMID- 19675143 TI - Urban particulate matter causes ER stress and the unfolded protein response in human lung cells. AB - Because of its presumed adverse health effects, particulate air pollution (PM) has received growing attention, but the cellular mechanisms by which PM exerts toxicity are not well elucidated. PM has been associated with early mortality from illnesses that share endoplasmic reticulum (ER) stress as a mechanism of pathogenesis. In this study, we examined whether PM would induce the unfolded protein response (UPR) which is a cellular response to ER stress. Coarse (PM(10)) and fine (PM(2.5)) PM was collected from a single location in Northern Utah's Cache Valley during atmospheric inversions occurring in January 2002 and January 2003. Extracts of PM samples were added (12.5 and 25 microg/ml) to cultured human bronchial epithelial (BEAS-2B) cells for 24 h. At these concentrations neither PM nor LPS exhibited demonstrable cytotoxicity by the neutral red assay. However, PM elicited significant increases of unfolded protein response (UPR)-related post translational modifications, such as S6 ribosomal protein, heat-shock protein (Hsp)27, and protein kinase related protein phosphorylation and cleavage of activating transcription factor (ATF)-6. PM exposure also resulted in significant increases in the UPR-associated proteins ATF-4, Hsp70, Hsp90, and binding immunoglobulin protein. PM also interfered with the export of Hsp70 from the cells in a concentration-dependent manner and resulted in release of C-reactive protein. Calpain was upregulated and activated in PM-treated cultures, though these events were not proapoptotic. This study demonstrates that PM is capable of inducing ER stress and the UPR in vitro and may be a mechanism by which PM exerts toxicity. PMID- 19675144 TI - Automated dose-response analysis of the relative hepatic gene expression potency of TCDF in C57BL/6 mice. AB - Toxic equivalency factors (TEFs) are assigned to dioxin-like chemicals based on relative potency (REP) values of individual adaptive and toxic responses compared to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Agilent 4x44K oligonucleotide microarrays were used to examine the hepatic gene expression potency of 2,3,7,8 tetrachlorodibenzofuran (TCDF), relative to TCDD with complementary histopathology, TCDD and TCDF tissue level analysis, and ethoxyresorufin-O deethylase (EROD) assay data. Immature ovariectomized C57BL/6 mice were gavaged with 0.03, 0.1, 0.3, 1, 3, 10, 30, or 100 microg/kg TCDD, the World Health Organization TEF-adjusted doses (10 x TCDD dose) of TCDF (0.3, 1, 3, 10, 30, 100, or 300 microg/kg), or sesame oil vehicle and killed at 72 h. Two thousand two hundred eighty-eight and 1347 genes were differentially expressed (P1(t) > 0.90) at one or more doses by TCDD and TCDF, respectively. Automated dose-response modeling (ToxResponse Modeler) identified a total of 1027 and 837 genes with either a sigmoidal, exponential, linear, Gaussian, or quadratic dose-response relationship 72 h after treatment in TCDD and TCDF, respectively. Two hundred seventy genes exhibited a sigmoidal TCDD-induced dose-response (ED(50s) from 0.08 to 42.2 microg/kg) compared to only 179 sigmoidal responsive genes (ED(50s) from 0.74 to 299.9 microg/kg) elicited by TCDF. Of the 1027 TCDD dose-responsive genes, 654 were not examined further due to the lack of a dose response elicited by TCDF. Of the 373 genes that exhibited a TCDD and TCDF dose response, REPs were calculated for the 83 genes that exhibited comparable sigmoidal curve shapes and slopes. The median REP for these 83 genes was 0.10, with a maximum REP of 0.56 and a minimum of 0.01. REPs of 0.04 were also calculated for EROD and increase in relative liver weight (RLW) at 72 h. Collectively, the lower number of TCDF induced genes compared to TCDD and the 0.04 REPs for EROD activity and increased RLW are not consistent with the TEF of 0.10 for the hepatotoxicity of TCDF in C57BL/6 mice at 72 h. PMID- 19675142 TI - Gains in body fat and vasomotor symptom reporting over the menopausal transition: the study of women's health across the nation. AB - Although most women report vasomotor symptoms (hot flashes, night sweats) during midlife, their etiology and risk factors are incompletely understood. Body fat is positively associated with vasomotor symptoms cross-sectionally, but the longitudinal relation between changes in body fat and vasomotor symptoms is uncharacterized. The study aim was to examine whether gains in body fat were related to vasomotor symptom reporting over time. Measures of bioelectrical impedance for body fat, reproductive hormones, and reported vasomotor symptoms were assessed annually over 4 years from 2002 to 2006 among 1,659 women aged 47 59 years participating in the Study of Women's Health Across the Nation. Body fat change was examined in relation to vasomotor symptoms by using generalized estimating equations. Body fat gains were associated with greater odds of reporting hot flashes in models adjusted for age, site, race/ethnicity, education, smoking, parity, anxiety, and menopausal status (relative to stable body fat, gain: odds ratio = 1.23, 95% confidence interval: 1.02, 1.48; P = 0.03; loss: odds ratio = 1.07, 95% confidence interval: 0.89, 1.29; P = 0.45). Findings persisted controlling for estradiol, the free estradiol index, or follicle stimulating hormone concentrations. The relations between body fat changes and night sweats were not statistically significant. Body fat gains are associated with greater hot flash reporting during the menopausal transition. PMID- 19675145 TI - Simultaneous in vivo time course and dose response evaluation for TCDD-induced impairment of the LPS-stimulated primary IgM response. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a potent suppressor of humoral immunity but the specific molecular mechanisms responsible for immunosuppression by TCDD are poorly understood. In vivo and in vitro studies of the primary humoral IgM response demonstrated that the B cell is a sensitive cell type to modulation by TCDD. We hypothesized that in vivo administration of TCDD disrupts expression of transcription factors controlling B cell to plasma cell differentiation. Female C57BL6 mice were treated with a single dose of TCDD (3, 10, or 30 microg/kg) and/or vehicle (sesame oil). On day 4 post-TCDD administration mice were sensitized with 25 microg lipopolysacchride (LPS) by intraperitioneal injection to stimulate an immune response. Splenocytes were isolated on subsequent days following LPS, up to 3 days post-LPS, and the expression of IgM, XBP-1, PAX5, BCL-6, and Blimp-1 was assessed. TCDD treatment dose-dependently suppressed LPS-induced IgM antibody-forming cell number, which was correlated with decreased frequency of CD19+ CD138+ cells. Gene expression analysis revealed that TCDD caused a dose-dependent suppression of Igmicro chain, Igkappa chain, IgJ chain, XBP-1, and Blimp-1. TCDD also dose-dependently suppressed LPS-stimulated increases in Blimp-1 protein expression in CD19+ B cells. The deregulation of Blimp-1 expression by TCDD provides a partial explanation for the concomitant suppression of the IgM response and confirms previous observations established in vitro. PMID- 19675146 TI - Emerging approaches to broaden resistance of soybean to soybean cyst nematode as supported by gene expression studies. PMID- 19675147 TI - Post-genomics studies of developmental processes in legume seeds. PMID- 19675148 TI - Conservation of lotus and Arabidopsis basic helix-loop-helix proteins reveals new players in root hair development. AB - Basic helix-loop-helix (bHLH) proteins constitute a large family of transcriptional regulators in plants. Although they have been shown to play important roles in a wide variety of developmental processes, relatively few have been functionally characterized. Here, we describe the map-based cloning of the Lotus japonicus ROOTHAIRLESS1 (LjRHL1) locus. Deleterious mutations in this locus prevent root hair development, which also aborts root hair-dependent colonization of the host root by nitrogen-fixing bacteria. We show that the LjRHL1 gene encodes a presumed bHLH transcription factor that functions in a nonredundant manner to control root hair development in L. japonicus. Homology search and cross-species complementation experiments defined three members of the Arabidopsis (Arabidopsis thaliana) bHLH protein family, At2g24260, At4g30980, and At5g58010, as functionally equivalent to LjRHL1. Curiously, At2g24260 and At4g30980 mRNA species accumulate independently from the known positive regulators of root hair cell fate, while all three genes act in a partially redundant manner to regulate root hair development in Arabidopsis. PMID- 19675149 TI - Plastidial glyceraldehyde-3-phosphate dehydrogenase deficiency leads to altered root development and affects the sugar and amino acid balance in Arabidopsis. AB - Glycolysis is a central metabolic pathway that, in plants, occurs in both the cytosol and the plastids. The glycolytic glyceraldehyde-3-phosphate dehydrogenase (GAPDH) catalyzes the conversion of glyceraldehyde-3-phosphate to 1,3 bisphosphoglycerate with concomitant reduction of NAD(+) to NADH. Both cytosolic (GAPCs) and plastidial (GAPCps) GAPDH activities have been described. However, the in vivo functions of the plastidial isoforms remain unresolved. In this work, we have identified two Arabidopsis (Arabidopsis thaliana) chloroplast/plastid localized GAPDH isoforms (GAPCp1 and GAPCp2). gapcp double mutants display a drastic phenotype of arrested root development, dwarfism, and sterility. In spite of their low gene expression level as compared with other GAPDHs, GAPCp down regulation leads to altered gene expression and to drastic changes in the sugar and amino acid balance of the plant. We demonstrate that GAPCps are important for the synthesis of serine in roots. Serine supplementation to the growth medium rescues root developmental arrest and restores normal levels of carbohydrates and sugar biosynthetic activities in gapcp double mutants. We provide evidence that the phosphorylated pathway of Ser biosynthesis plays an important role in supplying serine to roots. Overall, these studies provide insights into the in vivo functions of the GAPCps in plants. Our results emphasize the importance of the plastidial glycolytic pathway, and specifically of GAPCps, in plant primary metabolism. PMID- 19675150 TI - Multiple antibiotic resistance in Arabidopsis is conferred by mutations in a chloroplast-localized transport protein. AB - Widespread antibiotic resistance is a major public health concern, and plants represent an emerging antibiotic exposure route. Recent studies indicate that crop plants fertilized with antibiotic-laden animal manure accumulate antibiotics; however, the molecular mechanisms of antibiotic entry and subcellular partitioning within plant cells remain unknown. Here, we report that mutations in the Arabidopsis (Arabidopsis thaliana) locus Multiple Antibiotic Resistance1 (MAR1) confer resistance, while MAR1 overexpression causes hypersensitivity to multiple aminoglycoside antibiotics. Additionally, yeast expressing MAR1 are hypersensitive to the aminoglycoside G418. MAR1 encodes a protein with 11 putative transmembrane domains with low similarity to ferroportin1 from Danio rerio. A MAR1:yellow fluorescent protein fusion localizes to the chloroplast, and chloroplasts from plants overexpressing MAR1 accumulate more of the aminoglycoside gentamicin, while mar1-1 mutant chloroplasts accumulate less than the wild type. MAR1 overexpression lines are slightly chlorotic, and chlorosis is rescued by exogenous iron. MAR1 expression is also down-regulated by low iron. These data suggest that MAR1 is a plastid transporter that is likely to be involved in cellular iron homeostasis and allows opportunistic entry of multiple antibiotics into the chloroplast. PMID- 19675151 TI - Computational finishing of large sequence contigs reveals interspersed nested repeats and gene islands in the rf1-associated region of maize. AB - The architecture of grass genomes varies on multiple levels. Large long terminal repeat retrotransposon clusters occupy significant portions of the intergenic regions, and islands of protein-encoding genes are interspersed among the repeat clusters. Hence, advanced assembly techniques are required to obtain completely finished genomes as well as to investigate gene and transposable element distributions. To characterize the organization and distribution of repeat clusters and gene islands across large grass genomes, we present 961- and 594-kb contiguous sequence contigs associated with the rf1 (for restorer of fertility1) locus in the near-centromeric region of maize (Zea mays) chromosome 3. We present two methods for computational finishing of highly repetitive bacterial artificial chromosome clones that have proved successful to close all sequence gaps caused by transposable element insertions. Sixteen repeat clusters were observed, ranging in length from 23 to 155 kb. These repeat clusters are almost exclusively long terminal repeat retrotransposons, of which the paleontology of insertion varies throughout the cluster. Gene islands contain from one to four predicted genes, resulting in a gene density of one gene per 16 kb in gene islands and one gene per 111 kb over the entire sequenced region. The two sequence contigs, when compared with the rice (Oryza sativa) and sorghum (Sorghum bicolor) genomes, retain gene colinearity of 50% and 71%, respectively, and 70% and 100%, respectively, for high-confidence gene models. Collinear genes on single gene islands show that while most expansion of the maize genome has occurred in the repeat clusters, gene islands are not immune and have experienced growth in both intragene and intergene locations. PMID- 19675152 TI - In folio respiratory fluxomics revealed by 13C isotopic labeling and H/D isotope effects highlight the noncyclic nature of the tricarboxylic acid "cycle" in illuminated leaves. AB - While the possible importance of the tricarboxylic acid (TCA) cycle reactions for leaf photosynthesis operation has been recognized, many uncertainties remain on whether TCA cycle biochemistry is similar in the light compared with the dark. It is widely accepted that leaf day respiration and the metabolic commitment to TCA decarboxylation are down-regulated in illuminated leaves. However, the metabolic basis (i.e. the limiting steps involved in such a down-regulation) is not well known. Here, we investigated the in vivo metabolic fluxes of individual reactions of the TCA cycle by developing two isotopic methods, (13)C tracing and fluxomics and the use of H/D isotope effects, with Xanthium strumarium leaves. We provide evidence that the TCA "cycle" does not work in the forward direction like a proper cycle but, rather, operates in both the reverse and forward directions to produce fumarate and glutamate, respectively. Such a functional division of the cycle plausibly reflects the compromise between two contrasted forces: (1) the feedback inhibition by NADH and ATP on TCA enzymes in the light, and (2) the need to provide pH-buffering organic acids and carbon skeletons for nitrate absorption and assimilation. PMID- 19675153 TI - Remodeled respiration in ndufs4 with low phosphorylation efficiency suppresses Arabidopsis germination and growth and alters control of metabolism at night. AB - Respiratory oxidative phosphorylation is a cornerstone of cellular metabolism in aerobic multicellular organisms. The efficiency of this process is generally assumed to be maximized, but the presence of dynamically regulated nonphosphorylating bypasses implies that plants can alter phosphorylation efficiency and can benefit from lowered energy generation during respiration under certain conditions. We characterized an Arabidopsis (Arabidopsis thaliana) mutant, ndufs4 (for NADH dehydrogenase [ubiquinone] fragment S subunit 4), lacking complex I of the respiratory chain, which has constitutively lowered phosphorylation efficiency. Through analysis of the changes to mitochondrial function as well as whole cell transcripts and metabolites, we provide insights into how cellular metabolism flexibly adapts to reduced phosphorylation efficiency and why this state may benefit the plant by providing moderate stress tolerance. We show that removal of the single protein subunit NDUFS4 prevents assembly of complex I and removes its function from mitochondria without pleiotropic effects on other respiratory components. However, the lack of complex I promotes broad changes in the nuclear transcriptome governing growth and photosynthetic function. We observed increases in organic acid and amino acid pools in the mutant, especially at night, concomitant with alteration of the adenylate content. While germination is delayed, this can be rescued by application of gibberellic acid, and root growth assays of seedlings show enhanced tolerance to cold, mild salt, and osmotic stress. We discuss these observations in the light of recent data on the knockout of nonphosphorylating respiratory bypass enzymes that show opposite changes in metabolites and stress sensitivity. Our data suggest that the absence of complex I alters the adenylate control of cellular metabolism. PMID- 19675154 TI - Quantitative proteomics of seed filling in castor: comparison with soybean and rapeseed reveals differences between photosynthetic and nonphotosynthetic seed metabolism. AB - Seed maturation or seed filling is a phase of development that plays a major role in the storage reserve composition of a seed. In many plant seeds photosynthesis plays a major role in this process, although oilseeds, such as castor (Ricinus communis), are capable of accumulating oil without the benefit of photophosphorylation to augment energy demands. To characterize seed filling in castor, a systematic quantitative proteomics study was performed. Two-dimensional gel electrophoresis was used to resolve and quantify Cy-dye-labeled proteins expressed at 2, 3, 4, 5, and 6 weeks after flowering in biological triplicate. Expression profiles for 660 protein spot groups were established, and of these, 522 proteins were confidently identified by liquid chromatography-tandem mass spectrometry by mining against the castor genome. Identified proteins were classified according to function, and the most abundant groups of proteins were involved in protein destination and storage (34%), energy (19%), and metabolism (15%). Carbon assimilatory pathways in castor were compared with previous studies of photosynthetic oilseeds, soybean (Glycine max) and rapeseed (Brassica napus). These comparisons revealed differences in abundance and number of protein isoforms at numerous steps in glycolysis. One such difference was the number of enolase isoforms and their sum abundance; castor had approximately six times as many isoforms as soy and rapeseed. Furthermore, Rubisco was 11-fold less prominent in castor compared to rapeseed. These and other differences suggest some aspects of carbon flow, carbon recapture, as well as ATP and NADPH production in castor differs from photosynthetic oilseeds. PMID- 19675155 TI - Plant-derived sucrose is a key element in the symbiotic association between Trichoderma virens and maize plants. AB - Fungal species belonging to the genus Trichoderma colonize the rhizosphere of many plants, resulting in beneficial effects such as increased resistance to pathogens and greater yield and productivity. However, the molecular mechanisms that govern the recognition and association between Trichoderma and their hosts are still largely unknown. In this report, we demonstrate that plant-derived sucrose (Suc) is an important resource provided to Trichoderma cells and is also associated with the control of root colonization. We describe the identification and characterization of an intracellular invertase from Trichoderma virens (TvInv) important for the mechanisms that control the symbiotic association and fungal growth in the presence of Suc. Gene expression studies revealed that the hydrolysis of plant-derived Suc in T. virens is necessary for the up-regulation of Sm1, the Trichoderma-secreted elicitor that systemically activates the defense mechanisms in leaves. We determined that as a result of colonization of maize (Zea mays) roots by T. virens, photosynthetic rate increases in leaves and the functional expression of tvinv is crucial for such effect. In agreement, the steady-state levels of mRNA for Rubisco small subunit and the oxygen-evolving enhancer 3-1 were increased in leaves of plants colonized by wild-type T. virens. We conclude that during the symbiosis, the sucrolytic activity in the fungal cells affects the sink activity of roots, directing carbon partitioning toward roots and increasing the rate of photosynthesis in leaves. A discussion of the role of Suc in controlling the fungal proliferation on roots and its pivotal role in the coordination of plant-microbe associations is provided. PMID- 19675157 TI - Analysis of PHOTOPERIOD SENSITIVITY5 sheds light on the role of phytochromes in photoperiodic flowering in rice. AB - A great number of plants synchronize flowering with day length. In rice (Oryza sativa), photoperiod is the primary environmental cue that triggers flowering. Here, we show that the s73 mutant, identified in a gamma-irradiated Bahia collection, displays early flowering and photoperiodic insensitivity due to a null mutation in the PHOTOPERIOD SENSITIVITY5 (SE5) gene, which encodes an enzyme implicated in phytochrome chromophore biosynthesis. s73 mutant plants show a number of alterations in the characteristic diurnal expression patterns of master genes involved in photoperiodic control of flowering, resulting in up-regulation of the floral integrator Heading date3a (Hd3a). Early heading date1 (Ehd1), an additional rice floral activator, was also highly expressed in the s73 mutant, suggesting that SE5 represses Ehd1 in wild-type plants. Silencing of Ehd1 in both Bahia and s73 backgrounds indicated that SE5 regulates Ehd1 expression. The data also indicate that SE5 confers photoperiodic sensitivity through regulation of Hd1. These results provide direct evidence that phytochromes inhibit flowering by affecting both Hd1 and Ehd1 flowering pathways. PMID- 19675156 TI - Unraveling the evolution of cytokinin signaling. AB - The conquest of the land by plants required dramatic morphological and metabolic adaptations. Complex developmental programs under tight regulation evolved during this process. Key regulators of plant development are phytohormones, such as cytokinins. Cytokinins are adenine derivatives that affect various processes in plants. The cytokinin signal transduction system, which is mediated via a multistep variant of the bacterial two-component signaling system, is well characterized in the model plant Arabidopsis (Arabidopsis thaliana). To understand the origin and evolutionary pattern of this signaling pathway, we surveyed the genomes of several sequenced key plant species ranging from unicellular algae, moss, and lycophytes, to higher land plants, including Arabidopsis and rice (Oryza sativa), for proteins involved in cytokinin signal transduction. Phylogenetic analysis revealed that the hormone-binding receptor and a class of negative regulators first appeared in land plants. Other components of the signaling pathway were present in all species investigated. Furthermore, we found that the receptors evolved under different evolutionary constraints from the other components of the pathway: The number of receptors remained fairly constant, while the other protein families expanded. PMID- 19675158 TI - CCR7 deficiency in NOD mice leads to thyroiditis and primary hypothyroidism. AB - CCR7 is involved in the initiation of immune responses and has been recently implicated in the control of tolerance. To analyze the role of CCR7 in autoimmunity, we backcrossed CCR7(ko/ko) mice (in which ko signifies deficient) onto the autoimmune-prone NOD background. Surprisingly, NODCCR7(ko/ko) mice never developed diabetes, but showed severe inflammation in multiple tissues including thyroid, lung, stomach, intestine, uterus, and testis. NODCCR7(ko/ko) mice had a marked enlargement of the thyroid gland (goiter) that was associated with circulating autoantibodies against thyroglobulin, and development of primary hypothyroidism (decreased levels of serum thyroxin, and augmented levels of thyroid-stimulating hormone in the pituitary gland), features found in Hashimoto's thyroiditis. Cells isolated from diseased thyroids and activated splenocytes from NODCCR7(ko/ko) animals induced goiter in NOD.SCID recipients, demonstrating that autoreactive cells were generated in the absence of CCR7. Moreover, thyroid disease could be accelerated in young NODCCR7(ko/ko) mice by immunization with thyroglobulin. These results demonstrate the complexity in the generation of multiple autoimmune phenotypes in NOD mice and indicate that CCR7 is a key molecule in their development. PMID- 19675159 TI - Crucial contribution of thymic Sirp alpha+ conventional dendritic cells to central tolerance against blood-borne antigens in a CCR2-dependent manner. AB - Thymic dendritic cells (DCs) as well as thymic epithelial cells are presumed to be major sentinels in central tolerance by inducing the apoptosis of autoreactive T progenitor cells. The thymic DC population is composed of heterogeneous subsets including CD11c(+)B220(+) plasmacytoid DCs, CD11c(+)B220(-)CD8alpha(+) signal regulatory protein alpha (Sirpalpha)(-) and CD11c(+)B220(-)CD8alpha( )Sirpalpha(+) conventional DCs (cDCs). However, the distinctive role of each DC subset remains undefined. We show herein that Sirpalpha(+) cDCs, a minor subpopulation, was disseminated in the thymic cortical area with some of them uniquely localized inside perivascular regions and nearby small vessels in the thymus. The Sirpalpha(+) but not Sirpalpha(-) cDC subset can selectively capture blood-circulating Ags. Moreover, in CCR2-deficient mice, the thymic Sirpalpha(+) cDC subset, but not other thymic cell components, was moderately decreased especially in the perivascular regions. Concomitantly, these mice exhibited a modest impairment in intrathymic negative selection against blood-borne Ags, with the reduced capacity to uptake blood-borne Ags. Given their intrathymic cortical localization, CD11c(+)B220(-)CD8alpha(-)Sirpalpha(+) cDCs can have a unique role in the development of central tolerance against circulating peripheral Ags, at least partially in a CCR2-dependent manner. PMID- 19675160 TI - Genetic control of severe egg-induced immunopathology and IL-17 production in murine schistosomiasis. AB - Infection with the trematode parasite Schistosoma mansoni results in a distinct heterogeneity of disease severity, both in humans and in an experimental mouse model. Severe disease is characterized by pronounced hepatic egg-induced granulomatous inflammation in a proinflammatory cytokine environment, whereas mild disease corresponds with reduced hepatic inflammation in a Th2 skewed cytokine environment. This marked heterogeneity indicates that genetic differences play a significant role in disease development, yet little is known about the genetic basis of dissimilar immunopathology. To investigate the role of genetic susceptibility in murine schistosomiasis, quantitative trait loci analysis was performed on F(2) progeny derived from SJL/J and C57BL/6 mice, which develop severe and mild pathology, respectively. In this study, we show that severe liver pathology in F(2) mice 7 wk after infection significantly correlated with an increase in the production of the proinflammatory cytokines IL-17, IFN gamma, and TNF-alpha by schistosome egg Ag-stimulated mesenteric lymph node cells. Quantitative trait loci analysis identified several genetic intervals controlling immunopathology as well as IL-17 and IFN-gamma production. Egg granuloma size exhibited significant linkage to two loci, D4Mit203 and D17Mit82, both of which were inherited in a BL/6 dominant manner. Furthermore, a significant reduction of hepatic granulomatous inflammation and IL-17 production in interval-specific congenic mice demonstrated that the two identified genetic loci have a decisive effect on the development of immunopathology in murine schistosomiasis. PMID- 19675161 TI - CXCR2-dependent mucosal neutrophil influx protects against colitis-associated diarrhea caused by an attaching/effacing lesion-forming bacterial pathogen. AB - Enteropathogenic Escherichia coli (EPEC) is a major cause of diarrheal disease in young children, yet symptoms and duration are highly variable for unknown reasons. Citrobacter rodentium, a murine model pathogen that shares important functional features with EPEC, colonizes mice in colon and cecum and causes inflammation, but typically little or no diarrhea. We conducted genome-wide microarray studies to define mechanisms of host defense and disease in C. rodentium infection. A significant fraction of the genes most highly induced in the colon by infection encoded CXC chemokines, particularly CXCL1/2/5 and CXCL9/10, which are ligands for the chemokine receptors CXCR2 and CXCR3, respectively. CD11b(+) dendritic cells were the major producers of CXCL1, CXCL5, and CXCL9, while CXCL2 was mainly induced in macrophages. Infection of gene targeted mice revealed that CXCR3 had a significant but modest role in defense against C. rodentium, whereas CXCR2 had a major and indispensable function. CXCR2 was required for normal mucosal influx of neutrophils, which act as direct antibacterial effectors. Moreover, CXCR2 loss led to severe diarrhea and failure to express critical components of normal ion and fluid transport, including ATPase beta(2)-subunit, CFTR, and DRA. The antidiarrheal functions were unique to CXCR2, since other immune defects leading to increased bacterial load and inflammation did not cause diarrhea. Thus, CXCR2-dependent processes, particularly mucosal neutrophil influx, not only contribute to host defense against C. rodentium, but provide protection against infection-associated diarrhea. PMID- 19675162 TI - Compound A, a plant origin ligand of glucocorticoid receptors, increases regulatory T cells and M2 macrophages to attenuate experimental autoimmune neuritis with reduced side effects. AB - Experimental autoimmune neuritis (EAN) is a helper T cell-mediated autoimmune demyelinating inflammatory disease of the peripheral nervous system and serves as the animal model for human inflammatory demyelinating polyneuropathies. Compound A, a plant-derived phenyl aziridine precursor, was reported to activate glucocorticoid receptors to exert transrepression but not transactivation properties. In this study, we investigated the effects of Compound A in EAN rats. Compound A greatly suppressed paraparesis in EAN, even when administrated after the appearance of the first neurological signs. Accumulation of macrophages and lymphocytes, demyelination, and mRNA levels of inflammatory molecules in sciatic nerves of EAN were greatly attenuated by Compound A. In addition, Compound A inhibited progression of neuropathic pain and repressed microglia but not astrocyte activation and IL-1beta and TNF-alpha up-regulation in EAN spinal cords. In EAN sciatic nerves, Compound A treatment increased numbers of anti inflammatory M2 macrophages. Furthermore, Compound A induced the switch of macrophages from inflammatory M1 type to anti-inflammatory M2 type in vitro. In lymph nodes of EAN rats, Compound A depressed Th1 and Th17 cytokines, but increased Th2 cytokine and Foxp3 expression. An increase of Foxp3(+)/CD4(+) regulatory T cells was seen in peripheral blood of EAN rats following Compound A treatment. In addition, Compound A did not cause a hyperglycemia effect in EAN rats as compared with the immunosuppressive steroid prednisolone. Therefore, our data demonstrated that Compound A could effectively suppress EAN with reduced side effects by attenuating inflammation, suggesting that Compound A could be a potent candidate for treatment of autoimmune neuropathies. PMID- 19675164 TI - Ornithorhynchus anatinus (platypus) links the evolution of immunoglobulin genes in eutherian mammals and nonmammalian tetrapods. AB - The evolutionary origins of mammalian immunoglobulin H chain isotypes (IgM, IgD, IgG, IgE, and IgA) are still incompletely understood as these isotypes differ considerably in structure and number from their counterparts in nonmammalian tetrapods. We report in this study that the platypus (Ornithorhynchus anatinus) Ig H chain constant region gene locus contains eight Ig encoding genes, which are arranged in an mu-delta-omicron-gamma2-gamma1-alpha1-epsilon-alpha2 order, spanning a total of approximately 200 kb DNA, encoding six distinct isotypes. The omicron (omicron for Ornithorhynchus) gene encodes a novel Ig H chain isotype that consists of four constant region domains and a hinge, and is structurally different from any of the five known mammalian Ig classes. This gene is phylogenetically related to upsilon (epsilon) and gamma, and thus appears to be a structural intermediate between these two genes. The platypus delta gene encodes ten heavy chain constant region domains, lacks a hinge region and is similar to IgD in amphibians and fish, but strikingly different from that in eutherian mammals. The platypus Ig H chain isotype repertoire thus shows a unique combination of genes that share similarity both to those of nonmammalian tetrapods and eutherian animals and demonstrates how phylogenetically informative species can be used to reconstruct the evolutionary history of functionally important genes. PMID- 19675163 TI - Memory-like CD8+ T cells generated during homeostatic proliferation defer to antigen-experienced memory cells. AB - Naive T cells proliferate in response to lymphopenia and acquire the phenotypic and functional qualities of memory T cells, providing enhanced protection against infection. How well memory-like T cells generated during lymphopenia-induced homeostatic proliferation (HP)-memory differentiate into secondary memory cells and compete with Ag-experienced true-memory cells is unknown. We found that CD8(+) HP-memory T cells generated robust responses upon infection and produced a secondary memory population comparable to true-memory cells in the absence of competition. However, when true-memory and HP-memory T cells competed during infection, HP-memory cells contributed less to the effector population, contracted earlier, and formed fewer secondary memory cells. Furthermore, HP- and true-memory cells demonstrated distinct chemokine receptor expression and localization within the spleen during infection, indicating differential access to signals necessary for secondary memory formation. Thus, HP-memory T cells provide protection without compromising the true-memory population. Differences in HP- and true-memory T cells may reveal the basis of competition for limited resources within the memory-T cell compartment. PMID- 19675165 TI - Phosphorylation of Nur77 by the MEK-ERK-RSK cascade induces mitochondrial translocation and apoptosis in T cells. AB - Nur77, an orphan nuclear receptor, plays a key role in apoptosis in T cells. In cancer cell lines, Nur77 can induce apoptosis through the intrinsic apoptotic pathway, but the mechanism by which Nur77 kills T cells remains controversial. In this study, we provide biochemical, pharmacological, and genetic evidence demonstrating that Nur77 induces apoptosis through the activation of the intrinsic pathway in T cells. We also show that Nur77 is a physiological substrate of the MEK-ERK-RSK cascade. Specifically, we demonstrate that RSK phosphorylates Nur77 at serine 354 and this modulates Nur77 nuclear export and intracellular translocation during T cell death. Our data reveal that Nur77 phosphorylation and mitochondrial targeting, regulated by RSK, defines a role for the MEK1/2-ERK1/2 cascade in T cell apoptosis. PMID- 19675166 TI - Stimulatory and inhibitory killer Ig-like receptor molecules are expressed and functional on lupus T cells. AB - T cells from lupus patients have hypomethylated DNA and overexpress genes normally suppressed by DNA methylation that contribute to disease pathogenesis. We found that stimulatory and inhibitory killer cell Ig-like receptor (KIR) genes are aberrantly overexpressed on experimentally demethylated T cells. We therefore asked if lupus T cells also overexpress KIR, and if the proteins are functional. T cells from lupus patients were found to overexpress KIR genes, and expression was proportional to disease activity. Abs to the stimulatory molecule KIR2DL4 triggered IFN-gamma release by lupus T cells, and production was proportional to disease activity. Similarly, cross-linking the inhibitory molecule KIR3DL1 prevented the autoreactive macrophage killing that characterizes lupus T cells. These results indicate that aberrant T cell KIR expression may contribute to IFN overproduction and macrophage killing in human lupus, and they suggest that Abs to inhibitory KIR may be a treatment for this disease. PMID- 19675167 TI - Self-peptides prolong survival in murine autoimmunity via reduced IL-2/IL-7 mediated STAT5 signaling, CD8 coreceptor, and V alpha 2 down-regulation. AB - Although the pathogenic role of B cells and CD4 T cells has been studied extensively, less is known about the role of CD8 T cells in autoimmunity and self tolerance. To evaluate the role of CD8 T cells in autoimmunity and its modulation using self-peptides, we used mice expressing soluble OVA (sOVA) under control of the keratin-14 promoter. Spontaneous autoimmunity occurred when sOVA mice were crossed with OT-I mice, whose CD8 T cells carry a Valpha2/Vbeta5-transgenic TCR with specificity for the OVA(257-264) peptide. Eighty-three percent of OVA/OT-I mice died during the first 2 wk of life due to multiple organ inflammation. In contrast, preventive or therapeutic OVA(257-264) peptide injections induced a dose-dependent increase in survival. Healthy survivors exhibited reductions in peripheral CD8 T cells, CD8 coreceptor, and Valpha2 expression. Furthermore, CD8 T cells from healthy mice were anergic and could not be activated by exogenous IL 2. A block in IL-2/IL-7 signaling via the STAT5 pathway provided the basis for low surface expression of the CD8 coreceptor and failure of IL-2 to break CD8 T cell anergy. Thus, the soluble TCR ligand triggered multiple tolerance mechanisms in these sOVA/OT-I mice, making this treatment approach a potential paradigm for modulating human autoimmune diseases. PMID- 19675168 TI - Sequential, ordered acquisition of antibodies to Plasmodium falciparum erythrocyte membrane protein 1 domains. AB - The binding of erythrocytes infected with mature blood stage parasites to the vascular bed is key to the pathogenesis of malignant malaria. The binding is mediated by members of Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) family. PfEMP1s can be divided into groups, and it has previously been suggested that parasites expressing group A or B/A PfEMP1s are most pathogenic. To test the hypothesis that the first malaria infections in infants and young children are dominated by parasites expressing A and B/A PfEMP1s, we measured the plasma Ab level against 48 recombinant PfEMP1 domains of different groupings in 1342 individuals living in five African villages characterized by markedly different malaria transmission. We show that children progressively acquire a broader repertoire of anti-PfEMP1 Abs, but that the rate of expansion is governed by transmission intensity. However, independently of transmission intensity, Abs are first acquired to particular Duffy binding ligand-like domains belonging to group A or B/A PfEMP1s. The results support the view that anti-PfEMP1 Ab responses effectively structure the expenditure of the repertoire of PfEMP1 maintained by the parasite. Parasites expressing certain group A and B/A PfEMP1s are responded to first by individuals with limited previous exposure, and the resulting Abs reduce the fitness and pathogenicity of these parasites during subsequent infections. This allows parasites expressing less pathogenic PFEMP1s to dominate during later infections. The identification of PfEMP1 domains expressed by parasites causing disease in infants and young children is important for development of vaccines protecting against severe malaria. PMID- 19675170 TI - IL-10 is a negative regulatory factor of CAWS-vasculitis in CBA/J mice as assessed by comparison with Bruton's tyrosine kinase-deficient CBA/N mice. AB - Candida albicans water-soluble fraction (CAWS), a mannoprotein-beta-glucan complex obtained from the culture supernatant of C. albicans NBRC1385, exhibits vasculitis-inducing activity (CAWS-vasculitis) in mice. The sensitivity to CAWS vasculitis varies greatly among mouse strains. This study examined the factors contributing to or inhibiting CAWS-vasculitis using CAWS-vasculitis-resistant CBA/J mice and Bruton's tyrosine kinase-deficient CBA/N mice, which is a CAWS vasculitis-sensitive strain that has the same origin as CBA/J mice. After stimulation with various kinds of pathogen-associated molecular patterns, the production of inflammatory cytokines IL-6 and IFN-gamma was induced in CBA/N mice, whereas that of immunosuppressive IL-10 was induced in CAWS-vasculitis resistant CBA/J mice. Furthermore, the production of tissue inhibitor of metalloproteinase 1, an endogenous matrix metalloproteinase inhibitor, was observed in CBA/J mice. The results strongly suggest that the difference in the production of these cytokines is closely linked to the development of CAWS vasculitis. PMID- 19675169 TI - Processing and presentation of variant surface glycoprotein molecules to T cells in African trypanosomiasis. AB - Th1 cell responses to the variant surface glycoprotein (VSG) of African trypanosomes play a critical role in controlling infection through the production of IFN-gamma, but the role of APCs in the induction and regulation of T cell mediated protection is poorly understood. In this study, we have investigated the Ag presentation capabilities of dendritic cells (DCs) and macrophages during early trypanosome infection in relatively resistant responder and susceptible nonresponder mouse strains. Splenic DCs appeared to be the primary cell responsible for activating naive VSG-specific Th cell responses in resistant responder animals through the coordinated up-regulation of costimulatory molecules, secretion of IL-12, and presentation of VSG peptides to T cells in vivo. Splenic DC depletion and the down-regulation of costimulatory markers on splenic macrophages were observed in susceptible animals and may be associated with the inability of these animals to elicit a significant VSG-specific T cell response. In contrast to splenic APCs, peritoneal macrophages secreted NO, failed to activate naive Th cells in vitro, and presented relatively low levels of VSG peptides to T cells in vivo. Thus, VSG-specific Th1 cell responses may be determined by tissue- and cell-specific differences in Ag presentation. Additionally, all APCs from resistant and susceptible strains displayed a reduced ability to process and present newly encountered exogenous Ag, including new VSG molecules, during high parasitemia. Thus, initial uptake of VSG (or other trypanosome factors) may interfere with Ag presentation and have dramatic consequences for subsequent T cell responses to other proteins. PMID- 19675171 TI - HLA-DR alleles in amyloid beta-peptide autoimmunity: a highly immunogenic role for the DRB1*1501 allele. AB - Active amyloid beta-peptide (Abeta) immunization of patients with Alzheimer's disease (AD) caused meningoencephalitis in approximately 6% of immunized patients in a clinical trial. In addition, long-term studies of AD patients show varying degrees of Abeta Ab responses, which correlate with the extent of Abeta clearance from the brain. In this study, we examined the contribution of various HLA-DR alleles to these immune-response variations by assessing Abeta T cell reactivity, epitope specificity, and immunogenicity. Analysis of blood samples from 133 individuals disclosed that the abundant DR haplotypes DR15 (found in 36% of subjects), DR3 (in 18%), DR4 (12.5%), DR1 (11%), and DR13 (8%) were associated with Abeta-specific T cell responses elicited via distinct T cell epitopes within residues 15-42 of Abeta. Because the HLA-DRB1*1501 occurred most frequently, we examined the effect of Abeta challenge in humanized mice bearing this allele. The observed T cell response was remarkably strong, dominated by secretion of IFN gamma and IL-17, and specific to the same T cell epitope as that observed in the HLA-DR15-bearing humans. Furthermore, following long-term therapeutic immunization of an AD mouse model bearing the DRB1*1501 allele, Abeta was effectively cleared from the brain parenchyma and brain microglial activation was reduced. The present study thus characterizes HLA-DR alleles directly associated with specific Abeta T cell epitopes and demonstrates the highly immunogenic properties of the abundant allele DRB1*1501 in a mouse model of AD. This new knowledge enables us to explore the basis for understanding the variations in naturally occurring Abeta-reactive T cells and Abeta immunogenicity among humans. PMID- 19675173 TI - 12/15-lipoxygenase counteracts inflammation and tissue damage in arthritis. AB - Eicosanoids are essential mediators of the inflammatory response and contribute both to the initiation and the resolution of inflammation. Leukocyte-type 12/15 lipoxygenase (12/15-LO) represents a major enzyme involved in the generation of a subclass of eicosanoids, including the anti-inflammatory lipoxin A(4) (LXA(4)). Nevertheless, the impact of 12/15-LO on chronic inflammatory diseases such as arthritis has remained elusive. By using two experimental models of arthritis, the K/BxN serum-transfer and a TNF transgenic mouse model, we show that deletion of 12/15-LO leads to uncontrolled inflammation and tissue damage. Consistent with these findings, 12/15-LO-deficient mice showed enhanced inflammatory gene expression and decreased levels of LXA(4) within their inflamed synovia. In isolated macrophages, the addition of 12/15-LO-derived eicosanoids blocked both phosphorylation of p38MAPK and expression of a subset of proinflammatory genes. Conversely, 12/15-LO-deficient macrophages displayed significantly reduced levels of LXA(4), which correlated with increased activation of p38MAPK and an enhanced inflammatory gene expression after stimulation with TNF-alpha. Taken together, these results support an anti-inflammatory and tissue-protective role of 12/15-LO and its products during chronic inflammatory disorders such as arthritis. PMID- 19675174 TI - Benefits of negative social exchanges for emotional closeness. AB - Negative exchanges in social relationships have traditionally been studied as having negative consequences. This study explored whether they might have positive effects for relationship closeness. The sample included 351 adults, aged between 18 and 91 years, residing in Hong Kong, China. Closeness of social partners to the participants was measured by the Social Convoy Questionnaire, and the levels of negative exchanges and social support from each social partner were assessed. Multilevel analyses revealed that more negative exchanges were associated with a more positive change in closeness over a 2-year period, even after statistically controlling for social support and sociostructural characteristics of the participant and the social partner. Findings extended our knowledge on the positive effects of negative exchanges and their moderating conditions. PMID- 19675172 TI - Modulation of single-cell IgG secretion frequency and rates in human memory B cells by CpG DNA, CD40L, IL-21, and cell division. AB - During the recall response by CD27(+) IgG class-switched human memory B cells, total IgG secreted is a function of the following: 1) the number of IgG-secreting cells (IgG-SC), and 2) the secretion rate of each cell. In this study, we report the quantitative ELISPOT method for simultaneous estimation of single-cell IgG secretion rates and secreting cell frequencies in human B cell populations. We found that CD27(+) IgM(-) memory B cells activated with CpG and cytokines had considerable heterogeneity in the IgG secretion rates, with two major secretion rate subpopulations. BCR cross-linking reduced the frequency of cells with high per-cell IgG secretion rates, with a parallel decrease in CD27(high) B cell blasts. Increased cell death may account for the BCR-stimulated reduction in high rate IgG-SC CD27(high) B cell blasts. In contrast, the addition of IL-21 to CD40L plus IL-4-activated human memory B cells induced a high-rate IgG-SC population in B cells with otherwise low per-cell IgG secretion rates. The profiles of human B cell IgG secretion rates followed the same biphasic distribution and range irrespective of division class. This, along with the presence of non-IgG producing, dividing B cells in CpG plus cytokine-activated B memory B cell populations, is suggestive of an on/off switch regulating IgG secretion. Finally, these data support a mixture model of IgG secretion in which IgG secreted over time is modulated by the frequency of IgG-SC and the distribution of their IgG secretion rates. PMID- 19675175 TI - Ploidy-mediated reduced segregation facilitates fixation of heterozygosity in the aromatic grass, Cymbopogon martinii (Roxb.). AB - In most medicinal and aromatic plants, the vegetative tissue (e.g., roots, stems, leaves) is the source of the economic product. These plants are inherently heterozygous (natural allelic hybrids) and maintain their genetic makeup in nature by obligate vegetative propagation. Under seed cultivation, these plants incur population heterogeneity that reduces biomass and hampers product quality. Therefore, fixation of heterozygosity is vital for maintaining uniformity in quality of the economic product and quantity of biomass under seed cultivation. Although seed-grown clonal progenies identical to the mother plant can be obtained in certain plants that show an unusual breeding system called apomixis, such a breeding system is rare in medicinal and aromatic plants of economic value. Here we show an effective experimental strategy based on a polyploid model that facilitates fixation of heterozygosity in obligate asexual species owing to tetrasomic inheritance and low segregation in C(1) progenies from high-fertility C(0) autopolyploids. Using an obligate asexual species of aromatic grass Cymbopogon martinii, we demonstrated that progenitor diploids with distal chiasma localization and low chiasmate association in meiosis, when changed into tetraploids, entail high gametic/seed fertility reflected in high bivalent pairing and balanced anaphase segregation. Their seed progenies evince crop homogeneity owing to reduced segregation, indicating fixation of heterozygosity present in the source diploids. Because C. martinii could be maintained through obligate vegetative propagation, here is a unique opportunity to utilize the polyploid advantage through C(1) seed progenies for commercial cultivation, as well as maintenance of original C(0) stock for raising seeds without losing polyploid heterosis normally threatened in subsequent segregating progenies on account of aneuploidy and gametic instability. PMID- 19675176 TI - Religious attendance reduces cognitive decline among older women with high levels of depressive symptoms. AB - BACKGROUND: There is growing evidence that regular attendance at religious functions is associated with less cognitive decline (CD). However, little research has investigated factors that may moderate the religious attendance-CD relationship. The present study examined the effects of gender and depressive symptoms on the relationship between religious attendance and CD. METHODS: Data were drawn from waves 1 and 2 of the Duke Established Populations for Epidemiologic Studies of the Elderly, which were 3 years apart. Participants consisted of a sample of community-dwelling older adults aged 65 years and older (N = 2,938). Linear regression analyses were conducted controlling for important demographic-, socioeconomic-, and health-related variables. Cognitive functioning was assessed at both waves to examine change in errors over time. RESULTS: Greater religious attendance was related to less CD. In addition, there was a three-way interaction between religious attendance, gender, and depressive symptoms in predicting CD. Among women with higher levels of depressive symptoms, those who less frequently attended religious services experienced greater CD than those who more frequently attended religious services. The interaction between attendance and depressive symptoms in men did not reach significance. CONCLUSIONS: Religious attendance may offer mental stimulation that helps to maintain cognitive functioning in later life, particularly among older depressed women. Given the possible benefits religious attendance may have on cognitive functioning, it may be appropriate in certain instances for clinicians to recommend that clients reengage in religious activities they may have given up as a result of their depression. PMID- 19675177 TI - Driving cessation and health trajectories in older adults. AB - BACKGROUND: Older adults who cease driving have poorer health than those who continue to drive. However, it is unclear whether the transition to driving cessation itself results in health declines or whether driving cessation subsequently exacerbates health declines over time. METHODS: The current study addresses these questions using multilevel modeling among 690 older adults from the Advanced Cognitive Training for Independent and Vital Elderly study. Driving status and health, as indicated by the SF-36 questionnaire, self-rated health, physical performance (Turn 360 degrees Test), and depressive symptoms were assessed at baseline and at 1-, 2-, 3-, and 5-year follow-up visits. RESULTS: The transition to driving cessation was accompanied by significant declines in physical and social functioning, physical performance, and physical role (ps < .05). Health declines after driving cessation were steeper for general health. CONCLUSIONS: The transition to driving cessation is associated with health declines for older adults as measured by several indicators. Additionally, general health declines more sharply following driving cessation. These findings highlight the importance of interventions to sustain driving mobility among older adults. PMID- 19675179 TI - Toll-like receptor 4 (TLR-4) regulates renal ion transport. PMID- 19675178 TI - Early exposure to germs modifies kidney damage and inflammation after experimental ischemia-reperfusion injury. AB - Kidney ischemia-reperfusion injury (IRI) is, in part, mediated by immune and inflammatory factors. Since microbial stimuli are known to alter immune and inflammatory responses, we hypothesized that differences in perinatal microbial status would modify renal injury following IRI. We performed bilateral renal IRI on 6-wk-old germ-free and control mice and studied the effects on kidney lymphocyte trafficking, cytokines, function, and structure. Compared with control mice, normal kidneys of germ-free mice exhibited more NKT cells and lower IL-4 levels. Postischemia, more CD8 T cells trafficked into postischemic kidneys of germ-free mice compared with control mice. Renal structural injury and functional decline following IRI were more severe in germ-free mice compared with control mice. When germ-free mice were conventionalized with the addition of bacteria to their diet, the extent of renal injury after IRI became equivalent to age-matched control mice, with similar numbers and phenotypes of T cells and NKT cells, as well as cytokine expression in both normal kidneys and postischemic kidneys of conventionalized germ-free mice and age-matched control mice. Thus microbial stimuli influence the phenotype of renal lymphocytes and the expression of cytokines of normal kidneys and also modulate the outcome of IRI. PMID- 19675180 TI - Endothelial-specific CYP4A2 overexpression leads to renal injury and hypertension via increased production of 20-HETE. AB - We have previously reported that adenoviral-mediated delivery of cytochrome P-450 (CYP) 4A2, which catalyzes the synthesis of 20-hydroxyeicosatetraenoic acid (20 HETE), results in endothelial dysfunction and hypertension in Sprague-Dawley (SD) rats (Wang JS, Singh H, Zhang F, Ishizuka T, Deng H, Kemp R, Wolin MS, Hintze TH, Abraham NG, Nasjletti A, Laniado-Schwartzman M. Circ Res 98: 962-969, 2006). In this study, we targeted the vascular endothelium by using a lentivirus construct expressing CYP4A2 under the control of the endothelium-specific promoter VE cadherin (VECAD-4A2) and examined the effect of long-term CYP4A2 overexpression on blood pressure and kidney function in SD rats. A bolus injection of VECAD-4A2 increased blood pressure (P < 0.001) by 26, 36, and 30 mmHg 10, 20, and 30 days postinjection, respectively. Arteries from VECAD-4A2-transduced rats produced increased levels of 20-HETE (P < 0.01), expressed lower levels of endothelial nitric oxide synthase (eNOS) and phosphorylated eNOS (p-eNOS) (P < 0.05), generated higher levels of superoxide anion, and displayed decreased relaxing responsiveness to acetylcholine (P < 0.05). Proteinuria increased by twofold in VECAD-4A2-transduced rats compared with controls. Treatment of VECAD-4A2 transduced rats with HET0016, an inhibitor of 20-HETE biosynthesis, not only attenuated the increase in blood pressure (P < 0.05) but also improved vascular function (acetylcholine-induced relaxations) and reduced plasma creatinine and proteinuria. HET0016 treatment decreased oxidative stress and increased the phosphorylated state of key proteins that regulate endothelial function, including eNOS, AKT, and AMPK. Collectively, these findings demonstrate that augmentation of vascular endothelial 20-HETE levels results in hypertension, endothelial dysfunction, and renal injury, which is offset by HET0016 through a reduction in vascular 20-HETE coupled with a lessening of oxidative stress and the amplification of pAKT, pAMPK, and p-eNOS levels leading to normalization of endothelial responses. PMID- 19675181 TI - Unlike each drug alone, lisinopril if combined with avosentan promotes regression of renal lesions in experimental diabetes. AB - In the present study, we evaluated the effect of simultaneously blocking angiotensin II synthesis and endothelin (ET)-1 activity as a multimodal intervention to implement renoprotection in overt diabetic nephropathy. Mechanisms underlying combined therapy effectiveness were addressed by investigating podocyte structure and function and glomerular barrier size selective properties. Uninephrectomized rats made diabetic by streptozotocin received orally placebo, lisinopril (12.5 mg/l), the ET(A) receptor antagonist avosentan (30 mg/kg), or their combination from 4 (when animals had proteinuria) to 8 mo. Proteinuria, renal damage, podocyte number, nephrin expression, and glomerular size selectivity by graded-size Ficoll molecule fractional clearance were assessed. Combined therapy normalized proteinuria, provided complete protection from tubulointerstitial damage, and induced regression of glomerular lesions, while only a partial renoprotection was achieved by each drug alone. Lisinopril plus avosentan restored to normal values the number of podocytes. Single therapies only limited podocyte depletion. Defective nephrin expression of diabetes was prevented by each drug. Altered glomerular size selectivity to large macromolecules of diabetic rats was remarkably improved by lisinopril and the combined treatment. Avosentan ameliorated peritubular capillary architecture and reduced interstitial inflammation and fibrosis. The ACE inhibitor and ET(A) receptor antagonist induced regression of glomerular lesions in overt diabetic nephropathy. Regression of renal disease was conceivably the result of the synergistic effect of the ACE inhibitor of preserving glomerular permselective properties and the ET(A) antagonist in improving tubulointerstitial changes. These findings provide mechanistic insights to explain the antiproteinuric effect of this combined therapy in diabetes. PMID- 19675182 TI - Adaptation of the length-active tension relationship in rabbit detrusor. AB - Studies have shown that the length-tension (L-T) relationships in airway and vascular smooth muscles are dynamic and can adapt to length changes over a period of time. Our prior studies have shown that the passive L-T relationship in rabbit detrusor smooth muscle (DSM) is also dynamic and that DSM exhibits adjustable passive stiffness (APS) characterized by a passive L-T curve that can shift along the length axis as a function of strain history and activation history. The present study demonstrates that the active L-T curve for DSM is also dynamic and that the peak active tension produced at a particular muscle length is a function of both strain and activation history. More specifically, this study reveals that the active L-T relationship, or curve, does not have a unique peak tension value with a single ascending and descending limb, but instead reveals that multiple ascending and descending limbs can be exhibited in the same DSM strip. This study also demonstrates that for DSM strips not stretched far enough to reveal a descending limb, the peak active tension produced by a maximal KCl-induced contraction at a short, passively slack muscle length of 3 mm was reduced by 58.6 +/- 4.1% (n = 15) following stretches to and contractions at threefold the original muscle length, 9 mm. Moreover, five subsequent contractions at the short muscle length displayed increasingly greater tension; active tension produced by the sixth contraction was 91.5 +/- 9.1% of that produced by the prestretch contraction at that length. Together, these findings indicate for the first time that DSM exhibits length adaptation, similar to vascular and airway smooth muscles. In addition, our findings demonstrate that preconditioning, APS and adaptation of the active L-T curve can each impact the maximum total tension observed at a particular DSM length. PMID- 19675184 TI - Penile-response profiles of sexual aggressors during phallometric testing. AB - This study analyzes sexual-preference profiles in a sample of 420 sexual aggressors who exhibited a valid profile during their initial phallometric assessment. The sexual stimuli used in the process were audiotapes describing sexual-offending scenarios. Two types of sexual stimuli sets were used: one developed for sexual aggressors against women and a second developed for sexual aggressors against children. Penile responses were recorded during stimulus presentation using a mercury-in-rubber strain gauge. Classification analyses (hierarchical and K-means clustering combination) were conducted separately for three groups of sexual aggressors: (a) sexual aggressors against children (n = 253), (b) sexual aggressors against women (n = 138), and (c) mixed sexual aggressors (n = 29). The sexual aggressors against children exhibited four penile response profiles, the sexual aggressors against women two penile-response profiles, and the mixed sexual aggressors only one penile-response profile. In addition, analyses carried out on randomly split subsamples established that the generated profiles for sexual aggressors against children and sexual aggressors against women were stable. PMID- 19675185 TI - Degradation and aggresome formation of the Gn tail of the apathogenic Tula hantavirus. AB - The cytoplasmic tails of envelope glycoprotein Gn of pathogenic hantaviruses but not of the apathogenic Prospect Hill virus (PHV) were recently reported to be proteasomally degraded in simian COS7 cells. Here, we show that the cytoplasmic tails of the glycoproteins of the apathogenic hantaviruses Tula virus (TULV) and PHV are also degraded through the ubiquitin-proteasome pathway, both in human HEK 293 and in simian Vero E6 cells. TULV Gn tails formed aggresomes in cells with proteasomal inhibitors. We conclude that degradation upon aggregation of Gn tails, which may represent a general cellular response to misfolded protein used by hantaviruses to control maturation of virions, is unrelated to pathogenicity. PMID- 19675183 TI - Regulation of rat intestinal Na-dependent phosphate transporters by dietary phosphate. AB - Hyperphosphatemia associated with chronic kidney disease is one of the factors that can promote vascular calcification, and intestinal P(i) absorption is one of the pharmacological targets that prevents it. The type II Na-P(i) cotransporter NaPi-2b is the major transporter that mediates P(i) reabsorption in the intestine. The potential role and regulation of other Na-P(i) transporters remain unknown. We have identified expression of the type III Na-P(i) cotransporter PiT 1 in the apical membrane of enterocytes. Na-P(i) transport activity and NaPi-2b and PiT-1 proteins are mostly expressed in the duodenum and jejunum of rat small intestine; their expression is negligible in the ileum. In response to a chronic low-P(i) diet, there is an adaptive response restricted to the jejunum, with increased brush border membrane (BBM) Na-P(i) transport activity and NaPi-2b, but not PiT-1, protein and mRNA abundance. However, in rats acutely switched from a low- to a high-P(i) diet, there is an increase in BBM Na-P(i) transport activity in the duodenum that is associated with an increase in BBM NaPi-2b protein abundance. Acute adaptive upregulation is restricted to the duodenum and induces an increase in serum P(i) that produces a transient postprandial hyperphosphatemia. Our study, therefore, indicates that Na-P(i) transport activity and NaPi-2b protein expression are differentially regulated in the duodenum vs. the jejunum and that postprandial upregulation of NaPi-2b could be a potential target for treatment of hyperphosphatemia. PMID- 19675186 TI - Domain organization of the N-terminal portion of hordeivirus movement protein TGBp1. AB - Three 'triple gene block' proteins known as TGBp1, TGBp2 and TGBp3 are required for cell-to-cell movement of plant viruses belonging to a number of genera including Hordeivirus. Hordeiviral TGBp1 interacts with viral genomic RNAs to form ribonucleoprotein (RNP) complexes competent for translocation between cells through plasmodesmata and over long distances via the phloem. Binding of hordeivirus TGBp1 to RNA involves two protein regions, the C-terminal NTPase/helicase domain and the N-terminal extension region. This study demonstrated that the extension region of hordeivirus TGBp1 consists of two structurally and functionally distinct domains called the N-terminal domain (NTD) and the internal domain (ID). In agreement with secondary structure predictions, analysis of circular dichroism spectra of the isolated NTD and ID demonstrated that the NTD represents a natively unfolded protein domain, whereas the ID has a pronounced secondary structure. Both the NTD and ID were able to bind ssRNA non specifically. However, whilst the NTD interacted with ssRNA non-cooperatively, the ID bound ssRNA in a cooperative manner. Additionally, both domains bound dsRNA. The NTD and ID formed low-molecular-mass oligomers, whereas the ID also gave rise to high-molecular-mass complexes. The isolated ID was able to interact with both the NTD and the C-terminal NTPase/helicase domain in solution. These data demonstrate that the hordeivirus TGBp1 has three RNA-binding domains and that interaction between these structural units can provide a basis for remodelling of viral RNP complexes at different steps of cell-to-cell and long distance transport of virus infection. PMID- 19675187 TI - Bovine papillomavirus type 1 oncoprotein E5 inhibits equine MHC class I and interacts with equine MHC I heavy chain. AB - Bovine papillomavirus type 1 is one of the aetiological agents of equine sarcoids. The viral major oncoprotein E5 is expressed in virtually all sarcoids, sarcoid cell lines and in vitro-transformed equine fibroblasts. To ascertain whether E5 behaves in equine cells as it does in bovine cells, we introduced the E5 open reading frame into fetal equine fibroblasts (EqPalF). As observed in primary bovine fibroblasts (BoPalF), E5 by itself could not immortalize EqPalF and an immortalizing gene, such as human telomerase (hTERT/hT), was required for the cells to survive selection. The EqPalF-hT-1E5 cells were morphologically transformed, elongated with many pseudopodia and capable of forming foci. Equine major histocompatibility complex class I (MHC I) was inhibited in these cells at least at two levels: transcription of MHC I heavy chain was inhibited and the MHC I complex was retained in the Golgi apparatus and prevented from reaching the cell surface. We conclude that, as in bovine cells and tumours, E5 is a player in the transformation of equine cells and the induction of sarcoids, and a potential major cause of MHC I downregulation and hence poor immune clearance of tumour cells. PMID- 19675188 TI - Establishment of an insect cell clone that harbours a partial baculoviral genome and is resistant to homologous virus infection. AB - After serially undiluted passage of Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV), persistently infected Se301 cells were established. A cell strain, in which no polyhedra or viral particles were observed, was cloned and designated P8-Se301-C1. The P8-Se301-C1 cells are morphologically similar to but grow slower than Se301 cells and they can homologously interfere with SeMNPV. PCR analysis showed that SeMNPV ie-0 and polyhedrin genes were present but DNA polymerase and orf67 genes were absent in P8-Se301-C1, suggesting that the cells harbour incomplete SeMNPV genomes. Dot blot analysis demonstrated that 0.32+/-0.16 ng SeMNPV DNA was present in 1.25 x 10(5) P8-Se301-C1 cells. A quantitative real-time PCR assay showed that there were 13.2+/-4.3 copies of the SeMNPV polyhedrin gene in each cell. Nested RT-PCR demonstrated the presence of SeMNPV polyhedrin transcripts in P8-Se301-C1 cells. The fact that P8-Se301-C1 cells carry low levels of partial viral genome but do not produce viral progeny suggests a latent-like viral infection in the cells. PMID- 19675189 TI - Prevalence and genetic variability of tick-borne encephalitis virus in host seeking Ixodes ricinus in northern Italy. AB - Tick-borne encephalitis (TBE) is a severe disease that has been endemic in north east Italy since 1992. Over the past two decades, there has been an increase in the number of human cases reported in many European countries, including Italy. To assess the current TBE infection risk, questing ticks were collected from known TBE foci, as well as from a site in northern Italy where no human infections have been reported previously. A total of 1739 Ixodes ricinus (1485 nymphs and 254 adults) was collected and analysed for TBEV prevalence by a real time RT-PCR targeting the 3' untranslated region. Phylogenetic analyses of the partial envelope gene were conducted on two newly sequenced TBE virus (TBEV) strains and 28 previously published sequences to investigate the genealogical relationships of the circulating TBEV strains. These phylogenetic analyses confirmed a previous report that the European TBEV subtype is the only subtype circulating within the TBE foci in north-east Italy. Interestingly, nucleotide sequence analysis revealed a high degree of divergence (mean 2.54 %) between the TBEV strains recovered in the Italian province of Trento, despite the circulation of a single TBEV subtype. This elevated genetic variability within a single TBE focus may reflect local differences in the long-standing evolutionary dynamics of TBEV at this site relative to previously characterized sites, or more recent and continuous reintroduction of various TBEV strains. PMID- 19675190 TI - Mutational analysis of eggplant latent viroid RNA processing in Chlamydomonas reinhardtii chloroplast. AB - Viroids of the family Avsunviroidae, such as eggplant latent viroid (ELVd), contain hammerhead ribozymes and replicate in the chloroplasts of the host plant through an RNA-based symmetrical rolling-circle mechanism in which oligomeric RNAs of both polarity are processed to monomeric linear RNAs (by cleavage) and to monomeric circular RNAs (by ligation). Using an experimental system consisting of transplastomic lines of the alga Chlamydomonas reinhardtii, a mutational analysis of sequence and structural elements in the ELVd molecule that are involved in transcript processing in vivo in a chloroplastic context was carried out. A collection of six insertion and three deletion ELVd mutants was created and expressed in C. reinhardtii chloroplast. All mutants cleaved efficiently except for the control with an insertion inside the hammerhead ribozyme domain, supporting the prediction that this domain is necessary and sufficient to mediate transcript cleavage in vivo. However, two deletion mutants that cleaved efficiently showed ligation defects, indicating that during RNA circularization, other parts of the molecule are involved in addition to the hammerhead ribozyme domain. This is probably a quasi double-stranded structure present in the central part of the molecule which contains the ligation site in an internal loop. However, the mutations prevented the viroid from infecting its natural host, eggplant, indicating that they affected other essential functions in ELVd infectious cycle. The insertion in the terminal loop of the right upper hairpin of ELVd did not have this effect; it was tolerated and partially maintained in the progeny. PMID- 19675191 TI - The 2b protein of cucumber mosaic virus is essential for viral infection of the shoot apical meristem and for efficient invasion of leaf primordia in infected tobacco plants. AB - It has been reported previously that a 2b protein-defective mutant of the cucumber mosaic virus (CMV) Pepo strain (Delta 2b) induces only mild symptoms in systemically infected tobacco plants. To clarify further the role of the 2b protein as an RNA silencing suppressor in mosaic symptom expression during CMV infection, this study monitored the sequential distribution of Delta 2b in the shoot meristem and leaf primordia (LP) of inoculated tobacco. Time-course histochemical observations revealed that Delta 2b was distributed in the shoot meristem at 7 days post-inoculation (p.i.), but could not invade shoot apical meristem (SAM) and quickly disappeared from the shoot meristem, whereas wild-type (Pepo) transiently appeared in SAM from 4 to 10 days p.i. In LP, Delta 2b signals were detected only at 14 and 21 days p.i., whereas dense Pepo signals were observed in LP from 4 to 18 days p.i. Northern blot analysis showed that small interfering RNA (siRNA) derived from Delta 2b RNA accumulated earlier in the shoot meristem and LP than that of Pepo. However, a similar amount of siRNA was detected in both Pepo- and Delta 2b-infected plants at late time points. Tissue printing analysis of the inoculated leaves indicated that the areas infected by Pepo increased faster than those infected by Delta 2b, whereas accumulation of Delta 2b in protoplasts was similar to that of Pepo. These findings suggest that the 2b protein of the CMV Pepo strain determines virulence by facilitating the distribution of CMV in the shoot meristem and LP via prevention of RNA silencing and/or acceleration of cell-to-cell movement. PMID- 19675192 TI - Association between neighborhood-level deprivation and disability in a community sample of people with diabetes. AB - OBJECTIVE: The objective of the present study was to analyze the association between neighborhood deprivation and self-reported disability in a community sample of people with type 2 diabetes. RESEARCH DESIGN AND METHODS: Random digit dialing was used to select a sample of adults with self-reported diabetes aged 18 80 years in Quebec, Canada. Health status was assessed by the World Health Organization Disability Assessment Schedule II. Material and social deprivation was measured using the Pampalon index, which is based on the Canadian Census. Potential risk factors for disability included sociodemographic characteristics, socioeconomic status, social support, lifestyle-related factors (smoking, physical activity, and BMI), health care-related problems, duration of diabetes, insulin use, and diabetes-specific complications. RESULTS: There was a strong association between disability and material and social deprivation in our sample (n = 1,439): participants living in advantaged neighborhoods had lower levels of disability than participants living in disadvantaged neighborhoods. The means +/- SD disability scores for men were 7.8 +/- 11.8, 12.0 +/- 11.8, and 18.1 +/- 19.4 for low, medium, and high deprivation areas, respectively (P < 0.001). The disability scores for women were 13.4 +/- 12.4, 14.8 +/- 15.9, and 18.9 +/- 16.2 for low, medium, and high deprivation areas, respectively (P < 0.01). Neighborhood deprivation was associated with disability even after controlling for education, household income, sociodemographic characteristics, race, lifestyle-related behaviors, social support, diabetes-related variables, and health care access problems. CONCLUSIONS: The inclusion of neighborhood characteristics might be an important step in the identification and interpretation of risk factors for disability in diabetes. PMID- 19675193 TI - Sustained benefit of continuous glucose monitoring on A1C, glucose profiles, and hypoglycemia in adults with type 1 diabetes. AB - OBJECTIVE: To evaluate long-term effects of continuous glucose monitoring (CGM) in intensively treated adults with type 1 diabetes. RESEARCH DESIGN AND METHODS: We studied 83 of 86 individuals >or=25 years of age with type 1 diabetes who used CGM as part of a 6-month randomized clinical trial in a subsequent 6-month extension study. RESULTS After 12 months, median CGM use was 6.8 days per week. Mean change in A1C level from baseline to 12 months was -0.4 +/- 0.6% (P < 0.001) in subjects with baseline A1C >or=7.0%. A1C remained stable at 6.4% in those with baseline A1C <7.0%. The incidence rate of severe hypoglycemia was 21.8 and 7.1 events per 100 person-years in the first and last 6 months, respectively. Time per day with glucose levels in the range of 71-180 mg/dl increased significantly (P = 0.02) from baseline to 12 months. CONCLUSIONS: In intensively treated adults with type 1 diabetes, CGM use and benefit can be sustained for 12 months. PMID- 19675194 TI - Vitamin D levels in subjects with and without type 1 diabetes residing in a solar rich environment. AB - OBJECTIVE: Previous studies, largely in northern Europe, have suggested an association between type 1 diabetes and reduced serum 25-hydroxy(OH) vitamin D levels, a concept we tested in individuals residing in a solar-rich region (Florida). RESEARCH DESIGN AND METHODS: Serum samples from 415 individuals residing in Florida were cross-sectionally analyzed: 153 control subjects, 46 new onset type 1 diabetic patients, 110 established type 1 diabetic patients (samples >or=5 months from diagnosis), and 106 first-degree relatives of the diabetic patients. RESULTS: In this study, 25-OH vitamin D levels (median, range, interquartile range [IQR]) were similar among control subjects (20.1, below detection [bd]-163.5, 13.0-37.4 ng/ml), new-onset type 1 diabetic patients (21.2, bd-48.6, 12.2-30.2 ng/ml), established type 1 diabetic patients (23.2, bd-263.8, 13.8-33.9 ng/ml), and first-degree relatives (22.2, bd-59.9, 12.7-33.1 ng/ml) (P = 0.87). Mean 25-OH vitamin D levels were less than the optimal World Health Organization level of 30 ng/ml in all study groups. CONCLUSIONS: Reduced serum 25 OH vitamin D levels were not specifically associated with type 1 diabetes. The uniform suboptimal 225-OH vitamin D levels, despite residence in a zone with abundant sunshine, support additional dietary vitamin D fortification practices. PMID- 19675195 TI - Obstetric and perinatal outcomes in type 1 diabetic pregnancies: A large, population-based study. AB - OBJECTIVE: To perform comparative analyses of obstetric and perinatal outcomes between type 1 diabetic pregnancies and the general obstetric population in Sweden between 1991 and 2003. RESEARCH DESIGN AND METHODS: This was a population based study. Data were obtained from the Medical Birth Registry, covering >98% of all pregnancies in Sweden. A total of 5,089 type 1 diabetic pregnancies and 1,260,207 control pregnancies were included. Odds ratios (ORs) were adjusted for group differences in maternal age, parity, BMI, chronic hypertensive disease, smoking habits, and ethnicity. RESULTS: In type 1 diabetes, preeclampsia was significantly more frequent (OR 4.47 [3.77-5.31]) as was delivery by cesarean section (5.31 [4.97-5.69]) compared with results for the general population. Stillbirth (3.34 [2.46-4.55]), perinatal mortality (3.29 [2.50-4.33]), and major malformations (2.50 [2.13-2.94]) were more common in type 1 diabetic than in control pregnancies. The risk of very preterm birth (<32 gestational weeks) was also higher among type 1 diabetic women (3.08 [2.45-3.87]). The incidence of fetal macrosomia (birth weight >or=2 SD above the mean) was increased in the diabetic group (11.45 [10.61-12.36]). CONCLUSIONS: Type 1 diabetes in pregnancy is still associated with considerably increased rates of adverse obstetric and perinatal outcomes. The eightfold increased risk for fetal macrosomia in type 1 diabetic pregnancies is unexpected and warrants further investigation. PMID- 19675197 TI - Diabetic retinopathy is associated with elevated serum asymmetric and symmetric dimethylarginines. AB - OBJECTIVE: Asymmetric dimethylarginine (ADMA), symmetric dimethylarginine (SDMA), and l-arginine directly influence nitric oxide production. Our objective was to test whether serum ADMA, SDMA, or l-arginine levels correlate with diabetic retinopathy subtype or severity. RESEARCH DESIGN AND METHODS: A total of 162 subjects with type 1 diabetes and 343 with type 2 diabetes, of whom 329 subjects had no diabetic retinopathy, 27 had nonproliferative diabetic retinopathy (NPDR), 101 had proliferative diabetic retinopathy (PDR), and 107 had clinically significant macular edema (CSME), were recruited. Blinding diabetic retinopathy was defined as severe NPDR, PDR, or CSME. Serum ADMA, SDMA, and l-arginine concentrations were determined by mass spectroscopy. RESULTS: In multivariate analysis, blinding diabetic retinopathy, PDR, and nephropathy were associated with significantly increased serum levels of ADMA (P < 0.001), SDMA (P < 0.001), and l-arginine (P = 0.001). Elevated ADMA (P < 0.001) and SDMA (P < 0.001) were also significantly associated with CSME. CONCLUSIONS: Severe forms of diabetic retinopathy are associated with elevated serum ADMA, SDMA, and l-arginine. Further investigation is required to determine whether these findings are of clinical relevance. PMID- 19675196 TI - Determinants of quality in diabetes care process: The population-based Torino Study. AB - OBJECTIVE: To investigate the role of clinical and socioeconomic variables as determinants of adherence to recommended diabetes care guidelines and assess differences in the process of care between diabetologists and general practitioners. RESEARCH DESIGN AND METHODS: We identified diabetic residents in Torino, Italy, as of 31 July 2003, using multiple independent data sources. We collected data on several laboratory tests and specialist medical examinations registered during the subsequent 12 months and performed regression analyses to identify associations with quality-of-care indicators based on existing guidelines. RESULTS: After 1 year, only 35.8% of patients had undergone a comprehensive assessment. In the multivariate models, factors independently and significantly associated with lower quality of care were age >or=75 years (prevalence rate ratio [PRR] 0.66 [95% CI 0.61-0.70]) and established cardiovascular disease (0.89 [0.86-0.93]). Disease severity (PRR for insulin treated patients 1.45 [1.38-1.53]) and diabetologist consultation (PRR 3.34 [3.17 3.53]) were positively associated with high quality of care. No clear association emerged between sex and socioeconomic status. These differences were strongly reduced in patients receiving diabetologist care compared with patients receiving general practitioner care only. CONCLUSIONS: Despite widespread availability of guidelines and simple screening procedures, a nonnegligible portion of the diabetic population, namely elderly individuals and patients with less severe forms of the disease, are not properly cared for. As practitioners in diabetes centers are more likely to adhere to guidelines than general practitioners, quality in the diabetes care process can be improved by increasing the intensity of disease management programs, with greater participation by general practitioners. PMID- 19675198 TI - Correlation between albuminuria and spontaneous platelet microaggregate formation in type 2 diabetic patients. AB - OBJECTIVE: Albuminuria in type 2 diabetic patients is a risk factor for cardiovascular disease. We investigated the correlation between albuminuria and spontaneous microaggregation of platelets (SMAP) formed under shear stress. RESEARCH DESIGN AND METHODS: The study subjects were 401 type 2 diabetic individuals (252 with normoalbuminuria and 149 with albuminuria) who were examined for SMAP under conditions of shear stress only (no agonist stimulation) and the reversibility of platelet microaggregation after stimulation with 1 mumol/l ADP, measured by a laser light-cattering method. Active glycoprotein IIb/IIIa (GPIIb/IIIa) and P-selectin expression levels on platelets as an index of platelet activation were measured by whole-blood flow cytometry. RESULTS: SMAP formation was noted in 53% of diabetic patients. All patients with SMAP showed an irreversible pattern of platelet microaggregates by a low dose of ADP. SMAP was observed in 75% of diabetic subjects with albuminuria and in 39% of those with normoalbuminuria. Multivariate logistic regression analysis identified urinary albumin excretion rate and brachial-ankle pulse-wave velocity as independent factors associated with SMAP. The degree of SMAP correlated with active GPIIb/IIIa (gamma = 0.59, P < 0.001) and P-selectin (gamma = 0.55, P < 0.001) expression levels. These early-activated platelet profiles were significantly inhibited in albuminuric patients with aspirin intake, although the effect was incomplete. CONCLUSIONS: Our study demonstrated an independent association between albuminuria and early changes in activated platelet profiles of type 2 diabetic patients. Further follow-up and intervention studies are needed to establish whether the inhibition of SMAP affects the course of cardiovascular disease in type 2 diabetic patients. PMID- 19675200 TI - Eating fish and risk of type 2 diabetes: A population-based, prospective follow up study. AB - OBJECTIVE: To investigate the relation between total fish, type of fish (lean and fatty), and eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) intake and risk of type 2 diabetes in a population-based cohort. RESEARCH DESIGN AND METHODS: The analysis included 4,472 Dutch participants aged >or=55 years without diabetes at baseline. Dietary intake was assessed with a semiquantitative food frequency questionnaire. Hazard ratios (relative risk [RR]) with 95% CIs were used to examine risk associations adjusted for age, sex, lifestyle, and nutritional factors. RESULTS: After 15 years of follow-up, 463 participants developed type 2 diabetes. Median fish intake, mainly lean fish (81%), was 10 g/day. Total fish intake was associated positively with risk of type 2 diabetes; the RR was 1.32 (95% CI 1.02-1.70) in the highest total fish group (>or=28 g/day) compared with that for non-fish eaters (P(trend) = 0.04). Correspondingly, lean fish intake tended to be associated positively with type 2 diabetes (RR highest group [>or=23 g/day] 1.30 [95% CI 1.01-1.68]; P(trend) = 0.06), but fatty fish was not. No association was observed between EPA and DHA intake and type 2 diabetes (RR highest group [>or=149.4 mg/day] 1.22 [0.97-1.53]). With additional adjustment for intake of selenium, cholesterol, and vitamin D, this RR decreased to 1.05 (0.80-1.38; P(trend) = 0.77). CONCLUSIONS: The findings do not support a beneficial effect of total fish, type of fish, or EPA and DHA intake on the risk of type 2 diabetes. Alternatively, other dietary components, such as selenium, and unmeasured contaminants present in fish might explain our results. PMID- 19675199 TI - Association of type 1 diabetes with month of birth among U.S. youth: The SEARCH for Diabetes in Youth Study. AB - OBJECTIVE: Seasonal environment at birth may influence diabetes incidence in later life. We sought evidence for this effect in a large sample of diabetic youth residing in the U.S. RESEARCH DESIGN AND METHODS: We compared the distribution of birth months within the SEARCH for Diabetes in Youth Study (SEARCH study) with the monthly distributions in U.S. births tabulated by race for years 1982-2005. SEARCH study participants (9,737 youth with type 1 diabetes and 1,749 with type 2 diabetes) were identified by six collaborating U.S. centers. RESULTS: Among type 1 diabetic youth, the percentage of observed to expected births differed across the months (P = 0.0092; decreased in October February and increased in March-July). Their smoothed birth-month estimates demonstrated a deficit in November-February births and an excess in April-July births (smoothed May versus January relative risk [RR] = 1.06 [95% CI 1.02 1.11]). Stratifications by sex or by three racial groups showed similar patterns relating type 1 diabetes to month of birth. Stratification by geographic regions showed a peak-to-nadir RR of 1.10 [1.04-1.16] in study regions from the northern latitudes (Colorado, western Washington State, and southern Ohio) but no birth month effect (P > 0.9) in study regions from more southern locations. Among type 2 diabetic youth, associations with birth month were inconclusive. CONCLUSIONS: Spring births were associated with increased likelihood of type 1 diabetes but possibly not in all U.S. regions. Causal mechanisms may involve factors dependent on geographic latitude such as solar irradiance, but it is unknown whether they influence prenatal or early postnatal development. PMID- 19675201 TI - Prevalence and management of diabetes in Korean adults: Korea National Health and Nutrition Examination Surveys 1998-2005. AB - OBJECTIVE: This research investigated recent changes in the prevalence and management status of diabetes among Korean adults. RESEARCH DESIGN AND METHODS: The Korea National Health and Nutrition Examination Survey (KNHANES), a nationwide survey examining the general health and nutrition status of the Korean people, was conducted in 1998, 2001, and 2005. Using the first (1998; n = 5,645), second (2001; n = 4,154), and third (2005; n = 4,628) KNHANES datasets, in the present study, we estimated the prevalence of diabetes among Korean adults (aged >or=30 years), the proportions of known cases of diabetes, and the proportions of well-controlled cases of diabetes, as defined by either the American Diabetes Association (A1C <7%) or the International Diabetes Federation guidelines (A1C <6.5%). RESULTS: In 2005, the prevalence of diabetes was estimated to be 9.1% (approximately 2.58 million people: 10.2% of men and 7.9% of women), including 6.2% with known diabetes and 2.9% with newly diagnosed diabetes. The prevalence of impaired fasting glucose was 17.4% (approximately 4.94 million people). The proportion of known cases of diabetes drastically increased from 23.2% in 1998 to 41.2% in 2001 and 68.0% in 2005 (P < 0.0001). Among known diabetic patients in 2005, 43.5 and 22.9% had A1C levels <7.0 and <6.5%, respectively. CONCLUSIONS: The overall prevalence of diabetes in Korea has not changed significantly between 1998 and 2005. Physician diagnosis and treatment rates of diabetes have significantly improved during this period, but glycemic control was still poorer than that in other developed countries. PMID- 19675202 TI - Dietary patterns and glucose tolerance abnormalities in Chinese adults. AB - OBJECTIVE: To investigate the association of the dietary pattern with the presence of newly diagnosed glucose tolerance abnormalities among Chinese adults. RESEARCH DESIGN AND METHODS: A total of 20,210 adults aged 45-69 years from the 2002 China National Nutrition and Health Survey were included. Information on dietary intake was collected using a validated food frequency questionnaire. Factor analysis and cluster analysis were used to identify the food factors and dietary pattern clusters. RESULTS: Four dietary pattern clusters were identified ("Green Water," "Yellow Earth," "Western Adopter," and "New Affluence"). The prevalence of glucose tolerance abnormalities ranged from 3.9% in the Green Water to 8.0% in the New Affluence. After adjustment for area, age, sex, current smoking, and physical activity, subjects in the Yellow Earth cluster (prevalence ratio 1.22 [95% CI 1.04-1.43]) and New Affluence cluster (2.05 [1.76-2.37]) had significantly higher prevalence rates compared with those for the Green Water cluster. After further adjustment for BMI and waist-to-height ratio, the elevated risk in the New Affluence remained statistically significant. CONCLUSIONS: Dietary patterns and food factors are associated with the presence of glucose tolerance abnormalities in China, even independent of obesity. A New Affluence diet is an important modifiable risk factor, which needs attention from the prevention point of view. PMID- 19675203 TI - In vivo insulin sensitivity and lipoprotein particle size and concentration in black and white children. AB - OBJECTIVE: To examine sex-specific black/white differences in lipoprotein profile and the role of visceral adiposity and to assess the relationship between insulin sensitivity and lipoprotein profiles in each group. RESEARCH DESIGN AND METHODS: Fasting lipoprotein particle size and concentration and visceral adipose tissue (VAT) were determined in 226 children (117 black, 101 male) aged 8 to <18 years. The relationship between lipoproteins and insulin sensitivity was evaluated in a subset of 194 children (100 black, 88 male) who underwent a hyperinsulinemic euglycemic clamp. RESULTS: Black male children had smaller VLDL and black female children had larger HDL size than their white counterparts. Overall, blacks had larger LDL size with no sex-specific race differences. After adjusting for VAT and sex, only VLDL size and concentrations remained significantly favorable in blacks. Analysis of lipoprotein particle size and concentration across insulin sensitivity quartiles revealed that in both racial groups, the most insulin resistant children had higher concentrations of small dense LDL, small HDL, and large VLDL and smaller LDL and HDL sizes than their more insulin-sensitive counterparts. CONCLUSIONS: The previously reported favorable lipoprotein profiles in black versus white children is partly due to race differences in VAT. In both groups, however, the most insulin-resistant youths have a high-risk atherogenic profile of small dense LDL, small HDL, and large VLDL, akin to the atherogenic lipoprotein pattern in adults with coronary artery disease. PMID- 19675204 TI - Insulin sensitivity as an independent predictor of fat mass gain in Hispanic adolescents. AB - OBJECTIVE: The purpose of this study was to examine the relationship between changes in insulin sensitivity and subsequent changes in fat mass in obese Hispanic children over 3 consecutive years. RESEARCH DESIGN AND METHODS: In a longitudinal research design, insulin sensitivity (S(i)) of 96 research participants was determined at baseline and 1 year later. Body adiposity was assessed at four assessments. RESULTS: The change in S(i) during the first year of the study was a significant predictor of further fat mass development (P < 0.05). Considering different directions of S(i) change, S(i) was a strong predictor for further fat mass development only in the group that decreased their S(i) (P < 0.05). CONCLUSIONS: The results show that the direction of change in insulin sensitivity at an early age is an important independent predictor for further fat mass development and emphasize the importance of insulin sensitivity as a primary target for long-term obesity prevention, as well as the significance of early age intervention. PMID- 19675205 TI - Blunted counterregulatory hormone responses to hypoglycemia in young children and adolescents with well-controlled type 1 diabetes. AB - OBJECTIVE: Hypoglycemia in young children with type 1 diabetes is an acute complication of intensive insulin therapy and is commonly observed in the absence of signs or symptoms. The effect of intensive treatment and patient age on sympathoadrenal responses has not been established in youth with type 1 diabetes because of difficulties in testing procedures. RESEARCH DESIGN AND METHODS: We developed a standardized inpatient continuous subcutaneous insulin infusion protocol to produce a progressive fall in plasma glucose concentrations in insulin pump-treated patients. Plasma glucose and counterregulatory hormone concentrations were measured in 14 young children (3 to <8 years, A1C 7.7 +/- 0.6%) vs. 14 adolescents (12 to <18 years, A1C 7.6 +/- 0.8%). RESULTS: Plasma glucose decreased to similar nadir concentrations in the two groups. Four young children and four adolescents never had an epinephrine response. In the four young children and five adolescents who had a modest epinephrine response, this only occurred when plasma glucose fell to <60 mg/dl. In evaluating symptom scores, 29% of parents of young children felt that their child looked hypoglycemic, even at the lowest plasma glucose concentrations. Adolescents were better able to detect symptoms of hypoglycemia. In comparison with our data, epinephrine response to hypoglycemia in 14 nondiabetic adolescents studied at the Children's Hospital of Pittsburgh was higher. CONCLUSIONS: These data suggest that even young children and adolescents with type 1 diabetes are prone to develop hypoglycemia-associated autonomic failure regardless of duration. Whether these abnormalities can be reversed using continuous glucose monitoring and closed-loop insulin delivery systems awaits further study. PMID- 19675206 TI - Factors predictive of use and of benefit from continuous glucose monitoring in type 1 diabetes. AB - OBJECTIVE: To evaluate factors associated with successful use of continuous glucose monitoring (CGM) among participants with intensively treated type 1 diabetes in the Juvenile Diabetes Research Foundation Continuous Glucose Monitoring Randomized Clinical Trial. RESEARCH DESIGN AND METHODS: The 232 participants randomly assigned to the CGM group (165 with baseline A1C >or=7.0% and 67 with A1C <7.0%) were asked to use CGM on a daily basis. The associations of baseline factors and early CGM use with CGM use >or=6 days/week in the 6th month and with change in A1C from baseline to 6 months were evaluated in regression models. RESULTS: The only baseline factors found to be associated with greater CGM use in month 6 were age >or=25 years (P < 0.001) and more frequent self-reported prestudy blood glucose meter measurements per day (P < 0.001). CGM use and the percentage of CGM glucose values between 71 and 180 mg/dl during the 1st month were predictive of CGM use in month 6 (P < 0.001 and P = 0.002, respectively). More frequent CGM use was associated with a greater reduction in A1C from baseline to 6 months (P < 0.001), a finding present in all age-groups. CONCLUSIONS: After 6 months, near-daily CGM use is more frequent in intensively treated adults with type 1 diabetes than in children and adolescents, although in all age-groups near-daily CGM use is associated with a similar reduction in A1C. Frequency of blood glucose meter monitoring and initial CGM use may help predict the likelihood of long-term CGM benefit in intensively treated patients with type 1 diabetes of all ages. PMID- 19675207 TI - Cholesterol-dependent cytolysins induce rapid release of mature IL-1beta from murine macrophages in a NLRP3 inflammasome and cathepsin B-dependent manner. AB - CDC are exotoxins secreted by many Gram-positive bacteria that bind cholesterol and oligomerize to form pores in eukaryotic cell membranes. We demonstrate that CDC TLO induces caspase-1 cleavage and the rapid release of IL-1beta from LPS primed murine BMDM. IL-1beta secretion depends on functional toxin pore formation, as free cholesterol, which prevents TLO binding to cell membranes, blocks the cytokine release. Secretion of the mature forms of IL-1beta and caspase-1 occurs only at lower TLO doses, whereas at a higher concentration, cells release the biologically inactive proforms. IL-1beta release at a low TLO dose requires potassium efflux, calcium influx, and the activities of calcium independent PLA(2), caspase-1, and cathepsin B. Additionally, mature IL-1beta release induced by a low TLO dose is dependent on the NLRP3 inflammasome, and pro IL-1beta release induced by a high TLO dose occurs independently of NLRP3. These results further elucidate a mechanism of CDC-induced IL-1beta release and suggest a novel, immune evasion strategy in which IL-1beta-containing macrophages might release primarily inactive cytokine following exposure to high doses of these toxins. PMID- 19675209 TI - The Ldb1 and Ldb2 transcriptional cofactors interact with the Ste20-like kinase SLK and regulate cell migration. AB - Cell migration involves a multitude of signals that converge on cytoskeletal reorganization, essential for development, immune responses, and tissue repair. Here, we show that the microtubule-associated Ste20 kinase SLK, required for cell migration, interacts with the LIM domain binding transcriptional cofactor proteins Ldb1/CLIM2 and Ldb2/CLIM1/NLI. We demonstrate that Ldb1 and 2 bind directly to the SLK carboxy-terminal AT1-46 homology domain in vitro and in vivo. We find that Ldb1 and -2 colocalize with SLK in migrating cells and that both knockdown and overexpression of either factor results in increased motility. Supporting this, knockdown of Ldb1 increases focal adhesion turnover and enhances migration in fibroblasts. We propose that Ldb1/2 function to maintain SLK in an inactive state before its activation. These findings highlight a novel function for Ldb1 and -2 and expand their role to include the control of cell migration. PMID- 19675208 TI - Brewing complications: the effect of acute ethanol exposure on wound healing. AB - Ethanol consumption is linked to a higher incidence of traumatic wounds and increases the risk for morbidity and mortality following surgical or traumatic injury. One of the most profound effects of acute ethanol exposure on wound healing occurs during the inflammatory response, and altered cytokine production is a primary component. Acute ethanol exposure also impairs the proliferative response during healing, causing delays in epithelial coverage, collagen synthesis, and blood vessel regrowth. The accumulated data support the paradigm that acute ethanol intoxication prior to injury significantly diminishes a patient's ability to heal efficiently. PMID- 19675210 TI - Golgi-associated cPLA2alpha regulates endothelial cell-cell junction integrity by controlling the trafficking of transmembrane junction proteins. AB - In endothelial cells specifically, cPLA2alpha translocates from the cytoplasm to the Golgi complex in response to cell confluence. Considering the link between confluence and cell-cell junction formation, and the emerging role of cPLA2alpha in intracellular trafficking, we tested whether Golgi-associated cPLA2alpha is involved in the trafficking of junction proteins. Here, we show that the redistribution of cPLA2alpha from the cytoplasm to the Golgi correlates with adherens junction maturation and occurs before tight junction formation. Disruption of adherens junctions using a blocking anti-VE-cadherin antibody reverses the association of cPLA2alpha with the Golgi. Silencing of cPLA2alpha and inhibition of cPLA2alpha enzymatic activity using various inhibitors result in the diminished presence of the transmembrane junction proteins VE-cadherin, occludin, and claudin-5 at cell-cell contacts, and in their accumulation at the Golgi. Altogether, our data support the idea that VE-cadherin triggers the relocation of cPLA2alpha to the Golgi and that in turn, Golgi-associated cPLA2alpha regulates the transport of transmembrane junction proteins through or from the Golgi, thereby controlling the integrity of endothelial cell-cell junctions. PMID- 19675211 TI - Evaluation of PCR-based testing for surveillance of KPC-producing carbapenem resistant members of the Enterobacteriaceae family. AB - The spread of carbapenem-resistant members of the Enterobacteriaceae family (CRE) harboring carbapenemases is an emerging public health threat. As KPC-producing Klebsiella species are endemic in our tertiary care hospital, we aimed to evaluate a PCR-based surveillance test for identification of rectal carriage of KPC-producing CRE. We conducted a surveillance study between May and December 2007. Rectal swabs were collected from patients known to harbor CRE and from contacts of newly discovered patients harboring CRE. Specimens were evaluated by culture and by PCR analysis for blaKPC and were defined as positive if CRE was cultured and blaKPC was identified. Discrepant results between the culture and PCR analysis were resolved by subculturing, repeating the PCR, and performing a hydrolysis assay. Positive CRE cultures prior or subsequent to the time of sampling for the study were also taken into consideration. Sensitivity, specificity, and time to result were calculated. A total of 755 swabs were included. Concordant results were documented for 735 specimens; 51 were positive as determined by both PCR and culture. Discrepancies existed for 20 swabs; 9 were blaKPC negative and CRE culture positive, and 11 were blaKPC positive and CRE culture negative. After repeat testing, a total of 64 samples were classified as blaKPC-positive CRE. The sensitivity and specificity of the PCR analysis were 92.2% and 99.6%, respectively, and those of the culture were 87.5% and 99.4%, respectively. Over the last 3 months of the study, the sensitivity of the PCR improved to 96.3%, versus 77.8% for culture. Time to result was 30 h for the PCR and 60 h (negative) and 75 h (positive) for the CRE culture. blaKPC PCR-based testing is a useful method for the surveillance of KPC-producing CRE. Its main advantage over culturing is a shorter time to result, and it may prove to be more sensitive. PMID- 19675212 TI - Genomic analyses of recombinant adenovirus type 11a in China. AB - Whole-genome sequencing of human adenovirus type 11 (HAdV-11) strain QS, isolated in China, was conducted, and its sequence was compared with the sequences of strains within the species of HAdVs. The HAdV-11 QS genome contains 34,755 nucleotides. Similar to the other HAdV subgenus B sequences, the HAdV-11 QS genome coded 37 functional proteins and could be divided into four early, two intermediate, and five late transcription regions. The amino acid sequences of the fiber and the hypervariable regions (HVRs) within the hexon gene of HAdV-11 QS were identical to the corresponding sequences of the HAdV-11a strain; further analyses that compared those amino acid sequences with the amino acid sequences of the HAdV species subgenus B:2 strains revealed that the highest degree of homology (>99.2%) existed between HAdV-11 QS and the prototypical HAdV-14 strain, except for a few coding sequences of HVRs within the hexon gene, DNA polymerase, pVI, and pre-terminal protein. This indicate that HAdV-11 strain QS, isolated in China, is a recombinant adenovirus of HAdV-14, and the recombination analyses also confirmed this finding. It is difficult to clarify the time and manner of the recombination, and further investigations are required to determine whether the emergence of recombination between HAdV-11a and HAdV-14 might increase virulence, thereby posing a new global challenge with regard to acute respiratory diseases in the near future. PMID- 19675213 TI - Fatal Actinomucor elegans var. kuwaitiensis infection following combat trauma. AB - We report the first case of invasive mucormycosis secondary to Actinomucor elegans infection. A severely injured soldier with a fatal A. elegans var. kuwaitiensis infection is described. The identification of this fungus was performed by classical and molecular methods, and this report documents the pathogenicity of the recently described variety Actinomucor elegans var. kuwaitiensis. PMID- 19675214 TI - Evaluation of a prototype real-time PCR assay for carcinogenic human papillomavirus (HPV) detection and simultaneous HPV genotype 16 (HPV16) and HPV18 genotyping. AB - Results from a prototype real-time PCR assay that separately detected human papillomavirus genotype 16 (HPV16), HPV18, and 12 other carcinogenic HPV genotypes in aggregate (cobas 4800 HPV test) and results from a PCR assay that detects 37 HPV genotypes individually (Linear Array) were compared using a convenience sample of cervical specimens (n = 531). The percentage of total agreement between the two assays was 94.7% (95% confidence interval, 92.5 to 96.5%). The Linear Array test was more likely than cobas 4800 HPV test to test positive for the 12 other carcinogenic HPV genotypes among women without evidence of cervical disease (P = 0.004). PMID- 19675215 TI - Molecular identification of Aspergillus species collected for the Transplant Associated Infection Surveillance Network. AB - A large aggregate collection of clinical isolates of aspergilli (n = 218) from transplant patients with proven or probable invasive aspergillosis was available from the Transplant-Associated Infection Surveillance Network, a 6-year prospective surveillance study. To determine the Aspergillus species distribution in this collection, isolates were subjected to comparative sequence analyses by use of the internal transcribed spacer and beta-tubulin regions. Aspergillus fumigatus was the predominant species recovered, followed by A. flavus and A. niger. Several newly described species were identified, including A. lentulus and A. calidoustus; both species had high in vitro MICs to multiple antifungal drugs. Aspergillus tubingensis, a member of the A. niger species complex, is described from clinical specimens; all A. tubingensis isolates had low in vitro MICs to antifungal drugs. PMID- 19675216 TI - Impact of a short period of pre-enrichment on detection and bacterial loads of methicillin-resistant Staphylococcus aureus from screening specimens. AB - We compared the impacts of direct plating on a chromogenic medium and of plating after enrichment (4 h and overnight) on the detection of methicillin-resistant Staphylococcus aureus (MRSA) from 52 patient screening samples. MRSA colony counts for approximately 70% of samples after overnight pre-enrichment were >20 fold higher than the counts after the other two treatments. The qualitative differences (sample was MRSA positive/negative) between the results of the direct plating and 4-h pre-enrichment treatments were not significant, indicating no advantage of the latter; however, the number of samples positive for MRSA increased significantly after an overnight sample pre-enrichment (P < 0.005). PMID- 19675217 TI - Disease presentation in relation to infection foci for non-pregnancy-associated human listeriosis in England and Wales, 2001 to 2007. AB - Listeriosis is a rare but severe food-borne disease, affecting unborn or newly delivered infants, the elderly, and the immunocompromised. The epidemiology of listeriosis in England and Wales changed between 2001 and 2007, with more patients > or = 60 years old presenting with bacteremia (but without central nervous system [CNS] involvement). In order to explain this increase and understand the altered disease presentation, clinical, microbiological, and seasonal data on bacteremic cases of Listeria monocytogenes infection identified through national surveillance were compared with those for patients with CNS infections. Logistic regression analysis was applied while controlling for age. Bacteremic patients, who presented more frequently with gastrointestinal symptoms, were more likely to have underlying medical conditions than CNS patients. This was most marked in patients with malignancies, particularly digestive organ malignancies. Treatment to reduce stomach acid secretion modified the effect of nonmalignant underlying conditions on outcome, i.e., patients with an underlying condition who were not taking acid-suppressing medication were equally likely to have a bacteremic or a CNS infection. However, this type of therapy did not modify the effect of malignancies on the likelihood of having a bacteremic or a CNS infection. The increase in the incidence of human listeriosis among patients > or = 60 years old in England and Wales between 2001 and 2007 appears to have occurred in those with cancer or other conditions whose treatment included acid-suppressing medication. Therefore, this vulnerable patient group needs specific dietary advice on avoiding risk factors for listeriosis. PMID- 19675218 TI - Streptococcus pneumoniae DNA load in blood as a marker of infection in patients with community-acquired pneumonia. AB - Direct detection of Streptococcus pneumoniae DNA in blood adds to culture results in the etiological diagnosis of patients with community-acquired pneumonia (CAP). Quantification of the amount of DNA, the bacterial DNA load (BDL), provides a measurement of DNAemia that may increase the understanding of the clinical relevance of S. pneumoniae DNA in blood. We evaluated the S. pneumoniae BDL as a diagnostic tool in adult patients with CAP. The BDL was determined in whole-blood samples collected simultaneously with blood for culture from 45 adult patients with CAP. After DNA extraction, S. pneumoniae DNA was detected with specific real time PCR amplification, and the BDL was calculated with a standard curve. PCR and microbiological results were compared, and the BDL was related to clinical and laboratory parameters. S. pneumoniae DNA was detected in 10/13 patients with positive blood cultures and in 67% of patients with microbiologically confirmed pneumococcal pneumonia. The positive predictive values of the receiver operating characteristic curves for the BDLs for pneumococcal infection (100%) and pneumococcal bacteremia (69%) were higher than those for the level of C-reactive protein (CRP; 43% and 23%, respectively) and the white blood cell count (WBC; 42% and 35%, respectively); the negative predictive values of these three parameters were in the same range (+/-90 and +/-97%, respectively). The BDL was higher in patients presenting with systemic inflammatory response syndrome and in patients with bacteremia. Positive correlations were observed for the BDL with WBC, CRP level, and length of stay. We conclude that the BDL supports the diagnosis of S. pneumoniae infection in patients with CAP and provides a putative marker of the severity of disease. PMID- 19675219 TI - Lysine gingipain (kgp) biovars of Porphyromonas gingivalis exhibit differential distribution on oral mucosal sites. AB - A predominant kgp biovar colonized subgingival sites and buccal and tongue mucosa in 45 of 56 adults in an isolated community. The presence of biovars 381, W83, and W83v, but not HG66, correlated with the Porphyromonas gingivalis load at diseased sites. Biovars W83 and W83v poorly colonized tongue and buccal mucosa. PMID- 19675220 TI - Porphyromonas gingivalis strain diversity. AB - Porphyromonas gingivalis is implicated in the etiology of chronic periodontitis. Genotyping studies suggest that genetic variability exists among P. gingivalis strains; however, the extent of variability remains unclear and regions of variability remain largely unidentified. To assess P. gingivalis strain diversity, we previously used heteroduplex analysis of the ribosomal operon intergenic spacer region (ISR) to type strains in clinical samples and identified 22 heteroduplex types. Additionally, we used ISR sequence analysis to determine the relatedness of P. gingivalis strains to one another and demonstrated a link between ISR sequence phylogeny and the disease-associated phenotype of the strains. In the current study, heteroduplex analysis of the ISR was used to determine the worldwide genetic variability and distribution of P. gingivalis, and microarray-based comparative genomic hybridization (CGH) analysis was used to more comprehensively examine the variability of major heteroduplex type strains by using the entire genome. Heteroduplex analysis of clinical samples from geographically diverse populations identified 6 predominant geographically widespread heteroduplex types (prevalence, > or = 5%) and 14 rare heteroduplex types (prevalence, <2%) which are found in one or a few locations. CGH analysis of the genomes of seven clinically prevalent heteroduplex type strains identified 133 genes from strain W83 that were divergent in at least one of the other strains. The relatedness of the strains to one another determined on the basis of genome content (microarray) analysis was highly similar to their relatedness determined on the basis of ISR sequence analysis, and a striking correlation between the genome contents and disease-associated phenotypes of the strains was observed. PMID- 19675221 TI - First case of disseminated Mycobacterium bolletii infection in a young adult patient. AB - Mycobacterium bolletii is a rapidly growing nontuberculous mycobacterium first characterized in 2006. Here, we report a case of disseminated infection caused by M. bolletii in a young adult patient. To our knowledge, this is the first case of disseminated M. bolletii infection in an otherwise healthy young adult. PMID- 19675222 TI - Immune responses of human immature dendritic cells can be modulated by the recombinant Aspergillus fumigatus antigen Aspf1. AB - Invasive aspergillosis is a significant cause of morbidity and mortality in patients after stem cell transplantation, in solid organ transplant recipients, and in patients with hematological malignancies. The interactions between human immature dendritic cells (iDCs) and Aspergillus fumigatus antigens are widely uncharacterized. We analyzed the immune response of iDCs to different recombinant A. fumigatus antigens (Aspf1 and Crf1). One of these antigens, the 18-kDa RNase Aspf1, triggered the increased level of expression of genes encoding proinflammatory cytokines and chemokines, and augmented the activation of NFkappaB and the apoptosis of iDCs. Furthermore, by fluorescence microscopy, we could demonstrate that in the first 3 h a major portion of Aspf1 accumulates on the cell surface. Finally, we could show an increased segregation of cytokines and chemokines after the stimulation of iDCs by an Aspf1 deletion mutant strain of A. fumigatus. PMID- 19675223 TI - Kinetic patterns of Candida albicans germ tube antibody in critically ill patients: influence on mortality. AB - The influence of kinetic patterns of Candida albicans germ tube antibodies (CAGTA) on mortality was analyzed in six intensive care units. Statistically significant lower mortality rates were found in patients with patterns of increasing CAGTA titers who had been treated with antifungal agents. Thus, antifungal treatment should be considered when CAGTA titers are increasing in critically ill patients. PMID- 19675224 TI - Highly efficient antiviral CD8+ T-cell induction by peptides coupled to the surfaces of liposomes. AB - In previous studies, we have demonstrated that liposomes with differential lipid components display differential adjuvant effects when antigens (Ags) are chemically coupled to their surfaces. When ovalbumin was coupled to liposomes made by using unsaturated fatty acids, it was found to be presented not only to CD4(+) T cells but also to CD8(+) T cells and induced cytotoxic T lymphocytes (CTLs) which effectively eradicated the tumor from mice. In this study, we coupled liposomes to immunodominant CTL epitope peptides derived from lymphocytic choriomeningitis virus (LCMV) and evaluated its potency as an antiviral vaccine. The intramuscular immunization of mice with the peptide-liposome conjugates along with CpG resulted in the efficient induction of antiviral CD8(+) T-cell responses which conferred complete protection against not only LCMV Armstrong but also a highly virulent mutant strain, clone 13, that establishes persistent infections in immunocompetent mice. The intranasal vaccination induced mucosal immunity effective enough to protect mice from the virus challenge via the same route. Complete protection was achieved in mice even when the Ag dose was reduced to as low as 280 ng of liposomal peptide. This form of vaccination with a single CTL epitope induced Ag-specific memory CD8(+) T cells in the absence of CD4(+) T-cell help, which could be shown by the complete protection of CD4-knockout mice in 10 weeks as well as by the analysis of recall responses. Thus, surface-linked liposomal peptide might have a potential advantage for the induction of antiviral immunity. PMID- 19675225 TI - Detection of Coccidioides antigenemia following dissociation of immune complexes. AB - Having reported that pretreatment of serum samples with EDTA at 100 degrees C improved the sensitivity for the detection of Histoplasma antigenemia, we have evaluated this method for the detection of Coccidioides antigenemia. Urine and serum samples from patients with coccidioidomycosis were tested using the MVista Coccidioides enzyme immunoassay, and serum samples with and without EDTA-heat treatment were tested. Antigenemia was detected in 28.6% of patients whose samples were not EDTA-heat treated and in 73.1% of those whose samples were treated. Antigenuria was detected in 50% of patients. Specificity of 100% was obtained in healthy subjects, but cross-reactions were seen in 22.2% of patients with histoplasmosis or blastomycosis. EDTA-heat treatment improves the sensitivity for the detection of Coccidioides antigenemia. PMID- 19675226 TI - Inhibition of the multiplication of Mycobacterium leprae by vaccination with a recombinant M. bovis BCG strain that secretes major membrane protein II in mice. AB - The ability of a recombinant Mycobacterium bovis BCG strain that secretes major membrane protein II (MMP-II) of Mycobacterium leprae (BCG-SM) to confer protection against leprosy was evaluated by use of a mouse footpad model. C57BL/6J mice intradermally inoculated with BCG-SM produced splenic T cells which secreted significant amounts of gamma interferon (IFN-gamma) in response to either the recombinant MMP-II, the M. leprae-derived membrane fraction, or the BCG-derived cytosolic fraction in vitro more efficiently than those from the mice infected with the vector control BCG strain (BCG-pMV, a BCG strain containing pMV 261). A higher percentage of CD8(+) T cells obtained from BCG-SM-inoculated mice than those obtained from BCG-pMV-inoculated mice produced intracellular IFN-gamma on restimulation with the M. leprae antigens. BCG-SM inhibited the multiplication of M. leprae in the footpads of C57BL/6J mice more efficiently than BCG-pMV. These results indicate that a BCG strain that secretes MMP-II could be a better vaccine candidate for leprosy. PMID- 19675227 TI - MeCP2 function in the basolateral amygdala in Rett syndrome. PMID- 19675228 TI - The potassium chloride cotransporter KCC-2 coordinates development of inhibitory neurotransmission and synapse structure in Caenorhabditis elegans. AB - Chloride influx through GABA-gated chloride channels, the primary mechanism by which neural activity is inhibited in the adult mammalian brain, depends on chloride gradients established by the potassium chloride cotransporter KCC2. We used a genetic screen to identify genes important for inhibition of the hermaphrodite-specific motor neurons (HSNs) that stimulate Caenorhabditis elegans egg-laying behavior and discovered mutations in a potassium chloride cotransporter, kcc-2. Functional analysis indicates that, like mammalian KCCs, C. elegans KCC-2 transports chloride, is activated by hypotonic conditions, and is inhibited by the loop diuretic furosemide. KCC-2 appears to establish chloride gradients required for the inhibitory effects of GABA-gated and serotonin-gated chloride channels on C. elegans behavior. In the absence of KCC-2, chloride gradients appear to be altered in neurons and muscles such that normally inhibitory signals become excitatory. kcc-2 is transcriptionally upregulated in the HSN neurons during synapse development. Loss of KCC-2 produces a decrease in the synaptic vesicle population within mature HSN synapses, which apparently compensates for a lack of HSN inhibition, resulting in normal egg-laying behavior. Thus, KCC-2 coordinates the development of inhibitory neurotransmission with synapse maturation to produce mature neural circuits with appropriate activity levels. PMID- 19675229 TI - Corticotropin-releasing factor receptor antagonism within the dorsal raphe nucleus reduces social anxiety-like behavior after early-life social isolation. AB - Social isolation of rats during the early part of development increases social anxiety-like behavior in adulthood. Furthermore, early-life social isolation increases the levels of corticotropin-releasing factor (CRF) receptors in the serotonergic dorsal raphe nucleus (dRN) of adult rats. Interactions between serotonin and CRF systems are thought to mediate anxiety behavior. Therefore, we investigated the effects of CRF receptor antagonism within the dRN on social anxiety-like behavior after early-life social isolation. Male rats were reared in isolation or in groups from weaning until midadolescence, and rehoused in groups and allowed to develop into adulthood. Adult rats underwent surgery to implant a drug cannula into the dRN. After recovery from surgery and acclimation to the testing arena, rats were infused with vehicle or the CRF receptor antagonist d Phe-CRF((12-41)) (50 or 500 ng) into the dRN before a social interaction test. Isolation-reared rats pretreated with vehicle exhibited increased social anxiety like behavior compared with rats reared in groups. Pretreatment of the dRN with d Phe-CRF((12-41)) significantly reduced social anxiety-like behaviors exhibited by isolation-reared rats. Overall, this study shows that early-life social stress results in heightened social anxiety-like behavior, which is reversed by CRF antagonism within the dRN. These data suggest that CRF receptor antagonists could provide a potential treatment of stress-related social anxiety. PMID- 19675230 TI - Serotonin transporter availability in the amygdala and bed nucleus of the stria terminalis predicts anxious temperament and brain glucose metabolic activity. AB - The serotonin transporter (5-HTT) plays a critical role in regulating serotonergic neurotransmission and is implicated in the pathophysiology of anxiety and affective disorders. Positron emission tomography scans using [(11)C]DASB [(11)C]-3-amino-4-(2-dimethylaminomethylphenylsulfanyl)-benzonitrile] to measure 5-HTT availability (an index of receptor density and binding) were performed in 34 rhesus monkeys in which the relationship between regional brain glucose metabolism and anxious temperament was previously established. 5-HTT availability in the amygdalohippocampal area and bed nucleus of the stria terminalis correlated positively with individual differences in a behavioral and neuroendocrine composite of anxious temperament. 5-HTT availability also correlated positively with stress-induced metabolic activity within these regions. Collectively, these findings suggest that serotonergic modulation of neuronal excitability in the neural circuitry associated with anxiety mediates the developmental risk for affect-related psychopathology. PMID- 19675231 TI - Overcoming macrophage-mediated axonal dieback following CNS injury. AB - Trauma to the adult CNS initiates multiple processes including primary and secondary axotomy, inflammation, and glial scar formation that have devastating effects on neuronal regeneration. After spinal cord injury, the infiltration of phagocytic macrophages coincides with long-distance axonal retraction from the initial site of injury, a deleterious phenomenon known as axonal dieback. We have previously shown that activated macrophages directly induce long-distance retraction of dystrophic axons in an in vitro model of the glial scar. We hypothesized that treatments that are primarily thought to increase neuronal regeneration following spinal cord injury may in fact derive a portion of their beneficial effects from inhibition of macrophage-mediated axonal retraction. We analyzed the effects of protease inhibition, substrate modification, and neuronal preconditioning on macrophage-axon interactions using our established in vitro model. General inhibition of matrix metalloproteinases and specific inhibition of MMP-9 prevented macrophage-induced axonal retraction despite significant physical interactions between the two cell types, whereas inhibition of MMP-2 had no effect. Chondroitinase ABC-mediated digestion of the aggrecan substrate also prevented macrophage-induced axonal retraction in the presence of extensive macrophage-axon interactions. The use of a conditioning lesion to stimulate intrinsic neuronal growth potential in the absence of substrate modification likewise prevented macrophage-induced axonal retraction in vitro and in vivo following spinal cord injury. These data provide valuable insight into the cellular and molecular mechanisms underlying macrophage-mediated axonal retraction and demonstrate modifications that can alleviate the detrimental effects of this unfavorable phenomenon on the postlesion CNS. PMID- 19675234 TI - Membrane-delimited coupling of TRPV1 and mGluR5 on presynaptic terminals of nociceptive neurons. AB - Transient receptor potential vanilloid subtype 1 (TRPV1) and metabotropic glutamate receptor 5 (mGluR5) located on peripheral sensory terminals have been shown to play critical roles in the transduction and modulation of pain sensation. To date, however, very little is known regarding the significance of functional expression of mGluR5 and TRPV1 on the central terminals of sensory neurons in the dorsal horn of the spinal cord. Here we show that TRPV1 on central presynaptic terminals is coupled to mGluR5 in a membrane-delimited manner, thereby contributing to the modulation of nociceptive synaptic transmission in the substantia gelatinosa neurons of the spinal cord. Further, our results demonstrate that TRPV1 is involved in the pain behaviors induced by spinal mGluR5 activation, and diacylglycerol produced by the activation of mGluR5 mediates functional coupling of mGluR5 and TRPV1 on the presynaptic terminals. Thus, mGluR5-TRPV1 coupling on the central presynaptic terminals of nociceptive neurons may be an important mechanism underlying central sensitization under pathological pain conditions. PMID- 19675232 TI - BC1 regulation of metabotropic glutamate receptor-mediated neuronal excitability. AB - Regulatory RNAs have been suggested to contribute to the control of gene expression in eukaryotes. Brain cytoplasmic (BC) RNAs are regulatory RNAs that control translation initiation. We now report that neuronal BC1 RNA plays an instrumental role in the protein-synthesis-dependent implementation of neuronal excitation-repression equilibria. BC1 repression counter-regulates translational stimulation resulting from synaptic activation of group I metabotropic glutamate receptors (mGluRs). Absence of BC1 RNA precipitates plasticity dysregulation in the form of neuronal hyperexcitability, elicited by group I mGluR-stimulated translation and signaled through the mitogen-activated protein kinase kinase/extracellular signal-regulated kinase pathway. Dysregulation of group I mGluR function in the absence of BC1 RNA gives rise to abnormal brain function. Cortical EEG recordings from freely moving BC1(-/-) animals show that group I mGluR-mediated oscillations in the gamma frequency range are significantly elevated. When subjected to sensory stimulation, these animals display an acute group I mGluR-dependent propensity for convulsive seizures. Inadequate RNA control in neurons is thus causally linked to heightened group I mGluR-stimulated translation, neuronal hyperexcitability, heightened gamma band oscillations, and epileptogenesis. These data highlight the significance of small RNA control in neuronal plasticity. PMID- 19675233 TI - Visual processing in the central bee brain. AB - Visual scenes comprise enormous amounts of information from which nervous systems extract behaviorally relevant cues. In most model systems, little is known about the transformation of visual information as it occurs along visual pathways. We examined how visual information is transformed physiologically as it is communicated from the eye to higher-order brain centers using bumblebees, which are known for their visual capabilities. We recorded intracellularly in vivo from 30 neurons in the central bumblebee brain (the lateral protocerebrum) and compared these neurons to 132 neurons from more distal areas along the visual pathway, namely the medulla and the lobula. In these three brain regions (medulla, lobula, and central brain), we examined correlations between the neurons' branching patterns and their responses primarily to color, but also to motion stimuli. Visual neurons projecting to the anterior central brain were generally color sensitive, while neurons projecting to the posterior central brain were predominantly motion sensitive. The temporal response properties differed significantly between these areas, with an increase in spike time precision across trials and a decrease in average reliable spiking as visual information processing progressed from the periphery to the central brain. These data suggest that neurons along the visual pathway to the central brain not only are segregated with regard to the physical features of the stimuli (e.g., color and motion), but also differ in the way they encode stimuli, possibly to allow for efficient parallel processing to occur. PMID- 19675235 TI - Enigmatic central canal contacting cells: immature neurons in "standby mode"? AB - The region that surrounds the central canal of the spinal cord derives from the neural tube and retains a substantial degree of plasticity. In turtles, this region is a neurogenic niche where newborn neurons coexist with precursors, a fact that may be related with the endogenous repair capabilities of low vertebrates. Immunohistochemical evidence suggests that the ependyma of the mammalian spinal cord may contain cells with similar properties, but their actual nature remains unsolved. Here, we combined immunohistochemistry for cell-specific markers with patch-clamp recordings to test the hypothesis that the ependyma of neonatal rats contains immature neurons similar to those in low vertebrates. We found that a subclass of cells expressed HuC/D neuronal proteins, doublecortin, and PSA-NCAM (polysialylated neural cell adhesion molecule) but did not express NeuN (anti-neuronal nuclei). These immature neurons displayed electrophysiological properties ranging from slow Ca(2+)-mediated responses to fast repetitive Na(+) spikes, suggesting different stages of maturation. These cells originated in the embryo, because we found colocalization of neuronal markers with 5-bromo-2'-deoxyuridine when injected during embryonic day 7-17 but not in postnatal day 0-5. Our findings represent the first evidence that the ependyma of the rat spinal cord contains cells with molecular and functional features similar to immature neurons in adult neurogenic niches. The fact that these cells retain the expression of molecules that participate in migration and neuronal differentiation raises the possibility that the ependyma of the rat spinal cord is a reservoir of immature neurons in "standby mode," which under some circumstances (e.g., injury) may complete their maturation to integrate spinal circuits. PMID- 19675236 TI - Spikes and membrane potential oscillations in hair cells generate periodic afferent activity in the frog sacculus. AB - To look for membrane potential oscillations that may contribute to sensory coding or amplification in the ear, we made whole-cell and perforated-patch recordings from hair cells and postsynaptic afferent neurites in the explanted frog sacculus, with mechanoelectrical transduction (MET) blocked. Small depolarizing holding currents, which may serve to replace the in vivo resting MET current, evoked all-or-none calcium spikes (39-75 mV amplitude) in 37% of hair cells tested, and continuous membrane potential oscillations (14-28 mV; 15-130 Hz) in an additional 14% of cells. Spiking hair cells were on average taller and thinner than nonspiking hair cells, and had smaller outward currents through delayed rectifier channels (I(KV)) and noninactivating calcium-activated potassium channels (I(BK,steady)), and larger inward rectifier currents (I(K1)). Some spiking hair cells fired only a brief train at the onset of a current step, but others could sustain repetitive firing (3-70 Hz). Partial blockade of I(BK) changed the amplitude and frequency of oscillations and spikes, and converted some nonspiking cells into spiking cells. Oscillatory hair cells preferentially amplified sinusoidal stimuli at frequencies near their natural oscillation frequency. Postsynaptic recordings revealed regularly timed bursts of EPSPs in some afferent neurites. EPSP bursts were able to trigger afferent spikes, which may be initiated at the sodium channel cluster located adjacent to the afferent axon's most peripheral myelin segment. These results show that some frog saccular hair cells can generate spontaneous rhythmic activity that may drive periodic background activity in afferent axons. PMID- 19675237 TI - TRPC Channels Mediate a Muscarinic Receptor-Induced Afterdepolarization in Cerebral Cortex. AB - Activation of muscarinic cholinergic receptors on pyramidal cells of the cerebral cortex induces the appearance of a slow afterdepolarization that can sustain autonomous spiking after a brief excitatory stimulus. Accordingly, this phenomenon has been hypothesized to allow for the transient storage of memory traces in neuronal networks. Here we investigated the molecular basis underlying the muscarinic receptor-induced afterdepolarization using molecular biological and electrophysiological strategies. We find that the ability of muscarinic receptors to induce the inward aftercurrent underlying the slow afterdepolarization is inhibited by expression of a Galpha(q-11) dominant negative and is also markedly reduced in a phospholipase C beta1 (PLCbeta1) knock out mouse. Furthermore, we show, using a genetically encoded biosensor, that activation of muscarinic receptor induces the breakdown of phosphatidylinositol 4,5-bisphosphate in pyramidal cells. These results indicate that the Galpha(q 11)/PLCbeta1 cascade plays a key role in the ability of muscarinic receptors to signal the inward aftercurrent. We have shown previously that the muscarinic afterdepolarization is mediated by a calcium-activated nonselective cation current, suggesting the possible involvement of TRPC channels. We find that expression of a TRPC dominant negative inhibits, and overexpression of wild-type TRPC5 or TRPC6 enhances, the amplitude of the muscarinic receptor-induced inward aftercurrent. Furthermore, we find that coexpression of TRPC5 and T-type calcium channels is sufficient to reconstitute a muscarinic receptor-activated inward aftercurrent in human embryonic kidney HEK-293 cells. These results indicate that TRPC channels mediate the muscarinic receptor-induced slow afterdepolarization seen in pyramidal cells of the cerebral cortex and suggest a possible role for TRPC channels in mnemonic processes. PMID- 19675239 TI - Best1 is a gene regulated by nerve injury and required for Ca2+-activated Cl- current expression in axotomized sensory neurons. AB - We investigated the molecular determinants of Ca(2+)-activated chloride current (CaCC) expressed in adult sensory neurons after a nerve injury. Dorsal root ganglia express the transcripts of three gene families known to induce CaCCs in heterologous systems: bestrophin, tweety, and TMEM16. We found with quantitative transcriptional analysis and in situ hybridization that nerve injury induced upregulation of solely bestrophin-1 transcripts in sensory neurons. Gene screening with RNA interference in single neurons demonstrated that mouse Best1 is required for the expression of CaCC in injured sensory neurons. Transfecting injured sensory neurons with bestrophin-1 mutants inhibited endogenous CaCC. Exogenous expression of the fusion protein green fluorescent protein-Bestrophin-1 in naive neurons demonstrated a plasma membrane localization of the protein that generates a CaCC with biophysical and pharmacological properties similar to endogenous CaCC. Our data suggest that Best1 belongs to a group of genes upregulated by nerve injury and supports functional CaCC expression in injured sensory neurons. PMID- 19675238 TI - Endothelin-1 regulates oligodendrocyte development. AB - In the postnatal brain, oligodendrocyte progenitor cells (OPCs) arise from the subventricular zone (SVZ) and migrate into the developing white matter, where they differentiate into oligodendrocytes and myelinate axons. The mechanisms regulating OPC migration and differentiation are not fully defined. The present study demonstrates that endothelin-1 (ET-1) is an astrocyte-derived signal that regulates OPC migration and differentiation. OPCs in vivo and in culture express functional ET(A) and ET(B) receptors, which mediate ET-1-induced ERK (extracellular signal-regulated kinase) and CREB (cAMP response element-binding protein) phosphorylation. ET-1 exerts both chemotactic and chemokinetic effects on OPCs to enhance cell migration; it also prevents lineage progression from the O4(+) to the O1(+) stage without affecting cell proliferation. Astrocyte conditioned medium stimulates OPC migration in culture through ET receptor activation, whereas multiphoton time-lapse imaging shows that selective ET receptor antagonists or anti-ET-1 antibodies inhibit OPC migration from the SVZ. Inhibition of ET receptor activity also derepresses OPC differentiation in the corpus callosum in slice cultures. Our findings indicate that ET-1 is a soluble astrocyte-derived signal that regulates OPC migration and differentiation during development. PMID- 19675240 TI - A New Transgenic Mouse Model of Gerstmann-Straussler-Scheinker Syndrome Caused by the A117V Mutation of PRNP. AB - Gerstmann-Straussler-Scheinker syndrome (GSS) is a genetic prion disease typified clinically by the development of progressive ataxia and dementia, and histopathologically by the presence of prion protein (PrP) amyloid plaques in the CNS, especially within the cerebellum. Several mutations of the PrP gene (PRNP) are associated with GSS, but only the P102L mutation has been convincingly modeled in transgenic (Tg) mice. To determine whether other mutations carry specific GSS phenotypic information, we constructed Tg mice that express PrP carrying the mouse homolog of the GSS-associated A117V mutation. Tg(A116V) mice express approximately six times the endogenous levels of PrP, develop progressive ataxia by approximately 140 d, and die by approximately 170 d. Compared with a mouse model of transmissible Creutzfeldt-Jakob disease (CJD), the ataxia of Tg(A116V) mice is more prominent, and the course of disease is more protracted, paralleling that observed in human disease. Neuropathology includes mild scattered vacuolation and prominent, mainly cerebellar localized, thioflavin S positive PrP plaques comprised of full-length PrP(A116V). In some mice, more prominent vacuolation or a noncerebellar distribution of PrP plaques was evident, suggesting some variability in phenotype. The biophysical properties of PrP from Tg(A116V) mice and human GSS(A117V) revealed a similarly low fraction of insoluble PrP and a weakly protease-resistant approximately 13 kDa midspan PrP fragment, not observed in CJD. Overall, Tg(A116V) mice recapitulate many clinicopathologic features of GSS(A117V) that are distinct from CJD, supporting PrP(A116V) to carry specific phenotypic information. The occasional variation in histopathology they exhibit may shed light on a similar observation in human GSS(A117V). PMID- 19675241 TI - Neuronal circuit remodeling in the contralateral cortical hemisphere during functional recovery from cerebral infarction. AB - Recent advances in functional imaging of human brain activity in stroke patients, e.g., functional magnetic resonance imaging, have revealed that cortical hemisphere contralateral to the infarction plays an important role in the recovery process. However, underlying mechanisms occurring in contralateral hemisphere during functional recovery have not been elucidated. We experimentally induced a complete infarction of somatosensory cortex in right hemisphere of mice and examined the neuronal changes in contralateral (left) somatosensory cortex during recovery. Both basal and ipsilateral somatosensory stimuli-evoked neuronal activity in left (intact) hemisphere transiently increased 2 d after stroke, followed by an increase in the turnover rate of usually stable mushroom-type synaptic spines at 1 week, observed by using two-photon imaging in vivo. At 4 weeks after stroke, when functional recovery had occurred, a new pattern of electrical circuit activity in response to somatosensory stimuli was established in intact ipsilateral hemisphere. Thus, the left somatosensory cortex can compensate for the loss of the right somatosensory cortex by remodeling neuronal circuits and establishing new sensory processing. This finding could contribute to establish the effective clinical treatments targeted on the intact hemisphere for the recovery of impaired functions and to achieve better quality of life of patients. PMID- 19675242 TI - Shift from hippocampal to neocortical centered retrieval network with consolidation. AB - The standard model of system-level consolidation posits that the hippocampus is part of a retrieval network for recent memories. According to this theory, the memories are gradually transferred to neocortical circuits with consolidation, where the connections within this circuit grow stronger and reorganized so that redundant and/or contextual details may be lost. Thus, remote memories are based on neocortical networks and can be retrieved independently of the hippocampus. To test this model, we measured regional brain activity and connectivity during retrieval with functional magnetic resonance imaging. Subjects were trained on two sets of face-location association and were tested with two different delays, 15 min and 24 h including a whole night of sleep. We hypothesized that memory traces of the locations associated with specific faces will be linked through the hippocampus for the retrieval of recently learned association, but with consolidation, the activity and the functional connectivity between the neocortical areas will increase. We show that posterior hippocampal activity related to high-confidence retrieval decreased and neocortical activity increased with consolidation. Moreover, the connectivity between the hippocampus and the neocortical regions decreased and in turn, cortico-cortical connectivity between the representational areas increased. The results provide mechanistic support for a two-level process of the declarative memory system, involving initial representation of new associations in a network including the hippocampus and subsequent consolidation into a predominantly neocortical network. PMID- 19675245 TI - Stressed memories: how acute stress affects memory formation in humans. AB - Stressful, aversive events are extremely well remembered. Such a declarative memory enhancement is evidently beneficial for survival, but the same mechanism may become maladaptive and culminate in mental diseases such as posttraumatic stress disorder (PTSD). Stress hormones are known to enhance postlearning consolidation of aversive memories but are also thought to have immediate effects on attentional, sensory, and mnemonic processes at memory formation. Despite their significance for our understanding of the etiology of stress-related mental disorders, effects of acute stress at memory formation, and their brain correlates at the system scale, remain elusive. Using an integrated experimental approach, we probed the neural correlates of memory formation while participants underwent a controlled stress induction procedure in a crossover design. Physiological (cortisol level, heart rate, and pupil dilation) and subjective measures confirmed acute stress. Remarkably, reduced hippocampal activation during encoding predicted stress-enhanced memory performance, both within and between participants. Stress, moreover, amplified early visual and inferior temporal responses, suggesting that hypervigilant processing goes along with enhanced inferior temporal information reduction to relay a higher proportion of task-relevant information to the hippocampus. Thus, acute stress affects neural correlates of memory formation in an unexpected manner, the understanding of which may elucidate mechanisms underlying psychological trauma etiology. PMID- 19675244 TI - Glycinergic projection neurons of the cerebellum. AB - The cerebellum funnels its entire output through a small number of presumed glutamatergic premotor projection neurons in the deep cerebellar nuclei and GABAergic neurons that feed back to the inferior olive. Here we use transgenic mice selectively expressing green fluorescent protein in glycinergic neurons to demonstrate that many premotor output neurons in the medial cerebellar (fastigial) nuclei are in fact glycinergic, not glutamatergic as previously thought. These neurons exhibit similar firing properties as neighboring glutamatergic neurons and receive direct input from both Purkinje cells and excitatory fibers. Glycinergic fastigial neurons make functional projections to vestibular and reticular neurons in the ipsilateral brainstem, whereas their glutamatergic counterparts project contralaterally. Together, these data suggest that the cerebellum can influence motor outputs via two distinct and complementary pathways. PMID- 19675246 TI - Attention improves object representation in visual cortical field potentials. AB - Selective attention improves perception and modulates neuronal responses, but how attention-dependent changes of cortical activity improve the processing of attended objects is an open question. Changes in total signal strength or enhancements in signal-to-noise ratio have been proposed as putative mechanisms. However, it is still not clear whether, and to what extent, these processes contribute to the large perceptual improvements. We studied the ability to discriminate states of activity in visual cortex evoked by differently shaped objects depending on selective attention in monkeys. We found that gamma-band activity from V4 and V1 contains a high amount of information about stimulus shape, which increases for V4 recordings considerably with attention in successful trials, but not in case of behavioral errors. This effect resulted from enhanced differences between the stimulus-specific distributions of power spectral amplitudes. It could be explained neither by enhancements of signal-to noise ratios, nor by changes in total signal power. Instead our results indicate that attention causes underlying cortical network states to become more distinct for different stimuli, providing a new neurophysiological explanation for improvements of behavioral performance by attention. The absence of the enhancement in discriminability in trials with behavioral errors demonstrates the relevance of this novel neural mechanism for perception. PMID- 19675243 TI - Layer II/III of the prefrontal cortex: Inhibition by the serotonin 5-HT1A receptor in development and stress. AB - The modulation of the prefrontal cortex by the neurotransmitter serotonin (5-HT) is thought to play a key role in determining adult anxiety levels. Layer II/III of the prefrontal cortex, which mediates communication across cortical regions, displays a high level of 5-HT(1A) receptor binding in normal individuals and a significantly lower level in patients with mood and anxiety disorders. Here, we examine how serotonin modulates pyramidal neurons in layer II/III of the rat prefrontal cortex throughout postnatal development and in adulthood. Using whole cell recordings in brain slices of the rat medial prefrontal cortex, we observed that serotonin directly inhibits layer II/III pyramidal neurons through 5-HT(1A) receptors across postnatal development (postnatal days 6-96). In adulthood, a sex difference in these currents emerges, consistent with human imaging studies of 5 HT(1A) receptor binding. We examined the effects of early life stress on the 5 HT(1A) receptor currents in layer II/III. Surprisingly, animals subjected to early life stress displayed significantly larger 5-HT(1A)-mediated outward currents throughout the third and fourth postnatal weeks after elevated 5-HT(1A) expression during the second postnatal week. Subsequent exposure to social isolation in adulthood resulted in the almost-complete elimination of 5-HT(1A) currents in layer II/III neurons suggesting an interaction between early life events and adult experiences. These data represent the first examination of functional 5-HT(1A) receptors in layer II/III of the prefrontal cortex during normal development as well as after stress. PMID- 19675247 TI - TrkB modulates fear learning and amygdalar synaptic plasticity by specific docking sites. AB - Understanding the modulation of the neural circuitry of fear is clearly one of the most important aims in neurobiology. Protein phosphorylation in response to external stimuli is considered a major mechanism underlying dynamic changes in neural circuitry. TrkB (Ntrk2) neurotrophin receptor tyrosine kinase potently modulates synaptic plasticity and activates signal transduction pathways mainly through two phosphorylation sites [Y515/Shc site; Y816/PLCgamma (phospholipase Cgamma) site]. To identify the molecular pathways required for fear learning and amygdalar synaptic plasticity downstream of TrkB, we used highly defined genetic mouse models carrying single point mutations at one of these two sites (Y515F or Y816F) to examine the physiological relevance of pathways activated through these sites for pavlovian fear conditioning (FC), as well as for synaptic plasticity as measured by field recordings obtained from neurons of different amygdala nuclei. We show that a Y816F point mutation impairs acquisition of FC, amygdalar synaptic plasticity, and CaMKII signaling at synapses. In contrast, a Y515F point mutation affects consolidation but not acquisition of FC to tone, and also alters AKT signaling. Thus, TrkB receptors modulate specific phases of fear learning and amygdalar synaptic plasticity through two main phosphorylation docking sites. PMID- 19675248 TI - Age-dependent impairment of spine morphology and synaptic plasticity in hippocampal CA1 neurons of a presenilin 1 transgenic mouse model of Alzheimer's disease. AB - Presenilin 1 (PS1) mutations are responsible for a majority of early onset familial Alzheimer's disease (FAD) cases, in part by increasing the production of Abeta peptides. However, emerging evidence suggests other possible effects of PS1 on synaptic dysfunction where PS1 might contribute to the pathology independent of Abeta. We chose to study the L286V mutation, an aggressive FAD mutation which has never been analyzed at the electrophysiological and morphological levels. In addition, we analyzed for the first time the long term effects of wild-type human PS1 overexpression. We investigated the consequences of the overexpression of either wild-type human PS1 (hPS1) or the L286V mutated PS1 variant (mutPS1) on synaptic functions by analyzing synaptic plasticity and associated spine density changes from 3 to 15 months of age. We found that mutPS1 induces a transient increase observed only in 4- to 5-month-old mutPS1 animals in NMDA receptor (NMDA R)-mediated responses and LTP compared with hPS1 mice and nontransgenic littermates. The increase in synaptic functions is concomitant with an increase in spine density. With increasing age, however, we found that the overexpression of human wild-type PS1 progressively decreased NMDA-R-mediated synaptic transmission and LTP, without neurodegeneration. These results identify for the first time a transient increase in synaptic function associated with L286V mutated PS1 variant in an age-dependent manner. In addition, they support the view that the PS1 overexpression promotes synaptic dysfunction in an Abeta independent manner and underline the crucial role of PS1 during both normal and pathological aging. PMID- 19675249 TI - Evidence of mirror neurons in human inferior frontal gyrus. AB - There is much current debate about the existence of mirror neurons in humans. To identify mirror neurons in the inferior frontal gyrus (IFG) of humans, we used a repetition suppression paradigm while measuring neural activity with functional magnetic resonance imaging. Subjects either executed or observed a series of actions. Here we show that in the IFG, responses were suppressed both when an executed action was followed by the same rather than a different observed action and when an observed action was followed by the same rather than a different executed action. This pattern of responses is consistent with that predicted by mirror neurons and is evidence of mirror neurons in the human IFG. PMID- 19675250 TI - The geometry of perisaccadic visual perception. AB - Our ability to explore our surroundings requires a combination of high-resolution vision and frequent rotations of the visual axis toward objects of interest. Such gaze shifts are themselves a source of powerful retinal stimulation, and so the visual system appears to have evolved mechanisms to maintain perceptual stability during movements of the eyes in space. The mechanisms underlying this perceptual stability can be probed in the laboratory by briefly presenting a stimulus around the time of a saccadic eye movement and asking subjects to report its position. Under such conditions, there is a systematic misperception of the probes toward the saccade end point. This perisaccadic compression of visual space has been the subject of much research, but few studies have attempted to relate it to specific brain mechanisms. Here, we show that the magnitude of perceptual compression for a wide variety of probe stimuli and saccade amplitudes is quantitatively predicted by a simple heuristic model based on the geometry of retinotopic representations in the primate brain. Specifically, we propose that perisaccadic compression is determined by the distance between the probe and saccade end point on a map that has a logarithmic representation of visual space, similar to those found in numerous cortical and subcortical visual structures. Under this assumption, the psychophysical data on perisaccadic compression can be appreciated intuitively by imagining that, around the time of a saccade, the brain confounds nearby oculomotor and sensory signals while attempting to localize the position of objects in visual space. PMID- 19675252 TI - What does it mean to be a family physician?: Exploratory study with family medicine residents from 3 countries. AB - OBJECTIVE: To explore the conceptions that family medicine residents from 3 countries have of the roles and responsibilities of family physicians in order to gain a better understanding of challenges that might transcend the specific contexts of different health care systems. DESIGN: Qualitative study using focus groups. SETTING: Resident training programs in France, Belgium, and Canada. PARTICIPANTS: A total of 57 residents in the last year of training. METHOD: Ten focus groups were conducted in 3 countries: 2 in France, 3 in Belgium, and 5 in Canada. All focus groups were held in different cities, with residents registered in different universities in France and Canada and with residents from the same university in Belgium. The study was informed by Abbott's conceptual framework on the system of professions. Each 90-minute focus group was moderated by the same researchers. The transcripts were analyzed according to the immersion crystallization method. MAIN FINDINGS: Respondents shared common conceptions of the family physician's role: continuity of care and patient advocacy were seen as the foundations of the discipline. Respondents also shared a sense of discomfort about how accessible they were expected to be for patients and about the scope of family practice. They saw family medicine as flexible and reported that they strove for balance between their professional and personal life goals. All respondents strongly believed that their profession was undervalued by the medical schools where they trained. CONCLUSION: This exploratory study suggests that there are more similarities than differences in the understanding that future family physicians from different countries have of their discipline and of their careers. We observed a tension between a desire to develop a "new general practice" and the more traditional vision of the discipline. The culture in academic settings appears to contribute to the persistent low appeal of being a primary care physician. PMID- 19675251 TI - Functional connectivity delineates distinct roles of the inferior frontal cortex and presupplementary motor area in stop signal inhibition. AB - The neural basis of motor response inhibition has drawn considerable attention in recent imaging literature. Many studies have used the go/no-go or stop signal task to examine the neural processes underlying motor response inhibition. In particular, showing greater activity during no-go (stop) compared with go trials and during stop success compared with stop error trials, the right inferior prefrontal cortex (IFC) has been suggested by numerous studies as the cortical area mediating response inhibition. Many of these same studies as well as others have also implicated the presupplementary motor area (preSMA) in this process, in accord with a function of the medial prefrontal cortex in goal-directed action. Here we used connectivity analyses to delineate the roles of IFC and preSMA during stop signal inhibition. Specifically, we hypothesized that, as an integral part of the ventral attention system, the IFC responds to a stop signal and expedites the stop process in the preSMA, the primary site of motor response inhibition. This hypothesis predicted that preSMA and primary motor cortex would show functional interconnectivity via the basal ganglia circuitry to mediate response execution or inhibition, whereas the IFC would influence the basal ganglia circuitry via connectivity with preSMA. The results of Granger causality analyses in 57 participants confirmed this hypothesis. Furthermore, psychophysiological interaction showed that, compared with stop errors, stop successes evoked greater effective connectivity between the IFC and preSMA, providing additional support for this hypothesis. These new findings provided evidence critically differentiating the roles of IFC and preSMA during stop signal inhibition and have important implications for our understanding of the component processes of inhibitory control. PMID- 19675253 TI - Approaches to diversity in family medicine: "I have always tried to be colour blind". AB - OBJECTIVE: To explore family physicians' perceptions of and experiences with patient diversity, including differences in sex, race, ethnicity, social class, sexual orientation, and abilities or disabilities. DESIGN: Semistructured, in depth, qualitative interviews. SETTING Halifax metropolitan region, Nova Scotia. PARTICIPANTS: Twenty-two family physicians who ranged in age (25 to 65 years) and in years of practice (< 5 to > 20). Participants included both sexes, members of racialized minority groups, and those who self-identified as gay, lesbian, or bisexual. METHODS: Physicians were recruited through information letters distributed by mail and through professional networks. Interviews and field notes were recorded, transcribed verbatim, and coded using data analysis software. Weekly team discussions enhanced interpretation and analysis. MAIN FINDINGS: Family physicians employed 5 main approaches to diversity: maintaining that differences do not matter, accommodating sociocultural differences, seeking to better understand differences, seeking to avoid discrimination, and challenging inequities. Quotes from interviews illustrate these themes. CONCLUSION: Most approaches assume that both medicine (as a profession) and physicians are and should be socially and culturally neutral; some acknowledge that the sociocultural background of patients can raise tensions. Most participants in our study seek to treat patients as individuals in order to not stereotype, which hinders recognition of the ways in which sociocultural factors-both patients' and physicians'-influence health and health care. Critical reflexivity demands that physicians understand social relations of power and where they fit within those relations. PMID- 19675254 TI - Should doctors treat themselves or not? PMID- 19675256 TI - Mindful medical practice: just another fad? PMID- 19675257 TI - Should family physicians treat themselves or not?: Yes. PMID- 19675259 TI - Excluded detail. PMID- 19675258 TI - Should family physicians treat themselves or not?: No. PMID- 19675260 TI - Alternate level of care: Ontario addresses the long waits. PMID- 19675261 TI - Maternal history. PMID- 19675262 TI - Brief interventions for depression in primary care: a systematic review. AB - OBJECTIVE: To assess existing, brief nonpharmacologic interventions that are available for primary care physicians with minimal training in psychotherapy to use in managing depression in adult patients. DATA SOURCES: MEDLINE was searched from 1996 to 2007, EMBASE was searched from 1980 to 2007, and EBM Reviews was searched from 1999 to 2007. STUDY SELECTION: Several randomized controlled trials were selected using specified criteria. Selected articles were subsequently appraised and qualitatively analyzed. SYNTHESIS: Significant improvements on depression scales were found in 6 out of 8 studies (P < .05) using various brief interventions and formal control groups. Successful interventions included bibliotherapy, websites based on cognitive-behavioural therapy (CBT), and CBT based computer programs. Completion rates were highest when interventions were shorter, more structured, and included frequent contact or reminders from study staff. Validity limitations included small sample sizes, non-blinding of studies, and an uncertain degree of generalizability. CONCLUSION: Bibliotherapy, CBT-based websites, and CBT-based computer programs might be effective in assisting primary care physicians who have minimal training in psychotherapy in treating adult patients with depression. Health care personnel contact with patients undergoing these interventions might result in increased effectiveness. Future research is warranted in this area, and despite several limitations, findings from this study could help guide efforts in the development and evaluation of such research. PMID- 19675263 TI - Pharmacologic treatment of hyperthyroidism during lactation. AB - QUESTION: I have a patient who has hyperthyroidism due to Graves disease. She was taking methimazole but discontinued when she found out she was pregnant. She is currently close to delivery and might require antithyroid therapy in the postpartum period. Can methimazole cross into human milk, and is breastfeeding safe for her infant? ANSWER: The exposure of infants to methimazole or propylthiouracil through breast milk is minimal and not clinically significant. Women with hyperthyroidism using methimazole or propylthiouracil should not be discouraged from breastfeeding, as the benefits of breastfeeding largely outweigh the theoretical minimal risks. PMID- 19675264 TI - [Battlefield Blues: Ambivalence about treatment among military Veterans with depression]. PMID- 19675265 TI - Intensive glycemic control: implications of the accord, advance, and VADT trials for family physicians. PMID- 19675266 TI - Question: can you identify this condition? PMID- 19675267 TI - Physicians as parents: parenting experiences of physicians in Newfoundland and Labrador. AB - OBJECTIVE: To investigate the experiences of physicians as parents and to see if there were any differences in the parenting challenges perceived by male and female physicians. DESIGN: Mailed survey. SETTING: Newfoundland and Labrador. PARTICIPANTS: The survey was mailed to 180 male and 180 female licensed physicians, with a response rate of 60% (N = 216). MAIN OUTCOME MEASURES: Self reported experiences of being a parent and a physician. RESULTS: Female physicians reported spending significantly more time on child care activities and domestic activities than their male counterparts did (P < .001). There was no significant difference in the number of professional hours between the 2 sexes, but income was significantly lower for female physicians (P < .001). More women than men had positive physician-parent role models, although very few physicians of either sex had such role models. Female physicians reported bearing the most responsibility for the day-to-day functioning of the family; male physicians relied on their female partners to carry out the main family responsibilities. Women reported feeling guilty about their performance as mothers and as doctors. Male physicians reported regrets about the lack of time with family. CONCLUSION: Although women make up an increasing percentage of the physician work force in Canada, they still face challenges as they continue to take primary responsibility for child care and domestic activities. Women are torn between their careers and their families and sometimes feel inadequate in both roles. Male physicians regret having a lack of time with family. Strategies need to be employed in both the workplace and at home to achieve an acceptable balance between being a physician and being a parent. PMID- 19675268 TI - Health practices of Canadian physicians. AB - OBJECTIVE: To study the health and health practices of Canadian physicians, which can often influence patient health. DESIGN: Mailed survey. SETTING: Canada. PARTICIPANTS: A random sample of 8100 Canadian physicians; 7934 were found to be eligible and 3213 responded (40.5% response rate). MAIN OUTCOME MEASURES: Factors that influence health, such as consumption of fruits and vegetables, amount of exercise and alcohol consumption, smoking status, body mass idex, and participation in preventive health screening measures, as well as work-life balance and emotional stability. RESULTS: Canadian physicians are healthy. More than 90% reported being in good to excellent health, and only 5% reported that poor physical or mental health made it difficult to handle their workload more than half the time in the previous month (although a quarter had reduced work activity because of long-term health conditions). Eight percent were obese, 3% currently smoked cigarettes, and 1% typically consumed 5 drinks or more on days when they drank alcohol. Physicians averaged 4.7 hours of exercise per week and ate fruits and vegetables 4.8 times a day. Their personal screening practices were largely compliant with Canadian Task Force on Preventive Health Care recommendations. They averaged 38 hours per week on patient care and 11 hours on other professional activities. Fifty-seven percent agreed that they had a good work-life balance, and 11% disagreed with the statement "If I can, I work when I am ill." CONCLUSION: Compared with self-reports from the general Canadian population, Canadian physicians, like American physicians, seem to be healthy and to have generally healthy behaviour. There is, however, room for improvement in physicians' personal and professional well-being, and improving their personal health practices could be an efficient and beneficent way to improve the health of all Canadians. PMID- 19675270 TI - As good as it gets. PMID- 19675269 TI - The frozen man of Queens County: surgical and social outcomes in the 1860s. PMID- 19675271 TI - Quantitative and qualitative research: received and interpretivist views of science. PMID- 19675272 TI - Que sera, sera; whatever will be, will be... PMID- 19675273 TI - The price of research. PMID- 19675275 TI - Cardiac and skeletal muscle fatty acid transport and transporters and triacylglycerol and fatty acid oxidation in lean and Zucker diabetic fatty rats. AB - We examined fatty acid transporters, transport, and metabolism in hearts and red and white muscles of lean and insulin-resistant (week 6) and type 2 diabetic (week 24) Zucker diabetic fatty (ZDF) rats. Cardiac fatty acid transport was similar in lean and ZDF hearts at week 6 but was reduced at week 24 (-40%) in lean but not ZDF hearts. Red muscle of ZDF rats exhibited an early susceptibility to upregulation (+66%) of fatty acid transport at week 6 that was increased by 50% in lean and ZDF rats at week 24 but remained 44% greater in red muscle of ZDF rats. In white muscle, no differences were observed in fatty acid transport between groups or from week 6 to week 24. In all tissues (heart and red and white muscle), FAT/CD36 protein and plasmalemmal content paralleled the changes in fatty acid transport. Triacylglycerol content in red and white muscles, but not heart, in lean and ZDF rats correlated with fatty acid transport (r = 0.91) and sarcolemmal FAT/CD36 (r = 0.98). Red and white muscle fatty acid oxidation by isolated mitochondria was not impaired in ZDF rats but was reduced by 18-24% in red muscle of lean rats at week 24. Thus, in red, but not white, muscle of insulin-resistant and type 2 diabetic animals, a marked upregulation in fatty acid transport and intramuscular triacylglycerol was associated with increased levels of FAT/CD36 expression and plasmalemmal content. In heart, greater rates of fatty acid transport and FAT/CD36 in ZDF rats (week 24) were attributable to the inhibition of age-related reductions in these parameters. However, intramuscular triacylglycerol did not accumulate in hearts of ZDF rats. Thus insulin resistance and type 2 diabetes are accompanied by tissue-specific differences in FAT/CD36 and fatty acid transport and metabolism. Upregulation of fatty acid transport increased red muscle, but not cardiac, triacylglycerol accumulation. White muscle lipid metabolism dysregulation was not observed. PMID- 19675276 TI - Impaired exercise tolerance and skeletal muscle myopathy in sulfonylurea receptor 2 mutant mice. AB - By sensing intracellular energy levels, ATP-sensitive potassium (K(ATP)) channels help regulate vascular tone, glucose metabolism, and cardioprotection. SUR2 mutant mice lack full-length K(ATP) channels in striated and smooth muscle and display a complex phenotype of hypertension and coronary vasospasm. SUR2 mutant mice also display baseline cardioprotection and can withstand acute sympathetic stress better than normal mice. We now studied response to a form of chronic stress, namely that induced by 4 wk of daily exercise on SUR2 mutant mice. Control mice increased exercise capacity by 400% over the training period, while SUR2 mutant mice showed little increase in exercise capacity. Unexercised SUR2 mutant showed necrotic and regenerating fibers in multiple muscle skeletal muscles, including quadriceps, tibialis anterior, and diaphragm muscles. Unlike exercised control animals, SUR2 mutant mice did not lose weight, presumably due to less overall exertion. Unexercised SUR2 mutant mice showed a trend of mildly reduced cardiac function, measured by fractional shortening, (46 +/- 4% vs. 57 +/ 7% for SUR2 mutant and control, respectively), and this decrease was not exacerbated by chronic exercise exposure. Despite an improved response to acute sympathetic stress and baseline cardioprotection, exercise intolerance results from lack of SUR2 K(ATP) channels in mice. PMID- 19675277 TI - Loss of vagal anti-inflammatory effect: in vivo visualization and adoptive transfer. AB - The vagus nerve is a conduit for bidirectional signaling between the brain and the viscera. Vagal signaling has been shown to downregulate gastrointestinal inflammation, and the mechanism is thought to involve acetylcholine binding to the alpha-7 subunit of the nicotinic acetylcholine receptor on macrophages. The aims of this study were to quantify the impact of vagotomy in vivo by visualizing nuclear factor (NF)-kappaB activity and to determine if the proinflammatory impact of vagotomy could be transferred by lymphocytes. Real-time biophotonic imaging revealed that subdiaphragmatic vagotomy resulted in increased levels of NF-kappaB in vivo. NF-kappaB activation was further exaggerated in vivo following exposure to 4% DSS for 5 days. Vagotomized animals also exhibited higher disease activity scores and secreted more proinflammatory cytokines. Adoptive transfer of CD4(+) T cells from vagotomized animals (but not CD4(+) T cells from sham operated controls) to naive dextran sulfate sodium (DSS)-treated recipients resulted in increased inflammatory scores. Further examination of the CD4(+) T cells revealed that adoptive transfer of the CD25(-) population alone from vagotomized donors (but not sham-operated donors) was sufficient to aggravate colitis in DSS-treated recipients. Increased DSS-induced inflammation was associated with reduced CD4(+)CD25(+)Foxp3(+) regulatory T cell numbers in recipients. This study clearly demonstrates the ability of the vagus nerve to modulate activity of the proinflammatory transcription factor NF-kappaB in vivo. The proinflammatory effect of vagotomy is transferable using splenic T cells and highlights a previously unappreciated cellular mechanism for linking central parasympathetic processes with mucosal inflammation and immune homeostasis. PMID- 19675278 TI - Sex-specific differences in cardiac control and hematology of sockeye salmon (Oncorhynchus nerka) approaching their spawning grounds. AB - Some male salmonids (e.g., rainbow trout) display profound cardiovascular adjustments during sexual maturation, including cardiac growth and hypertension, and tachycardia has been observed in free-ranging male salmonids near their spawning grounds. In the present study, we investigated cardiac control, dorsal aortic blood pressure, cardiac morphometrics, and hematological variables in wild, sexually maturing sockeye salmon (Oncorhynchus nerka) with a particular aim to decipher any sex-specific differences. Routine heart rate (f(H)) was significantly higher in females (52 vs. 43 beats/min), which was due to significantly lower cholinergic tone (28 vs. 46%), because there were no differences in adrenergic tone or intrinsic heart rate between sexes. No differences in blood pressure were observed despite males possessing an 11% greater relative ventricular mass. Concomitant with higher routine heart rates, female sockeye had significantly higher levels of cortisol, testosterone, and 17beta-estradiol, whereas the level of 11-ketotestosterone was higher in males. There were no differences in hematocrit or hemoglobin concentration between the sexes. The findings of this study highlight the importance of considering sex as a variable in research fields such as conservation biology and when modeling the consequences of local and global climate change. Indeed, this study helps to provide a mechanistic basis for the significantly higher rates of female mortality observed in previous studies of wild-caught sockeye salmon. PMID- 19675279 TI - Effects of contraction on localization of GLUT4 and v-SNARE isoforms in rat skeletal muscle. AB - In skeletal muscle, contractions increase glucose uptake due to a translocation of GLUT4 glucose transporters from intracellular storage sites to the surface membrane. Vesicle-associated membrane proteins (VAMPs) are believed to play an important role in docking and fusion of the GLUT4 transporters at the surface membrane. However, knowledge about which VAMP isoforms colocalize with GLUT4 vesicles in mature skeletal muscle and whether they translocate during muscle contractions is incomplete. The aim of the present study was to further identify VAMP isoforms, which are associated with GLUT4 vesicles and examine which VAMP isoforms translocate to surface membranes in skeletal muscles undergoing contractions. VAMP2, VAMP3, VAMP5, and VAMP7 were enriched in immunoprecipitated GLUT4 vesicles. In response to 20 min of in situ contractions, there was a redistribution of GLUT4 (+64 +/- 13%), transferrin receptor (TfR; +75 +/- 22%), and insulin-regulated aminopeptidase (IRAP; +70 +/- 13%) to fractions enriched in heavy membranes away from low-density membranes (-32 +/- 7%; -18 +/- 12%; -33 +/- 9%; respectively), when compared with the resting contralateral muscle. Similarly, there was a redistribution of VAMP2 (+240 +/- 40%), VAMP5 (+79 +/- 9%), and VAMP7 (+79 +/- 29%), but not VAMP3, to fractions enriched in heavy membranes away from low-density membranes (-49 +/- 10%, -54 +/- 9%, -14 +/- 11%, respectively) in contracted vs. resting muscle. In summary, VAMP2, VAMP3, VAMP5, and VAMP7 coimmunoprecipitate with intracellular GLUT4 vesicles in muscle, and VAMP2, VAMP5, VAMP7, but not VAMP3, translocate to the cell surface membranes similar to GLUT4, TfR, and IRAP in response to muscle contractions. These findings suggest that VAMP2, VAMP5, and VAMP7 may be involved in translocation of GLUT4 during muscle contractions. PMID- 19675280 TI - The role of nitric oxide in the development of neurogenic pulmonary edema in spinal cord-injured rats: the effect of preventive interventions. AB - Neurogenic pulmonary edema (NPE) is an acute life-threatening complication following an injury of the spinal cord or brain, which is associated with sympathetic hyperactivity. The role of nitric oxide (NO) in NPE development in rats subjected to balloon compression of the spinal cord has not yet been examined. We, therefore, pretreated Wistar rats with the NO synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME) either acutely (just before the injury) or chronically (for 4 wk prior to the injury). Acute (but not chronic) L NAME administration enhanced NPE severity in rats anesthetized with 1.5% isoflurane, leading to the death of 83% of the animals within 10 min after injury. Pretreatment with either the ganglionic blocker pentolinium (to reduce blood pressure rise) or the muscarinic receptor blocker atropine (to lessen heart rate decrease) prevented or attenuated NPE development in these rats. We did not observe any therapeutic effects of atropine administered 2 min after spinal cord compression. Our data indicate that NPE development is dependent upon a marked decrease of heart rate under the conditions of high blood pressure elicited by the activation of the sympathetic nervous system. These hemodynamic alterations are especially pronounced in rats subjected to acute NO synthase inhibition. In conclusion, nitric oxide has a partial protective effect on NPE development because it attenuates sympathetic vasoconstriction and consequent baroreflex induced bradycardia following spinal cord injury. PMID- 19675282 TI - Attenuation of peripheral salt taste responses and local immune function contralateral to gustatory nerve injury: effects of aldosterone. AB - Dietary sodium restriction coupled with axotomy of the rat chorda tympani nerve (CTX) results in selectively attenuated taste responses to sodium salts in the contralateral, intact chorda tympani nerve. Converging evidence indicates that sodium deficiency also diminishes the activated macrophage response to injury on both the sectioned and contralateral, intact sides of the tongue. Because a sodium-restricted diet causes a robust increase in circulating aldosterone, we tested the hypothesis that changes in neurophysiological and immune responses contralateral to the CTX could be mimicked by aldosterone administration instead of the low-sodium diet. Taste responses in rats with CTX and supplemental aldosterone for 4-6 days were similar to rats with CTX and dietary sodium restriction. Responses to sodium salts were as much as 50% lower compared with sham-operated and vehicle-supplemented rats. The group-related functional differences were eliminated with lingual application of amiloride, suggesting that a major transduction pathway affected was through epithelial sodium channels. Consistent with the functional results, few macrophages were observed on either side of the tongue in rats with CTX and aldosterone. In contrast, macrophages were elevated on both sides of the tongue in rats with CTX and the vehicle. These results show that sodium deficiency or administration of aldosterone suppresses the immune response to neural injury, resulting in attenuation of peripheral gustatory function. They also show a potential key link among downstream consequences of sodium imbalance, taste function, and immune activity. PMID- 19675283 TI - Opioid growth factor-opioid growth factor receptor axis is a physiological determinant of cell proliferation in diverse human cancers. AB - The opioid growth factor (OGF) regulates cell proliferation of human cancer cells through the cyclin-dependent kinase inhibitory pathway, with mediation of this action by the OGF receptor (OGFr). The ubiquity of the OGF-OGFr axis in human cancer is unknown. We used 31 human cancer cell lines, representative of more than 90% of neoplasias occurring in humans, and found that OGF and OGFr were detected in the cytoplasm and nucleus by immunohistochemistry. The addition of OGF to cultures depressed cell number up to 41%, whereas naltrexone (NTX) increased cell proliferation by up to 44%, a total of 85% in the modulating capacity for the OGF-OGFr axis. Neutralization of OGF by specific antibodies led to a marked increase in cell number. Knockdown of OGFr by OGFr-siRNA resulted in a significant increase in the number of cells, even in the face of the addition of exogenous OGF. The cultures to which NTX was added and subjected to OGFr-siRNA were similar to those with OGF-siRNA alone. The OGF-OGFr axis, a physiological determinant of cell-proliferative activity, is a ubiquitous feature of human cancer cells. The identification of this native biological system in neoplasia may be important in understanding the pathophysiology of neoplasia, and in designing treatment modalities that utilize the body's own chemistry. PMID- 19675281 TI - Hibernation induces pentobarbital insensitivity in medulla but not cortex. AB - The 13-lined ground squirrel (Ictidomys tridecemlineatus), a hibernating species, is a natural model of physiological adoption to an extreme environment. During torpor, body temperature drops to 0-4 degrees C, and the cortex is electrically silent, yet the brain stem continues to regulate cardiorespiratory function. The mechanisms underlying selective inhibition in the brain during torpor are not known. To test whether altered GABAergic function is involved in regional and seasonal differences in neuronal activity, cortical and medullary slices from summer-active (SA) and interbout aroused (IBA) squirrels were placed in a standard in vitro recording chamber. Silicon multichannel electrodes were placed in cortex, ventral respiratory column (VRC), and nucleus tractus solitarius (NTS) to record spontaneous neuronal activity. In slices from IBA squirrels, bath applied pentobarbital sodium (300 microM) nearly abolished cortical neuronal activity, but VRC and NTS neuronal activity was unaltered. In contrast, pentobarbital sodium (300 microM) nearly abolished all spontaneous cortical, VRC, and NTS neuronal activity in slices from SA squirrels. Muscimol (20 microM; GABA(A) receptor agonist) abolished all neuronal activity in cortical and medullary slices from both IBA and SA squirrels, thereby demonstrating the presence of functional GABA(A) receptors. Pretreatment of cortical slices from IBA squirrels with bicuculline (100 microM; GABA(A) receptor antagonist) blocked pentobarbital-dependent inhibition of spontaneous neuronal activity. We hypothesize that GABA(A) receptors undergo a seasonal modification in subunit composition, such that cardiorespiratory neurons are uniquely unaffected by surges of an endogenous positive allosteric modulator. PMID- 19675284 TI - Role of mast cells and protease-activated receptor-2 in cyclooxygenase-2 expression in urothelial cells. AB - Mast cells have been shown to play a role in development and persistence of various inflammatory bladder disorders. Mast cell-derived tryptase specifically activates protease-activated receptor-2 (PAR-2), and PAR-2 is known to be involved in inflammation. We investigated whether mast cells participate in increase of cyclooxygenase-2 (COX-2) protein abundance in urothelium/suburothelium of bladders of mice subsequent to cyclophosphamide (CYP) induced bladder inflammation. We also used primary cultures of human urothelial cells to investigate cellular mechanisms underlying activation of PAR-2 resulting in increased COX-2 expression. We found that treatment of mice with CYP (150 mg/kg ip) increased COX-2 protein abundance in bladder urothelium/suburothelium 3, 6, and 24 h after CYP (P < 0.01), and increased COX-2 protein abundance was prevented by treatment of mice with the mast cell stabilizer sodium cromolyn (10 mg/kg ip) for 4 consecutive days before CYP treatment. Incubation of freshly isolated mouse urothelium/suburothelium with a selective PAR-2 agonist, 2-furoyl LIGRLO-amide (3 microM), also increased COX-2 protein abundance (P < 0.05). We further demonstrated that 2-furoyl-LIGRLO-amide (3 microM) increased COX-2 mRNA expression and protein abundance in primary cultures of human urothelial cells (P < 0.01), and the effects of PAR-2 activation were mediated primarily by the ERK1/2 MAP kinase pathway. These data indicate that there are functional interactions among mast cells, PAR-2 activation, and increased expression of COX 2 in bladder inflammation. PMID- 19675286 TI - Synchronization of GABAergic inputs to CA3 pyramidal cells precedes seizure-like event onset in juvenile rat hippocampal slices. AB - Here we address how dynamics of glutamatergic and GABAergic synaptic input to CA3 pyramidal cells contribute to spontaneous emergence and evolution of recurrent seizure-like events (SLEs) in juvenile (P10-13) rat hippocampal slices bathed in low-[Mg(2+)] artificial cerebrospinal fluid. In field potential recordings from the CA3 pyramidal layer, a short epoch of high-frequency oscillation (HFO; 400 800 Hz) was observed during the first 10 ms of SLE onset. GABAergic synaptic input currents to CA3 pyramidal cells were synchronized and coincided with HFO, whereas the glutamatergic input lagged by approximately 10 ms. If the intracellular [Cl(-)] remained unperturbed (cell-attached recordings) or was set high with whole cell electrode solution, CA3 pyramidal cell firing peaked with HFO and GABAergic input. By contrast, with low intracellular [Cl(-)], spikes of CA3 pyramidal cells lagged behind HFO and GABAergic input. This temporal arrangement of HFO, synaptic input sequence, synchrony of GABAergic currents, and pyramidal cell firing emerged gradually with preictal discharges until the SLE onset. Blockade of GABA(A) receptor-mediated currents by picrotoxin reduced the inter-SLE interval and the number of preictal discharges and did not block recurrent SLEs. Our data suggest that dynamic changes of the functional properties of GABAergic input contribute to ictogenesis and GABAergic and glutamatergic inputs are both excitatory at the instant of SLE onset. At the SLE onset GABAergic input contributes to synchronization and recruitment of pyramidal cells. We conjecture that this network state is reached by an activity-dependent shift in GABA reversal potential during the preictal phase. PMID- 19675285 TI - Coding of repetitive transients by auditory cortex on Heschl's gyrus. AB - The capacity of auditory cortex on Heschl's gyrus (HG) to encode repetitive transients was studied in human patients undergoing surgical evaluation for medically intractable epilepsy. Multicontact depth electrodes were chronically implanted in gray matter of HG. Bilaterally presented stimuli were click trains varying in rate from 4 to 200 Hz. Averaged evoked potentials (AEPs) and event related band power (ERBP), computed from responses at each of 14 recording sites, identified two auditory fields. A core field, which occupies posteromedial HG, was characterized by a robust polyphasic AEP on which could be superimposed a frequency following response (FFR). The FFR was prominent at click rates below approximately 50 Hz, decreased rapidly as click rate was increased, but could reliably be detected at click rates as high as 200 Hz. These data are strikingly similar to those obtained by others in the monkey under essentially the same stimulus conditions, indicating that mechanisms underlying temporal processing in the auditory core may be highly conserved across primate species. ERBP, which reflects increases or decreases of both phase-locked and non-phase-locked power within given frequency bands, showed stimulus-related increases in gamma band frequencies as high as 250 Hz. The AEPs recorded in a belt field anterolateral to the core were typically of low amplitude, showing little or no evidence of short latency waves or an FFR, even at the lowest click rates used. The non-phase locked component of the response extracted from the ERBP showed a robust, long latency response occurring here in response to the highest click rates in the series. PMID- 19675287 TI - On the origin of event-related potentials indexing covert attentional selection during visual search. AB - Despite nearly a century of electrophysiological studies recording extracranially from humans and intracranially from monkeys, the neural generators of nearly all human event-related potentials (ERPs) have not been definitively localized. We recorded an attention-related ERP component, known as the N2pc, simultaneously with intracranial spikes and local field potentials (LFPs) in macaques to test the hypothesis that an attentional-control structure, the frontal eye field (FEF), contributed to the generation of the macaque homologue of the N2pc (m N2pc). While macaques performed a difficult visual search task, the search target was selected earliest by spikes from single FEF neurons, later by FEF LFPs, and latest by the m-N2pc. This neurochronometric comparison provides an empirical bridge connecting macaque and human experiments and a step toward localizing the neural generator of this important attention-related ERP component. PMID- 19675288 TI - Long-lasting context dependence constrains neural encoding models in rodent auditory cortex. AB - Acoustic processing requires integration over time. We have used in vivo intracellular recording to measure neuronal integration times in anesthetized rats. Using natural sounds and other stimuli, we found that synaptic inputs to auditory cortical neurons showed a rather long context dependence, up to > or =4 s (tau approximately 1 s), even though sound-evoked excitatory and inhibitory conductances per se rarely lasted greater, similar 100 ms. Thalamic neurons showed only a much faster form of adaptation with a decay constant tau <100 ms, indicating that the long-lasting form originated from presynaptic mechanisms in the cortex, such as synaptic depression. Restricting knowledge of the stimulus history to only a few hundred milliseconds reduced the predictable response component to about half that of the optimal infinite-history model. Our results demonstrate the importance of long-range temporal effects in auditory cortex and suggest a potential neural substrate for auditory processing that requires integration over timescales of seconds or longer, such as stream segregation. PMID- 19675290 TI - Activity-dependent modulation of glutamatergic signaling in the developing rat dorsal horn by early tissue injury. AB - Tissue injury in early life can produce distinctive effects on pain processing, but little is known about the underlying neural mechanisms. Neonatal inflammation modulates excitatory synapses in spinal nociceptive circuits, but it is unclear whether this results directly from altered afferent input. Here we investigate excitatory and inhibitory synaptic transmission in the rat superficial dorsal horn following neonatal hindlimb surgical incision using in vitro patch-clamp recordings and test the effect of blocking peripheral nerve activity on the injury-evoked changes. Surgical incision through the skin and muscle of the hindlimb at postnatal day 3 (P3) or P10 selectively increased the frequency, but not amplitude, of glutamatergic miniature excitatory postsynaptic currents (mEPSCs) recorded 2-3 days after injury, without altering miniature inhibitory postsynaptic current frequency or amplitude at this time point. Meanwhile, incision at P17 failed to affect excitatory or inhibitory synaptic function at 2 3 days postinjury. The elevated mEPSC frequency was accompanied by increased inward rectification of evoked alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid receptor (AMPAR)-mediated currents, but no change in AMPAR/N-methyl-D-aspartate receptor ratios, and was followed by a persistent reduction in mEPSC frequency by 9-10 days postinjury. Prolonged blockade of primary afferent input from the time of injury was achieved by administration of bupivacaine hydroxide or tetrodotoxin to the sciatic nerve at P3. The increase in mEPSC frequency evoked by P3 incision was prevented by blocking sciatic nerve activity. These results demonstrate that increased afferent input associated with peripheral tissue injury selectively modulates excitatory synaptic drive onto developing spinal sensory neurons and that the enhanced glutamatergic signaling in the dorsal horn following neonatal surgical incision is activity dependent. PMID- 19675291 TI - Enhanced calcium buffering in F344 rat cholinergic basal forebrain neurons is associated with age-related cognitive impairment. AB - Alterations in neuronal Ca(2+) homeostasis are important determinants of age related cognitive impairment. We examined the Ca(2+) influx, buffering, and electrophysiology of basal forebrain neurons in adult, middle-aged, and aged male F344 behaviorally assessed rats. Middle-aged and aged rats were characterized as cognitively impaired or unimpaired by water maze performance relative to young cohorts. Patch-clamp experiments were conducted on neurons acutely dissociated from medial septum/nucleus of the diagonal band with post hoc identification of phenotypic marker mRNA using single-cell RT-PCR. We measured whole cell calcium and barium currents and dissected these currents using pharmacological agents. We combined Ca(2+) current recording with Ca(2+)-sensitive ratiometric microfluorimetry to measure Ca(2+) buffering. Additionally, we sought changes in neuronal firing properties using current-clamp recording. There were no age- or cognition-related changes in the amplitudes or fractional compositions of the whole cell Ca(2+) channel currents. However, Ca(2+) buffering was significantly enhanced in cholinergic neurons from aged cognitively impaired rats. Moreover, increased Ca(2+) buffering was present in middle-aged rats that were not cognitively impaired. Firing properties were largely unchanged with age or cognitive status, except for an increase in the slow afterhyperpolarization in aged cholinergic neurons, independent of cognitive status. Furthermore, acutely dissociated basal forebrain neurons in which choline acetyltransferase mRNA was detected had the electrophysiological profiles of identified cholinergic neurons. We conclude that enhanced Ca(2+) buffering by cholinergic basal forebrain neurons may be important during aging. PMID- 19675293 TI - Modulation of intrinsic spiking in spinal cord neurons. AB - The vertebrate spinal cord is equipped with a number of neuronal networks that underlie repetitive patterns of behavior as locomotion. Activity in such networks is mediated not only by intrinsic cellular properties but also by synaptic coupling. In this study, we focused on the modulation of the intrinsic activity by 5-hydroxytryptamine (5-HT, serotonin) and the cholinergic agonist muscarine in spinal cord cultures (embryonic age 14 rats). We investigated theses cultures (slices and dissociated cells) at the network level using multielectrode arrays (MEAs) and at the cellular level using whole cell patch clamp. All cultures showed bursting network activity and intrinsic activity when gamma-aminobutyric acid, glycine, and glutamate transmission was blocked. Using MEAs, we observed an increase of the intrinsic activity in the ventral part of the slices with 5-HT and muscarine. In single-cell recordings we found that 43 and 35% of the cells that were silent in the absence of fast synaptic activity were transformed into intrinsically spiking cells by 5-HT and muscarine, respectively. We tested the hypothesis that these neuromodulators act via modulation of the persistent sodium currents (I(NaP)) in these neurons. We found that 5-HT increased threefold the amplitude of I(NaP), specifically in the nonintrinsically spiking cells, and thus switched these cells into intrinsically spiking cells via activation of 5-HT(2) receptor and the phospholipase C pathway. In contrast, the effect of muscarine on nonintrinsically spiking neurons seems to be independent of I(NaP). We conclude from these findings that serotoninergic and cholinergic modulation can turn silent into spontaneously spiking neurons and thus initiate new sources of activity for rhythm generation in spinal networks. PMID- 19675294 TI - Response properties of fixation neurons and their location in the frontal eye field in the monkey. AB - Electrical stimulation of the frontal eye field (FEF) has recently been reported to suppress the generation of saccades, which supports the idea that the FEF plays a role in maintaining attentive fixation. This study analyzed the activity of fixation neurons that discharged during fixation in the FEF in relation to visual fixation and saccades in trained monkeys. The neural activity of fixation neurons increased at the start of fixation and was maintained during fixation. When a fixation spot of light disappeared during steady fixation, different fixation neurons exhibited different categories of response, ranging from a decrease in activity to an increase in activity, indicating that there is a continuum of fixation neurons, from neurons with foveal visual-related activity to neurons with activity that is related to the motor act of fixating. Fixation neurons usually showed a decrease in their firing rate before the onset of visually guided saccades (Vsacs) and memory-guided saccades in any direction. The reduction in activity of fixation neurons nearly coincided with, or occurred slightly before, the increase in the activity of saccade-related movement neurons in the FEF in the same monkey. Although fixation neurons were scattered in the FEF, about two thirds of fixation neurons were concentrated in a localized area in the FEF at which electrical stimulation induced strong suppression of the initiation of Vsacs bilaterally. These results suggest that fixation neurons in the FEF are part of a suppression mechanism that could control the maintenance of fixation and the initiation of saccades. PMID- 19675292 TI - SK channels gate information processing in vivo by regulating an intrinsic bursting mechanism seen in vitro. AB - Understanding the mechanistic substrates of neural computations that lead to behavior remains a fundamental problem in neuroscience. In particular, the contributions of intrinsic neural properties such as burst firing and dendritic morphology to the processing of behaviorally relevant sensory input have received much interest recently. Pyramidal cells within the electrosensory lateral line lobe of weakly electric fish display an intrinsic bursting mechanism that relies on somato-dendritic interactions when recorded in vitro: backpropagating somatic action potentials trigger dendritic action potentials that lead to a depolarizing afterpotential (DAP) at the soma. We recorded intracellularly from these neurons in vivo and found firing patterns that were quite different from those seen in vitro: we found no evidence for DAPs as each somatic action potential was followed by a pronounced afterhyperpolarization (AHP). Calcium chelators injected in vivo reduced the AHP, thereby unmasking the DAP and inducing in vitro-like bursting in pyramidal cells. These bursting dynamics significantly reduced the cell's ability to encode the detailed time course of sensory input. We performed additional in vivo pharmacological manipulations and mathematical modeling to show that calcium influx through N-methyl-d-aspartate (NMDA) receptors activate dendritic small conductance (SK) calcium-activated potassium channels, which causes an AHP that counteracts the DAP and leads to early termination of the burst. Our results show that ion channels located in dendrites can have a profound influence on the processing of sensory input by neurons in vivo through the modulation of an intrinsic bursting mechanism. PMID- 19675295 TI - Recruitment in retractor bulbi muscle during eyeblink conditioning: EMG analysis and common-drive model. AB - To analyze properly the role of the cerebellum in classical conditioning of the eyeblink and nictitating membrane (NM) response, the control of conditioned response dynamics must be better understood. Previous studies have suggested that the control signal is linearly related to the CR as a result of recruitment within the accessory abducens motoneuron pool, which acts to linearize retractor bulbi muscle and NM response mechanics. Here we investigate possible recruitment mechanisms. Data came from simultaneous recordings of NM position and multiunit electromyographic (EMG) activity from the retractor bulbi muscle of rabbits during eyeblink conditioning, in which tone and periocular shock act as conditional and unconditional stimuli, respectively. Action potentials (spikes) were extracted and classified by amplitude. Firing rates of spikes with different amplitudes were analyzed with respect to NM response temporal profiles and total EMG spike firing rate. Four main regularities were revealed and quantified: 1) spike amplitude increased with response amplitude; 2) smaller spikes always appeared before larger spikes; 3) subsequent firing rates covaried for spikes of different amplitude, with smaller spikes always firing at higher rates than larger ones; and 4) firing-rate profiles were approximately Gaussian for all amplitudes. These regularities suggest that recruitment does take place in the retractor bulbi muscle during conditioned NM responses and that all motoneurons receive the same command signal (common-drive hypothesis). To test this hypothesis, a model of the motoneuron pool was constructed in which motoneurons had a range of intrinsic thresholds distributed exponentially, with threshold linearly related to EMG spike amplitude. Each neuron received the same input signal as required by the common-drive assumption. This simple model reproduced the main features of the data, suggesting that conditioned NM responses are controlled by a common-drive mechanism that enables simple commands to determine response topography in a linear fashion. PMID- 19675296 TI - Anatomical and electrophysiological comparison of CA1 pyramidal neurons of the rat and mouse. AB - The study of learning and memory at the single-neuron level has relied on the use of many animal models, most notably rodents. Although many physiological and anatomical studies have been carried out in rats, the advent of genetically engineered mice has necessitated the comparison of new results in mice to established results from rats. Here we compare fundamental physiological and morphological properties and create three-dimensional compartmental models of identified hippocampal CA1 pyramidal neurons of one strain of rat, Sprague Dawley, and two strains of mice, C57BL/6 and 129/SvEv. We report several differences in neuronal physiology and anatomy among the three animal groups, the most notable being that neurons of the 129/SvEv mice, but not the C57BL/6 mice, have higher input resistance, lower dendritic surface area, and smaller spines than those of rats. A surprising species-specific difference in membrane resonance indicates that both mouse strains have lower levels of the hyperpolarization-activated nonspecific cation current I(h). Simulations suggest that differences in I(h) kinetics rather than maximal conductance account for the lower resonance. Our findings indicate that comparisons of data obtained across strains or species will need to account for these and potentially other physiological and anatomical differences. PMID- 19675298 TI - Ranolazine antagonizes the effects of increased late sodium current on intracellular calcium cycling in rat isolated intact heart. AB - Pathological conditions, including ischemia and heart failure, are associated with altered sodium channel function and increased late sodium current (I(Na,L)), leading to prolonged action potential duration, increased intracellular sodium and calcium concentrations, and arrhythmias. We used anemone toxin (ATX)-II to study the effects of increasing I(Na,L) on intracellular calcium cycling in rat isolated hearts. Cardiac contraction was abolished using paralytic agents. Ranolazine (RAN) was used to inhibit late I(Na). Hearts were loaded with fluo-4 acetoxymethyl ester, and myocyte intracellular calcium transients (CaTs) were measured using laser scanning confocal microscopy. ATX (1 nM) prolonged CaT duration at 50% recovery in hearts paced at a basal rate of 2 Hz and increased the sensitivity of the heart to the development of calcium alternans caused by fast pacing. ATX increased the time required for recovery of CaT amplitude following a previous beat, and ATX induced spontaneous calcium release waves during rapid pacing of the heart. ATX prolonged the duration of repolarization from the initiation of the activation to terminal repolarization in the pseudo electrocardiogram. All actions of ATX were both reversed and prevented by subsequent or prior exposure, respectively, of hearts to RAN (10 microM). Most importantly, the increased vulnerability of the heart to the development of calcium alternans during rapid pacing was reversed or prevented by 10 microM RAN. These results suggest that enhancement of I(Na,L) alters calcium cycling. Reduction by RAN of I(Na,L)-induced dysregulation of calcium cycling could contribute to the antiarrhythmic actions of this agent in both reentrant and triggered arrhythmias. PMID- 19675297 TI - Endogenous calcium buffering capacity of substantia nigral dopamine neurons. AB - Dopamine (DA)-containing cells from the substantia nigra pars compacta (SNc) play a major role in the initiation of movement. Loss of these cells results in Parkinson's disease (PD). Changes in intracellular calcium ion concentration ([Ca(2+)](i)) elicit several events in DA cells, including spike afterhyperpolarizations (AHPs) and subthreshold oscillations underlying autonomous firing. Continuous Ca(2+) load due to Ca(2+)-dependent rhythmicity has been proposed to cause the death of DA cells in PD and normal aging. Because of the physiological and pathophysiological importance of [Ca(2+)](i) in DA cells, we characterized their intrinsic Ca(2+)-buffering capacity (K(S)) in brain slices. We introduced a fluorescent Ca(2+)-sensitive exogenous buffer (200 microM fura-2) and cells were tracked from break-in until steady state by stimulating with a single action potential (AP) every 30 s and measuring the Ca(2+) transient from the proximal dendrite. DA neurons filled exponentially with a tau of about 5 6 min. [Ca(2+)](i) was assumed to equilibrate between the endogenous Ca(2+) buffer and the exogenous Ca(2+) indicator buffer. Intrinsic buffering was estimated by extrapolating from the linear relationships between the amplitude or time constant of the Ca(2+) transients versus [fura-2]. Extrapolated Ca(2+) transients in the absence of fura-2 had mean peak amplitudes of 293.7 +/- 65.3 nM and tau = 124 +/- 13 ms (postnatal day 13 [P13] to P17 animals). Intrinsic buffering increased with age in DA neurons. For cells from animals P13-P17, K(S) was estimated to be about 110 (n = 20). In older animals (P25-P32), the estimate was about 179 (n = 10). These relatively low values may reflect the need for rapid Ca(2+) signaling, e.g., to allow activation of sK channels, which shape autonomous oscillations and burst firing. Low intrinsic buffering may also make DA cells vulnerable to Ca(2+)-dependent pathology. PMID- 19675299 TI - Measurement of resting energy expenditure in healthy children. AB - BACKGROUND: The role that the components of energy expenditure play in the etiology of childhood obesity has highlighted the need for greater accuracy and standardized protocols for the measurement of resting energy expenditure (REE). However, protocols used to assess REE in children are varied, and consensus on a suitable method for measuring REE in children has not been reached. This study was undertaken to determine the effect of measurement time and measurement device (mask or mouthpiece) on REE in healthy children. DESIGN: Following a 12-hour fast and abstinence from exercise, 23 children (age, 7-12 years) completed two 35 minute protocols: one with a face mask and the other with a mouthpiece/noseclip. Energy expenditure was measured continuously via indirect calorimetry, while device acceptability was assessed using a 6-point comfort rating scale. RESULTS: Repeated measures ANOVA indicated that there was no significant difference in REE when measured after 10, 15, 20, or 25 minutes of rest compared to 30 minutes for either the mask or mouthpiece/noseclip (REE range, 1371-1460 kcal/d). Examination of the percentage coefficient of varia tion (CV) in energy expenditure for each time period by device showed that the least variation existed after 20 minutes of measurement using the mask (CV 6%). Paired t test analysis indicated significantly less discomfort when wearing the mask compared to the mouthpiece/noseclip. CONCLUSION: It would appear that a 20-minute protocol using a mask may increase compliance and prove to be a more practical protocol for measuring REE in children. PMID- 19675300 TI - Enteral refeeding rapidly restores PN-induced reduction of hepatic mononuclear cell number through recovery of small intestine and portal vein blood flows. AB - BACKGROUND: Absence of enteral nutrition (EN) reduces hepatic mononuclear cell (MNC) numbers and impairs their functions. However, enteral refeeding (ER) for as little as 12 hours following parenteral nutrition (PN) rapidly restores hepatic MNC numbers. We hypothesized that changes in small intestine and portal vein blood flows related to feeding route might be responsible for this phenomenon. METHODS: In experiment 1, mice (n = 19) were randomized to Chow (n = 5), PN (n = 7) or ER (n = 7) groups. The Chow group was given chow ad libitum with intravenous (IV) saline for 5 days. The PN group was fed parenterally for 5 days, while the ER group was re-fed with chow for 12 hours following 5 days of PN. Then, small intestine and portal vein blood flows were monitored and hepatic MNCs were isolated and counted. In experiment 2, the effects of intravenous administration of prostaglandin E(1) (PGE(1)) on hepatic MNC numbers were examined in fasted mice for 12 hours. Mice (n = 28) were randomized to Control (n = 8), PG0 (n = 10), or PG1 (n = 10) groups. The Control group was fed chow ad libitum with IV saline, while the PG0 and PG1 groups were fasted for 12 hours with infusions, respectively, of saline and PGE(1) at 1 microg/kg/minute. Blood flows and hepatic MNC numbers were examined. RESULTS: Experiment 1: ER restored PN-induced reductions in small intestine and portal vein blood flows and hepatic MNC number to the levels in the Chow group. Small intestine and portal vein blood flows correlated positively with hepatic MNC number. Experiment 2: Fasting decreased small intestine and portal vein blood flows and hepatic MNC number. However, PGE(1) restored portal vein blood flow to the level of the Control group, and moderately increased hepatic MNC number. There was a positive correlation between portal blood flow and hepatic MNC number. CONCLUSIONS: Reduced small intestine and portal vein blood flows may contribute to impaired hepatic immunity in the absence of EN. ER quickly restores hepatic MNC number through recovery of blood flow in both the small intestine and the portal vein. PMID- 19675301 TI - Intravascular embolization of venous catheter--causes, clinical signs, and management: a systematic review. AB - BACKGROUND: Intravascular embolization of device fragments is a rare but potentially serious complication. METHOD: A systematic search of the PubMed and MEDLINE databases for all articles pertaining to central catheter related embolization published in English between 1985 and 2007 was made. RESULTS: A total of 215 cases of intravenous catheter embolization were identified. There were 143 totally implanted venous devices (TIVD) or port catheters and 72 percutaneous venous catheters (PVC). Sites of catheter fragments following embolization were the superior vena cava or peripheral veins (15.4%), the right atrium (27.6%), right ventricle (22%), and pulmonary arteries (35%). Clinical signs of catheter embolization included catheter malfunction (56.3%), arrhythmia (13%), pulmonary symptoms (4.7%), and septic syndromes (1.8%), but 24.2% of cases were asymptomatic. The causes of intravascular catheter embolization were pinch off syndrome (40.9%), catheter injury during explantation (17.7%), catheter disconnection (10.7%), and catheter rupture (11.6%). In 19.1% of cases, the cause of catheter embolization could not be identified. Most embolized catheter fragments (93.5%) were removed percutaneously. In 4.2% of cases, fragments were retained in the vascular bed; in 2.3%, embolized fragments were removed surgically via thoracotomy. CONCLUSION: Intravascular catheter embolization can go undiagnosed for prolonged periods. Patients might be asymptomatic or may develop severe systemic clinical signs. The mortality rate is 1.8%. There were no significant differences in clinical features of embolization between TIVD and PVC groups. PMID- 19675302 TI - Secretory state regulates Zn2+ transport in gastric parietal cell of the rabbit. AB - Secretory compartments of neurons, endocrine cells, and exocrine glands are acidic and contain high levels of labile Zn2+. Previously, we reported evidence that acidity is regulated, in part, by the content of Zn2+ in the secretory [i.e., tubulovesicle (TV)] compartment of the acid-secreting gastric parietal cell. Here we report studies focusing on the mechanisms of Zn2+ transport by the TV compartment in the mammalian (rabbit) gastric parietal cell. Uptake of Zn2+ by isolated TV structures was monitored with a novel application of the fluorescent Zn2+ reporter N-(6-methoxy-8-quinolyl)-para-toluenesulfonamide (TSQ). Uptake was suppressed by removal of external ATP or blockade of H+-K+-ATPase that mediates luminal acid secretion. Uptake was diminished with dissipation of the proton gradient across the TV membrane, suggesting Zn2+/H+ antiport as the connection between Zn2+ uptake and acidity in the TV lumen. In isolated gastric glands loaded with the reporter fluozin-3, inhibition of H+-K+-ATPase arrested the flow of Zn(2+) from the cytoplasm to the TV compartment and secretory stimulation with forskolin enhanced vectorial movement of cytoplasmic Zn2+ into the tubulovesicle/lumen (TV/L) compartment. Our findings suggest that Zn2+ accumulation in the TV/L compartment is physiologically coupled to secretion of acid. These findings offer novel insight into mechanisms regulating Zn2+ homeostasis in the gastric parietal cell and potentially other cells in which acidic subcellular compartments serve signature functional roles. PMID- 19675303 TI - Orai1, a critical component of store-operated Ca2+ entry, is functionally associated with Na+/Ca2+ exchanger and plasma membrane Ca2+ pump in proliferating human arterial myocytes. AB - Ca(2+) entry through store-operated channels (SOCs) in the plasma membrane plays an important role in regulation of vascular smooth muscle contraction, tone, and cell proliferation. The C-type transient receptor potential (TRPC) channels have been proposed as major candidates for SOCs in vascular smooth muscle. Recently, two families of transmembrane proteins, Orai [also known as Ca(2+) release activated Ca(2+) channel modulator (CRACM)] and stromal interacting molecule 1 (STIM1), were shown to be essential for the activation of SOCs mainly in nonexcitable cells. Here, using small interfering RNA, we show that Orai1 plays an essential role in activating store-operated Ca(2+) entry (SOCE) in primary cultured proliferating human aortic smooth muscle cells (hASMCs), whereas Orai2 and Orai3 do not contribute to SOCE. Knockdown of Orai1 protein expression significantly attenuated SOCE. Moreover, inhibition of Orai1 downregulated expression of Na(+)/Ca(2+) exchanger type 1 (NCX1) and plasma membrane Ca(2+) pump isoform 1 (PMCA1). The rate of cytosolic free Ca(2+) concentration decay after Ca(2+) transients in Ca(2+)-free medium was also greatly decreased under these conditions. This reduction of Ca(2+) extrusion, presumably via NCX1 and PMCA1, may be a compensation for the reduced SOCE. Immunocytochemical observations indicate that Orai1 and NCX1 are clustered in plasma membrane microdomains. Cell proliferation was attenuated in hASMCs with disrupted Orai1 expression and reduced SOCE. Thus Orai1 appears to be a critical component of SOCE in proliferating vascular smooth muscle cells, and may therefore be a key player during vascular growth and remodeling. PMID- 19675304 TI - Vitamin K promotes mineralization, osteoblast-to-osteocyte transition, and an anticatabolic phenotype by {gamma}-carboxylation-dependent and -independent mechanisms. AB - The vitamin K family members phylloquinone (vitamin K1) and the menaquinones (vitamin K2) are under study for their roles in bone metabolism and as potential therapeutic agents for skeletal diseases. We have investigated the effects of two naturally occurring homologs, phytonadione (vitamin K1) and menatetrenone (vitamin K2), and those of the synthetic vitamin K, menadione (vitamin K3), on human primary osteoblasts. All homologs promoted in vitro mineralization by these cells. Vitamin K1-induced mineralization was highly sensitive to warfarin, whereas that induced by vitamins K2 and K3 was less sensitive, implying that gamma-carboxylation and other mechanisms, possibly genomic actions through activation of the steroid xenobiotic receptor, are involved in the effect. The positive effect on mineralization was associated with decreased matrix synthesis, evidenced by a decrease from control in expression of type I collagen mRNA, implying a maturational effect. Incubation in the presence of vitamin K2 or K3 in a three-dimensional type I collagen gel culture system resulted in increased numbers of cells with elongated cytoplasmic processes resembling osteocytes. This effect was not warfarin sensitive. Addition of calcein to vitamin K-treated cells revealed vitamin K-dependent deposition of mineral associated with cell processes. These effects are consistent with vitamin K promoting the osteoblast to-osteocyte transition in humans. To test whether vitamin K may also act on mature osteocytes, we tested the effects of vitamin K on MLO-Y4 cells. Vitamin K reduced receptor activator of NF-kappaB ligand expression relative to osteoprotegerin by MLO-Y4 cells, an effect also seen in human cultures. Together, our findings suggest that vitamin K promotes the osteoblast-to-osteocyte transition, at the same time decreasing the osteoclastogenic potential of these cells. These may be mechanisms by which vitamin K optimizes bone formation and integrity in vivo and may help explain the net positive effect of vitamin K on bone formation. PMID- 19675305 TI - Closed-state inactivation in Kv4.3 isoforms is differentially modulated by protein kinase C. AB - Kv4.3, with its complex open- and closed-state inactivation (CSI) characteristics, is a primary contributor to early cardiac repolarization. The two alternatively spliced forms, Kv4.3-short (Kv4.3-S) and Kv4.3-long (Kv4.3-L), differ by the presence of a 19-amino acid insert downstream from the sixth transmembrane segment. The isoforms are similar kinetically; however, the longer form has a unique PKC phosphorylation site. To test the possibility that inactivation is differentially regulated by phosphorylation, we expressed the Kv4.3 isoforms in Xenopus oocytes and examined changes in their inactivation properties after stimulation of PKC activity. Whereas there was no difference in open-state inactivation, there were profound differences in CSI. In Kv4.3-S, PMA reduced the magnitude of CSI by 24% after 14.4 s at -50 mV. In contrast, the magnitude of CSI in Kv4.3-L increased by 25% under the same conditions. Mutation of a putatively phosphorylated threonine (T504) to aspartic acid within a PKC consensus recognition sequence unique to Kv4.3-L eliminated the PMA response. The change in CSI was independent of the intervention used to increase PKC activity; identical results were obtained with either PMA or injected purified PKC. Our previously published 11-state model closely simulated our experimental data. Our data demonstrate isoform-specific regulation of CSI by PKC in Kv4.3 and show that the carboxy terminus of Kv4.3 plays an important role in regulation of CSI. PMID- 19675307 TI - Enhanced deposition of low-molecular-weight hyaluronan in lungs of cigarette smoke-exposed mice. AB - Chronic obstructive pulmonary disease (COPD) is characterized by infiltration of inflammatory cells, destruction of lung parenchyma, and airway wall remodeling. Hyaluronan (HA) is a component of the extracellular matrix, and low-molecular weight (LMW) HA fragments have proinflammatory capacities. We evaluated the presence of HA in alveolar and airway walls of C57BL/6 mice that were exposed to air or cigarette smoke (CS) for 4 weeks (subacute) or 24 weeks (chronic). We measured deposition of the extracellular matrix proteins collagen and fibronectin in airway walls and determined the molecular weight of HA purified from lung tissue. In addition, we studied the expression of HA-modulating genes by RT-PCR. HA staining in alveolar walls was significantly enhanced upon chronic CS exposure, whereas HA levels in the airway walls were already significantly higher upon subacute CS exposure and remained elevated upon chronic CS exposure. This differed from the deposition of collagen and fibronectin, which are only elevated at the chronic time point. In lungs of CS-exposed mice, the molecular weight of HA clearly shifted toward more LMW HA fragments. CS exposure significantly increased the mRNA expression of the HA synthase gene Has3 in total lung tissue, whereas the expression of Has1 was decreased. These in vivo studies in an experimental model of COPD show that CS exposure leads to enhanced deposition of (mostly LMW) HA in alveolar and bronchial walls by altering the expression of HA modulating enzymes. This may contribute to airway wall remodeling and pulmonary inflammation in COPD. PMID- 19675306 TI - Conditional deletion of dnaic1 in a murine model of primary ciliary dyskinesia causes chronic rhinosinusitis. AB - Studies of primary ciliary dyskinesia (PCD) have been hampered by the lack of a suitable animal model because disruption of essential ciliary genes in mice results in a high incidence of lethal hydrocephalus. To develop a viable mouse model for long-term studies of PCD, we have generated a transgenic mouse line in which two conserved exons of the mouse intermediate dynein chain gene, Dnaic1, are flanked by loxP sites (Dnaic1(flox/flox)). Dnaic1 is the murine homolog of human DNAI1, which is mutated in approximately 10% of human PCD cases. These mice have been crossed with mice expressing a tamoxifen-inducible Cre recombinase (CreER). Treatment of adult Dnaic1(flox/flox)/CreER(+/-) mice with tamoxifen results in an almost complete deletion of Dnaic1 with no evidence of hydrocephalus. Treated animals have reduced levels of full-length Dnaic1 mRNA, and electron micrographs of cilia demonstrate a loss of outer dynein arm structures. In treated Dnaic1(flox/flox)/CreER(+/-) animals, mucociliary clearance (MCC) was reduced over time. After approximately 3 months, no MCC was observed in the nasopharynx, whereas in the trachea, MCC was observed for up to 6 months, likely reflecting a difference in the turnover of ciliated cells in these tissues. All treated animals developed severe rhinosinusitis, demonstrating the importance of MCC to the health of the upper airways. However, no evidence of lung disease was observed up to 11 months after Dnaic1 deletion, suggesting that other mechanisms are able to compensate for the lack of MCC in the lower airways of mice. This model will be useful for the study of the pathogenesis and treatment of PCD. PMID- 19675308 TI - No kidding: high risk of cognitive difficulty in new-onset pediatric epilepsy. PMID- 19675310 TI - Using descriptive assessment in the treatment of bite acceptance and food refusal. AB - The feeding behaviors of two children who maintained failure to thrive diagnoses and displayed food refusal are assessed in their homes. Descriptive assessments are used to identify schedules of consequence provided by each child's care providers for bite acceptance and food refusal behaviors. Assessments reveal rich schedules of praise and access to social interaction and preferred activities for bite acceptance and escape for food refusal. These schedule arrangements result in hypotheses that modifications to the schedule of praise and access to social interaction and preferred activities for bite acceptance would result in little to no effect and that modifications to the schedule of escape for food refusal would be necessary for treatment success. Successful interventions are subsequently implemented by manipulating the existing schedules of escape for food refusal by each child's care providers. Implications for the use of descriptive assessments for feeding problems are discussed. PMID- 19675309 TI - Neuropsychological status at seizure onset in children: risk factors for early cognitive deficits. AB - OBJECTIVE: This large, prospective, community-based study characterized neuropsychological functioning and academic achievement at the time of the first recognized seizure and identified risk factors for cognitive deficits. METHODS: We compared 282 children (ages 6-14 years, IQ > or =70) with a first recognized seizure to 147 healthy siblings on a battery of well-standardized and widely used neuropsychological and academic achievement tests and examined relationships with demographic and clinical variables. RESULTS: In this intellectually normal cohort, 27% with just one seizure and up to 40% of those with risk factors exhibited neuropsychological deficits at or near onset. Risk factors associated with neuropsychological deficits included multiple seizures (i.e., second unprovoked seizure; odds ratio [OR] = 1.96), use of antiepileptic drugs (OR = 2.27), symptomatic/cryptogenic etiology (OR = 2.15), and epileptiform activity on the initial EEG (OR = 1.90); a child with all 4 risks is 3.00 times more likely than healthy siblings to experience neuropsychological deficits by the first clinic visit. Absence epilepsy carried increased odds for neuropsychological impairment (OR = 2.00). CONCLUSIONS: A subgroup of intellectually normal children with seizures showed neuropsychological deficits at onset. Academic achievement was unaffected, suggesting that there is a window early in the disorder for intervention to ameliorate the impact on school performance. Therefore, the risk factors identified here (especially if multiple risks are present) warrant swift referral for neuropsychological evaluation early in the course of the condition. PMID- 19675311 TI - Establishing quality assurance criteria for serial dilution operations on liquid handling equipment. AB - Since the advent of high-throughput screening (HTS) in the early 1990s, parallel multichannel liquid handlers have become a mainstay in every drug discovery setting. Although several peer-reviewed publications have discussed methods and criteria for stamping multiwell copies, there is very little information about establishing a standard operating procedure (SOP) for standard (microliter-level) serial dilutions of compounds used in dose-response experiments. The authors discuss the 4 main criteria any serial dilution process must pass (accuracy, precision, fold dilution, and outliers) and the process for establishing thresholds for all of these values in a compound management or biological screening laboratory. The thresholds need to be both low enough to be acceptable from a biological potency variability perspective and high enough to allow the instruments to pass the quality assurance (QA) analysis on a regular basis. In this article, the authors suggest suitable thresholds arrived at by a variety of methods, including trend analysis of QA data, survey questionnaire from the main stakeholders (screening scientists, chemists), and published criteria for single shot stamping. A mathematical analysis of the effect of threshold values on estimated XC(50)s was performed to ensure that the variability introduced by the serial dilution step is within acceptable overall variability limits. PMID- 19675312 TI - Test system for trifunctional antibodies in 3D MCTS culture. AB - The aim of the present study was to assess the feasibility of a 3D tumor cell culture model, that is, multicellular tumor spheroids (MCTSs) as an adequate model for micrometastases and therefore as a pharmacological model for efficacy testing of trifunctional therapeutic antibodies. Unlike conventional monolayer cultures, spheroids allow researchers to study parameters, such as 3D cell shape, 3D cell arrangement and microenvironment, and penetration efficiency of defense cells that may largely influence the efficacy of antibody treatment in vivo. The authors established a long-term coculture of human MCTSs with peripheral blood mononuclear cells (PBMCs) to test the anticancer effect of the trifunctional, bispecific antibody catumaxomab (anti-EpCAM x anti-CD3) or similar therapeutic molecules. The test system is accessible to various analytical methods and thus allows for characterizing multiple parameters, which can help elucidate the mode of action of immunotherapeutic anticancer treatment. For example, the novel approach enables precise, reproducible volume growth analysis of MCTSs under immunotherapeutic treatments. For evaluation of changes within individual spheroids, cryosections can be stained (e.g., for proliferating or apoptotic cells as well as infiltrating PBMCs). Molecular PCR-based assays or flow cytometric analyses allow for discrimination between different cell types, particularly leukocyte subtypes. Furthermore, MCTSs can be disaggregated to form standard monolayers for cell viability or plating efficiency experiments. For these reasons, the MCTS model is a powerful tool to analyze drug efficacy with various endpoints under highly reproducible, standardized conditions. PMID- 19675313 TI - Inhibitors of RecA activity discovered by high-throughput screening: cell permeable small molecules attenuate the SOS response in Escherichia coli. AB - The phenomenon of antibiotic resistance has created a need for the development of novel antibiotic classes with nonclassical cellular targets. Unfortunately, target-based drug discovery against proteins considered essential for in vitro bacterial viability has yielded few new therapeutic classes of antibiotics. Targeting the large proportion of genes considered nonessential that have yet to be explored by high-throughput screening, for example, RecA, can complement these efforts. Recent evidence suggests that RecA-controlled processes are responsible for tolerance to antibiotic chemotherapy and are involved in pathways that ultimately lead to full-fledged antibiotic resistance. Therefore inhibitors of RecA may serve as therapeutic adjuvants in combination chemotherapy of bacterial infectious diseases. Toward the goal of validating RecA as a novel target in the chemotherapy of bacterial infections, the authors have screened 35,780 small molecules against RecA. In total, 80 small molecules were identified as primary hits and could be clustered in 6 distinct chemotype clades. The most potent class of hits was further examined, and 1 member compound was found to inhibit RecA mediated strand exchange and prevent ciprofloxacin-induced SOS expression in Escherichia coli. This compound represents the first small molecule demonstrating an ability to inhibit the bacterial SOS response in live bacterial cell cultures. PMID- 19675314 TI - Mechanism-based inhibition: deriving K(I) and k(inact) directly from time dependent IC(50) values. AB - The potential of enzyme inhibition of a drug is frequently quantified in terms of IC(50) values. Although this is a suitable quantity for reversible inhibitors, concerns arise when dealing with irreversible or mechanism-based inhibitors (MBIs). IC(50) values of MBIs are time dependent, causing serious problems when aiming at ranking different compounds with respect to their inhibitory potential. As a consequence, most studies and ranking schemes related to MBIs rely on the inhibition constant (K(I)) and the rate of enzyme inactivation (k(inact)) rather than on IC(50) values. In this article, the authors derive a novel relation between potentially time-dependent IC(50) values and K(I), k(inact) parameters for different types of inhibition. This allows for direct estimation of K(I) and k(inact) values from time-dependent IC(50) values, even without the need of additional preincubation experiments. The application of this approach is illustrated using a fluorimetric assay to access the drug-drug interaction potential associated with new chemical entities. The approach can easily be implemented using standard software tools (e.g., XLfit) and may also be suitable for applications where mechanism-based inhibition is a desired mode of action (e.g., at particular pharmacological drug targets). PMID- 19675315 TI - Stem cells in drug discovery, tissue engineering, and regenerative medicine: emerging opportunities and challenges. AB - Stem cells, irrespective of their origin, have emerged as valuable reagents or tools in human health in the past 2 decades. Initially, a research tool to study fundamental aspects of developmental biology is now the central focus of generating transgenic animals, drug discovery, and regenerative medicine to address degenerative diseases of multiple organ systems. This is because stem cells are pluripotent or multipotent cells that can recapitulate developmental paths to repair damaged tissues. However, it is becoming clear that stem cell therapy alone may not be adequate to reverse tissue and organ damage in degenerative diseases. Existing small-molecule drugs and biologicals may be needed as "molecular adjuvants" or enhancers of stem cells administered in therapy or adult stem cells in the diseased tissues. Hence, a combination of stem cell-based, high-throughput screening and 3D tissue engineering approaches is necessary to advance the next wave of tools in preclinical drug discovery. In this review, the authors have attempted to provide a basic account of various stem cells types, as well as their biology and signaling, in the context of research in regenerative medicine. An attempt is made to link stem cells as reagents, pharmacology, and tissue engineering as converging fields of research for the next decade. PMID- 19675316 TI - Still wanted: teachers with knowledge of language. PMID- 19675317 TI - Teacher knowledge, instructional expertise, and the development of reading proficiency. AB - Teacher knowledge and instructional expertise have been found in correlational and pre- and posttest studies to be related to student reading achievement. This article summarizes data presented in this special issue and additional research to address four questions: (a) What do expert reading teachers know? (b) Why do teachers need to acquire this knowledge? (c) Do teachers believe they have this knowledge? and (d) Are teachers being adequately prepared to teach reading? Well designed studies relevant to this topic have been sparse with a noticeable lack of attention given to identifying specific causal links between teacher knowledge, teaching expertise, and student reading achievement. Until the appropriate research designs and methodologies are applied to address the question of causal effects, conclusions about the specific content that teachers must know and the instructional practices that are most beneficial in presenting this content are preliminary at best. Future studies of the effect of essential reading content knowledge must be extended beyond word-level skills to vocabulary, reading comprehension, and writing. PMID- 19675318 TI - Sample size in obesity trials: patient perspective versus current practice. AB - OBJECTIVE: To evaluate patient opinions on acceptable risks in exchange for a given degree of weight loss and their implications for sample size determination in obesity randomized clinical trials (RCTs). DESIGN: . Survey of patients entering RCTs for weight loss in a university-based clinical research setting and power calculations based on their responses. Participants. Men (n = 8) and women (n = 66) between 24 and 73 years of age with body mass indices ranging from 26.8 to 40.5 kg/m(2). Measurements. Survey responses to questions assessing the added risk of serious adverse events (SAEs) or death one is willing to assume for a given degree of weight loss. RESULTS: For 5% and 10% weight loss against risk for death per se, the mean acceptable risk tended to be about 3.5%, but the median (0.00) and mode (0.00) suggested that for most individuals, only a risk of < or = 1% would be acceptable. Figures, estimated dropout rates, and base rates of SAEs (including deaths) from recent obesity trials indicate that 1-year 2-group obesity RCTs would need tens of thousands of participants per group to have 80% power to detect risks that are meaningful to patients at the 2-tailed 0.05 alpha level. CONCLUSION: Patient education is needed to explain which risks are realistically detectable in RCTs so that patients may provide truly informed consent, or RCT standards should be modified to meet patients' implicit expectations. PMID- 19675320 TI - Satisfaction with care: a measure of quality of care in prostate cancer patients. AB - BACKGROUND: Patients' assessment of satisfaction with care, quality of care, and outcomes has become a central issue in patient-centered prostate cancer (PCa) care. We sought to analyze the association between patient-reported satisfaction with care and health-related quality of life (HRQoL) in newly diagnosed PCa patients. METHODS: Prospective cohort design was used to recruit 590 newly diagnosed PCa patients from an urban academic hospital and a VA hospital. Participants completed satisfaction with care (CSQ-8) and HRQoL (SF-36 and UCLA PCI) surveys prior to treatment and at 3, 6, 12, and 24 months' follow-up. Cross lagged analysis was used to ascertain the causal direction between satisfaction with care and HRQoL. Propensity scores were used to adjust for potential selection bias between treatment groups. Linear mixed models were used to analyze the association between satisfaction with care, process of care (treatment), and outcomes (generic and prostate-specific HRQoL) after adjusting for covariates. RESULTS: Cross-lagged correlation results are consistent with a cause-effect association between HRQoL and satisfaction with care. After controlling for clinical and demographic covariates, radical prostatectomy (RP) treatment was associated with higher satisfaction with care (odds ratio [OR], 7.9; P = 0.043). Improved generic and prostate-specific HRQoL were associated with higher satisfaction with care, after adjusting for demographic and clinical covariates. CONCLUSION: Satisfaction with care appears to be associated with process of care and outcomes of care. Assessment of satisfaction with care is useful for evaluating the quality of PCa care. Satisfaction with care is an important arena in cancer outcomes research, whose full potential remains unexploited. PMID- 19675319 TI - Increasing the detection and response to adherence problems with cardiovascular medication in primary care through computerized drug management systems: a randomized controlled trial. AB - BACKGROUND: Adherence with antihypertensive and lipid-lowering therapy is poor, resulting in an almost 2-fold increase in hospitalization. Treatment side effects, cost, and complexity are common reasons for nonadherence, and physicians are often unaware of these potentially modifiable problems. OBJECTIVE: To determine if a cardiovascular medication tracking and nonadherence alert system, incorporated into a computerized health record system, would increase drug profile review by primary care physicians, increase the likelihood of therapy change, and improve adherence with antihypertensive and lipid-lowering drugs. METHODS: There were 2293 primary care patients prescribed lipid-lowering or antihypertensive drugs who were randomized to the adherence tracking and alert system or active medication list alone to determine if the intervention increased drug profile review, changes in cardiovascular drug treatment, and refill adherence in the first 6 months. An intention to treat analysis was conducted using generalized estimating equations to account for clustering within physician. RESULTS: Overall, medication adherence was below 80% for 36.3% of patients using lipid-lowering drugs and 40.8% of patients using antihypertensives at the start of the trial. There was a significant increase in drug profile review in the intervention compared to the control group (44.5% v. 35.5%; P < 0.001), a nonsignificant increase in drug discontinuations due to side effects (2.3% v. 2.0%; P = 0.61), and a reduction in therapy increases (28.5% v. 29.1%; P = 0.86). There was no significant change in refill adherence after 6 months of follow-up. CONCLUSION: An adherence tracking and alert system increases drug review but not therapy changes or adherence in prevalent users of cardiovascular drug treatment. Targeting incident users where adverse treatment effects are more common and combining adherence tracking and alert tools with motivational interventions provided by multidisciplinary primary care teams may improve the effectiveness of the intervention. PMID- 19675321 TI - Bayesian hierarchical models for cost-effectiveness analyses that use data from cluster randomized trials. AB - Cost-effectiveness analyses (CEA) may be undertaken alongside cluster randomized trials (CRTs) where randomization is at the level of the cluster (for example, the hospital or primary care provider) rather than the individual. Costs (and outcomes) within clusters may be correlated so that the assumption made by standard bivariate regression models, that observations are independent, is incorrect. This study develops a flexible modeling framework to acknowledge the clustering in CEA that use CRTs. The authors extend previous Bayesian bivariate models for CEA of multicenter trials to recognize the specific form of clustering in CRTs. They develop new Bayesian hierarchical models (BHMs) that allow mean costs and outcomes, and also variances, to differ across clusters. They illustrate how each model can be applied using data from a large (1732 cases, 70 primary care providers) CRT evaluating alternative interventions for reducing postnatal depression. The analyses compare cost-effectiveness estimates from BHMs with standard bivariate regression models that ignore the data hierarchy. The BHMs show high levels of cost heterogeneity across clusters (intracluster correlation coefficient, 0.17). Compared with standard regression models, the BHMs yield substantially increased uncertainty surrounding the cost-effectiveness estimates, and altered point estimates. The authors conclude that ignoring clustering can lead to incorrect inferences. The BHMs that they present offer a flexible modeling framework that can be applied more generally to CEA that use CRTs. PMID- 19675322 TI - The association between individual time preferences and health maintenance habits. AB - CONTEXT: Encouraging healthy behaviors, including disease screening, exercise, and tobacco avoidance, has been a significant focus of clinical attention in recent decades. Little is known about the association between individual preferences with respect to time play and preventive health care use and healthy lifestyles. OBJECTIVE: To determine whether rates of these health behaviors are associated with latent time preferences. DESIGN: Interval regression analysis was used to impute individual level discount rates. The difference in means for the rates of health behaviors were assessed for high vs. low to moderate discounting groups using one-factor probit models. PARTICIPANTS: The 2004 wave of the Health and Retirement Survey included in a time preferences module (1,039 respondents aged 24 to 65 years). MAIN OUTCOME MEASURES: Rates of recent mammograms, breast exams, Pap smears, prostate exams, cholesterol testing, flu shots, and dental visits, and non-smoking status. RESULTS: Respondents in the upper 20th percentile of the distribution have an average imputed annual discount rate of 0.335 (33.5%). High discount rate status is found to have a negative marginal association on the probability that respondents had recent mammogram use (-15.1%; P = 0.001), Pap smear use (-8.3%; P = 0.049), prostate examination use (-20.4%; P =0.003), dental visits (-24.8%; P = 0.001), cholesterol testing (-12.4%; P = 0.001), flu shot usage (-11.1%; P = 0.005), rates of vigorous exercise (-15.1%; P = 0.001), nonsmoking status (-10.4%; P= 0.001), and undertook all measured health habits (-7%; P = 0.001). CONCLUSIONS: Differences in underlying preferences for the present over the future may be a substantial barrier for people's propensity to adopt healthy lifestyles. PMID- 19675323 TI - Improving decision making at the end of life with video images. AB - BACKGROUND: Decision making at the end of life is frequently complex and often filled with uncertainty. We hypothesized that people with limited health literacy would have more uncertainty about end-of-life decision making than people with adequate literacy. We also hypothesized that video images would decrease uncertainty. DESIGN: . Before and after oral survey. Participants. Subjects presenting to their primary care physicians. METHODS: Subjects were asked about their preferences for end-of-life care after they heard a verbal description of advanced dementia and were asked to rate the level of their uncertainty. Subjects then viewed a video of a patient with advanced dementia and were asked again about their preferences and uncertainty. Uncertainty was measured using the Decisional Conflict Scale with score ranges from 3 (high uncertainty) to 15 (no uncertainty). Health literacy was measured using the Rapid Estimate of Adult Literacy in Medicine, and subjects were divided into 3 literacy categories: low (0-45, 6th grade and below), marginal (46- 60, 7th-8th grade), and adequate (61 66, 9th grade and above). RESULTS: A total of 146 patients completed the interview. Prior to the video, the average uncertainty scores for subjects with low, marginal, and adequate health literacy were 10.8, 12.4, and 13.5, respectively (P < 0.0001). After the video, the 3 groups had similar uncertainty about their decisions. The average uncertainty scores for subjects with low, marginal, and adequate health literacy were 13.6, 14.1, and 14.5, respectively (P = 0.046). CONCLUSIONS: Subjects with limited health literacy expressed more uncertainty about their preferences for end-of-life care than did subjects with adequate literacy. Our video decision aid improved end-of-life decision making by decreasing uncertainty regarding subjects' preferences, especially for those with limited literacy. PMID- 19675324 TI - Poverty, wealth, and access to pandemic influenza vaccines. PMID- 19675325 TI - Understanding how Americans view health care reform. PMID- 19675327 TI - Images in clinical medicine. Esophageal foreign body. PMID- 19675326 TI - New, but not improved? Incorporating comparative-effectiveness information into FDA labeling. PMID- 19675328 TI - Practicing medicine in the age of Facebook. PMID- 19675329 TI - Micrometastases or isolated tumor cells and the outcome of breast cancer. AB - BACKGROUND: The association of isolated tumor cells and micrometastases in regional lymph nodes with the clinical outcome of breast cancer is unclear. METHODS: We identified all patients in The Netherlands who underwent a sentinel node biopsy for breast cancer before 2006 and had breast cancer with favorable primary-tumor characteristics and isolated tumor cells or micrometastases in the regional lymph nodes. Patients with node-negative disease were randomly selected from the years 2000 and 2001. The primary end point was disease-free survival. RESULTS: We identified 856 patients with node-negative disease who had not received systemic adjuvant therapy (the node-negative, no-adjuvant-therapy cohort), 856 patients with isolated tumor cells or micrometastases who had not received systemic adjuvant therapy (the node-positive, no-adjuvant-therapy cohort), and 995 patients with isolated tumor cells or micrometastases who had received such treatment (the node-positive, adjuvant-therapy cohort). The median follow-up was 5.1 years. The adjusted hazard ratio for disease events among patients with isolated tumor cells who did not receive systemic therapy, as compared with women with node-negative disease, was 1.50 (95% confidence interval [CI], 1.15 to 1.94); among patients with micrometastases, the adjusted hazard ratio was 1.56 (95% CI, 1.15 to 2.13). Among patients with isolated tumor cells or micrometastases, the adjusted hazard ratio was 0.57 (95% CI, 0.45 to 0.73) in the node-positive, adjuvant-therapy cohort, as compared with the node-positive, no-adjuvant-therapy cohort. CONCLUSIONS: Isolated tumor cells or micrometastases in regional lymph nodes were associated with a reduced 5-year rate of disease free survival among women with favorable early-stage breast cancer who did not receive adjuvant therapy. In patients with isolated tumor cells or micrometastases who received adjuvant therapy, disease-free survival was improved. PMID- 19675330 TI - Weight lifting in women with breast-cancer-related lymphedema. AB - BACKGROUND: Weight lifting has generally been proscribed for women with breast cancer-related lymphedema, preventing them from obtaining the well-established health benefits of weight lifting, including increases in bone density. METHODS: We performed a randomized, controlled trial of twice-weekly progressive weight lifting involving 141 breast-cancer survivors with stable lymphedema of the arm. The primary outcome was the change in arm and hand swelling at 1 year, as measured through displaced water volume of the affected and unaffected limbs. Secondary outcomes included the incidence of exacerbations of lymphedema, number and severity of lymphedema symptoms, and muscle strength. Participants were required to wear a well-fitted compression garment while weight lifting. RESULTS: The proportion of women who had an increase of 5% or more in limb swelling was similar in the weight-lifting group (11%) and the control group (12%) (cumulative incidence ratio, 1.00; 95% confidence interval, 0.88 to 1.13). As compared with the control group, the weight-lifting group had greater improvements in self reported severity of lymphedema symptoms (P=0.03) and upper- and lower-body strength (P<0.001 for both comparisons) and a lower incidence of lymphedema exacerbations as assessed by a certified lymphedema specialist (14% vs. 29%, P=0.04). There were no serious adverse events related to the intervention. CONCLUSIONS: In breast-cancer survivors with lymphedema, slowly progressive weight lifting had no significant effect on limb swelling and resulted in a decreased incidence of exacerbations of lymphedema, reduced symptoms, and increased strength. (ClinicalTrials.gov number, NCT00194363.) PMID- 19675331 TI - Uterine fibroid embolization. PMID- 19675332 TI - Images in clinical medicine. Testicular torsion in a hydrocele. PMID- 19675333 TI - Case records of the Massachusetts General Hospital. Case 25-2009. A 36-year-old woman with hormone-receptor-positive breast cancer. PMID- 19675334 TI - A weighty matter--lifting after breast cancer. PMID- 19675335 TI - Premature release of data from clinical trials of ezetimibe. PMID- 19675336 TI - Master regulators of female fertility. PMID- 19675337 TI - Dipyridamole plus aspirin and hemodialysis graft patency. PMID- 19675338 TI - Genomewide association studies of stroke. PMID- 19675339 TI - Vaccine refusal and the risks of vaccine-preventable diseases. PMID- 19675340 TI - Case 15-2009: a man with coma after cardiac arrest. PMID- 19675341 TI - Vision 1 year after gene therapy for Leber's congenital amaurosis. PMID- 19675342 TI - Tics in childhood. PMID- 19675343 TI - Impact factor and its role in academic promotion. PMID- 19675345 TI - Managing asthma exacerbations in the emergency department: summary of the National Asthma Education And Prevention Program Expert Panel Report 3 guidelines for the management of asthma exacerbations. PMID- 19675346 TI - Noninvasive ventilation. PMID- 19675344 TI - An official ATS proceedings: asthma in the workplace: the Third Jack Pepys Workshop on Asthma in the Workplace: answered and unanswered questions. AB - The Third Jack Pepys Workshop on Asthma in the Workplace held in Montreal, Canada in May 2007 focused on emerging data, including progress over the previous three years touching on questions identified at a previous Workshop (2004). The format, based on that used in previous workshops, consisted primarily of short, thematic, structured slide presentations followed by extensive, open-ended discussion periods. The key summary content of the workshops discussions has been distilled for this account. (Expanded details of the prepared presentations in PowerPoint format can be found at: www.asthma-workplace.com.) The topics reflect an expanded scope of interest including consideration of: (1) work-related asthma (WRA), subsuming both occupational asthma (OA) and work-exacerbated asthma (WEA); although the latter condition is commoner than OA, discussion mainly focused on OA because the corpus of scientific literature is larger; (2) other related occupational airway pathologic processes, beyond WRA, including rhinitis and eosinophilic bronchitis, with focus on various methods that improve objective confirmation of these conditions; (3) the psycho-socioeconomic impact of WRA with presentation of questionnaires that assess disability due to OA; (4) development of a world-wide perspective on work-related airway disease, including the situation in countries with emerging economies where the frequency of WRA is likely similar to or even greater than that in developed countries. The overarching conclusion was that WRA and related airway conditions are underrecognized and underdiagnosed both in developed and developing countries, with a great many aspects related to personal and environmental risk, exposure, mechanisms, and assessment of impairment/disability remaining to explore to better inform primary, secondary, and tertiary disease prevention. PMID- 19675347 TI - Intubation and mechanical ventilation of the asthmatic patient in respiratory failure. PMID- 19675348 TI - Anti-inflammatory treatment after discharge home from the emergency department in adults with acute asthma. PMID- 19675349 TI - Follow-up after acute asthma episodes: what improves future outcomes? PMID- 19675353 TI - Gap-junction proteins in retinal development: new roles for the "nexus". AB - Gap-junction channels, the cytoplasmic proteins that associate with them, and the transcriptional networks that regulate them are increasingly being viewed as critical communications hubs for cell signaling in health and disease. As a result, the term "nexus," which was the original structural name for these focal intercellular links, is coming back into use with new proteomic and transcriptomic meanings. The retina is better understood than any other part of the vertebrate central nervous system in respect of its developmental patterning, its diverse neuronal types and circuits, and the emergence of its definitive structure-function correlations. Thus, studies of the junctional and nonjunctional nexus roles of gap-junction proteins in coordinating retinal development should throw useful light on cell signaling in other developing nervous tissues. PMID- 19675352 TI - Engineering proteins for custom inhibition of Ca(V) channels. AB - The influx of Ca(2+) ions through voltage-dependent calcium (Ca(V)) channels links electrical signals to physiological responses in all excitable cells. Not surprisingly, blocking Ca(V) channel activity is a powerful method to regulate the function of excitable cells, and this is exploited for both physiological and therapeutic benefit. Nevertheless, the full potential for Ca(V) channel inhibition is not being realized by currently available small-molecule blockers or second-messenger modulators due to limitations in targeting them either to defined groups of cells in an organism or to distinct subcellular regions within a single cell. Here, we review early efforts to engineer protein molecule blockers of Ca(V) channels to fill this crucial niche. This technology would greatly expand the toolbox available to physiologists studying the biology of excitable cells at the cellular and systems level. PMID- 19675355 TI - What's new about osmotic regulation of glycerophosphocholine. AB - Glycerophosphocholine is an abundant renal medullary organic osmolyte that protects renal medullary cells from the high interstitial concentrations of NaCl and urea to which they are normally exposed. We consider the metabolism of glycerophosphocholine, its osmotic regulation, and the recently discovered molecular identity of the enzymes that osmoregulate its abundance. PMID- 19675356 TI - The mammalian urine concentrating mechanism: hypotheses and uncertainties. AB - The urine concentrating mechanism of the mammalian kidney, which can produce a urine that is substantially more concentrated than blood plasma during periods of water deprivation, is one of the enduring mysteries in traditional physiology. Owing to the complex lateral and axial relationships of tubules and vessels, in both the outer and inner medulla, the urine concentrating mechanism may only be fully understood in terms of the kidney's three-dimensional functional architecture and its implications for preferential interactions among tubules and vessels. PMID- 19675357 TI - Molecular mechanisms of ischemic cerebral edema: role of electroneutral ion transport. AB - The brain achieves homeostasis of its intracellular and extracellular fluids by precisely regulating the transport of solute and water across its major cellular barriers: endothelia of the blood-brain barrier (BBB), choroid plexus epithelia, and neuroglial cell membranes. Cerebral edema, the pathological accumulation of fluid in the brain's intracellular and extracellular spaces, is a major cause of morbidity and mortality following stroke and other forms of ischemic brain injury. Until recently, mechanisms of cerebral edema formation have been obscure; consequently, its treatment has been empiric and suboptimal. Here, we provide a paradigm for understanding ischemic cerebral edema, showing that its molecular pathogenesis is a complex yet step-wise process that results largely from impaired astrocytic cell volume regulation and permeability alterations in the cerebral microvasculature, both of which arise from pathological changes in the activities of specific ion channels and transporters. Recent data has implicated the bumetanide-sensitive NKCC1, an electroneutral cotransporter expressed in astrocytes and the BBB, in cerebral edema formation in several different rodent models of stroke. Pharmacological inhibition or genetic deficiency of NKCC1 decreases ischemia-induced cell swelling, BBB breakdown, cerebral edema, and neurotoxicity. Combination pharmacological strategies that include NKCC1 as a target might thus prove beneficial for the treatment of ischemic, and potentially other types of, cerebral edema. PMID- 19675359 TI - Commentary on Curry et al.'s safer and stronger program and suggestions for future methodological research. PMID- 19675354 TI - Phosphoinositide signaling: new tools and insights. AB - Phosphoinositides constitute only a small fraction of cellular phospholipids, yet their importance in the regulation of cellular functions can hardly be overstated. The rapid metabolic response of phosphoinositides after stimulation of certain cell surface receptors was the first indication that these lipids could serve as regulatory molecules. These early observations opened research areas that ultimately clarified the plasma membrane role of phosphoinositides in Ca(2+) signaling. However, research of the last 10 years has revealed a much broader range of processes dependent on phosphoinositides. These lipids control organelle biology by regulating vesicular trafficking, and they modulate lipid distribution and metabolism more generally via their close relationship with lipid transfer proteins. Phosphoinositides also regulate ion channels, pumps, and transporters as well as both endocytic and exocytic processes. The significance of phosphoinositides found within the nucleus is still poorly understood, and a whole new research concerns the highly phosphorylated inositols that also appear to control multiple nuclear processes. The expansion of research and interest in phosphoinositides naturally created a demand for new approaches to determine where, within the cell, these lipids exert their effects. Imaging of phosphoinositide dynamics within live cells has become a standard cell biological method. These new tools not only helped us localize phosphoinositides within the cell but also taught us how tightly phosphoinositide control can be linked with distinct effector protein complexes. The recent progress allows us to understand the underlying causes of certain human diseases and design new strategies for therapeutic interventions. PMID- 19675360 TI - Commentary on development of measures of abuse among women with disabilities and the characteristics of their perpetrators. PMID- 19675361 TI - Commentary on Powers et al.'s article: "interpersonal violence and women with disabilities: an analysis of safety promoting behaviors". PMID- 19675362 TI - Situating research on safety promoting behaviors among disabled and deaf victims of interpersonal violence. PMID- 19675364 TI - Developing teen dating violence prevention strategies: formative research with middle school youth. AB - Intimate partner violence (IPV) peaks in youth and young adulthood and is associated with multiple adolescent risk behaviors and negative health outcomes. Targeting youth with prevention messages before they start dating may avert teen dating violence and subsequent adult IPV. This article discusses findings from focus groups with middle school youth to determine behaviors and beliefs regarding dating violence. To develop effective prevention messages, participants were asked questions about characteristics of middle school dating relationships, healthy relationships, relationship norms, unhealthy relationships, emotional abuse, physical abuse, sexual abuse, intervening in violent situations, and trusted sources for information about dating violence. The recommendations for prevention efforts include an emphasis on skill building, tailoring efforts for particular subgroups, and identifying innovative ways of reaching youth. PMID- 19675365 TI - Women's awareness of and discomfort with sexual assault cues: effects of alcohol consumption and relationship type. AB - Two studies examined the effects of alcohol and relationship type on women's sexual assault risk perception. Study 1 participants (N = 62) consumed a moderate alcohol dose or nonalcoholic beverage, then rated their awareness of and discomfort with sexual assault risk cues in a hypothetical encounter with a new or established dating partner. Study 2 (N = 351) compared control, placebo, low, and high alcohol dose conditions using a similar scenario. Intoxicated women reported decreased awareness of and discomfort with risk cues. An established relationship decreased discomfort ratings. Findings indicate that alcohol may increase women's sexual victimization likelihood through reduced sexual assault risk perception. PMID- 19675366 TI - The functions of gender role traditionality, ambivalent sexism, injury, and frequency of assault on domestic violence perception: a study between Japanese and American college students. AB - This study examined the mediating influence of gender-role traditionality (GRT), ambivalent sexism, and victim injury and frequency of assault on domestic violence (DV) perception differences between Japanese and American college students. As predicted, Japanese tended to minimize, blame, and excuse DV more than did Americans, and these national differences were mediated by GRT. Participants viewed the DV incident more seriously when the victim presented injury or when the incident had occurred frequently. Those high in benevolent and hostile sexism were more likely to minimize DV, whereas those high only in benevolent sexism were more likely to blame the victim. PMID- 19675368 TI - Occupational and nonoccupational postexposure prophylaxis for HIV in 2009. AB - Data supporting the efficacy of HIV postexposure prophylaxis (PEP) come largely from a small number of older studies and case reports in health care workers, studies of transmission from infected mothers to their infants, and animal studies. These data also provide support for the current recommendations regarding duration of PEP and the window of time within which PEP should be started. Although much of the available data are from experience with older 2 drug regimens, newer potent 2- and 3-drug regimens are increasingly used in occupational exposure management, and drugs with mechanisms of action targeting early events in infection (eg, entry inhibitors, integrase inhibitors) may in the future become attractive options. Nonoccupational PEP remains controversial, although its feasibility and safety have been demonstrated in a number of programs. Existing recommendations generally call for its use within 72 hours of high-risk contact with a high-risk or HIV-infected source individual. This article summarizes a presentation on PEP for HIV infection made by Raphael J. Landovitz, MD, at the IAS-USA continuing medical education course held in Los Angeles in February 2009. The original presentation is available as a Webcast at www.iasusa.org. PMID- 19675367 TI - Update on cardiovascular complications in HIV infection. AB - HIV-infected patients are at increased risk of cardiovascular disease (CVD), reflecting interaction of risk associated with host, virus, and antiretroviral therapy factors. Although traditional risk factors appear to increase risk to a similar degree in HIV-infected and HIV-uninfected persons, some risk factors (eg, smoking) may be more common in HIV-infected persons. HIV infection per se may further increase CVD risk via proatherosclerotic effects on smooth muscle cells and macrophages or by increasing inflammation. Some drugs in the protease inhibitor class are associated with increased risk, at least partly in association with adverse lipid effects. The magnitude and mechanisms of risk reported to be associated with recent use of abacavir remain undefined. This article summarizes a presentation on cardiovascular complications in HIV infection made by Judith S. Currier, MD, at an International AIDS Society-USA continuing medical education course in Los Angeles in February 2009. The original presentation is available as a Webcast at www.iasusa.org. PMID- 19675369 TI - What's new in the 2009 US guidelines for prevention and treatment of opportunistic infections among adults and adolescents with HIV? AB - Despite dramatic declines in the incidence of opportunistic infections (OIs) in the United States, they remain an important cause of morbidity and mortality for HIV-infected persons. Previously separate guidelines on the prevention of OIs and on the treatment of OIs have been combined recently into an updated single document; the present article reviews salient changes to and new information contained in this guidance. Chapters on hepatitis B virus infection and tuberculosis have been expanded substantially, and each chapter now includes information on immune reconstitution inflammatory syndrome. In addition, there is detailed discussion on the role of antiretroviral therapy in OI prevention and issues concerning the initiation of antiretroviral therapy during treatment of an acute OI. In the future, these guidelines will likely be maintained as an internet-based document to facilitate wider dissemination and more rapid updates. PMID- 19675370 TI - Influenza A (H1N1): responding to a pandemic threat. PMID- 19675371 TI - Pictorial health warnings are a must for effective tobacco control. PMID- 19675372 TI - Salt never calls itself sweet. PMID- 19675373 TI - TPMT gene polymorphisms: on the doorstep of personalized medicine. PMID- 19675374 TI - Metabolic syndrome as a marker of risk in type 2 diabetes. PMID- 19675375 TI - Diet & insulin resistance: a review & Asian Indian perspective. AB - Insulin resistance is associated with type 2 diabetes mellitus (T2DM) and cardiovascular diseases (CVD). These abnormalities have been aggravated because of imbalanced and excess nutrition in developed countries, and rapid nutritional and lifestyle transition occurring in developing countries. This review presents evidence linking dietary nutrients with insulin resistance and its metabolic correlates, and also describes these issues from a Asian Indians and South Asian perspective. Despite possible influences from genetic and perinatal factors, diet and physical activity are likely to have greater and often overriding influence in pathogenesis of the insulin resistance, the metabolic syndrome, and T2DM. In animal studies, a link has been established between dietary nutrients and insulin resistance. However, in human studies evidence is not as strong as in animals. Data suggest that dietary omega-3 polyunsaturated fatty acids (PUFAs) improve lipid profile and may have beneficial effect on insulin resistance. Dietary saturated fatty acids intake is positively associated with insulin resistance. Also, low glycaemic index foods and whole grain intake decrease insulin resistance. Importantly, high carbohydrate diets increase plasma triglycerides, cause hyperinsulinaemia and decreases low-density lipoprotein cholesterol. Among micronutrients, high magnesium and calcium intake have been reported to decrease insulin resistance. High intake of dietary carbohydrate and omega-6 PUFAs, low intake of omega-3 PUFAs and fiber, and high omega -6/omega-3 PUFAs ratio have been reported in South Asians. Our recent investigations have shown that increased dietary omega-6 PUFAs and saturated fat intake are significantly associated with fasting hyperinsulinaemia and sub-clinical inflammation, respectively. Such imbalanced diets contribute to high prevalence of insulin resistance, the metabolic syndrome and T2DM in South Asians and Asian Indians. PMID- 19675376 TI - Application of SNaPshot for analysis of thiopurine methyltransferase gene polymorphism. AB - BACKGROUND & OBJECTIVE: Mercaptopurine, azathioprine, and thioguanine, used as antineoplastic agents and immunosuppressants are catabolized by thiopurine methyltransferase (TPMT) enzyme, which exhibits genetic polymorphism. Genotyping patients and the population to which the patients belong, is important for effective treatment and reducing toxicity. There is a need for faster methods for genotyping. Hence the present study was planned to test the application of SNaPshot technique for analysis of the three common TPMT alleles: TPMT*2, TPMT*3A, and TPMT*3C in DNA from healthy Indian volunteers as well as to apply the method on cDNA samples obtained from children with acute lymphoblastic leukaemia (ALL). METHODS: A total of 120 healthy volunteers and 25 patients were analysed by multiplexed SNaPshot reaction. Genomic DNA was the template for most of the analyses, but additionally the cDNA synthesized for translocation detection was used as the template in case of patients with ALL. The results of SNaPshot reaction were confirmed by direct sequencing. RESULTS: The TPMT genotype could be reliably identified by SNaPshot analysis in multiplex reactions both in genomic DNA samples and cDNA. The overall frequency of the three common polymorphisms was observed to be 4.9 per cent, arising from heterozygosity for TPMT*3C (4.1%) and TPMT*3A (0.8%). INTERPRETATION & CONCLUSION: SNaPshot method for TPMT polymorphism analysis works faster with the potential for high throughput. By simultaneously interrogating the genotype at multiple sites, the method can provide future opportunity to multiplex, though multiplexing has not been done in the present analysis. Heterozygosity for TPMT*3C (719 A>G) was detected in 4.1 per cent of the study population and no homozygosity was observed. Our results indicated that TPMT*3C was the most common polymorphism in Indian population, while TPMT3*A, associated with the absence of catalytic activity of TPMT, was very rare. PMID- 19675377 TI - Predictors of metabolic syndrome in Asian north Indians with newly detected type 2 diabetes. AB - BACKGROUND & OBJECTIVE: The identification of metabolic syndrome (MS) among patients with type 2 diabetes (T2DM) is of great importance, since those with MS carry a cluster of cardiovascular risk factors. This study evaluates suitable criteria with high efficiency in diagnosing MS and to identify the strongest predictors of MS in newly detected type 2 diabetes individuals. METHODS: Newly detected type 2 diabetes (<6 months) patients were assessed. The MS was assessed by WHO, National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP III), modified NCEP-ATP-III and International Diabetes Federation (IDF) criteria. Receiver operating characteristics (ROC) curves of serum triglycerides, HDL, and waist circumference were created for the prediction of MS and the area under the corresponding curves (AUC) were used to evaluate the predictive efficiency of each MS parameter. Different cut points in the selected variables and the corresponding sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were estimated. RESULTS: Among the 563 newly detected T2DM individuals, the presence of MS ranged from 57 to 68 per cent according to the different criteria. The higher percentage of MS was observed in modified NCEPATP III criteria. In comparison to men, presence of MS was higher in women in all the four criteria [198 (67%) vs. 165 (62%); 209 (70%) vs. 111 (42%); 231 (78%) vs. 151 (57%); 222 (75%) vs. 129 (49%)] by modified WHO, NCEP-ATP III, modified NCEP-ATP III, and IDF, respectively. The predictive ability to diagnose MS was highest with modified NCEP-ATP III and lowest with IDF criteria. The optimal cut-off of waist circumference in men and women were 90 and 88 cm respectively. Serum triglyceride in men effectively indicated the presence of MS in newly detected T2DM individuals, whereas, in women the HDL-C was the stronger predictor of MS. INTERPRETATION & CONCLUSION: The study results show that modified NCEP-ATP III criteria predict highest occurrence of MS in newly detected T2DM patients. Elevated serum triglyceride for men and decreased serum HDL-C in women were the strongest single predictors, effectively indicating presence of MS in newly detected T2DM. PMID- 19675378 TI - Effects of resistance training on metabolic profile of adults with type 2 diabetes. AB - BACKGROUND & OBJECTIVE: The number of diabetes in India is increasing at an alarming rate. The effects of physical activity in the form of resistance training or aerobic exercises on type 2 diabetes have not been studied in Indian population. The objective of this study was to analyse the effects of eight weeks of progressive resistance training (PRT) compared with aerobic exercise (AE) on glycaemic control, metabolic profile, cardiovascular fitness parameters and general well being in adults with type 2 diabetes. METHODS: Thirty adults (14 females and 16 males mean; age 53.8 +/- 8.8 yr) with type 2 diabetes were randomly assigned to 8 wk supervised PRT (n=10) or AE (n=10) or control group (n=10). Glycosylated haemoglobin (HbA1c), lipid profile, blood pressure, heart rate, body mass index (BMI) and general well being were measured before training (i.e. 0 wk) and after 8 wk of training period. RESULTS: Plasma glycosylated haemoglobin levels decreased significantly (P<0.05) both in the PRT group (7.57 +/- 2.4% to 6.23 +/- 0.8%) and in AE group (8.11+/-0.9% to 6.66 +/- 0.9%).Total cholesterol levels decreased significantly (P<0.05) by 13.3 per cent in PRT group and by 6.1 per cent in AE group. Both exercise groups showed significantly reduction in systolic blood pressure (P<0.05). General well being improvement was much more in PRT (8.6%) as compared to AE group (2.7%). INTERPRETATION & CONCLUSION: Our findings showed that both PRT and AE were effective in improving metabolic profile of adults with type 2 diabetes but the percentage improvement in triglycerides, total cholesterol levels and general well being with PRT was more compared to AE. Further studies on a larger sample need to be done to confirm these findings. PMID- 19675379 TI - In silico characterization of genetic homology in nuclear-encoded apicoplast targeted genes between Plasmodium falciparum & P. vivax. AB - BACKGROUND & OBJECTIVE: Resistance to anti-malarial drugs by the parasites is one of the major obstacles to malaria control. The primary objective of this work was to find specific nuclear-encoded-apicoplast-targeted genes that are conserved between two different human malaria parasite species, Plasmodium falciparum and P. vivax to find a common drug/vaccine targets for both the species. METHODS: Using computational genomics, possible nuclear-encoded-apicoplast-targeted genes were identified in P. falciparum genome. With comparative genomic approaches, homologous genes were identified between the two different human malaria species, P. falciparum and P. vivax. RESULTS: Of the total 545 reported nuclear-encoded apicoplast-targeted genes in P. falciparum, we could narrow down to as less as five genes that were found to have highly conserved nucleotide stretches in P. vivax. However, two such genes were of importance, as the majority of the protein coding regions (exons) of these genes were found to be highly conserved between them. INTERPRETATION & CONCLUSION: This preliminary study shows that nuclear encoded-apicoplast-targeted genes were conserved between the two human malaria parasites and these could be targeted for developing a common drug to cure both forms of malaria. PMID- 19675380 TI - Identification of novel variants in the COL4A4 gene in Korean patients with thin basement membrane nephropathy. AB - BACKGROUND & OBJECTIVE: The alpha4 chain of the type 4 collagen family is an important component of the glomerular basement membrane (GBM) in the kidney. It is encoded by the COL4A4 gene, and mutations of this gene are known to be associated with thin basement membrane nephropathy (TBMN). To better understand the contribution of variants in the COL4A4 gene to TBMN, we investigated the sequence of the complete COL4A4 gene in 45 Korean patients with TBMN. METHODS: Genomic DNA was obtained from the peripheral blood lymphocytes. For the analysis of the COL4A4 gene, all the exons including splicing sites were amplified by PCR and screened by direct sequencing analysis. RESULTS: Eight novel COL4A4 sequence variants were found in these patients. Two of these variants, G199R and G1606E, were possibly pathogenic variants affecting the phenotype. None of these variants were observed in 286 chromosomes from normal Korean control subjects. In addition, 39 polymorphisms including 7 novel SNPs were identified in this study. INTERPRETATION & CONCLUSION: The frequency of COL4A4 mutations in Korean patients with TBMN is low and the other cases may have mutations in other genes like COL4A3. Screening of the COL4A3 gene and finding a novel causative gene for TBMN will help clarify the pathogenesis of this disorder and perhaps for distinguishing TBMN from Alport syndrome. PMID- 19675381 TI - Characterization & evolutionary analysis of human CD36 gene. AB - BACKGROUND & OBJECTIVE: Understanding evolutionary genetic details of immune system genes responsible for infectious diseases is of prime importance concerning disease pathogenecity. Considering malaria as a devastating disease in the world including India, detail evolutionary understanding on human immune system gene is essential. The primary aim of this study was to initiate work on one such gene, the human CD36 gene responsible in malaria pathogenesis. METHODS: DNA sequences of the human CD36 gene was retrieved from public domain and fine scale details were characterized. Both comparative and evolutionary analyses were performed with sequences from six other taxa (5 mammalian one avian) where CD36 homologs are present. Different statistical analyses were also performed. RESULTS: Differential distribution in number and length of exons and introns was detected in CD36 gene across seven taxa. The CpG islands were also found to be distributed unevenly across the gene and taxa. Neighbour-joining tree was constructed and it was observed that the chimpanzee and human are diverged at the CD36 gene relatively recently. The chicken, Gallus gallus was found to be diverged from rest of the taxa significantly. Also copy number variation was observed across different taxa. INTERPRETATION & CONCLUSION: Comparative genomic study of a human immune system gene CD36 show relationships among different taxa at the evolutionary level. The information can be of help to study genetic diversity in malaria endemic zones and to correlate it with malaria pathogenecity. PMID- 19675382 TI - In vitro effect of fluoroquinolones against Mycobacterium tuberculosis isolates from Agra & Kanpur region of north India. AB - BACKGROUND & OBJECTIVE: Fluoroquinolones (FQs) are important drugs used for treatment of drug resistant tuberculosis and are also now being considered as first line drugs to shorten the duration of treatment of tuberculosis (TB). In order to find out useful FQs for treatment of tuberculosis, the comparative efficacy of five FQs, namely, ofloxacin (OFL), ciprofloxacin (CIP), sparfloxacin (SPX), gatifloxacin (GAT) and levofloxacin (LEVX) was studied against Mycobacterium tuberculosis (MTB) isolates obtained from both treated and untreated patients from Agra and Kanpur regions of north India. METHODS: A total of 162 MTB isolates [including 110 MTB isolates obtained from untreated patients (Cat-I) and 52 isolates from treated patients (Cat-II)] were tested for their susceptibilities to FQs using standard minimum inhibitory concentration (MIC) method on Lowenstein-Jensen medium. RESULTS: Keeping in view the therapeutically achievable drug levels, it was found that in Cat-I 97.2 per cent (107/110) isolates were sensitive to GAT, 89 per cent (98/110) to LEVX at 1 microg/ml whereas 92.7 per cent (102/110) isolates were inhibited by OFL at 2 microg/ml and 73.6 per cent (81/110) to SPX at 0.5 microg/ml. Only 63.6 per cent (70/110) isolates were found to be sensitive to CIP at 2 microg/ml which increased to 89 per cent (98/110) at 4 microg/ml (higher than achievable peak serum level). On the other hand, among 52 isolates for Cat-II, 37 (71.2%) were found to be sensitive to GAT and 33 (63.5%) to LEVX at 1 microg/ml concentration, 28 (53.8%) to SPX at 0.5 microg/ml whereas 33 (63.5%) and 24 (46.2%) isolates were found to be sensitive to OFL and CIP at 2 microg/ml, respectively. INTERPRETATION & CONCLUSION: It appears that GAT has higher activity against MTB isolates followed by OFL, LEVX and SPX whereas CIP showed the lowest activity. GAT was also found to be the most effective FQ against multi-drug resistant (MDR) isolates both from Cat-I and Cat-II patients. Thus, except CIP, other FQs showed potential to be included in the treatment regimens of tuberculosis including MDR-TB. PMID- 19675383 TI - Incomplete immunological recovery following anti-tuberculosis treatment in HIV infected individuals with active tuberculosis. AB - BACKGROUND & OBJECTIVE: Mycobacterium tuberculosis infection has been shown to result in increased HIV replication and disease progression in HIV-infected individuals through increased immune activation. The objective of this study was to correlate plasma levels of immune activation markers with the presence of tuberculosis (TB) in HIV-infected and uninfected individuals, and to study the changes following anti-tuberculosis treatment. METHODS: Plasma markers of immune activation - neopterin, beta-2-microglobulin (beta2M) and soluble tumour necrosis factor alpha receptor type I (sTNFalpha-RI) were measured by ELISA in 42 HIV positive TB patients (HIV+TB+) undergoing a six-month course of TB chemotherapy. Thirty seven HIV+ persons without active TB, 38 TB patients without HIV infection, and 62 healthy volunteers served as controls. RESULTS: Plasma levels of all three markers were elevated in HIV+ individuals, more so in those with active TB. When HIV+ individuals were further categorized based on CD4+ T cell counts, HIV+TB+ patients with CD4+ T cells counts 1024 microg/ml for the E. coli 27 and 1024 and > 1024 microg/ml for the E. coli ATCC8539, respectively. A synergism between this extract and all aminoglycosides assayed was demonstrated. In the same form synergism between chlorpromazine and kanamycin, amikacin and tobramycin was observed, indicating the involvement of an efflux system. Extracts from H. martiusii could be used as a source of plant derived natural products with modifying antibiotic activity and these products may interact and affect multidrug resistance systems (MDR) as efflux pumps. PMID- 19675387 TI - Hepatoprotective activity of six polyherbal formulations in paracetamol induced liver toxicity in mice. AB - BACKGROUND & OBJECTIVE: Polyherbal formulations available with a wide range of indications like protective to liver, appetite and growth promoters, gastrointestinal and hepatic regulator, as treatment for hepatic dysfunction, for hepatic regeneration as well as liver stimulant and tonic. Despite the widespread use, there is a lack of scientific evidence on their efficacy and safety. This study was undertaken to evaluate the hepatoprotective activity of six commercially available formulations, namely Liv 52, Livergen, Livokin, Octogen, Stimuliv and Tefroliv in acute liver toxicity in mice model induced by paracetamol (PCM). METHODS: Swiss albino mice of either sex were used, divided in 28 groups with six in each group. The dose of the polyherbal formulations was calculated from human dose (20 ml/day) using a standard conversion table. They were given as pretreatment (2.60 ml/kg/day) for 7 days by oral route twice a day prior to PCM administration. Hepatotoxicity was induced by administering a single oral dose of PCM (500 mg/kg bw) on day 8. The study parameters were conducted on day 9. The biochemical parameters included liver enzyme levels alanine tranaminases (ALT), aspartate transaminases (AST) and alkaline phosphatase (ALP). The pharmacological and pathological parameters were phenobarbitone sleeping time and macroscopic and microscopic changes of liver tissues respectively. RESULTS: PCM toxicity significantly increased ALT, AST and ALP (321.00 +/- 87.93, 273.17 +/- 45.68, 257.50 +/- 17.64 IU/l vs normal control, 33.33 +/- 0.61, 89.33 +/- 9.50, 152.17 +/- 11.40 IU/l respectively, P<0.05), prolonged phenobarbitone induced sleeping time (from 277.50 +/- 8.04 min to 335.83 +/- 7.00 min, P<0.05). When PCM higher dose (1g/kg p.o. single dose) was used, the liver tissue, in macroscopic appearance, showed extensive necrosis associated with haemorrhages. Low dose (500 mg/kg p.o. single dose) showed punctate haemorrhagic necrosis of liver tissue. In the microscopic studies, PCM induced toxicity showed haemorrhages, fatty changes and necrosis. The pretreatment in low doses (2.6 ml/kg/day) with liquid formulations of Liv 52 and Livergen reversed the PCM induced liver toxicity. At higher doses (5.2 ml/ kg/day), all the six herbal formulations conclusively showed marked beneficial effects in the studied pharmacological, biochemical and histological parameters. INTERPRETATION & CONCLUSION: The present findings demonstrated the efficacy of polyherbal liquid formulations at two dose levels in PCM induced hepatotoxicity in mice. However, it suggests that a dose adjustment may be necessary to optimize the effects in clinical settings. PMID- 19675388 TI - Influenza a virus induced apoptosis: inhibition of DNA laddering & caspase-3 activity by zinc supplementation in cultured HeLa cells. AB - BACKGROUND & OBJECTIVE: The pathogenesis of influenza virus infection involves virus replication in epithelial cells of the respiratory tract and the consequent degeneration of infected cells. Influenza virus induces cellular degeneration following infection of cultured cells in vitro, and the cytopathic effect (CPE) occurs principally through apoptotic cell death. This study was undertaken to fi nd out the effect of zinc on influenza virus induced apoptosis in cultured HeLa cells. METHODS: The sub-confluent monolayer HeLa cells were used to study the effect of zinc on influenza virus induced apoptosis. The apoptotic markers viz., caspase-3 activity, phagocytic index, morphological changes, and DNA fragmentation were assayed. RESULTS: When HeLa cells were infected with a cell adapted pathogenic strain of influenza A (A/Udorn/ 317/72H(3)N(2)) virus, DNA fragmentation was observed in virus infected cells by 24 h post infection and caspase-3 activity was maximum at 4 h post infection after which it reached to plateau. Treatment of cells with 0.1 5mM concentration of zinc till 8 h post infection inhibited DNA fragmentation and also caspase 3 activity was decreased significantly up to 2 h post infection. INTERPRETATION & CONCLUSION: When the infected HeLa cells were incubated with adherent macrophages, efficient phagocytosis occurred and the release of virus into the culture medium was inhibited. These results suggested that inhibitory effect on influenza virus induced apoptotic death of cultured cells can be determined at an early stage of the infection by treatment of zinc. PMID- 19675389 TI - Prolonged constriction of sciatic nerve affecting oxidative stressors & antioxidant enzymes in rat. AB - BACKGROUND & OBJECTIVE: This study was carried out to determine the effects of reactive oxygen species in the balance between the pro-oxidant and antioxidant levels in experimental peripheral constriction injury induced by silver wire looping of sciatic nerve of rats. METHODS: Rats were divided into experimental group 1 (silver wire ligated) and group 2 (control, sham operated). Functional and behavioural activities were assessed by a modified Basso Beattie Bresnahan (BBB) locomotory rating scale. Mechanical pain intensity was measured with Randall and Selitto apparatus. Foot positioning, toe spread, paw withdrawal threshold and paw withdrawal latency were carried out on days 1, 3, 7, 14, 21 and 28 in rats with chronic pain. Oxidative stress markers such as malondialdehyde (MDA) and advanced oxidation protein products (AOPP) were measured along with antioxidants such as glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH) on day 30 after constriction in sciatic nerve, spinal cord, dorsal root ganglion, dorsal root and ventral root. RESULTS: Significant (P<0.05) increase in MDA, AOPP, SOD and GPx and decrease in the GSH and catalase activities in sciatic nerve, spinal cord, dorsal root ganglion, dorsal root and ventral root were observed in experimental group rats compared to control group. There was no recovery in foot positioning and toe spread. Reduced paw withdrawal threshold and paw withdrawal latency was observed in ligated rats compared to control rats. INTERPRETATION & CONCLUSION: Foot positioning, toe spread, paw withdrawal threshold and paw withdrawal latency with no recovery until day 30 confirmed locomotory deficits, hyperalgesia and neuronal impairment. Oxidative stress evidenced by increased MDA, AOPP and decreased GSH and catalase support the generation of reactive oxygen species in constriction model. The present experimental model for chronic pain induced by silver wire spirally coiled around sciatic nerve may be useful for future studies. PMID- 19675390 TI - Impact of etofenprox (Vectron 20 WP) indoor residual spray on malaria transmission. AB - BACKGROUND & OBJECTIVE: A longitudinal study was carried out to assess the impact of indoor residual spray with Vectron 20 WP (etofenprox) against Anopheles culicifacies and on malaria transmission in a selected village of Dadri primary health centre (PHC), District Gautam Budh Nagar, Uttar Pradesh, India. METHODS: Two villages, namely Patadi and Anandpur in District Gautam Budh Nagar, Uttar Pradesh, with similar malaria incidence and vector prevalence were selected for the present evaluation. In one village two rounds of indoor spraying of etofenprox (0.1 g/m(2)) were done at an interval of 12 wk and the other village was kept as control where no intervention except intensive active surveillance for early detection and prompt treatment (EDPT) was undertaken during the study period. Entomological and epidemiological data were collected using standard procedures. RESULTS: Persistence of the effectiveness of etofenprox against An. culicifacies was observed up to 12 wk. Spraying of etofenprox significantly reduced the density (P<0.001) and proportion of parous An. culicifacies mosquitoes (P<0.05) in the experimental village. There was a significant reduction in malaria cases in the experimental village during the post-spray period when compared to the control village (P<0.05). No adverse effect was reported by the spraymen and inhabitants during and after the spray. INTERPRETATION & CONCLUSION: Indoor residual spray of etofenprox (0.1 g/m(2)) with an interval of three months in between two rounds of spray produced the desired impact in reducing the indoor resting density of vector mosquitoes and also in curtailing malaria transmission in the sprayed village when compared with the control village without spray. PMID- 19675391 TI - Faecal carriage of CTX-M-15-producing Klebsiella pneumoniae in patients with acute gastroenteritis. AB - BACKGROUND & OBJECTIVE: Data on extended-spectrum beta-lactamases (ESBLs) produced by Gram-negative bacteria including Klebsiella pneumoniae especially molecular types of ESBL genes from India are limited. The present study was conducted to investigate the carriage and ESBL contents of multidrug-resistant K. pneumoniae recovered from patients with gastroenteritis in a rural village in southern India. METHODS: Nine K. pneumoniae isolates obtained from 45 stool samples from patients with gastroenteritis from one rural and two urban sites, in southern India were included in the study. Antibiotic susceptibility testing, PCR analysis and sequencing were conducted to characterize the ESBL genes. Clonal relatedness was assessed by pulsed-field gel electrophoresis (PFGE). RESULTS: All the isolates were found to be resistant to at least one of the third generation cephalosporins tested. All the study isolates were confirmed to produce ESBLs. PCR and sequencing revealed the responsible gene to be bla(CTX-M-15). bla(TEM) and bla(SHV) were absent. PFGE indicated that fi ve of seven isolates from villagers were genetically closely related, and in turn were related to isolates from patients in two urban areas in this region. INTERPRETATION & CONCLUSION: Our findings showed that genetically-related isolates of K. pneumoniae producing CTX M-15 were present in multiple areas in southern India. Larger studies need to be done in various geographical regions of the country to better define the molecular epidemiology of ESBL-producing K. pneumoniae and its clinical implications. PMID- 19675392 TI - Effects of progressive muscular relaxation training on quality of life in anxious patients after coronary artery bypass graft surgery. AB - BACKGROUND & OBJECTIVE: Evidences suggest that relaxation therapy may improve psychological outcomes in heart patients. We evaluated the effect of progressive muscular relaxation (PMR) training in decreasing anxiety and improving quality of life among anxious patients after coronary artery bypass graft surgery (CABG). METHODS: This study was an open uncontrolled trial. The sample included 110 anxious patients referred to the cardiac rehabilitation clinic of Tehran Heart Center, Tehran, Iran, during six weeks after coronary artery bypass graft (CABG). Patients were allocated to receive both exercise training and lifestyle education plus relaxation therapy (relaxation group; n=55) or only exercise training beside lifestyle education (control group or the recipient of usual care group; n=55). Duration of the relaxation therapy was 6 wk and in the case of usual care was 8 wk. Both the groups were followed up one month after completion of intervention. Anxiety and quality of life in the two treatment groups were compared. RESULTS: There were no significant differences in overall QOL, state anxiety and trait anxiety scores between the two groups before intervention. Significant reductions in state anxiety (P<0.01) and trait anxiety (P<0.01) levels were observed in relaxation group after intervention compared to control group. Women had high state anxiety and a low quality of life than men in the two groups before intervention. After intervention, there was no difference between men and women in the relaxation group. INTERPRETATION & CONCLUSION: Our findings show that progressive muscular relaxation training may be an effective therapy for improving psychological health and quality of life in anxious heart patients. PMID- 19675393 TI - Plasma paraoxonase activity in patients with systemic sclerosis. AB - BACKGROUND & OBJECTIVE: Systemic sclerosis (SSc) is a connective tissue disease characterized vascular damage and fibrosis. The aim of this study was to investigate the possible relation between systemic sclerosis and paraoxonase which is an antioxidant enzyme on the HDL. METHODS: Twenty nine patients with SSc and 16 healthy subjects (control group) participated in the study. Plasma cholesterol levels, anti-centromere antibody (ACA) levels and paraoxonase (PON) activities were measured. RESULTS: Lower level of high-density lipoprotein (HDL) cholesterol was observed in ACA negative SSc patients than in controls. Paraoxonase activity in ACA positive patients was however found to increase relative to control and ACA negative patient groups. INTERPRETATION & CONCLUSION: Our findings suggested that low HDL level in ACA negative SSc patients might be one of the factors leading to some vascular problems, and increased PON activity in ACA positive SSc group might have some role in the limitation of cutaneous sclerotic process observed in these patients. However, these preliminary findings need to be confirmed with a larger sample. PMID- 19675394 TI - Epidemiology & antibiograms of Vibrio cholerae isolates from a tertiary care hospital in Chandigarh, north India. AB - BACKGROUND & OBJECTIVE: Cholera is endemic in Chandigarh and its surrounding areas. This retrospective study was undertaken over a period of nine years (January 1999-December 2007) from a tertiary care hospital in north India to understand the changing epidemiology aspects and antibiotic resistance patterns in Vibrio cholerae isolates. METHODS: A total of 277 isolates of V. cholerae were included in the study. V. cholerae was identified by standard microbiological procedures. Antibiotic sensitivity testing was performed by disc diffusion method and isolates phage typed. RESULTS: All the isolates were identified as V. cholerae O1 biotype El Tor serotype Ogawa; phage 27 was the predominant type. Men were more commonly affected with maximum number in the age group 0-5 yr. Majority of the isolates were resistant to furazolidone but sensitive to gentamicin and cefotaxime. Resistance pattern to amoxycillin was variable. Three isolates were found to be resistant to ciprofloxacin. All the patients presented during June October coinciding with the monsoon season and a majority were from suburbs. INTERPRETATION & CONCLUSION: The emergence of resistance amongst V. cholerae especially towards ciprofloxacin may significantly influence the control strategies in future outbreaks. Phage 27 remained the predominant type in all the years. Continuous surveillance with regard to drug resistance, early detection and a strong regional commitment may help contain the disease. PMID- 19675395 TI - Challenges of introducing vaginal microbicides in India. PMID- 19675396 TI - South korean study in a public health: preventive medicine and sports environment. AB - OBJECTIVES: The aim of this study was to develop search filters able to retrieve the South Korean scientific output and relating the fields of public health, preventive medicine, and physical education, activity, fitness, exercise and sport in MEDLINE between 2000 and 2007. METHODS: The search filters were constructed and applied in MEDLINE accessed through PubMed according to the affiliation and subject. The language and place of publication were identified and the evaluation of the geographical filter performance was done. RESULTS: The search format was successfully elaborated and applied, and the articles originated, published in Korea and/or abroad focusing on the fields of public health, preventive medicine, physical education, activity, fitness, exercise and sport, added to MEDLINE between 2000 and 2007 were retrieved. Publications in six languages originated in South Korea were detected. CONCLUSIONS: A geographic search filter determined the South Korean research output, and combined with additional filters focused on specific areas. The dynamics of the scientific output followed an increased evolution in all categories. Articles were written mainly in English/Korean. Further research is recommended on developing search strategies in order to retrieve precise and reliable information. PMID- 19675397 TI - Relationship between perceived neighborhood characteristics and vigorous physical activity among adult seoul residents. AB - OBJECTIVES: Vigorous physical activity is a well-known method to promote people s health. This research aims to investigate whether perceived neighborhood characteristics affect vigorous physical activity among adult Seoul residents (aged 19 to 64). METHODS: Utilizing the 2005 Seoul Citizens Health Indicators Survey data, this study estimates the probability of vigorous physical activity. Particular attention is given to the effects of three perceived neighborhood characteristics (satisfaction with relationship to the neighborhood, satisfaction with park and recreational facilities, and satisfaction with public security). Logistic regression models are analyzed separately by gender for the parameter estimation. RESULTS: Vigorous physical activity is positively associated with three perceived neighborhood characteristics for women, while neither significant nor substantive association is found for men. CONCLUSIONS: As vigorous physical activity among Seoul citizens is differentially affected by perceived neighborhood characteristics and by gender, a different approach will be needed to increase vigorous physical activity of men and women in Seoul. PMID- 19675398 TI - Validity of self-reported smoking using urinary cotinine among vocational high school students. AB - OBJECTIVES: This study was conducted to validate self-reported smoking among high school students using urinary cotinine. METHODS: A self report of smoking behavior was collected together with urine sample for cotinine analysis from 130 male and female students in two vocational high school students in November, 2007. Validity and agreement between self-reported smoking and urinary cotinine was analyzed with STATA 9.0 for different definitions of current smokers, and frequent and daily smokers. Urinary cotinine concentration was measured by the DRI Cotinine Assay for urine (Microgenics Corp., Fremont, CA) on Toshiba 200FR. The cut-off point of urinary cotinine was 50 ng/dl. RESULTS: The concentrations of urinary cotinine were significantly different according to the frequency and amount of smoking. Sensitivity and specificity was 90.9% and 91.8% respectively, and the Cohen's kappa value was 0.787 among the current smokers who smoked at least one day during one month preceding the survey. The comparable high sensitivity, specificity, and kappa value were shown also among the other definitions of current smokers, that is, subjective smokers, and weekly smokers. CONCLUSIONS: The results showed the high validity of self-reported smoking among high school students. However, due to the small sample size and limitation of the participants, it is cautious to generalize the results to overall high school students. PMID- 19675399 TI - Determination of appropriate sampling time for job stress assessment: the salivary chromogranin A and cortisol in adult females. AB - OBJECTIVES: This study was conducted to determine the appropriate sampling time of the salivary stress markers, chromogranin A (CgA) and cortisol as objective indices of job stress assessment in adult females. METHODS: The subjects were 20 approximately 39-year-old women (13 office workers, 11 sales-service workers, and 11 college students) who were eligible for the study and free of acute and chronic medical conditions. Salivary CgA and cortisol levels were determined by enzyme-linked immunosorbent assay (ELISA). Saliva samples were collected (2 ml each) at 7:00, 8:00, 10:30, 12:00, 17:30, and 22:30 on a typical day. Salivary CgA and cortisol levels, according to sampling time, were compared among the three groups using general linear model. The full version of the Korean Occupational Stress Scale (KOSS), which includes socioeconomic characteristics, health behavior, work-related characteristics, and BMI, was used to access the subjects' job stress. Multiple regression analysis of the job stressors identified by the KOSS was performed on salivary CgA and cortisol levels. RESULTS: The salivary CgA level peaked at 7:00 (time of awakening), then decreased and were maintained at a low level throughout the day, and increased slightly at 17:30. The salivary cortisol level increased steeply within the 1st hour after awakening, followed by a gradual decrease by 12:00, and was then maintained at a low level throughout the day. The salivary cortisol levels of subjects who worked < or = 5 days per week and graduated from the university were significantly lower at 8:00 (p=0.006). The salivary cortisol levels of non smokers were significantly lower at 7:00 (p=0.040) and 8:00 (p=0.003) compared to smokers. There were no significant differences in salivary CgA and cortisol levels at 10:30 and 12:00 in general characteristics. The regression coefficients on salivary CgA level were significant with interpersonal conflict at 17:30 and job insecurity at 22:30. Regression coefficients on salivary cortisol level were significant with organizational system and total job stressors at 17:30. CONCLUSIONS: We suggest that the appropriate sampling times for the salivary stress markers, CgA and cortisol, are at 7:00 (time of awakening), 8:00 (1 hour after awakening), 17:30 (early evening), and 22:30 (before sleep). PMID- 19675400 TI - [Seroprevalence and risk factors of brucellosis among slaughterhouse workers in Korea]. AB - OBJECTIVES: The incidence of zoonoses in Korea has increased recently. However, the study of high risk groups for zoonoses has not been conducted to date in Korea. Thus, we did this study to obtain data on brucellosis among slaughterhouse workers in Korea. METHODS: We evaluated the structure of slaughterhouses and the process of slaughtering by reviewing the relevant literature and doing field studies. We visited 73 slaughterhouses and 62 residual products handle houses across the country. In addition, we conducted a questionnaire survey of the work activities, and obtained blood samples in order to determine the seroprevalence and risk factors of brucellosis. The titers of brucellosis antibodies were measured using the standard tube agglutination test (SAT). We diagnosed subjects as seropositive for Brucellosis if the titers were more than 1:160. The data collected was evaluated using SPSS ver. 17.0. RESULTS: We included 1,503 subjects and obtained 1,482 blood samples among them: 849 workers involved in slaughtering, 351 handlers of residual products, 190 inspectors and their assistants, and 92 grading testers and their assistants. The seroprevalence of brucellosis among the slaughterhouse workers was 0.8% (95% CI=0.4-1.5). Broken down, the seroprevalence of brucellosis among the workers involved in slaughtering was 0.7% (95% CI=0.3-1.6), the handlers of residual products was 1.7% (95% CI=0.7-3.9) respectively. Risk factors for contracting brucellosis among slaughterhouse workers were being splashed with cattle blood around the mouth, cattle secretions around the body and not putting on protective apron while at work. CONCLUSIONS: An educational program is needed for high risk groups on zoonoses about the prevention of infection. Thus, effective working guidelines for workers who participate in the slaughter of animals must be developed in order to protect them from zoonoses. PMID- 19675401 TI - [Costs of initial cancer care and its affecting factors]. AB - OBJECTIVES: The purposes of this study is to estimate the cost of cancer care after its diagnosis and to identify factors that can influence the cost of cancer care. METHODS: The study subjects were patients with an initial diagnosis one of four selected tumors and had their first two-years of cancer care at a national cancer center. The data were obtained from medical records and patient surveys. We classified cancer care costs into medical and nonmedical costs, and each cost was analyzed for burden type, medical service, and cancer stage according to cancer types. Factors affecting cancer care costs for the initial phase included demographic variables, socioeconomic status and clinical variables. RESULTS: Cancer care costs for the initial year following diagnosis were higher than the costs for the following successive year after diagnosis. Lung cancer (25,648,000 won) had higher costs than the other three cancer types. Of the total costs, patent burden was more than 50% and medical costs accounted for more than 60%. Inpatient costs accounted for more than 60% of the medical costs for stomach and liver cancer in the initial phase. Care for late-stage cancer was more expensive than care for early-stage cancer. Nonmedical costs were estimated to be between 4,500,000 to 6,000,000 won with expenses for the caregiver being the highest. The factors affecting cancer care costs were treatment type and cancer stage. CONCLUSIONS: The cancer care costs after diagnosis are substantial and vary by cancer site, cancer stage and treatment type. It is useful for policy makers and researchers to identify tumor-specific medical and nonmedical costs. The effort to reduce cancer costs and early detection for cancer can reduce the burden to society and improve quality of life for the cancer patients. PMID- 19675402 TI - [Socioeconomic costs of stroke in Korea: estimated from the Korea national health insurance claims database]. AB - OBJECTIVES: To estimate the annual socioeconomic costs of stroke in Korea in 2005 from a societal perspective. METHODS: We identified those 20 years or older who had at least one national health insurance (NHI) claims record with a primary or a secondary diagnosis of stroke (ICD-10 codes: I60-I69, G45) in 2005. Direct medical costs of the stroke were measured from the NHI claims records. Direct non medical costs were estimated as transportation costs incurred when visiting the hospitals. Indirect costs were defined as patients' and caregivers' productivity loss associated with office visits or hospitalization. Also, the costs of productivity loss due to premature death from stroke were calculated. RESULTS: A total of 882,143 stroke patients were identified with prevalence for treatment of stroke at 2.44%. The total cost for the treatment of stroke in the nation was estimated to be 3,737 billion Korean won (KRW) which included direct costs at 1,130 billion KRW and indirect costs at 2,606 billion KRW. The per-capita cost of stroke was 3 million KRW for men and 2 million KRW for women. The total national spending for hemorrhagic and ischemic stroke was 1,323 billion KRW and 1,553 billion KRW, respectively, which together consisted of 77.0% of the total cost for stroke. Costs per patient for hemorrhagic and ischemic stroke were estimated at 6 million KRW and 2 million KRW, respectively. CONCLUSIONS: Stroke is a leading public health problem in Korea in terms of the economic burden. The indirect costs were identified as the largest component of the overall cost. PMID- 19675403 TI - [Apolipoprotein e polymorphism and cognitive function change of the elderly in a rural area, Korea]. AB - OBJECTIVES: The aim of this study is to examine the cognitive function change related to aging, the incidence of cognitive impairment, and the association between apolipoprotein E polymorphism and cognitive impairment through a follow up of the elderly with normal cognitive ability at baseline. METHODS: Two hundred and fifteen subjects aged 65 and over were surveyed in February, 1998 (baseline survey), and their cognitive function was assessed again in 2003 (1st follow-up) and the once again in 2006 (2nd follow-up). Ninety one subjects completed all surveys up through the 2nd follow-up and their cognitive function scores using MMSE-K (Korean Version of the Mini-Mental State Examination) and the distribution of apolipoprotein E allele were analyzed. RESULTS: The cognitive function scores decreased with aging and the difference between baseline and the 2nd follow-up scores of the study increased with the age group. The incidence rate of cognitive impairment through an 8-year follow-up was 38.5% and higher in older age groups. Age was the only significant factor for incidence of cognitive impairment, but there was no significant association between apolipoprotein E genotype and incidence of cognitive impairment. CONCLUSIONS: The cognition of the elderly decreased with aging and the association of apolipoprotein E genotype with incidence of cognitive impairment was not significant in this study. To confirm the association between apolipoprotein E polymorphism and incidence of cognitive impairment further studies will be needed. PMID- 19675404 TI - [A comparative study on comorbidity measurements with Lookback period using health insurance database: focused on patients who underwent percutaneous coronary intervention]. AB - OBJECTIVES: To compare the performance of three comorbidity measurements (Charlson comorbidity index, Elixhauser's comorbidity and comorbidity selection) with the effect of different comorbidity lookback periods when predicting in hospital mortality for patients who underwent percutaneous coronary intervention. METHODS: This was a retrospective study on patients aged 40 years and older who underwent percutaneous coronary intervention. To distinguish comorbidity from complications, the records of diagnosis were drawn from the National Health Insurance Database excluding diagnosis that admitted to the hospital. C-statistic values were used as measures for in comparing the predictability of comorbidity measures with lookback period, and a bootstrapping procedure with 1,000 replications was done to determine approximate 95% confidence interval. RESULTS: Of the 61,815 patients included in this study, the mean age was 63.3 years (standard deviation: +/-10.2) and 64.8% of the population was male. Among them, 1,598 (2.6%) had died in hospital. While the predictive ability of the Elixhauser s comorbidity and comorbidity selection was better than that of the Charlson comorbidity index, there was no significant difference among the three comorbidity measurements. Although the prevalence of comorbidity increased in 3 years of lookback periods, there was no significant improvement compared to 1 year of a lookback period. CONCLUSIONS: In a health outcome study for patients who underwent percutaneous coronary intervention using National Health Insurance Database, the Charlson comorbidity index was easy to apply without significant difference in predictability compared to the other methods. The one year of observation period was adequate to adjust the comorbidity. Further work to select adequate comorbidity measurements and lookback periods on other diseases and procedures are needed. PMID- 19675405 TI - [Endpoint for therapy in allergic diseases: pediatric asthma]. PMID- 19675406 TI - [Therapeutic endpoints in adult asthma]. PMID- 19675407 TI - [One point message of JGL 2007 --proposal of new classification of asthma severity in adult]. PMID- 19675408 TI - [Provocation tests for urticarias including physical urticaria, aspirin urticaria, and food-dependent exercise--induced urticaria]. PMID- 19675409 TI - [Bronchial asthma and genes of the innate immunity]. PMID- 19675410 TI - [Culture of human mast cells from progenitor cells and heterogeneity of human mast cells]. PMID- 19675411 TI - [Single blind food challenge using dried food powder --2nd Report. milk]. AB - AIM: We analyzed data obtained from cow's milk challenge with single-blind manner in our department. SUBJECTS AND METHODS: We have performed 83 cow's milk challenges in the hospital from 1995 to 2005, and patient profiles and results of the challenges were analyzed. RESULTS: Positive rate of cow's milk challenges were 44.6% (37/83), and evoked symptoms by cow's milk challenges were most frequently seen in skin, followed by respiratory tract, and gastrointestinal system. Cases of anaphylaxis were seen in 8.1%. Seventy six percentages of symptoms started within one hour after the challenge, and 56.8% among positive cases were medicated. Adrenaline was injected in 9 cases out of 37 positive challenge cases. And all anaphylaxis patients recovered. We performed 19 open challenges with heated-cow's milk among 37 positive cases, and only one negative case existed. When we compared profiles of negative cow's milk challenges and positive ones, significant higher rate on positive skin prick tests (SPT) against cow's milk and complication of bronchial asthma were recognized in positive group. Sensitivity of SPT was 97.3% and specificity was 17.4%. Sensitivity of IgE CAP-RAST was 100% and specificity was 15.2%. CONCLUSIONS: Single-blinded challenge tests were safe. And they were useful in judgment of the acquisition of oral tolerance in cow's milk allergy. It was only one case that effect of the heating un-matched in food provocation test results. Although we experienced fairly high prevalence of respiratory symptoms during cow's milk challenge tests, appropriate medication especially adrenaline introduction made our test safe enough. PMID- 19675412 TI - [A study of drug aerosols generated by various devices for inhaled steroids]. AB - PURPOSE: To evaluate the performances of various devices for inhaled steroid. METHODS: Aerosol particle diameters were determined using the Aerodynamic Particle Sizer Spectrometer. Conversion ratios to fine particle aerosol were determined using an Andersen non-bubble sampler. RESULTS: The mass median aerodynamic diameter was between 1 and 4 microm for all devices examined, tending to decrease with the increase in a suction flow rate. The percent of drug aerosolized less than 11 microm in diameter at a suction flow rate of 28.3 L/min was 100% for BDP-HFA and CIC-HFA, 87.5% for FP-HFA, 47.0% for BUD-DPI, 6.2% for FP-DPI, and 4.9% for SFC-DPI. The percent of drug aerosolized less than 7.6 microm in diameter at a suction flow rate of 60 L/min was 78.2% for BUD-DPI, 5.9% for FP-DPI, and 6.5% for SFC-DPI. DISCUSSION: For MDI preparations, devices permitting easy synchronization of drug atomization and inspiration exhibited higher inhalation efficiency. For DPI preparations, the BUD-DPI device exhibited higher inhalation efficiency, which was enhanced by increasing the suction flow rate. PMID- 19675419 TI - Discrepancies between clotting and chromogenic assays in congenital coagulation disorders: the hemostatic coin has only one face, chromogenic substrates many. PMID- 19675421 TI - Editorial comment: pancreas. PMID- 19675424 TI - Current Opinion in Hematology. Current world literature. PMID- 19675426 TI - Current world literature. PMID- 19675427 TI - Cardiovascular disease. Preface. PMID- 19675428 TI - (Don't) just start running. PMID- 19675429 TI - Screening for safe sports participation: do for yourself what you tell your patients. PMID- 19675430 TI - Stair use for cardiovascular disease prevention. PMID- 19675431 TI - The challenge for preventive cardiology. PMID- 19675432 TI - What is my risk of developing cardiovascular disease? PMID- 19675433 TI - Risk factor management: a practice guide. PMID- 19675434 TI - Risk factor management: antiatherogenic therapies. AB - Despite the advances in interventional techniques, the management of stable atherosclerosis remains the domain of optimal guideline-oriented therapy. Recent studies on the effects of aggressive lipid lowering on atheroma volume changes using intravascular ultrasound indicate that it is possible to achieve atherosclerosis regression by reaching low-density lipoprotein (LDL) levels less than 75 mg/dl. The pleiotropic anti-inflammatory effects of statins contribute to the reduction of cardiovascular (CV) event observed with aggressive lipid lowering. As a second important strategy to prevent disease progression, lifestyle changes with regular physical exercise are capable of halting the atherosclerotic process and reducing angina symptoms and CV events. Optimal medical therapy, a healthy lifestyle with regular physical exercise, and coronary interventions are not mutually exclusive treatment strategies. Over the last few decades, both have proved to be effective in significantly reducing the CV mortality in the Western world. However, risk factor modification contributed to at least half the effect in the reduction of CV mortality. This figure provides an estimate of what could be achieved if we were to take risk factor modification more seriously - especially in the acute care setting. The knowledge is there: today we have a better understanding on how to stop progression and even induce regression of atherosclerosis. Much research still needs to be done and will be done. In the meantime, however, our primary focus should lie in implementing what is already known. In addition, it is essential not just to treat CV risk factors, but also to treat them to achieve the target values as set by the guidelines of European Society of Cardiology. PMID- 19675435 TI - Practical organisation of preventive cardiology programmes: integrating prevention and rehabilitation. PMID- 19675436 TI - Cardiovascular disease in Europe. PMID- 19675437 TI - Basic and translational research: from molecule, to mouse, to man. PMID- 19675438 TI - Call for action: to promote a heart-healthy environment in Europe. PMID- 19675439 TI - National Coordinators: an active network dedicated to CVD prevention. PMID- 19675440 TI - The political power of heart doctors: with the European Heart Health Charter towards a European policy on cardiovascular disease. PMID- 19675441 TI - Hope for the future: early recognition of increased cardiovascular risk in children and how to deal with it. PMID- 19675442 TI - A modern fairy tale. PMID- 19675443 TI - The cardiovascular prevention and lifestyle quiz for cardiologists. PMID- 19675444 TI - An evidence-based construction of the models of decline of functioning. Part 1: two major models of decline of functioning. AB - The purpose of this study is to confirm the working hypothesis on two major models of functioning decline and two corresponding models of rehabilitation program in an older population through detailed interviews with the persons who have functioning declines and on-the-spot observations of key activities on home visits. A total of 542 community-dwelling older persons (65 years and older) in a suburban city, who were qualified for the services of the Insurance for Long-term Care were visited at their home and they gave two types of interviews: a preliminary semistructured one using an International Classification of Functioning, Disability and Health-based questionnaire and the main detailed one combined with observation of performance of key activities both within and around the home. The results confirmed the two major models of functioning decline and corresponding rehabilitation program: 'stroke model' (acute onset type) was observed in 271 (50.0%) persons and 'disuse syndrome model' (chronic progression type) was observed in 226 (41.4%) persons, along with an additional other model (on account of cognitive problems) in 45 (8.3%) persons. This result confirms that disuse syndrome (deconditioning) should be recognized as an important cause of functioning decline in the older population and a corresponding model of the rehabilitation program should be consciously pursued. This recognition of the importance of disuse syndrome will have a strong impact on clinical thinking about the rehabilitation of the older population. PMID- 19675445 TI - Surveillance, Epidemiology, and End Results (SEER) database analysis of microcystic adnexal carcinoma (sclerosing sweat duct carcinoma) of the skin. AB - BACKGROUND: Microcystic adnexal carcinoma (MAC) is a very rare cancer of the skin. It has only been described previously in case reports and small retrospective series. OBJECTIVE: To analyze and summarize data from the National Cancer Institute, Surveillance, Epidemiology, and End Results (SEER) database regarding MAC. METHODS: The SEER 1973 to 2004 database was investigated, and patients with MAC were identified. A statistical analysis was performed. RESULTS: Two hundred twenty-three patients were identified. Predominant site of disease was the head and neck skin (74%). There was only 1 case of recorded metastatic disease. Lymph nodes were pathologically involved in 1%. The 10-year overall survival was 86.4% (Standard Error [SE]: 3.3%). US census population-matched relative survival was 97.7% at 10 years (SE: 5.2%). LIMITATIONS: This study is limited by the retrospective nature of the SEER database. CONCLUSIONS: MAC is locally invasive, and rarely metastasizes to lymph nodes. Overall and population matched relative survival is excellent. PMID- 19675446 TI - A pilot prospective randomized trial of postoperative epoetin alfa in patients undergoing major operation for upper gastrointestinal malignancy. AB - INTRODUCTION: Strategies to reduce red blood transfusion utilization in cancer patients undergoing operation are needed. HYPOTHESIS: Postoperative epoetin alfa (40,000 units subcutaneous on postoperative days 1 and 7) is associated with improved hematologic parameters in patients undergoing major abdominal surgery for malignancy. MATERIALS AND METHODS: Prospective, blinded, randomized trial of epoetin alfa (40,000 units subcutaneous on postoperative days 1 and 7) versus placebo in patients undergoing major abdominal operation for malignancy. Primary endpoints were immature reticulocyte fraction, reticulocyte count, and hemoglobin, which were measured on postoperative days 4, 7, and between 14 and 20. Secondary endpoints were transfusions and complications in the 2 groups. RESULTS: Forty patients were enrolled. There were no significant differences in immature reticulocyte fraction (P = 0.78), reticulocyte count (P = 0.42), or hemoglobin (0.35) in patients randomized to receive epoetin alfa versus placebo. There was no significant difference in red blood cell transfusion rate or postoperative complications in patients who received epoetin alfa compared with placebo. DISCUSSION: The use of postoperative epoetin alfa (40,000 units subcutaneous on postoperative days 1 and 7) in patients undergoing major operation for abdominal or pelvic malignancy is not supported by this randomized trial. PMID- 19675447 TI - Current treatment strategies for brain metastasis and complications from therapeutic techniques: a review of current literature. AB - Each year approximately 170,000 patients are diagnosed with brain metastasis in the United States, making this the most common intracranial tumor in adults. Historically, treatment strategies focused on the use of whole brain radiation therapy (WBRT) for palliation, yielding a median survival time of only 3 to 6 months. The possible effect of WBRT on cognitive function has generated much concern and debate regarding the use of this modality. Thus, the use of WBRT alone, or in conjunction with other treatment modalities should take into account both risks and benefits, to ensure the best patient outcome with regard to disease state and functional status. The advent of technologies permitting local dose-escalation have clearly increased local control rates, and in select patients, even survival, thereby, further intensifying the debate regarding the use of WBRT. Here, we review the use of WBRT, radiosurgery, and resection for the treatment of brain metastases. Further, we will review the use of radiation sensitizers and blood-brain barrier penetrating cytotoxics such as temozolomide. Finally, we will discuss current treatment strategies for possibly maintaining and improving cognitive function for these patients. PMID- 19675448 TI - Trastuzumab-based therapy for patients with HER2-positive breast cancer: from early scientific development to foundation of care. AB - Human epidermal growth factor receptor 2 (HER2) gene amplification or protein overexpression occurs in 20% to 25% of breast tumors, often leading to an aggressive disease course and poor clinical outcomes. Successful targeting of HER2-positive tumors in preclinical models with trastuzumab has translated to the clinic. In HER2-positive metastatic breast cancer (MBC), trastuzumab provides significant clinical benefit as a monotherapy and in combination with numerous chemotherapies. In the phase III trial of first-line trastuzumab plus chemotherapy, overall response rate (ORR; 50%, P < 0.001), overall survival (25.1 months vs. 20.3 months, P = 0.046) and time to disease progression improved significantly compared with chemotherapy alone (7.4 vs. 4.6 months, P < 0.001), and second-line trastuzumab use after prior trastuzumab has resulted in ORRs of up to 50%. Clinical success in the metastatic setting provided the rationale for assessing trastuzumab in early breast cancer. Four large trials of adjuvant trastuzumab demonstrated significant improvements in disease-free survival (33% 52%) and overall survival (34%-41%) despite tumor size, nodal or hormone-receptor status, and age. New approaches to maximize the clinical benefit of trastuzumab based therapy are under investigation and include novel combinations with other targeted therapies such as bevacizumab, pertuzumab, and lapatinib. PMID- 19675449 TI - Management of metastatic breast cancer: monotherapy options for patients resistant to anthracyclines and taxanes. AB - Resistance to chemotherapeutic agents is a significant obstacle to the effective treatment of metastatic breast cancer (MBC). Anthracycline- and taxane-based regimens are active as first-line treatment for MBC; however, MBC often progresses because of primary or acquired resistance to anthracyclines and taxanes. There are few options for the treatment of patients with anthracycline- and taxane-resistant or taxane-refractory MBC. This article reviews several single agents that have demonstrated activity as treatment for patients with MBC who progress during, or rapidly following, treatment with anthracyclines and taxanes. Results from clinical trials evaluating agents such as ixabepilone, albumin-bound paclitaxel, capecitabine, vinorelbine, pemetrexed, and irinotecan are presented. Single-agent capecitabine is approved for the treatment of patients after failure of anthracyclines and taxanes. Ixabepilone has demonstrated efficacy in patients with MBC resistant to multiple chemotherapeutic agents and is the only agent approved by the Food and Drug Administration as monotherapy for anthracycline-, taxane-, and capecitabine-resistant MBC. Improved treatment strategies and further evaluation of newer agents may reduce the current burden of treatment-resistant or treatment-refractory MBC. PMID- 19675450 TI - Cytokeratin 5 and estrogen receptor immunohistochemistry as a useful adjunct in identifying atypical papillary lesions on breast needle core biopsy. AB - The presence of atypical or usual epithelial proliferations within papillary breast lesions complicates their interpretation on core biopsy. We evaluated the combination of estrogen receptor (ER) and cytokeratin 5 (CK5) as an aid in the distinction of usual duct hyperplasia from atypical proliferations in this setting. Core biopsies from 185 papillary lesions were reviewed and of these, 82 cases were selected for immunohistochemical study based on the presence of an epithelial proliferation between the fibrovascular cores. Fifty-two cases were used as the test set and 30 cases, with subsequent surgical excision, were used as the validation set. The epithelial proliferation was evaluated for staining intensity and percentage of positive cells using CK5 and ER. Expression of both CK5 and ER was significantly different in nonatypical lesions when compared with atypical lesions (P<0.0001). Nonatypical lesions typically showed an ER-low/CK5 high profile and atypical lesions showed an ER-high/CK5-low profile with ER-high expression defined as diffuse strong staining in >90% of cells. CK5-high expression was defined as a mosaic pattern of staining in >20% of cells and CK5 low as absent or staining in <20% of cells. On the basis of their staining profile, 29 of the 30 validation cases were correctly classified using the excision specimen as the gold standard. Patterns and extent of ER and CK5 staining, when used together, are valuable adjunct stains to differentiate usual duct hyperplasia from atypical proliferations within papillary lesions on core biopsy. PMID- 19675451 TI - Myopericytoma in patients with AIDS: a new class of Epstein-Barr virus-associated tumor. AB - Myopericytoma is an uncommon, benign perivascular myoid cell tumor that occurs almost exclusively in somatic soft tissues. We report 2 cases occurring in patients with acquired immunodeficiency syndrome who show unusual clinical and biologic features. One patient presented with a bronchial mass and the other developed mass lesions of the tongue, vocal cord, and brain. Histologically, oval to plump spindly tumor cells with uniform nuclei and scanty cytoplasm formed sheets or cuffs around gaping or narrow vascular spaces. Focally, these areas merged into fascicles of more elongated cells with eosinophilic cytoplasm. The tumor cells were immunoreactive for actin but not desmin, and showed uniform labeling for Epstein-Barr virus (EBV) encoded RNAs on in-situ hybridization. Both patients were alive 5 years after incomplete excision of the lesions. In conjunction with another case reported in the literature, myopericytoma occurring in acquired immunodeficiency syndrome patients exhibits several features distinct from sporadic myopericytoma: presentation in anatomic sites other than somatic soft tissues, frequent presence of multifocal disease, and association with EBV. This tumor type therefore also broadens the spectrum of neoplasms associated with EBV. PMID- 19675452 TI - Plexiform fibromyxoma: a distinctive benign gastric antral neoplasm not to be confused with a myxoid GIST. AB - A great majority of gastric mesenchymal tumors are gastrointestinal stromal tumor (GIST). A rare group of non-GISTs include myxoid mesenchymal neoplasms. In this report, we describe 12 cases of a distinctive gastric tumor, named here as plexiform fibromyxoma. These tumors occurred in 5 men and 7 women of ages 7 to 75 years (median, 41 y). All tumors were located in the gastric antrum and 6 of them also extended into extragastric soft tissues or into the duodenal bulb. The tumors measured from 3 to 15 cm (median, 5.5 cm). Histologically typical was a plexiform intramural growth with multiple micronodules containing paucicellular to moderately cellular myxoid to collagenous and fibromyxoid neoplastic elements. A prominent, sometimes plexiform capillary pattern was typically present. Extramural components included subserosal nodules, and sometimes more cellular, solid nonplexiform spindle cell proliferation. The tumor cells varied from oval to spindled and had limited atypia and mitotic activity < 5/50 high-power fields. Frequent ulceration, mucosal invasion, and vascular invasion (4 cases) had no adverse significance in these tumors. Immunohistochemically, the tumor cells were positive for alpha smooth muscle actin, and variably for CD10, and were consistently negative for KIT, DOG1, CD34, desmin, and S100 protein. No KIT or platelet-derived growth factor receptor alpha mutations were present in the 3 examined cases. None of the 4 patients who were followed from 9 to 20 years (median, 19 y) developed recurrences or metastases. Additional 3 patients survived 14 to 25 years with unknown tumor status. Review of large numbers of mesenchymal tumors in the esophagus and intestines did not reveal similar tumors. Plexiform fibromyxoma is a distinctive benign gastric antral neoplasm that should be separated from GIST, nerve sheath tumors, and other fibromyxoid neoplasms. PMID- 19675453 TI - Invasive tubular carcinoma of the breast frequently is clonally related to flat epithelial atypia and low-grade ductal carcinoma in situ. AB - Low-grade precursor lesions, such flat epithelial atypia (FEA), low-grade ductal carcinoma in situ (lg-DCIS), and lobular neoplasia (LN) often coexist with invasive tubular carcinomas (TCs) of the breast. To evaluate a possible clonal relationship, we have examined a series of 27 TC and the surrounding putative precursor lesions using loss of heterozygosity analysis and mitochondrial DNA sequencing. In these lesions (22 FEA, 10 lg-DCIS, 3 LN), loss of heterozygosity was most frequently observed on the long arm of chromosome 16 as well as at chromosome 8p21, 3p14, 1p36 and 11q14 with a high degree of homology of allelic losses between FEA, lg-DCIS and tubular carcinomas. In the adjacent invasive tubular carcinomas, mitochondrial DNA sequencing revealed identical mutation patterns in 50% of the lg-DCIS and in 12 of 21 (57%) informative cases of FEA. No direct association was seen between TC and LN or columnar cell lesions without nuclear atypia. Our data indicate, that in the majority of cases lg-DCIS and FEA are directly related to tubular breast cancer with a possible precursor role. PMID- 19675454 TI - Primary bone diffuse large B-cell lymphoma: clinicopathologic study of 21 cases and review of literature. AB - Primary bone diffuse large B-cell lymphomas (PB-DLBCL) are uncommon extranodal lymphomas. Herein, we report the clinical, pathologic, immunohistochemical, and molecular features of 21 cases of PB-DLBCL. The mean age of the patients was 54 years (range: 13 to 85 y). The male and female ratio was 1.6:1. The tumors consisted of diffuse sheets of large atypical cells or a polymorphous mixture of small-to-large cells with large multilobated nuclei, fine chromatin, and inconspicuous to prominent nucleoli. Twelve (57%) cases were non-germinal center B (GCB) and 9 (43%) were GCB subtype based on immunohistochemical classification. B-cell lymphomas (BCL)-2 was positive in 17/21 (81%), TP53 in 11/21 (52%) positive and the mean MIB-1 index was 57%. Polymerase chain reaction showed 10 cases with immunoglobulin heavy-chain (IGH) and 4 cases with IGH/BCL-2 gene rearrangement. The fluorescence in-situ hybridization analyses showed 14% of cases with BCL-6, 19% of cases with BCL-2, and 9% of cases with C-MYC gene rearrangement. Age <60 years and complete response to initial treatment were significant predictors of survival outcome (P< or =0.05). Even though no association was observed between the subtype of PB-DLBCL (GCB vs. non-GCB), BCL2, TP53, MIB1 index and overall survival (P>0.05), due to small sample size, and variability in treatment received, this analysis may be interpreted with caution. PMID- 19675455 TI - The effects of chronic cocaine exposure on impulsivity in rats. AB - Chronic exposure to cocaine increases impulsive behavior, leading to a reduced preference for a larger, delayed reinforcer over a smaller, immediate reinforcer. This study examined the development of impulsivity over multiple days of cocaine exposure and cessation of cocaine. Male Sprague-Dawley rats were trained on a discrete-trials delay-discounting task, during which they chose between a small, immediate reinforcer of one food pellet and a large reinforcer of 3 food pellets after an adjusted delay (0, 10, 20, 40, and 60 s). When stable preferences were established, rats received daily injections of deionized water or cocaine (3, 7.5, 15 mg/kg) 5 min before the delay-discounting task for 9 days. All groups showed an increased preference for the smaller reinforcer as delay to the larger reinforcer increased. Repeated exposure to 7.5 or 15 mg/kg cocaine further decreased preference for the larger reinforcer over the 9 days. When cocaine administration was discontinued, preference for the larger reinforcer returned to baseline levels in the 7.5 mg/kg group, but remained depressed in the 15 mg/kg group. These findings indicate that continuing exposure to cocaine dose dependently decreases choice for the large reinforcer over time, and that the bias remains when cocaine is no longer administered, and that the recovery after high doses of cocaine occurs slowly. PMID- 19675458 TI - Current world literature. Renal and urological problems. End-of-life management. Bone and haematological problems. PMID- 19675459 TI - Ethical guidelines in pandemic influenza: recommendations of the Ethics Subcommittee of the Advisory Committee of the Director, Centers for Disease Control and Prevention. AB - Because of the importance of including ethical considerations in planning efforts for pandemic influenza, in February 2005 the Centers for Disease Control and Prevention requested that the Ethics Subcommittee of the Advisory Committee to the Director develop guidance that would serve as a foundation for decision making in preparing for and responding to pandemic influenza. Specifically, the ethics subcommittee was asked to make recommendations regarding ethical considerations relevant to decision making about vaccine and antiviral drug distribution prioritization and development of interventions that would limit individual freedom and create social distancing. The ethics subcommittee identified a number of general ethical considerations including identification of clear goals for pandemic planning, responsibility to maximize preparedness, transparency and public engagement, sound science, commitment to the global community, balancing individual liberty and community interests, diversity in ethical decision making, and commitment to justice. These general ethical considerations are applied to the issues of vaccine and antiviral drug distribution and use of community mitigation interventions. PMID- 19675460 TI - Auditory N1 component to gaps in continuous narrowband noises. AB - OBJECTIVES: To determine whether (1) the auditory N1 component can be elicited to gaps in continuous narrowband noises, (2) psychophysical and electrophysiological gap thresholds (PGTs and EGTs) are similar to one another, and (3) EGTs are the same for all narrowband noise center frequencies. DESIGN: PGTs and EGTs were obtained from 18 normal-hearing young-adult listeners to gaps in continuous narrowband noises with center frequencies of 0.5, 1, or 4 kHz. PGTs were obtained with a modified Bekesy-type tracking paradigm, whereas EGTs were obtained to 2-, 5-, 10-, 20-, or 50-msec gaps presented every 2.2 sec. RESULTS: (1) The auditory N1 component was recorded to gaps in narrowband noises, although they appeared morphologically different from cortical potentials obtained using the continuous broadband noise. (2) At center frequencies of 1 and 4 kHz, psychometric functions revealed close similarity between PGTs and EGTs. However, different results were present for the 0.5-kHz narrowband noise, attributed to stimulus artifact. (3) EGTs were approximately 10 msec for most participants at 1 and 4 kHz, but 20 msec at 0.5 kHz, corroborating other studies showing increases in gap threshold with lower center frequencies. CONCLUSIONS: The auditory N1 component can be recorded to gaps in continuous narrowband noises whose gap thresholds are grossly similar to those obtained psychophysically. The differences found between PGTs and EGTs with different narrowband noise center frequencies call for further investigation of narrowband noise stimuli for the study of temporal resolution. PMID- 19675456 TI - Anxiolytic-like effects of the neurokinin 1 receptor antagonist GR-205171 in the elevated plus maze and contextual fear-potentiated startle model of anxiety in gerbils. AB - Gerbils show a neurokinin (NK)1 receptor pharmacological profile, which is similar to that observed in humans, and thus have become a commonly used species to test efficacy of NK1 receptor antagonists. The aim of this study was to determine whether systemic administration of the NK1 receptor antagonist GR 205171 produced anxiolytic-like effects in the elevated plus maze and in a novel contextual conditioned fear test using fear-potentiated startle (FPS). On the elevated plus maze, treatment with GR-205171 at 0, 0.3, 1.0, and 5.0 mg/kg doses, 30 min before testing produced anxiolytic-like effects in an increasing dose response manner as measured by the percentage of open arm time and percentage of open arm entries. For contextual fear conditioning, gerbils were given 10 unsignaled footshocks (0.6 mA) at a 2-min variable interstimulus interval in a distinctive training context. Twenty-four hours after training, gerbils received treatment of GR-205171 at 0, 0.3, 1.0, and 5.0 mg/kg doses, 30 min before testing in which startle was elicited in the same context in which they were trained. Contextual FPS was defined as an increase in startle over pretraining baseline values. All drug dose levels (0.3, 1.0, and 5.0 mg/kg) significantly attenuated contextual FPS when compared with the vehicle control group. A control group, which received testing in a different context, showed little FPS. These findings support other evidence for anxiolytic activity of NK1 receptor antagonists and provide a novel conditioned fear test that may be an appropriate procedure to test other NK1 antagonists for preclinical anxiolytic activity in gerbils. PMID- 19675461 TI - Trends in referral to outpatient cardiac rehabilitation in the Hunter Region of Australia, 2002-2007. AB - BACKGROUND: Cardiac rehabilitation (CR) is an underutilized evidence-based treatment. We described trends in referral to outpatient CR (OCR) and the factors associated with referral. DESIGN: Cross-sectional survey data provided by Hunter residents aged 20 years or older discharged from public hospitals in the region between 2002 and 2007 with an OCR eligible diagnosis were extracted from the Hunter New England Heart and Stroke Register database. METHODS: Trends in referral were determined using the chi test for trend. Factors associated with referral were examined using multiple logistic regression. RESULTS: Sixty-five percent (4971 of 7678) of patients provided sufficient data for inclusion in the analysis. Approximately half of the patients reported being referred to OCR. No increase over time was observed. Factors associated with referral were age less than 70 years, male sex, being married, urban residence, at least one admission to the tertiary referral hospital for cardiology, at least one admission for acute myocardial infarction, revascularization, no admissions for congestive heart failure, a self-reported history of high cholesterol, and no history of stroke or atrial fibrillation. CONCLUSION: Access to this treatment of proven benefit remained suboptimal despite the provision of new programs and expansion of existing programs. Automatic referral, which is recommended in Australia, should be standard practice. PMID- 19675462 TI - Early symptomatic presbyopes--what correction modality works best? AB - PURPOSE: To compare the performance of a low-addition silicone hydrogel multifocal soft lens with other soft lens correction options in a group of habitual soft lens wearers of distance correction who are symptomatic of early presbyopia. METHOD: This clinical study was designed as a prospective, double masked, randomized, crossover, dispensing trial consisting of four 1-week phases, one for each of the correction modalities: a low-addition silicone hydrogel multifocal soft lens, monovision, habitual correction, and optimized distance visual correction. The prescriptions of all modalities were finalized at a single fitting visit, and the lenses were worn according to a randomized schedule. All lenses were made from lotrafilcon B material. A series of objective vision tests were conducted: high- and low-contrast LogMAR under high- and low-room lighting conditions, stereopsis, and critical print size. A number of other data collection methods used were novel: some data were collected under controlled laboratory-based conditions and others under "real-world" conditions, some of which were completed on a BlackBerry hand-held communication device. RESULTS: All participants were able to be fit with all four correction modalities. Objective vision tests showed no statistical difference between the lens modalities except in the case of low-contrast near LogMAR acuity under low-lighting levels where monovision (+0.29 +/- 0.10) performed better than the multifocal (+0.33 +/- 0.11, P=0.027) and the habitual (+0.37 +/- 0.12, P<0.001) modalities. Subjective ratings indicated a statistically better performance provided by the multifocal correction compared with monovision, particularly for the vision associated with driving tasks such as driving during the daytime (93.3 +/- 8.8 vs. 84.2 +/- 23.7, P=0.05), at nighttime (88.8 +/- 11.7 vs. 74.9 +/- 23.6, P=0.001), any associated haloes or glare (92.0 +/- 10.6 vs. 78.0 +/- 22.8, P=0.003), and observing road signs (90.1 +/- 11.8 vs. 79.4 +/- 20.2, P=0.027). Preference for the multifocal compared with monovision was also reported when watching television (95.0 +/- 6.4 vs. 82.6 +/- 20.1, P=0.001) and when changing focus from distance to near (87.0 +/- 13.4 vs. 66.1 +/- 32.2, P<0.001). CONCLUSIONS: For this group of early presbyopes, the AIR OPTIX AQUA MULTIFOCAL--Low Add provided a successful option for visual correction, which was supported by the results of subjective ratings, many of which were made during or immediately after performing such activities as reading, using a computer, watching television, and driving. These results suggest that making a prediction of "success or not" based on consulting room acuity tests alone is probably unwise. PMID- 19675463 TI - The association between food insecurity and mortality among HIV-infected individuals on HAART. AB - BACKGROUND: Food insecurity is increasingly recognized as a barrier to optimal treatment outcomes, but there is little data on this issue. We assessed associations between food insecurity and mortality among HIV-infected antiretroviral therapy-treated individuals in Vancouver, British Columbia, and whether body max index (BMI) modified associations. METHODS: Individuals were recruited from the British Columbia HIV/AIDS drug treatment program in 1998 and 1999 and were followed until June 2007 for outcomes. Food insecurity was measured with the Radimer/Cornell questionnaire. Cox proportional hazard models were used to determine associations between food insecurity, BMI, and nonaccidental deaths when controlling for confounders. RESULTS: Among 1119 participants, 536 (48%) were categorized as food insecure and 160 (14%) were categorized as underweight (BMI < 18.5). After a median follow-up time of 8.2 years, 153 individuals (14%) had died from nonaccidental deaths. After controlling for adherence, CD4 counts, and socioeconomic variables, people who were food insecure and underweight were nearly 2 times more likely to die (adjusted hazard ratio = 1.94, 95% confidence interval = 1.10 to 3.40) compared with people who were not food insecure or underweight. There was also a trend toward increased risk of mortality among people who were food insecure and not underweight (adjusted hazard ratio = 1.40, 95% confidence interval = 0.91 to 2.05). In contrast, people who were underweight but food secure were not more likely to die. CONCLUSIONS: Food insecurity is a risk factor for mortality among antiretroviral therapy-treated individuals in British Columbia, particularly among individuals who are underweight. Innovative approaches to address food insecurity should be incorporated into HIV treatment programs. PMID- 19675464 TI - Differential survival benefit of universal HAART access in Brazil: a nation-wide comparison of injecting drug users versus men who have sex with men. AB - OBJECTIVE: Brazil accounts for approximately 70% of injection drug users (IDUs) receiving highly active antiretroviral therapy (HAART) in low-income/middle income countries. We evaluated the impact of HAART availability/access on AIDS related mortality among IDUs versus men who have sex with men (MSM). DESIGN: Nation-wide analysis on Brazilian IDU and MSM diagnosed with AIDS in 2000-2006. METHODS: Four national information systems were linked, and Cox regression was used to assess impact of HAART availability/access on differential AIDS-related mortality. RESULTS: Among 28,426 patients, 6777 died during 87,792 person-years of follow-up. Compared with MSM, IDU were significantly less likely to be receiving HAART, to have ever had determinations for CD4 or viral load. After controlling for confounders, IDU had a significantly higher risk of death (adjusted hazard ratio: 1.94; 95% confidence interval: 1.84 to 2.05). Among the subset that had at least 1 CD4 and viral load determination, higher risk of death among IDU persisted (hazard ratio: 1.82; 95% confidence interval: 1.58 to 2.11). Nonwhite ethnicity significantly increased this risk, whereas prompt HAART uptake after AIDS diagnosis reduced the risk of death. After controlling for spatially correlated survival data, AIDS-related mortality remained higher in IDU than in MSM. CONCLUSIONS: Despite free/universal HAART access, differential AIDS-related mortality exists in Brazil. Efforts are needed to identify and eliminate these health disparities. PMID- 19675465 TI - Characteristics of HIV voluntary counseling and testing clients before and during care and treatment scale-up in Moshi, Tanzania. AB - OBJECTIVES: We evaluated changes in characteristics of clients presenting for voluntary counseling and testing (VCT) before and during care and treatment center (CTC) scale-up activities in Moshi, Tanzania, between November 2003 and December 2007. METHODS: Consecutive clients were surveyed after pretest counseling, and rapid HIV antibody testing was performed. Trend tests were used to assess changes in seroprevalence and client characteristics over time. Multivariable logistic regression models were used to estimate the contribution of changes in sociodemographic and behavioral risk characteristics, and symptoms, to changes in seroprevalence before and during CTC scale-up. RESULTS: Data from 4391 first-time VCT clients were analyzed. HIV seroprevalence decreased from 26.2% to 18.9% after the availability of free antiretroviral therapy and expansion of CTCs beyond regional and referral hospitals. Seroprevalence decreased by 27 % for females (P = 0.0002) and 34% for males (P = 0.0125). Declines in seropositivity coincided with decreases in symptoms among males and females (P < 0.0001) and a more favorable distribution of sociodemographic risks among females (P = 0.002). No changes in behavioral risk characteristics were observed. CONCLUSIONS: Concurrent with the scale-up of CTCs, HIV seroprevalence and rates of symptoms declined sharply at an established freestanding VCT site in Moshi, Tanzania. If more HIV-infected persons access VCT at sites where antiretrovirals are offered, freestanding VCT sites may become a less cost effective means for HIV case finding. PMID- 19675466 TI - Apnea training effects on swimming coordination. AB - Triathletes and elite breath-hold divers show an adaptive response to hypoxia induced by repeated epochs of breath holding. We hypothesized that hypoxic training could also improve swimming coordination. Before and after a 3-month breath-hold training program, 4 male swimmers performed a maximal incremental test on bicycle and a 50-m front crawl race at maximal speed without breathing so that interarm coordination could be assessed. Swim velocity, stroke rate (SR), stroke length (SL), and the arm stroke phases were calculated from video analysis. Arm coordination was quantified in terms of an index of coordination (IdC) based on the time gap between the propulsive phases of each arm. After apnea training, the forced expiratory volume in 1 second was higher (4.85 +/- 0.78 vs. 4.94 +/- 0.81 L, p < 0.05), with concomitant increases in VO2peak, minimal arterial oxygen saturation, and respiratory compensation point values (W and W x kg(-1)) during the incremental test. Swimming performance was not improved (clean velocity and time on 50 m); however, SR was decreased and SL and IdC were increased. These results indicate that apnea training improves effectiveness at both peak exercise and submaximal exercise and can also improve swimming technique by promoting greater propulsive continuity. PMID- 19675467 TI - Correcting the use of the term "power" in the strength and conditioning literature. AB - Many strength and conditioning papers have incorrectly adopted the colloquial use of the term "power" as a measure of short-term, high-intensity muscular performance despite a long history of research and editorials critical of this practice. This has lead to confusion, incorrect interpretations, and conflicting results in the literature. This paper summarizes the scientific evidence on external mechanical power as a short-term, high-intensity neuromuscular (anaerobic) performance or training variable. Many problems in the measurement and use of power in strength and conditioning research were identified, as well as problems in the use of the vertical jump as a field test of power. A critical review of the biomechanics, measurement, and training research does not support this colloquial use of the term "power." More research is needed that improves our understanding of the domains of muscular strength or neuromuscular performance, as well as partial correlation and multiple regression analyses to document the unique associations between these domains, biomechanical variables, training effects, and sport performance. Strength and conditioning research should limit the use of the term power to the true mechanical definition and provide several specific and measurement details on this measurement. PMID- 19675468 TI - Strength and power determinants of grinding performance in America's Cup sailors. AB - The purpose grinding is a physically demanding component of America's Cup sailing that is important to overall team performance, but little research is available on the determinants of grinding performance. We examined the relationship between various measures of muscular performance and the performance of upper-body grinding. Eleven elite male America's Cup sailors (33.9 +/- 5.5 yr, 97.8 +/- 12.5 kg, 186.0 +/- 7.1 cm) who performed grinding as part of their on-board role with extensive strength training experience participated in this study. Muscular performance testing examined the force, velocity, and power capabilities of the upper-body musculature, with upper-body push (bench press) and pull (bench pull) movements performed across loads of 10-100% of 1 repetition maximum (1RM). Functional grinding performance was examined for both forward and backward grinding and at 2 different resistances (moderate = 48 N x m, heavy = 68 N x m) using a land-based ergometer. Bench press 1RM and maximum force capability were the measures demonstrating the strongest correlation with forward grinding performance (r = 0.88-0.99 and 0.87-0.99, respectively), with the relationship increasing with grinding load. For backward grinding, there was a very strong relationship with bench pull maximum power (r = 0.85-0.98) in addition to 1RM (r = 0.90-0.95) and maximum force (r = 0.87-0.95). It appears that although maximal strength is a crucial muscular performance characteristic for grinding performance in all conditions, for backward grinding, there is the additional need to focus on the development of speed strength/power to maximize performance gains. This information was used by the Emirates Team New Zealand physical conditioner to develop a conditioning intervention to help improve grinding performance. PMID- 19675469 TI - Precompetition warm-up in elite and subelite rhythmic gymnastics. AB - The aim of this study was to investigate which precompetition warm-up methodologies resulted in the best overall performance in rhythmic gymnastics. The coaches of national and international clubs (60 elite and 90 subelite) were interviewed. The relationship between sport performance and precompetition warm up routines was examined. A total of 49% of the coaches interviewed spent more than 1 hour to prepare their athletes for the competition, including 45 minutes dedicated to warm-up exercises. In spite of previous studies' suggestions, the time between the end of warm-up and the beginning of competition was more than 5 minutes for 68% of those interviewed. A slow run was the activity of choice used to begin the warm-up (96%). Significant differences between elite and subelite gymnasts were found concerning the total duration of warm-up, duration of slow running, utilization of rhythmic steps and leaps during the warm-up, the use of dynamic flexibility exercises, competition performances repetition (p < 0.01), and utilization of imagery (p < 0.05). A precompetition warm-up in rhythmic gymnastics would include static stretching exercises at least 60 minutes prior to the competition starting time and the active stretching exercises alternated with analytic muscle strengthening aimed at increasing muscle temperature. Rhythmic gymnastics coaches at all levels can use this data as a review of precompetition warm-up practices and a possible source of new ideas. PMID- 19675470 TI - Lower-extremity ground reaction forces in youth windmill softball pitchers. AB - Ground reaction forces are important in pitching given that the only external contact a pitcher has is between the foot and the ground. Windmill softball pitchers are routinely seen clinically for injuries to the lower extremities, and lower-extremity kinetics have not been well studied. The purpose of this study was to investigate the relationships between ground reaction forces and throwing mechanics in youth windmill pitchers and to provide a scientific basis for the improvement of preventive and rehabilitative protocols. Fifty-three youth softball pitchers were tested in an indoor facility. High-speed video and force plate data were collected for fastballs from each pitcher. Average ball speed was 25 m/sec. Peak vertical ground reaction force averaged 139 % body weight (BW), peak anterior force averaged 24 %BW, and the medially directed component of the ground reaction force averaged 42 %BW. Loading rates to peak force in all 3 directions were high. Preventive and rehabilitative protocols for windmill softball pitchers can begin to be improved on the basis of knowledge of the magnitudes and times to peak forces under the stride foot. PMID- 19675471 TI - Physical determinants of tennis performance in competitive teenage players. AB - It is unclear how physical attributes influence tennis-specific performance in teenage players. The aims of this study were (a) to examine the relationships between speed, explosive power, leg stiffness, and muscular strength of upper and lower limbs; and (b) to determine to what extent these physical qualities relate to tournament play performance in a group of competitive teenage tennis players. A total of 12 male players aged 13.6 +/- 1.4 years performed a series of physical tests: a 5-m, 10-m, and 20-m sprint; squat jump (SJ); countermovement jump (CMJ); drop jump (DJ); multi-rebound jumps; maximum voluntary contraction of isometric grip strength; and plantar flexor of the dominant and nondominant side. Speed (r = 0.69, 0.63, and 0.74 for 5-, 10-, and 20-m sprints, respectively), vertical power abilities (r = -0.71, -0.80 and -0.66 for SJ, CMJ, and DJ, respectively), and maximal strength in the dominant side (r = -0.67 and -0.73 for handgrip and plantar flexor, respectively) were significantly correlated with tennis performance. However, strength in the nondominant side (r = -0.29 and -0.42 for handgrip and plantar flexor) and leg stiffness (r = -0.15) were not correlated with the performance ranking of the players. It seems that physical attributes have a strong influence on tennis performance in this age group and that an important asymmetry is already observed. By monitoring regularly such physical abilities during puberty, the conditioning coach can modify a program to compensate for the imbalances. This would in turn minimize the risks of injuries during this critical period. PMID- 19675472 TI - An analysis of playing positions in elite men's volleyball: considerations for competition demands and physiologic characteristics. AB - The purpose of this study was to investigate the physiologic demands, physiologic characteristics, and jumping ability of different playing positions in elite male volleyball players. The first investigation involved an analysis of 16 international men's volleyball matches. The second investigation involved an analysis of the anthropometric and jump performance characteristics of 142 Development National Team (DNT) and Senior National Team (SNT) international volleyball players. Mean (+/-SD) frequency of block jumps for Middles (11.00 +/- 3.14) was significantly greater than for Setters (6.25 +/- 2.87, p < 0.001) and Outsides (6.50 +/- 3.16, p < 0.001). Attack jumps were performed more frequently by Middles (7.75 +/- 1.88), and this was found to be significantly more than for Setters (0.38 +/- 1.06, p < 0.001) and Outsides (5.75 +/- 3.25, p < 0.01). Middles were taller than Outsides and Setters (p < 0.001). Consequently, Middles had a significantly higher reach and greater body mass than Outsides (p < 0.001, p < 0.003) and Setters (p < 0.001, p < 0.001). Both Middles and Outsides had superior countermovement vertical jump (CMVJ) and spike jump (SPJ) scores compared with Setters (p < 0.001). Position-specific comparisons between DNT players and SNT players demonstrated that the SNT players were superior in relative CMVJ and SPJ scores (p < 0.05), with a large magnitude of effect (d > 0.99). The results of this study highlight the large jumping and landing demands placed on the taller and heavier players in the middle position. In addition to establishing the magnitude of difference in jumping ability between junior and senior national team players, the results also provide a comprehensive data set that may assist with talent identification and talent development for aspiring male volleyball players. PMID- 19675473 TI - Anthropometric and metabolic determinants of 6,000-m rowing ergometer performance in internationally competitive rowers. AB - The aim of this study was to examine the anthropometric and metabolic determinants of performance during 6,000-m of rowing on an ergometer. The sample comprised 25 internationally successful male heavyweight rowers (mean +/- SD: age 22.2 +/- 4.8 years, rowing experience 8.8 +/- 4.6 years, stature 1.91 +/- 0.05 m, body mass 91.7 +/- 5.9 kg, maximal oxygen uptake 5.53 +/- 0.30 L x min(-1)). The rowers completed an incremental maximal exercise test on a rowing ergometer and, within 2 weeks of this test, also completed a 6,000-m rowing ergometer time trial (mean +/- SD: 1195.4 +/- 36.1 seconds). The strongest correlates (r > 0.5, p < 0.05) with performance were lean body mass (r = -0.767), power output at ventilatory threshold (r = -0.743), power output at maximal oxygen uptake (r = 0.732), body mass (r = -0.693), chest girth (r = -0.598), relaxed arm girth (r = 0.574), forced vital capacity (r = -0.519), and arm span (r = -0.505). Stepwise multiple linear regression procedures indicated that the model comprising a combination of anthropometric and metabolic variables is the best predictor of performance (adjusted R2 = 0.722), followed by models comprising anthropometric (adjusted R2 = 0.575) and metabolic (adjusted R2 = 0.530) variables alone. The results suggest that 6,000-m ergometer performance is determined mainly by power output at ventilatory threshold (58.7% of explained variance). Based on the obtained correlations and regression models, it can be concluded that rowers competing over a 6,000 m distance on a rowing ergometer should devote their training time to the improvement of lean body mass and to the improvement of power output corresponding to ventilatory threshold. PMID- 19675474 TI - Are anthropometric, flexibility, muscular strength, and endurance variables related to clubhead velocity in low- and high-handicap golfers? AB - The present study assessed the anthropometric profile (International Society for the Advancement of Kinanthropometry protocol), flexibility, muscular strength, and endurance of 20 male golfers. These data were collected in order to determine: a) the relationship between these kinanthropometric measures and clubhead velocity; and b) if these measures could distinguish low-handicap (LHG) and high-handicap (HHG) golfers. Ten LHG (handicap of 0.3 +/- 0.5) and 10 HHG (handicap of 20.3 +/- 2.4) performed 10 swings for maximum velocity and accuracy with their own 5-iron golf club at a wall-mounted target. LHG hit the target significantly more (115%) and had a 12% faster clubhead velocity than HHG (p < 0.01). The LHG also had significantly (28%) greater golf swing-specific cable woodchop (GSCWC) strength (p < 0.01) and tendencies for greater (30%) bench press strength and longer (5%) upper am and total arm (4%) length and less (24%) right hip internal rotation than HHG (0.01 < p < 0.05). GSCWC strength was significantly correlated to clubhead velocity (p < 0.01), with bench press and hack squat strength as well as upper arm and total arm length also approaching significance (0.01 < p < 0.05). Golfers with high GSCWC strength and perhaps greater bench press strength and longer arms may therefore be at a competitive advantage, as these characteristics allow the production of greater clubhead velocity and resulting ball displacement. Such results have implications for golf talent identification programs and for the prescription and monitoring of golf conditioning programs. While golf conditioning programs may have many aims, specific trunk rotation exercises need to be included if increased clubhead velocity is the goal. Muscular hypertrophy development may not need to be emphasized as it could reduce golf performance by limiting range of motion and/or increasing moment of inertia. PMID- 19675475 TI - Using near-infrared spectroscopy to determine maximal steady state exercise intensity. AB - Maximal steady state (MSS) speed can be determined from blood lactate concentration (HLa); however, this method is not optimal. The purpose of this study was to determine whether near-infrared spectroscopy (NIRS) technology could be used to detect a breakpoint in percent oxygen saturation (StO2) of the muscle and whether the determined breakpoint exercise intensity could be used to determine MSS exercise intensity. Sixteen distance runners and triathletes (men = 9, VO2max = 64.9 +/- 4.9 ml x kg(-1) x min(-1), women = 7, VO2max = 50.8 +/- 7.0 ml x kg(-1) x min(-1)) completed an incremental exercise test. A change from linearity when plotting StO2 or HLa vs. running speed was defined as the breakpoint. The subjects then completed constant speed runs to determine maximal lactate steady state (MLSS). In 12 subjects, breakpoints were identified for both HLa and StO2 values. Predicted MLSS velocities from HLa breakpoint (12.76 +/- 1.63 km x h(-1)), StO2 breakpoint (12.84 +/- 1.58 km x h(-1)), and 4 mM HLa (13.49 +/- 1.71 km x h(-1)) methods from the incremental test did not differ from MLSS speeds (13.04 +/- 2.03 km x h(-1)). A Bland and Altman analysis of agreement between the MLSS and the StO2 breakpoint speeds resulted in a mean difference of 0.14 +/- 0.36, whereas the mean difference between MLSS and HLa breakpoint speeds was 0.19 +/- 0.43. During the incremental test, no StO2 breakpoint was determined in 2 subjects, whereas 2 subjects had no HLa breakpoint. The results of this study lead us to conclude that the NIRS determination of StO2 is a noninvasive technique that is comparable with HLa in determining MSS intensity and therefore appropriate for use in determining exercise training intensity. PMID- 19675476 TI - Use of aggregate fitness indicators to predict transition into the National Hockey League. AB - Athletes (n = 345) invited to the annual combine conducted by the National Hockey League (NHL) prior to the entry draft were administered tests to measure upper body strength, lower body power, aerobic and anaerobic energy systems, and body composition. Their common variance was extracted using factor analysis from which an overall composite index was derived. A score on this index in the 90th percentile is associated with 72% and 60% probability of playing in the NHL within 4 years after the draft for defensemen and forwards, respectively. These findings demonstrate that by taking into account the shared variance on standard tests of fitness, it is possible to use the athlete's results to gauge his potential for playing in the NHL. PMID- 19675477 TI - Influence of hydration status on pacing during trail running in the heat. AB - The purpose of this study was to determine the influence of hydration status on pacing of trail runners in the heat (wet bulb globe temperature = 26.2 +/- 1.8 degrees C). A randomized, crossover design was used and the participation occurred within a 2-week period. Seventeen competitive, well-trained distance runners (9 men, 8 women, age 27 +/- 7 years, height 171 +/- 9 cm, weight 64.2 +/- 9.0 kg, body fat 14.6 +/- 5.5%) completed the study. Subjects started maximum effort trials that were either hydrated (HYR) and dehydrated (DHR). Each trial subjects ran three 4-km loops with a 4-minute rest between loops. Significance was set at p < or = 0.05. The DHR had a significantly greater body mass loss at the pre- and posttrial time points (-2.05 +/- 1.25 and -4.3 +/- 1.25%, respectively) vs. HYR (-0.79 +/- 0.95 and -2.05 +/- 1.09%, respectively). Subjects ran the 12 km faster (p < 0.001) in HYR (3,191 +/- 366 seconds) vs. DHR (3,339 +/- 450 seconds). Differences between fastest and slowest loops during HYR (54 +/- 40 seconds) were significantly smaller than DHR (111 +/- 93 seconds; p = 0.041). Additionally, loop times were slower for loop 1 (HYR 1,039 +/- 116 seconds vs. DHR 1,071 +/- 123 seconds; p = 0.028), loop 2 (HYR 1,066 +/- 123 seconds vs. DHR 1,105 +/- 148 seconds; p = 0.01) and loop 3 (HYR 1,081 +/- 132 seconds vs. DHR 1,168 +/- 189 seconds; p = 0.003) when dehydrated. Percent of the race completed by loop as calculated by finishing time was significantly different at loop 2 between HYR (33.6 +/- 0.36%) and DHR (33.1 +/- 0.35%, p = 0.002) and loop 3 (33.8 +/- 0.75% vs. 34.9 +/- 1.35%, respectively, p = 0.01). Total variation from the mean pace for the duration of the HYR compared to the DHR approached significance (p = 0.064). Average percent of variance approached significance between trials (p = 0.057). Differences between the fastest and slowest loops between trials demonstrated an increased ability for hydrated individuals to evenly pace themselves. While total variation from the mean pace was not significantly different, it could have practical applicability. These findings reveal that dehydration is associated with decreases in a runners' ability to evenly pace themselves during a competitive situation. PMID- 19675478 TI - Validity of the running anaerobic sprint test for assessing anaerobic power and predicting short-distance performances. AB - The purpose of this study was to investigate the reliability and validity of the running anaerobic sprint test (RAST) in anaerobic assessment and predicting short distance performance. Forty members of the armed forces were recruited for this study (age 19.78 +/- 1.18 years; body mass 70.34 +/- 8.10 kg; height 1.76 +/- 0.53 m; body fat 15.30 +/- 5.65 %). The RAST test was applied to six 35-meter maximal running performances with a 10-second recovery between each run; the peak power, mean power, and the fatigue index were measured. The study was divided in two stages. The first stage investigated the reliability of the RAST using a test retest method; the second stage aimed to evaluate the validity of the RAST comparing the results with the Wingate test and running performances of 35, 50, 100, 200, and 400 m. There were not significant differences between test-retest scores in the first stage of the study (p > 0.05) and were found significant correlations between these variables (intraclass correlation coefficient approximately = 0.88). The RAST had significant correlations with the Wingate test (peak power r = 0.46; mean power r = 0.53; fatigue index r = 0.63) and 35, 50, 100, 200, and 400 m performances scores (p < 0.05). The advantage of using the RAST for measuring anaerobic power is that it allows for the execution of movements more specific to sporting events that use running as the principal style of locomotion, is easily applied and low cost, and due to its simplicity can easily be incorporated into routine training. We concluded that this procedure is reliable and valid, and can be used to measure running anaerobic power and predict short-distance performances. PMID- 19675480 TI - Effect of circuit training on the sprint-agility and anaerobic endurance. AB - The purpose of this study was to determine the effect of circuit training directed toward motion and action velocity over the sprint-agility and anaerobic endurance. A total of 32 healthy male physical education students with a mean age of 23.92 +/- 1.51 years were randomly allocated into a circuit training group (CTG; n = 16) and control group (CG; n = 16). A circuit training consisting of 8 stations was applied to the subjects 3 days a week for 10 weeks. Circuit training program was executed with 75% of maximal motion numbers in each station. The FIFA Medical Assessment and Research Centre (F-MARC) test battery, which was designed by FIFA, was used for measuring sprint-agility and anaerobic endurance. Pre- and posttraining testing of participants included assessments of sprint-agility and anaerobic endurance. Following training, there was a significant (p < 0.05) difference in sprint-agility between pre- and posttesting for the CTG (pretest = 14.76 +/- 0.48 seconds, posttest = 14.47 +/- 0.43 seconds). Also, there was a significant (p < 0.05) difference in anaerobic endurance between pre- and posttesting for the CG (pretest = 31.53 +/- 0.48 seconds, posttest = 30.73 +/- 0.50 seconds). In conclusion, circuit training, which is designed to be performed 3 days a week during 10 weeks of training, improves sprint-agility and anaerobic endurance. PMID- 19675479 TI - Acute effects of dynamic stretching, static stretching, and light aerobic activity on muscular performance in women. AB - The purpose of this study was to compare three warm-up protocols--static stretching, dynamic stretching, and light aerobic activity--on selected measures of range of motion and power in untrained females and to investigate the sustained effects at 5 and 30 minutes after warm-up. A total of 24 healthy females (ages 23-29 years) attended one familiarization session and three test sessions on nonconsecutive days within 2 weeks. A within-subject design protocol with the testing investigators blinded to the subjects' warm-up was followed. Each session started with 5 minutes of light aerobic cycling followed by pretest baseline measures. Another 5 minutes of light aerobic cycling was completed and followed by one of the three randomly selected warm-up interventions (static stretching, dynamic stretching, or light aerobic activity). The following posttest outcome measures were collected 5 and 30 minutes following the intervention: modified Thomas test, countermovement jump, and isometric time to peak force knee extension measured by dynamometer. Analysis of the data revealed significant time effects on range of motion and countermovement jump changes. No significant differences (p > 0.05) were found between the warm-up conditions on any of the variables. The variation in responses to warm-up conditions emphasizes the unique nature of individual reactions to different warm-ups; however, there was a tendency for warm-ups with an active component to have beneficial effects. The data suggests dynamic stretching has greater applicability to enhance performance on power outcomes compared to static stretching. PMID- 19675481 TI - The effects of recovery interventions on consecutive days of intermittent sprint exercise. AB - The purpose of this study was to compare four recovery interventions following simulated team sport, intermittent-sprint exercise on consecutive days. Ten female netball players performed four randomized sessions of a simulated netball exercise circuit on consecutive days. Each condition consisted of two identical sessions (Session 1 and 2), with the recovery intervention implemented at the completion of Session 1. Participants performed all interventions involving: passive recovery, active recovery (ACT), cold water immersion (CWI) and contrast water therapy (C(T)WT). No significant differences (p > 0.05) were evident between conditions for exercise performance (vertical jump, 20-m sprint, 10-m sprint, total circuit time) during Session 2. Effect size data indicated trends for an ameliorated decline in 5 x 20-m sprints and vertical jump for C(T)WT and CWI, respectively. C(T)WT demonstrated a significant reduction (p = 0.04) in lactate post-intervention compared to ACT recovery. Further, ACT recovery resulted in a significantly elevated (p < 0.01) heart rate compared to all other conditions postintervention and demonstrated significantly higher (p < 0.01) rating of perceived exertion postintervention and muscle soreness pre-exercise Session 2. It is likely that while interventions may be applicable to team sport practices, the 24-hour recovery period between exercise bouts was sufficient to allow performance to be maintained, regardless of recovery interventions. PMID- 19675482 TI - The effects of compression garments on recovery. AB - The purpose of this study was to investigate whether wearing lower-body compression garments attenuate indices of muscle damage and decrements in performance following drop-jump training. Seven trained female and four trained male subjects undertook blood collection for creatine kinase (CK) and lactate dehydrogenase (LDH), a mid-thigh girth measurement, and reported their perceived muscle soreness (PMS). A series of performance tests were then completed including sprints (5 m, 10 m, and 20 m), a 5-0-5 agility test, and a countermovement jump test. In a randomized crossover experimental design, separated by 1 week, subjects completed 5 x 20 maximal drop-jumps, followed immediately after exercise by either wearing graduated compression tights (CG) or undertook passive recovery as a control (CON) for 48 hours. CK, LDH, mid-thigh girth, and PMS were retested after 24 hours and 48 hours of recovery. The performance tests were repeated after 48 hours of recovery. Analysis of variance for repeated measures indicated that for female subjects, CK values were elevated after 24-hour recovery (p = 0.020) and a greater PMS was observed after 48-hour recovery in the CON condition (p = 0.002) but not for the CG condition. For all the subjects (n = 11), a greater PMS was observed after 48-hour recovery in the CON condition (p = 0.001) but not the CG condition. Significant increases in time were reported for 10-m (p = 0.016, 0.004) and 20-m sprints (p = 0.004, 0.001) in both the CON and CG conditions and for the 5-m sprint (p = 0.014) in the CG condition. All other parameters were unchanged in either condition. Data indicates that CK responses and PMS might be attenuated by wearing compression tights in some participants after drop-jump training; however, no benefit in performance was observed. PMID- 19675483 TI - Complex training in professional rugby players: influence of recovery time on upper-body power output. AB - After a bout of heavy resistance training (HRT), skeletal muscle is in both a fatigued and potentiated state. Subsequent muscle performance depends on the balance between these 2 factors. To date, there is no uniform agreement about the recovery time required between the HRT and subsequent muscle performance to gain performance benefits in the upper body. The aim of the present study was to determine the recovery time required to observe enhanced upper-body muscle performance after HRT (i.e., complex training). Twenty-six professional rugby players performed a ballistic bench press (BBP) at baseline and at approximately 15 seconds and 4, 8, 12, 16, 20, and 24 minutes after HRT (3 sets of 3 repetitions at 87% 1 repetition maximum). Peak power output (PPO) and throw height were determined for all BBPs. A significant time effect with regard to PPO (F = 29.145, partial Eta2 = 0.538, p < 0.01) and throw height (F = 17.362, partial Eta2 = 0.410, p < 0.01) was observed. Paired comparisons indicated a significant decrease in PPO and throw height in the BBP performed approximately 15 seconds after the HRT compared with the baseline BBP. After 8 minutes of recovery from the HRT, both PPO and throw height were significantly higher than the PPO and throw height recorded at baseline (e.g., PPO: 879 +/- 100 vs. 916 +/- 116 W, p < 0.01). It was concluded that muscle performance can be significantly enhanced after bouts of HRT during a BBP providing that adequate recovery (8 min) is given between the HRT and the explosive activity. PMID- 19675484 TI - Muscular power, neuromuscular activation, and performance in shot put athletes at preseason and at competition period. AB - The aim of this study was to investigate changes in shot put performance, muscular power, and neuromuscular activation of the lower extremities, between the preseason and the competition period, in skilled shot put athletes using the rotational technique. Shot put performance was assessed at the start of the pre season period as well as after 12 weeks, at the competition period, in nine shot putters. Electromyographic (EMG) activity of the right vastus lateralis muscle was recorded during all shot put trials. Maximum squat strength (1RM) and mechanical parameters during the countermovement jump (CMJ) on a force platform were also determined at pre-season and at competition period. Shot put performance increased 4.7% (p < 0.05), while 1RM squat increased 6.5% (p < 0.025). EMG activity during the delivery phase was increased significantly (p < 0.025) after the training period. Shot put performance was significantly related with muscular power and takeoff velocity during the CMJ, at competition period (r = 0.66, p < 0.05 and 0.70, p < 0.05), but not with maximum vertical force. One RM squat was not related significantly with shot put performance. These results suggest that muscular power of the lower extremities is a better predictor of rotational shot put performance than absolute muscular strength in skilled athletes, at least during the competition period. PMID- 19675485 TI - Training with independent cranks alters muscle coordination pattern in cyclists. AB - In cycling, a circular pedaling action makes the most useful contribution to forward propulsion. Training with independent cranks (IC) has been proposed to improve the pedaling action. The aims of this study were, first, to assess whether the intermuscular coordination pattern of the pedaling action with normal cranks (NC) is modified after a training period with IC and, second, to determine if the new coordination pattern is maintained after a washing-out period. Eighteen cyclists, divided into a control (CG) and an experimental (EG) group, underwent 2 test sessions (T1 and T2) separated by 2 weeks of training (18 hours). The electromyographic (EMG) activity of 4 lower limbs' muscles was recorded while the athletes pedaled at 80 rpm for 60 seconds at 30 and 50% of the maximal power output determined during a maximal pedaling test. The tasks were performed with IC (EG) and NC (EG and CG). The EG underwent a retention test session (T3) after another 18-hour training with NC. EG showed a significant (45.8 +/- 8.8 vs. 36.0 +/- 6.1%, p < 0.01 at 30% intensity) and a quasi significant (62.7 +/- 10.3 vs. 54.2 +/- 8.7%, p = 0.09 at 50% intensity) decrease in vastus lateralis EMG activity and a quasi-significant (36.4 +/- 13.4 vs. 43.5 +/- 10.9%, p = 0.09 at 30% intensity) and a significant (54.5 +/- 12.1 vs. 65.5 +/- 16.1%, p < 0.05 at 50% intensity) increase in biceps femoris EMG activity between T1-NC and T2-NC. By T3, EMG activity returned to initial levels (T1). On the contrary, CG did not reveal any significant variation. The results provide scientific support for muscle coordination pattern alteration from the use of IC, potentially achieving a more effective pedaling action. IC training reduces quadriceps exertion, thus preserving it for important moments during competition. PMID- 19675486 TI - Effects of low- vs. high-cadence interval training on cycling performance. AB - High-resistance interval training produces substantial gains in sprint and endurance performance of cyclists in the competitive phase of a season. Here, we report the effect of changing the cadence of the intervals. We randomized 18 road cyclists to 2 groups for 4 weeks of training. Both groups replaced part of their usual training with 8 30-minute sessions consisting of sets of explosive single leg jumps alternating with sets of high-intensity cycling sprints performed at either low cadence (60-70 min(-1)) or high cadence (110-120 min(-1)) on a training ergometer. Testosterone concentration was assayed in saliva samples collected before and after each session. Cycle ergometry before and after the intervention provided measures of performance (mean power in a 60-s time trial, incremental peak power, 4-mM lactate power) and physiologic indices of endurance performance (maximum oxygen uptake, exercise economy, fractional utilization of maximum oxygen uptake). Testosterone concentration in each session increased by 97% +/- 39% (mean +/- between-subject SD) in the low-cadence group but by only 62% +/- 23% in the high-cadence group. Performance in the low-cadence group improved more than in the high-cadence group, with mean differences of 2.5% (90% confidence limits, +/-4.8%) for 60-second mean power, 3.6% (+/-3.7%) for peak power, and 7.0% (+/-5.9%) for 4-mM lactate power. Maximum oxygen uptake showed a corresponding mean difference of 3.2% (+/-4.2%), but differences for other physiologic indices were unclear. Correlations between changes in performance and physiology were also unclear. Low-cadence interval training is probably more effective than high-cadence training in improving performance of well-trained competitive cyclists. The effects on performance may be related to training associated effects on testosterone and to effects on maximum oxygen uptake. PMID- 19675487 TI - Determination of the ventilatory threshold with affective valence and perceived exertion in trained cyclists: a preliminary study. AB - The aim of the present study was to determine if trained cyclists have perceptual markers that can be used to identify the ventilatory threshold during a maximal exercise test. Ten trained cyclists (age 25.20 +/- 4.94 years; VO2max 63.6 +/- 6.3 mL x min(-1) x kg(-1)) performed a maximal graded test on a cycling ergometer. Perceived exertion and affective valence were measured each min of the maximal graded test. The results indicate that ventilatory threshold, perceived exertion, and affective valence are significantly different compared to the values assessed 1 minute after ventilatory threshold. Moreover, there were a significant relationships between power output and perceived exertion (p < 0.01; r = 0.97) and affective valence (p < 0.01; r = 0.94) measured during the maximal graded test. These results confirm that at ventilatory threshold a lot of significant changes of perceptual parameters appear and may be good indicators for athletes to determine their ventilatory threshold during a training session. This method could be interesting for cyclists to control the exercise intensity during individual time trial. PMID- 19675488 TI - Prediction VO2max during cycle ergometry based on submaximal ventilatory indicators. AB - There are several equations to predict maximum oxygen consumption (VO2max) from ergometric test variables on different ergometers. However, a similar equation using ventilatory thresholds of ergospirometry in a submaximal test on a cycle ergometer is unavailable. The aim of the present study was to assess the accuracy of VO2max prediction models based on indicators of submaximal effort. Accordingly, 4,640 healthy, nonathlete women ages 20 years and older volunteered to be tested on a cycle ergometer using a maximum incremental protocol. The subjects were randomly assigned to 2 groups: group A (estimation) and group B (validation). From the independent variables of weight in kilograms, the second workload threshold (WT2), and heart rate of the second threshold (HRT2), it was possible to build a multiple linear regression model to predict maximal oxygen consumption (VO2max = 40.302 - 0.497 [Weight] - 0.001 [HRT2] + 0.239 [WT2] in mL O2/kg/min(-1); r = 0.995 and standard error of the estimate [SEE] = 0.68 mL O2/kg/min(-1)). The cross-validation method was used in group B with group A serving as the basis for building the model and the validation dataset. The results showed that, in healthy nonathlete women, it is possible to predict VO2max with a minimum of error (SEE = 1.00%) from submaximal indicators obtained in an incremental test. PMID- 19675489 TI - Effects of reduced training and detraining on upper and lower body explosive strength in adolescent male basketball players. AB - The purpose of this study was to assess and compare the effects of detraining and of a reduced training program on upper- and lower-body explosive strength in adolescent male basketball players. To study this, 15 subjects, aged 14 to 15 years old, were randomly assigned to 1 of the 2 following groups: reduced training (RT; n = 8) and detraining (DTR; n = 7). The participants were assessed on squat jump (SJ), countermovement jump (CMJ), Abalakov test (ABA), depth jump (DJ), mechanical power (MP), and medicine ball throw (MBT) after a 10-week in season complex training program (T0) and at the end of 4 (T4), 8 (T8), 12 (T12), and 16 (T16) weeks of detraining and of the reduced training periods. Both groups showed maintenance of explosive strength values and statistical similarity between them whatever the moment of evaluation. In conclusion, 16 weeks of detraining or of reduced training allow for the maintenance of the gains previously achieved by the application of a 10-week in-season complex training program. However, the lack of differences between detraining and reduced training leads to the conclusion that regular basketball practice can sustain by itself the previously achieved explosive strength gains, considering its mainly explosive characteristics. PMID- 19675490 TI - Computer simulations assessing the potential performance benefit of a final increase in training during pre-event taper. AB - A nonlinear model of training responses was utilized to test whether a 2-phase taper could be more effective than a traditional linear taper. Simulations were conducted using model parameters previously determined in 6 nonathletes trained on a cycle ergometer (non-ATH) and 7 elite swimmers trained in sport-specific conditions (ATH). Linear and 2-phase tapers were compared after a 28-day overload period at 120% of normal training. The 2-phase taper was assumed to be identical to the optimal linear taper, except for the final 3 days during which the training load was varied to maximize the final performance. The optimal linear taper was characterized by a mean training reduction by 32 +/- 6% during 35 +/- 6 days in non-ATH and by 49 +/- 18% during 33 +/- 16 days in ATH. The last 3 days of the 2-phase taper were characterized by a significant increase in training load by 23 +/- 18% in non-ATH and 29 +/- 42% in ATH (p < 0.005). The optimal taper characteristics were not statistically different between non-ATH and ATH. The maximal performance reached with the 2-phase taper was higher by 0.04 +/- 0.02% in non-ATH and 0.01 +/- 0.01% in ATH than with the optimal linear taper (p < 0.001). Positive and negative influences of training on performance were estimated as indicators of adaptation and fatigue, respectively. The negative influence was completely removed during both tapers, whereas the positive influence was slightly further enhanced during the 2-phase pattern. In conclusion, simulations showed that a 20 to 30% increase in training at the end of the taper, as compared to a prolonged reduction in training, allowed additional adaptations without compromising the removal of fatigue. PMID- 19675491 TI - Evaluating the heading in professional soccer players by playing positions. AB - The aim of this study is to evaluate the accuracy and coordination of heading in professional soccer players of different positions. A total of 243 professional soccer players were examined for the study. The Federation Internationale de Football Association Medical Assessment and Research Center (F-MARC) test battery was used to evaluate heading in soccer players. In the heading of the ball thrown from middle of the goal, forwards were better than goalkeepers (p < 0.05). There was not a significant statistical difference between defenders, midfielders, and forwards. In the heading of the ball thrown from the right side of the goalpost, defenders were better than goalkeepers (p < 0.05) and forwards were better than goalkeepers, defenders, and midfielders (p < 0.05). In conclusion, the reason for the heading of goalkeepers being worse than the others; goalkeepers use their hands in both training and matches, whereas for forwards and defensive players being better than goalkeepers depends mainly on their positions in soccer. Trainers and coaches should give place specific training programs to improve heading ability in the training schedule of soccer players at a professional level. PMID- 19675492 TI - Effects of lower-limb plyometric training on body composition, explosive strength, and kicking speed in female soccer players. AB - The aim of the present study was to examine how explosive strength, kicking speed, and body composition are affected by a 12-week plyometric training program in elite female soccer players. The hypothesis was that this program would increase the jumping ability and kicking speed and that these gains could be maintained by means of regular soccer training only. Twenty adult female players were divided into 2 groups: control group (CG, n = 10, age 23.0 +/- 3.2 yr) and plyometric group (PG, n = 10; age 22.8 +/- 2.1 yr). The intervention was carried out during the second part of the competitive season. Both groups performed technical and tactical training exercises and matches together. However, the CG followed the regular soccer physical conditioning program, which was replaced by a plyometric program for PG. Neither CG nor PG performed weight training. Plyometric training took place 3 days a week for 12 weeks including jumps over hurdles, drop jumps (DJ) in stands, or horizontal jumps. Body mass, body composition, countermovement jump height, DJ height, and kicking speed were measured on 4 separate occasions. The PG demonstrated significant increases (p < 0.05) in jumping ability after 6 weeks of training and in kicking speed after 12 weeks. There were no significant time x group interaction effects for body composition. It could be concluded that a 12-week plyometric program can improve explosive strength in female soccer players and that these improvements can be transferred to soccer kick performance in terms of ball speed. However, players need time to transfer these improvements in strength to the specific task. Regular soccer training can maintain the improvements from a plyometric training program for several weeks. PMID- 19675493 TI - Preseason physiological profile of soccer and basketball players in different divisions. AB - The purpose of this study was to examine and compare the cardiorespiratory performance and isokinetic muscle strength between Greek soccer and basketball players of different divisions before starting the training season. Study participants included 100 soccer players and 61 basketball players, who were assigned according to the kind of sport and division. All participants underwent anthropometric measurements and performed an exercise test on a treadmill to determine maximal oxygen uptake (VO2max). Peak torque for quadriceps and hamstring muscles was measured on isokinetic dynamometer at angular velocity of 60 degrees/s(-1), 180 degrees/s(-1), and 300 degrees/s(-1). The statistical p value was set at p < 0.05. In soccer players VO2max in absolute and relative values was significantly lower in division IV compared to the other 3 divisions (3,413.4 +/- 351.0 vs. 3,932.7 +/- 551.2, 4,172.7 +/- 371.8, 4,223.0 +/- 323.8 ml x min(-1), respectively; p < 0.001 and 46.93 +/- 4.20 vs. 52.47 +/- 3.66, 54.86 +/- 3.80, 55.32 +/- 3.33 ml x kg x min(-1), respectively; p < 0.001). Basketball players presented significantly higher VO2max, in absolute values, compared to soccer players for divisions II (4,586.3 +/- 586.3 vs. 4,172.7 +/- 371.8 ml x min(-1); p < 0.05), III (4,319.6 +/- 418.6 vs. 3,932.7 +/- 551.2 ml x min(-1); p < 0.01), and IV (4,624.0 +/- 627.6 vs. 3,413.4 +/- 351.0 ml x min(-1); p < 0.001), respectively. Regarding peak torque, only basketball players showed significantly higher values at 60 degrees/sec(-1) in hamstrings for III (p < 0.05) and IV division (p < 0.05). Conclusively, the higher VO2max reached by professional soccer and basketball players compared to semiprofessional and amateur ones and between the soccer and basketball players of the same division can be attributed to the different duration of the maintenance period and to the effect of the training session on each sport, respectively. Finally, a higher level of muscle strength would be preferable in soccer and basketball and would reduce the risk for injuries in the maintenance and rebuilding training periods. PMID- 19675494 TI - The effect of a 4-week training regimen on body fat and aerobic capacity of professional soccer players during the transition period. AB - The purpose of this study was to evaluate the changes in body fat percentage and aerobic capacity in professional soccer players, after the implementation of a specific 4-week training regimen during the transition period. Fifty-eight professional soccer players of the Greek Premier National Division were separated in experimental (n = 38) and control groups (n = 20). Body composition and maximum oxygen intake were evaluated before and after a 4-week training regimen followed during the transition period. The experimental design used for analyzing weight (kg), percent body fat (%) and VO2 max values (ml x kg(-1) x min(-1)) was a 2 x 2 (Groups x Measures), with Groups as a between-subjects factor and Measures as a within-subjects factor. The level of significance was set at p < or = 0.05 for all analyses. Analyses of variances showed that the experimental and the control groups achieved statistically significant (a) increases from pretest to posttest measures in body weight (0.595 kg and 1.425 kg, respectively) and percent body fat (0.25 and 0.82, respectively), and (b) decreases in VO2 max values from pretest to posttest measures (0.81 and 3.56, respectively). The findings of the study revealed that the players who followed the training regimen compared with the players that did not follow any specific training program gained less weight and body fat and exhibited lower reduction in their VO2 max values. PMID- 19675495 TI - Are skinfold-based models accurate and suitable for assessing changes in body composition in highly trained athletes? AB - This study was designed to assess the usefulness of skinfold (SKF) equations developed by Jackson and Pollock (JP) and by Evans (Ev) in tracking body composition changes (relative fat mass [%FM], absolute fat mass [FM], and fat free mass [FFM]) of elite male judo athletes before a competition using a 4 compartment (4C) model as the reference method. A total of 18 male, top-level (age: 22.6 +/- 2.9 yr) athletes were evaluated at baseline (weight: 73.4 +/- 7.9 kg; %FM4C: 7.0 +/- 3.3%; FM4C: 5.1 +/- 2.6 kg; and FFM4C: 68.3 +/- 7.3 kg) and before a competition (weight: 72.7 +/- 7.5 kg; %FM4C: 6.5 +/- 3.4%; FM4C: 4.8 +/- 2.6 kg; and FFM4C: 67.9 +/- 7.1 kg). Measures of body density assessed by air displacement plethysmography, bone mineral content by dual energy X-ray absorptiometry, and total-body water by bioelectrical impedance spectroscopy were used to estimate 4C model %FM, FM, and FFM. Seven SKF site models using both JP and Ev were used to estimate %FM, FM, and FFM along with the simplified Ev3SKF site. Changes in %FM, FM, and FFM were not significantly different from the 4C model. The regression model for the SKF in question and the reference method did not differ from the line of identity in estimating changes in %FM, FM, and FFM. The limits of agreement were similar, ranging from -3.4 to 3.6 for %FM, -2.7 to 2.5 kg for FM, and -2.5 to 2.7 kg for FFM. Considering the similar performance of both 7SKF- and 3SKF-based equations compared with the criterion method, these data indicate that either the 7- or 3-site SFK models are not valid to detect %FM, FM, and FFM changes of highly trained athletes. These results highlighted the inaccuracy of anthropometric models in tracking desired changes in body composition of elite male judo athletes before a competition. PMID- 19675496 TI - Displacement and frequency for maximizing power output resulting from a bout of whole-body vibration. AB - Whole-body vibration (WBV) has been shown to be effective for increasing lower body power; however, the combination of frequency, displacement, and duration that elicits the best acute response has yet to be determined. The purpose of this study was to identify the protocol eliciting the greatest improvement in power after an acute bout of WBV. Forty men and women participated in this study, in which 8 different combinations of 30, 35, 40, and 50 Hz with 2-mm and 5-mm displacements were tested over 3 days. For all protocols, randomized to reduce potential order effects, subjects underwent 30 seconds of WBV while holding an isometric squat at a knee angle of 2.27 rad. Power was assessed by countermovement jumps. Subjects performed 3 jumps before WBV, immediately afterward, and 1, 5, and 10 minutes later. The highest normalized peak power (nPP) at each time point was determined using a 4 (frequency) x 2 (displacement) x 5 (time) repeated-measures analysis of variance. Significant effects were seen for frequency (p < or = 0.026) and time (p < or = .0001). Post hoc analyses revealed that the 30-Hz condition (1.010 +/- 0.003) produced a higher nPP than 35 Hz (1.00 +/- 0.003, p < or = 0.026) and 40 Hz (1.002 +/- 0.002, p < or = 0.028) but not 50 Hz (1.004 +/- .002). We also found a significantly higher nPP for the 1-minute post-treatment time point (1.011 +/- .0003) vs. all other time points (p < or = 0.006). Our data show that an acute WBV bout can significantly increase power output at 1 minute post-treatment across all frequencies and displacements, although 30 Hz appears to have a greater effect on power output than either 35 Hz or 40 Hz, but not 50 Hz, at all post-treatment time points. PMID- 19675497 TI - Effect of iTonic whole-body vibration on delayed-onset muscle soreness among untrained individuals. AB - Attempts to reduce or eliminate delayed-onset of muscle soreness are important as this condition is painful and debilitating. The purpose of this study was to examine the effectiveness of whole-body vibration (WBV) massage and stretching exercises at reducing perceived pain among untrained men. Sixteen adult men (age, 36.6 +/- 2.1 yr) volunteered to perform a strenuous exercise session consisting of resistance training and repeated sprints. Subjects were randomly assigned to 1 of 2 recovery groups: a group performing WBV stretching sessions or a stretching group performing static stretching without vibration. Both groups performed similar stretches, twice per day for 3 days after the workout. The vibration group performed their stretches on the iTonic platform (frequency, 35 Hz; amplitude, 2 mm). Perceived pain was measured at 12, 24, 48, and 72 hours postworkout. Statistical analyses identified a significantly lower level of reported perceived pain at all postworkout measurement times among the WBV group (p < 0.05). No difference existed at the preworkout measurement time. The degree of attenuation of pain ranged from 22-61%. These data suggest that incorporating WBV as a recovery/regeneration tool may be effective for reducing the pain of muscle soreness and tightness after strenuous training. PMID- 19675498 TI - Effects of load and contraction velocity during three-week biceps curls training on isometric and isokinetic performance. AB - The velocity-specificity principle in training is well established by studies applying isokinetic training devices. However, the contraction velocity during customary resistance training using barbells is rarely stable and can be manipulated in several ways. By manipulating load and intention of movement, the significance of contraction velocity during barbell training on gains in strength related parameters was investigated. Twenty-seven subjects (divided into 3 experimental groups) performed standardized biceps curls 3 times a week for 3 weeks under the following conditions: high load and slow contraction velocity (HS), high load and fast contraction velocity (HF), and low load with fast contraction velocity (LF). Twelve subjects received no intervention, serving as controls (C). Elbow flexion strength was tested before and after the training period at both isometric and 4 isokinetic contraction velocities (30, 90, 240 and 300 degrees/sec) using a dynamometer. Rate of force development (RFD) was calculated in 100 millisecond epochs from isometric torque curves. Increased maximal isometric strength was seen in HF (9.7%), whereas HS improved slow isokinetic strength (8.5%). There were no improvements in force performance for LF and C. In none of the groups were changes in RFD observed. These findings support the principle of training specificity, highlighting the importance of details concerning contraction velocity on the outcome of resistance training using free weights. PMID- 19675499 TI - Effects of four weeks of high-intensity interval training and creatine supplementation on critical power and anaerobic working capacity in college-aged men. AB - The critical power test provides 2 measures, critical power (CP) and anaerobic working capacity (AWC). In theory, the CP measurement represents the maximal power output that can be maintained without fatigue, and AWC is an estimate of work capacity associated with muscle energy reserves. High-intensity interval training (HIIT) has been shown to be an effective training method for improving endurance performance, including VO2PEAK. In addition, creatine (Cr) supplementation has been reported to improve AWC without training; however, it has shown no effect on CP. The purpose of this study was to examine the effects of 4 weeks of HIIT with Cr supplementation on CP and AWC. Forty-two recreationally active men volunteered to participate in this study. Participants were randomly assigned to 1 of 3 groups: Cr (n = 16), 10 g Cr + 10 g dextrose; placebo (PL, n = 16), 20 g dextrose; control (CON, n = 10), no treatment. Before and after supplementation, each participant performed a maximal oxygen consumption test VO2PEAK on a cycle ergometer to establish peak power output (PPO). Participants then completed a CP test involving 3 exercise bouts with the workloads set as a percentage of their PPO to determine CP and AWC. After a 2 week familiarization period of training and supplementing, PPO, CP, and AWC were remeasured before an additional 4 weeks of HIIT and supplementation were completed. Training consisted of 5 sets of 2-minute exercise bouts with 1 minute rest in between performed on the cycle ergometer, with intensities based on PPO. A significant improvement in CP was observed in the Cr group (6.72% +/- 2.54%), whereas PL showed no significant change (3.87% +/- 2.30%), and CON significantly decreased (6.27% +/- 2.38%). Furthermore, no changes in AWC were observed in any of the groups after treatment. The current findings suggest that Cr supplementation may enhance the effects of intense interval endurance training on endurance performance changes. PMID- 19675500 TI - The relationship between the drive for muscularity and muscle dysmorphia in male and female weight trainers. AB - Muscle dysmorphia is a form of body dysmorphic disorder in which individuals have a pathological preoccupation with their muscularity and, more specifically, an extreme fear that their bodies are too small. Relatively few empirical studies have been completed on muscle dysmorphia, and even fewer studies on the relationship between the drive for muscularity and muscle dysmorphia in men and women. The purpose of this research was to examine the relationship between the drive for muscularity and muscle dysmorphia in male (n = 55) and female (n = 59) recreational weight trainers. Results revealed that the behavior and diet subscales of the drive for muscularity significantly predicted muscle dysmorphia in males and females accounting for 69% and 46% of the total variance, respectively. Although the overall scores of muscle dysmorphia do not indicate clinical levels, these findings suggest that behaviors such as arranging one's schedule around his/her training regimen and dieting in order to gain muscle predict characteristics of muscle dysmorphia in men and women. PMID- 19675501 TI - Differences between arms and legs on position sense and joint reaction angle. AB - The purpose of the present investigation was to compare the arms and legs in relation to position sense and joint reaction angle. Position sense at 30 degrees, 45 degrees, and 60 degrees flexion as well as joint reaction angle to release from 20 degrees, 40 degrees, and 60 degrees flexion of the elbow and knee joint were evaluated in 12 healthy men. The measurements were performed over 3 consecutive days to minimize the error attributable to the variability of the measured parameters. During the assessment of position sense, subjects had to place their limb as closely as possible to the reference angle. The joint reaction angle was measured by a new test in which the subjects had to stop the fall of their limb as soon as possible after it was released. All measurements were performed in a common isokinetic dynamometer. The results of position sense showed that the arms were placed closer to the reference angle compared with the legs (1.3 degrees vs. 3.1 degrees on average for 3 angles, respectively; p < 0.05). The arms also exhibited faster reaction angle to release compared with legs (3.4 degrees vs. 6.3 degrees on average for 3 angles, respectively; p < 0.05). In conclusion, the ability of arms to perform more accurate and faster movements than legs may mainly be attributed to the higher number of muscle spindles and the lower innervation ratio of arms. An imbalance of the determined relationship between arms and legs in position sense and reaction angle may indicate a neuromuscular disturbance. PMID- 19675502 TI - Relative and absolute reliability of a modified agility T-test and its relationship with vertical jump and straight sprint. AB - The aims of this study were to evaluate the reliability of a modified agility T test (MAT) and to examine its relationship to the free countermovement jump (FCMJ) and the 10-m straight sprint (10mSS). In this new version, we preserved the same nature of displacement of the T-test but we reduced the total distance to cover. A total of 86 subjects (34 women: age = 22.6 +/- 1.4 years; weight = 63.7 +/- 10.2 kg; height = 1.65 +/- 0.05 m; body mass index = 23.3 +/- 3.3 kg x m(-2) and 52 men: age = 22.4 +/- 1.5 years; weight = 68.7 +/- 8.0 kg; height = 1.77 +/- 0.06 m; body mass index = 22.0 +/- 2.0 kg x m(-2)) performed MAT, T test, FCMJ, and 10mSS. Our results showed no difference between test-retest MAT scores. Intraclass reliability of the MAT was greater than 0.90 across the trials (0.92 and 0.95 for women and men, respectively). The mean difference (bias) +/- the 95% limits of agreement was 0.03 +/- 0.37 seconds for women and 0.03 +/- 0.33 seconds for men. MAT was correlated to the T-test (r = 0.79, p < 0.001 and r = 0.75, p < 0.001 for women and men, respectively). Significant correlations were found between both MAT and FCMJ, and MAT and 10mSS for women (r = -0.47, p < 0.01 and r = 0.34, p < 0.05, respectively). No significant correlations were found between MAT and all other tests for men. These results indicate that MAT is a reliable test to assess agility. The weak relationship between MAT and strength and straight speed suggests that agility requires other determinants of performance as coordination. Considering that field sports generally include sprints with change direction over short distance, MAT seems to be more specific than the T-test when assessing agility. PMID- 19675503 TI - The effect of local muscle endurance training on cardiorespiratory capacity in young women. AB - The purpose of this study was to investigate the effect of local muscle endurance training on maximal oxygen uptake and ventilatory threshold in young women. Nineteen untrained women, ranging in age from 18 to 26 years, were included in the study and assigned to two groups: the control group (n = 10), and the resistance training group (n = 9). The following variables were obtained at baseline and after 12 weeks: body mass; maximal oxygen uptake, maximal heart rate, maximal oxygen pulse, oxygen uptake at the ventilatory threshold, heart rate at the ventilatory threshold, and oxygen pulse at the ventilatory threshold assessed by cardiopulmonary exercise testing on treadmill; 1-repetition maximum (RM) tests in bench press, latissimus pull down, military press, lying barbell extension, standing barbell curls, leg press, knee extension, and hamstring curl. The training group underwent resistance strength training. Loading during training followed the concept of maximum repetitions. Each session was defined as the performance of three sets of 15RM with a 60-second rest between sets and exercises. No significant changes were observed in the control group before and after 12 weeks (p > 0.05). All 1RM tests increased after training (p < or = 0.01) in the training group, but no significant change was observed in body mass (p > 0.05). Cardiopulmonary variables showed no significant differences before and after resistance training (p > 0.05). These findings indicate that the local muscle endurance training realized produces no improvement in cardiorespiratory capacity in young women. PMID- 19675504 TI - Relationship between maximal squat strength and five, ten, and forty yard sprint times. AB - The purpose of this investigation was to examine the relationship between maximal squat strength and sprinting times. Seventeen Division I-AA male football athletes (height = 1.78 +/- 0.04 m, body mass [BM] = 85.9 +/- 8.8 kg, body mass index [BMI] = 27.0 +/- 2.6 kg/m2, 1 repetition maximum [1RM] = 166.5 +/- 34.1 kg, 1RM/BM = 1.94 +/- 0.33) participated in this investigation. Height, weight, and squat strength (1RM) were assessed on day 1. Within 1 week, 5, 10, and 40 yard sprint times were assessed. Squats were performed to a 70 degree knee angle and values expressed relative to each subject's BM. Sprints were performed on a standard outdoor track surface with timing gates placed at the previously mentioned distances. Statistically significant (p < or = 0.05) correlations were found between squat 1RM/BM and 40 yard sprint times (r = -0.605, p = 0.010, power = 0.747) and 10 yard sprint times (r = -0.544, p = 0.024, power = 0.626). The correlation approached significance between 5 yard sprint times and 1RM/BM (r = 0.4502, p = 0.0698, power = 0.4421). Subjects were then divided into those above 1RM/BM of 2.10 and below 1RM/BM of 1.90. Subjects with a 1RM/BM above 2.10 had statistically significantly lower sprint times at 10 and 40 yards in comparison with those subjects with a 1RM/BM ratio below 1.90. This investigation provides additional evidence of the possible importance of maximal squat strength relative to BM concerning sprinting capabilities in competitive athletes. PMID- 19675505 TI - A randomized controlled study of taper-down or abrupt discontinuation of hormone therapy in women treated for vasomotor symptoms. AB - OBJECTIVE: The aim of this study was to investigate whether tapering down of combined estrogen plus progestogen therapy (EPT) reduced the recurrence of hot flashes and resumption of therapy compared with abrupt discontinuation. A secondary aim was to evaluate whether health-related quality of life (HRQoL) was affected after discontinuation of EPT and to investigate the possible factors predicting resumption of EPT. METHODS: Eighty-one postmenopausal women undergoing EPT because of hot flashes were randomized to tapering down or abrupt discontinuation of EPT. Vasomotor symptoms were recorded in self-registered diaries, and resumption of hormone therapy (HT) was asked for at every follow-up. The Psychological General Well-being Index was used to assess HRQoL. RESULTS: Neither the number nor the severity of hot flashes or HRQoL or frequency of resumption of HT differed between the two modes of discontinuation of EPT during up to 12 months of follow-up. About every other woman had resumed HT within 1 year. Women who resumed HT after 4 or 12 months reported more deteriorated HRQoL and more severe hot flashes after discontinuation of therapy than did women who did not resume HT. CONCLUSIONS: Women who initiate EPT because of hot flashes may experience recurrence of vasomotor symptoms and impaired HRQoL after discontinuation of EPT regardless of the discontinuation method used, abrupt or taper down. Because, in addition to severity of flashes, decreased well-being was the main predictor of the risk to resume HT, it seems important to also discuss quality of life in parallel with efforts to discontinue HT. PMID- 19675506 TI - Daily vasomotor symptoms, sleep problems, and mood: using daily data to evaluate the domino hypothesis in middle-aged women. AB - OBJECTIVE: The aim of this study was to investigate the relationships among daily reports of vasomotor symptoms (hot flashes and/or sweats), same-day sleep problems, and next-day mood reports in middle-aged women. METHODS: Fifty-five healthy middle-aged women were recruited to keep daily records for up to 5 years or until menopause. For each participant, the first 252 days of contiguous data with the highest weekly frequency of vasomotor symptoms was selected for the current analyses. Multilevel structural equation modeling was used to test whether changes in daily vasomotor symptom occurrence predicted changes in occurrence of same-day sleep problems and changes in next-day positive and negative mood ratings and whether sleep problems mediated any predictive effect of symptoms on next-day mood. RESULTS: Controlling for initial depression, daily vasomotor symptoms predicted same-day sleep problems (b = 0.59, P < 0.001) and next-day positive mood (b = -0.07, P < 0.01), although significant direct relationships between vasomotor symptoms and mood were found primarily in women with initial depression scores in the low to moderate range. Sleep problems predicted next-day positive (b = -0.08, P < 0.01) and negative (b = 0.10, P < 0.001) mood more robustly than vasomotor symptoms did. CONCLUSIONS: Sleep problems predicted worse mood on the following day and accounted for only a small portion of the relationship between vasomotor symptoms and mood. These findings suggest that any effect of vasomotor symptoms on mood may occur largely through a mechanism other than sleep disruption. PMID- 19675507 TI - Cognitive change effects in a semantic, a numeric, and a figure inference task. AB - Event-related potentials were recorded to examine the electrophysiologic correlates of the evaluation of possible answers provided during a semantic, a numeric, and a figure inference task. We examined two conditions: the probe stimulus given was what participants initially generated (Correct answer) or it was unexpected and incorrect (Unexpected Incorrect answer). Results showed that Unexpected Incorrect answers elicited a more negative ERP deflection (N320) than did Correct answers between 300-400 ms under all inference tasks. The generator of N320 effect was localized near the anterior cingulate cortex. The N320 effect, therefore, reflects an expectation violation while participants performed semantic, numeric, or figure inference task. PMID- 19675508 TI - Developmental change of Na(+)-absorptive function in Reissner's membrane epithelia. AB - The purpose of this study was to investigate the developmental change of epithelial Na channel function in epithelial cells of rat Reissner's membrane. The Reissner's membrane was isolated from the upper turn of the cochlea at different stages of development. The vibrating probe technique was chosen to measure transepithelial currents under short circuit conditions. Amiloride sensitive Na current in the Reissner's membrane developed postnatally between days 3 and 5, which was completely blocked by amiloride. This tendency remained from days 5 through 14. Development of Na transport in epithelial cells of the Reissner's membrane may contribute considerably to the accomplishment of low Na concentration in the endolymph during early neonatal period. PMID- 19675509 TI - Ultrasound examination of the lungs in the intensive care unit. AB - OBJECTIVE: Lung ultrasound is increasingly used in the critically ill adult. It allows prompt management based upon reproducible data and generates fewer computed tomography (CT) examinations, therefore decreasing irradiation, delays, cost, and discomfort to the patient. The aim of this article is to describe the value of ultrasound for lung imaging in the critically ill and state our experience in neonates. METHODS: Review of studies published in the peer-reviewed international literature analyzing consecutive critically ill adults admitted to intensive care units, assessing pleural effusion, alveolar consolidation, interstitial syndrome, and pneumothorax, using a standardized ultrasound approach to the lung, with CT as the reference. DATA SYNTHESIS: The sensitivity and specificity of ultrasound are 92% and 93% for pleural effusion, 90% and 98% for alveolar consolidation, 93% and 93% for interstitial syndrome, 100% and 96% for complete pneumothorax, 79% and 100% for radio-occult pneumothorax. DISCUSSION: This article reviews data that validate the scientific value of lung ultrasound in adult medical intensive care units. We then present observations in the critically ill neonate. The discussion points to the methodologic issues raised in lung ultrasound in the neonate, i.e. mainly the limited access to a pertinent gold standard (CT). Some CT correlations are presented, confirming the value of lung ultrasound in the neonate. CONCLUSIONS: The standardized signs assessed in the adult are also found in the critically ill neonate, meaning a potential use in this field. Awaiting confirmatory CT studies, lung ultrasound can be taken into consideration as a possible bedside tool for completing bedside radiography. PMID- 19675510 TI - Trends in health care expenditures, utilization, and health status among US adults with spine problems, 1997-2006. AB - STUDY DESIGN: Analysis of nationally representative survey data for spine-related health care expenditures, utilization and self-reported health status. OBJECTIVE: To study trends from 1997 to 2006 in per-user expenditures for spine-related inpatient, outpatient, pharmacy, and emergency services; and to compare these trends to changes in health status. SUMMARY OF BACKGROUND DATA: Although prior work has shown overall spine-related expenditures accounted for $86 billion in 2005, increasing 65% since 1997, the study did not report per-user expenditures. Understanding population-level per-user expenditure for specific services relative to changes in the health status may help assess the value of these services. METHODS: We analyzed data from the Medical Expenditure Panel Survey, a multistage survey sample designed to produce unbiased national estimates of health care utilization and expenditure. Spine-related hospitalizations, outpatient visits, prescription medications and emergency department visits were identified using ICD-9-CM diagnosis codes. Regression analyses controlling for age, sex, comorbidity, and time (years) were used to examine trends from 1997 to 2006 in inflation-adjusted per-user expenditures, and utilization, and self reported health status. RESULTS: An average of 1774 respondents with spine problems was surveyed per year; the proportion suggested an increase in the number of people who sought treatment for spine problems in the United States from 14.8 million in 1997 to 21.9 million in 2006. From 1997 to 2006, the mean adjusted per-user expenditures were the largest component of increasing total costs for inpatient hospitalizations, prescription medications, andemergency department visits, increasing 37% (from $13,040 in 1997 to $17,909 in 2006), 139% (from $166 to $397), and 84% (from $81 to $149), respectively. A 49% increase in the number of patients seeking spine-related care (from 12.2 million in 1997 to 18.2 million in 2006) was the largest contributing factor to increased outpatient expenditures. Population measures of mental health and work, social, and physical limitations worsened over time among people with spine problems. CONCLUSION: Expenditure increases for spine-related inpatient, prescription, and emergency services were primarily the result of increasing per-user expenditures, while those related to outpatient visits were primarily due to an increase in the number of users of ambulatory services. PMID- 19675511 TI - Comparative study of neck pain in relation to increase of cervical epidural pressure during percutaneous endoscopic lumbar discectomy. AB - STUDY DESIGN: A prospective study. OBJECTIVE: By monitoring cervical epidural pressure (EP) changes throughout the procedure, we intend to discover the effect of percutaneous endoscopic lumbar discectomy (PELD) on cervical epidural pressure and whether there is a correlation between posterior neck pain and increased cervical EP, which is known to have a linear correlation with intracranial pressure (ICP). SUMMARY OF BACKGROUND DATA: Patients sometimes complain of posterior neck pain during PELD. Due to the massive irrigation fluid used during the procedure, the possibility of increased ICP as the cause is quite strong. METHODS: Twenty-eight patients undergoing PELD with ASA physical status 1 or 2 were enrolled in this study. In all patients, a cervical epidural catheter was placed at the C6-C7 level before the procedure and was connected to a pressure transducer. Cervical EPs were monitored continuously throughout the procedure. Initial stabilized EP (EP), EP at the time of neck pain, maximal EP during the procedure, and EP at the end of the procedure were checked. Neck pain onset time from the beginning of irrigation and total irrigation time were also checked. RESULTS: Of 28 patients, 8 patients complained of neck pain. Neck pain onset time from the beginning of irrigation was 35.6 +/- 11.3 (mean +/- SD) minutes. The EP at the time of neck pain (52.9 +/- 9.2 mm Hg) was significantly higher than the maximal EP in patients without neck pain (34.8 +/- 14.7 mm Hg). In all patients who complained of neck pain, the cervical EP at the time of neck pain showed pressures above 37 mm Hg. The maximal EP in those with neck pain (73.6 +/- 25.8 mm Hg) was also significantly higher than the EP in those without neck pain (34.8 +/- 14.7 mm Hg). In a correlation study, patients with higher maximal EPs had higher probabilities of having neck pain. In 6 of 8 patients, an abrupt increase in EP was observed after the onset of neck pain, while in the other 2 patients, the procedure ended just after neck pain appeared. CONCLUSION: Neck pain occurring during PELD is associated with a highly increased cervical EP generated by continuous infusion. PMID- 19675512 TI - Weight gain associated with atypical and typical antipsychotics during treatment of adolescent schizophrenic psychoses: A retrospective study. AB - OBJECTIVES: The aim of the study was to assess weight changes associated with certain atypical (AAP) and typical (TAP) antipsychotic drugs in patients with early-onset schizophrenia and other related psychotic disorders. METHODS: Our retrospective study included 109 patients (52 boys, 57 girls) with a mean age of 15.8 +/- 1.6 years. The patients were evaluated based to their medical records prior to starting therapy, and then after 1, 3, and 6 weeks of treatment. RESULTS: During the first week of treatment, the AAP group (n = 85; risperidone, olanzapine, ziprasidone, and clozapine) gained 1.5% of baseline weight whereas the TAP group (n = 24; haloperidol, perphenazine, and sulpiride) gained only 0.2% (p = 0.049). Differences in relative changes between the two groups were not significant at weeks 3 and 6. Expressed as absolute values, patients in our sample gained an average of 3.4 kg (SD 3.2) on AAP and 2.0 kg (SD 3.9) on TAP during 6 weeks of treatment (p = 0.335). Only the risperidone, olanzapine, and clozapine groups had sufficient numbers of patients to allow a comparison at the endpoint of the study (week 6). The patients gained, on average, 3.6 kg (SD 2.6) on risperidone, 4.4 kg (SD 2.5) on olanzapine, and 2.1 kg (SD 4.0) on clozapine during the six weeks of treatment (p = 0.286). CONCLUSIONS: In our study, we did not find a difference in weight gain between the AAP and TAP groups, as large as has been described in the literature. It also seems plausible that the unique and variable weight changes associated with individual AAPs in the pediatric population are different from those observed in the adult population. PMID- 19675513 TI - Synergistic interaction between rilmenidine and ibuprofen in the writhing test in mice. AB - OBJECTIVES: The aim of the study was to ascertain whether rilmenidine, a second generation imidazoline-alpha-2-adrenoreceptor agonist, is able to increase analgesic effects of ibuprofen in the writhing test in mice. Experimental studies combining these agents have not yet been published. METHODS: An acetic acid (0.7%) solution was injected into the peritoneal cavity and the number of writhes was counted. The influence on locomotor performance was tested using the rotarod test. RESULTS: Rilmenidine, ibuprofen, and rilmenidine-ibuprofen fixed-ratio combinations produced dose-dependent antinociceptive effects. ED50 values were estimated for the individual drugs and an isobologram was constructed. The derived theoretical additive ED50 value for the rilmenidine-ibuprofen combination was 34.00 +/- 9.39 mg/kg. This value was significantly greater than the observed ED50 value which was 18.07 +/- 5.41 mg/kg, indicating a synergistic interaction. Rilmenidine did not impair motor coordination, as measured by the rotarod test, at antinociceptive and higher doses. CONCLUSIONS: The present results suggest that rilmenidine enhances the analgesic activity of ibuprofen. If rilmenidine produces antinociception in humans, then the synergistic antinociception of rilmenidine with ibuprofen could offer therapeutic advantage for clinical treatment of pain. PMID- 19675514 TI - Serum ghrelin levels and disturbances of the lipid profile in patients with acromegaly. AB - It is unknown if altered ghrelin secretion might contribute to the development of metabolic complications in acromegaly. The AIM OF THE STUDY was to: 1) assess if serum concentrations of total and acylated ghrelin in patients with acromegaly differ in the presence of various metabolic complications (hypercholesterolemia, hyperinsulinemia, hyperglicemia). 2) assess the correlations between concentrations of ghrelin and concentrations of GH, IGF-1, cholesterol, insulin and glucose in patients with acromegaly. MATERIALS: 24 patients with previously diagnosed acromegaly (11 subjects with active and 13 subjects with inactive disease) and 12 healthy subjects. 23 subjects were treated in the past with neurosurgery, 3 subjects with radiotherapy. 7 patients were receiving octreotide LAR at the time of the study. METHODS: In all studied subjects the concentrations of total ghrelin, acylated ghrelin, GH, IGF-1, insulin, glucose, total cholesterol, LDL cholesterol, HDL cholesterol, triglicerydes, were measured. RESULTS: The concentrations of total and acylated ghrelin did not significantly differ between patients with active and inactive disease. The mean concentrations of total and acylated ghrelin were significantly higher in acromegalic patients who presented with hypercholesterolemia compared with patients with normocholesterolemia. In patients with hypercholesterolemia the ratio of acylated/total ghrelin was 16%. In patients with active acromegaly there was a statistically significant positive correlation between the concentration of total ghrelin and the concentration of total cholesterol and LDL cholesterol. There was also a positive correlation between the concentration of acylated ghrelin and LDL cholesterol (without statistical significance). In patients with inactive acromegaly there was a statistically significant positive correlation between the concentration of acylated ghrelin and the concentration of triglycerides and a positive correlation between total ghrelin and triglycerides, but statistically insignificant. There were no differences in ghrelin levels depending on the insulin and glucose concentrations. CONCLUSIONS: Some metabolic complications of the disease might result not only from GH hypersecretion but also from altered ghrelin secretion. PMID- 19675515 TI - Radioiodine therapy in patients with amiodarone-induced thyrotoxicosis (AIT). AB - INTRODUCTION: Amiodarone (AM) is frequently used in the therapy of patients with cardiac disorders. However, due to high iodine content, it has side effects on thyroid function. The use of radioiodine therapy (RIT) in amiodarone-induced thyrotoxicosis (AIT) with low radioactive iodine uptake (RAIU) is still controversial. In these patients therapeutic choices for refractory disease include surgery, antithyroid drugs, or glu ocorticosteriods. AIM: The aim of the study was to evaluate the efficacy of RIT in patients presenting AIT and low RAIU in two-year follow-up. PATIENTS AND METHODS: 40 patients (25 men and 15 women) aged from 63 to 83 years (x +/- SD: 66.2 +/- 5.0 years; median: 65 years) treated with RIT were included into the study. In these patients AM therapy was essential for the underlying heart disorder, while surgery, antithyroid drugs or glucocorticosteroids, were contraindicated. Forty seven patients with toxic multinodular goiter (TMNG) (39 women and 8 men), matched for age (67 +/- 12 yr; range 54-89 yr), were enrolled into the study as a comparative group. The diagnostic procedures included baseline thyroid function tests (thyrothropin - TSH, free triiodothyronine - fT3 and free thyroxine - fT4 levels), thyroid autoantibodies measurement (antithyroglobulin autoantibodies - TgAb, antithyroid peroxidase autoantibodies - TPOAb, anti-TSH receptor autoantibodies - TRAb), thyroid ultrasonography, thyroid scintiscan and RAIU assessment. RESULTS: Serum values of TSH, TgAb, TPOAb and TRAb were undetectable in both groups. In patients with AIT fT4 level was 18.7 to 38.7 pmol/l (mean: 27.1 +/- 5.8) and fT3 concentration was 3.9 to 5.6 pmo/l (mean: 5.7 +/- 1.4), while in TMNG patients level of fT4 was 31.5 to 22.2 pmol/l (mean: 25,3 +/- 5,8) and fT3 concentration was 3.8 to 4,2 pmo/l (mean: 4,2 +/- 0,2). Mean RAIU values after 5h and 24h in AIT patients were 2.3 +/- 0.5 and 3.1 +/- 0.9%, while in TMNG patients were 18,0 +/- 3,8 and 35,7 +/- 9,1%, respectively. A significant difference (p<0.001) between 5h and 24h RAIU in AIT compared to TMNG was noted. In all patients with AIT, a dose of 800 MBq of 131I was administered. During two-year-observation recurrence of hyperthyroidism was observed in two patients (5%) with TMNG. These patients received a second radioiodine dose 16.2 +/- 15 months later (the mean re treatment dose was 735.93 +/- 196.1 MBq). In comparison, none of the patients with AIT required a second 131I dose and only one patient (2.5%) 6 months after ablative 131I dose needed anti-thyroid medication. Transient hypothyroidism was observed in only two patients (5%) with AIH, though was not observed in TMNG. During follow-up time, no sudden deaths in AIT patients were observed; one patient was diagnosed with prostate cancer, and in one patient acute toxic hepatitis after AM occurred. CONCLUSION: RIT may be a safe and useful method of AIT therapy in patients with low RAIU, in whom other treatment methods are contraindicated. PMID- 19675516 TI - Profiling of serum proteins influenced by warm partner contact in healthy couples. AB - OBJECTIVES: Warm physical contact may positively influence our health and well being; however, it has not been investigated yet whether serum proteins are influenced by warm physical contact in healthy couples. In this study, we focused on psychological and physiological effects of warm partner contact in healthy couples. METHODS: When participants freely kissed and hugged their romantic partners, they were asked to subjectively evaluate their present emotions. Furthermore, changes of serum proteins were determined by using ProteinChip surface enhanced laser desorption/ionization-time-of-flight-mass spectrometry (SELDI-TOF-MS). We characterized these proteins by using biochemical techniques combined with gel filtration high performance liquid chromatography (HPLC), reverse-phase HPLC, and sequencing analyses. RESULTS: Romantic couples became happier and less irritated after kissing and hugging. Accompanying these psychological changes, SELDI-TOF-MS indicated that the intensities of 66-k Da, 11.7-k Da, and 5.9-k Da serum proteins were increased. These proteins were identified as serum albumin and beta2-microglobulin, and probably fibrinogen fragment. The feeling of happiness positively correlated and the feeling of irritation negatively correlated with intensities of serum albumin and beta2 microglobulin. CONCLUSION: These results suggest that psychological stress may be reduced and we may feel happiness when we kiss and hug a romantic partner. Furthermore, these results also suggest that warm partner contact influences peripheral circulating proteins, more importantly, may promote health and well being. PMID- 19675517 TI - Modulation of thermal pain perception by stress and sweet taste in women with bulimia nervosa. AB - OBJECTIVES: To investigate if the increased pain threshold in women with bulimia nervosa (BN) may be due to chronic stress-induced analgesia. METHODS: We measured thermal pain threshold latency, blood pressure and heart rate in 21 women with BN and 21 healthy women (HW) under six consecutive conditions: rest I, mental arithmetic task, rest II, eating sweet food, rest III, cold-pressor test. RESULTS: Thermal pain threshold latency was longer in BN than in HW in all six conditions. It increased during mental arithmetic test and remained increased during the rest of the experiment in both groups. In the BN group, the increase of pain threshold during mental arithmetic was positively correlated with illness duration. The differential modulation of pain threshold by stress in BN and HW could not be explained by autonomic system reactivity. In HW, the pain threshold increased more during eating and blood pressure increased more during mental stress; in BN, the pain threshold was highest in the mental stress condition and blood pressure was most increased during eating. During the cold pressor test, women with BN showed smaller blood pressure increase and tolerated the cold for shorter time than HW. CONCLUSION: The observed marked modulation of pain threshold by experimental stress suggests that stress-induced analgesia is unlikely to account for baseline pain insensitivity in BN. Increased pain threshold in BN is a stable yet incompletely understood phenomenon, which may be related to the predisposition to or maintenance of the disorder. PMID- 19675518 TI - Meigs' syndrome and virilizing ovarian fibrothecoma complicating pregnancy. A case report and review of the literature. AB - BACKGROUND: A half of virilizing ovarian tumors in pregnancy are malignant. The risk of cancer increases with ascites and hydrothorax. Our case shows that such tumor can be benign with good outcome. CASE: Primigravida in 30 weeks with advanced hirsutism (from 15 weeks), vomiting, epigastric pain, dyspnoea, ascites, hydrothorax, ovarian mass and high level of serum testosterone was operated. After the tumorectomy (fibrothecoma) we performed cesarean section (placental abruption suspected). The female fetus had no signs of virilization. In mother all symptoms and effusions resolved. CONCLUSION: Meigs' syndrome in pregnancy leads to acute clinical symptoms and needs the surgical intervention with the risk of placental abruption. Virilization of mother was the first symptom of ovarian tumor but the female fetus was protected of hyperandrogenism. PMID- 19675519 TI - Endocannabinoid system: An overview of its potential in current medical practice. AB - The endocannabinoid system (ECS) is a lipid signalling system, comprising of the endogenous cannabis-like ligands (endocannabinoids) anandamide (AEA) and 2 arachidonoylglycerol (2-AG), which derive from arachidonic acid. These bind to a family of G-protein-coupled receptors, called CB1 and CB2. The cannabinoid receptor 1 (CB1R) is distributed in brain areas associated with motor control, emotional responses, motivated behaviour and energy homeostasis. In the periphery, the same receptor is expressed in the adipose tissue, pancreas, liver, GI tract, skeletal muscles, heart and the reproduction system. The CB2R is mainly expressed in the immune system regulating its functions. Endocannabinoids are synthesized and released upon demand in a receptor-dependent way. They act as retrograde signalling messengers in GABAergic and glutamatergic synapses and as modulators of postsynaptic transmission, interacting with other neurotransmitters. Endocannabinoids are transported into cells by a specific uptake system and degraded by the enzymes fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL). The ECS is involved in various pathophysiological conditions in central and peripheral tissues. It is implicated in the hormonal regulation of food intake, cardiovascular, gastrointestinal, immune, behavioral, antiproliferative and mammalian reproduction functions. Recent advances have correlated the ECS with drug addiction and alcoholism. The growing number of preclinical and clinical data on ECS modulators is bound to result in novel therapeutic approaches for a number of diseases currently treated inadequately. The ECS dysregulation has been correlated to obesity and metabolic syndrome pathogenesis. Rimonabant is the first CB1 blocker launched to treat cardiometabolic risk factors in obese and overweight patients. Phase III clinical trials showed the drug's ability to regulate intra-abdominal fat tissue levels, lipidemic, glycemic and inflammatory parameters. However, safety conerns have led to its withrawal. The role of endocannabinoids in mammalian reproduction is an emerging research area given their implication in fertilization, preimplantation embryo and spermatogenesis. The relevant preclinical data on endocannabinoid signalling open up new perspectives as a target to improve infertility and reproductive health in humans. PMID- 19675520 TI - Effects of fulvestrant, an estrogen receptor antagonist, on MMQ cells and its mechanism. AB - OBJECTIVES: Unlike the successful endocrine therapy of breast cancers and other estrogen-dependent diseases, little is known about the effect of anti-estrogen treatment on pituitary tumors. Our objectives were to study the effect of fulvestrant, a new type anti-estrogen devoid of any agonistic activities, on prolactinoma cell line MMQ in vitro and its possible mechanisms. DESIGN: In the experiment, the prolactin concentration, proliferation and apoptosis of the MMQ cell were measured to investigate the anti-tumor effect of the fulvestrant. The expression of estrogen receptor (ESR) mRNA and protein and MAPK pathway-related proteins ERK1 and 2, JNK, and p38 were measured to investigate the possible mechanisms. RESULTS: Fulvestrant significantly inhibited prolactin secretion (up to 85.5%), decreased proliferation (IC50 = 32.4 nmol/l), and promoted apoptosis of the MMQ cells. CONCLUSIONS: The suppression was possibly mediated by inhibition of ESR mRNA expression, down-regulation of ESR expression and activation of MAPK pathway-related proteins. Thus, fulvestrant has suppressive effects on prolactinoma cells and its anti-tumor mechanism appears to be related to the inhibition of ESR and the MAPK pathway. PMID- 19675521 TI - Effects of feminizing surgery for ambiguous genitalia - a novel scale for evaluation of cosmetic and anatomical results. AB - INTRODUCTION: Ambiguous genitalia always present diagnostic and therapeutic problem. Long-term results of feminizing operations are unsatisfactory in some cases and reports from follow-up after feminizing genitoplasty are rare in the literature. Systematic studies are needed to evaluate ultimate function of all girls undergoing feminizing surgery. In our opinion, the lack of worldwide accepted scale for the assessment of long-term effects of feminizing genitoplasty, enabling the possibility of comparing outcomes between institutions and countries, may contribute to this deficit of reliable data. AIM: The aim of the study was the evaluation of the outcomes of surgical management of masculinization using a novel scale developed by the authors. MATERIAL AND METHODS: We examined 43 patients aged 3-24 years (mean age 15,4 years) operated due to ambiguous genitalia. Most of the patients were females with congenital adrenal hyperplasia (CAH) diagnosed in 38 of subjects (88.4%).The patients were operated at the age from 10 months - 15 years (mean age 4,5 years). Five patients had to be reoperated. The effects of surgical management in ambiguous genitalia were assessed using our own scale. We evaluated five anatomical and cosmetic parameters (general appearance, size of pudendal labia, symmetry of pudendal labia symmetry, size and position of clitoris, size of introitus and position of urethra opening), each of them was scored 0-2 points. RESULTS: In 36 of examined patients the result of the surgery was considered good, in 4 patients - satisfactory and in 3 - poor. The most common complication of feminizing genitoplasty in our patients was stenosed vagina (in 10 patients, 23.8%). Location of urinary coil in anterior wall of vagina was found in 6 patients (13.9%) and in 1 patient it was invisible (2.3%). Post-operational clitoral enlargement was found in 3 cases (6.9%). CONCLUSION: Taking into consideration controversial data about the results of feminizing genitoplasty, a standarized, well-defined and commonly accepted scale enabling comparison between methods and institutions is necessary. In our opinion scoring scale makes the evaluation more precise and the results are more comparable. PMID- 19675522 TI - Effect of the polymorphism in 5' UTR region of pig prolactin gene on prolactin gene expression and reproduction performance in the female pig. AB - OBJECTIVES: The recent genetics and molecular biology progress seems to be a fascinating challenge for the interdisciplinary studies on the effects of genetic changes in gene structure that causes the modification of physiological functions of many important proteins including hormones. Pig prolactin is one of the interesting hormones for this study. AIM OF THE STUDY: The aim of the study was to analyze the mutation in 5'UTR region of the pig prolactin (PRL) gene and to evaluate the effect of this polymorphism on changes in plasma prolactin concentration. RESULTS: It was found that only two individual groups of animals differed by the genotype in examined PRL gene locus - homozygote C/C and heterozygote C/T. PRL plasma concentration was 38.4 ng/ml (for C/T animals) or 42.7 ng/ml (for C/C animals). Animals with C/C genotyped exhibited a tendency to elevate PRL concentration as compared to the C/T group (p< 0.07). CONCLUSIONS: This research combines the genetic, molecular and, in vivo, physiological study which allows focus on the possible relationship between the gene polymorphism and physiological status of animal. PMID- 19675523 TI - Dynamic analysis of bone marrow mesenchymal stem cells migrating to pancreatic islets using coculture microfluidic chips: An accelerated migrating rate and better survival of pancreatic islets were revealed. AB - OBJECTIVES: A coculture microfluidic chip had been fabricated to investigate the dynamic process of bone marrow mesenchymal stem cells migrating to the pancreatic islets in vitro. METHODS: We fabricated a coculture microfluidic chip using standard procedures. On the chip, bone marrow mesenchymal stem cells and pancreatic islets were introduced respectively into two microchambers which could be connected by a traffic tunnel. Migration assay was performed along the tunnel, and the function of pancreatic islets was timely evaluated by analysis of insulin secretion in response to high-glucose-challenge. RESULTS: The results showed that some bioactive factors excreted by freshly isolated pancreatic islets could enhance the migrating rate of BM-MSCs. Besides, pancreatic islets also showed a better survival and function by coculturing with bone marrow mesenchymal stem cells. CONCLUSION: The results demonstrated that bone marrow mesenchymal stem cells had the ability to migrate to pancreatic islets and provide an apparent overall preservation for islet function. This microfluidic device was a potentially useful tool to mimic actual biological processes of bone marrow mesenchymal stem cells migrating to the pancreatic islets in vitro. PMID- 19675524 TI - Cultural neuroeconomics of intertemporal choice. AB - According to theories of cultural neuroscience, Westerners and Easterners may have distinct styles of cognition (e.g., different allocation of attention). Previous research has shown that Westerners and Easterners tend to utilize analytical and holistic cognitive styles, respectively. On the other hand, little is known regarding the cultural differences in neuroeconomic behavior. For instance, economic decisions may be affected by cultural differences in neurocomputational processing underlying attention; however, this area of neuroeconomics has been largely understudied. In the present paper, we attempt to bridge this gap by considering the links between the theory of cultural neuroscience and neuroeconomic theory of the role of attention in intertemporal choice. We predict that (i) Westerners are more impulsive and inconsistent in intertemporal choice in comparison to Easterners, and (ii) Westerners more steeply discount delayed monetary losses than Easterners. We examine these predictions by utilizing a novel temporal discounting model based on Tsallis' statistics (i.e. a q-exponential model). Our preliminary analysis of temporal discounting of gains and losses by Americans and Japanese confirmed the predictions from the cultural neuroeconomic theory. Future study directions, employing computational modeling via neural networks, are outlined and discussed. PMID- 19675525 TI - Adiponectin - a predictor of higher mortality in cardiovascular disease or a factor contributing to longer life? AB - Adiponectin, a protein secreted by adipocytes, has been recently found to be also secreted by cardiomyocytes. Adiponectin possesses several physiological functions including modulation of glucose metabolism and energy homeostasis. The interactions between adiponectin and metabolic parameters were found. Studies on aging humans revealed that enhanced adiponectin values are a distinctive feature of centenarians. Thus, this parameter may be considered as a prognostic factor of prolonged survival. However, it has also been demonstrated that high adiponectin levels may predict mortality in patients with cardiovascular disease (CVD). Several mechanisms of adiponectin elevation, both in the process of aging and in pathomechanism of CVD, are discussed in this paper. PMID- 19675526 TI - 17beta-estradiol and predegenerated nerve graft effect on hippocampal neurogenesis in adult female rats. AB - OBJECTIVES: The aim of present work was to examine estrogen influence on neurogenesis in the model of predegenerated peripheral nerve grafts implantation into the rat hippocampal dentate gyrus. METHODS: Experiment was carried out on female rats divided into three experimental groups: NO - non-ovariectomized, OV - ovariectomized and E - heterogeneous group with various 17-beta-estradiol substitution after ovariectomy. Proliferating cells were labeled with BrdU. Brains were subjected to immunohistochemical procedures to visualize nestin, GFAP and estrogen receptors (ERalpha and ERbeta). RESULTS: Proliferation rate was highest in E groups with estrogen levels resembling that in proestrus phase. Ovariectomy resulted in higher than in NO group number of new neurons, while high hyperestrogenemia worsened the results. The proportions of nestin-labeled cells correlated in similar way with different hormonal state. We found also distinct co-localization of nestin and GFAP in E group (proestrus). It may suggest the presence of radial glia, a potential source of new neurons in adult mammals. Nerve graft induced ERalpha expression at the site of injury in all groups. Distribution of ERbeta in hippocampus was estradiol-dose-dependent and correlated with cell proliferation. CONCLUSION: In our model, 17-beta-estradiol and predegenerated nerve graft implantation had synergistic effect on hippocampal neurogenesis. PMID- 19675527 TI - Deletion of the Met receptor in the collecting duct decreases renal repair following ureteral obstruction. AB - Hepatocyte growth factor and its receptor, Met, activate biological pathways necessary for repair and regeneration following kidney injury. The Met receptor is expressed in multiple cell types within the kidney, each of which is capable of regulating fibrotic responses. To specifically address the role of the Met receptor in the adult collecting duct during renal injury, a conditional knockout mouse (Met(fl/fl);HoxB7-Cre) was generated and tested using unilateral ureteral obstruction, a model of nephron injury, fibrosis, and repair. Following obstruction in these mice there was increased expression of collagens I and IV along with plasminogen activator inhibitor 1, a known regulator of matrix degradation, compared to ureteral obstructed non-flox littermates. There were trends toward increased interstitial fibrosis, infiltration of the interstitium, and acute tubular necrosis in the knockout mice despite similar degrees of hydronephrosis to the control littermates. The Met(fl/fl);HoxB7-Cre mice; however, had reduced tubular cell proliferation and kidney regenerative capacity after release of the obstruction, thus leading to diminished functional recovery. We suggest that Met receptor signaling in the collecting duct acts as a major regulator of cell survival and propagation of the repair process with a possible secondary role to diminish inflammatory and fibrotic responses. PMID- 19675528 TI - Non-infected hemodialysis catheters are associated with increased inflammation compared to arteriovenous fistulas. AB - Although hemodialysis catheters predispose to infection which, in turn, causes inflammation, we studied whether they induce inflammation independent of infection. We compared the level of the inflammatory marker C-reactive protein (CRP) in maintenance hemodialysis patients, comparing those dialyzed using a non infected catheter to those using arteriovenous fistulas. All incident patients had catheters and fistula placement at dialysis initiation. In 35 patients the fistulas matured, the catheters were removed and the patients were evaluated at 6 months (catheter-fistula). These results were compared to 15 patients in whom the fistula did not mature and catheter use persisted for 6 months (catheter catheter). There was a significant 82% reduction in the CRP level in the catheter fistula group but a 16% increase in the catheter-catheter group at 6 months. The changes in CRP did not differ by gender, diabetes status, or by race, and was not correlated with a change in phosphorus, age, or urea reduction ratio at 1 month following hemodialysis initiation. Decreased CRP was associated with increased hemoglobin and albumin. Patients with persistent fistula use from dialysis initiation through 6 months had consistently low CRP levels over that time period. Our study shows that catheters might contribute to increased inflammation independent of infection, and supports avoidance of catheters and a timely conversion to fistulas with catheter removal. PMID- 19675529 TI - Health related quality of life and the CKD patient: challenges for the nephrology community. AB - The compromised health-related quality of life (HRQOL) of patients with chronic kidney disease is now well documented. The recent mandate by the Center for Medicare Services in the United States that all dialysis units monitor HRQOL as a condition of coverage has focused attention on the importance of these measures. The challenge for the nephrology care team is understanding how to interpret and utilize the information obtained from these HRQOL measurements. Can HRQOL of these patients be improved? The present review addresses this issue by commenting on strategies that have been used to improve the HRQOL of chronic kidney disease patients. A systematic approach is suggested for nephrology care providers to attempt to evaluate and improve the HRQOL of CKD patients. PMID- 19675530 TI - Acute phosphate nephropathy. AB - Acute phosphate nephropathy (APhN) is a clinical pathological entity characterized by acute and subsequent chronic renal failure following exposure to oral sodium phosphate (OSP) bowel purgatives. Renal biopsy findings include acute and chronic tubular injury with prominent tubular and interstitial calcium phosphate deposits. Risk factors for APhN include older age, female gender, hypertension, chronic kidney disease (CKD), and treatment with angiotensin converting enzyme inhibitors, angiotensin receptor blockers, and diuretics. The pathomechanism of APhN involves hypovolemia-induced avid proximal salt and water reabsorption, delivery of a large phosphate load to the distal nephron, and precipitation of calcium phosphate in the distal tubule and collecting duct. To date, 37 cases of biopsy-proven APhN have been reported, and epidemiologic studies have produced inconsistent results regarding the incidence of acute kidney injury (AKI) following the use of OSP purgatives. OSP solution was withdrawn from the market in December of 2008, but OSP tablets, offered by prescription only, remain available. Prevention of APhN is best achieved by avoiding OSP in high-risk patients, aggressive hydration before, during, and after OSP administration, minimizing the dose of OSP, and maintaining a minimum of a 12 h interval between OSP administrations. PMID- 19675532 TI - Anti-brain autoantibodies and altered excitatory neurotransmitters in obsessive compulsive disorder. AB - Although serum autoantibodies directed against basal ganglia (BG) implicate autoimmunity in the pathogenesis of obsessive-compulsive disorder (OCD), it is unclear whether these antibodies can cross the blood-brain barrier to bind against BG or other components of the OCD circuit. It is also unclear how they might lead to hyperactivity in the OCD circuit. We examined this by investigating the presence of autoantibodies directed against the BG or thalamus in the serum as well as CSF of 23 OCD patients compared with 23 matched psychiatrically normal controls using western blot. We further investigated CSF amino acid (glutamate, GABA, taurine, and glycine) levels and also examined the extent to which these levels were related to the presence of autoantibodies. There was evidence of significantly more binding of CSF autoantibodies to homogenate of BG as well as to homogenate of thalamus among OCD patients compared with controls. There was no significant difference in binding between patient and control sera except for a trend toward more bands to BG and thalamic protein corresponding to 43 kD among OCD patients compared with controls. CSF glutamate and glycine levels were also significantly higher in OCD patients compared with controls, and further multivariate analysis of variance showed that CSF glycine levels were higher in those OCD patients who had autoantibodies compared with those without. The results of our study implicate autoimmune mechanisms in the pathogenesis of OCD and also provide preliminary evidence that autoantibodies against BG and thalamus may cause OCD by modulating excitatory neurotransmission. PMID- 19675531 TI - Dopamine D1 receptor (DRD1) genetic polymorphism: pleiotropic effects on heritable renal traits. AB - Because dopamine D(1) receptors (DRD1) influence renal sodium transport and vascular hemodynamics, we examined whether genetic polymorphisms play a role in renal function. We conducted polymorphism discovery across the DRD1 open reading frame and its 5'-UTR and then performed association studies with estimated glomerular filtration rate (eGFR), plasma creatinine (pCr), and fractional excretion of uric acid (FeUA). We used a twin/family group of 428 subjects from 195 families and a replication cohort of 677 patients from the Kaiser health-care organization sampled from the lower percentiles of diastolic blood pressures. Although the coding region lacked common non-synonymous variants, we identified two polymorphisms in the DRD1 5'-UTR (G-94A, A-48G) that occurred with frequencies of 15 and 30%, respectively. In the twin/family study, renal traits were highly heritable, such that DRD1 G-94A significantly associated with eGFR, pCr, and FeUA. Homozygotes for the G-94A minor allele (A/A) exhibited lower eGFR, higher pCr, and lower FeUA. No effects were noted for DRD1 A-48G. Patients in the Kaiser group had similar effects of G-94A on eGFR and pCr. Kidney cells transfected with the -94A variant but not the wild type vectors had increased receptor density. Because the -94A allele is common and may reduce glomerular capillary hydrostatic pressure, G-94A profiling may aid in predicting survival of renal function in patients with progressive renal disease. PMID- 19675533 TI - Differential effects of tri-allelic 5-HTTLPR polymorphisms in healthy subjects on mood and stress performance after tryptophan challenge. AB - Earlier data suggest that a polymorphism at the serotonin (5-HT) transporter gene (5-HTTLPR) may affect depression particularly in the face of stress due to interactions between 5-HT vulnerability and stress exposure. However, this interaction between 5-HT transporter-linked transcriptional promoter region (5 HTTLPR), 5-HT vulnerability and the affective effects of stress exposure has not yet been investigated. As participants with short-allele 5-HTTLPR genotypes may exhibit enhanced 5-HT vulnerability, this study examines the effects of tryptophan challenge on stress reactivity and performance in healthy participants with S'/S' vs L'/L' genotypes. Sixteen healthy subjects with homozygotic short alleles (S'/S'=S/L(G,) L(G)/L(G)) and 14 subjects with homozygotic long alleles (L'/L'=L(A)/L(A)) of the 5-HTTLPR were tested in a double-blind placebo controlled design under acute stress exposure following tryptophan challenge or placebo. Although there were no 5-HTTLPR-related differences in stress responses, significant beneficial effects of tryptophan challenge on mood and stress performance were exclusively found in participants with S'/S' genotypes. These findings suggest greater brain 5-HT vulnerability to tryptophan manipulations in participants with S'/S' as compared with L'/L' 5-HTTLPR genotypes. This apparent genetic 5-HT vulnerability may become a meaningful risk factor for depression when brain 5-HT falls below functional need in the face of real severe stressful life events. PMID- 19675535 TI - Modulation of limbic circuitry predicts treatment response to antipsychotic medication: a functional imaging study in schizophrenia. AB - The regional neuronal changes taking place in the early and late stages of antipsychotic treatment are still not well characterized in humans. In addition, it is not known whether these regional changes are predictive of or are correlated with treatment response. Using PET with 15O, we evaluated the time course of regional cerebral blood flow (rCBF) patterns generated by a first (haloperidol) and a second (olanzapine) generation antipsychotic drug in patients with schizophrenia during a 6-week treatment trial. Patients were initially scanned after withdrawal of all psychotropic medication (2 weeks), and then blindly randomized to treatment with haloperidol (n=12) or olanzapine (n=17) for a period of 6 weeks. Patients were scanned again after 1 and 6 weeks of treatment. All assessments, including scanning sessions, were obtained in a double-blind manner. As hypothesized, we observed rCBF changes that were common to both the drugs, implicating cortico-subcortical and limbic neuronal networks in antipsychotic action. In addition, in these regions, some patterns seen at weeks 1 and 6 were distinctive, indexing neuronal changes related to an early (ventral striatum, hippocampus) and consolidated (anterior cingulate/medial frontal cortex) stage of drug response. Finally, both after 1 and 6 weeks of treatment, we observed differential patterns of rCBF activation between good and poor responders. After 1 week of treatment, greater rCBF increase in the ventral striatum and greater decrease in the hippocampus were associated with good response. PMID- 19675536 TI - Repeated homotypic stress elevates 2-arachidonoylglycerol levels and enhances short-term endocannabinoid signaling at inhibitory synapses in basolateral amygdala. AB - Psychosocial stress is a risk factor for development and exacerbation of neuropsychiatric illness. Repeated stress causes biochemical adaptations in endocannabinoid (eCB) signaling that contribute to stress-response habituation, however, the synaptic correlates of these adaptations have not been examined. Here, we show that the synthetic enzyme for the eCB 2-arachidonoylglycerol (2 AG), diacylglycerol (DAG) lipase alpha, is heterogeneously expressed in the amygdala, and that levels of 2-AG and precursor DAGs are increased in the basolateral amygdala (BLA) after 10 days, but not 1 day, of restraint stress. In contrast, arachidonic acid was decreased after both 1 and 10 days of restraint stress. To examine the synaptic correlates of these alterations in 2-AG metabolism, we used whole-cell electrophysiology to determine the effects of restraint stress on depolarization-induced suppression of inhibition (DSI) in the BLA. A single restraint stress exposure did not alter DSI compared with control mice. However, after 10 days of restraint stress, DSI duration, but not magnitude, was significantly prolonged. Inhibition of 2-AG degradation with MAFP also prolonged DSI duration; the effects of repeated restraint stress and MAFP were mutually occlusive. These data indicate that exposure to repeated, but not acute, stress produces neuroadaptations that confer BLA neurons with an enhanced capacity to elevate 2-AG content and engage in 2-AG-mediated short-term retrograde synaptic signaling. We suggest stress-induced enhancement of eCB mediated suppression of inhibitory transmission in the BLA could contribute to affective dysregulation associated with chronic stress. PMID- 19675539 TI - Pharmacological discrimination of extinction and reconsolidation of contextual fear memory by a potentiator of AMPA receptors. AB - Conditioned fear memory, once formed through fear conditioning, is modulated by reexposure of individuals to a conditioned stimulus. The reexposure reactivates the fear memory, which induces reconsolidation of the memory first, and then extinction of the fear response. Both attenuating the former and facilitating the latter are effective in reducing the fear response, and these findings are potentially translatable to the enhancement of exposure therapy for complex anxiety disorders. Currently, there is no drug that is established to modulate either reconsolidation or extinction selectively, which are thought to be independent processes. Here, we report that an extinction-facilitating AMPA potentiator, 4-[2-(phenylsulfonylamino)ethylthio]-2,6-difluoro-phenoxyacetamide (PEPA), does not act on the reconsolidation of fear memory formed by contextual fear conditioning in mice. The freezing rates observed in contextually conditioned mice following short reexposure (3 min) to the context were not influenced by intraperitoneal or intra-amygdala administration of PEPA. The same short reexposure to the context enhanced freezing responses in mice that were similarly administered D-cycloserine (DCS), a drug that facilitates both extinction and reconsolidation, and this enhancement of freezing responses in mice intraperitoneally administered DCS was abolished by propranolol, a drug that suppresses reconsolidation. At the same doses used in the short reexposure experiments, PEPA and DCS facilitated extinction of the fear response induced by long reexposure to the context and suppressed reinstatement of the conditioned fear memory. PEPA and DCS did not affect reextinction. These results suggest that PEPA acts on extinction of contextual fear memory without having detectable influences on its reconsolidation. PMID- 19675540 TI - Pharmacokinetic studies in hemodialysis patients. AB - Intermittent hemodialysis is the primary supportive therapy for patients with end stage renal disease, who commonly receive several drugs to treat both their underlying disease and the conditions that arise during long-term hemodialysis therapy. Many of these drugs are dialyzable, and their hemodialytic removal may compromise therapeutic efficacy if appropriate supplementary doses are not given. Emergency hemodialysis may also be life-saving for patients who have received drug overdoses or have ingested toxic substances. Optimal therapy in both these clinical settings is critically dependent on the availability of reliable information from well-designed pharmacokinetic studies. PMID- 19675538 TI - Impairments of probabilistic response reversal and passive avoidance following catecholamine depletion. AB - Catecholamines, particularly dopamine, have been implicated in various aspects of the reward function including the ability to learn through reinforcement and to modify flexibly responses to changing reinforcement contingencies. We examined the impact of catecholamine depletion (CD) achieved by oral administration of alpha-methyl-paratyrosine (AMPT) on probabilistic reversal learning and passive avoidance (PA) in 15 female subjects with major depressive disorder in full remission (RMDD) and 12 healthy female controls. The CD did not affect significantly the acquisition phase of the reversal learning task. However, CD selectively impaired reversal of the 80-20 contingency pair. In the PA learning task, CD was associated with reduced responding toward rewarding stimuli, although the RMDD and control subjects did not differ regarding these CD-induced changes in reward processing. Interestingly, the performance decrement produced by AMPT on both of these tasks was associated with the level of decreased metabolism in the perigenual anterior cingulate cortex. In an additional examination using the affective Stroop task we found evidence for impaired executive attention as a trait abnormality in MDD. In conclusion, this study showed specific effects of CD on the processing of reward-related stimuli in humans and confirms earlier investigations that show impairments of executive attention as a neuropsychological trait in affective illness. PMID- 19675537 TI - Stress-induced potentiation of cocaine reward: a role for CRF R1 and CREB. AB - Both clinical and preclinical research have shown that stress can potentiate drug use; however, the underlying mechanisms of this interaction are unknown. Previously, we have shown that a single exposure to forced swim (FS) reinstates extinguished conditioned place preference (CPP) to cocaine and that cAMP response element binding protein (CREB) is necessary for this response. CREB can be activated by corticotropin releasing factor (CRF) receptor type 1 (CRF(R1)) binding, which mediates neuroendocrine and behavioral responses to stress as well as to drugs of abuse. The present experiments investigate whether changes in cocaine reward elicited by previous exposure to stress are mediated by CREB and/or CRF(R1). Chronic exposure to FS in advance of conditioning enhances cocaine CPP in wild-type mice, but this is blocked in CREB-deficient mice. In addition, pretreatment with the CRF(R1) antagonist, antalarmin, before FS exposure blocks this stress-induced enhancement of cocaine CPP. Furthermore, FS induced increase in phosphorylated CREB (pCREB), specifically in the lateral septum (LS) and nucleus accumbens (NAc) is also blocked by antalarmin. Taken together, these studies suggest that both CREB and CRF(R1) activation are necessary for stress-induced potentiation of drug reward. PMID- 19675542 TI - Challenges in analgesic drug development. PMID- 19675541 TI - Pharmacokinetic study of pegylated liposomal CKD-602 (S-CKD602) in patients with advanced malignancies. AB - S-CKD602 is a pegylated liposomal formulation of CKD-602. This study is the first to evaluate the factors affecting the high interpatient variability in the pharmacokinetic disposition of S-CKD602. S-CKD602 was administered intravenously (i.v.) every 3 weeks as part of a phase I study. Pharmacokinetics studies of the liposomal encapsulated and released CKD-602 in plasma were performed. The pharmacokinetic variability of S-CKD602 is associated with both linear and nonlinear clearances. Patients > or =60 years of age have a 2.7-fold higher exposure of S-CKD602 as compared with patients <60 years of age (P = 0.02). Patients with a lean body composition have a higher plasma exposure of S-CKD602 (P = 0.02). Patients who have received prior therapy with pegylated liposomal doxorubicin (PLD) have a 2.2-fold higher exposure of S-CKD602 as compared with patients who have not received PLD (P = 0.045). Prolonged exposure of the encapsulated drug in plasma over 1-2 weeks provides significant pharmacologic advantages. The high interpatient variability in the pharmacokinetic disposition of S-CKD602 was associated with age, body composition, saturable clearance, and prior PLD therapy. PMID- 19675543 TI - From trial and error to trial simulation. Part 2: an appraisal of current beliefs in the design and analysis of clinical trials for antidepressant drugs. AB - Study design factors are partly to blame for the high failure rate in trials with antidepressant drugs. Clinical trial simulation (CTS) allows the investigation of the influence of design characteristics on important aspects of clinical trials such as power and type I error. Using CTS scenarios, we evaluated the impact of population size, randomization ratio, frequency of assessments, dropout mechanisms, clinical end point, and statistical method on the outcome of clinical trials with antidepressant drugs. The results reveal that (i) an increase in the frequency of visits does not increase statistical power, (ii) a skewed randomization for a placebo or comparator arm may decrease statistical power, and (iii) analysis of the percentage of responders should be avoided. CTS should become best practice in the optimization of study design. To date, no other statistical approach has enabled such comprehensive evaluation of the factors contributing to study failure in depression. PMID- 19675534 TI - Cortico-Basal Ganglia reward network: microcircuitry. AB - Many of the brain's reward systems converge on the nucleus accumbens, a region richly innervated by excitatory, inhibitory, and modulatory afferents representing the circuitry necessary for selecting adaptive motivated behaviors. The ventral subiculum of the hippocampus provides contextual and spatial information, the basolateral amygdala conveys affective influence, and the prefrontal cortex provides an integrative impact on goal-directed behavior. The balance of these afferents is under the modulatory influence of dopamine neurons in the ventral tegmental area. This midbrain region receives its own complex mix of excitatory and inhibitory inputs, some of which have only recently been identified. Such afferent regulation positions the dopamine system to bias goal directed behavior based on internal drives and environmental contingencies. Conditions that result in reward promote phasic dopamine release, which serves to maintain ongoing behavior by selectively potentiating ventral subicular drive to the accumbens. Behaviors that fail to produce an expected reward decrease dopamine transmission, which favors prefrontal cortical-driven switching to new behavioral strategies. As such, the limbic reward system is designed to optimize action plans for maximizing reward outcomes. This system can be commandeered by drugs of abuse or psychiatric disorders, resulting in inappropriate behaviors that sustain failed reward strategies. A fuller appreciation of the circuitry interconnecting the nucleus accumbens and ventral tegmental area should serve to advance discovery of new treatment options for these conditions. PMID- 19675544 TI - Preface. PMID- 19675545 TI - Possible involvement of basement membrane damage in skin photoaging. AB - Aging of sun-exposed skin is accelerated by three major environmental factors: UV radiation, dryness, and oxidation. UV radiation exposure is the most influential factor in skin aging (so-called photoaging). To find ways to protect against damage caused by UV exposure and to delay photoaging, we studied internal changes of sun-exposed skin compared with those of sun-protected skin. We found that the basement membrane (BM) at the dermal-epidermal junction (DEJ) of sun-exposed skin becomes damaged and multilayered and partly disrupted compared with that of sun protected skin. BM plays important roles in maintaining a healthy epidermis and dermis, and repeated damage destabilizes the skin, accelerating the aging process. Matrix metalloproteinases (MMPs) and urinary plasminogen activator are increased in UV-irradiated skin. MMPs are detected in the cornified layer in sun exposed skin, but not in sun-protected skin. Using skin-equivalent models, we found that MMPs and plasmin cause BM damage and that the reconstruction of BM is enhanced by inhibiting these proteinases, as well as by increasing the synthesis of BM components. Enhancement of BM repair mechanisms may be a useful strategy in retarding photoaging.Journal of Investigative Dermatology Symposium Proceedings (2009) 14, 2-7; doi:10.1038/jidsymp.2009.5. PMID- 19675546 TI - Progeroid syndromes and UV-induced oxidative DNA damage. AB - Progeroid syndromes are a group of diseases characterized by signs of premature aging. These syndromes comprise diseases such as Werner syndrome, Bloom syndrome, Rothmund-Thomson syndrome, Hutchinson-Gilford syndrome, Fanconi anemia, and ataxia-telangiectasia, as well as xeroderma pigmentosum, trichothiodystrophy, and Cockayne syndrome. Clinical symptoms of premature aging are skin atrophy with loss of cutaneous elasticity, dysfunction of cutaneous appendices, degeneration of the central nervous system and an increased susceptibility for malignant tumors. Genetic defects in the repair of DNA damage can lead to progeroid syndromes, and it is becoming increasingly evident that direct DNA damage and indirect damage by highly reactive oxygen species play central roles in aging. The clinical signs of progeroid syndromes and the molecular aspects of UV (ultraviolet radiation)-induced oxidative stress in aging are discussed.Journal of Investigative Dermatology Symposium Proceedings (2009) 14, 8-14; doi:10.1038/jidsymp.2009.6. PMID- 19675547 TI - Effects of infrared radiation and heat on human skin aging in vivo. AB - Sunlight damages human skin, resulting in a wrinkled appearance. Since natural sunlight is polychromatic, its ultimate effects on the human skin are the result of not only the action of each wavelength separately, but also interactions among the many wavelengths, including UV, visible light, and infrared (IR). In direct sunlight, the temperature of human skin rises to about 40 degrees C following the conversion of absorbed IR into heat. So far, our knowledge of the effects of IR radiation or heat on skin aging is limited. Recent work demonstrates that IR and heat exposure each induces cutaneous angiogenesis and inflammatory cellular infiltration, disrupts the dermal extracellular matrix by inducing matrix metalloproteinases, and alters dermal structural proteins, thereby adding to premature skin aging. This review provides a summary of current research on the effects of IR radiation and heat on aging in human skin in vivo.Journal of Investigative Dermatology Symposium Proceedings (2009) 14, 15-19; doi:10.1038/jidsymp.2009.7. PMID- 19675548 TI - Matrix-degrading metalloproteinases in photoaging. AB - UV radiation from the sun impacts skin health adversely through complex, multiple molecular pathways. Premature skin aging (photoaging) is among the most widely appreciated harmful effects of chronic exposure to solar UV radiation. Extensive damage to the dermal connective tissue is a hallmark of photoaged skin. Disruption of the normal architecture of skin connective tissue impairs skin function and causes it to look aged. UV irradiation induces expression of certain members of the matrix metalloproteinase (MMP) family, which degrade collagen and other extracellular matrix proteins that comprise the dermal connective tissue. Although the critical role of MMPs in photoaging is undeniable, important questions remain. This article summarizes our current understanding of the role of MMPs in the photoaging process and presents new data that (1) describe the expression and regulation by UV irradiation of all members of the MMP family in human skin in vivo and (2) quantify the relative contributions of epidermis and dermis to the expression of UV irradiation-induced MMPs in human skin in vivo.Journal of Investigative Dermatology Symposium Proceedings (2009) 14, 20-24; doi:10.1038/jidsymp.2009.8. PMID- 19675549 TI - Telomere-mediated effects on melanogenesis and skin aging. AB - UV-induced melanogenesis (tanning) and "premature aging" or photoaging result in large part from DNA damage. This article reviews data tying both phenomena to telomere-based DNA damage signaling and develops a conceptual framework in which both responses may be understood as cancer-avoidance protective mechanisms.Journal of Investigative Dermatology Symposium Proceedings (2009) 14, 25-31; doi:10.1038/jidsymp.2009.9. PMID- 19675551 TI - Mechanism of UVB-induced wrinkling of the skin: paracrine cytokine linkage between keratinocytes and fibroblasts leading to the stimulation of elastase. AB - In clinical studies, the formation of facial wrinkles has been closely linked to the loss of elastic properties of the skin. Repetitive irradiation of animal skin with UVB radiation at suberythemal doses significantly reduces its elastic properties, resulting in the formation of wrinkles. Repetitive UVB irradiation elicits a marked alteration in the three-dimensional structure of elastic fibers, which is closely associated with a subsequent reduction in the elastic properties of the skin. Although UVB irradiation stimulates the activity of fibroblast elastases in the dermis, a synthetic inhibitor specific for fibroblast elastases prevents wrinkle formation. The close interrelationships among wrinkle formation, elastic properties, and elastic fiber linearity are revealed by the effects of different concentrations of the elastase inhibitor (R(2)>0.9), suggesting that enhanced elastase activity by dermal fibroblasts plays a pivotal role in the UVB wrinkling mechanism. In in vitro studies we identified a paracrine linkage between keratinocytes and fibroblasts that leads to wrinkle formation through the upregulation of fibroblast elastases. These studies support our hypothesis for a mechanism of wrinkle formation by which cytokine expression is activated in epidermal keratinocytes by UVB radiation and triggers dermal fibroblasts to increase their expression of elastase.Journal of Investigative Dermatology Symposium Proceedings (2009) 14, 36-43; doi:10.1038/jidsymp.2009.11. PMID- 19675550 TI - Short- and long-term effects of UV radiation on the pigmentation of human skin. AB - The incidence of skin cancer, including cutaneous melanoma, has risen substantially in recent years, and epidemiological and laboratory studies show that UV radiation is a major causative factor of this increase. UV damage also underlies photoaging of the skin, and these deleterious effects of UV can be, in part, prevented in skin with higher levels of constitutive pigmentation. We review the clinical studies we have made in recent years regarding the rapid and the long-term responses of the pigmentary system in human skin to UV exposure.Journal of Investigative Dermatology Symposium Proceedings (2009) 14, 32 35; doi:10.1038/jidsymp.2009.10. PMID- 19675552 TI - Role of mitochondria in photoaging of human skin: the defective powerhouse model. AB - The exact pathogenesis of photoaging of the skin is not yet known. Earlier, a number of molecular pathways explaining one or more characteristics of photoaged skin have been described, but a unifying mechanistic concept is still missing. Here we propose the "Defective Powerhouse Model of Premature Skin Aging", which reconciles most of the earlier conducted research as one concept. In this model, the persistence of UV radiation-induced mtDNA deletions or the infrared radiation induced disturbance of the electron flow of the mitochondrial electron transport chain leads to inadequate energy production in dermal fibroblasts. As a consequence of this defective powerhouse, retrograde mitochondrial signaling pathways are triggered that then they transduce functional and structural alterations in the skin. This model, which is supported by a growing number of recent studies, is of direct clinical importance in preventing and treating photoaging in human skin.Journal of Investigative Dermatology Symposium Proceedings (2009) 14, 44-49; doi:10.1038/jidsymp.2009.1. PMID- 19675553 TI - Reactive oxygen species in HaCaT keratinocytes after UVB irradiation are triggered by intracellular Ca(2+) levels. AB - It is recognized that reactive oxygen species (ROS) are responsible for skin damage due to UVB-radiation (UVB-R). However, the triggering substance(s) for ROS generation after UVB-R is uncertain with respect to the activation of NADPH oxidase (Nox), xanthine oxidase (XOD), and respiratory chain-chain reactions in mitochondria. As a first step in identifying the trigger(s) for UVB-induced ROS generation, we examined the relationship between Ca(2+) levels and ROS generation in HaCaT keratinocytes. UVB-R exposure of HaCaT keratinocytes resulted in an immediate elevation of ROS that recurred 7 hours later. This was accompanied by immediately elevated intracellular Ca(2+) . A Ca(2+) chelating agent, BAPTA, abolished the elevation of ROS after UVB-R completely. In addition, exogenous H(2)O(2) did not increase intracellular Ca(2+) levels. This suggests that intracellular Ca(2+) is the first trigger for UVB-induced ROS generation.Journal of Investigative Dermatology Symposium Proceedings (2009) 14, 50-52; doi:10.1038/jidsymp.2009.12. PMID- 19675554 TI - Molecular basis of tobacco smoke-induced premature skin aging. AB - Although it is now widely recognized that tobacco smoke has negative effects on the skin, the molecular mechanisms underlying its skin-aging effects remain uncertain. Epidemiological studies indicate that tobacco smoking is a strong independent predictor of facial wrinkle formation and other aspects of premature skin aging. Recent in vivo studies in humans and mice provided the first direct evidence that tobacco smoke causes premature skin aging, and they have begun to reveal the molecular changes in the skin that occur in response to it. Water soluble tobacco smoke extract, which predominantly produces oxidative stress when applied topically to cultured skin fibroblasts, impairs collagen biosynthesis. Matrix metalloproteinases, which degrade collagen, are induced dose-dependently by tobacco smoke extract as well as by other constituents that trigger the aryl hydrocarbon receptor (AhR), a ligand-dependent transcription factor that mediates the toxicity of several environmental contaminants, including photoproducts in the body generated by UVB radiation. Tobacco smoke also contains many non-water soluble constituents that activate the AhR pathway. Our most recent studies using hexane-soluble tobacco extract indicate that activation of the AhR pathway may play a role in the premature skin-aging effects of tobacco smoke exposure.Journal of Investigative Dermatology Symposium Proceedings (2009) 14, 53-55; doi:10.1038/jidsymp.2009.13. PMID- 19675555 TI - Non-sunscreen photoprotection: antioxidants add value to a sunscreen. AB - The association between ultraviolet radiation (UVR) exposure and both skin cancer and photo-aging is well documented. In addition to the conventional organic chemical and physical-mineral type sunscreens, other non-sunscreen protective strategies have been developed. These include topically applied botanical extracts and other antioxidants as well as topical DNA repair enzymes. Standard terms of photoprotection such as sun protection factor (SPF) do not accurately reflect the photoprotection benefits of these materials. For example, in spite of minimal SPF, tea extract containing polyphenols such as (-)-epigallocatechin-3 gallate (EGCG) has been shown to protect against UV-induced DNA damage and immune suppression, in part through its ability to reduce oxidative stress and inhibit NF-kB. The addition of botanical antioxidants and vitamins C and E to a broad spectrum sunscreen may further decrease UV-induced damage compared with sunscreen alone. These agents have been shown to enhance protection against UV-induced epidermal thickening, overexpression of MMP-1and MMP-9, and depletion of CD1a(+) Langerhans cells. Non-sunscreen materials such as botanical extracts, antioxidants, and DNA repair enzymes can contribute value when applied topically to human skin in vivo.Journal of Investigative Dermatology Symposium Proceedings (2009) 14, 56-59; doi:10.1038/jidsymp.2009.14. PMID- 19675556 TI - Involvement of a forkhead transcription factor, FOXO1A, in UV-induced changes of collagen metabolism. AB - Transcription factors belonging to the forkhead box gene, group O (FOXOs) family have been found to be crucial in downstream suppression of life-shortening effects of the insulin/insulin-like growth factor-1 (IGF-1) signaling pathway, which accelerates aging by suppressing FOXOs. Thus, FOXOs could hold the key for counteracting aging. Although FOXOs may play a critical role in aging, the effects of FOXOs on UV-induced changes of collagen metabolism by dermal fibroblasts are unknown. In this study, UV-induced changes in FOXO1a expression and the roles of FOXO1a in the regulation of collagen synthesis and matrix metalloproteinase (MMPs) expression in human dermal fibroblasts were investigated. In UVA- or UVB-irradiated fibroblasts, the expression of FOXO1A mRNA decreased significantly. The expression of type I collagen (COLIAI) also decreased. On the other hand, MMP-1 and MMP-2 mRNA levels increased. FOXO1A-small interfering RNA transfection induced the downregulation of FOXO1A expression, it also induced a decrease in COLIAI expression, and it increased MMP-1 and MMP-2 expression. These changes are similar to those observed in UV-irradiated fibroblasts. Furthermore, FOXO1a-peptide induced opposite changes in COLIAI, MMP 1, and MMP-2 expression. Therefore, FOXO1a is involved in the UV-induced changes of type I collagen and MMPs expression.Journal of Investigative Dermatology Symposium Proceedings (2009) 14, 60-62; doi:10.1038/jidsymp.2009.2. PMID- 19675557 TI - DNA repair and cytokine responses. AB - As sunscreens do not provide complete protection against solar/UV radiation, alternative protective strategies are necessary to cope with the increasing incidence of skin cancer. These strategies include the reduction of UVR-induced DNA damage by the topical application of bacterial DNA repair enzymes. Recent evidence suggests that nucleotide excision repair, the physiological repair system that is mostly responsible for the removal of UVR-mediated DNA damage, can be modulated by cytokines, including IL-12, IL-18, and alpha-melanocyte stimulating hormone. The mechanisms involved and the biological as well as the potential therapeutic implications of these findings are discussed.Journal of Investigative Dermatology Symposium Proceedings (2009) 14, 63-66; doi:10.1038/jidsymp.2009.3. PMID- 19675558 TI - The pathogenesis of photoaging: the role of neutrophils and neutrophil-derived enzymes. AB - The hallmark of photoaged skin is solar elastosis, which is probably an end product of elastic fiber degradation. Exposure of human skin to a certain threshold of UV, infrared radiation (IR), and heat leads to an influx of neutrophils. These neutrophils are packed with potent proteolytic enzymes capable of degrading collagen and, particularly, elastic fibers. Neutrophil-derived proteolytic enzymes are held responsible for the extracellular matrix (ECM) damage observed in several non-dermatological conditions. Furthermore, neutrophil elastase, a major product of neutrophils, is strongly associated with solar elastosis in mice. Taken together with our data that show in vivo proteolytic activity of neutrophil-derived elastase and matrix metalloproteinases (MMPs) in UV-exposed skin, we have hypothesized earlier that neutrophils are major contributors to the photoaging process. Although several groups have shown that MMPs are also induced in skin exposed to relatively low doses of UV, IR, and heat, clinical data indicate that high(er) doses of UV, IR, and heat are necessary to induce photoaging or photoaging-like pathology in the skin. Therefore, we propose that MMPs generated by suberythemogenic doses of UV and low doses of IR/heat are involved in cellular processes other than ECM degradation.Journal of Investigative Dermatology Symposium Proceedings (2009) 14, 67-72; doi:10.1038/jidsymp.2009.15. PMID- 19675559 TI - Regulation of skin pigmentation and thickness by Dickkopf 1 (DKK1). AB - Dickkopf 1 (DKK1), an inhibitor of Wnt signaling, not only functions as a head inducer during development, but also regulates joint remodeling and bone formation, which suggests roles for DKK1 in the pathogenesis of rheumatoid arthritis and multiple myeloma. We recently demonstrated that levels of DKK1 in palmoplantar dermal fibroblasts are physiologically higher than those observed in non-palmoplantar dermal fibroblasts. Thus, the DKK1-rich mesenchyme in palmoplantar dermis affects the overlying epithelium and induces a palmoplantar phenotype in the epidermis. More specifically, DKK1 suppresses melanocyte function and growth through the regulation of microphthalmia-associated transcription factor (MITF) and beta-catenin. Furthermore, DKK1 induces the expression of keratin 9 and alpha-Kelch-like ECT2-interacting protein (alphaKLEIP) but downregulates the expression of beta-catenin, glycogen synthase kinase 3beta, protein kinase C, and proteinase-activated receptor-2 (PAR-2) in keratinocytes. Treatment of reconstructed skin with DKK1 reproduces the hypopigmentation and thickening of skin through Wnt/beta-catenin signaling. These studies elucidate why human palmoplantar skin is thicker and paler than non palmoplantar skin through the secretion of DKK1 by fibroblasts that affect the overlying epidermis. Thus, DKK1 may be useful for reducing skin pigmentation and for thickening photo-aged skin and palmoplantar wounds caused by diabetes mellitus and rheumatic skin diseases.Journal of Investigative Dermatology Symposium Proceedings (2009) 14, 73-75; doi:10.1038/jidsymp.2009.4. PMID- 19675560 TI - Direct visualization of mouse brain oxygen distribution by electron paramagnetic resonance imaging: application to focal cerebral ischemia. AB - Electron paramagnetic resonance imaging (EPRI) is a new modality for visualizing O(2) distribution in tissues, such as the brain after stroke or after administration of drugs of abuse. We have recently shown that 3 acetoxymethoxycarbonyl-2,2,5,5-tetramethyl-1-pyrrolidinyloxyl [1] is a pro imaging agent that can cross the blood-brain barrier. After hydrolysis by esterases, the anion of 3-carboxy-2,2,5,5-tetramethyl-1-tetramethyl-1 pyrrolidinyloxyl [2] is trapped in brain tissue. In this study, we investigated the feasibility of using this to map the changes of O(2) concentration in mouse brain after focal ischemia. The decrease in tissue O(2) concentration in the ischemic region of mouse brain was clearly visualized by EPRI. The hypoxic zone mapped by EPRI was spatially well correlated with the infarction area in the brain imaged by diffusion-weighted magnetic resonance imaging (MRI). Finally, we observed a decrease in the size of the hypoxic region when the mouse breathed higher levels of O(2). This finding suggests that EPRI with specifically designed nitroxides is a promising imaging modality for visualizing O(2) distribution in brain tissue, especially in an ischemic brain. We believe that this imaging method can be used for monitoring the effects of therapeutic intervention aimed at enhancing brain O(2) supply, which is crucial in minimizing brain injury after stroke. PMID- 19675561 TI - Dissociation between vascular endothelial growth factor receptor-2 and blood vessel density in the caudate nucleus after chronic hydrocephalus. AB - Chronic hydrocephalus (CH) is characterized by the presence of ventricular enlargement, decreased cerebral blood flow (CBF), and brain tissue oxygen delivery. Although the underlying pathophysiological role of vascular endothelial growth factor (VEGF) is not clear, ischemic-hypoxic events in CH are known to trigger its release. Previously, we have shown increased VEGF receptor-2 (VEGFR 2) and blood vessel density (BVd) in the hippocampus after CH. We investigated changes in neuronal and glial VEGFR-2 density and BVd in the caudate nucleus in an experimental model of CH. Animals with CH were divided into short term (ST, 2 to 4 weeks) and long term (LT, 12 to 16 weeks) and were compared with surgical controls (SCs, 12 to 16 weeks). The cellular and BVds were estimated using immunohistochemical and stereological counting methods. Overall, percentage (%)VEGFR-2 neurons were approximately two times greater in CH (ST, LT) than in SC. By comparison, glial cell %VEGFR-2 was greater by 10% to 17% in ST and 4% to 11% lower in LT compared with that in SC. Blood vessel density was significantly lower in CH than in SC in the superficial caudate. Changes in cerebrospinal fluid ventricular volume and pressure, as well as in CBF did not correlate with either VEGFR-2 or BVd. These observed findings suggest that destructive forces may outweigh angiogenic forces and possibly show a disassociation between VEGFR-2 and BV expressions. PMID- 19675562 TI - Total cerebral blood flow and hippocampal volume in patients with arterial disease. The SMART-MR study. AB - It has been suggested that compared with other brain tissues, the hippocampus in particular is vulnerable to chronic hypoperfusion. We investigated whether total parenchymal cerebral blood flow (pCBF) was associated with hippocampal atrophy, and also whether this relationship was modified by white matter lesions (WMLs). In a cross-sectional analysis within the SMART-MR (Second Manifestations of ARTerial disease-magnetic resonance) study, which is a cohort study among patients with arterial disease, total CBF (tCBF) and hippocampal volume were assessed in 392 patients (mean age: 62+/-9 years, 84% men). Total CBF was expressed in per 100 mL brain volume for obtaining pCBF. Manual volumetric measurements of the hippocampus were carried out on a three-dimensional fast field-echo T1-weighted magnetic resonance imaging scan with isotropic voxels. Automated brain segmentation was used to quantify volumes of the WML and the total brain. A linear regression analysis showed that reduced pCBF was not associated with smaller hippocampal volume after adjustments were made for age and sex. The association attenuated further after additional adjustments were made for vascular risk factors, lacunar infarcts, and WMLs (beta=0.01 mL per s.d. decrease in pCBF; 95% confidence interval: -0.06 to 0.08). The association was not modified by WML (P-value for interaction term pCBF*WML=0.84). We found no evidence of the fact that lower parenchymal blood flow contributes to the neurodegeneration of the hippocampus in a population of patients with arterial disease. PMID- 19675563 TI - Increased cerebral blood volume and oxygen consumption in neonatal brain injury. AB - With the increasing interest in treatments for neonatal brain injury, bedside methods for detecting and assessing injury status and evolution are needed. We aimed to determine whether cerebral tissue oxygenation (StO(2)), cerebral blood volume (CBV), and estimates of relative cerebral oxygen consumption (rCMRO(2)) determined by bedside frequency-domain near-infrared spectroscopy (FD-NIRS) have the potential to distinguish neonates with brain injury from those with non-brain issues and healthy controls. We recruited 43 neonates < or =15 days old and >33 weeks gestational age (GA): 14 with imaging evidence of brain injury, 29 without suspicion of brain injury (4 unstable, 6 stable, and 19 healthy). A multivariate analysis of variance with Newman-Keuls post hoc comparisons confirmed group similarity for GA and age at measurement. StO(2) was significantly higher in brain injured compared with unstable neonates, but not statistically different from stable or healthy neonates. Brain-injured neonates were distinguished from all others by significant increases in CBV and rCMRO(2). In conclusion, although NIRS measures of StO(2) alone may be insensitive to evolving brain injury, increased CBV and rCMRO(2) seem to be useful for detecting neonatal brain injury and suggest increased neuronal activity and metabolism occurs acutely in evolving brain injury. PMID- 19675564 TI - Phosphatidylserine-containing liposomes promote maximal survival of retinal neurons after ischemic injury. AB - We investigated the systemic effect of liposomes bearing apoptotic signals on the level of inflammation and neuronal death induced by ischemia-reperfusion (IR). Using a model of retinal ischemia, we showed that treatment with phosphatidylserine (PS) and phosphatidylcholine (PC) liposomes significantly reduced the expression of proinflammatory genes, including that of Il1b, Il6, Ccl2, Ccl5, Cxcl10, and Icam1, 24 h after reperfusion. Phosphatidylserine liposome treatment was the most efficient and correlated with significantly reduced neuronal death in the retina 7 days after reperfusion. The results of our study indicate that therapeutic strategy based on mimicking a systemic increase in apoptotic signaling can significantly reduce central nervous system damage induced by IR and improve neurologic outcome. PMID- 19675565 TI - Neuroprotective role of lactate after cerebral ischemia. AB - It is well established that lactate can be used as an energy substrate by the brain by conversion to pyruvate and a subsequent oxidation in the mitochondria. Knowing the need for readily metabolizable substrates directly after ischemia and the protective effect of lactate after excitotoxicity, the aim of this study was to investigate whether lactate administration directly after ischemia could be neuroprotective. In vitro, the addition of 4 mmol/L L-lactate to the medium of rat organotypic hippocampal slices, directly after oxygen and glucose deprivation (OGD), protected against neuronal death, whereas a higher dose of 20 mmol/L was toxic. In vivo, after middle cerebral artery occlusion in the mouse, an intracerebroventricular injection of 2 microL of 100 mmol/L L-lactate, immediately after reperfusion, led to a significant decrease in lesion size, which was more pronounced in the striatum, and an improvement in neurologic outcome. A later injection 1 h after reperfusion did not reduce lesion size, but significantly improved neurologic outcome, which is an important point in the context of a potential clinical application. Therefore, a moderate increase in lactate after ischemia may be a therapeutic tool. PMID- 19675567 TI - The structural basis of tail-anchored membrane protein recognition by Get3. AB - Targeting of newly synthesized membrane proteins to the endoplasmic reticulum is an essential cellular process. Most membrane proteins are recognized and targeted co-translationally by the signal recognition particle. However, nearly 5% of membrane proteins are 'tail-anchored' by a single carboxy-terminal transmembrane domain that cannot access the co-translational pathway. Instead, tail-anchored proteins are targeted post-translationally by a conserved ATPase termed Get3. The mechanistic basis for tail-anchored protein recognition or targeting by Get3 is not known. Here we present crystal structures of yeast Get3 in 'open' (nucleotide free) and 'closed' (ADP.AlF(4)(-)-bound) dimer states. In the closed state, the dimer interface of Get3 contains an enormous hydrophobic groove implicated by mutational analyses in tail-anchored protein binding. In the open state, Get3 undergoes a striking rearrangement that disrupts the groove and shields its hydrophobic surfaces. These data provide a molecular mechanism for nucleotide regulated binding and release of tail-anchored proteins during their membrane targeting by Get3. PMID- 19675568 TI - The importance of niches for the maintenance of species diversity. AB - Ecological communities characteristically contain a wide diversity of species with important functional, economic and aesthetic value. Ecologists have long questioned how this diversity is maintained. Classic theory shows that stable coexistence requires competitors to differ in their niches; this has motivated numerous investigations of ecological differences presumed to maintain diversity. That niche differences are key to coexistence, however, has recently been challenged by the neutral theory of biodiversity, which explains coexistence with the equivalence of competitors. The ensuing controversy has motivated calls for a better understanding of the collective importance of niche differences for the diversity observed in ecological communities. Here we integrate theory and experimentation to show that niche differences collectively stabilize the dynamics of experimental communities of serpentine annual plants. We used field parameterized population models to develop a null expectation for community dynamics without the stabilizing effects of niche differences. The population growth rates predicted by this null model varied by several orders of magnitude between species, which is sufficient for rapid competitive exclusion. Moreover, after two generations of community change in the field, Shannon diversity was over 50 per cent greater in communities stabilized by niche differences relative to those exhibiting dynamics predicted by the null model. Finally, in an experiment manipulating species' relative abundances, population growth rates increased when species became rare--the demographic signature of niche differences. Our work thus provides strong evidence that species differences have a critical role in stabilizing species diversity. PMID- 19675570 TI - Satellite-based estimates of groundwater depletion in India. AB - Groundwater is a primary source of fresh water in many parts of the world. Some regions are becoming overly dependent on it, consuming groundwater faster than it is naturally replenished and causing water tables to decline unremittingly. Indirect evidence suggests that this is the case in northwest India, but there has been no regional assessment of the rate of groundwater depletion. Here we use terrestrial water storage-change observations from the NASA Gravity Recovery and Climate Experiment satellites and simulated soil-water variations from a data integrating hydrological modelling system to show that groundwater is being depleted at a mean rate of 4.0 +/- 1.0 cm yr(-1) equivalent height of water (17.7 +/- 4.5 km(3) yr(-1)) over the Indian states of Rajasthan, Punjab and Haryana (including Delhi). During our study period of August 2002 to October 2008, groundwater depletion was equivalent to a net loss of 109 km(3) of water, which is double the capacity of India's largest surface-water reservoir. Annual rainfall was close to normal throughout the period and we demonstrate that the other terrestrial water storage components (soil moisture, surface waters, snow, glaciers and biomass) did not contribute significantly to the observed decline in total water levels. Although our observational record is brief, the available evidence suggests that unsustainable consumption of groundwater for irrigation and other anthropogenic uses is likely to be the cause. If measures are not taken soon to ensure sustainable groundwater usage, the consequences for the 114,000,000 residents of the region may include a reduction of agricultural output and shortages of potable water, leading to extensive socioeconomic stresses. PMID- 19675569 TI - Direct activation of protein kinases by unanchored polyubiquitin chains. AB - TRAF6 is a ubiquitin ligase that is essential for the activation of NF-kappaB and MAP kinases in several signalling pathways, including those emanating from the interleukin 1 and Toll-like receptors. TRAF6 functions together with a ubiquitin conjugating enzyme complex consisting of UBC13 (also known as UBE2N) and UEV1A (UBE2V1) to catalyse Lys 63-linked polyubiquitination, which activates the TAK1 (also known as MAP3K7) kinase complex. TAK1 in turn phosphorylates and activates IkappaB kinase (IKK), leading to the activation of NF-kappaB. Although several proteins are known to be polyubiquitinated in the IL1R and Toll-like receptor pathways, it is not clear whether ubiquitination of any of these proteins is important for TAK1 or IKK activation. By reconstituting TAK1 activation in vitro using purified proteins, here we show that free Lys 63 polyubiquitin chains, which are not conjugated to any target protein, directly activate TAK1 by binding to the ubiquitin receptor TAB2 (also known as MAP3K7IP2). This binding leads to autophosphorylation and activation of TAK1. Furthermore, we found that unanchored polyubiquitin chains synthesized by TRAF6 and UBCH5C (also known as UBE2D3) activate the IKK complex. Disassembly of the polyubiquitin chains by deubiquitination enzymes prevented TAK1 and IKK activation. These results indicate that unanchored polyubiquitin chains directly activate TAK1 and IKK, suggesting a new mechanism of protein kinase regulation. PMID- 19675571 TI - To crash or not to crash? Probability in the management of benign lacrimal gland tumours. PMID- 19675572 TI - Multifocal electroretinography changes in patients on ethambutol therapy. AB - PURPOSE: To evaluate the multifocal electroretinography (mfERG) changes in patients on ethambutol therapy. METHODS: A cross-sectional observational study of 17 visually asymptomatic patients receiving antituberculosis therapy with ethambutol. Patients underwent complete ophthalmic examination and mfERG recordings. The first-order mfERG N1 and P1 response amplitudes and implicit times of six concentric rings were analysed and compared with 17 normal age similar controls. Correlation analyses were performed between the patients' mfERG parameters with parameters of ethambutol usage (daily dose of ethambutol per body weight, duration of ethambutol therapy, cumulative dose of ethambutol, and cumulative dose of ethambutol per body weight). RESULTS: The mean duration of ethambutol therapy was 3.6 months (range: 2-9 months) and the mean daily dose per body weight was 13.2 mg/kg/day. Analysis of response amplitude measures showed no significant difference in the mfERG N1 and P1 response amplitudes between the ethambutol and control groups at all ring eccentricities (P>0.05). For implicit times, there were significant delays in the mfERG P1 implicit times of rings 4-6 in the ethambutol group compared with controls (P=0.012 to P=0.041). Correlation analyses showed no significant correlation between the mfERG and ethambutol parameters (P>0.05). CONCLUSIONS: The mfERG findings suggested that visually asymptomatic patients receiving ethambutol therapy might have localized mild electrophysiological changes involving the peripheral macula. These changes might be related to localized alteration of metabolism or physiological changes associated with ethambutol therapy. PMID- 19675574 TI - Serum thrombopoietin level and thrombocytopenia during the neonatal period in infants with Down's syndrome. AB - BACKGROUND: The pathogenesis of thrombocytopenia during the neonatal period in Down's syndrome (DS) infants remains unclear. OBJECTIVE: To elucidate kinetic changes of serum thrombopoietin (TPO) level and platelet count, and their correlation in DS neonates. STUDY DESIGN: Twelve DS infants (male/female: 7/5, term/late preterm: 10/2) born between 1997 and 2007 were included. Blood samples were serially collected during the neonatal period and serum TPO levels were determined in 44 sera using an enzyme-linked immunosorbent assay. RESULTS: Thrombocytopenia <150 x 10(9) per liter was observed in seven (58%) patients. In 12 DS patients, the median TPO value showed 2.86 fmol ml(-1) on day 0, rose to 4.64 fmol ml(-1) on day 2, and thereafter decreased to 4.30 fmol ml(-1) on day 5, 2.40 fmol ml(-1) on days 11-15, and 1.75 fmol ml(-1) on days 28-30. This kinetics parallels that in historical non-DS controls. In 35 pair sample analysis from 11 patients without transient myeloproliferative disease, TPO level inversely correlated with platelet count (r=-0.38, P=0.023). However, there was no significant difference in TPO concentrations between thrombocytopenic and non thrombocytopenic DS individuals. CONCLUSIONS: This is the first study to describe the relationship between TPO level and platelet count in neonates with DS. Median TPO levels and their kinetic changes in DS neonates are comparable to those in non-DS controls. In contrast to earlier findings in several studies showing higher TPO concentrations in thrombocytopenic non-DS newborns than those in non thrombocytopenic counterparts, the response of the TPO system to thrombocytopenia in DS during the neonatal period seems suboptimal. PMID- 19675576 TI - Triterpenes promote keratinocyte differentiation in vitro, ex vivo and in vivo: a role for the transient receptor potential canonical (subtype) 6. AB - It has been shown recently that triterpenes inhibit cancer cell growth of various cell types in vitro. In this work, the effect of highly purified triterpenes (TE) with betulin as the major compound (>80% w/w) on cell proliferation, apoptosis, and differentiation of human keratinocytes was analyzed in vitro, ex vivo, and in vivo. In vitro, TE increased calcium influx into primary keratinocytes and upregulated various differentiation markers including keratin 10. TE also specifically increased the expression of the non-selective transient receptor potential canonical (subtype) 6 (TRPC6) in keratinocytes, and knocking down TRPC6 inhibited keratin 10 upregulation. Ex vivo, in human skin explants TE induced the expression of TRPC6 in the epidermis and increased DNA fragmentation of terminally differentiating keratinocytes. Topical treatment with TE of actinic keratoses, that represent in situ squamous cell carcinomas with disturbed epithelial differentiation, resulted in downgrading of aberrant Ki67 expression and upregulation of keratin 10 in vivo. Our data indicate that TE promotes keratinocyte differentiation in vitro and in vivo. This effect seems to be mediated at least in part by TRPC6. PMID- 19675575 TI - Skin-resident T cells: the ups and downs of on site immunity. AB - The cutaneous surface of a normal adult individual contains approximately 20 billion T cells, nearly twice the number present in the entire circulation. Recent studies have shown a role for these cells in both normal immunity and in inflammatory skin diseases such as psoriasis. Regulatory T cells protect against autoimmune reactions to self antigens and assist in the resolution of cutaneous inflammation. However, they can also shield tumors from immune detection, allow latent infections to persist and can dysfunction under the conditions present in inflammatory skin diseases. Th17 T cells protect organisms against extracellular pathogens but also have a key role in the pathogenesis of psoriasis. Evidence suggests that effector memory T cells produced during immune reactions survive and persist long term within the skin, providing local and rapid protection against pathogen reexposure. This review summarizes the current understanding of how skin-resident T cells contribute to normal and aberrant immunity in the skin. PMID- 19675577 TI - PPARdelta activation promotes stratum corneum formation and epidermal permeability barrier development during late gestation. AB - The goal of epidermal ontogenesis is to form a stratum corneum (SC), which is required for post-natal permeability barrier function. The regulation of epidermal ontogenesis is poorly understood, but nuclear hormone receptors have been shown to have an important function. As peroxisome proliferator-activated receptor-delta (PPARdelta) is very abundant in fetal epidermis and PPARdelta activation stimulates differentiation and permeability barrier formation in adults, we hypothesized that PPARdelta might regulate epidermal ontogenesis. Treatment of fetal rat explants with the PPARdelta ligand, GW 610742X, accelerates permeability barrier development, evidenced by a decrease in transepidermal water loss and an enhanced outside-in barrier function, attributable to the presence of more mature lamellar membranes in the SC and enhanced expression of loricrin and involucrin. Similarly, the intra-amniotic administration of GW 610742X also accelerates the formation of the SC and permeability barrier development. Finally, in PPARdelta-deficient mice the formation of the SC and the expression of differentiation-related proteins were delayed on days 16.5 and 17.5 of gestation. However, at later stages (day 18.5 and after birth), there were no differences between wild-type- and PPARdelta deficient mice, indicating only a transient delay in epidermal ontogenesis. These studies show that PPARdelta has a role in SC formation and permeability barrier development. PMID- 19675578 TI - The small heat-shock protein Hsp27 undergoes ERK-dependent phosphorylation and redistribution to the cytoskeleton in response to dual leucine zipper-bearing kinase expression. AB - Hsp27, a small heat-shock protein, has important roles in many cellular processes, including cytoskeleton dynamics, cell differentiation, and apoptosis. Its expression in normal epidermis correlates with differentiation; however, little is known about the regulatory mechanisms involved. In this study, we report that Hsp27 undergoes upregulation, phosphorylation, and redistribution to the cytoskeleton during the late phase of epidermal keratinocyte differentiation. Our results also show that the expression of the dual leucine zipper-bearing kinase (DLK), an upstream activator of the MAP kinase pathways, is sufficient by itself to induce Hsp27 phosphorylation, cell periphery localization, and redistribution to the insoluble protein fraction (cytoskeleton) in poorly differentiated keratinocytes. This redistribution correlates with the insolubilization of cornified envelope-associated proteins such as involucrin. Interestingly, the effects of DLK on Hsp27 were blocked by PD98059, a selective inhibitor of the extracellular signal-regulated protein kinase (ERK) pathway. Moreover, downregulation of Hsp27 by small interfering RNA in epithelial cells expressing DLK was accompanied by attenuated expression of involucrin in the cytoskeleton. Thus, these observations suggest that the DLK-ERK signaling pathway may act as a regulator of the interaction that occurs between Hsp27 and the cytoskeleton during the formation of the cornified cell envelope, a process conferring to the skin its crucial barrier function. PMID- 19675580 TI - Blue-light irradiation regulates proliferation and differentiation in human skin cells. AB - Sunlight influences the physiology of the human skin in beneficial as well as harmful ways, as has been shown for UV light. However, little is known about the effects of other wavelengths of solar irradiation. In this study we irradiated human keratinocytes and skin-derived endothelial cells with light-emitting-diode devices of distinct wavelengths to study the effects on cell physiology. We found that light at wavelengths of 632-940 nm has no effect, but irradiation with blue light at 412-426 nm exerts toxic effects at high fluences. Light at 453 nm is nontoxic up to a fluence of 500 J/cm(2). At nontoxic fluences, blue light reduces proliferation dose dependently by up to 50%, which is attributable to differentiation induction as shown by an increase of differentiation markers. Experiments with BSA demonstrate that blue-light irradiation up to 453 nm photolytically generates nitric oxide (NO) from nitrosated proteins, which is known to initiate differentiation in skin cells. Our data provide evidence for a molecular mechanism by which blue light may be effective in treating hyperproliferative skin conditions by reducing proliferation due to the induction of differentiation. We observed a photolytic release of NO from nitrosated proteins, indicating that they are light acceptors and signal transducers up to a wavelength of 453 nm. PMID- 19675579 TI - Regeneration of multilineage skin epithelia by differentiated keratinocytes. AB - Although homeostasis of rapidly renewing tissues like skin epithelia is maintained by stem cells, the committed progeny of stem cells in the basal layer of epidermis retain regenerative potential and are capable of forming epidermis in response to environmental cues. It is not clear, however, at what point within the epidermal lineage keratinocytes lose this regenerative potential. In this study, we examined the extent of tissue formation by post-mitotic differentiated keratinocytes. We show that cultures of mouse keratinocytes that were, by all measures, differentiated were able to reform a self-renewing, hair-bearing skin when transplanted onto suitable sites in vivo. Genetic labeling and lineage tracing studies in combination with an involucrin-driven Cre/lox reporter system confirmed that transplanted differentiated keratinocytes were indeed the source of the regenerated skin. More importantly, analysis of early stages of skin regeneration showed hallmarks of dedifferentiation of transplanted differentiated keratinocytes. These data indicate that commitment to differentiation does not prohibit cells from re-entering the cell cycle, de-differentiating, and acquiring "stemness". These findings suggest that epidermis can use different strategies for homeostasis and tissue regeneration. PMID- 19675581 TI - Multiple loci comprising immune-related genes regulate experimental neuroinflammation. AB - A 58 Mb region on rat chromosome 4 known to regulate experimental autoimmune encephalomyelitis (EAE) was genetically dissected. High-resolution linkage analysis in an advanced intercross line (AIL) revealed four quantitative trait loci (QTLs), Eae24-Eae27. Both Eae24 and Eae25 regulated susceptibility and severity phenotypes, whereas Eae26 regulated severity and Eae27 regulated susceptibility. Analyses of the humoral immune response revealed that the levels of serum anti-myelin oligodendrocyte glycoprotein (MOG) immunoglobin G1 (IgG1) antibodies are linked to Eae24 and anti-MOG IgG2b antibodies are linked to both Eae24 and Eae26. We tested the parental DA strain and six recombinant congenic strains that include overlapping fragments of this region in MOG-EAE. Eae24 and Eae25 showed significant protection during the acute phase of EAE, whereas Eae25 and Eae26 significantly modified severity but not susceptibility. The smallest congenic fragment, which carries Eae25 alone, influenced both susceptibility and severity, and protected from the chronic phase of disease. These results support the multiple QTLs identified in the AIL. By demonstrating several QTLs comprising immune-related genes, which potentially interact, we provide a significant step toward elucidation of the polygenically regulated pathogenesis of MOG-EAE and possibly multiple sclerosis (MS), and opportunities for comparative genetics and testing in MS case-control cohorts. PMID- 19675583 TI - A population genetics study of the familial Mediterranean fever gene: evidence of balancing selection under an overdominance regime. AB - Familial Mediterranean Fever (FMF) is a recessively inherited systemic autoinflammatory disease caused by mutations in the MEFV gene. The frequency of different disease alleles is extremely high in multiple populations from the Mediterranean region, suggesting heterozygote advantage. Here, we characterize the sequence variation and haplotype structure of the MEFV 3' gene region (from exon 5 to the 3' UTR) in seven human populations. In non-African populations, we observed high levels of nucleotide variation, an excess of intermediate-frequency alleles, reduced population differentiation and a genealogy with two common haplotypes separated by deep branches. These features are suggestive of balancing selection having acted on this region to maintain one or more selected alleles. In line with this finding, an excess of heterozygotes was observed in Europeans and Asians, suggesting an overdominance regime. Our data, together with the earlier demonstration that the MEFV exon 10 has been subjected to episodic positive selection over primate evolution, provide evidence for an adaptive role of nucleotide variation in this gene region. Our data suggest that further studies aimed at clarifying the role of MEFV variants might benefit from the integration of molecular evolutionary and functional analyses. PMID- 19675582 TI - A functional SNP in the regulatory region of the decay-accelerating factor gene associates with extraocular muscle pareses in myasthenia gravis. AB - Complement activation in myasthenia gravis (MG) may damage muscle endplate and complement regulatory proteins such as decay-accelerating factor (DAF) or CD55 may be protective. We hypothesize that the increased prevalence of severe extraocular muscle (EOM) dysfunction among African MG subjects reported earlier may result from altered DAF expression. To test this hypothesis, we screened the DAF gene sequences relevant to the classical complement pathway and found an association between myasthenics with EOM paresis and the DAF regulatory region c. 198C>G SNP (odds ratio=8.6; P=0.0003). This single nucleotide polymorphism (SNP) results in a twofold activation of a DAF 5'-flanking region luciferase reporter transfected into three different cell lines. Direct matching of the surrounding SNP sequence within the DAF regulatory region with the known transcription factor binding sites suggests a loss of an Sp1-binding site. This was supported by the observation that the c.-198C>G SNP did not show the normal lipopolysaccharide induced DAF transcriptional upregulation in lymphoblasts from four patients. Our findings suggest that at critical periods during autoimmune MG, this SNP may result in inadequate DAF upregulation with consequent complement-mediated EOM damage. Susceptible individuals may benefit from anti-complement therapy in addition to immunosuppression. PMID- 19675584 TI - Progress and Prospects: genetic treatments for disorders of bones and joints. AB - Gene therapies directed toward the treatment of arthritis and tissue repair continue to be the most active areas of research for bone and joint diseases. In the past 2 years, two trials in rheumatoid arthritis have been completed. a Phase I study reporting safety and a Phase I/II study that has yet to be published. An additional, small study has reported the first evidence of clinical efficacy. Two Phase I trials of gene therapy for osteoarthritis have also been initiated. There is much preclinical activity in developing AAV vectors for future trials in the gene therapy of arthritis. Research into tissue repair and regeneration remains at the preclinical stage, but a considerable volume of research attests to the promise of gene transfer in this arena, especially in the context of bone healing. For tissue repair, the major research questions are still which genes to use and how best to deliver them. PMID- 19675586 TI - Angiogenesis and hypertension: an update. AB - The purpose of this review is to provide a basic understanding of the important relationship between microvascular remodelling, angiogenesis and hypertension, that is, provide an overview of recent experimental and clinical evidence from anti-hypertensive and pro- and anti-angiogenic therapy with respect to hypertension and microvascular structure. Microvascular rarefaction, that is, a loss of terminal arterioles and capillaries, is found in most forms of human and experimental arterial hypertension. This further increases peripheral resistance, and aggravates hypertension and hypertension-induced target organ damage. In some cases with a genetic predisposition, hypertension is preceded by a loss of microvessels. Therefore, new therapies aimed at reversing microvascular rarefaction potentially represent candidate treatments of hypertension. The microvasculature is formed by the continuous balance between de novo angiogenesis and microvascular regression. Imbalanced angiogenesis, in addition to functional shut-off of blood flow, contributes to microvascular rarefaction. Numerous clinical trials assessing anti-angiogenic agents in cancer patients show that this therapy leads to microvascular rarefaction and causes or aggravates hypertension. The development of specific pro-angiogenic treatment to correct hypertension or ischaemic disorders, however, it is still in its infancy. On the other hand, long-term treatment by classic anti-hypertensive therapies that present vasodilator activity can correct for hypertension-associated rarefaction in man. PMID- 19675587 TI - Relationship between stem cell factor/c-kit expression in peripheral blood and blood pressure. AB - There is a high prevalence of hypertension and hypertension-related vascular disease in humans. Studies show that the expression of stem cell factor (SCF)/c kit signalling proteins is relatively high during blood vessel repair. The aim of this study was to investigate the relationship between blood pressure (BP) and the expression of SCF/c-kit in peripheral blood. We carried out a cross-sectional analysis of 141 subjects in the health examination centre of our hospital. Information including waist circumference, BP, plasma glucose and serum lipids for each subject was collected. Endothelin-1 (ET-1) and tumour necrosis factor alpha (TNF-alpha) levels in peripheral blood were determined by radio-immunity assay. Expression levels of SCF and its receptor, c-kit, in peripheral blood were measured by enzyme-linked immunosorbent assay (ELISA). We found a positive correlation between plasma SCF/c-kit levels and BP in these patients (SCF: hypertension 907.1+/-52.3 vs pre-hypertension 834.6+/-47.6 vs normal control 768.8+/-44.1 ng l(-1); c-kit: hypertension 13.2+/-1.6 vs pre-hypertension 11.1+/ 2.1 vs normal control 9.6+/-1.5 mg l(-1), P<0.01). SCF/c-kit levels were also positively correlated with ET-1 and TNF-alpha levels (ET-1: hypertension 155.5+/ 12.1 vs pre-hypertension 142.0+/-11.2 vs normal control 117.9+/-10.3 ng l(-1); TNF-alpha: hypertension 14.7+/-3.9 vs pre-hypertension 11.6+/-4.2 vs normal control 8.1+/-2.8 ng l(-1), P<0.01). Multiple linear regression analyses showed that SCF, c-kit and ET-1 are independent predictors for systolic blood pressure, and that SCF, c-kit, ET-1 and TNF-alpha are independent predictors for diastolic blood pressure. SCF/c-kit levels increase with BP levels are positively correlated with ET-1 and TNF-alpha levels. PMID- 19675588 TI - Glomerular filtration rate is related to dipping pattern in ambulatory blood pressure monitoring--a cross-sectional population-based study. AB - A non-dipping pattern in ambulatory blood pressure monitoring (ABPM) increases the risk of cardiovascular disease. The association between renal function and the dipping pattern has not been studied in a random middle-aged population. This is a cross-sectional population-based study of 226 males and 234 females aged 40 to 62 years. Renal function was assessed with estimated glomerular filtration rate (eGFR). Non-dipping status was defined as a reduction of <10% between the daytime and the nighttime systolic BP. Non-dippers represented 18.7% of the study population. Their mean eGFR was 79.1 (s.d. 15.7) ml min(-1) per 1.73 m(2) as compared with a mean eGFR of 84.1 (s.d. 16.2) ml min(-1) per 1.73 m(2) in dippers (P=0.005); this difference remained significant after adjustments. Subjects in the lowest and in the middle eGFR tertiles had an independently increased risk of non-dipping in comparison with those in the highest eGFR tertile (odd ratios (OR), 2.34 (95% confidence interval (CI), 1.18 to 4.63) and OR, 2.01 (95% CI, 1.06 to 3.83), respectively). This study showed that even a minor deterioration in renal function is associated with increased risk of non-dipping pattern in ABPM in a random middle-aged population. PMID- 19675589 TI - Multitargeted tyrosine kinase inhibitor stimulates expression of IL-6 and activates JAK2/STAT5 signaling in acute myelogenous leukemia cells. PMID- 19675592 TI - Immunology and Cell Biology's impact factor continues to rise. PMID- 19675593 TI - Highly efficient eradication of intracranial glioblastoma using Eg5 siRNA combined with HVJ envelope. AB - Hemagglutinating virus of Japan envelope (HVJ-E) vector with inactivated replication-defective Sendai virus was originally developed as a versatile drug delivery system. Recently, we have shown the direct tumor-killing activity of HVJ E itself without therapeutic molecules in prostate cancer cells. Although human glioblastoma cells were also sensitive to HVJ-E treatment, complete eradication was not achieved using HVJ-E alone. Here, to develop more effective therapeutic strategy of glioblastoma, we enhanced the anti-tumor activity by incorporating Short interfering RNA (siRNA) of mitotic motor protein Eg5 into HVJ-E. Treatment with HVJ-E-containing Eg5 siRNA induced monopolar spindle formation and resulted in cell-cycle arrest and apoptosis. When HVJ-E-containing Eg5 siRNA was directly injected into an intradermal tumor xenograft, all mice became tumor-free. Similar results were observed in the intracranial tumor xenografts. The survival time of treated mice was significantly prolonged when HVJ-E was used. Histological examination showed that tumors remained in the brain after treatment with HVJ-E containing negative control siRNA, but no tumors were detected after treatment with HVJ-E-containing Eg5 siRNA. This remarkable anti-tumor response was likely due to a synergistic effect of Eg5 siRNA and HVJ-E. Thus, this combination shows promise as an attractive novel therapy for glioblastoma. PMID- 19675594 TI - Adenoviral-mediated gene transfer restores plasma ADAMTS13 antigen and activity in ADAMTS13 knockout mice. AB - ADAMTS13 is a plasma metalloprotease that regulates the size of the von Willebrand factor (VWF) multimers. Genetic or acquired deficiency of ADAMTS13 causes thrombotic thrombocytopenic purpura (TTP) in humans. Plasma infusion is the treatment of choice for patients with congenital ADAMTS13 deficiency. However, this practice exposes patients to the risk of infections, allergies and fluid volume overload. The search for alternative treatments is required. Here, we tested the ability of systemically administered adenovirus encoding human ADAMTS13 to restore the deficient protein in the circulation of Adamts13(-/-) mice. Injection of the adenovirus efficiently transduced the liver, kidney, lung, heart and spleen, resulting in the secretion of ADAMTS13 into plasma. A reduced area of thrombi was observed when blood from Ad-ADAMTS13-treated mice was perfused over a collagen-coated surface in a parallel plate flow chamber compared with blood of Ad-betaGal-treated controls. The secreted ADAMTS13 protein was functionally active even after 2 months from injection. The data provide the proof of principle for developing a novel therapy for the correction of ADAMTS13 deficiency in patients with hereditary TTP. PMID- 19675599 TI - Lessons from Antarctica. PMID- 19675595 TI - Linking phylogenetic and functional diversity to nutrient spiraling in microbial mats from Lower Kane Cave (USA). AB - Microbial mats in sulfidic cave streams offer unique opportunities to study redox based biogeochemical nutrient cycles. Previous work from Lower Kane Cave, Wyoming, USA, focused on the aerobic portion of microbial mats, dominated by putative chemolithoautotrophic, sulfur-oxidizing groups within the Epsilonproteobacteria and Gammaproteobacteria. To evaluate nutrient cycling and turnover within the whole mat system, a multidisciplinary strategy was used to characterize the anaerobic portion of the mats, including application of the full cycle rRNA approach, the most probable number method, and geochemical and isotopic analyses. Seventeen major taxonomic bacterial groups and one archaeal group were retrieved from the anaerobic portions of the mats, dominated by Deltaproteobacteria and uncultured members of the Chloroflexi phylum. A nutrient spiraling model was applied to evaluate upstream to downstream changes in microbial diversity based on carbon and sulfur nutrient concentrations. Variability in dissolved sulfide concentrations was attributed to changes in the abundance of sulfide-oxidizing microbial groups and shifts in the occurrence and abundance of sulfate-reducing microbes. Gradients in carbon and sulfur isotopic composition indicated that released and recycled byproduct compounds from upstream microbial activities were incorporated by downstream communities. On the basis of the type of available chemical energy, the variability of nutrient species in a spiraling model may explain observed differences in microbial taxonomic affiliations and metabolic functions, thereby spatially linking microbial diversity to nutrient spiraling in the cave stream ecosystem. PMID- 19675600 TI - A change of tone. PMID- 19675609 TI - Journal club. An evolutionary biologist learns how to be remembered: cheat someone. PMID- 19675611 TI - LHC hopes for collisions by Christmas. PMID- 19675612 TI - Science advisers mull priorities. PMID- 19675613 TI - Flu database rocked by legal row. PMID- 19675614 TI - Ice-core researchers hope to chill out. PMID- 19675615 TI - Climate data spat intensifies. PMID- 19675616 TI - Return of the rat. PMID- 19675617 TI - Satellite data show Indian water stocks shrinking. PMID- 19675624 TI - Atmospheric science: Fixing the sky. PMID- 19675625 TI - Ape and human similarities can be deceptive. PMID- 19675626 TI - Speed of reporting isn't the issue when your work is scooped. PMID- 19675627 TI - European bodies can help to tackle TB worldwide. PMID- 19675632 TI - Mathematical physics: A tight squeeze. PMID- 19675633 TI - Stem cells: Escaping fates with open states. PMID- 19675634 TI - Ecology: Elementary factors. PMID- 19675635 TI - Cancer: More than kin and less than kind. PMID- 19675636 TI - Chemical biology: Protein picker. PMID- 19675639 TI - Technology: Hydrogen-fuelled vehicles. PMID- 19675638 TI - Microbial genetics: Love the one you're with. PMID- 19675640 TI - Metalloproteins. PMID- 19675641 TI - Structure-function relationships of anaerobic gas-processing metalloenzymes. AB - Reactions involving H(2), N(2), CO, CO(2) and CH(4) are likely to have been central to the origin of life. This is indicated by the active-site structures of the enzymes involved, which are often reminiscent of minerals. Through the combined efforts of protein crystallography, various types of spectroscopy, theoretical calculations and model chemistry, it has been possible to put forward plausible mechanisms for gas-based metabolism by extant microorganisms. Although the reactions are based on metal centres, the protein matrix regulates reactivity and substrate and product trafficking through internal pathways, specific ligation and dielectricity. PMID- 19675642 TI - Metalloproteins and metal sensing. AB - Almost half of all enzymes must associate with a particular metal to function. An ambition is to understand why each metal-protein partnership arose and how it is maintained. Metal availability provides part of the explanation, and has changed over geological time and varies between habitats but is held within vital limits in cells. Such homeostasis needs metal sensors, and there is an ongoing search to discover the metal-sensing mechanisms. For metalloproteins to acquire the right metals, metal sensors must correctly distinguish between the inorganic elements. PMID- 19675643 TI - Function and biogenesis of iron-sulphur proteins. AB - Iron-sulphur (Fe-S) clusters have long been recognized as essential and versatile cofactors of proteins involved in catalysis, electron transport and sensing of ambient conditions. Despite the relative simplicity of Fe-S clusters in terms of structure and composition, their synthesis and assembly into apoproteins is a highly complex and coordinated process in living cells. Different biogenesis machineries in both bacteria and eukaryotes have been discovered that assist Fe-S protein maturation according to uniform biosynthetic principles. The importance of Fe-S proteins for life is documented by an increasing number of diseases linked to these components and their biogenesis. PMID- 19675644 TI - Molybdenum cofactors, enzymes and pathways. AB - The trace element molybdenum is essential for nearly all organisms and forms the catalytic centre of a large variety of enzymes such as nitrogenase, nitrate reductases, sulphite oxidase and xanthine oxidoreductases. Nature has developed two scaffolds holding molybdenum in place, the iron-molybdenum cofactor and pterin-based molybdenum cofactors. Despite the different structures and functions of molybdenum-dependent enzymes, there are important similarities, which we highlight here. The biosynthetic pathways leading to both types of cofactor have common mechanistic aspects relating to scaffold formation, metal activation and cofactor insertion into apoenzymes, and have served as an evolutionary 'toolbox' to mediate additional cellular functions in eukaryotic metabolism. PMID- 19675645 TI - Mechanistic considerations of halogenating enzymes. AB - In nature, halogenation is a strategy used to increase the biological activity of secondary metabolites, compounds that are often effective as drugs. However, halides are not particularly reactive unless they are activated, typically by oxidation. The pace of discovery of new enzymes for halogenation is increasing, revealing new metalloenzymes, flavoenzymes, S-adenosyl-L-methionine (SAM) dependent enzymes and others that catalyse halide oxidation using dioxygen, hydrogen peroxide and hydroperoxides, or that promote nucleophilic halide addition reactions. PMID- 19675647 TI - The diversity of type Ia supernovae from broken symmetries. AB - Type Ia supernovae result when carbon-oxygen white dwarfs in binary systems accrete mass from companion stars, reach a critical mass and explode. The near uniformity of their light curves makes these supernovae good 'standard candles' for measuring cosmic expansion, but a correction must be applied to account for the fact that the brighter ones have broader light curves. One-dimensional modelling, with a certain choice of parameters, can reproduce this general trend in the width-luminosity relation; but the processes of ignition and detonation have recently been shown to be intrinsically asymmetric, so parameterization must have its limits. Here we report multi-dimensional modelling of the explosion physics and radiative transfer, which reveals that the breaking of spherical symmetry is a critical factor in determining both the width-luminosity relation and the observed scatter about it. The deviation from spherical symmetry can also explain the finite polarization detected in the light from some supernovae. The slope and normalization of the width-luminosity relation has a weak dependence on certain properties of the white dwarf progenitor, in particular the trace abundances of elements other than carbon and oxygen. Failing to correct for this effect could lead to systematic overestimates of up to 2 per cent in the distance to remote supernovae. PMID- 19675646 TI - Design of functional metalloproteins. AB - Metalloproteins catalyse some of the most complex and important processes in nature, such as photosynthesis and water oxidation. An ultimate test of our knowledge of how metalloproteins work is to design new metalloproteins. Doing so not only can reveal hidden structural features that may be missing from studies of native metalloproteins and their variants, but also can result in new metalloenzymes for biotechnological and pharmaceutical applications. Although it is much more challenging to design metalloproteins than non-metalloproteins, much progress has been made in this area, particularly in functional design, owing to recent advances in areas such as computational and structural biology. PMID- 19675648 TI - Storms in the tropics of Titan. AB - Methane clouds, lakes and most fluvial features on Saturn's moon Titan have been observed in the moist high latitudes, while the tropics have been nearly devoid of convective clouds and have shown an abundance of wind-carved surface features like dunes. The presence of small-scale channels and dry riverbeds near the equator observed by the Huygens probe at latitudes thought incapable of supporting convection (and thus strong rain) has been suggested to be due to geological seepage or other mechanisms not related to precipitation. Here we report the presence of bright, transient, tropospheric clouds in tropical latitudes. We find that the initial pulse of cloud activity generated planetary waves that instigated cloud activity at other latitudes across Titan that had been cloud-free for at least several years. These observations show that convective pulses at one latitude can trigger short-term convection at other latitudes, even those not generally considered capable of supporting convection, and may also explain the presence of methane-carved rivers and channels near the Huygens landing site. PMID- 19675649 TI - Dense packings of the Platonic and Archimedean solids. AB - Dense particle packings have served as useful models of the structures of liquid, glassy and crystalline states of matter, granular media, heterogeneous materials and biological systems. Probing the symmetries and other mathematical properties of the densest packings is a problem of interest in discrete geometry and number theory. Previous work has focused mainly on spherical particles-very little is known about dense polyhedral packings. Here we formulate the generation of dense packings of polyhedra as an optimization problem, using an adaptive fundamental cell subject to periodic boundary conditions (we term this the 'adaptive shrinking cell' scheme). Using a variety of multi-particle initial configurations, we find the densest known packings of the four non-tiling Platonic solids (the tetrahedron, octahedron, dodecahedron and icosahedron) in three-dimensional Euclidean space. The densities are 0.782..., 0.947..., 0.904... and 0.836..., respectively. Unlike the densest tetrahedral packing, which must not be a Bravais lattice packing, the densest packings of the other non-tiling Platonic solids that we obtain are their previously known optimal (Bravais) lattice packings. Combining our simulation results with derived rigorous upper bounds and theoretical arguments leads us to the conjecture that the densest packings of the Platonic and Archimedean solids with central symmetry are given by their corresponding densest lattice packings. This is the analogue of Kepler's sphere conjecture for these solids. PMID- 19675650 TI - Atlantic hurricanes and climate over the past 1,500 years. AB - Atlantic tropical cyclone activity, as measured by annual storm counts, reached anomalous levels over the past decade. The short nature of the historical record and potential issues with its reliability in earlier decades, however, has prompted an ongoing debate regarding the reality and significance of the recent rise. Here we place recent activity in a longer-term context by comparing two independent estimates of tropical cyclone activity over the past 1,500 years. The first estimate is based on a composite of regional sedimentary evidence of landfalling hurricanes, while the second estimate uses a previously published statistical model of Atlantic tropical cyclone activity driven by proxy reconstructions of past climate changes. Both approaches yield consistent evidence of a peak in Atlantic tropical cyclone activity during medieval times (around ad 1000) followed by a subsequent lull in activity. The statistical model indicates that the medieval peak, which rivals or even exceeds (within uncertainties) recent levels of activity, results from the reinforcing effects of La-Nina-like climate conditions and relative tropical Atlantic warmth. PMID- 19675651 TI - Glacial effects limiting mountain height. AB - The height of mountain ranges reflects the balance between tectonic rock uplift, crustal strength and surface denudation. Tectonic deformation and surface denudation are interdependent, however, and feedback mechanisms-in particular, the potential link to climate-are subjects of intense debate. Spatial variations in fluvial denudation rate caused by precipitation gradients are known to provide first-order controls on mountain range width, crustal deformation rates and rock uplift. Moreover, limits to crustal strength are thought to constrain the maximum elevation of large continental plateaus, such as those in Tibet and the central Andes. There are indications that the general height of mountain ranges is also directly influenced by the extent of glaciation through an efficient denudation mechanism known as the glacial buzzsaw. Here we use a global analysis of topography and show that variations in maximum mountain height correlate closely with climate-controlled gradients in snowline altitude for many high mountain ranges across orogenic ages and tectonic styles. With the aid of a numerical model, we further demonstrate how a combination of erosional destruction of topography above the snowline by glacier-sliding and commensurate isostatic landscape uplift caused by erosional unloading can explain observations of maximum mountain height by driving elevations towards an altitude window just below the snowline. The model thereby self-consistently produces the hypsometric signature of the glacial buzzsaw, and suggests that differences in the height of mountain ranges mainly reflect variations in local climate rather than tectonic forces. PMID- 19675654 TI - [S.E.N. recommendations regarding transplanted patients returning to dialysis. Objectives and methods]. PMID- 19675652 TI - Homothallic and heterothallic mating in the opportunistic pathogen Candida albicans. AB - Candida albicans is the most common fungal pathogen in humans, causing both debilitating mucosal infections and potentially life-threatening systemic infections. Until recently, C. albicans was thought to be strictly asexual, existing only as an obligate diploid. A cryptic mating cycle has since been uncovered in which diploid a and alpha cells undergo efficient cell and nuclear fusion, resulting in tetraploid a/alpha mating products. Whereas mating between a and alpha cells has been established (heterothallism), we report here two pathways for same-sex mating (homothallism) in C. albicans. First, unisexual populations of a cells were found to undergo autocrine pheromone signalling and same-sex mating in the absence of the Bar1 protease. In both C. albicans and Saccharomyces cerevisiae, Bar1 is produced by a cells and inactivates mating pheromone alpha, typically secreted by alpha cells. C. albicans Deltabar1 a cells were shown to secrete both a and alpha mating pheromones; alpha-pheromone activated self-mating in these cells in a process dependent on Ste2, the receptor for alpha-pheromone. In addition, pheromone production by alpha cells was found to promote same-sex mating between wild-type a cells. These results establish that homothallic mating can occur in C. albicans, revealing the potential for genetic exchange even within unisexual populations of the organism. Furthermore, Bar1 protease has an unexpected but pivotal role in determining whether sexual reproduction can potentially be homothallic or is exclusively heterothallic. These findings also have implications for the mode of sexual reproduction in related species that propagate unisexually, and indicate a role for specialized sexual cycles in the survival and adaptation of pathogenic fungi. PMID- 19675655 TI - [Impact of advanced kidney disease on transplanted patients and their return to dialysis]. AB - Although long-term outcomes also improved, graft loss caused by chronic allograft nephropathy remains an important obstacle. This situation, together with the progressive increase in the number of renal transplant patients, means that the population of transplant patients readmitted to a dialysis program will be progressively greater. The mortality rate in patients starting dialysis after graft loss has been reported as variable, though higher than that observed in patients with a functioning graft and that observed in patients on dialysis treatment. However, it is not known how the management of chronic kidney disease patients in the transplant setting differs from that of patients with native kidney disease with a similar degree of renal dysfunction. Many patients in stages 4T-5T have chronic kidney disease related complications that fall below targets established for nontransplant chronic kidney disease patients. A limited number of studies have evaluated patients returning to dialysis after graft failure and the different guidelines in the setting of transplantation have not analyzed this crucial aspect so important. Parting from this premise, a working group of the Spanish Society of Nephrology in the field of kidney transplantation and dialysis has reviewed in-depth each of the clinical aspects of care of patients with kidney transplant failure coming back to dialysis and drawn up a consensus document in order to optimize the management of this condition. PMID- 19675656 TI - [Progression factors in chronic kidney disease. Immunological mechanisms]. AB - In kidney transplantation patient and graft survival are excellent in short-term and mid-term, although they remain stable in the long-term.The incidence of acute rejection has decreased to 8%-15%.Despite marked progress in understanding immunologic mechanisms involved in transplantation, new tools are required to detect early changes that could affect allograft function allowing us to anticipate histological lesions and providing a more accurate use of immunosuppressive drugs.From an immunologic point of view, efforts should be directed to avoid interstitial fibrosis and tubular atrophy (IF/TA) and to prevent antibody-mediated rejection.The most frequent cause of late graft loss is IF/TA.Improvement in kidney transplant results have been achieved with calcineurin inhibitors -CNI- (cyclosporin and tacrolimus), antiproliferative agents (mycophenolate mofetil and enteric-coated mycophenolic acid) and T-cell depleting antibodies. The combination of tacrolimus + mycophenolate mofetil + steroids has been the gold standard in kidney transplant immunosuppression. An adequate balance in order to maintain the appropriate immune response is essential to the patient to avoid infections or neoplasias as well to prevent rejection.In renal transplant recipients with chronic kidney disease stage 4T in which renal function remains stable,immuno-suppressive drugs can be continued at the usual maintenance doses. As GFR declines, CNI and antiproliferative drugs should be reduced. PMID- 19675657 TI - [Progression factors in chronic kidney disease. Non-immunological mechanisms]. AB - Non-immunological factors in the progression of kidney disease in transplant patients are the following: high blood pressure, proteinuria, dislypidemia, etc. 1. Arterial hypertension treatment: Blood pressure must be measured periodically in all transplant patients. Similarly to native kidneys, in renal transplant patients arterial hypertension is a risk factor in the progression of kidney disease. Arterial hypertension represent a clinical marker of chronic allograft nephropathy and contributes to graft loss and to the morbid- mortality of these patients (Evidence level C). Blood pressure control should be < 130/80 mm Hg for renal transplant patients without proteinuria and 125/75 mm Hg for proteinuric patients (> 1 g/24 hours). Hypertension and proteinuria are frequently associated in the same patients, a global treatment of both seems more rational (Evidence level C). General measures should be instigated first with pharmacological therapy. All antihypertensive drugs are useful in renal transplant patients and the majority of patients will need two or more drugs. In proteinuric patients an angiotensin receptor antagonist or an ACE-inhibitor should be initiated. It is advisable to monitor the serum potassium and creatinine after the start of this drugs or during the treatment periodically, especially in patients with chronic kidney disease stage IV-V. 2. Proteinuria treatment: Proteinuria has been strongly correlated with reduced function and graft survival. Lowering proteinuria to values as near to normal as possible (< 0.5 g/24 hours). To reduce proteinuria, an angiotensin receptor antagonist, an ACE-inhibitor or a combination of both are required, with serum potassium or creatinine monitoring, especially in patients with chronic kidney disease stage IV-V. 3. Dyslipidemia treatment: For kidney transplant recipients the assessment of dyslipidemias should include a complete fasting lipid profile with total cholesterol, LDL, HDL, and triglycerides. Evidence from the general population indicates that treatment of dyslipidemias reduces cardiovascular disease and evidence in kidney transplant patients suggests that judicious treatment can be safe and effective in improving dyslipidemia. Therapeutic goal must be LDL < 100 mg/dl. (Evidence level C). 4. Others: Cigarette smoking, glucose intolerance or diabetes control and obesity should be assessed. PMID- 19675658 TI - [Managing anaemia in kidney transplant patients with chronic kidney disease]. AB - DEFINITION: The definition of anemia is established by the World Health Organization and was subsequently adopted by the American Society of Transplantation, which defines anemia as hemoglobin concentration <12 g/dl in women and <13 g/dl in men. PREVALENCE OF ANEMIA POSTTRANSPLANTATION: Varies throughout the posttransplantation period and is associated with the degree of renal graft function. The relationship between hemoglobin levels and glomerular filtration does not behave the same way as in the population with chronic kidney disease. The results of various studies show a high prevalence in the first months after transplantation (<6 months), which decreases from the first year posttransplantation and then increases related to loss of graft function. European study on the management of anemia showed a prevalence of anemia in 38.6% and only 18% of patients with severe anemia were treated with erythropoietin (EPO). PATHOPHYSIOLOGY: There is a decrease in the synthesis of erythropoietin (EPO) or an increase in resistance to EPO. There are many factors that can cause anemia post-transplantation. Some of these factors are specific to transplanted patients whilst others are common to all patients with chronic kidney disease. Among the common factors there are: the degree of renal function and iron deficiency and among the factors of transplantation there are acute rejection, post-transplantation medications, infections and malignancies. CLINICAL RESULTS: The available data evaluating the association of anemia with morbidity and mortality of the patient and graft survival are scarce. Most studies are retrospective and analyze experiences of individual centers. They showed a higher mortality and morbidity among patients with a hemoglobin <11 g/dl. (Evidence B). TREATMENT OF POST-TRANSPLANTATION ANEMIA: Erythropoiesis-stimulating agents (ESA) and replenishment of iron deposits (Evidence A). RESPONSE TO TREATMENT: In transplant patients there may be some resistance to treatment with erythropoiesis stimulating agents (ESA) due to the use of myelosuppressive medications, chronic inflammation and other factors. - Adverse effects of treatment with ESA: There are few controlled studies failed to show respect to the ESA that are effective and unlikely to accelerate the deterioration of renal function but may aggravate hypertension. PMID- 19675659 TI - [Changes in bone and mineral metabolism in kidney transplant patients with chronic kidney disease]. AB - DESCRIPTION: Recently, the Foundation has proposed new definitions KDIGO to refer to the alterations of bone - mineral metabolism in patients with chronic renal disease (CRD), relegating the traditional term of renal osteodystrophy (ODR). RECOMMEND: The term ODR exclusively to define alterations in bone morphology and architecture characteristic of the ERC. - And the term of bone-mineral alteration associated with the CRD to describe biochemical changes, and skeletal calcifications that occur as a result of alterations in mineral metabolism in the CRD. PATHOPHYSIOLOGY: The different metabolic abnormalities are secondary to the progressive loss of renal mass and renal function that leads to retention of phosphorus and a decrease in the levels of calcitriol which are responsible for the skeletal resistance to the action of PTH. CLINICAL FEATURES: The main clinical manifestations of abnormal bone mineral metabolism are posttransplantation osteoporosis and osteopenia producing an increase in fractures, osteonecrosis, and bone pain. DIAGNOSTIC METHODS: Biochemical parameters (calcium, phosphorus, PTH, 25 hydroxyvitamin D), X-ray bone densitometry and bone biopsy. (Evidence B). THERAPEUTIC ALTERNATIVES: It is recommended for the treatment and prevention of osteopenia - osteoporosis in transplant patients based on data from clinical evidence available from other study populations, such as in patients with chronic kidney disease. In addition to specific treatment, we must take into account the preventive measures to reduce the risk of fractures. Treatment includes specific measures for the prevention of bone loss (active metabolite of vitamin D analogues and bisphosphonates) and the treatment of persistent hyperparathyroidism (calcimimeticos). (Evidence B). PMID- 19675660 TI - [Evaluating renal function and indications for starting dialysis]. AB - EVALUATION OF THE RENAL FUNCTION: For the follow-up of the graft renal function it must be measured the glomerular filtration rate by means of formulae that use the serum creatinine. The most used equation is the brief formula MDRD. - All patients transplanted must be included in the group of Renal Chronic Disease though the glomerular filtration rate is normal and there is no evidence of renal damage. - The measures of intervention proposed in the classification of the Renal Chronic Disease for its progressive establishment in the stage 1 to 3, must be applied to all the transplanted THE BEGINNING OF DIALYSIS: In spite of receiving attention of Nephrologists along the whole evolution, the patients with chronic dysfunction of the graft that need treatment with dialysis start later and with more uremic complications that the patients who start dialysis for the first time. - To change this trend, it is necessary to consider the treatment with dialysis when the glomerular filtration rate is lower than 15 ml/min/1,73 m2. If there appears any complication related to the uremia that cannot be handled by conservative treatment, the beginning of the dialysis is necessary. THE BEGINNING OF THE DIALYSIS OF PROGRAMMED FORM: The beginning of the dialysis of not programmed form in transplanted patients is difficult to justify if we take into account that such patients, have received nephrological attention along all their evolution. - To get a new vascular access in these patients can be difficult depending on the previous trombosis of arteriovenous fistulas. Therefore, it must be realized a prompt evaluation for de department of vascular surgery to guarantee a suitable vascular access. - As general norm, one must follow the same criterion advised for the not transplanted patient: the vascular access must be considered when the glomerular filtration rate is lower than 20 m/min/1,73 m2. - The patient who is going to be treated by dialysis peritoneal precise a very narrow follow-up to be able to programme the placement of the catheter peritoneal with a minimum of 15 days before beginning the training. PMID- 19675661 TI - [Choosing the dialysis method for kidney transplant patients with advanced kidney disease]. AB - The choice of the most of dialysis modality after renal graft loss is an unanswered question. Most patients start hemodialysis (HD) in this situation, because of several reasons: 1. In most dialysis programs HD predominates clearly over Peritoneal Dialysis (PD). 2. The star of dialysis in emergency situations makes the physician use HD 3. The fear of infections in case of maintenance of immunosupression to avoid immune response and to keep residual renal function in case of PD. A lot of patients could undergo PD in order to maintain the previous style of life, as an alternative in case of absence of vascular access and to avoid vascular accesses in children. Mortality seems to be greater in patients with graft loss than in those who start dialysis for the first time, although the comparison between both groups is methodologically difficult. However, there is no difference in mortality between patients who start HD and those who start PD. Studies comparing the rate of peritonitis in PD patients in both groups find controversial results. The analysis of the few, retrospective and biased studies which look for differences in patient survival in HD and DP suggests that prognosis of both groups is similar. The choice of dialysis modality must be similar to those patients which begin dialysis for the first time. PMID- 19675662 TI - [Definitive removal of immunosuppressors]. AB - There's no controlled and prospective studies which show the safest and most effective way to reduce or suspend immunosuppression drugs dosage, and only few groups have published their own protocols. There are reasons to discontinue the immunosuppression therapy; the high incidence of infections is the most important. However, a fast withdrawal is not free of problems, like are residual renal function decline and graft intolerance signs, which could take to nephrectomy, a high risk intervention. Most recommended guidelines for immunosuppression use are: - Antimetabolites immediately cancellation and corticoesteroids slow drop (level C recommendation). - Calcineurin inhibitors could be discontinued but if residual renal function is still significant, it is recommended to maintain a low dosage over three to six months; then, withdrawal may be done slowly (level C recommendation). We have not found information supporting the immunosuppression use beyond six months. PMID- 19675663 TI - [Managing a failed kidney graft. Nephrectomy versus embolisation]. AB - The early diagnosis of the graft intolerance syndrome or a subclinical state of chronic inflammation due to a failed kidney allograft, is one of the goals that the nephrologists must fulfill to take a series of measures directed to solve this situation. Fever, haematuria, local pain and/or tenderness are the main clinical criteria to make a diagnosis. However, oftenly there are not any clinical symptoms and only the presence of parameters of chronic inflammation (elevated C-reactive protein, erythrocyte sedimentation rate, hypoalbuminemia and anemia resistant to erythropoietin therapy) are signs of this entity. Maintenance of immunosuppressive treatment is not advisable due to the risk of infections as well as the increase in cardiovascular risk (level evidence A). Transplantectomy is the best treatment if there are some associated complications such as allograft infection, neoplasia or high risk of graft rupture. However, surgical treatment is not exempt from risks and it is associated to a considerable rate of complications, with the consequent prolongation of the hospitalization stay. Therefore it is desirable to use less invasive procedures, such as embolization. This could be the first step unless the conditions enumerated in point 3 come up (Level evidence B). It is desirable to use prophylactic antibiotic before the embolization to avoid infectious complications (Level evidence B). PMID- 19675664 TI - [Retransplant]. AB - Despite the advances in immunosuppressive therapy and in patient care, about 20 30% of patients will have lost their grafts after 3 years and this loss will continue by 3-4% per year. These patients are included in maintenance dialysis programmes and account for 4 to 10% of those admitted every year for maintenance dialysis therapy. Among those patients who loss their grafts 40-60% are included in transplant waiting lists. This increases the number of patients waiting for a graft and raises the dilemma about the rights to be included in deceased donor programmes. A common characteristic of these patients waiting for a second or even third transplant is the presence in the blood of antibodies to HLA antigens. A new transplant is the best therapeutic option for these patients, and the results are quite close to those achieved for the first graft. Moreover, a new transplant improves patient outcome when compared with those remaining in the waiting list. The best results are obtained in diabetic patients and in those between 18 to 50 years old (Evidence C). However, the percentage of patients retransplanted has not varied in the last years, possibly due to the wider criteria adopted on candidate selection that increases the waiting lists (Evidence B). In the second transplant, mismatched HLA-A,B antigens could be repeated if the recipient has not developed specific antibodies to these antigens. Recent cross-match has to be negative. Immunosuppressive therapy is similar to that used with first transplants. Lymphoproliferative diseases, BK virus nephropathy and primary glomerulonephritis do not preclude a second transplant (Evidence C). PMID- 19675665 TI - [Coordinating between the renal transplant unit and the non-transplant nephrology department]. AB - Patients with stabilized kidney transplant receive optimal management care when there is effective coordination between the transplant centre and the community nephrologist (Evidence level C). A good coordination with regular interactive communication between the transplant centre and community nephrologist is very positive for patients and beneficial to the transplant centre and community nephrologist (Evidence level C). Many of the clinical objectives for management of kidney transplant recipients are similar to those related to chronic kidney disease patients (Evidence level C). A good coordination between the transplant centre and community nephrologist needs organizational requirements and clinical management protocols (Evidence level C).When irreversible renal allograft failure occurs, the community nephrologist must assume the preparation for dialysis as with other patients with advanced chronic kidney disease: choose dialysis methods, create arteriovenous fistulae or place peritoneal catheter and identify dialysis treatment centre. Moreover, the transplant centre and the community nephrologist will jointly decide the best moment to start dialysis or the possibility of preemptive kidney transplant (Evidence level C). PMID- 19675667 TI - Missing data in randomized clinical trials for weight loss: scope of the problem, state of the field, and performance of statistical methods. AB - BACKGROUND: Dropouts and missing data are nearly-ubiquitous in obesity randomized controlled trails, threatening validity and generalizability of conclusions. Herein, we meta-analytically evaluate the extent of missing data, the frequency with which various analytic methods are employed to accommodate dropouts, and the performance of multiple statistical methods. METHODOLOGY/PRINCIPAL FINDINGS: We searched PubMed and Cochrane databases (2000-2006) for articles published in English and manually searched bibliographic references. Articles of pharmaceutical randomized controlled trials with weight loss or weight gain prevention as major endpoints were included. Two authors independently reviewed each publication for inclusion. 121 articles met the inclusion criteria. Two authors independently extracted treatment, sample size, drop-out rates, study duration, and statistical method used to handle missing data from all articles and resolved disagreements by consensus. In the meta-analysis, drop-out rates were substantial with the survival (non-dropout) rates being approximated by an exponential decay curve (e(-lambdat)) where lambda was estimated to be .0088 (95% bootstrap confidence interval: .0076 to .0100) and t represents time in weeks. The estimated drop-out rate at 1 year was 37%. Most studies used last observation carried forward as the primary analytic method to handle missing data. We also obtained 12 raw obesity randomized controlled trial datasets for empirical analyses. Analyses of raw randomized controlled trial data suggested that both mixed models and multiple imputation performed well, but that multiple imputation may be more robust when missing data are extensive. CONCLUSION/SIGNIFICANCE: Our analysis offers an equation for predictions of dropout rates useful for future study planning. Our raw data analyses suggests that multiple imputation is better than other methods for handling missing data in obesity randomized controlled trials, followed closely by mixed models. We suggest these methods supplant last observation carried forward as the primary method of analysis. PMID- 19675666 TI - How thioredoxin dissociates its mixed disulfide. AB - The dissociation mechanism of the thioredoxin (Trx) mixed disulfide complexes is unknown and has been debated for more than twenty years. Specifically, opposing arguments for the activation of the nucleophilic cysteine as a thiolate during the dissociation of the complex have been put forward. As a key model, the complex between Trx and its endogenous substrate, arsenate reductase (ArsC), was used. In this structure, a Cys29(Trx)-Cys89(ArsC) intermediate disulfide is formed by the nucleophilic attack of Cys29(Trx) on the exposed Cys82(ArsC) Cys89(ArsC) in oxidized ArsC. With theoretical reactivity analysis, molecular dynamics simulations, and biochemical complex formation experiments with Cys mutants, Trx mixed disulfide dissociation was studied. We observed that the conformational changes around the intermediate disulfide bring Cys32(Trx) in contact with Cys29(Trx). Cys32(Trx) is activated for its nucleophilic attack by hydrogen bonds, and Cys32(Trx) is found to be more reactive than Cys82(ArsC). Additionally, Cys32(Trx) directs its nucleophilic attack on the more susceptible Cys29(Trx) and not on Cys89(ArsC). This multidisciplinary approach provides fresh insights into a universal thiol/disulfide exchange reaction mechanism that results in reduced substrate and oxidized Trx. PMID- 19675668 TI - No evidence for mutations of CTCFL/BORIS in Silver-Russell syndrome patients with IGF2/H19 imprinting control region 1 hypomethylation. AB - BACKGROUND: Silver-Russell syndrome (SRS) is a genetically and clinically heterogeneous disease. Although no protein coding gene defects have been reported in SRS patients, approximately 50% of SRS patients carry epimutations (hypomethylation) at the IGF2/H19 imprinting control region 1 (ICR1). Proper methylation at ICR1 is crucial for the imprinted expression of IGF2, a fetal growth factor. CTCFL, a testis-specific protein, has recently been proposed to play a role in the establishment of DNA methylation at the murine equivalent of ICR1. A screen was undertaken to assess whether CTCFL is mutated in SRS patients with hypomethylation, to explore a link between the observed epimutations and a genetic cause of the disease. METHODOLOGY/PRINCIPAL FINDINGS: DNA was obtained from 36 SRS patients with hypomethylation at ICR1. All CTCFL coding exons were sequenced and analyzed for duplications/deletions using both multiplex ligation dependent probe amplification, with a custom CTCFL probe set, and genomic qPCR. Novel SNP alleles were analyzed for potential differential splicing in vitro utilizing a splicing assay. Neither mutations of CTCFL nor duplications/deletions were observed. Five novel SNPs were identified and have been submitted to dbSNP. In silico splice prediction suggested one novel SNP, IVS2-66A>C, activated a cryptic splice site, resulting in aberrant splicing and premature termination. In vitro splicing assays did not confirm predicted aberrant splicing. CONCLUSIONS/SIGNIFICANCE: As no mutations were detected at CTCFL in the patients examined, we conclude that genetic alterations of CTCFL are not responsible for the SRS hypomethylation. We suggest that analysis of other genes involved in the establishment of DNA methylation at imprinted genes, such as DNMT3A and DNMT3L, may provide insight into the genetic cause of hypomethylation in SRS patients. PMID- 19675669 TI - The diagnostic and prognostic accuracy of five markers of serious bacterial infection in Malawian children with signs of severe infection. AB - BACKGROUND: Early recognition and prompt and appropriate antibiotic treatment can significantly reduce mortality from serious bacterial infections (SBI). The aim of this study was to evaluate the utility of five markers of infection: C reactive protein (CRP), procalcitonin (PCT), soluble triggering receptor expressed on myeloid cells-1 (sTREM-1), CD163 and high mobility group box-1 (HMGB1), as markers of SBI in severely ill Malawian children. METHODOLOGY AND PRINCIPAL FINDINGS: Children presenting with a signs of meningitis (n = 282) or pneumonia (n = 95), were prospectively recruited. Plasma samples were taken on admission for CRP, PCT, sTREM-1 CD163 and HMGB1 and the performance characteristics of each test to diagnose SBI and to predict mortality were determined. Of 377 children, 279 (74%) had SBI and 83 (22%) died. Plasma CRP, PCT, CD163 and HMGB1 and were higher in HIV-infected children than in HIV uninfected children (p<0.01). In HIV-infected children, CRP and PCT were higher in children with SBI compared to those with no detectable bacterial infection (p<0.0005), and PCT and CD163 were higher in non-survivors (p = 0.001, p = 0.05 respectively). In HIV-uninfected children, CRP and PCT were also higher in children with SBI compared to those with no detectable bacterial infection (p<0.0005), and CD163 was higher in non-survivors (p = 0.05). The best predictors of SBI were CRP and PCT, and areas under the curve (AUCs) were 0.81 (95% CI 0.73 0.89) and 0.86 (95% CI 0.79-0.92) respectively. The best marker for predicting death was PCT, AUC 0.61 (95% CI 0.50-0.71). CONCLUSIONS: Admission PCT and CRP are useful markers of invasive bacterial infection in severely ill African children. The study of these markers using rapid tests in a less selected cohort would be important in this setting. PMID- 19675670 TI - Emergence of community-associated methicillin-resistant Staphylococcus aureus associated with pediatric infection in Cambodia. AB - BACKGROUND: The incidence of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) infection is rising in the developed world but appears to be rare in developing countries. One explanation for this difference is that resource poor countries lack the diagnostic microbiology facilities necessary to detect the presence of CA-MRSA carriage and infection. METHODOLOGY AND PRINCIPAL FINDINGS: We developed diagnostic microbiology capabilities at the Angkor Hospital for Children, Siem Reap, western Cambodia in January 2006 and in the same month identified a child with severe community-acquired impetigo caused by CA-MRSA. A study was undertaken to identify and describe additional cases presenting between January 2006 and December 2007. Bacterial isolates underwent molecular characterization using multilocus sequence typing, staphylococcal cassette chromosome mec (SCCmec) typing, and PCR for the presence of the genes encoding Panton-Valentine Leukocidin (PVL). Seventeen children were identified with CA-MRSA infection, of which 11 had skin and soft tissue infection and 6 had invasive disease. The majority of cases were unrelated in time or place. Molecular characterization identified two independent MRSA clones; fifteen isolates were sequence type (ST) 834, SCCmec type IV, PVL gene-negative, and two isolates were ST 121, SCCmec type V, PVL gene-positive. CONCLUSIONS: This represents the first ever report of MRSA in Cambodia, spread of which would pose a significant threat to public health. The finding that cases were mostly unrelated in time or place suggests that these were sporadic infections in persons who were CA-MRSA carriers or contacts of carriers, rather than arising in the context of an outbreak. PMID- 19675671 TI - Vitamin D status is positively correlated with regulatory T cell function in patients with multiple sclerosis. AB - BACKGROUND: In several autoimmune diseases, including multiple sclerosis (MS), a compromised regulatory T cell (Treg) function is believed to be critically involved in the disease process. In vitro, the biologically active metabolite of vitamin D has been shown to promote Treg development. A poor vitamin D status has been linked with MS incidence and MS disease activity. In the present study, we assess a potential in vivo correlation between vitamin D status and Treg function in relapsing remitting MS (RRMS) patients. METHODOLOGY/PRINCIPAL FINDINGS: Serum levels of 25-hydroxyvitamin D (25(OH)D) were measured in 29 RRMS patients. The number of circulating Tregs was assessed by flow-cytometry, and their functionality was tested in vitro in a CFSE-based proliferation suppression assay. Additionally, the intracellular cytokine profile of T helper cells was determined directly ex-vivo by flow-cytometry. Serum levels of 25(OH)D correlated positively with the ability of Tregs to suppress T cell proliferation (R = 0.590, P = 0.002). No correlation between 25(OH)D levels and the number of Tregs was found. The IFN-gamma/IL-4 ratio (Th1/Th2-balance) was more directed towards IL-4 in patients with favourable 25(OH)D levels (R = -0.435, P = 0.023). CONCLUSIONS/SIGNIFICANCE: These results show an association of high 25(OH)D levels with an improved Treg function, and with skewing of the Th1/Th2 balance towards Th2. These findings suggest that vitamin D is an important promoter of T cell regulation in vivo in MS patients. It is tempting to speculate that our results may not only hold for MS, but also for other autoimmune diseases. Future intervention studies will show whether modulation of vitamin D status results in modulation of the T cell response and subsequent amelioration of disease activity. PMID- 19675672 TI - Selective interaction of syntaxin 1A with KCNQ2: possible implications for specific modulation of presynaptic activity. AB - KCNQ2/KCNQ3 channels are the molecular correlates of the neuronal M-channels, which play a major role in the control of neuronal excitability. Notably, they differ from homomeric KCNQ2 channels in their distribution pattern within neurons, with unique expression of KCNQ2 in axons and nerve terminals. Here, combined reciprocal coimmunoprecipitation and two-electrode voltage clamp analyses in Xenopus oocytes revealed a strong association of syntaxin 1A, a major component of the exocytotic SNARE complex, with KCNQ2 homomeric channels resulting in a approximately 2-fold reduction in macroscopic conductance and approximately 2-fold slower activation kinetics. Remarkably, the interaction of KCNQ2/Q3 heteromeric channels with syntaxin 1A was significantly weaker and KCNQ3 homomeric channels were practically resistant to syntaxin 1A. Analysis of different KCNQ2 and KCNQ3 chimeras and deletion mutants combined with in-vitro binding analysis pinpointed a crucial C-terminal syntaxin 1A-association domain in KCNQ2. Pull-down and coimmunoprecipitation analyses in hippocampal and cortical synaptosomes demonstrated a physical interaction of brain KCNQ2 with syntaxin 1A, and confocal immunofluorescence microscopy showed high colocalization of KCNQ2 and syntaxin 1A at presynaptic varicosities. The selective interaction of syntaxin 1A with KCNQ2, combined with a numerical simulation of syntaxin 1A's impact in a firing-neuron model, suggest that syntaxin 1A's interaction is targeted at regulating KCNQ2 channels to fine-tune presynaptic transmitter release, without interfering with the function of KCNQ2/3 channels in neuronal firing frequency adaptation. PMID- 19675673 TI - Identification of a putative Crf splice variant and generation of recombinant antibodies for the specific detection of Aspergillus fumigatus. AB - BACKGROUND: Aspergillus fumigatus is a common airborne fungal pathogen for humans. It frequently causes an invasive aspergillosis (IA) in immunocompromised patients with poor prognosis. Potent antifungal drugs are very expensive and cause serious adverse effects. Their correct application requires an early and specific diagnosis of IA, which is still not properly achievable. This work aims to a specific detection of A. fumigatus by immunofluorescence and the generation of recombinant antibodies for the detection of A. fumigatus by ELISA. RESULTS: The A. fumigatus antigen Crf2 was isolated from a human patient with proven IA. It is a novel variant of a group of surface proteins (Crf1, Asp f9, Asp f16) which belong to the glycosylhydrolase family. Single chain fragment variables (scFvs) were obtained by phage display from a human naive antibody gene library and an immune antibody gene library generated from a macaque immunized with recombinant Crf2. Two different selection strategies were performed and shown to influence the selection of scFvs recognizing the Crf2 antigen in its native conformation. Using these antibodies, Crf2 was localized in growing hyphae of A. fumigatus but not in spores. In addition, the antibodies allowed differentiation between A. fumigatus and related Aspergillus species or Candida albicans by immunofluorescence microscopy. The scFv antibody clones were further characterized for their affinity, the nature of their epitope, their serum stability and their detection limit of Crf2 in human serum. CONCLUSION: Crf2 and the corresponding recombinant antibodies offer a novel approach for the early diagnostics of IA caused by A. fumigatus. PMID- 19675674 TI - GSK3beta is involved in JNK2-mediated beta-catenin inhibition. AB - BACKGROUND: We have recently reported that mitogen-activated protein kinase (MAPK) JNK1 downregulates beta-catenin signaling and plays a critical role in regulating intestinal homeostasis and in suppressing tumor formation. This study was designed to determine whether JNK2, another MAPK, has similar and/or different functions in the regulation of beta-catenin signaling. METHODOLOGY AND PRINCIPAL FINDINGS: We used an in vitro system with manipulation of JNK2 and beta catenin expression and found that activated JNK2 increased GSK3beta activity and inhibited beta-catenin expression and transcriptional activity. However, JNK2 mediated downregulation of beta-catenin was blocked by the proteasome inhibitor MG132 and GSK3beta inhibitor lithium chloride. Moreover, targeted mutations at GSK3beta phosphorylation sites (Ser33 and Ser37) of beta-catenin abrogated JNK2 mediated suppression of beta-catenin. In vivo studies further revealed that JNK2 deficiency led to upregulation of beta-catenin and increase of GSK3-beta phosphorylation in JNK2-/- mouse intestinal epithelial cells. Additionally, physical interaction and co-localization among JNK2, beta-catenin and GSK3beta were observed by immunoprecipitation, mammalian two-hybridization assay and confocal microscopy, respectively. CONCLUSION AND SIGNIFICANCE: In general, our data suggested that JNK2, like JNK1, interacts with and suppresses beta-catenin signaling in vitro and in vivo, in which GSK3beta plays a key role, although previous studies have shown distinct functions of JNK1 and JNK2. Our study also provides a novel insight into the crosstalk between Wnt/beta-catenin and MAPK JNKs signaling. PMID- 19675675 TI - The impact of IPTi and IPTc interventions on malaria clinical burden - in silico perspectives. AB - BACKGROUND: Clinical management of malaria is a major health issue in sub-Saharan Africa. New strategies based on intermittent preventive treatment (IPT) can tackle disease burden by simultaneously reducing frequency of infections and life threatening illness in infants (IPTi) and children (IPTc), while allowing for immunity to build up. However, concerns as to whether immunity develops efficiently in treated individuals, and whether there is a rebound effect after treatment is halted, have made it imperative to define the effects that IPTi and IPTc exert on the clinical malaria scenario. METHODS AND FINDINGS: Here, we simulate several schemes of intervention under different transmission settings, while varying immunity build up assumptions. Our model predicts that infection risk and effectiveness of acquisition of clinical immunity under prophylactic effect are associated to intervention impact during treatment and follow-up periods. These effects vary across regions of different endemicity and are highly correlated with the interplay between the timing of interventions in age and the age dependent risk of acquiring an infection. However, even when significant rebound effects are predicted to occur, the overall intervention impact is positive. CONCLUSIONS: IPTi is predicted to have minimal impact on the acquisition of clinical immunity, since it does not interfere with the occurrence of mild infections, thus failing to reduce the underlying force of infection. On the contrary, IPTc has a significant potential to reduce transmission, specifically in areas where it is already low to moderate. PMID- 19675676 TI - A comparative neuroanatomical study of the red nucleus of the cat, macaque and human. AB - BACKGROUND: The human red nucleus (Nr) is comparatively less well-studied than that of cats or monkeys. Given the functional importance of reticular and midbrain structures in control of movement and locomotion as well as from an evolutionary perspective, we investigated the nature and extent of any differences in Nr projections to the olivary complex in quadrupedal and bipedal species. Using neuroanatomical tract-tracing techniques we developed a "neural sheet" hypothesis allowing us to propose how rubro-olivary relations differ among the three species. METHODS AND FINDINGS: Wheat germ agglutinin-horseradish peroxidase staining supports findings that the cat's nucleus accessories medialis of Bechtrew (NB) projects mainly to the lateral bend of the principal olive. We clarified boundaries among nucleus of Darkschewitsch (ND), NB and parvicellular red nucleus (pNr) of the cat's neural sheet. The macaque's ND-medial accessory olivary projection is rostro-caudally organized and the dorsomedial and ventrolateral parts of the macaque's pNr may project to the principal olive's rostral and caudal dorsal lamella; in cat it projects as well to pNr. Myelin- and Nissl-stained sections show that a well-developed dorsomedial part of the human Nr consists of densely packed cells, deriving small myelinated fibers that continue into the medial central tegmental tract. CONCLUSIONS: Based on these findings we suggest there are distinct bipedal-quadrupedal differences for Nr projections to the olivary complex. We propose the Nr of cats and monkeys comprise the ND, NB and pNr in a zonal sheet-like structure, retaining clear nuclear boundaries and an isolated, well-developed mNr. The human NB may be distinguished from its more specialised ND (ND lies alongside a well-developed pNr) in the human central gray. Phylogenetically, the NB may have been translocated into a roll-shaped Nr in the reticular formation, the dorsomedial portion of which might correspond to the cat's and monkey's NB. PMID- 19675677 TI - A nanoconjugate Apaf-1 inhibitor protects mesothelial cells from cytokine-induced injury. AB - BACKGROUND: Inflammation may lead to tissue injury. We have studied the modulation of inflammatory milieu-induced tissue injury, as exemplified by the mesothelium. Peritoneal dialysis is complicated by peritonitis episodes that cause loss of mesothelium. Proinflammatory cytokines are increased in the peritoneal cavity during peritonitis episodes. However there is scarce information on the modulation of cell death by combinations of cytokines and on the therapeutic targets to prevent desmesothelization. METHODOLOGY: Human mesothelial cells were cultured from effluents of stable peritoneal dialysis patients and from omentum of non-dialysis patients. Mesothelial cell death was studied in mice with S. aureus peritonitis and in mice injected with tumor necrosis factor alpha and interferon gamma. Tumor necrosis factor alpha and interferon gamma alone do not induce apoptosis in cultured mesothelial cells. By contrast, the cytokine combination increased the rate of apoptosis 2 to 3-fold over control. Cell death was associated with the activation of caspases and a pancaspase inhibitor prevented apoptosis. Specific caspase-8 and caspase-3 inhibitors were similarly effective. Co-incubation with both cytokines also impaired mesothelial wound healing in an in vitro model. However, inhibition of caspases did not improve wound healing and even impaired the long-term recovery from injury. By contrast, a polymeric nanoconjugate Apaf-1 inhibitor protected from apoptosis and allowed wound healing and long-term recovery. The Apaf-1 inhibitor also protected mesothelial cells from inflammation-induced injury in vivo in mice. CONCLUSION: Cooperation between tumor necrosis factor alpha and interferon gamma contributes to mesothelial injury and impairs the regenerative capacity of the monolayer. Caspase inhibition attenuates mesothelial cell apoptosis but does not facilitate regeneration. A drug targeting Apaf-1 allows protection from apoptosis as well as regeneration in the course of inflammation induced tissue injury. PMID- 19675678 TI - Molecular plasticity of E-cadherin and sialyl lewis x expression, in two comparative models of mammary tumorigenesis. AB - BACKGROUND: The process of metastasis involves a series of steps and interactions between the tumor embolus and the microenvironment. Key alterations in adhesion molecules are known to dictate progression from the invasive to malignant phenotype followed by colonization at a distant site. The invasive phenotype results from the loss of expression of the E-cadherin adhesion molecule, whereas the malignant phenotype is associated with an increased expression of the carbohydrate ligand-binding epitopes, (e.g. Sialyl Lewis (x/a)) that bind endothelial E-selectin of the lymphatics and vasculature. METHODOLOGY: Our study analyzed the expression of two adhesion molecules, E-cadherin and Sialyl Lewis x (sLe(x)), in both a canine mammary carcinoma and human inflammatory breast cancer (IBC) model, using double labelled immunofluorescence staining. RESULTS: Our results demonstrate that canine mammary carcinoma and human IBC exhibit an inversely correlated cellular expression of E-cadherin and sLe(x) within the same tumor embolus. CONCLUSIONS: Our results in these two comparative models (canine and human) suggest the existence of a biologically coordinated mechanism of E cadherin and sLe(x) expression (i.e. molecular plasticity) essential for tumor establishment and metastatic progression. PMID- 19675679 TI - Nonlinear time series analysis of nodulation factor induced calcium oscillations: evidence for deterministic chaos? AB - Legume plants form beneficial symbiotic interactions with nitrogen fixing bacteria (called rhizobia), with the rhizobia being accommodated in unique structures on the roots of the host plant. The legume/rhizobial symbiosis is responsible for a significant proportion of the global biologically available nitrogen. The initiation of this symbiosis is governed by a characteristic calcium oscillation within the plant root hair cells and this signal is activated by the rhizobia. Recent analyses on calcium time series data have suggested that stochastic effects have a large role to play in defining the nature of the oscillations. The use of multiple nonlinear time series techniques, however, suggests an alternative interpretation, namely deterministic chaos. We provide an extensive, nonlinear time series analysis on the nature of this calcium oscillation response. We build up evidence through a series of techniques that test for determinism, quantify linear and nonlinear components, and measure the local divergence of the system. Chaos is common in nature and it seems plausible that properties of chaotic dynamics might be exploited by biological systems to control processes within the cell. Systems possessing chaotic control mechanisms are more robust in the sense that the enhanced flexibility allows more rapid response to environmental changes with less energetic costs. The desired behaviour could be most efficiently targeted in this manner, supporting some intriguing speculations about nonlinear mechanisms in biological signaling. PMID- 19675680 TI - Effects of Beta-Blocker Titration on Glucose Homeostasis in Heart Failure. AB - BACKGROUND: Abnormal glucose metabolism and insulin resistance have been associated with heart failure incidence, severity, and mortality. Metabolic parameters such as hepatic glucose production may be altered by beta-adrenoceptor antagonists in patients with heart failure. OBJECTIVE: This study evaluated the effects of metoprolol or carvedilol up-titration on fasting glucose, insulin resistance and beta(2)-mediated glucose production in patients with chronic heart failure. METHODS: This was a prospective, randomized, active comparator study in 15 patients with AHA/ACC Stage C systolic dysfunction HF stable on medical therapy. Participants were randomized to metoprolol 25mg daily or carvedilol 3.125mg twice daily. Metoprolol was titrated to a target of 200mg daily, and carvedilol was titrated to 25mg twice daily over 8weeks. Insulin resistance as assessed by the homeostatic model, and terbutaline-induced glucose production (AUC(0-180)), were assessed at baseline and at 4 subsequent beta-blocker titration visits over 8 weeks. RESULTS: In all 15 patients, terbutaline-induced glucose AUC(0-180) decreased (p=0.0006) as beta-blocker doses increased. A significant reduction in glucose AUC(0-180) compared to baseline was only noted in patients taking metoprolol at 100mg daily (-2424.6 [95% CI -372.6 to -4478.4] mg/dL*min) and 200mg daily (-2437.2 [95% CI -15.1 to -4604.4] mg/dL*min), and not observed in those taking carvedilol. After beta-blocker titration, fasting glucose concentrations for the metoprolol and carvedilol groups were 86.9 (95% CI 89.8-101.6) mg/dL and 95.7 (95% CI 89.8-101.6) mg/dL, respectively (p=0.0273), adjusted for baseline values. There was no significant difference between metoprolol and carvedilol on insulin resistance. CONCLUSION: Increasing doses of beta-blockers are associated with decreased in beta2-mediated glucose production in heart failure. Metoprolol, but not carvedilol, decreases hepatic glucose production at commonly used heart failure doses. PMID- 19675682 TI - On Identifying Minimum Efficacious Doses in Combination Drug Trials. AB - To determine that a combination drug is effective, the FDA requires demonstration that the combination itself is effective and that each component makes a contribution to the claimed effect. In a study with at least one component known to be effective at the considered doses, these regulatory requirements can be satisfied by showing that the combination is superior to each component. We term such a combination an efficacious combination. In a dose-response study involving combination drugs, one of which is known to be effective, we are interested in detecting those combinations which are efficacious, and for which no lower dose combination is also efficacious. We term these combinations the minimum efficacious dose set and our goal is to estimate this set. Our procedure requires first identifying all possible minimum efficacious dose sets and the corresponding hypotheses for a given design. Next, the proper testing order based on a graph representation is established and the hypotheses are tested using the "average" test under the closed testing principle. This procedure is shown to have strong control of overall error rate. The power of this procedure is studied by simulation. PMID- 19675681 TI - Value of clinician assessment of hemodynamics in advanced heart failure: the ESCAPE trial. AB - BACKGROUND: We determined whether estimated hemodynamics from history and physical examination (H&P) reflect invasive measurements and predict outcomes in advanced heart failure (HF). The role of the H&P in medical decision making has declined in favor of diagnostic tests, perhaps due to lack of evidence for utility. METHODS AND RESULTS: We compared H&P estimates of filling pressures and cardiac index with invasive measurements in 194 patients in the ESCAPE trial. H&P estimates were compared with 6-month outcomes in 388 patients enrolled in ESCAPE. Measured right atrial pressure (RAP) was <8 mm Hg in 82% of patients with RAP estimated from jugular veins as <8 mm Hg, and was >12 mm Hg in 70% of patients when estimated as >12 mm Hg. From the H&P, only estimated RAP > or =12 mm Hg (odds ratio [OR] 4.6; P<0.001) and orthopnea > or =2 pillows (OR 3.6; P<0.05) were associated with pulmonary capillary wedge pressure (PCWP) > or =30 mm Hg. Estimated cardiac index did not reliably reflect measured cardiac index (P=0.09), but "cold" versus "warm" profile was associated with lower median measured cardiac index (1.75 vs. 2.0 L/min/m(2); P=0.004). In Cox regression analysis, discharge "cold" or "wet" profile conveyed a 50% increased risk of death or rehospitalization. CONCLUSIONS: In advanced HF, the presence of orthopnea and elevated jugular venous pressure are useful to detect elevated PCWP, and a global assessment of inadequate perfusion ("cold" profile) is useful to detect reduced cardiac index. Hemodynamic profiles estimated from the discharge H&P identify patients at increased risk of early events. PMID- 19675683 TI - Brief subthreshold events can act as Hebbian signals for long-term plasticity. AB - BACKGROUND: Action potentials are thought to be determinant for the induction of long-term synaptic plasticity, the cellular basis of learning and memory. However, neuronal activity does not lead systematically to an action potential but also, in many cases, to synaptic depolarizing subthreshold events. This is particularly exemplified in corticostriatal information processing. Indeed, the striatum integrates information from the whole cerebral cortex and, due to the membrane properties of striatal medium spiny neurons, cortical inputs do not systematically trigger an action potential but a wide range of subthreshold postsynaptic depolarizations. Accordingly, we have addressed the following question: does a brief subthreshold event act as a Hebbian signal and induce long term synaptic efficacy changes? METHODOLOGY/PRINCIPAL FINDINGS: Here, using perforated patch-clamp recordings on rat brain corticostriatal slices, we demonstrate, that brief (30 ms) subthreshold depolarizing events in quasi coincidence with presynaptic activity can act as Hebbian signals and are sufficient to induce long-term synaptic plasticity at corticostriatal synapses. This "subthreshold-depolarization dependent plasticity" (SDDP) induces strong, significant and bidirectional long-term synaptic efficacy changes at a very high occurrence (81%) for time intervals between pre- and postsynaptic stimulations (Deltat) of -110or=18 years attending a general practitioner in Germany. Here we describe in detail the methods used and the feasibility of determining the MetSyn in a primary health care setting. RESEARCH DESIGN AND METHODS: The German-wide cross-sectional study was performed during two weeks in October 2005. Blood samples were analyzed in a central laboratory. Waist circumference and blood pressure were assessed, data on smoking, life style, fasting status, socio-demographic characteristics and core information from non participants collected. Quality control procedures included telephone-monitoring and random on-site visits. In order to achieve a maximal number of fasting blood samples with a minimal need for follow-up appointments a stepwise approach was developed. Basic descriptive statistics were calculated, the Taylor expansion method used to estimate standard errors needed for calculation of confidence intervals for clustered observations. RESULTS: In total, 1511 randomly selected general practices from 397 out of 438 German cities and administrative districts enrolled 35,869 patients (age range: 18-99, women 61.1%). More than 50,000 blood samples were taken. Fasting blood samples were available for 49% of the participants. Of the participating patients 99.3% returned questionnaires to the GP, only 12% were not filled out completely. The overall prevalence of the MetSyn (NCEP/ATP III 2001) was found to be 19.8%, with men showing higher prevalence rates than women (22.7% respective 18.0%). CONCLUSIONS: This study was designed to provide data as robust as possible within the confines of an epidemiological study. Judging by the low degree of missing data and the high data quality, the feasibility for this kind of a research setting (short evaluation period, practitioners as data assessment sites) was found to be very good. The results will help to gain a more comprehensive insight into the prevalence of MetSyn for patients in primary health care in Germany. PMID- 19675699 TI - Why gms? PMID- 19675700 TI - Onychodystrophy and its management. AB - Onychodystrophy represents various pathologic processes of the nails such as infectious and noninfectious disorders including onychomycosis, psoriasis and allergic and irritant dermatitis. Nail changes may also be a clue to other dermatological or systemic diseases. But often it is difficult to establish the diagnosis as most nail changes are unspecific. The purpose of this article is to give an overview of the anatomy of the nail organ and possible causes of onychodystrophy. Therapeutic modalities are reviewed as well as modes of analysis. PMID- 19675701 TI - High-dose chemotherapy with autologous peripheral blood stem cell support for recurrent primary AFP-producing intracranial germinoma. AB - We report of a 34-year old man with second intracranial relapse of a suprasellar germinoma. Despite of extensive pretreatment with radiation and conventional chemotherapy relapse occurred and was treated with sequential high-dose chemotherapy followed by transfusion of autologous peripheral stem cells. The high-dose chemotherapy course was complicated by refractory derailment of pineal gland insufficiency. The patient achieved a complete remission after high dose chemotherapy which lasted for 13 months. Subsequently, he developed a third relapse and died. PMID- 19675702 TI - Effects of weight-reduction on obesity-associated diseases. AB - Even moderate, but persistent weight-loss ameliorates most of the related diseases in obesity. Besides the consequences of the metabolic syndrome, this includes less well-known obesity-associated changes, such as impaired fertility, menstrual disorders, psychic changes, total leucocyte-count as a parameter of immunity and the impaired pulmonary function in asthma and sleep-apnoea. Life expectancy is prolonged by diminution of visceral fat depots, whilst weight-loss by shrinking of fat-free body-mass seems to have a contrary effect. PMID- 19675703 TI - Disc height and anteroposterior translation in fused and adjacent segments after lumbar spine fusion. AB - In a series of 46 patients the effects of spinal fusion upon intervertebral height and sagittal alignment in operated and non-operated segments were retrospectively evaluated on digitized radiographs. Data was compared with age- and gender-normalized standard values. The objective was to evaluate the influence of different types of spine fusions primarily upon adjacent segments, particularly in terms of degeneration and sagittal profile of the lumbar spine. Incidence of adjacent segment degeneration (ASD) is still highly controversial. However, not every degeneration adjacent to spinal fusion must be caused by the fusion and responsibility of the fusion for ASD may vary with its range and type. Distortion Corrected Roentgen Analysis (DCRA) was utilized. DCRA is a proven valid, reliable, observer-independent, and accurate tool for assessment of these parameters over time and in comparison with "normal" cohorts. With this method the exact posture of the patients needs not to be known. There was little evidence for serious fusion-related ASD within an average of 40 months follow-up. No difference could be detected for rigid vs. non-rigid fusion and instrumented vs. non-instrumented techniques. Temporary postoperative distraction effects could be detected in operated and non-operated segments. Absolute preoperative values for intervertebral height and vertebral slip were age-related. Retrospectively, the choice of segments for fusion was clearly based upon radiological criteria. Thus we conclude that radiological parameters have an obvious clinical relevance for decision-making and need to be quantified. Within the limitations of this pilot study, true fusion related ASD seems to be infrequent. PMID- 19675704 TI - Prediction of treatment outcome in a clinical sample of problem drinkers: self efficacy, alcohol expectancies, and readiness to change. AB - Cognitive processes related to client motivation are important mediators of alcoholism treatment outcome. The present study aimed to expand previous research on client motivation and treatment outcome by establishing the predictive utility of self-efficacy, alcohol expectancies, and readiness to change in a sample of alcohol-dependent inpatients (N = 83). Treatment outcome was assessed three months following discharge. According to self-reported alcohol use, 22 clients were classified as abstainers and 41 clients as relapsers. Twenty participants were lost to follow-up. Readiness to change and anticipated reinforcement from alcohol predicted abstinence at follow-up. Client motivation was unrelated to both frequency and quantity of alcohol use. In accordance with social learning theory, self-efficacy was inversely correlated with alcohol expectancies. The results of the present study suggest that once abstinence has been violated factors other than pretreatment motivation determine drinking behavior. PMID- 19675705 TI - Antiviral combination therapy with interferon/peginterferon plus ribavirin for patients with chronic hepatitis C in Germany: a health technology assessment commissioned by the German Agency for Health Technology Assessment. AB - OBJECTIVE: The purpose of this health technology assessment (HTA), commissioned by the German Agency for HTA at the German Federal Ministry of Health and Social Security, was to systematically review the evidence on effectiveness and cost effectiveness of antiviral treatment (AVT) for initial chronic hepatitis C (CHC) and to apply these data in the context of the German health care system. METHODS: A systematic literature search was conducted to identify randomised controlled trials (RCTs), meta-analyses, and HTAs that evaluated initial AVT for CHC. A modified version of the German Hepatitis C Model (GEHMO)--a decision-analytic Markov model--was used to determine long-term morbidity, life expectancy, quality of life, costs and cost-effectiveness of different treatment strategies. Model parameters were derived from German databases, international RCTs, and a Cochrane Review. RESULTS: Overall, 9 RCTs, 2 HTA reports, 1 Cochrane review, and 2 meta analyses examining medical effectiveness of antiviral combination therapy, as well as 7 economic evaluations, met the inclusion criteria. These studies indicate that combination therapy with peginterferon plus ribavirin produced the highest sustained virological response rates (54-61%), followed by interferon plus ribavirin with 38-54%, and interferon monotherapy with 11-21%. Based on international cost-effectiveness studies, interferon plus ribavirin is cost effective compared to interferon monotherapy. No published articles were available regarding cost-effectiveness of peginterferon plus ribavirin. In our decision analysis, these findings were confirmed and the discounted incremental cost-effectiveness ratio for peginterferon plus ribavirin was euro 9,800 per quality-adjusted life-year gained compared to interferon monotherapy (as the next best non-dominated strategy). Sensitivity analyses showed robust results across a wide range of model parameters. CONCLUSIONS: This HTA suggests that initial combination therapy prolongs life, improves quality of life, and is cost effective in patients with CHC. Combination of peginterferon and ribavirin is the most effective and efficient treatment strategy among the examined options. PMID- 19675706 TI - PMMA vertebroplasty in patients with malignant vertebral destruction of the thoracic and lumbar spine. AB - OBJECT: Patients with osteolytic metastases frequently suffer from serious local and radicular pain. Pathophysiologically, local pain arises from skeletal instability, whereas radicular pain originates from compression of nerve roots by local tumor growth. Causal treatment of osteolytic metastases in disseminated malignant disease is very difficult. Resection of vertebrae, in combination with ventro-dorsal stabilization, is a complex treatment for patients with a limited life expectancy. Percutaneous polymethylmethacrylate (PMMA) vertebroplasty is a new and easy method of relieving patients' pain. In addition, it is both cost effective and safe. Pain is reduced immediately after treatment. Due to the regained vertebral stability, early mobilization of the patients is possible. METHODS: A total of 22 patients with osteolytic malignancies of the thoracic and lumbar spine were treated with PMMA vertebroplasty. Prior to and after surgery, then six weeks and six months after discharge from hospital, patients answered the Oswestry Low Back Pain Disability (OLBPD) Questionnaire for assessment of treatment-related change in disability. Percutaneous vertebroplasty was performed in a total of 19 patients. In three patients with tumor related compression of nerve roots an open neurolysis was performed followed by vertebroplasty. RESULTS: A total of 86% of patients reported a significant pain reduction. Vertebroplasty was highly beneficial for patients with pain related to local instability of the spine, but less so in patients with additional nerve root compression. Extravasation of PMMA beyond the vertebral margins was observed in 23% of the cases. No treatment-related clinical or neurological complications were seen. CONCLUSIONS: PMMA vertebroplasty is a useful and safe method of pain relief for patients with malignant osteolytic diseases of the thoracic and lumbar spine. PMID- 19675707 TI - Economic evaluation of newborn hearing screening: modelling costs and outcomes. AB - OBJECTIVES: The prevalence of newborn hearing disorders is 1-3 per 1,000. Crucial for later outcome are correct diagnosis and effective treatment as soon as possible. With BERA and TEOAE low-risk techniques for early detection are available. Universal screening is recommended but not realised in most European health care systems. Aim of the study was to examine the scientific evidence of newborn hearing screening and a comparison of medical outcome and costs of different programmes, differentiated by type of strategy (risk screening, universal screening, no systematical screening). METHODS: In an interdisciplinary health technology assessment project all studies on newborn hearing screening detected in a standardized comprehensive literature search were identified and data on medical outcome, costs, and cost-effectiveness extracted. A Markov model was designed to calculate cost-effectiveness ratios. RESULTS: Economic data were extracted from 20 relevant publications out of 39 publications found. In the model total costs for screening of 100,000 newborns with a time horizon of ten years were calculated: 2.0 Mio.euro for universal screening (U), 1.0 Mio.euro for risk screening (R), and 0.6 Mio.euro for no screening (N). The costs per child detected: 13,395 euro (U) respectively 6,715 euro (R), and 4,125 euro (N). At 6 months of life the following percentages of cases are detected: U 72%, R 43%, N 13%. CONCLUSIONS: A remarkable small number of economic publications mainly of low methodological quality was found. In our own model we found reasonable cost effectiveness ratios also for universal screening. Considering the outcome advantages of higher numbers of detected cases a universal newborn hearing screening is recommended. PMID- 19675708 TI - Long-term results after 110 tracheal resections. AB - OBJECTIVE: Among the many therapeutic options for treating tracheal stenosis (e.g. bouginage, laser resection and stenting), segmental resection and reconstruction with end-to-end anastomosis is the method of choice. We verified this in an analysis of clinical material. PATIENTS AND METHODS: We retrospectively evaluated 110 tracheal sleeve resections performed between 1985 and 2001. Data before and after resection were analyzed, and the patients were interviewed. RESULTS: The aetiology of stenosis was mainly postintubation injury (n = 92) (83.6%), followed by goiter with malacia (n = 8) (7.3%) and tumor (n = 6) (5.5%). There were a few other causes (n = 3) (2.7%). 48 patients (43.6%) had undergone prior conservative or surgical treatment other than sleeve resection. A cervical approach was used in 93 (84.6%), a cervicomediastinal in 15 (13.6%), and a transthoracic in two. Healing of anastomosis was uncomplicated in 101 patients (91.8%). Major and minor complications occurred in 29 patients (26.4); there were 4 dehiscences (3.6%), 3 restenoses (2.7%), 2 suture line granulations (1.8%) and 4 vocal cord dysfunctions (3.6%). The 30-day mortality rate was 0.9%. 77 patients were interviewed after surgery (median 80.1 months); 93.5% (n = 72) were satisfied with the surgical treatment. CONCLUSIONS: Resection and reconstruction offer the best treatment for tracheal stenosis. Lethal complications were due to severe comorbidity. Many patients today still undergo unsuccessful conservative treatment before being referred to surgery. PMID- 19675709 TI - Analysis of consumer information brochures on osteoporosis prevention and treatment. AB - PURPOSE: Evidence-based consumer information is a prerequisite for informed decision making. So far, there are no reports on the quality of consumer information brochures on osteoporosis. In the present study we analysed brochures on osteoporosis available in Germany. METHOD: All printed brochures from patient and consumer advocacy groups, physician and governmental organisations, health insurances, and pharmaceutical companies were initially collected in 2001, and updated in December 2004. Brochures were analysed by two independent researchers using 37 internationally proposed criteria addressing evidence-based content, risk communication, transparency of the development process, and layout and design. RESULTS: A total of 165 brochures were identified; 59 were included as they specifically targeted osteoporosis prevention and treatment. Most brochures were provided by pharmaceutical companies (n=25), followed by health insurances (n=11) and patient and consumer advocacy groups (n=11). Quality of brochures did not differ between providers. Only 1 brochure presented lifetime risk estimate; 4 mentioned natural course of osteoporosis. A balanced report on benefit versus lack of benefit was presented in 2 brochures and on benefit versus adverse effects in 8 brochures. Four brochures mentioned relative risk reduction, 1 reported absolute risk reduction through hormone replacement therapy (HRT). Out of 28 brochures accessed in 2004 10 still recommended HRT without discussing adverse effects. Transparency of the development process was limited: 25 brochures reported publication date, 26 cited author and only 1 references. In contrast, readability and design was generally good. CONCLUSION: The quality of consumer brochures on osteoporosis in Germany is utterly inadequate. They fail to give evidence-based data on diagnosis and treatment options. Therefore, the material is not useful to enhance informed consumer choice. PMID- 19675711 TI - Antioxidant vitamins and hyperbilirubinemia in neonates. AB - OBJECTIVE: Low antioxidant system may contribute to the severity of neonatal hyperbilirubinemia. The aim of this research was to explore the relationship between plasma vitamin E and C levels and the severity of hyperbilirubinemia in full-term neonates with normal glucose 6-phosphate dehydrogenase (G6PD) activities. METHODS: A total of 130 full-term healthy live birth neonates of healthy mothers with normal G6PD activity were included in this study. In addition to routine blood analysis, plasma total bilirubin, vitamin E and C levels and G6PD activity were measured on the first day of life. None of the neonates was ABO incompatible or anemic. RESULTS: Neonates who did not develop hyperbilirubinemia (n=119) had a mean plasma bilirubin level of 65+/-24 micromol/l (median 58.1), while neonates who developed significant hyperbilirubinemia (n=11) had a mean plasma bilirubin level of 238+/-56 micromol/l (median 246.2) on the first day of life. Mean plasma vitamin C levels of neonates who developed hyperbilirubinemia were significantly lower than those who did not develop hyperbilirubinemia (87+/-22 micromol/l (median 89.4) vs. 132+/-36 micromol/l (median 127.7), respectively, P=0.0001). Similar results were observed for plasma vitamin E levels in neonates who did or did not develop hyperbilirubinemia (7.5+/-2 micromol/l (median 6.3) vs. 10.4+/-5 micromol/l (median 9.1), respectively, P=0.001). Hemoglobin and hematocrit were significantly lower in neonates who developed hyperbilirubinemia (P=0.0002 and P=0.0003, respectively), although gestational age and birth weight for the two groups showed no significant difference. CONCLUSION: The results of the present work indicate that low level of plasma vitamins C and E are associated with significant hyperbilirubinemia in full-term neonates. PMID- 19675710 TI - High dose chemotherapy with autologous stem cell transplantation in diffuse large B-cell lymphoma. AB - BACKGROUND: High-dose chemotherapy (HDT) with autologous stem cell transplantation (ASCT) plays an important role in the treatment of aggressive non Hodgkin's lymphoma (NHL). We report on a retrospective analysis of all patients with diffuse large B-cell lymphoma who were consecutively treated with HDT followed by ASCT at the University Hospital of Bonn, Germany, between 1996 and 2004. METHODS: A total of 25 patients were transplanted for biopsy-proven diffuse large B-cell lymphoma (DLBCL). Eight patients received up-front HDT as first-line therapy, four patients received HDT due to incomplete response to conventional induction chemotherapy, and six patients were treated for primary refractory disease. Seven patients had recurrent lymphoma. RESULTS: A complete remission (CR) was achieved in 14 of 25 patients (56%). Estimated 3-year survival for patients treated with upfront HDT, chemosensitive patients with incomplete response to first line therapy, and patients with chemosensitive relapsed disease was 87.5%, 50.0% and 60.0%, respectively. In contrast, no patient with primary refractory disease or relapsed disease lacking chemosensitivity lived longer than 8 months. Chemosensitivity was the only significant prognostic factor for overall survival (OS) in multivariate analysis. CONCLUSIONS: Our results confirm that HDT and ASCT is a highly effective therapy in patients with DLBCL leading to long term survival in a substantial proportion of patients. Patients treated upfront for high-risk disease, incomplete response to conventional first-line therapy, or for chemosensitive relapse have a good prognosis. In contrast, patients with primary chemorefractory disease and patients with relapsed disease lacking chemosensitivity do not benefit from HDT with ASCT. PMID- 19675712 TI - Atherosclerosis, cholesterol, nutrition, and statins--a critical review. AB - Atherosclerosis, which causes approximately half of all deaths of adults over age 60 in industrialized nations, is a pandemic among inappropriately nourished and/or physically hypoactive children, adolescents, and adults world wide. Although nowadays statins are widely prescribed to middle age and elderly adults with high blood lipid levels as pharmacological prevention for the late complications of atherosclerosis, from a critical point of view statins seem not to solve the problem, especially when compared with certain natural ingredients of our nutrition like micronutrients as alternative strategy. Statin ingestion is associated with lowering of serum cholesterol and low-density lipoprotein concentrations; some prospective studies have shown statistical associations with subsequent modest reduction of mortality from cardiovascular disease. However, specific biochemical pathways and pharmacological roles of statins in prevention of atherosclerosis, if any, are unknown. Moreover, there have been no systematic cost-benefit analyses of life-style prophylaxis versus statin prophylaxis versus combined life-style plus statin prophylaxis versus neither life-style nor statin prophylaxis for clinically significant complications of cardiovascular diseases in the elderly. Further, in the trials of effectiveness statins were not compared with management of nutrition, which is the most appropriate alternative intervention. Such studies seem to be important, as the ever increasing world population, especially in developing countries, now demand expensive statins, which may be unaffordable for mitigating the pandemic. Studies of this kind are necessary to identify more precisely those patients for whom cardiovascular benefits will outweigh the risks and costs of the statin treatment in comparison with nutritional interventions. Against the background of the current pathogenetic concept of atherogenesis some of its possible risk factors, particularly the roles of cholesterol and homocysteine, and the effects of statins versus nutritional (micronutrients) interventions in prevention and treatment of the disease are discussed. The prevailing opinion that serum cholesterol as a mediator of the disease is increased by eating saturated fats and decreased by eating polyunsaturated fats is being challenged. Evidently, the beneficial effects of statins in atherosclerosis are not mainly due to its cholesterol lowering effect, rather than to its "pleiotropic effects". Other pathogenetic factors in atherosclerosis are involved, like inflammatory and immunologic processes, that can be modulated by statins as well as by other drugs or by the Mediterranean-style nutrition and by micronutrients (folate, B vitamins). PMID- 19675713 TI - Selective information seeking: can consumers' avoidance of evidence-based information on colorectal cancer screening be explained by the theory of cognitive dissonance? AB - BACKGROUND: Evidence-based patient information (EBPI) is a prerequisite for informed decision-making. However, presentation of EBPI may lead to irrational reactions causing avoidance, minimisation and devaluation of the information. OBJECTIVE: To explore whether the theory of cognitive dissonance is applicable to medical decision-making and useful to explain these phenomena. SETTING AND PARTICIPANTS: 261 volunteers from Hamburg (157 women), >or=50 years old without diagnosis of colorectal cancer. DESIGN AND VARIABLES: Within an experiment we simulated information seeking on colorectal cancer screening. Consumers' attitudes towards screening were surveyed using a rating scale from -5 (participate in no way) to +5 (participate unconditionally) (independent variable). Using a cover story, participants were asked to sort 5 article headlines according to their reading preferences. The headlines simulated the pro to contra variety of contents to be found in print media about colorectal cancer screening. The dependent variable was the sequence of article headlines. RESULTS: Participants were very much in favour of screening with scores for faecal occult blood test of 4.0 (0.1) and for colonoscopy 3.3 (0.1). According to our hypothesis we found statistically significant positive correlations between the stimuli in favour of screening and attitudes and significant negative correlations between the stimuli against screening and attitudes. CONCLUSION: The theory of cognitive dissonance is applicable to medical decision-making. It may explain some phenomena of irrational reactions to evidence-based patient information. PMID- 19675714 TI - Central venous port system associated thromboses: outcome in 3498 implantations and literature review. AB - METHODS: From 1 July 1995 to 31 June 2006 we implanted 3498 intravenous port systems. In nearly all cases the indication was vascular access for chemotherapy. RESULTS: We registered 199 complications (5.7%), mostly infections (n=85 i.e. 2.4%) and thromboses (n=63 i.e. 1.8%). CONCLUSIONS: Permanent central venous catheters have become standard in the management of patients with malignancies. Because of the improvement of material and design during the past twenty years technical complications have been reduced significantly. The most frequent occurring medical complications are infection and thromboses. In order to further minimize these disadvantages we developed a "best practices" standard for port implantation combining own data with recent studies. PMID- 19675715 TI - Cytokine-induced killer cells are type II natural killer T cells. AB - BACKGROUND: Until now, cytokine-induced killer (CIK) cells were assumed to be part of the type I natural killer T (NKT) cell population, but it was not yet investigated if this is correct. METHODS: For analysis, CIK cells were generated by various culture conditions. Human type I NKT cells express a T cell receptor (TCR) composed of an invariant V alpha 24-J alpha Q chain combined with one of several V beta chains. The V alpha 24 is a reliable marker for the presence of these TCRs. RESULTS: While comparing cultures stimulated with different substances, we observed the lack of any V alpha 24 on the surface of CIK culture cells. CONCLUSION: We conclude that CIK cells do not belong to the type I NKT cells. PMID- 19675717 TI - Excessive working hours and health complaints among hospital physicians: a study based on a national sample of hospital physicians in Germany. AB - OBJECTIVES: To determine correlations between excessively long working hours and subjectively experienced somatic health complaints among hospital physicians. METHODS: Quantitative data were collected as part of the survey "Working life, Lifestyle and Health of Hospital Physicians in Germany 2006" using self-reporting questionnaires. The individually experienced health was assessed on the basis of Zerssen's list of somatic complaints. The indicator of excessively long working hours was defined as 10 or more working hours per working day and 6 or more on call shifts a month among full-time employees. The net sample consisted of 3295 randomly selected physicians from 515 hospitals. RESULTS: The response rate was 58% (n=1917). Physicians with excessively long working hours (19%) had significantly higher sum score of health complaints (p=0.0001) and significantly increased mental and physical fatigue symptoms (feeling faint, languor, uneasiness, heavy legs, excessive need for sleep, trembling; p=0.0001 to 0.047), mood changes (irritability, brooding; p=0.008 to 0.014), gastrointestinal (nausea, loss of weight; p=0.0001 to 0.014) and heart disorders (lumpy sensation in the throat, chest pain; p=0.0001 to 0.042). When the sum score of health complaints was controlled for selected confounders, being female (B=-3.44, p=0.0001) and having excessively long working hours (B=2.76, p=0.0001) were significantly correlated with health complaints. In a separate gender analysis, being exposed to excessively long working hours remained a significant predictor for health complaints among both females (B=3.78, p=0.001) and males (B=2.28, p=0.004). CONCLUSIONS: Excessively long working hours are associated with an increased risk of health complaints. Reducing working hours may be the first step to improving physicians' health. PMID- 19675716 TI - Water-filtered infrared-A radiation (wIRA) is not implicated in cellular degeneration of human skin. AB - BACKGROUND: Excessive exposure to solar ultraviolet radiation is involved in the complex biologic process of cutaneous aging. Wavelengths in the ultraviolet-A and -B range (UV-A and UV-B) have been shown to be responsible for the induction of proteases, e. g. the collagenase matrix metalloproteinase 1 (MMP-1), which are related to cell aging. As devices emitting longer wavelengths are widely used in therapeutic and cosmetic interventions and as the induction of MMP-1 by water filtered infrared-A (wIRA) had been discussed, it was of interest to assess effects of wIRA on the cellular and molecular level known to be possibly involved in cutaneous degeneration. OBJECTIVES: Investigation of the biological implications of widely used water-filtered infrared-A (wIRA) radiators for clinical use on human skin fibroblasts assessed by MMP-1 gene expression (MMP-1 messenger ribonucleic acid (mRNA) expression). METHODS: Human skin fibroblasts were irradiated with approximately 88% wIRA (780-1400 nm) and 12% red light (RL, 665-780 nm) with 380 mW/cm(2) wIRA(+RL) (333 mW/cm(2) wIRA) on the one hand and for comparison with UV-A (330-400 nm, mainly UV-A1) and a small amount of blue light (BL, 400-450 nm) with 28 mW/cm(2) UV-A(+BL) on the other hand. Survival curves were established by colony forming ability after single exposures between 15 minutes and 8 hours to wIRA(+RL) (340-10880 J/cm(2) wIRA(+RL), 300-9600 J/cm(2) wIRA) or 15-45 minutes to UV-A(+BL) (25-75 J/cm(2) UV-A(+BL)). Both conventional Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) and quantitative real-time RT-PCR techniques were used to determine the induction of MMP-1 mRNA at two physiologic temperatures for skin fibroblasts (30 degrees C and 37 degrees C) in single exposure regimens (15-60 minutes wIRA(+RL), 340-1360 J/cm(2) wIRA(+RL), 300-1200 J/cm(2) wIRA; 30 minutes UV-A(+BL), 50 J/cm(2) UV A(+BL)) and in addition at 30 degrees C in a repeated exposure protocol (up to 10 times 15 minutes wIRA(+RL) with 340 J/cm(2) wIRA(+RL), 300 J/cm(2) wIRA at each time). RESULTS: Single exposure of cultured human dermal fibroblasts to UV-A(+BL) radiation yielded a very high increase in MMP-1 mRNA expression (11 +/-1 fold expression for RT-PCR and 76 +/-2 fold expression for real-time RT-PCR both at 30 degrees C, 75 +/-1 fold expression for real-time RT-PCR at 37 degrees C) and a dose-dependent decrease in cell survival. In contrast, wIRA(+RL) did not produce cell death and did not induce a systematic increase in MMP-1 mRNA expression (less than twofold expression, within the laboratory range of fluctuation) detectable with the sensitive methods applied. Additionally, repeated exposure of human skin fibroblasts to wIRA(+RL) did not induce MMP-1 mRNA expression systematically (less than twofold expression by up to 10 consecutive wIRA(+RL) exposures and analysis with real-time RT-PCR). CONCLUSIONS: wIRA(+RL) even at the investigated disproportionally high irradiances does not induce cell death or a systematic increase of MMP-1 mRNA expression, both of which can be easily induced by UV-A radiation. Furthermore, these results support previous findings of in vivo investigations on collagenase induction by UV-A but not wIRA and show that infrared-A with appropriate irradiances does not seem to be involved in MMP-1 mediated photoaging of the skin. As suggested by previously published studies wIRA could even be implicated in a protective manner. PMID- 19675718 TI - Apparent contradiction between negative effects of UV radiation and positive effects of sun exposure. PMID- 19675719 TI - External quality assessment of tumour marker analysis: state of the art and consequences for estimating diagnostic sensitivity and specificity. AB - This review shows the current analytical quality for the following analytes used as tumour markers in the external quality assessment (EQA)-programmes of Instand e.V., a national EQA-organiser in Germany: Corticotropin (ACTH), growth hormone (GH, hGH), prolactin (PRL), chorionic gonadotropin (CG, hCG), calcitonin (CT, hCT), thyroglobulin (Tg), carcinoembryonic antigen (CEA), CA-Antigens 125, 72-4, 15-3 and 19-9, alpha foetoprotein (AFP) and prostate-specific antigen (PSA). The results from the participants show a large variation in the precision of the methods used as well as in the comparability of results between methods for the same analyte. In general, the hormones used as tumour markers show better performance than the "CA-markers", which are often inadequately standardised and defined. In the case of one CA-marker (CA 72-4/TAG 72-4), the differences between the lowest kit median concentration and highest kit median concentration for one sample pair were 440% and 580%. The corresponding figures for ACTH were 123% and 156% and for CEA 180% and 184%. The classical tumour markers such as carcinoembryonic antigen (CEA) and alpha foetoprotein (AFP) performed markedly better than the CA-markers and PSA with regards to both inter- and intra-method comparability. The inter-laboratory precision for a given kit and marker was acceptable in many cases. The results show that only results from the same kit/method for each tumour marker can be used for cumulative or time-dependent comparison of results - for example pre-operative and post-operative follow up. In the case of prostate specific antigen (PSA), the kits used for free and total PSA must come from the same producer, if the generally accepted ratios are to have any diagnostic value. The need for kit- and laboratory-specific reference ranges and cut-off values for setting diagnostic specificity and sensitivity is highlighted from the EQA-results. The situation for inter-method comparability for the CA-Markers has not improved over the past decade. With the exception of calcitonin for detecting medullary thyroid carcinoma, chorionic gonadotropin in germ-cell tumours in men and thyroglobulin after total thyroidectomy, none of the remaining analytes appear to be suitable for screening purposes. PMID- 19675720 TI - Acute lung affection in an endurance-trained man under amiodarone medication. AB - Patients undergoing treatment with amiodarone can develop severe pulmonary side effects. This effect, which is often highly underestimated, can lead to dyspnea, pneumonitis, and further fibrosis. A recent change in the labeling of amdiodarone by the American Food and Drug Administration (FDA) supports this suspicion. Tracing the symptoms back to the causing agent can be difficult, as shown in our report. The subject of this case report is an endurance-trained 65 year old male marathon runner who appeared with atrial fibrillation during a routine check up in autumn 2003. After medical cardioversion with flecainide a complaint free interval of 8 months was followed by a relapse, which resulted in a change of medication to amiodarone. Due to misunderstandings the patient kept on taking the amiodarone loading dose for six weeks and returned with severe dyspnea on exertion. Losses in CO diffusing capacity, a lowered macrophages count and a positive lymphocyte transformation test were the only first hand clinical evidence of amiodarone intoxication, despite the sensation of dyspnea. This case shows that special care has to be taken in treatment with amiodarone. Side effects can be hard to trace and do not evidently show a clear connection to amiodarone. PMID- 19675721 TI - Beta-trace protein (prostaglandin D synthase)--a stable and reliable protein in perilymph. AB - OBJECTIVE: Beta-trace protein (beta-TP) has been analysed in human cerebrospinal fluid (CSF) and other body fluids. Beta-trace protein is a very sensitive and specific clinical marker and can confirm reliably the presence of CSF in patients with a suspected CSF leakage. DESIGN: Perilymph specimens from the scala vestibuli (n=10) and from the lateral semicircular canal (n=4) were taken from patients undergoing stapedotomy or surgery for acoustic neuroma. During post mortem examinations perilymph specimens from the scala vestibuli (n=70), the scala tympani (n=11), endolymph specimens (n=21) and CSF specimens (n=17) were obtained. All specimens were analyzed by a one-dimensional immunoelectrophoresis using a polyclonal, monospecific antibody. RESULTS: Specimens from live surgery showed a mean concentration of 51.5 +/- 48.9 mg/l beta-TP in scala vestibuli perilymph. Specimens from post-mortem examinations revealed a mean concentration of 49.1 +/- 17.7 mg/l in CSF, 71.9 +/- 29.3 mg/l in perilymph and 68.0 +/- 21.7 mg/l in endolymph. There was no evidence of a circadian alteration of beta-TP in CSF or inner ear fluids. CONCLUSIONS: Our results demonstrated clearly that beta TP is contained in human perilymph and endolymph. This is the first published data that point out the aptitude of the beta-TP-test in verifying traces of perilymph, a valuable diagnostic tool for the existence of perilymphatic leaks. PMID- 19675722 TI - Hoffmann's syndrome: a case report. AB - This syndrome is characterized by the presence of hypothyroidism with myxoedema, muscle stiffness and pseudo hypertrophy. We describe the disorder in a 21 year old male, who got admitted with complaints of generalized weakness, cold intolerance, constipation, and hoarse voice, difficulty in walking and progressive enlargement of muscles of thighs and back with crampy pains for two years. Examination revealed mild mental retardation, enlarged tongue, dry and rough skin, enlargement of thighs and back muscles, motor weakness in flexors of hips and knees with delayed relaxation of deep tendon reflexes. Investigations revealed evidence of hypothyroidism with marked elevation of muscle enzymes. Following institution of replacement therapy with thyroxine, the patient showed marked clinical and biochemical improvements after six months, but insignificant decrease in muscle mass. In this report we review relevant literature. PMID- 19675724 TI - Prevalence of problem-related drinking among doctors: a review on representative samples. AB - AIMS: This paper is a review of the literature on problem-related drinking of alcohol among medical doctors, and it deals with the epidemiology and results. METHODS: A search of computer literature databases - PubMed and ETOH - was performed to locate articles reporting problem-related drinking among doctors, using population-based samples of doctors within the last two decades. RESULTS: In the light of different definitions of problem-related drinking, there was found a breadth of prevalence of problem-related drinking - from heavy drinking and hazardous drinking (12%-16%) to misuse and dependence (6%-8%) - within the population-based samples of doctors. An increased risk was positively related to male doctors and doctors of the age of 40-45 years and older, and to some factors of work, lifestyle and health. CONCLUSION: For the future, it seems necessary to sensitise the research for problem-related drinking of doctors in Germany, e.g. initiating a representative survey, analysing the drinking of alcohol in the context of health, life-style and work-related factors. PMID- 19675723 TI - Surveillance of nosocomial infections at a Saudi Arabian military hospital for a one-year period. AB - The objectives of the current study are to define how many and what kind of nosocomial infections are occurring, what are the causative microbes and what kind of drugs can be used in treatment of infection at Al-Hada Armed Forces Hospital, Taif, Saudi Arabia during the year 2004. A prospective study was implemented for all cases admitted at Al-Hada Armed Forces Hospital during the period 1(st) January, 2004 till 31(st) December, 2004 and which developed infection. Determination of nosocomial infections was performed using standardized CDC criteria. A total of 1382 patients had developed infection during hospital admission and were included in the study. Of them, 668 (48.3%) had nosocomial infection and 714 (51.7%) had community-acquired infection. Among those who developed nosocomial infections, 216 (32.3%), 172 (25.7%) and 124 (18.6%) had respiratory tract (RTI), urinary tract (UTI) and blood stream infections (BSI) respectively. Surgical site infection (SSI) was reported in 86 cases (12.9%). The overall nosocomial infection rate along the study period was 4.98 per 100 discharged patients. Gram-positive organisms were reported in 31.8%. MRSA (Methicillin-resistant S. aureus) was the commonest (10.2%), followed by coagulase negative staphylococci (8.5%) and MSSA (Methicillin-susceptible S. aureus, 7.4%). While Gram-negative organisms were reported in 66.2%, E. coli was the commonest (22.3%), followed by Pseudomonas aeruginosa (17.6%) and Klebsiella pneumoniae (9.9%). Acinetobacter spp. and MRSA were highly sensitive to Imipenem (88.6%) and Vancomycin (98.5%) respectively. E. coli were highly sensitive to most of the antimicrobial agents except ampicillin (26.6%). CONCLUSIONS: Pneumonia, urinary tract infections, and blood stream infections made up the great majority of nosocomial infections. There is a need for further risk assessment associated with main types of infection. PMID- 19675726 TI - Experimental treatment of stroke in spontaneously hypertensive rats by CD34+ and CD34- cord blood cells. AB - Human umbilical cord blood as a source of stem cells has recently been reported in experimental treatment of cerebral disorders. However, little is known about the nature of cells and cellular mechanisms leading to neurofunctional improvement. Here we investigated the potential of separated CD34(+) versus CD34( ) human umbilical cord blood cells (HUCBC) to promote functional recovery following stroke. The experiments were performed in spontaneously hypertensive (SH) rats, known for a risk profile comparable to stroke patients. After three weeks of behavioral training in the RotaRod and Beamwalk test arrays, stroke was induced by permanent middle cerebral artery occlusion (MCAO). For cell therapy, 1 x 10(6) cryopreserved cells were administered systemically between 8 and 10 hours after MCAO. The behavioral tests were performed together with a neurological severity score (mNSS) until day 29 to assess neurofunctional disabilities. Nearly complete functional remission was observed with both subpopulations CD34(+) as well as CD34(-) cells. To localize cells histologically, they were labeled with a fluorescence dye (CFSE) before injection. Again, after administration of CD34(+) as well as CD34(-) cells, CFSE labelled cells were found that accumulated in the border zone between the central necrosis of the ischemic lesion and functional brain tissue, thus indicating active attraction towards the lesion for both cell populations. Immunohistology with anti-CD68 and antibodies to human neuronal markers (NF-L, chromogranin) indicated an accumulation of human and rat monocytes in the border zone of the lesion while neuronal cells of human origin could not be detected in host brains. PMID- 19675727 TI - A new bipolar blood sealing system embedded in perioperative strategies vs. a conventional regimen for total knee arthroplasty: results of a matched-pair study. AB - From a prospective matched-pair study in 40 patients it can be concluded that the use of a new bipolar sealer system for electrocoagulation is effective in Total Knee Arthroplasty in reducing the visible total blood loss until drain removal on day two after surgery by more than 28% as compared to a conventional regimen. Together with shed blood autotransfusion, it reduced homologous blood transfusions by a factor of five. No complications occurred. The system can be recommended for use in knee surgery during, after opening, and without use of a tourniquet. It bears additional potential for further applications in orthopaedic surgery. PMID- 19675725 TI - State of the art of neoadjuvant chemotherapy in breast cancer: rationale, results and recent developments. AB - Aims, results, advantages and possible disadvantages of preoperative chemotherapy (pCHT) for breast cancer are discussed in this review. Established chemotherapeutic regimens are described with respect to new drugs that are added to combinations now and in the future. Illustrating the potential of new components, trastuzumab and cytotoxic chemotherapy, were combined in neoadjuvant trials for the first time. This approach yielded impressing and unprecedented high pathological response rates. An overview regarding current neoadjuvant cytostatic and immunotherapy trials is given. Established prognostic factors like axillary lymph-nodal status are altered during pCHT, which causes the need for new prognostic markers. The consequences of these changes for clinical decision making are demonstrated. It seems possible that the advances of gene array and protein expression profile technologies will lead to improved prognostic and predictive statements. Tumor tissue can be analyzed before during and after treatment in this regard recent studies investigating the response to specific, chemotherapeutics in correlation to molecular markers are reviewed. These approaches might enable us to identify chemoresistance of specific tumors. Furthermore pCHT allows testing of chemosensitivity in vivo in an early stage, which might lead to a more individualized cancer therapy. We discuss radiotherapy after neoadjuvant therapy and the risk of local relapse after breast conserving surgery, which was made feasible by pCHT. It is shown how the evaluation of efficacy of new cancer drugs, using the neoadjuvant situation, can be done more rapidly than in the metastatic and adjuvant setting. PMID- 19675728 TI - Is the aptitude of manual skills enough for assessing the training effect of students using a laparoscopy simulator? AB - BACKGROUND: The aim of this study was to determine if students are suitable candidates to assess the learning effect through a virtual reality laparoscopy simulator (LapSim). MATERIALS AND METHODS: 14 medical students in their final year without any previous experience with a virtual reality simulator were recruited as subjects. In order to establish a "base line" all subjects were instructed into the "clip application" task - a basis module of the laparoscopy simulator - at the beginning of the study. They were then randomized into two groups. Group A (n=7) had parameter adjusted to an easy level of performance, while group B (n=7) was adjusted to a difficult level. In both levels, errors simulated clinically relevant situations such as vessel rupture and subsequent bleeding. Each participant had to repeat the clip application task ten times consecutively. RESULTS: The mean time for completion ten repetitions was 15 min pro participant in group A and 20 min in group B. From the first to the fifth repetition group A improved significantly the task completion time from 238.9 s to 103.3 s (p<0.007) consecutively and also improved the error score from 312 to 177 (p<0.07). At the tenth repetition they increased the task completion time from 103.3 s to 152.2 s (p<0.09) and increased their error score from 177 to 202 (p=0.25).From the first to the fifth repetition group B also improved the task completion time from 131.6 s to 104.5 s (p<0.31) consecutively and improved the error score from 235 to 208 (p<0.32) but at the tenth repetition they increased the task completion time from 104.5 s to 142.4 s (p<0.45) and clearly increased their error score from 208 to 244 (p<0.38). CONCLUSION: These results suggest that medical students, who lack clinical background, may be not suitable candidates for assessing the efficiency of a training model using a laparoscopy simulator. If medical students are appointed for such studies, they should receive didactic sessions in the context of a clinical curriculum prior to manual training. PMID- 19675729 TI - Children in clinical trials: survey on the current situation in paediatric university clinics in Germany. AB - Many prescribed treatments for children have not been adequately tested in children, sometimes resulting in harmful treatments being given and beneficial treatments being withheld. In the absence of specific trial-based data in children, results of studies in adults are extrapolated, which is often inappropriate because children have different range of diseases and metabolize medications differently. Trials in children are more challenging than those in adults and the pool of eligible children entering trials is often small. Children must have at last the same rights as adults in relation to receiving treatment with medicinal products that have been fully tested. The need for more studies to obtain paediatric information for medicines used in children is now a matter of consensus on a global basis and is considered a public health priority. Therefore a survey was performed in university hospitals in Germany targeting the current and future situation of children in clinical trials. The questionnaire of this survey was sent to 68 paediatric departments in 31 university clinics in Germany with a respond rate of 27% with respect to 18 returned questionnaires. With regard to new laws, guidelines and strong governmental support and funding an increasing number of clinical trials is expected. Surprisingly, the number of trials in the paediatric population remains unchanged within a period of 4 years (2005-2008). Added to the surveys performed within the pharmaceutical industry from Heinrich and Hark the number of trials in children remains unchanged even within a period of 9 years (2000-2008). The efforts undertaken by the government regarding funding and supporting KKS (Coordinating Centers for Clinical Trials) and affiliated PAED-Net (Pediatric Network on Medication Development and Testing in Children and Adolescents at KKS) appear to be insufficient. Beginning of this year the legal framework with the urgent expected "Paediatric Regulation" was established. May be the implementation by clinicians and pharmaceutical industry will improve the current situation. PMID- 19675730 TI - A note on the use of the non-parametric Wilcoxon-Mann-Whitney test in the analysis of medical studies. AB - BACKGROUND: Although non-normal data are widespread in biomedical research, parametric tests unnecessarily predominate in statistical analyses. METHODS: We surveyed five biomedical journals and - for all studies which contain at least the unpaired t-test or the non-parametric Wilcoxon-Mann-Whitney test - investigated the relationship between the choice of a statistical test and other variables such as type of journal, sample size, randomization, sponsoring etc. RESULTS: The non-parametric Wilcoxon-Mann-Whitney was used in 30% of the studies. In a multivariable logistic regression the type of journal, the test object, the scale of measurement and the statistical software were significant. The non parametric test was more common in case of non-continuous data, in high-impact journals, in studies in humans, and when the statistical software is specified, in particular when SPSS was used. PMID- 19675731 TI - "Don't know" answers concerning somatic disease status should not be regarded as "no" responses. AB - BACKGROUND: With regard to patients' self-reported somatic diseases some researchers transformed "don't know (DK)" responses into "no" responses. The present study examines the appropriateness of this procedure. METHODS: Analyses were based on the nationally representative German National Health Interview and Examination Survey (GHS), which assessed both self-reported diseases and physician-diagnosed diseases (N = 7124). Prevalence rates of persons' DK responses and the corresponding prevalences of physicians' diagnoses were calculated for persons with hypertension, coronary heart disease (CHD), heart failure, asthma, chronic bronchitis, thyroid disease, diabetes, cancer, gout, arthrosis, arthritis and osteoporosis. Correlates of physicians' diagnosed diseases of DK cases are reported. RESULTS: Between 1.6% and 9.8% of the participants responded with DK to the question of whether they have the disease. In 3.7% to 29.5% of DK cases, the physicians did regard the respective disease as being present. With regard to persons who responded with DK, the probability of a physicians' diagnosis was increased in the case of increased age and a higher number of somatic comorbidities. CONCLUSION: The procedure of transforming DK responses into "no" answers does not appear to be recommendable. PMID- 19675732 TI - Subacute transverse myelitis with Lyme profile dissociation. AB - INTRODUCTION: Transverse myelitis is a very rare neurologic syndrome with an incidence per year of 1-5 per million population. We are presenting an interesting case of subacute transverse myelitis with its MRI (magnetic resonance imaging) and CSF (cerebrospinal fluid) findings. CASE: A 46-year-old African American woman presented with decreased sensation in the lower extremities which started three weeks ago when she had a 36-hour episode of sore throat. She reported numbness up to the level just below the breasts. Lyme disease antibodies total IgG (immunoglobulin G) and IgM (immunoglobulin M) in the blood was positive. Antinuclear antibody profile was within normal limits. MRI of the cervical spine showed swelling in the lower cervical cord with contrast enhancement. Cerebrospinal fluid was clear with negative Borrelia Burgdorferi IgG and IgM. Herpes simplex, mycoplasma, coxiella, anaplasma, cryptococcus and hepatitis B were all negative. No oligoclonal bands were detected. Quick improvement ensued after she was given IV Ceftriaxone for 7 days. The patient was discharged on the 8(th) day in stable condition. She continued on doxycycline for 21 days. CONCLUSIONS: Transverse myelitis should be included in the differential diagnosis of any patient presenting with acute or subacute myelopathy in association with localized contrast enhancement in the spinal cord especially if flu-like prodromal symptoms were reported. Lyme disease serology is indicated in patients with neurological symptoms keeping in mind that dissociation in Lyme antibody titers between the blood and the CSF is possible. PMID- 19675733 TI - Modulation of formalin-induced fos-like immunoreactivity in the spinal cord by swim stress-induced analgesia, morphine and ketamine. AB - Induction of c-fos in the spinal cord due to pain is well established. This study aims to look at the effects of acute swim stress on Fos-like immunoreactivity (FLI) induced by formalin and how it is modulated by ketamine and morphine. Acutely-stressed and non-stressed adult male Sprague Dawley rats were pretreated with intraperitoneal injection of ketamine 5 mg/kg (Ketava, Atlantic Lab), morphine 10 mg/kg (Rhotard, Custom Pharmaceutical), or saline, 5 minutes prior to experimentation. Rats were acutely stressed by swimming for 3 min in 20 degrees C water. Dilute formalin (Formaldehyde, Merck) was injected to the hindpaw and the formalin score recorded. Rats were then sacrificed and spinal cords (L4-L5) removed for immunohistochemical analysis of FLI. Two-way ANOVA showed significant effects of stress, drug and stress-drug interactions in formalin test and FLI. Both morphine and ketamine produced analgesia in the formalin test. In the saline stressed group, FLI was suppressed on the ipsilateral side (p<0.01) but increased on the contralateral side (p<0.01) compared with non-stressed saline. In morphine and ketamine stressed groups, FLI was increased on both ipsilateral and contralateral sides for morphine (ipsilateral: p<0.05; contralateral: p<0.001) and ketamine (ipsilateral: p<0.05, contralateral: p<0.05) compared with their corresponding non-stressed groups. In conclusion, presence of stress may lead to discrepancy between behavioural manifestation of pain and c-fos induction in the spinal cord. PMID- 19675734 TI - Occupational skin diseases: options for multidisciplinary networking in preventive medicine. AB - Occupational dermatoses (OD) have topped the list of occupational diseases in Germany for years. Presently, approximately 16,000 new OD cases are officially reported to public statutory employers' liability insurance bodies, each year. The disease burden is high not only for individuals but also for society as a whole. Estimated annual economic costs in Germany due to sick-leave and lack of productivity due to OD are more than 1.5 billion euros. Thus, in recent years, various pilot initiatives aiming to improve prevention of occupational skin diseases (of various degrees of severity) have been developed and recently evaluated in Osnabruck. These activities have been funded by statutory employers' liability insurance schemes. Concepts underpinning these initiatives include multidisciplinary skin protection teaching programs for various high-risk professions, which turned out to be pivotal for the success of these projects. A corollary of this work is a nationwide multi-step intervention approach currently implemented by the public statutory insurance system. This approach offers quick preventive help for all levels of severity of OD. These nation-wide activities are accompanied by a national Prevention Campaign: Skin 2007/2008 (Figure 1 (Fig. 1)), which focuses mainly on primary prevention. Despite the high prevalence of OD and its poor prognosis, little is known about the molecular mechanisms underlying individual susceptibility to develop chronic irritant dermatitis. Skin irritation tests are thus far of only limited value. Presently, our institution, in collaboration with Amsterdam universities, focuses on immunogenetic risk factors potentially involved in individual susceptibility to OD in order to improve pre-employment counseling and predictive skin testing. For early secondary prevention, the so-called dermatologist's procedure was recently up dated in order to provide more rapid dermatological consultation. Additionally, combined outpatient dermatological and educational intervention seminars (secondary individual prevention, SIP) are offered to affected employees. We recently demonstrated the sustainability of the SIP approach in hairdressing for periods of up to 10 years. For those cases of OD, in which the abovementioned outpatient prevention measures are not sufficiently successful, specific interdisciplinary inpatient prevention measures have been developed (tertiary individual prevention, or TIP). TIP represents the ultima ratio within the hierarchical prevention concept of the Osnabruck Model. TIP comprises 2-3 weeks of inpatient dermatological diagnostics and treatment as well as intensive health related pedagogic and psychological counseling. Subsequent to this, 3 consecutive weeks of outpatient treatment are given by a local dermatologist. Each patient remains on sick-leave for a total of 6 weeks to allow full barrier recovery. A total of 764 out of 1164 (66%) TIP patients treated in our university, followed up regularly by a local dermatologist for up to 1 year, were successful in remaining in their respective (risk-) professions as assessed by questionnaire 1 year after discharge. Recently obtained SIP and TIP data reveal that there are reliable, evidence-based options for multidisciplinary prevention and patient management of OD, using a combined approach by a network of clinics, practices and statutory social insurance bodies. A multicentre study, which aims to further standardize TIP and evaluate sustainability of prevention in more depth (3-year dermatological follow-up of 1000 OD patients) is currently being conducted in Germany. PMID- 19675735 TI - Water-filtered infrared-A (wIRA) can act as a penetration enhancer for topically applied substances. AB - BACKGROUND: Water-filtered infrared-A (wIRA) irradiation has been shown to enhance penetration of clinically used topically applied substances in humans through investigation of functional effects of penetrated substances like vasoconstriction by cortisone. AIM OF THE STUDY: Investigation of the influence of wIRA irradiation on the dermatopharmacokinetics of topically applied substances by use of optical methods, especially to localize penetrating substances, in a prospective randomised controlled study in humans. METHODS: The penetration profiles of the hydrophilic dye fluorescein and the lipophilic dye curcumin in separate standard water-in-oil emulsions were determined on the inner forearm of test persons by tape stripping in combination with spectroscopic measurements. Additionally, the penetration was investigated in vivo by laser scanning microscopy. Transepidermal water loss, hydration of the epidermis, and surface temperature were determined. Three different procedures (modes A, B, C) were used in a randomised order on three separate days of investigation in each of 12 test persons. In mode A, the two dyes were applied on different skin areas without water-filtered infrared-A (wIRA) irradiation. In mode B, the skin surface was irradiated with wIRA over 30 min before application of the two dyes (Hydrosun radiator type 501, 10 mm water cuvette, orange filter OG590, water-filtered spectrum: 590-1400 nm with dominant amount of wIRA). In mode C, the two dyes were applied and immediately afterwards the skin was irradiated with wIRA over 30 min. In all modes, tape stripping started 30 min after application of the formulations. Main variable of interest was the ratio of the amount of the dye in the deeper (second) 10% of the stratum corneum to the amount of the dye in the upper 10% of the stratum corneum. RESULTS: The penetration profiles of the hydrophilic fluorescein showed in case of pretreatment or treatment with wIRA (modes B and C) an increased penetration depth compared to the non-irradiated skin (mode A): The ratio of the amount of the dye in the deeper (second) 10% of the stratum corneum to the amount of the dye in the upper 10% of the stratum corneum showed medians and interquartile ranges for mode A of 0.017 (0.007/0.050), for mode B of 0.084 (0.021/0.106), for mode C of 0.104 (0.069/0.192) (difference between modes: p=0.0112, significant; comparison mode A with mode C: p<0.01, significant). In contrast to fluorescein, the lipophilic curcumin showed no differences in the penetration kinetics, in reference to whether the skin was irradiated with wIRA or not. These effects were confirmed by laser scanning microscopy. Water-filtered infrared-A irradiation increased the hydration of the stratum corneum: transepidermal water loss rose from approximately 8.8 g m(-2) h(-1) before wIRA irradiation to 14.2 g m(-2) h(-1) after wIRA irradiation and skin hydration rose from 67 to 87 relative units. Skin surface temperature increased from 32.8 degrees C before wIRA to 36.4 degrees C after wIRA irradiation. DISCUSSION: The better penetration of the hydrophilic dye fluorescein after or during skin irradiation (modes B and C) can be explained by increased hydration of the stratum corneum by irradiation with wIRA. CONCLUSIONS: As most topically applied substances for the treatment of patients are mainly hydrophilic, wIRA can be used to improve the penetration of substances before or after application of substances - in the first case even of thermolabile substances - with a broad clinical relevance as a contact free alternative to an occlusive dressing. PMID- 19675736 TI - Antioxidants--friend or foe? PMID- 19675737 TI - Antioxidant vitamins and glucose-6-phosphate dehydrogenase deficiency in full term neonates. AB - OBJECTIVE: The mechanism by which glucose-6-phosphate dehydrogenase (G6PD) deficiency causes neonatal hyperbilirubinemia is not completely understood. However, the genetic disorder G6PD deficiency predisposes red blood cells to oxidative stress. The aim of this study was to establish the relationship between plasma antioxidant vitamin (E and C) levels and the development of hyperbilirubinemia in full-term neonates with deficient G6PD. METHODS: A total of 196 live birth neonates of healthy mothers were included in this study. Twelve of them were deficient in G6PD. In addition to demographic data, serum total bilirubin, hemoglobin, hematocrit, and vitamin E and C levels were measured on the first day after birth. RESULTS: Neonates with G6PD deficiency (n=7) who did not develop hyperbilirubinemia (mean serum bilirubin level of 70.8+/-23 micromol/l, median 71.8) and neonates with G6PD deficiency (n=4) who developed hyperbilirubinemia (mean serum bilirubin level of 226.7+/-79 micromol/l, median 233.4) on the first day of life had similar gestational weights and age. The second group, however, had lower hemoglobin and hematocrit as well as plasma vitamin C and E levels. None of these results showed significant difference. CONCLUSION: The results of the present study indicate that red blood cell hemolysis as a result of inadequate antioxidants system in G6PD-deficient neonates is not the only contributing factor for hyperbilirubinemia. PMID- 19675738 TI - Improvement of wound healing by water-filtered infrared-A (wIRA) in patients with chronic venous stasis ulcers of the lower legs including evaluation using infrared thermography. AB - BACKGROUND: Water-filtered infrared-A (wIRA) is a special form of heat radiation with a high tissue-penetration and with a low thermal burden to the surface of the skin. wIRA is able to improve essential and energetically meaningful factors of wound healing by thermal and non-thermal effects. AIM OF THE STUDY: prospective study (primarily planned randomised, controlled, blinded, de facto with one exception only one cohort possible) using wIRA in the treatment of patients with recalcitrant chronic venous stasis ulcers of the lower legs with thermographic follow-up. METHODS: 10 patients (5 males, 5 females, median age 62 years) with 11 recalcitrant chronic venous stasis ulcers of the lower legs were treated with water-filtered infrared-A and visible light irradiation (wIRA(+VIS), Hydrosun radiator type 501, 10 mm water cuvette, water-filtered spectrum 550-1400 nm) or visible light irradiation (VIS; only possible in one patient). The uncovered wounds of the patients were irradiated two to five times per week for 30 minutes at a standard distance of 25 cm (approximately 140 mW/cm(2) wIRA and approximately 45 mW/cm(2) VIS). Treatment continued for a period of up to 2 months (typically until closure or nearly closure of the ulcer). The main variable of interest was "percent change of ulcer size over time" including complete wound closure. Additional variables of interest were thermographic image analysis, patient's feeling of pain in the wound, amount of pain medication, assessment of the effect of the irradiation (by patient and by clinical investigator), assessment of feeling of the wound area (by patient), assessment of wound healing (by clinical investigator) and assessment of the cosmetic state (by patient and by clinical investigator). For these assessments visual analogue scales (VAS) were used. RESULTS: The study showed a complete or nearly complete healing of lower leg ulcers in 7 patients and a clear reduction of ulcer size in another 2 of 10 patients, a clear reduction of pain and pain medication consumption (e.g. from 15 to 0 pain tablets per day), and a normalization of the thermographic image (before the beginning of the therapy typically hyperthermic rim of the ulcer with relative hypothermic ulcer base, up to 4.5 degrees C temperature difference). In one patient the therapy of an ulcer of one leg was performed with the fully active radiator (wIRA(+VIS)), while the therapy of an ulcer of the other leg was made with a control group radiator (only VIS without wIRA), showing a clear difference in favour of the wIRA treatment. All mentioned VAS ratings improved remarkably during the period of irradiation treatment, representing an increased quality of life. Failures of complete or nearly complete wound healing were seen only in patients with arterial insufficiency, in smokers or in patients who did not have venous compression garment therapy. DISCUSSION AND CONCLUSIONS: wIRA can alleviate pain considerably (with an impressive decrease of the consumption of analgesics) and accelerate wound healing or improve a stagnating wound healing process and diminish an elevated wound exudation and inflammation both in acute and in chronic wounds (in this study shown in chronic venous stasis ulcers of the lower legs) and in problem wounds including infected wounds. In chronic recalcitrant wounds complete healing is achieved, which was not reached before. Other studies have shown that even without a disturbance of wound healing an acute wound healing process can be improved (e.g. reduced pain) by wIRA. wIRA is a contact-free, easily used and pleasantly felt procedure without consumption of material with a good penetration effect, which is similar to solar heat radiation on the surface of the earth in moderate climatic zones. Wound healing and infection defence (e.g. granulocyte function including antibacterial oxygen radical formation of the granulocytes) are critically dependent on a sufficient energy supply (and on sufficient oxygen). The good clinical effect of wIRA on wounds and also on problem wounds and wound infections can be explained by the improvement of both the energy supply and the oxygen supply (e.g. for the granulocyte function). wIRA causes as a thermal effect in the tissue an improvement in three decisive factors: tissue oxygen partial pressure, tissue temperature and tissue blood flow. Besides this non-thermal effects of infrared-A by direct stimulation of cells and cellular structures with reactions of the cells have also been described. It is concluded that wIRA can be used to improve wound healing, to reduce pain, exudation, and inflammation and to increase quality of life. PMID- 19675739 TI - Can posthysterectomy cystoscopy be utilized as a screening test for bladder cancer? AB - It is currently undisputed that cystoscopy is required after each hysterectomy to confirm the integrity of the urinary tract before leaving the operating room. Sometimes, secondary findings are detected on cystoscopy like interstitial cystitis and exophytic tumors. We are reporting a case of asymptomatic bladder cancer discovered incidentally during a posthysterectomy cystoscopy. The chance of a woman having this cancer is about 1 in 85. This demonstrates the additional merit of performing cystoscopy after hysterectomy as a screening test for bladder cancer. PMID- 19675740 TI - Management of fibromyalgia syndrome--an interdisciplinary evidence-based guideline. AB - The prevalence of fibromyalgia syndrome (FMS) of 1-2% in the general population associated with high disease-related costs and the conflicting data on treatment effectiveness had led to the development of evidence-based guidelines designed to provide patients and physicians guidance in selecting among the alternatives. Until now no evidence-based interdisciplinary (including patients) guideline for the management of FMS was available in Europe. Therefore a guideline for the management of fibromyalgia syndrome (FMS) was developed by 13 German medical and psychological associations and two patient self-help organisations. The task was coordinated by two German scientific umbrella organisations, the Association of the Scientific Medical Societies in Germany AWMF and the German Interdisciplinary Association of Pain Therapy DIVS. A systematic search of the literature including all controlled studies, systematic reviews and meta-analyses of pharmacological and non-pharmacological treatments of FMS was performed in the Cochrane Library (1993-12/2006), Medline (1980-12/2006), PsychInfo (1966-12/2006) and Scopus (1980 12/ 2006). Levels of evidence were assigned according to the classification system of the Oxford-Centre for Evidence Based Medicine. Grading of the strengths of recommendations was done according to the German program for disease management guidelines. Standardized procedures were used to reach a consensus on recommendations. The guideline was reviewed and finally approved by the boards of the societies involved and published online by the AWMF on april 25, 2008: http://www.uni-duesseldorf.de/AWMF/ll/041-004.htm. A short version of the guideline for patients is available as well: http://www.uni duesseldorf.de/AWMF/ll/041-004p.htm. The following procedures in the management of fms were strongly recommended: information on diagnosis and therapeutic options and patient-centered communication, aerobic exercise, cognitive and operant behavioural therapy, multicomponent treatment and amitriptyline. Based on expert opinion, a stepwise FMS-management was proposed. Step 1 comprises confirming the diagnosis and patient education and treatment of physical or mental comorbidities or aerobic exercise or cognitive behavioural therapy or amitriptyline. Step 2 includes multicomponent treatment. Step 3 comprises no further treatment or self-management (aerobic exercise, stress management) and/or booster multicomponent therapy and/or pharmacological therapy (duloxetine or fluoxetine or paroxetine or pregabalin or tramadol/aminoacetophen) and/or psychotherapy (hypnotherapy or written emotional disclosure) and/or physical therapy (balneotherapy or whole body heat therapy) and/or complementary therapies (homeopathy or vegetarian diet). The choice of treatment options should be based on informed decision-making and respect of the patients' preferences. PMID- 19675741 TI - GCP inspections in Germany and Europe following the implementation of the Directive 2001/20/EC. AB - BACKGROUND: The implementation of the Clinical Trials Directive 2001/20/EC and the Good Clinical Practice Directive 2005/28/EC fundamentally restructured and harmonized the conduct of clinical trials in Europe. GCP inspections - which affect study sites, laboratories, sponsors and contract research organizations (CRO) alike - make up an important part of these regulations. A common understanding of how these regulations apply in daily life is however not always ensured. METHODS: A working group of the Clinical Research/Quality Assurance subcommittee of the German Association of Research-Based Pharmaceutical Companies (VFA) was established to outline the regulatory requirements, the experience gathered with inspections by means of a survey and to set up guidance on how to manage an inspection. RESULTS AND CONCLUSIONS: The survey, conducted with the help of 15 pharmaceutical companies within the VFA, included a total of 224 inspections (74 inspections in Germany, 150 from other European countries). Most frequent findings in and outside Germany were related to "documentation" (40.5% vs. 21.3%), "investigational new drugs" (16.2% vs. 14.7%), "drug safety" (13.5% vs. 8%) and "application for a clinical trial authorization" (5.4% vs. 12%). From a German perspective, key findings of this working group were the necessity for a clear differentiation of responsibilities between national and federal as well as international authorities, a harmonization of inspection procedures and topics, and a clarification of whether pre-study/on-study and pre-approval/post-approval GCP inspections of the federal higher authority are included in the "Zentralstelle der Lander fur Gesundheitsschutz bei Arzneimitteln und Medizinprodukten" (ZLG) requirements. The survey illustrated, that inspections usually are conducted at the investigational site, and that most of the findings are well known and thus could be prevented by communicating and discussing audit results more intensely within study groups. Again, the survey illustrated, that a harmonization of inspections appears warranted. Finally a code of practice is provided that considers these findings and delivers a basis for a successful inspection whether at the sponsor or the GCP site. PMID- 19675742 TI - Undergraduate medical education in Germany. AB - The purpose of this article is to give international readers an overview of the organisation, structure and curriculum, together with important advances and problems, of undergraduate medical education in Germany. Interest in medical education in Germany has been relatively low but has gained momentum with the new "Regulation of the Licensing of Doctors" which came into effect in 2003. Medical education had required substantial reform, particularly with respect to improving the links between theoretical and clinical teaching and the extension of interdisciplinary and topic-related instruction. It takes six years and three months to complete the curriculum and training is divided into three sections: basic science (2 years), clinical science (3 years) and final clinical year. While the reorganisation of graduate medical education required by the new "Regulation of the Licensing of Doctors" has stimulated multiple excellent teaching projects, there is evidence that some of the stipulated changes have not been implemented. Indeed, whether the medical schools have complied with this regulation and its overall success remains to be assessed systematically. Mandatory external accreditation and periodic reaccreditation of medical faculties need to be established in Germany. PMID- 19675743 TI - Detection of HER-2/neu, c-myc amplification and p53 inactivation by FISH in Egyptian patients with breast cancer. AB - Breast cancer is a leading cause of cancer-related deaths in women worldwide. The clinical course of this disease is highly variable and clinicians continuously search for prognostic parameters that can accurately predict prognosis, and indicate a suitable adjuvant therapy for each patient. Amplification of the two oncogenes HER-2/neu and c-myc and inactivation of the tumor suppressor gene p53 are frequently encountered in breast carcinomas. The purpose of this study was to use the fluorescence in situ hybridization (FISH) for the assessment of HER-2/neu and c-myc amplification and p53 inactivation and to relate these molecular markers with the commonly used clinical and pathological factors. The study was conducted on 34 tissue samples obtained from 33 females and 1 male with breast carcinomas and 17 samples obtained from 16 females and 1 male with benign breast lesions. Results revealed that the level of HER-2/neu, c-myc and p53 in the malignant group was significantly increased as compared to the benign group. On relating the level of the molecular markers to clinicopathological factors, p53 was significantly associated with increased patient's age. The sensitivity of the investigated markers significantly increased with larger tumor size. Concerning tumor grade, HER-2/neu and p53 showed a significant increase in low-grade tumors whereas c-myc showed a highly significant increase in high-grade tumors. With regard to disease staging, HER-2/neu and c-myc were the only markers that showed significant increase at late stages of disease. p53 and HER-2/neu were significantly associated with positive lymph nodal status. A significant correlation was obtained between the levels of the three biomarkers to each other. Conclusively, the combination of HER-2/neu, c-myc and p53 can stratify patients into different risk groups. PMID- 19675744 TI - Phase I trial of metastatic renal cell carcinoma with oral capecitabine and thalidomide. AB - BACKGROUND: The highly vascular nature of renal carcinoma cells suggests that inhibition of angiogenesis may be beneficial in this disease. Thalidomide has been described as inhibitor of the fibroblast growth factor (FGF) and the vascular endothelial growth factor (VEGF). Therefore and in consideration of the promising response rates of the combination of IL-2, IFN-alpha and 5-FU [1] in metastatic renal cancer, we found it reasonable to test the combination of 5-FU and thalidomide. Thus, we conducted a phase I trial to determine safety, side effects and responses to such a treatment. METHODS: Patients with metastasized renal cell cancer after nephrectomy and progress after IL-2 and interferon treatment, received oral 5-FU at a dose of 1250 mg/qm(2) twice a day for two weeks, then after pausing a week, the oral application was restarted. In addition, oral thalidomide was applied constantly at a maximum dose of 400 mg/d. The combined therapy was given for three months. The primary endpoint was duration until disease progression, the secondary endpoint the response to treatment. Response was determined by CT scans three months after the end of treatment. RESULTS: In total, 12 male patients participated in the trial and received the combined oral therapy. Concerning clinical response, one mixed response (8%), a stable disease in 4/12 patients (33%) and progression was seen in 7 patients (58%). The survival from the start of the therapy showed a median of 21 months with three patients being alive. At present, the longest survival after the therapy is 51 months. CONCLUSIONS: The combination of oral 5-FU and thalidomide showed clinical response with tolerable side effects. Further studies will be required to assess the outcome of this treatment regimen. PMID- 19675745 TI - Carbon tetrachloride-induced liver disease in rats: the potential effect of supplement oils with vitamins E and C on the nutritional status. AB - The aim of the present investigation was to study the effects of olive oil (OO), corn oil (CO), and flaxseed oil (FO), with or without supplementation of vitamins E and C, on food intake, body weight gain %, liver weight to body weight %, total lipids, liver functions, and liver histology in male rats intoxicated with carbon tetrachloride (CCl(4)). Forty-two rats were divided into two main groups. The first main group was fed on basal diet (BD) as a negative control group (NC). The second main group received subcutaneous injections of CCl(4) in paraffin oil (50% v/v 2 ml/kg) twice a week to induce chronic damage in the liver. The group was then divided into six subgroups, three of which were fed on 4% unsupplemented oils (CO, FO, and OO) as positive control for the three oils used. The rest of the groups were fed on 4% of the same oils supplemented with vitamins E and C. The results of the flaxseed oil rat group indicate that supplementing vitamin E and C led to a significant reduction in the mean values of total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), and liver alanine amino transferase enzyme (ALT). Moreover, it caused an increase of the mean value of high-density lipoprotein cholesterol (HDL-C) as compared to the negative control group (NC). The olive oil group supplemented with the same vitamins showed a significant decrease in the mean value of serum TC and significant (P<0.05) increase in the mean value of serum HDL-C as compared to NC. The results of the corn oil group supplemented with vitamins showed a significant increase in the mean value of serum HDL-C as compared to the negative control group. The histology results confirmed that the group hepatically injured with CCl(4) treatment and fed on supplemented FO or OO showed apparently normal hepatocytes. CONCLUSION: The most effective treatment was observed with oils supplemented with vitamins E and C. Hierarchically FO achieved the best results compared to other additives, followed by OO and finally CO showing the least effective treatment among the observed groups. PMID- 19675746 TI - Rapid detection of methicillin-resistant Staphylococcus aureus directly from clinical samples: methods, effectiveness and cost considerations. AB - Methicillin-resistant Staphylococcus aureus (MRSA) isolates is a serious public health problem whose ever-increasing rate is commensurate with the pressure it is exerting on the healthcare system. At present, more than 20% of clinical S. aureus isolates in German hospitals are methicillin resistant. Strategies from low-prevalence countries show that this development is not necessarily inevitable. In the Scandinavian countries and the Netherlands, thanks to a rigorous prevention programme, MRSA prevalence has been kept at an acceptably low level (<1-3%). Central to these 'search and destroy' control strategies is an admission screening using several MRSA swabs taken from mucocutaneous colonisation sites of high-risk patients ('MRSA surveillance').It has also been reported that the speed with which MRSA carriage is detected has an important role to play, as it is a key component of any effective strategy to prevent the pathogen from spreading. Since MRSA culturing involves a 2-3 day delay before the final results are available, rapid detection techniques (commonly referred to as 'MRSA rapid tests') using PCR methods and, most recently, rapid culturing methods have been developed. The implementation of rapid tests reduces the time of detection of MRSA carriers from 48-72 to 2-5 h. Clinical evaluation data have shown that MRSA can thus be detected with very high sensitivity. Specificity however is sometimes impaired due to false-positive PCR signals occurring in mixed flora specimens. In order to rule out any false-positive PCR results, a culture screen must always be carried out simultaneously. The data provide preliminary evidence that a PCR assay can reduce nosocomial MRSA transmission in high-risk patients or high-risk areas, whereas an approach that screens all patients admitted to the hospital is probably not effective. Information concerning the cost-effectiveness of rapid MRSA tests is still sparse and thus the issue remains debated. PMID- 19675748 TI - HRPC: A need for multidisciplinary approach. PMID- 19675749 TI - Urology in ancient India. AB - The practice of medical and surgical measures in the management of urological ailments prevailed in ancient India from the Vedic era around 3000 BC. Subsequently in the Samhita period, the two stalwarts - Charaka in medicine and Susruta in surgery elevated the art of medicine in India to unprecedented heights. Their elaboration of the etiopathological hypothesis and the medical and surgical treatments of various urological disorders of unparalleled ingenuity still remain valid to some extent in our contemporary understanding. The new generation of accomplished Indian urologists should humbly venerate the legacy of the illustrious pioneers in urology of our motherland. PMID- 19675747 TI - Evaluation of clinical trials by Ethics Committees in Germany: experience of applicants with the review of requests for opinion of the Ethics Committees - results of a survey among members of the German Association of Research-Based Pharmaceutical Companies (VFA). AB - The review of requests for a positive opinion of the ethics committees (application procedure) as a requirement to start a clinical trial in Germany has been completely redesigned with the transposition of EU Directive 2001/20/EC in the 12(th) Amendment of the German Medicines Act in August 2004. The experience of applicants (sponsors, legal representatives of sponsors in the EU and persons or organizations authorized by the sponsors to make the application, respectively) in terms of interactions with the ethics committees in Germany has been positive overall, especially with respect to ethics committee adherence to the statutory timelines applicable for review of requests. However, inconsistencies between ethics committees exist in terms of the form and content of the requirements for application documents and their evaluation. With the objective of further improving both the quality of applications and the evaluation of those applications by ethics committees, a survey among members of the German Association of Research-Based Pharmaceutical Companies (VFA) was conducted from January to April 2008. Based on reasoned opinions issued by the respective ethics committee in charge of the coordinating principal investigator (coordinating ethics committee), the type and frequency of formal and content related objections to applications according to section sign 7 of the German Good Clinical Practice (GCP) Regulation were systematically documented, and qualitative and quantitative analyses performed. 21 out of 44 members of the VFA participated in the survey. 288 applications for Phase I-IV studies submitted between January and December 2007 to 40 ethics committees were evaluated. This survey shows that about one in six applications is incomplete and has formal and/or content objections, respectively, especially those that pertain to documents demonstrating the qualification of the investigator and/or suitability of the facilities. These objections are attributable to some extent to the differing and/or unclear requirements of the individual ethics committees on the content and comprehension of the submission documents. However, applicants also need to pay more attention to the completeness and validity of the submission documents. The majority of content-related objections apply to the patient information and consent documents and study protocols submitted. Applicants on average acted upon only 3 out of 4 objections, for various reasons: the relevant information was already given in the submitted documents, but had not been taken into consideration by the ethics committees; objections were not applicable; objections lacked a legal basis. In such cases the applicants made reference to the specific information already submitted or gave reasons for not acting on the objection. This course of action was accepted by the ethics committees, with few exceptions. The survey sheds light on the existing inconsistencies in the evaluations of applications by the various ethics committees and suggests ways in which the existing constructive dialogue between applicants and ethics committees may provide a basis to further harmonize both the requirements regarding form and content of application documents, and the criteria for evaluation of applications by ethics committees within the legal framework. PMID- 19675750 TI - Vasectomy: A simple snip? AB - Vasectomy is one of the most common forms of permanent sterilization methods currently in use and has a failure rate of <1% in most reported series. Since failure of vasectomy may result in pregnancy, adequate counseling is essential. Couples are advised that an analysis of a semen specimen after vasectomy is required to confirm success before the use of alternative contraception is abandoned. However, measuring the success of vasectomy is complicated by a lack of consistency with regards to both the number and timing of tests and the end points accepted. MATERIALS AND METHODS: A Medline search was used to identify manuscripts dealing with vasectomy, with specific attempts to identify protocols designed to confirm sterility. RESULTS AND CONCLUSION: Vasectomy is one of the most reliable permanent methods of contraception. However, despite its popularity, certain issues pertaining to the procedure remain unresolved. Debate continues over the relative merits of the various techniques of isolating and sealing the vasal ends. Postoperative complication rates remain minimal regardless of the technique used, and no single strategy attempting to maximize patient compliance with postoperative semen analysis has enjoyed unmitigated success. Long-term consequences, other than regret, are rare. PMID- 19675751 TI - Comparison of endopyelotomy and laparoscopic pyeloplasty for poorly functioning kidneys with ureteropelvic junction obstruction. AB - Endopyelotomy and laparoscopic pyeloplasty are established procedures for ureteropelvic junction obstruction (UPJO) and historically a high failure rate has been observed in poorly functioning units with UPJ obstruction. The aim of this study is to compare the results of laparoscopic pyeloplasty with endopyelotomy in poorly functioning renal units, i.e., GFR under 25 ml/min. MATERIALS AND METHODS: Retrospective analysis of all the patients who underwent either laparoscopic pyeloplasty or endopyelotomy for ureteropelvic junction obstruction in poorly functioning units between January 1998 and June 2005 was done. Follow-up renal scans, done at three, six, 12 months and yearly thereafter, were studied. Success was defined as symptomatic relief and/ or improvement in function (10% over baseline) in renal scan. RESULTS: There were 23 patients in the endopyelotomy group and 15 patients in the laparoscopic pyeloplasty group with mean age of 25.3 years (9-53) and 26 years (10-44), respectively. Mean pelvic volume was 41.2 8cc +/- 9.5 and 39.1cc +/- 9.85 in the endopyelotomy group and laparoscopic pyeloplasty group, respectively. Mean preoperative GFR was 17.4 +/- 5.7 ml/min and 21 +/- 4.5 ml/min in the endopyelotomy group and laparoscopic pyeloplasty group, respectively and mean postoperative GFR was 21 +/- 3.5 ml/min and 22 +/- 3.9 ml/min, respectively. Eighteen and 11 patients were symptomatic in ethe ndopyelotomy group and laparoscopic pyeloplasty group, respectively while symptomatic improvement was seen in 14 and 11 patients, respectively. Mean follow up was 12 months in the laparoscopy group and 28 months in the endopyelotomy group. Success rate was better for laparoscopic surgery group (15/15 = 100%) than for endopyelotomy (18/23 = 78.26%). CONCLUSIONS: Though the improvement in renal function is less in patients with UPJO with poorly functioning kidneys undergoing endopyelotomy or laparoscopic pyeloplasty, laparoscopic pyeloplasty gives better results in the form of symptomatic relief; however, renal function remains stable whichever the approach chosen. PMID- 19675752 TI - Which surgery is 'best' for patients with PUJ obstruction in a poorly functioning kidney? PMID- 19675753 TI - Endopyelotomy vs. laparoscopic pyeloplasty. PMID- 19675754 TI - Prediction of biochemical failure in localized carcinoma of prostate after radical prostatectomy by neuro-fuzzy. AB - OBJECTIVE: To predict biochemical failure in localized prostate cancer after radical prostatectomy using preoperative variables. MATERIALS AND METHODS: Twenty six patients of early carcinoma of prostate underwent open retropubic radical prostatectomy from June 2002 to June 2006. Preoperative variables included age, family history, digital rectal examination, serum prostatic specific antigen (S. PSA), prostate biopsy Gleason score, MRI of pelvis variables like periprostatic extension, seminal vesical invasion, weight of gland and pathological stage. With application of neuro-fuzzy, these variables were fed into system as input and output, that is S. PSA at six months (predicted value) was calculated. Neuro fuzzy system is a system to combine fuzzy system with learning techniques derived from neural networks. Here, we applied Takagi Sugeno Kang model (TSK) due to its close solution to our aim. All the patients were followed up for a minimum of six months. At six month S. PSA of all patients was done (observed value). Predicted and observed values were compared. RESULT: Predicted and observed values were plotted on 1:1 slop line. Coefficient of correlation was 0.9935. CONCLUSION: Coefficient of correlation is close to one. It indicates that the neuro-fuzzy is accurate in predicting biochemical failure in localized carcinoma of prostate after radical prostatectomy. PMID- 19675755 TI - Open nephron-sparing surgery in renal tumors with normal contralateral kidney: A single centre experience of 8 years. AB - INTRODUCTION: We present our eight-year experience with open nephron-sparing surgery (NSS) in renal tumors with contralateral normal kidney to assess its oncological efficacy and safety. MATERIALS AND METHODS: Thirty-six patients undergoing open NSS for small localized renal tumors with normal contralateral kidney from January 1998 to August 2006 were studied regarding demographic, clinical and pathological characteristics along with long-term follow-up. RESULTS: The mean age was 48.28 +/- 9.5 years. The mean tumor size was 3.72 cm (range 1.5-6). The following surgeries were performed: Wedge resection-13, partial polar nephrectomy-15, segmental resection-eight. The following techniques were used for vascular control: clamping and cooling-eight, warm ischemia-12, a novel technique of serial encirclage-16. The mean warm ischemia time was 23.2 +/- 3.2 min. The mean operating time was 190.07 +/- 11.3 min. The mean estimated blood loss was 331 +/- 17.4 ml. The majority of renal tumors were renal cell carcinoma (97.22%). There were no positive surgical margins. There were no major intraoperative and postoperative complications. The mean follow up was 52.1 months (range 4-80) with no case showing progression to renal insufficiency (defined as serum creatinine > 2 mg/dl). There was only one local recurrence. However, four distant metastases were reported. The five-year cancer-specific survival, recurrence-free survival and overall survival were 94.4%, 88.88% and 86.11% respectively. CONCLUSIONS: In patients with solitary, small localized, unilateral renal tumors with normal contralateral kidney, elective open NSS is feasible, safe and provides excellent long-term local control and oncological efficacy with functional benefits. PMID- 19675756 TI - Nephron sparing surgery: A single institution experience. AB - OBJECTIVE: To report our experience in managing various benign and malignant renal tumors with nephron-sparing surgery. MATERIALS AND METHODS: Records of patients who underwent nephron-sparing surgery (NSS) either through open or laparoscopic approach between May 1997 and June 2006 at our institution were reviewed. Patient and tumor-related characteristics, treatment modality and complications were noted. RESULTS: There were 26 patients (29 renal units), including three with bilateral lesions who underwent nephron-sparing surgery. Mean age at surgery was 47.0 years (range 16-67 years). Mean tumor size was 4.7 cm (range 2-7.5 cm). Mean warm ischemia time was 41 min and 32.5 min, operative time 158 min and 186 min and blood loss 200ml and 85 ml in open (n=24) and laparoscopic approach (n=2) respectively. Complications were seen in five (19.2%) patients of whom two had postoperative bleeding requiring nephrectomy in one and angioembolization in another. One patient with persistent urinary leak required intervention. Local wound infection in one patient and incisional hernia in another were surgically managed. Histopathological profile revealed 13 (44.8%) benign lesions which included angiomyolipoma (eight), simple cyst (two), cortical adenoma (one), metanephric adenoma (one) and myelolipoma (one). The remaining 16 (55.2%) malignant lesions included renal cell carcinoma (15) and metastatic adenocarcinoma (one). At a mean follow-up of 38.6 months (range 1-91) no patient had local recurrence or distant metastasis. Cancer-specific survival was 100% and overall survival was 92.3%. CONCLUSIONS: Nephron-sparing surgery is a safe and effective alternative to nephrectomy in both benign and malignant lesions of the kidney. PMID- 19675757 TI - Nephron sparing surgery: Is here to stay. PMID- 19675758 TI - Prolene (mesh) bulbourethral sling in male incontinence. AB - OBJECTIVE: We present our preliminary results of bulbar urethral sling (single bolster) in treatment of postprostatectomy urinary incontinence (PPUI). MATERIALS AND METHODS: From May 2003 to June 2005, six patients with postprostatectomy urinary incontinence (transurethral resection of prostate in five patients and after open prostatectomy in one patient) underwent prolene mesh bulbar urethral sling surgery. Preoperative evaluation included physical examination, neurological assessment, stress cystogram and urethrocystoscopy. Urodynamic evaluation was done in all patients for abdominal leak point pressure and ruling out bladder pathology. RESULTS: Urodynamic studies did not demonstrate bladder instability in any patient. Mean abdominal leak point pressure was 43cm of water (range 26-80 cm of water). Mean duration of hospital stay was 3.2 days. Follow-up ranged from 6-22 months. Four patients out of six patients were completely dry till their last follow-up. One patient developed mild stress incontinence after one year of the surgery and required use of one to two pads per day. Mean pad use after surgery was 0.6 pads per day in comparison to mean pad usage of 6.4 pads per day preoperatively. One patient was over-continent after the procedure and required clean intermittent catheterization till last follow-up (six months). Mean cost of the procedure was $ 350+15. CONCLUSION: Prolene bulbar urethral sling (single bolster) is an economically effective option in patients with postprostatectomy urinary incontinence. PMID- 19675759 TI - Bulbourethral sling in male incontinence. PMID- 19675760 TI - Hormone refractory prostate cancer: Current understanding and future perspectives. PMID- 19675762 TI - Update in palliative management of hormone refractory cancer of prostate. AB - Hormone refractory prostate cancer (HRPC) is an incurable disease and as in the pressure sensitive adhesive era the median survival of patients is increasing, these men increasingly develop symptomatic problems as a result of advanced local and or metastatic disease during their progression to death. Recently, it has been shown that it is possible to improve survival in this group of patients with use of chemotherapy which reinforces the need of better options in palliative care. We discus the various clinical problems (Part I) and treatment options of palliative care (Part II) and try to formulate an action plan in this review. PMID- 19675764 TI - Use of bisphosphonates in advanced prostate cancer: Current status. PMID- 19675761 TI - Pathogenesis of prostate cancer and hormone refractory prostate cancer. AB - Prostate cancer is the second most common malignancy in males and the leading cause of cancer death. Prostate cancer is initially androgen dependent and relies upon the androgen receptor (AR) to mediate the effects of androgens. The AR is also the target for therapy using antiandrogens and LHRH analogues. However, all cancers eventually become androgen independent, often referred to as hormone refractory prostate cancer. The processes involved in this transformation are yet to be fully understood but research in this area has discovered numerous potential mechanisms including AR amplification, over-expression or mutation and alterations in the AR signaling pathway. This review of the recent literature examines the current knowledge and developments in the understanding of the molecular biology of prostate cancer and hormone refractory prostate cancer, summarizing the well characterized pathways involved as well as introducing new concepts that may offer future solutions to this difficult problem. PMID- 19675763 TI - Use of bisphosphonates in prostate cancer: Current status. AB - Bisphosphonates are a relatively newer class of drugs which have been used for the prevention of skeletal related complications of age related osteoporosis or metastatic disease from carcinoma breast or multiple myeloma. Their role in the management of prostate cancer is still under evolution. We hereby review the ongoing and some published randomized trials to define the role of bisphosphonates in various stages of prostate cancer. PMID- 19675765 TI - Chemotherapy for hormone-resistant prostate cancer: Where are we today? AB - Significant progress has been achieved in chemotherapy for hormone-resistant prostate cancer (HRPC) in the last five years. Although the disease was long considered to be chemoresistant, docetaxel-based regimens in particular have been shown to both palliate symptoms and prolong survival in HRPC patients. Docetaxel is now considered the best available chemotherapy for prostate cancer progressing on first-line hormonal treatment. Other cytotoxics including mitoxantrone, anthracyclines, vinorelbin and vinblastine can alleviate symptoms and improve progression-free survival in HRPC without affecting overall survival. The survival benefit from chemotherapy seen in randomized studies has been small or nonexistent. Results of a recent trial suggest that the survival benefit may have been underestimated as a result of crossover from the less active to the active arm. PMID- 19675766 TI - New therapeutic targets in the treatment of prostate cancer. AB - Androgen deprivation therapy has become the mainstay of the treatment of advanced prostate cancer, being used in every clinical setting of the disease, from neoadjuvant to metastatic disease. Despite success in controlling the disease in the majority of men, hormonal manipulations will eventually fail. New agents are being developed for patients with hormone refractory disease. Important advances in molecular oncology have improved our understanding regarding the cellular mechanisms that regulate cell death in the prostate. It is hoped that these new insights will lead to development of more efficacious and easy to tolerate therapies for cancer prostate. This review focuses on the current literature on tumor vaccines, angiogenesis inhibitors, antisense oligonucleotides, differentiation agents, cancer-specific genes, endothelial receptor antagonists, anti-apoptotic agents, agents acting on signaling pathways and androgen and estrogen receptors. PMID- 19675767 TI - Treatment algorithm in hormone-resistant prostate cancer: Practical guidelines. AB - Treatment of hormone-resistant prostate cancer can be a challenging situation. The first important step in treating this condition is to assess if one has achieved the castrate level or not. If the castrate levels are not achieved, attempt should be made to achieve so. If the castrate level is achieved, then androgen withdrawals may be of help. Supportive care, care of the clinical problems forms an integral part of the treatment. Cancer-specific chemotherapy is certainly an option in progressive disease. PMID- 19675768 TI - Laparoscopic resection of a leiomyoma of the seminal vesicle. AB - We present a case of a leiomyoma of the seminal vesicle that occurred in a 52 year-old man who presented with symptoms of bladder outlet obstruction. Prostate specific antigen was within normal limit. Computed tomography scan and magnetic resonance imaging revealed a mass in the patient's right seminal vesicle. Laparoscopic excision of the seminal vesicle tumor was performed successfully. The patient was discharged from the hospital on the fourth postoperative day. PMID- 19675769 TI - Benign fibrous histiocytoma of the bladder. AB - Mesenchymal tumors of the bladder are rare with leiomyoma accounting for most of these. We present a rare case of a bladder benign fibrous histiocytoma in a 52 year-old male. He presented with history of straining during micturition since two years. The magnetic resonance imaging revealed a large intravesical mass. The mass was excised. It weighed 600g, with a maximum dimension of 13cm. Histology was that of a benign fibrous histiocytoma. To our knowledge, only two cases of this tumor have been reported in the bladder so far. The clinical and pathological features are discussed. PMID- 19675770 TI - Hernia uterine inguinale with transverse testicular ectopia and mixed germ cell tumor. AB - Persistent mullerian duct syndrome is a rare disorder characterized by the presence of uterus and fallopian tube in 46XY phenotypic males and is ascribed to defects in the synthesis or action of anti-mullerian hormone. We report a rare case of hernia uterine inguinale, transverse testicular ectopia associated with mixed germ cell tumor of the testis with metastasis. Transverse testicular ectopia should be suspected preoperatively in patients who have unilateral inguinal hernia associated with contralateral nonpalpable testis. In such cases ultrasonography should be done prior to repair of hernia to evaluate the possible presence of mullerian structures and testicular malignancy, for better management. PMID- 19675771 TI - Adrenal oncocytoma masquerading as a functional tumor. AB - Adrenal oncocytoma is a rare entity, with 20 cases reported in literature. A functional oncocytoma is extremely rare. We present a case of adrenal oncocytoma in a hypertensive male who had elevated catecholamine levels, which improved after adrenalectomy with decrease in daily antihypertensive requirement. PMID- 19675772 TI - Primary hyperoxaluria and systemic oxalosis. PMID- 19675773 TI - EAU guidelines for management of penile cancer. PMID- 19675774 TI - Is incidence of prostate cancer higher in patients with end stage renal disease? PMID- 19675776 TI - Prostate cancer treatment does not increase the risk of cardiovascular events. PMID- 19675775 TI - Neoadjuvant androgen suppression in patients treated with high-intensity focused ultrasound therapy. PMID- 19675777 TI - Increasing use of nephron sparing surgery for T1a renal tumors. PMID- 19675778 TI - Virtual versus real cystoscopy. PMID- 19675780 TI - Worshiping false Gods. PMID- 19675779 TI - Does treatment rate impact the efficacy of extracorporeal shock wave lithotripsy for kidney or ureteral stones? PMID- 19675781 TI - Application of evidence-based urology in improving quality of care. AB - Evidence-based medicine requires use of the best available evidence for optimal patient care. Increasingly scarce resources and escalating demands on time have led to emphasis on effective treatment. Opinion is slowly yielding to high quality existent evidence. It is important that urologists adapt to these changes for them to deliver optimum care to the patients. This article discusses the levels of evidence, the nature of desirable evidence, means of assessing quality of clinical trials and meta-analysis and finally the practice of evidence-based urology with special reference to bedside evidence-based urology. PMID- 19675783 TI - Erectile dysfunction among hypertensive men in a rapidly developing country. AB - BACKGROUND: Previous studies have supported the association between hypertension and erectile dysfunction (ED). In addition, a significant correlation between hypertension in men and ED has been well established. OBJECTIVES: The aim of this study was to investigate the prevalence of ED, its severity and other sexual function domains in hypertensive patients in Qatar. DESIGN AND SETTING: Cross sectional study conducted between January and December 2006 at the Hamad General Hospital. SUBJECTS: Qatari and nonQatari nationals 25 to 75 years of age were approached to participate in this study, 642 (75.5%) gave their consent. MATERIALS AND METHODS: Qatari and nonQatari men aged 25 to 75 years who gave consent were interviewed face-to-face. Sexual function was evaluated with the International index of erectile function. RESULTS: A total of 642 (75.5%) men responded, mean age of subjects was 57.1 +/- 11.9 years. The prevalence of ED in hypertensive patients was 58.3%. 21.2% of hypertensive men reported severe ED, 20.7% moderate ED and 16.4% mild ED. The comorbidities and risk factors were significantly more prominent in ED patients than in those with normal ED for hypercholesterolemia (P=0.009), diabetes (P=0.022) for hypertriglyceridemia (P=0.033) and myocardial ischemia (P=0.042). CONCLUSIONS: Our results have shown a greater prevalence of ED in the hypertensive men in the population of Qatar. The presence of another risk factor in addition to elevated blood pressure can increase the likelihood of ED. PMID- 19675784 TI - Clinical evaluation of a simple uroflowmeter for categorization of maximum urinary flow rate. AB - OBJECTIVE: To evaluate the accuracy and diagnostic usefulness of a disposable flowmeter consisting of a plastic funnel with a spout divided into three chambers. MATERIALS AND METHODS: Men with lower urinary tract symptoms (LUTS) voided sequentially into a standard flowmeter and the funnel device recording maximum flow rate (Q(max)) and voided volume (V(void)). The device was precalibrated such that filling of the bottom, middle and top chambers categorized maximum input flows as <10, 10-15 and > 15 ml s(-1) respectively. Subjects who agreed to use the funnel device at home obtained readings of flow category and V(void) twice daily for seven days. RESULTS: A single office reading in 46 men using the device showed good agreement with standard measurement of Q(max) for V(void) > 150 ml (Kappa = 0.68). All 14 men whose void reached the top chamber had standard Q(max) > 15 ml s(-1) (PPV = 100%, NPV = 72%) whilst eight of 12 men whose void remained in the bottom chamber had standard Q(max) < 10 ml s( 1) (PPV = 70%, NPV = 94%). During multiple home use by 14 men the device showed moderate repeatability (Kappa = 0.58) and correctly categorized Q(max) in comparison to standard measurement for 12 (87%) men. CONCLUSIONS: This study suggests that the device has sufficient accuracy and reliability for initial flow rate assessment in men with LUTS. The device can provide a single measurement or alternatively multiple home measurements to categorize men with Q(max) < 15 ml s( 1). PMID- 19675782 TI - Premature ejaculation. AB - Premature ejaculation (PE) is a common male sexual disorder. Recent normative data suggests that men with an intravaginal ejaculatory latency time (IELT) of less than 1 minute have "definite" PE, while men with IELTs between 1 and 1.5 minutes have "probable" PE. Although there is insufficient empirical evidence to identify the etiology of PE, there is limited correlational evidence to suggest that men with PE have high levels of sexual anxiety and inherited altered sensitivity of central 5-HT (5-hydroxytryptamine, serotonin) receptors. Pharmacological modulation of the ejaculatory threshold using off-label daily or on-demand selective serotonin re-uptake inhibitors is well tolerated and offers patients a high likelihood of achieving improved ejaculatory control within a few days of initiating treatment, consequential improvements in sexual desire and other sexual domains. Investigational drugs such as the ejaculo-selective serotonin transport inhibitor, dapoxetine represent a major development in sexual medicine. These drugs offer patients the convenience of on-demand dosing, significant improvements in IELT, ejaculatory control and sexual satisfaction with minimal adverse effects. PMID- 19675785 TI - Role of non-contrast spiral computerized tomography in acute ureteric colic. AB - AIM: To evaluate the sensitivity and specificity of noncontrast helical computerized tomography (CT) in ureteric colic with comparative evaluation of KUB and ultrasonography (USG). SETTING: Tertiary care university hospital. MATERIALS AND METHODS: One hundred patients aged between 20 and 75 years referred from the emergency department as acute ureteric colic were evaluated with KUB and USG followed by noncontrast helical CT. RESULTS: Noncontrast helical CT was 91% sensitive and 98% specific in detecting urolithiasis compared to a sensitivity of 20% and 30% for KUB and USG and specificity of 94% and 98% respectively. CONCLUSION: Noncontrast helical CT is a very sensitive and specific investigation for evaluation of acute flank pain due to urolithiasis, besides helping in the detection of nonrenal causes of pain. PMID- 19675786 TI - Age-specific prostate specific antigen and prostate specific antigen density values in a community-based Indian population. AB - INTRODUCTION: We sought to determine the distribution of serum prostate specific antigen (PSA) levels in a community-based population in the state of Gujarat in India. MATERIALS AND METHODS: Community screening of subjects residing in Kheda and Anand districts of Gujarat was done with digital rectal examination (DRE), uroflowmetry and transabdominal sonography and serum PSA. In three villages transrectal sonography was done in addition to transabdominal sonography. All patients with a PSA of more than 4 ng/ml and / or abnormal DRE were evaluated. Biopsy-proven prostatic malignancy was detected in 20 patients. Subjects who did not follow up and who had biopsy-proven malignancy were excluded from the analysis, 1899 subjects were analyzed. RESULTS: One thousand seven hundred and eighty-seven (89.17%) had a PSA of <4 ngml, 180 (8.9%) had a PSA between 4-10 ng/ml, while 37 (1.8 %) had a PSA more than 10ng/ml. There was a statistically significant correlation between age and prostate volume (correlation coefficient 0.33) and between prostate volume and PSA (correlation coefficient 0.50). The age specific PSA values derived as the 95(th) percentile value were as follows, 40-49 years (0-2.1), 50-59 years (0-3.4), 60-69 years (0-4.2) and more than 70 years (0 5.0). The 95(th) percentile value for PSAD was 0.19. CONCLUSION: Indians need to have separate PSA reference ranges. The age-specific PSA ranges for men in the present study population are on the lower side and the prostate specific antigen density on the higher side. PMID- 19675788 TI - Clinical evaluation of efficacy and safety of cyclosporine (Imusporin) in renal transplant patients with stable graft function maintained on neoral or bioral. AB - OBJECTIVE: Previous pharmacokinetic studies have demonstrated bioequivalence of Imusporin (microemulsion preparation of cyclosporine, Cipla) to the innovator product Neoral (Novartis, Switzerland). This study was done to evaluate the clinical efficacy and safety of Imusporin in patients who have already undergone renal transplant and have stable graft function maintained on cyclosporine preparation other than Imusporin. MATERIALS AND METHODS: Twenty-two renal allograft recipients (mean age of 31.77 years, range 18-53 years), with stable graft function, previously on Neoral or Bioral were switched over to Imusporin after recording their relevant baseline clinical and biochemical parameters. These were repeated on 1, 4, 7, 15, 30 and 90 days after the start of therapy. Change in dosage required to maintain C2 levels at each visit were analyzed by paired sample t-test. Safety of the drug was assessed by the type and severity of adverse events developed during the therapy. Cost analysis was done assuming an average maintenance immunosuppression dose of 150 mg/day of cyclosporine. RESULTS: Twenty-one patients completed the study. One patient was lost to follow up. Mean C2 value before switchover was 894 +/- 208 ng/ml, which was not significantly different from the mean values of C2 after switchover therapy (P>0.30). Change in dosage required to maintain C2 levels was not significantly different from the baseline dose of 2.34 mg/ kg body weight (P>0.1). No patient developed graft rejection after switchover therapy at a median follow-up of 16 months (14-18 months). Mean baseline SCr was similar to SCr at day 90 (1.38 vs. 1.37 mg/dl, P=0.930). No severe adverse events were reported. Mild side-effects included headache (4), somnolence (2), dry mouth (5) and generalized fatigue (6). Use of Imusporin (Cipla, India) results in an annual savings of Rs. 19892 over Neoral (Novartis, Switzerland) and Rs. 2263 over Bioral (Panacea Biotech, India). CONCLUSIONS: Imusporin is clinically as safe and efficacious as other cyclosporine preparations available while significantly reducing the cost of treatment. PMID- 19675787 TI - Comparison of fluid absorption during transurethral resection of prostate and Holmium-Yag laser enucleation of benign adenoma of prostate using breath ethanol concentration. AB - OBJECTIVE: We conducted a study to detect, quantify and compare irrigation fluid absorption in transurethral resection of the prostate (TURP) and Holmium laser enucleation of the prostate (HoLEP), using BEC. MATERIALS AND METHODS: The study included 50 patients of lower urinary tract symptoms, secondary to benign enlargement of prostate. The patients were nonrandomly allocated to undergo TURP and HoLEP. Twenty-six patients underwent TURP and the remaining 24 underwent HoLEP. Sterile water tagged with 1% ethanol w/v was used for irrigation. Absorption was detected and quantified every 10min by BEC levels. Data was analyzed using standard nomograms. RESULTS: In HoLEP, 14/24 had no fluid absorption. The remaining 10/24 showed fluid absorption ranging from 95 ml to 300 ml. In TURP, all had fluid absorption ranging from 250-980 ml. Three TURP patients developed overt symptoms, while none did in the HoLEP group. CONCLUSIONS: Fluid absorption observed in our study in the HoLEP group was lower than in the TURP group. PMID- 19675789 TI - Recent updates in the management of urinary incontinence. PMID- 19675790 TI - Pelvic organ prolapse and stress urinary incontinence: A review of etiological factors. AB - Although they may present with significant morbidity, pelvic organ prolapse and stress urinary incontinence are mainly afflictions that affect quality of life. To appropriately treat these entities, comprehension of the various theories of the pathophysiology is paramount. Utilizing a Medline search, this article reviews recent data concerning intrinsic (i.e., genetics, postmenopausal status) and extrinsic factors (i.e., previous hysterectomy, childbirth) leading to organ prolapse or stress incontinence. PMID- 19675791 TI - Use of urodynamics prior to surgery for urinary incontinence: How helpful is preoperative testing? AB - It has not yet been definitively demonstrated that preoperative evaluation of women with stress urinary incontinence with urodynamic testing enhances presurgical counseling, more effectively models patients' expectations or improves postoperative outcome. Nonetheless, urodynamic testing is frequently utilized in the assessment of women with stress urinary incontinence and clearly accomplishes a number of goals when utilized for this purpose. For example, there are data to suggest that the risk of voiding dysfunction can be mitigated by utilizing data obtained from urodynamic testing to identify women more likely to void ineffectively after conventional stress incontinence procedures. Furthermore, it has been suggested though not proven, that patients with more severe forms of stress incontinence as identified by urodynamic testing, might be less likely to improve after surgery compared to others with more modest degrees of incontinence. Since urodynamic testing is invasive, costly and not always available, it is imperative that the usefulness of such testing be carefully explored and its utility appropriately defined. In this review, we discuss urodynamic techniques to assess stress urinary incontinence, particularly focusing on the ability of leak point pressure testing and urethral pressure profilometry to predict which patients would most likely benefit from surgery and which might be more likely to experience adverse events following surgery. PMID- 19675792 TI - Surgery for stress urinary incontinence in women: A 2006 review. AB - The surgical treatment of female stress urinary incontinence is a rapidly changing field. This review discusses recent advances in various injectables, minimally invasive techniques and open procedures. It particularly evaluates data from long-term outcome studies and describes peri- and postoperative complications from several procedures, such as bulking agents, tension-free vaginal tape and its modifications (TOT, TVT-O) as well as open and laparoscopic colposuspension. PMID- 19675793 TI - Complications of grafts used in female pelvic floor reconstruction: Mesh erosion and extrusion. AB - INTRODUCTION: Various grafts have been used in the treatment of urinary incontinence and pelvic prolapse. Autologous materials such as muscle and fascia were first utilized to provide additional anatomic support to the periurethral and pelvic tissues; however, attempts to minimize the invasiveness of the procedures have led to the use of synthetic materials. Complications such as infection and erosion or extrusion associated with these materials may be troublesome to manage. We review the literature and describe a brief overview of grafts used in pelvic floor reconstruction and focus on the management complications specifically related to synthetic materials. MATERIALS AND METHODS: We performed a comprehensive review of the literature on grafts used in pelvic floor surgery using MEDLINE and resources cited in those peer-reviewed manuscripts. The results are presented. RESULTS: Biologic materials provide adequate cure rates but have associated downfalls including potential complications from harvesting, variable tissue quality and cost. The use of synthetic materials as an alternative graft in pelvic floor repairs has become a popular option. Of all synthetic materials, the type I macroporous polypropylene meshes have demonstrated superiority in terms of efficacy and fewer complication rates due to their structure and composition. Erosion and extrusion of mesh are common and troublesome complications that may be managed conservatively with observation with or without local hormone therapy, with transvaginal debridement or with surgical exploration and total mesh excision, dependent upon the location of the mesh and the mesh type utilized. CONCLUSIONS: The ideal graft would provide structural integrity and durability with minimal adverse reaction by the host tissue. Biologic materials in general tend to have fewer associated complications, however, the risks of harvesting, variable integrity of allografts, availability and high cost has led to the development and use of synthetic grafts. Synthetic grafts have a tendency to cause higher rates of erosion and extrusion; however, these complications can be managed successfully. PMID- 19675794 TI - Conservative management of voiding dysfunction. AB - This review article discusses the efficacy of various conservative therapies in the management of voiding dysfunction with special reference to urinary incontinence. The article emphasizes the fact that conservative therapies have limited side effects and they do not jeopardize future treatment options. Behaviour therapy, pelvic floor therapy and biofeedback; electrical and magnetic stimulation are discussed here individually. Though there is unanimous agreement that these therapies improve quality of life, complete cure is rare. All therapies work better in conjunction with each other rather than in isolation. The review also highlights the need for randomized controlled trials of better methodology. PMID- 19675796 TI - Male incontinence: Pathophysiology and management. AB - Post-prostatectomy incontinence in men is a devastating condition. It impacts the quality of life profoundly. Various types of male sling procedures have been introduced over the years. Bone anchored male sling appears to be effective and safe in intermediate term follow up. It certainly more effective than collagen implant and may provide alternative treatment option in patient with mild to moderate incontinence. In short term, other novel procedures seem to be promising. In spite of new technology, artificial urinary sphincter continues to provide high patient satisfaction and cure rates. PMID- 19675795 TI - Refractory overactive bladder: Beyond oral anticholinergic therapy. AB - OBJECTIVES: In this review, we discuss the treatment of refractory overactive bladder (OAB) that has not adequately responded to medication therapy and we propose an appropriate care pathway to the treatment of OAB. We also attempt to address the cost of OAB treatments. MATERIALS AND METHODS: A selective expert review of the current literature on the subject of refractory OAB using MEDLINE was performed and the data is summarized. We also review our experience in treating refractory OAB. The role and outcomes of various treatment options for refractory OAB are discussed and combined therapy with oral anticholinergics is explored. Emerging remedies including intravesical botulinum toxin injection and pudendal neuromodulation are also reviewed, along with conventional surgical options. RESULTS: In general behavioral therapy, pelvic floor electrical stimulation, magnetic therapy and posterior tibial nerve stimulation (PTNS), have shown symptom decreases in 50-80% of patients with OAB. Depending on the study, combination therapy with oral anticholinergics seems to improve efficacy of behavioral therapy and PTNS in approximately 10-30%. In multicenter, long-term randomized controlled trials, sacral neuromodulation has been shown to improve symptoms of OAB and OAB incontinence in up to 80% of the patients treated. Studies involving emerging therapies such as pudendal serve stimulation suggest that there may be a 15-20% increase in efficacy over sacral neuromodulation, but long-term studies are not yet available. Another emerging therapy, botulinum toxin, is also showing similar success in reducing OAB symptoms in 80-90% of patients. Surgical approaches, such as bladder augmentation, are a last resort in the treatment of OAB and are rarely used at this point unless upper tract damage is a concern and all other treatment options have been exhausted. CONCLUSION: The vast majority of OAB patients can be managed successfully by behavioral options with or without anticholinergic medications. When those fail, neuromodulation or intravesical botulinum toxin therapies are successful alternatives for most of the remaining group. We encourage practitioners responsible for the care of OAB patients to gain experience with these options. More research is needed to assess the cost-effectiveness of various OAB treatments. PMID- 19675797 TI - Male incontinence: Pathophysiology and management. PMID- 19675798 TI - Meningomylocele: An update. AB - Therapy-resistant overactivity of detrusor or small capacity and poor compliance, will usually need to be treated by bladder augmentation. Ileal or colonic patches are used frequently for augmenting the bladder, with either intestinal segment appearing to be equally useful. Stomach is rarely used because of the associated complications, but it is the only available intestinal segment for patients with impaired renal function. Concerns regarding long-term effects of associated metabolic acidosis, including abnormalities in linear growth and bone metabolism are misplaced. Ureterocystoplasty offers an attractive urothelium-preserving alternative, avoiding the metabolic complications, mucus production and cancer risk of heterotopic epithelium associated with enterocystoplasty. Though ideal for patients with dilated ureter and nonfunctioning kidney, in patients with functioning kidney it carries added risks associated with transuretero ureterostomy, mainly obstruction. Ureteral dilatation in meningomyelocele patients is avoidable with proper follow-up and treatment. Therefore they rarely should be candidates for this operation. Alternative urothelium-preserving techniques, such as auto augmentation and seromuscular cystoplasty, have not proven to be as successful as standard augmentation with intestinal segment. Work is in progress on various bioengineering techniques to culture and combine bladder cells in tissue culture for regeneration. Early efforts are exciting, but preliminary. PMID- 19675800 TI - What is the prevalence of overactive bladder symptoms in a lower socioeconomic female population?: A suggestion for a study in India. AB - Although it is suggested that in the United States overactive bladder affects one out of six individuals, this estimation may represent a subset of the population. Using a Pubmed literature search, many studies do not address those in a lower socioeconomic strata and the prevalence of overactive bladder may be higher. Overactive bladder symptoms may be under-reported in this population due to social stigma, lack of education or inaccessibility to medical care. This paper proposes to perform an epidemiological study incorporating validated incontinence questionnaires to assess the prevalence of overactive bladder symptoms among Indian women. PMID- 19675801 TI - Paecilomyces pyelonephritis in a patient with urolithiasis. AB - Fungal organisms are increasingly implicated in nosocomial urinary tract infections. Although Candida, Mucor and Aspergillus are the most commonly identified species, rare fungi are also occasionally observed to infect humans. Misidentification of the organism could result in treatment with an inappropriate antifungal agent, which could result in a florid fungal pyelonephritis. We report the occurrence of fungal pyelonephritis in a patient with stone disease secondary to Paecilomyces variotii. This case report emphasizes the need for an accurate identification of the organism and early and appropriate treatment. PMID- 19675802 TI - An unusual case of urinary tract obstruction due to imperforate hymen in an 11 month-old infant. AB - Variations in the anatomy of the hymen are common and imperforate hymen is an extreme manifestation. In spite of the recommendations for early inspection of the external genitalia, hymenal malformations escape diagnosis until the time of menarche. Rarely, female infants with imperforate hymen present with urologic complications. We would like to present an unusual case of urinary retention in an 11-month-old female infant due to imperforate hymen. This infant was successfully treated surgically. PMID- 19675799 TI - Vesicovaginal fistula: An update. AB - Vesicovaginal fistula (VVF) has been a social and a surgical problem for centuries and remains a challenge to surgeons. Though advances have been made in the understanding of the etiology, diagnostic procedures and management of these fistulae, controversies still exist over the ideal approach and time to repair. This review was undertaken to look into the recent literature with regard to the timing and surgical approach to VVF repair. The literature search was done using the Medline database with keywords: vesicovaginal fistula, laparoscopy and robotic repair filtered for the last 5 years. PMID- 19675803 TI - Failure of sonography to visualize a kidney affected by emphysematous pyelonephritis. AB - We describe a diabetic patient who presented with acute renal failure as a result of acute bilateral emphysematous pyelonephritis. Initially, both an abdominal X ray examination and a renal sonogram were unremarkable. Two days later, however, the previously visualized right kidney could not be demonstrated again by a repeat renal sonogram. A computed tomogram and a repeat abdominal X-ray study confirmed the diagnosis of emphysematous pyelonephritis. To our knowledge this is one of the few described cases of emphysematous pyelonephritis distinguished by a sonogram's inability to visualize a kidney because of interference with imaging by the gas produced by the bacteria that are responsible for the pyelonephritic infection. PMID- 19675804 TI - An unusual complication of silver nitrate therapy for chyluria. AB - Chyluria is a common disorder in the tropics. In our country it is most commonly managed by instillation of silver nitrate. We report a patient who developed severe perinephric hematoma due to pseudoaneurysm of renal artery following silver nitrate instillation. He was managed by angioembolisation followed by drainage of infected perinephric hematoma. We discuss various modalities of treatment of chyluria including complications of silver nitrate therapy. PMID- 19675805 TI - An unusual complication of silver nitrate therapy for chyluria. PMID- 19675806 TI - Posterior urethral polyps and review of literature. AB - Urethral polyp is a rare finding in young children. Fibroepithelial polyps of the urethra are usually diagnosed during the first decade of life. They present with obstruction, voiding dysfunction and hematuria. They can be associated with other congenital urinary tract anomalies. They are usually benign fibroepithelial lesions with no tendency to recur and are treated by surgical ablation, fulguration or laser therapy. PMID- 19675807 TI - Penile gangrene in diabetes mellitus with renal failure: A poor prognostic sign of systemic vascular calciphylaxis. AB - Penile gangrene associated with chronic renal failure is very uncommon. A 52-year old man with diabetes mellitus, diffuse atherosclerosis, ischemic cardiomyopathy and end-stage renal disease presented with blackening of distal penis for 10 days. His general condition was poor and gangrene of prepuce and glans was noted. Doppler and magnetic-resonance angiography revealed bilateral internal iliac artery obstruction. He underwent trocar suprapubic cystostomy and was planned for partial penectomy. But he died of severe diabetic complications in the interim period. Penile gangrene is a manifestation of widespread vascular calcifications associated with end-stage renal disease and is a marker of poor prognosis. PMID- 19675808 TI - Ureteral frozen section analysis during radical cystectomy: Do margins matter? PMID- 19675810 TI - Search for new alpha1a-adrenoceptor-selective antagonist for treating lower urinary tract symptoms associated with benign prostatic hyperplasia. PMID- 19675809 TI - Preservation of urethra devoid of corpus spongiosum in patients undergoing urethroplasty. PMID- 19675811 TI - Which one is better, Wallace or Bricker? PMID- 19675812 TI - Are we along the path for laparoscopic partial nephrectomy? PMID- 19675814 TI - Transurethral ethanol ablation for symptomatic benign prostatic hyperplasia. PMID- 19675813 TI - T1 bladder cancer on restaging transurethral resection should be treated with immediate cystectomy. PMID- 19675815 TI - Role of simultaneous fine needle aspiration cytology of palpable inguinal lymph node and primary tumor biopsy in carcinoma of the penis. PMID- 19675817 TI - Prophylactic application of levosimendan in cardiac surgical patients with severe left ventricle dysfunction. AB - BACKGROUND: Levosimendan improves myocardial contractility, and increases systemic, pulmonary and coronary vasodilation. The present study investigates the perioperative hemodynamic effects of a prophylactic infusion of levosimendan in high-risk cardiac surgery patients with severe left ventricle dysfunction, and compares short-term clinical outcomes with a historical control group in which dobutamine and milrinone were used. PATIENTS AND METHODS: A retrospective, observational study was performed in 10 adult patients with EuroSCOREs greater than six and severe left ventricle dysfunction. In the study group, prophylactic levosimendan infusion was started after the induction of general anesthesia (bolus 24 mug/kg over 10 min; continuous infusion 0.1 mug/kg/min over the next 24 h). The historical control group (12 patients) was treated with dobutamine and milrinone. The hemodynamic measurements were performed at eight predetermined time points. Short-term clinical outcomes recorded in both groups were the length of intensive care unit stay, the need for dialysis, the length of hospital stay, predicted mortality (logistic EuroSCORE) and 30-day mortality. RESULTS: Hemodynamic measurements performed in the study group revealed an increase in cardiac index 30 min after levosimendan infusion was started. Cardiac performance was sufficient throughout the entire perioperative period. The length of intensive care unit stay was not different between the groups, but the length of hospital stay (10.4+/-5.0 versus 26.6+/-17.6 days; P=0.01) and 30-day mortality (0% versus 41.7%; P=0.04) were significantly lower in levosimendan-treated patients. CONCLUSIONS: Levosimendan seemed to be an effective choice for preventing left ventricle failure in high-risk cardiac surgical patients with severe left ventricle dysfunction compared with patients receiving dobutamine and milrinone. PMID- 19675816 TI - The use of risk scores for stratification of non-ST elevation acute coronary syndrome patients. AB - OBJECTIVE: To review the methods available for the risk stratification of non-ST elevation (NSTE) acute coronary syndrome (ACS) patients and to evaluate the use of risk scores for their initial risk assessment. DATA SOURCES: The data of the present review were identified by searching PUBMED and other databases (1996 to 2008) using the key terms "risk stratification", "risk scores", "NSTEMI", "UA" and "acute coronary syndrome". STUDY SELECTION: Mainly original articles, guidelines and critical reviews written by major pioneer researchers in this field were selected. RESULT: After evaluation of several risk predictors and risk scores, it was found that estimating risk based on clinical characteristics is challenging and imprecise. Risk predictors, whether used alone or in simple binary combination, lacked sufficient precision because they have high specificity but low sensitivity. Risk scores are more accurate at stratifying NSTE ACS patients into low-, intermediate- or high-risk groups. The Global Registry of Acute Cardiac Events risk score was found to have superior predictive accuracy compared with other risk scores in ACS population. Treatments based according to specific clinical and risk grouping show that certain benefits may be predominantly or exclusively restricted to higher risk patients. CONCLUSION: Based on the trials in the literature, the Global Registry of Acute Cardiac Events risk score is more advantageous and easier to use than other risk scores. It can categorize a patient's risk of death and/or ischemic events, which can help tailor therapy to match the intensity of the patient's NSTE ACS. PMID- 19675818 TI - Takotsubo cardiomyopathy complicated by high-grade atrioventricular block: A report of two cases. AB - The present report describes two female patients aged 39 and 57 years who experienced loss of consciousness and chest pain due to high-grade atrioventricular block. Both patients demonstrated noncontraction centred on the cardiac apex and excessive contraction at the cardiac base on cardiac ultrasonography and left ventriculography, but neither of them demonstrated any significant stenotic lesions on coronary angiography. Furthermore, neither patient showed elevated serum biomarkers of cardiac injury or serum viral antibodies. In a repeat left ventriculogram two weeks later, the left ventricular wall motion disorder had improved in both patients. Based on these findings, the patients were diagnosed with takotsubo cardiomyopathy. Because the high-grade atrioventricular conduction disorder did not improve in spite of the improvement of left ventricular wall motion disorder, permanent pacemaker implantation was performed. It is extremely rare for takotsubo cardiomyopathy to be complicated by high-grade atrioventricular block. In the present study, both patients had takotsubo cardiomyopathy complicated by high-grade atrioventricular block and eventually underwent permanent pacemaker implantation. PMID- 19675819 TI - Successful use of a guide wire to engage an occluded vessel following failure of conventional catheterization techniques in a patient with acute myocardial infarction. AB - The present report describes the case of a 43-year-old woman with acute inferior myocardial infarction, in whom the culprit vessel was posteromedially located, and various diagnostic or guide catheters could not be engaged. The culprit vessel was directly cannulated using a guide wire, which allowed for a successful coronary intervention. PMID- 19675821 TI - The effects of renovascular hypertension on repolarization of ventricular epicardium. AB - BACKGROUND AND OBJECTIVE: Alterations in the recovery sequence of hypertrophied myocardium favour the development of cardiac arrhythmias. The aim of the present study was to investigate apex-to-base and interventricular heterogeneities in the duration of epicardial ventricular repolarization in rats with renovascular hypertension. METHOD: Renovascular hypertension was induced in six Wistar rats by constricting the left renal artery for one month. Six sham-operated Wistar rats served as normotensive controls. Epicardial mapping was performed using 32 unipolar leads distributed over the apex and base of the heart ventricles under sinus rhythm. Activation-recovery intervals (ARIs) were calculated from electrograms. RESULTS: The ratio of left ventricular weight to body weight was increased in hypertensive rats compared with controls. In control rats, ARIs at the base of both ventricles were shorter than those at the apex. In hypertrophied hearts, ARIs were prolonged on both the left and right ventricular epicardium. Heterogeneous prolongation was observed via reduced apex-to-base differences in ARIs and increased interventricular differences, with a trend toward increasing dispersion of ARIs. In rats with renovascular hypertension, nonuniform prolongation of epicardial ARIs on both ventricles and the changes in the ARI distribution resulted in a reduction of the repolarization time gradient between the ventricles. CONCLUSION: Nonuniformly prolonged ARIs across the ventricular epicardium and the interventricular electrical inhomogeneity in rats with renovascular hypertension should be considered when interpreting the T wave alterations together with the reduction of the transmural and apex-to-base repolarization gradients. PMID- 19675822 TI - Effect of a low dose of proteasome inhibitor on cell death and gene expression in neonatal rat cardiomyocyte cultures exposed to anoxia-reoxygenation. AB - BACKGROUND: Recent data suggest that low concentrations of proteasome inhibitors (PIs) are cytoprotective in models of ischemia-reperfusion injury, but the underlying mechanisms of this effect still remain unclear. AIM: To investigate the effect of 100 nM of clasto-lactacystin beta-lactone on cell death and gene expression in neonatal rat cardiomyocytes exposed to anoxia-reoxygenation. METHODS: Fluorescent microscopy and real-time polymerase chain reaction were used to detect different types of cell death and gene expression, respectively, in neonatal rat cardiomyocyte cultures exposed to anoxia-reoxygenation. RESULTS: It was shown that a low dose of clasto-lactacystin beta-lactone protected the cells against anoxia-reoxygenation injury by a reduction in the number of necrotic and apoptotic cells. The number of autophagic cells was greatly increased by proteasomal inhibition. The PI increased the heat shock protein 70 messenger RNA expression twofold and slightly reduced the expression of heat shock protein 90 gene. The expression of the FK506 binding protein 12-rapamycin associated protein gene was increased 1.57-fold on PI application. The B-cell lymphoma 2 gene expression was unaffected by the use of clasto-lactacystin beta-lactone in low dose. CONCLUSION: Although PIs are injurious, they may be cardioprotective in low doses; ie, they do not result in cell death. Moreover, PIs initiate the protective mechanisms that prevent cell damage by changing the expression of several genes. PMID- 19675820 TI - Insulin resistance and endothelial dysfunction: Are epoxyeicosatrienoic acids the link? AB - Epoxyeicosatrienoic acids (EETs), the cytochrome P450 epoxygenase metabolites of arachidonic acid, are potent vasodilators and are believed to be the endothelium derived hyperpolarizing factor in a number of vascular beds. In addition, EETs may play a role in the secretion and action of insulin and the metabolism of carbohydrates and lipids. Pharmacological manipulation of EETs may be a useful therapeutic approach for disease states such as hypertension, diabetes mellitus and the metabolic syndrome. EET mimetics and antagonists and drugs that increase EET synthesis or decrease their degradation are currently under investigation. The cellular mechanism of action of EETs appears to be complex and is being intensively studied by a number of investigators. In the present article, EET production, metabolism, isomerism and vasodilatory effects will be reviewed and potential mechanisms of action discussed. The role of EETs in insulin secretion and sensitivity and their implication in diabetes mellitus and the metabolic syndrome will also be reviewed. Drugs affecting EET bioavailability and action may be promising agents to use to treat hypertension/insulin resistance. The effects of these agents in experimental vascular disorders will also be discussed. PMID- 19675823 TI - Heart sounds are altered by open cardiac surgery. AB - BACKGROUND AND OBJECTIVE: Patients have reported that they perceive their own heart sounds differently after open cardiac surgery than before the surgery. The present study was designed to investigate whether changes in heart sounds can be quantitatively measured. METHOD: Heart sounds were recorded from 57 patients undergoing coronary artery bypass graft (CABG) surgery and from a control group of 10 subjects. The so-called Hjorth descriptors and the main frequency peak were compared before and after surgery to determine whether the characteristics of the heart sounds had changed. RESULTS: At a group level, the first heart sound was found to be significantly different after CABG surgery. Generally, the heart sounds shifted toward a lower frequency after surgery in the CABG group. No significant changes were found in the control group. CONCLUSIONS: Heart sounds are altered after CABG surgery. The changes are objectively quantifiable and may also be subjectively perceived by the patients. PMID- 19675824 TI - Perspectives on interactions between paediatricians and the pharmaceutical industry. PMID- 19675825 TI - Exclusive breastfeeding should continue to six months. PMID- 19675826 TI - Transfusion and risk of infection in Canada: Update 2005. PMID- 19675827 TI - Complementary and alternative medicine in Canadian children: A call for action. AB - Complementary and alternative medicine (CAM) use, including paediatric use, is common. The Hospital for Sick Children Foundation organized a priority-setting forum regarding paediatrics and the use of CAM/natural health products in November 2001. Four priority areas were identified: the creation of a national paediatric research network and/or centre of excellence in CAM; support for experiential learning, education, and training; completion of a needs/use assessment; and facilitation of knowledge transfer. These priority areas are discussed in the context of research, education, knowledge transfer and health systems. The present article represents a call for action for Canadian funding agencies to support the development of an evidence base in paediatric CAM. PMID- 19675828 TI - Assessing the evidence for the use of chiropractic manipulation in paediatric health conditions: A systematic review. AB - OBJECTIVE: To review the biomedical literature up to and including 2003, and determine the extent of the evidence related to the therapeutic application of chiropractic manipulation for paediatric health conditions. No critical appraisal of the evidence is undertaken. DATA SOURCES: The indexed manual therapy sector including medical, chiropractic, physiotherapy, naturopathic and osteopathic literature was searched. This included PubMed; the Manual, Alternative, and Natural Therapy Index System; the Cumulative Index to Nursing and Allied Health Literature; the Index to Chiropractic Literature; the Paediatric Economic Database Evaluation Project; the Cochrane Library; the Canadian Coordinating Office for Health Technology Assessment database; and the Agency for Healthcare Research and Quality database. Other resources included research conference and symposium proceedings, and the references of identified studies. RESULT: The search identified 1731 articles, of which 166 met the eligibility criteria. Two reviewers determined by consensus each citation's appropriate level on the strength of evidence scale. There was one systematic review, nine randomized controlled trials, one observational study, 141 descriptive case studies and 14 conference abstracts. SUMMARY: Health claims made by practitioners regarding the application of chiropractic manipulation as a health care intervention for paediatric health conditions are, for the most part, supported by low levels of scientific evidence. Chiropractors, in particular, employ manipulation for the treatment of a wide variety of paediatric health conditions. The evidence rests primarily with clinical experience, descriptive case studies and a few randomized controlled trials. There is a need for more rigorous scientific inquiry to examine the value of manipulative therapy in the treatment of paediatric conditions. To advance the health interests of paediatric patients, health care decisions made on the basis of expert opinion or clinical experience must integrate the best research evidence available from high-quality, scientific studies. PMID- 19675830 TI - Does Lactobacillus GG prevent antibiotic-associated diarrhea in children?: Part A: Evidence-based answer and summary. PMID- 19675829 TI - Three patients and their drugs: A parallel case paper on paediatric opiate use and withdrawal. AB - Clinicians are increasingly aware of the important role that opioids play in the management of a variety of paediatric conditions. Frequently, clinicians encounter challenges in initiating opioid therapy and, then, in weaning paediatric patients off opioids. In the present article, three different cases (an infant and two adolescents) are used to illustrate why and how opiates may be used in paediatrics and how they can be discontinued. The presentations include neonatal abstinence syndrome, chronic headache and cystic fibrosis. The cases are meant to emphasize the distinctions rather than the similarities among the cases, which is why this is called a case parallel, rather than a case series. The article downplays the use of rigid rules in managing patients on opioids, and emphasizes a flexible and patient-centred approach. PMID- 19675831 TI - Does Lactobacillus GG prevent antibiotic-associated diarrhea in children?: Part B: Clinical commentary. PMID- 19675832 TI - Homeopathy in the paediatric population. PMID- 19675833 TI - [Not Available]. PMID- 19675834 TI - When your child needs a transfusion. PMID- 19675835 TI - [Not Available]. PMID- 19675836 TI - Paediatric research in Canada: What else can we do? PMID- 19675837 TI - IMPACT Highlights. PMID- 19675838 TI - Paediatric research in Canada: 2000-2004. PMID- 19675839 TI - Management of 21-hydroxylase deficiency congenital adrenal hyperplasia: A survey of Canadian paediatric endocrinologists. AB - BACKGROUND: Congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency is one of the most common autosomal recessive disorders. Many issues in the management of CAH in children still remain unresolved. OBJECTIVE: To assess how children with CAH are treated in Canada. METHODS: Fifty-nine paediatric endocrinologists and postgraduate trainees from across Canada took part in a survey that evaluated four areas of CAH management: type and dose of glucocorticoid therapy, current use of alternative therapies, monitoring of care, and approach/attitude to prenatal diagnosis and treatment of CAH. RESULTS AND CONCLUSIONS: The present survey demonstrated that there is general agreement among paediatric endocrinologists in Canada regarding the management of patients with CAH, which includes very little use of newer antiandrogen therapies. The goal remains to be the optimization of currently available therapy to ensure normal growth and sexual maturation without any evidence of glucocorticoid excess or deficiency. Prenatal diagnosis and management is widely, but infrequently, used. PMID- 19675840 TI - Nonsurgical correction of congenital ear abnormalities in the newborn: Case series. AB - OBJECTIVE: To determine whether a simple, nonsurgical treatment for congenital ear abnormalities (lop-ear, Stahl's ear, protruding ear, cryptotia) improved the appearance of ear abnormalities in newborns at six weeks of age. METHODS: This is a descriptive case series. All newborns with identified abnormalities were referred by their family physician to one paediatrician (WGS) in a small level 2 perinatal centre. The ears were waxed and taped in a standard manner within 10 days of birth. Pictures were taken before taping and at the end of taping (one month). All patients and pictures were assessed by one plastic surgeon (JWT) at six weeks of age and scored using a standard scoring system. A telephone survey of the nontreatment group was conducted. RESULTS: The total number of ears assessed was 90. Of this total, 69 ears were taped and fully evaluated in the study (77%). The refusal rate was 23%. In the treatment group, 59% had lop-ear, 19% had Stahl's ear, 17% had protruding ear and 3% had cryptotia. Overall correction (excellent/improved) for the treatment group was 90% (100% for lop ear, 100% for Stahl's ear, 67% for protruding ear and 0% for cryptotia). In the nontreatment (refusal) group, 67% of the ears failed to correct spontaneously. No complications were recognized by the authors or parents by six weeks. The percentage of newborns in one year in the perinatal centre with recognized ear abnormalities was 6% (90 of 1600). CONCLUSIONS: A simple, nonsurgical treatment in a Caucasian population appeared to be very effective in correcting congenital ear abnormalities with no complications and high patient/parent satisfaction. PMID- 19675841 TI - Disclosure of research results to research participants: A pilot study of the needs and attitudes of adolescents and parents. AB - BACKGROUND: Researchers have a moral responsibility to offer to return research results to participants, but the needs and attitudes of parents and adolescents with cancer in paediatric oncology regarding the issue are relatively unknown. OBJECTIVES: To explore the needs of potential research participants or their guardians with respect to the offer of a return of research results. METHODS: A questionnaire was used in a focus group and in telephone interviews with eight adolescents and 12 parents of children with cancer. The participants were asked to respond to the questions and to comment on the inclusiveness of the questionnaire. RESULTS: The majority of participants (18 of 20) wished to receive research results. Two somewhat unexpected findings are described. First, all participants in the present study felt that it was the primary responsibility of the participant to retain contact with the researchers for the purpose of obtaining research results. Second, few participants (n=2) indicated that the Internet would be a satisfactory way of transmitting these results. One-half of the participants wished to have face-to-face communication of results. CONCLUSIONS: These results provide preliminary guidance for the return of research results to participants and validate the use of the questionnaire in a larger study of this issue. PMID- 19675842 TI - Use of anencephalic newborns as organ donors. PMID- 19675843 TI - [Not Available]. PMID- 19675845 TI - When is my child ready for sports? PMID- 19675844 TI - Sport readiness in children and youth. PMID- 19675846 TI - [Not Available]. PMID- 19675847 TI - CPSP Highlights. PMID- 19675848 TI - We're listening: Members help inspire reflection at the Canadian Paediatric Society. PMID- 19675849 TI - The common postgraduate year 1: A paediatric perspective. PMID- 19675850 TI - Health promotion and research in the Chinese community in Montreal: A model of culturally appropriate health care. AB - In the past three decades, rapidly changing demographic characteristics have been witnessed in western society. Concurrently, numerous culture-related studies have attested to the disparity in health services among different ethnic groups. Reports have shown contributing factors that can render main stream services inaccessible to immigrants include cultural and language barriers, adaptation difficulties, racial discrimination and lack of culturally appropriate psychosocial services. Inadequate training of health professionals regarding cross-cultural issues may hinder patient compliance and therapeutic alliances. In a paediatric practice in Montreal, prevalent health problems among Chinese families were identified to be related to cultural beliefs and psychosocial factors. These included a low breastfeeding rate (8%) among Chinese newborns, prevalent iron deficiency anemia (12%), nursing caries, social isolation, delayed diagnosis of developmental delay, learning disabilities and psychosocial problems. The participation in community research and advocacy, the development of health promotional projects with an interdisciplinary approach and the preparation of Chinese health information such as perinatal programs, infant nutrition, dental hygiene and immunization, were described. Subsequent studies in this population illustrated an increased breastfeeding rate (48%), decreased incidence of iron deficiency (2.8%) and improved self-health maintenance practices. More culturally appropriate allied health services, including the Multiculturalism Department in the Montreal Children's Hospital, have subsequently been developed for the community.In summary, identifying prevalent health problems of ethnic propensity may be the initiatives needed to develop research and health promotional projects for a target population. Comprehensive health care to different ethnic groups requires health professionals to incorporate cross-cultural issues in their services. PMID- 19675851 TI - Circumcision policy: A psychosocial perspective. AB - The debate about the advisability of circumcision in English-speaking countries has typically focused on the potential health factors. The position statements of committees from national medical organizations are expected to be evidence-based; however, the contentiousness of the ongoing debate suggests that other factors are involved. Various potential factors related to psychology, sociology, religion and culture may also underlie policy decisions. These factors could affect the values and attitudes of medical committee members, the process of evaluating the medical literature and the medical literature itself. Although medical professionals highly value rationality, it can be difficult to conduct a rational and objective evaluation of an emotional and controversial topic such as circumcision. A negotiated compromise between polarized committee factions could introduce additional psychosocial factors. These possibilities are speculative, not conclusive. It is recommended that an open discussion of psychosocial factors take place and that the potential biases of committee members be recognized. PMID- 19675852 TI - Sudden death of a three-month-old infant while cosleeping on a sofa. PMID- 19675853 TI - Children at risk of developing amblyopia: When to refer for an eye examination. AB - Amblyopia is a sensory vision deficit that may be detected, diagnosed and treated best during the early stages of visual development. Only a small percentage of the paediatric population receive oculovisual examinations during this critical period of development even though such care is often free and easily accessible. Vision screening programs exist in certain states in the United States and in several European countries. Unfortunately, these programs are rarely implemented in Canada because of limited resources.Alternatively, screening at-risk infants may be more efficient and cost-effective in the early detection of amblyopia. Educating and training health care professionals, such as paediatricians, to recognize certain risk factors for amblyopia and encouraging early referral of the at-risk child to an eye care professional may prevent the development of amblyopia. The at-risk child has been defined to guide paediatricians and other primary health care professionals as to when to refer these children, the appropriate frequency of follow-up evaluations and the comanagement of compliance and treatment. PMID- 19675854 TI - Are acquired demyelinating syndromes of the central nervous system underdiagnosed in Canadian children? PMID- 19675855 TI - Recommendations for snowmobile safety. PMID- 19675856 TI - Head lice infestations: A clinical update. PMID- 19675858 TI - Creating a safe sleep environment for your baby. PMID- 19675857 TI - Recommendations for safe sleeping environments for infants and children. PMID- 19675859 TI - Can gallium-68 compounds partly replace (18)F-FDG in PET molecular imaging? AB - The development of gallium-68 -1,4,7,10-tetraazacyclodecane-1,4,7,10-tetraacetic acid ((68)Ga-DOTA) compounds was made possible due to the chemistry of (68)Ga, which matches the pharmacokinetics of many peptides, specially the chelators DOTA and DOTAderivatives with the formation of stable (68)Ga (3+) complexes. The availability of this tracer from a germanium-68-gallium-68 generator with a relatively long half-life makes it attractive to use in busy nuclear medicine departments, particularly those with limited access to cyclotrons. The recent clinical experience with (68)Ga-peptides includes imaging neuroendocrine tumours particularly carcinoid, as well as neuroectodermal tumours such as phaeochromocytoma and paraganglioma. In vitro and animal testing are still progressing alongside clinical studies, with promising results in the use of (68)Ga-DOTA-rhenium-cyclized alpha-melanocyte stimulating hormone (MSH) and (68)Ga-DOTA-napamide (NAP) in melanoma, (68)Ga-DOTA-PEG(4)-BN(7-14) (PESIN) for the imaging of bombesin receptor- positive tumours and (68)Ga-ethylene dicysteine metronidazole (EC-MN) for imaging tumour hypoxia. In addition to tumours, (68)Ga- DOTA peptide inhibitor of vascular peptide protein 1(VAP-P1) is being assessed for imaging inflammatory reaction. An additional value following a positive scan is the use of beta emitters labelled to the same peptides for radionuclide treatment. In conclusion, the recent introduction of (68)Ga-peptides, made available by a convenient (68)Ga/(68)Ge generator, could greatly contribute to the management of a wide range of clinical conditions including tumours and inflammation. PMID- 19675860 TI - Prostate cancer incidence, mortality, total and free prostate specific antigen. AB - The causes of prostate cancer (PC) and its progression are not yet known. Mortality and incidence from PC has increased throughout European countries until early 1990s. In Greece, PC is the second commonest cancer in men after lung cancer. The traditional Greek diet old in its origin may protect against common chronic diseases, including PC. Hormonal causes have been postulated in the aetiology of PC, mainly because androgen ablation causes regression of PC. Total and free prostate specific antigen (PSA and fPSA) screening were introduced to detect PC at an early stage, to reduce PC specific mortality, differentiate PC from hypertrophy and monitor response after hormonal treatment in advanced PC. PMID- 19675861 TI - Semi-quantitative analysis of perfusion of Brodmann areas in the differential diagnosis of cognitive impairment in Alzheimer's disease, fronto-temporal dementia and mild cognitive impairment. AB - Different perfusion defects reflect neurological damage characteristics of different kinds of dementia. Our aim was to investigate the role of brain single photon emission tomography (SPET) with semiquantitative analysis of Brodmann areas in dementia, by technetium-99m - hexamethyl-propylenamine- oxime ((99m)Tc HMPAO) brain SPET with semiquantitative analysis of Brodmann areas in patients with Alzheimer's disease (AD), frontotemporal dementia (FTD) and mild cognitive impairment (MCI). We studied 75 patients, 25 with AD (NiNCDS ADRDA criteria), 25 with FTD (Lund and Manchester criteria), 25 with MCI (EADC criteria). After i.v. injection of 740MBq of (99m)Tc-HMPAO, each patient underwent brain SPET. A software application was used able to map the SPET brain image to a stereotaxic atlas (Talairach), providing an affine co-registration by blocks of data defined in the Talairach space. A normal database calculating voxel by voxel the mean and the standard deviation of the measured values was built. Functional SPET data of 3D regions of interest (ROI) of predefined Brodmann's area templates were compared with those of a database of healthy subjects of the same age and gender. Mean values obtained in the Brodmann area ROI in the different groups of patients studied were evaluated. Our results showed that different Brodmann areas were significantly impaired in the different categories of dementia subjects. Both areas 37 (temporal gyrus) and 39 (angular gyrus) of AD patients (mean+/-SD: 37L= 1.6+/-1.0; 37R= -1.5+/-1.1; 39L= -2.3+/-1.3; 39R= -1.9+/-1.2) showed significant hypoperfusion (P<0.05) versus MCI (37L= -0.9 +/-0.7; 37R= -1.1+/-0.9; 39L= -1.4+/ 1.1; 39R= -1.6+/-1.6.) and FTD (37L= -1.1+/-0.8; 37R= -1.0+/-0.9; 39L= -1.4+/ 1.0; 39R= -1.2+/-1.2) subjects. AD patients showed significantly (P<0.01) decreased perfusion in areas 40 (supramarginal gyrus) (40L= -2.6+/-1.0; 40R= 2.3+/-1.1) with respect to MCI patients (40L= -1.8+/-0.9; 40R= -1.7+/-1.2). Finally, FTD patients showed significant hypoperfusion (P<0.05) in both areas 47 (frontal association cortex) (47L= -1.8+/-0.8; 47R= -1.1+/-0.8) in comparison with MCI subjects (47L= -1.2+/-0.9; 47R= -0.9+/-0.9). In conclusion, our results suggest that semiquantitative analysis of single Brodmann areas identify frontal area hypoperfusion in FTD patients and also parietal and temporal impairment in AD patients. In MCI patients, no hypoperfusion pattern is identified. PMID- 19675862 TI - Comparison of the effect of positive and negative oral contrast agents on (18)F FDG PET/CT scan. AB - Our aim was to compare the effect of orally taken 1% diatrizoate meglumine, 5% mannitol and water before positron emission tomography/computerized tomography (PET/CT) scan on gastrointestinal tract delineation and fluorine-18 fluorodeoxyglucose ((18)F-FDG) uptake. Our methods were as follows: Sixty-one patients referred for PET/CT scan without gastrointestinal diseases were divided into three groups. One thousand mL of 1% diatrizoate meglumine was orally taken 50 min before PET/CT scan in Group 1 (n=25), 1000 mL 2.5% mannitol was orally taken before scan in Group 2 (n=20) and 1000 mL water was orally taken before scan in Group 3 (n=16). Serum glucose and insulin were tested before and 45 min after taking mannitol in Group 2 patients. Paired t test was used to compare the glucose and insulin changes. The degree of gastrointestinal filling and (18)F-FDG uptake were evaluated by three nuclear medicine physicians using a 4 grade classification standard. Kruskal-Wallis and Mann- Whitney none parametric test was used to compare the filling condition and (18)F-FDG uptake difference among the three groups and between each group. RESULTS: the differences of serum glucose and insulin levels were not significant before and after contrast taken, in Group 2 patients. Group 2 patients had better gastrointestinal filling than patients of Group 1. Also, Group 2 patients' gastrointestinal filling was better than in Group 3 except in rectum. The jejunum, ascending, transverse and descending colon were better filled in Group 1 patients than in Group 3 patients. The degree of (18)FFDG uptake in stomach, jejunum and ileum, in Group 2 were significantly lower than those of Group 3 (P<0.05). (18)F-FDG uptake in jejunum, in Group 1 was also lower than in Group 3 (P<0.05). (18)F-FDG uptake in ascending colon in Group 1 was higher than in Group 3 (P<0.05). (18)F-FDG uptake in transverse and descending colon, in both Group 1 and Group 2 was significantly higher than in Group 3 (P<0.05). (18)F-FDG uptake in rectum, in Group 2 was significantly higher than in Group 3 (P<0.01). The average maximum CT values in stomach, jejunum, ileum and ascending colon in Group 1 patients were: 132+/-23, 191+/-31, 313+/-47 and 374+/-53 Hounsfield units respectively (Mean+/-SD, P<0.01 between every two groups). In conclusion, patients who take iso-osmia mannitol have good gastrointestinal filling, less physiological (18)F-FDG uptake and may thus have better (18)F-FDG images displaying gastrointestinal abnormalities and differentiating pathological from physiological lesions. PMID- 19675863 TI - Evaluation by (99m)Tc-pertechnetate scintigraphy of the effect of levocetirizine on salivary glands function, in allergic rhinitis patients. AB - The purpose of this study was to assess the effect of levocetirizine (xyzal) on salivary glands function in patients with allergic rhinitis using technetium-99m pertechnetate ((99m)Tc-P) salivary gland scintigraphy. The study population consisted of 67 patients with allergic rhinitis and 31 healthy controls (14 males and 17 females, mean age 30.1+/-6.8 years). The patients were divided into two groups: an untreated patient Group of 32 patients, 17 males and 15 females, mean age 29.9+/-6.5 years and a levocetirizine-treated with 5mg.day(-1) for 4 weeks patient Group, consisted of 35 patients, 16 males and 19 females, mean age 33.5+/ 7.8 years. All patients and healthy controls underwent salivary glands scintigraphy. After the intravenous administration of 185 MBq of (99m)Tc-P, dynamic salivary glands scintigraphy was performed for 25 min. By the time activity curves, the following glandular function parameters were calculated for the parotid and submandibular salivary glands: uptake ratio, maximum accumulation, and ejection fraction. Our results showed: All functional parameters obtained for the untreated patient Group were significantly lower than for the levocetirizine-treated patient Group and healthy controls (P<0.05). There was no significant difference in any functional parameters between the levocetirizine-treated patient Group and healthy controls (P>0.05). In conclusion, results of our study indicate that patients with allergic rhinitis treated with levocetirizine showed a significantly higher salivary glands function compared with untreated patients and healthy controls. Levocetirizine treatment showed no side effects on salivary glands function. PMID- 19675864 TI - A pilot study of changes in (18)F-FDG uptake, calcification and global metabolic activity of the aorta with aging. AB - Our aim was to quantify changes in the inflammatory and calcific components of atherosclerosis in the aortic wall using fluoro-18-2-fluoro-2-deoxy-D-glucose positron emission tomography (18)F-FDGPET and contrast enhanced computerized tomography (CECT) with increasing age. Twelve subjects, 8 men and 4 women aged from 21-80 years who had both (18)F-FDG-PET and CECT of the chest and abdomen were included in this study. Subjects were grouped into three according to age. (18)F-FDG uptake in four segments of the aorta was measured. Using CECT images, aortic segmental wall volumes were measured. Wall calcification volume in each aortic segment was also measured via adaptation of a coronary artery calcium scoring program to the aorta. Calcification volumes were then subtracted from aortic wall volumes. Each net segmental aortic wall volume was then multiplied by the accompanying mean SUV of the segment to calculate global metabolic activity (GMA) for each aortic segment. Our results showed that in each aortic wall segment, mean SUV, wall volumes, wall calcification volumes, and GMA statistically significantly increased with age. In conclusion, (18)F-FDG uptake, wall volume, wall calcification volume, and GMA in the aorta increase with aging. The (18)F-FDG uptake represents the early inflammatory component of the atherosclerotic process, whereas calcification generally represents a later and irreversible stage of the disease. Measurement and combination of PET and CECT parameters to calculate GMA may allow for optimal morphologic and functional noninvasive quantitative assessment of global aortic atherosclerotic disease. PMID- 19675865 TI - Effect of furosemide on radioiodine-131 retention in mice thyroid gland. AB - Retention of iodine in the thyroid gland is the result of renal excretion and transport of iodine to thyroid cells. Both processes are affected by furosemide. The aim of our study was to test whether furosemide influenced radioiodine-131 ((131)I) retention in the thyroid gland of living mice. Our methods were as follows: After 15 days of low-iodine diet, 19 Swiss mice received an intra peritoneal injection of 0.37+/-0.03 MBq of (131)I. Thereafter, 11 mice were treated with intraperitoneal injections of furosemide (0.3 mg/kg, every 8 h, for 72 h), Group A and 8 mice served as controls, Group B. Seventy-two hours after (131)I administration, the mice were anaesthetized, their thyroids were carefully extirpated, and their radioactivity was measured by a gamma counter. Our results showed that the mean value of (131)I retention after 72 h was 63.09% in Group A and 82.25% in Group B. The difference between these two groups was significant (T=3.0919, P=0.0033). In conclusion, furosemide after the administration of (131)I, decreases retention of (131)I in the thyroid gland in mice. The well known increase of iodine renal excretion by furosemide and consequently decrease of iodine blood pool may be the reason for this decreased (131)I retention by the thyroid gland. PMID- 19675866 TI - High accuracy of myocardial perfusion imaging in patients with left bundle branch block: comparison of four interpretation approaches. AB - Although myocardial perfusion imaging (MPI) with pharmacologic stress is the standard method for screening coronary artery disease (CAD) in patients with left bundle branch block (LBBB), controversies remain about its correct interpretation. We sought the best interpretation approach in these patients to achieve higher accuracy. Forty-two patients with LBBB underwent MPI with dipyridamole stress and the criteria for positive results with four patterns of interpretation were as follows: Pattern A: any reversible or irreversible perfusion abnormality in the myocardium irrespective of the location or extension was considered positive. Pattern B: any reversible or irreversible perfusion abnormalities except in the septal/anteroseptal region were defined as positive. Pattern C: in the absence of fixed LV cavity dilatation, the scan was interpreted the same as pattern A, while in the presence of fixed LV cavity dilatation, only the abnormalities outside the LAD territory was defined as positive. Pattern D: as in pattern C, except that in the absence of fixed LV cavity dilatation, the scan was read according to pattern B. For all patients, the angiographic results were considered as gold standard of CAD diagnosis. Our results showed that the false positive rate of MPI in patients with fixed LV dilatation was 50%, while in cases with normal LV size or transient dilatation, was 38.5%. This difference was more prominent in the female patients. The accuracy for screening CAD for patterns A, B, C and D were 57%, 62%, 69% and 69%, respectively. Pattern D was the better approach in female cases and patients with fixed septal/anteroseptal defects. In conclusion, a) in the male population without fixed defects in the septal/anteroseptal region, the specificity and accuracy are high in all patterns and the pattern of reading does not significantly influence the diagnostic value of MPI for CAD screening. b) in LBBB patients, fixed defects limited to the septal/anteroseptal region should be considered a significant finding only when LV cavity is not dilated. PMID- 19675867 TI - Gamma probe (99m)Tc-pertechnetate assisted completion thyroidectomy vs conventional thyroidectomy in differentiated thyroid carcinoma. AB - Patients undergoing partial thyroidectomy for benign diseases may need re operation if differentiated thyroid carcinoma (DTC) is detected on histopathology. The aim of this study was to determine if using gamma probe during the above surgery in a procedure called: gamma probe completion thyroidectomy (GPCT) could support the diagnosis of DTC tissue and offer an advantage in the surgical treatment of DTC patients. We have studied 100 patients who after bilateral subtotal thyroidectomy for benign disease in several hospitals, were found to have DTC histopathologically and referred to our clinic for subsequent re-operation. Of these, 50 underwent conventional completion thyroidectomy (Group I) and 50 underwent GPCT (Group II). We compared retrospectively Group I and Group II in terms of volume of residual thyroid tissue, thyroid stimulating hormone (TSH) values, complication rates and incidence of tumor found in the residual thyroid. Our results showed that one month postoperatively, TSH was significantly higher in Group II (P<0.001). Volumes of residual thyroid were also significantly less in Group II (P<0.000). Complications and the incidence of tumor cells found in the residual thyroid tissue between the groups were not statistically different (P>0.05). In conclusion, GPCT in patients with DTC significantly increased the success of this operation in localizing and removing residual thyroid tissue. PMID- 19675868 TI - Iotandium-111 pentetreotide scintigraphy and CT scans after 3 years in the follow up of patients with malignant melanoma. AB - Somatostatin receptor scintigraphy (SRS) has been used for the detection of neuroendocrine tumors. Melanoma is a malignant tumor of melanocytes, considered to derive from the neural crest. As the prognosis of melanoma is very poor, early detection of the disease, of recurrences and of distal metastases, is important. The aim of our study was to evaluate the clinical impact of indium-111 diaethyleno triamino pentaacetic acid-d-phe1-octreotide ((111)In-DTPA-octreotide or octreoscan or (111)In-O) in the management of melanoma patients after first diagnosis and first surgery and during three years of follow-up. We have studied 35 patients 20 female and 15 male, with histological proven melanoma. Scintigraphic images with single photon emission tomography gamma-camera (Millenium GE-USA) were performed after the administration of 220MBq (111)In-O. The scintigraphic data were compared to axial computerized tomography (CT). Patients were followed for 3 years after the initial diagnosis and surgery. Our results showed that during the 3 years follow-up period, 26/35 patients had a clinical recurrence. Twenty of them had positive (111)In-O scans with 56 lesions mainly metastatic, while 6 had negative scans. The CT scans showed only 31/56 lesions. In conclusion, SRS with (111)In-O, for diagnosing metastases from malignant melanoma, showed a sensitivity and specificity of 87% and 94% respectively and within the 3 years of follow-up, the stage of melanoma and surgical strategies were modified by 48% and 32%, respectively. Twenty five tumor sites, unsuspected by CT were visualized by (111)In-O. PMID- 19675870 TI - Can response to palliative treatment with radiopharmaceuticals be further enhanced? AB - Bone-seeking radiopharmaceuticals (RF) are an effective systemic pain-palliative treatment schedule in patients with disseminated bone disease. This treatment relies on selective uptake and prolonged retention of RF at sites of increased osteoblastic activity, while the potential toxicity of systemic administration is reduced by relatively selective tumour targeting. The main theoretic advantage of bone-targeted radionuclide treatment is that radiation can be delivered selectively to subclinical tumours and to metastases that are too small to be imaged and treated by surgical excision or local external beam radiotherapy. The distribution of radiation dose throughout a tumour is heterogeneous, whilst the rate of response depends upon the pre-treatment condition of the patient and other factors. To enhance the effect of treatment with RF on cancer cells, the following strategies have been investigated: Individualisation of the administered dose, higher-doses, earlier or repeated radionuclide treatment, the use of new RF, and the use of a mixture of different RF. Furthermore, external beam radiotherapy, biphosphonates or chemotherapeutic agents have been combined with RF and have scored significantly better treatment results referring to pain relief, bone disease progression, etc, without important side effects. The role of bone seeking RF can be extended beyond bone pain palliation, to a synergistic anti-tumour effect. PMID- 19675869 TI - TSH receptor antibodies for confirming the diagnosis and prediction of remission duration, in newly diagnosed Graves' disease patients. AB - Antibodies to TSH receptors (TRAb) have great pathogenetic importance in the development and maintenance of hyperthyroidism in Graves' disease (GD). Measurement of their serum level could be of diagnostic and prognostic importance in autoimmune hyperthyroidism and in monitoring the efficiency of thyrosuppressive drug (TSD) treatment in GD. The aim of our study was to assess the sensitivity (SN), specificity (SP), positive predictive value (PPV) and negative predictive value (NPV) of the TRAb assay in confirming the diagnosis and to define its prognostic value for remission duration in GD patients at the end of TSD treatment with thionamide derivatives, propylthiouracil or methimazole. We studied 99 patients, 65 female, 34 male, 18-76 years old; 52 GD patients and 47 as control group. We have studied frequency of relapses and remission duration in GD patients by a 2nd generation serum TRAb commercial kit. Our results showed that, the SN, SP, PPV and NPV of the TRAb test were 100%, 97%, 98% and 100%, respectively. Remission duration after TSD treatment was longer and relapses were fewer in GD patients with lower levels of TRAb before (P<0.01) and at the end of TSD treatment (P<0.05). In conclusion, our results suggest that serum TRAb is very sensitive and specific for confirming the diagnosis of GD. TRAb levels at the beginning of TSD treatment, above 5 IU/L gives 18% greater chance, and above 15 IU/L, 36% greater chance for remission, shorter than 6 months. PMID- 19675871 TI - Hepatobiliary scan in Alagille syndrome; arteriohepatic dysplasia. AB - Arteriohepatic dysplasia or congenital paucity of interlobular bile ducts - Alagille Syndrome, is a well defined syndrome characterized by five major features, including chronic cholestasis, posterior embryotoxon, butterfly-like vertebral arch defects, peripheral pulmonary artery hypoplasia or stenosis and facial dysmorphy. The disease is very rare. Only three cases have been reported in Greece and none with renal involvement. Hepatobiliary scan was a fundamental tool in the patients diagnosis and therefore we present the following case. PMID- 19675872 TI - A patient with medullary thyroid carcinoma and right ventricular cardiac metastasis treated by (90)Y-Dotatoc. AB - Medullary thyroid carcinoma (MTC) is rare derived from C cells of the thyroid gland and represents approximately 5% of all thyroid carcinomas. We report a case of a 74 years old male with MTC, diagnosed in 2002 and treated with total thyroidectomy and lymphadenectomy. A metastatic lesion was diagnosed on the right ventricle by indium-111-octreoscan, fluorine-18-fluorodeoxyglucose-positron emission tomography/computerized tomography, echocardiography, magnetic resonance imaging, high resolution computed tomography and was confirmed by histopathology. We present the results of treatment of this patient with yttrium-90-DOTA-tyr(3) octreotide. PMID- 19675873 TI - Imaging with (18)F-FDG-PET in infective endocarditis: promising role in difficult diagnosis and treatment monitoring. AB - Echocardiography (EC) plays a major role in diagnosing and monitoring the therapeutic response in infective endocarditis (IE) in routine practice. However in the setting of prosthetic valves or indwelling pacemakers, the EC findings are equivocal necessitating search for other diagnostic modalities. In these patients, (18)F-FDG/PET imaging may prove invaluable as evidenced by the presented case. We herein report a case of an 82 years old male with a mechanical aortic valve prosthesis who presented with a 10 days history of fever and malaise. Optimal interpretation of the EC results was difficult due to the presence of the prosthetic valve. However, (18)F-FDG-PET imaging findings were quite distinctive and revealed abnormally increased metabolic activity represented by two foci of increased (18)F-FDG uptake in the right and left borders of the heart that corresponded to areas of IE. PMID- 19675874 TI - Molecular imaging of carcinoid. PMID- 19675875 TI - (18)F-FDG-PET in restaging primary maxillary sinus melanoma with isolated gall bladder metastasis. PMID- 19675876 TI - Crossed cerebellar diaschisis demonstrated by (18)F- FDG-PET/CT. PMID- 19675877 TI - A nonpalpable toxic thyroid lobe besides a malignant lobe. PMID- 19675878 TI - Pitfall of (18)F-FDG-PET imaging in oncology: uterine fibromyoma. PMID- 19675879 TI - Uptake of (131)I on a post thyroid ablation whole body scan, due to cavernous liver hemangioma, mimicking metastases. PMID- 19675880 TI - Benign bone cyst mimicking skeletal metastasis on (18)F-FDG-PET. PMID- 19675881 TI - [Dosimetry in scintimammography by (99m)Tc-MIBI, (99m)Tc-Tetrofosmin, (99m)Tc (V)DMSA and 201TlCl compared with X-rays mammography]. AB - Scintimammography is sometimes referred as a complementary diagnostic method to X rays mammography and is the optimum examination for some patients to check suspected breast tumors. A dosimetric comparison of these diagnostic examinations is important. Estimation of the absorbed dose was achieved by calculations of internal dosimetry by Medical Internal Radiation Dose (MIRD) models and by calculations from the patients' scintigraphic images data using the MIRDOSE 3 program. The aim of this work was to compare the absorbed dose in scintimammography to that of X-rays mammography. Our results showed, that the breast absorbed dose by scintimammography examination using technetium-99m ((99m)Tc) complexes, 1.28mGy for (99m)Tc (V)-DMSA, 1.57mGy for (99m)Tc- MIBI and 1.92mGy for (99m)Tc-tetrofosmin, is considerably lower than the absorbed dose by scintimammography with thallium-201 chloride ((201)Tl-Cl) which reaches the 4.52mGy, or the absorbed dose by X-rays mammography. The absorbed mean glandular dose (MGD) for breast by X-rays mammography, taking 2 projections per breast, may reach 4.5mGy and is much higher for dense or large breasts. The European guidelines for quality assurance in breast cancer screening and diagnosis (2006, 4th edition) include a desirable level of less than 2.0mGy and an acceptable level of less than 2.5mGy for MGD for pressed breast thickness 4.5cm, per projection. In many radiology laboratories, specific care for every patient's radiation protection rules may reduce the MGD up to 1.4mGy per projection. Breast absorbed MGD in scintimammography by (99m)Tc-complexes mentioned above is reduced to more than half the above mentioned values for the examined breast and significantly lower than these levels for the not-examined breast. The radiation burden during scintimammography is also, many times lower than that of other radiology examinations including, lumbar spine radiography and pyelography. In conclusion, scintimammography is a low dose diagnostic method, and having also a high sensitivity, may be used as a complementary screening test in cases of dense breasts or breasts after surgery or breasts with prostheses, decreasing the necessity for breast biopsies. PMID- 19675882 TI - [Sodium-iodine symporter in thyroid, normal and cancer tissues and its relation to nuclear medicine and to gene cloning treatment]. AB - Sodium iodide symporter (NIS) is a transmembrane glucoprotein located in the basolateral membrane of the follicular thyroid cells, as well as in other normal and abnormal tissues such as the lactating mammary gland, well differentiated thyroid carcinoma and breast adenocarcinoma. It uses the electrochemical gradient generated by the Na-K ATPase to import an iodide molecule to the intracellular space along with two sodium molecules. The importance of NIS to diagnostic and research activities of Nuclear Medicine such as the radioiodine uptake, serum levels of TSH, TPO and TBG and thyroid diseases, especially cancer are described. NIS gene cloning in 1996 opened new prospective in diagnosis and treatment of thyroid and other diseases. PMID- 19675886 TI - Site specific protein labeling by enzymatic posttranslational modification. AB - Site specific protein labeling plays a key role in elucidating the function of the proteins at the molecular level by revealing their locations in the cell, their interaction networks with other cellular components and the dynamic mechanisms of their bio-generation, trafficking and degradation in response to regulatory signals in a biological system. Site specific protein labeling is, in essence, artificial modification of proteins with new chemical entities at the posttranslational stage. Based on the analogy between protein labeling and protein posttranslational modification, enzymatic tools have been developed for site specific and efficient labeling of target proteins with chemical probes of diverse structures and functionalities. This perspective surveys a number of protein labeling methods based on the application of protein posttranslational modification enzymes. PMID- 19675887 TI - High-yielding synthesis of Nefopam analogues (functionalized benzoxazocines) by sequential one-pot cascade operations. AB - An efficient amine-/ruthenium-catalyzed three-step process for the synthesis of Nefopam analogues was achieved through combinations of cascade enamine amination/iso-aromatization/allylation and diene or enyne metathesis as key steps starting from functionalized Hagemann's esters. In this communication, we discovered the application of ruthenium-catalysis on olefins containing free amines without in situ formation of salts. PMID- 19675888 TI - Enzymatic kinetic resolution of primary allenic alcohols. Application to the total synthesis and stereochemical assignment of striatisporolide A. AB - Crude Porcine pancreatic lipase was successfully used for the kinetic resolution of axially chiral primary allenic alcohols providing very high enantioselectivities with E values above 200. This simple access to optically active allenes was applied to the total synthesis of the fungal metabolite (-) striatisporolide A, allowing its unambiguous stereochemical assignment. PMID- 19675890 TI - Tandem SmI2-induced nitrone beta-elimination/aldol-type reaction. AB - Upon treatment with SmI(2), the carbohydrate-derived nitrones 1a,b undergo a beta elimination of the benzyloxy group at C-1, forming original samarium(III) oxy enamine intermediates. The latter can be reacted with carbonyl compounds to produce aldol-type adducts. The tandem process results in the transformation of a C-O bond into a C-C bond. PMID- 19675889 TI - Palladium-catalysed synthesis of 1-isoindolecarboxylic acid esters and sequential Diels-Alder reactions: access to bridged- and fused-ring heterocycles. AB - The Pd-catalysed intramolecular alpha-arylation of alpha-amino acid esters provides a useful methodology for the synthesis of substituted isoindole derivatives, which have been used in Diels-Alder reactions to access diverse skeletal frameworks. PMID- 19675891 TI - The influence of catechol structure on the suicide-inactivation of tyrosinase. AB - 3,6-Difluorocatechol, which cannot act as a monooxygenase tyrosinase substrate, is an oxidase substrate, and, in contrast to other catechols, oxidation does not lead to suicide-inactivation, providing experimental evidence for an inactivation mechanism involving reductive elimination of Cu(0) from the active site. PMID- 19675892 TI - Towards the conformational mimicry of the measles virus HNE loop: design, synthesis and biological evaluation of a cyclic bile acid-peptide conjugate. AB - The current article reports the design, synthesis and biochemical evaluation of a cyclic bile acid-peptide conjugate as a mimic of the loop-like structure of the measles virus haemagglutinin noose epitope (HNE). This macrocyclic structure was assembled by solid phase synthesis. Scaffold-peptide ring closure was achieved via the introduction of a succinate linker. After disulfide bridge formation with iodine, the desired 14 amino acid cyclic conjugate was obtained with overall yields between 15 and 35%. NMR analysis supports the presence of a helical conformation in the Q384-G388 pentapeptide portion, in agreement with the organisation of this chain in the native protein. The compound was found to have increased biostability compared to stabilised linear peptides, displayed good binding towards monoclonal antibodies known to bind to HNE and thus has potential in an alternative peptide-based measles vaccine. PMID- 19675893 TI - Practical synthesis of maleimides and coumarin-linked probes for protein and antibody labelling via reduction of native disulfides. AB - The cellular tracking, detection and sensing of protein or antibody movement are important aspects to advance our understanding of biomolecular interactions and activity. Antibodies modified with fluorescent dyes are also valuable tools, especially in immunology research. We describe here a proof-of-principle study of a new water-soluble coumarin probe with a maleimide thiol-reacting unit to fluorescently tag biomolecules. Highlights include: (1) a convenient water-based preparation of N-substituted maleimides, (2) a one-pot preparation of activated maleimido-esters, and (3) a bio-conjugation protocol for the selenol-promoted reduction of native disulfide bonds and the 'site-specific' labelling of antibodies with no significant loss of activity. PMID- 19675894 TI - Fluorescent substrates for covalent protein labeling catalyzed by microbial transglutaminase. AB - Novel small substrates with a variety of fluorophores were designed for the covalent labeling of proteins catalyzed by microbial transglutaminase (MTG). The new design is based on the flexibility in the substrate recognition of MTG for the substitution of the N-terminal protecting group of a conventional transglutaminase substrate, benzyloxycarbonyl-L-glutaminylglycine (Z-QG). Here we report for the first time that MTG can accept diverse fluorophores (dansyl, fluorescein, and rhodamine derivatives) in place of the benzyloxycarbonyl moiety when linked via a beta-alanine or epsilon-aminocaproic acid linker. The utility of the new fluorescent substrates was demonstrated by site-specific, covalent and quantitative labeling of an MTG-reactive Lys-containing peptide tag fused to the N-terminus of a recombinant bacterial alkaline phosphatase with retention of target protein functionality. PMID- 19675895 TI - A new class of conjugated strigolactone analogues with fluorescent properties: synthesis and biological activity. AB - A new class of strigolactone analogues has been synthesized. They differ from known molecules, both of natural and synthetic origin, in two main features. The conjugated system extends from the enol ether bridge to the A ring, the B ring is a heterocycle while the C ring is a cyclic ketone instead of a gamma-lactone. The key step of the synthesis is a Nazarov cyclization on activated substrates. Bioassays using Orobanche seeds have revealed that all the molecules strongly stimulate germination; in particular the oxygen containing analogues are the most active. Interestingly, some of the new molecules show fluorescent properties. PMID- 19675896 TI - Synthesis of 3-(1,2,3-triazol-1-yl)- and 3-(1,2,3-triazol-4-yl)-substituted pyrazolo[3,4-d]pyrimidin-4-amines via click chemistry: potential inhibitors of the Plasmodium falciparum PfPK7 protein kinase. AB - Efficient routes to 3-(1,2,3-triazol-1-yl)- and 3-(1,2,3-triazol-4 yl)pyrazolo[3,4-d]pyrimidin-4-amines using a one-pot two-step reaction are presented. The two routes give easy access to two different isomers of 1,4 disubstituted triazoles and the target compounds are obtained from a variety of readily available aromatic and aliphatic halides without isolation of potentially unstable organic azide intermediates. Two compounds show activity towards the PfPK7 kinase (IC(50) 10-20 microM) of P. falciparum, the organism responsible for the most virulent form of malaria, and can be regarded as hits useful for further development into lead compounds. PMID- 19675897 TI - High-yielding syntheses of hydrophilic conjugatable chlorins and bacteriochlorins. AB - Next-generation photodynamic therapy agents based upon the conjugation of multiple photosensitizers to a targeting backbone will allow for more efficacious light-based therapies. To this end, we have developed glucose-modified chlorins and bacteriochlorins featuring a reactive carboxylic acid linker for conjugation to targeting moieties. The photosensitizers were synthesized in relatively high yields from meso-tetra(p-aminophenyl)porphyrin, and resulted in neutral, hydrophilic chromophores with superb absorption profiles in the far-red and near infrared portions of the electromagnetic spectrum. In addition, conjugation of these photosensitizers to a model nanoscaffold (crosslinked dextran-coated nanoparticles) demonstrated that the inclusion of hydrophilic sugar moieties increased the number of dyes that can be loaded while maintaining suspension stability. The described compounds are expected to be particularly useful in the synthesis of a number of targeted nanotherapeutic systems. PMID- 19675898 TI - Enhancement of docetaxel solubility via conjugation of formulation-compatible moieties. AB - Computer-based theoretical calculations were employed to direct the design of docetaxel conjugates with enhanced solubility in the internal phase of a nano emulsion formulation. The theoretically-identified optimal docetaxel conjugates were synthesized by direct attachment of lauroyl moieties through an ester linkage to docetaxel. In comparison to docetaxel, the conjugates exhibited significantly improved solubility in oil, as predicted by our theoretical calculations. This contributed to high drug entrapment efficiencies (up to 97%) and a high drug loading capacity (5.7% w/w) for the docetaxel conjugates. The mono-substitution of an acyl group at C-2' of docetaxel resulted in a conjugate with 37- to 46-fold lower cytotoxicity than that of the parent drug in two human cancer cell lines. Importantly, the activity exerted by the mono-substituted docetaxel on the cancer cells was due in part to the cytotoxicity of the parent drug that was released via hydrolysis of the ester bond between the lauroyl moiety and the drug under biologically relevant conditions. In contrast, di- and tri-substitution of acyl groups at C-2', C-7 and/or C-10 of docetaxel resulted in non-hydrolysable conjugates that were found to be inactive. Overall, our results show that computer-based theoretical calculation is a promising strategy for guiding the enhancement of material-drug compatibility in formulation development. Also, these studies confirm that chemical modification of docetaxel for enhancement of material-drug compatibility should be limited to mono substitution at C-2' and result in a prodrug that is hydrolysable at a moderate rate under biologically relevant conditions. PMID- 19675900 TI - Studies towards the total synthesis of lycoposerramine A. Synthesis of a model for the tetracyclic core. AB - Lycoposerramine A (1) is a pentacyclic alkaloid isolated in 2001 by Takayama and co-workers. A concise synthesis of a model compound 8 for the tetracyclic core of this natural product is described. Key steps include the desymmetrising free radical cyclisation of compound 7 to give compound 18 and spirocyclisation of compound 26 to give compound 8. Earlier approaches using a novel high-yielding stereoselective anionic cyclisation of a cyclohexa-1,4-diene are also reported. PMID- 19675899 TI - Demonstration of bidirectional photoinduced electron transfer (PET) sensing in 4 amino-1,8-naphthalimide based thiourea anion sensors. AB - The thiourea based 4-amino-1,8-naphthalimide molecules 1-5 were designed as fluorescent anion sensors and their photophysical properties investigated upon recognition of biologically relevant anions such as acetate, dihydrogen phosphate and fluoride in DMSO. Synthesised in a single step from their respective aniline precursors, 6-9, these molecules were designed on the fluorophore-spacer-receptor principle, where in the case of sensors 1-3 the thiourea anion recognition moieties were connected to the fluorophore via the 4-amino moiety, while sensors 4 and 5 had the thiourea moieties connected to the 'imide' via a CH2 spacer. The current study showed that 1-5 operated as photoinduced electron transfer (PET) sensors, as no significant changes were observed in their absorption spectra, while their fluorescence emissions were quenched upon recognition of ions such as AcO(-), H2PO4(-) and F(-), which demonstrates that bidirectional PET sensing occurs in such naphthalimide based anion sensors. PMID- 19675901 TI - 8-Aza-2'-deoxyguanosine: base pairing, mismatch discrimination and nucleobase anion fluorescence sensing in single-stranded and duplex DNA. AB - Oligodeoxyribonucleotides containing 8-aza-2-deoxyguanosine 9 were synthesized and a new phosphoramidite 11, showing a high coupling yield in solid-phase synthesis, was prepared. Nucleoside 9 was found to be a perfect shape mimic of dG; it forms a strong base pair with dC and shows an excellent mismatch discrimination. Nucleoside 9 appears strongly fluorescent as the anion, and thus, it has unique reporter group properties as a replacement for dG. The pKa-value of 9 (8.4) is lower than that of dG (9.3) and increases from the single-stranded DNA (8.8) to duplex DNA (9.1). The fluorescence of the nucleoside 9 anion is decreased in ss-oligonucleotides and further reduced in duplex DNA. When mismatches of 9 are formed with the four DNA canonical bases, the fluorescence of mismatches is significantly higher than that of the perfectly paired duplex. The fluorescence of the nucleoside 9 anion correlates with the base pair stability. PMID- 19675902 TI - Acetylenic tetrathiafulvalene-dicyanovinyl donor-acceptor chromophores. AB - Compounds incorporating the tetrathiafulvalene (TTF) donor unit and one or two cyanoethynylethene (CEE) acceptor units were prepared by Knoevenagel condensations of highly unstable, TTF-linked propargylic aldehyde or ketone derivatives. The resulting TTF-CEEs are very strong chromophores with low-energy end-absorptions beyond 900 nm. The molecules experience reversible oxidations of the TTF unit, and the optical properties of the oxidised species were elucidated by spectroelectrochemistry. PMID- 19675903 TI - Synthesis and bioactivity of poly(HPMA)-lysozyme conjugates: the use of novel thiazolidine-2-thione coupling chemistry. AB - A novel thiazolidine-2-thione functionalized chain transfer agent (CTA) was synthesized and used as a reversible addition-fragmentation chain transfer (RAFT) polymerization agent to prepare well-defined poly-N-(2-hydroxypropyl) methacrylamide (PHPMA). The polymer chains had pre-designed molecular weights, narrow polydispersities and were chain-end functionalized. On incubation with protein (lysozyme) under different pH conditions, PHPMA was conjugated to the protein surface via covalent amide bonding. The bioactivity of the lysozyme-PHPMA conjugates was assessed using Micrococcus lysodeikticus (Ml) cells as substrates. The number of polymer chains attached to the protein could be controlled by both the pH of the conjugation reaction and the molecular weights of the polymers, thereby influencing significantly the bioactivity of the protein-polymer conjugates. PMID- 19675904 TI - Clickable fluorophores for biological labeling--with or without copper. AB - The synthesis of a set of new clickable fluorophores that virtually cover the whole visible spectrum reaching the near infra-red regime is presented herein. Besides dyes that are capable of participating in classical copper catalyzed 1,3 dipolar cycloaddition reactions with the counterparting function we have also prepared dyes containing a cyclooctyne moiety, an alkyne derivative that enables copper free clicking to azides. The suitability of these dyes for fluorescent labeling of biomolecules is presented by examples on model frameworks representing major biopolymer building blocks. The versatility of these dyes is presented in cell labeling experiments as well as by labeling the azide modified surface glycans of CHO-cells either by copper catalyzed or copper-free click reaction. These dyes are expected to have a large variety of applications in (bio)orthogonal labeling schemes both in vivo and in vitro. PMID- 19675905 TI - Phosphonate terminated PPH dendrimers: influence of pendant alkyl chains on the in vitro anti-HIV-1 properties. AB - The synthesis, characterization and in vitro anti-HIV activity of a series of generation one dendrimers having phosphonate groups with pendant alkyl chains are described. The influence of the lateral alkyl chains on the biological properties was correlated to (1)H-(1)H NOESY experiments. PMID- 19675906 TI - Fluoride recognition by a chiral urea receptor linked to a phthalimide chromophore. AB - The anion chemosensor 1 based on a urea-activated phthalimide with a stereogenic centre was synthesized using an efficient procedure involving a Curtius rearrangement. Its photophysical properties were estimated in several solvents. Sensor 1 detected fluoride with absorption as well as fluorescence changes and was only observable for this case and not for other halides. The appearance of a new CT complex emission at a longer wavelength and no changes in the singlet lifetime of 1 in the presence of fluoride supported a fluorescence static quenching mechanism. 1H-NMR studies, together with theoretical calculations based on DFT methods at the B3lYP/6-31G* level of theory confirmed the formation of a [1-F]- complex through H-bonding interactions rather than receptor deprotonation in the recognition process. Reversibility of this process was observed upon addition of a protic solvent. PMID- 19675907 TI - Anion-induced conformational changes in 2,7-disubstituted indole-based receptors. AB - The conformational preorganization and anion-induced conformational changes of indole-based receptors functionalized with an amide group at the 2-position and a variety of amide, urea and thiourea moieties at the 7-position have been studied by the means of NMR spectroscopy. NOE experiments showed that anti-anti orientation across C2-C2alpha and C7-N7alpha bonds is preferred for receptors 1-4 in acetone solution in the absence of anions. Anion-receptor interactions have been evaluated through (1)H and (15)N chemical shift changes. In 2,7-bis carboxamido functionalized indoles the interaction with chloride and bromide anions primarily occurs at the indole H1 proton. The introduction of urea and thiourea moieties increases the number of hydrogen bond donor sites which manifests itself in a distribution of halide-receptor interactions among the H1, H7alpha and H7gamma protons. Acetate anions also interact strongly with indole and urea NH donor groups, whereas nitrate anions interact solely with H7alpha and H7gamma urea/thiourea protons. NOE enhancements in the presence of anions revealed that anion-receptor complexes favour the syn-syn conformation of the C2 and C7 substituents. PMID- 19675908 TI - Easy access to trans-2,3-disubstituted cyclobutanones, 2,4,5-trisubstituted 3,6 dihydro-2H-pyrans and cis-substituted phenylcyclopropylsulfones by using the highly versatile 1-phenylsulfenyl- or 1-phenylsulfonyl-cyclopropylketones. AB - The high versatility of 1-phenylsulfenyl- or 1-phenylsulfonyl-cyclopropylketones has been exploited for the regioselective synthesis of trans-2,3-disubstituted cyclobutanones, 2,4,5-trisubstituted 3,6-dihydro-2H-pyrans and cis-2-alkyl- or cis-2-aryl-cyclopropylphenylsulfones. PMID- 19675909 TI - Application of carbodiimide mediated Lossen rearrangement for the synthesis of alpha-ureidopeptides and peptidyl ureas employing N-urethane alpha-amino/peptidyl hydroxamic acids. AB - Application of the Lossen rearrangement to the synthesis of N-urethane protected alpha-peptidyl ureas and ureidopeptides is reported. The carbodiimide mediated rearrangement of N-Boc/Z/Fmoc protected alpha-amino/peptide hydroxamic acids into isocyanates and coupling of the latter with the amino acid esters/peptide esters have been accomplished in a single-pot to obtain good yields of urea products. Synthesis of the ureidoalanine derivatives via the hydroxamate derivatives of N protected aspartic acid has also been carried out using the same procedure. PMID- 19675910 TI - Cycloaddition and one-carbon homologation studies in the synthesis of advanced iridoid precursors. AB - A Diels-Alder cycloaddition approach to the sweroside aglycone intermediate of iridoids was explored using silylated butenolides and levoglucosenone as dienophiles under both Lewis acid and thermal conditions. Results of this study reveal no evidence that using less sterically demanding derivatives compromise the diastereofacial selectivity of the cycloaddition using silylated butenolides. Further chemistry performed on cycloadducts concentrated on the identification and management of methodologies suitable for its conversion into sweroside aglycone. During the course of these studies, a dehydrative cyclisation onto a preformed tetrahydrofuran ring to a bis-tetrahydrofuranoid moiety was unravelled. In addition studies on levoglucosenone-derived cycloadducts provide extensive insight into the conformational behaviour and reactivity. Further, the X-ray crystal structure of an alcohol intermediate from one-carbon homologation studies provided the first structural evidence confirming the diastereoselectivity of the cycloaddition procedure. PMID- 19675912 TI - Rigid rod and tetrahedral hybrid compounds featuring nucleobase and nucleoside end-capped structures. AB - Being aimed at a new type of porous solids, a moduled design strategy of molecular tectons, making use of the conjugation between a shape defined artificial backbone and the bioinspired molecular fragments of nucleobases or nucleobase derivatives as functional end-caps, has been developed. This led to the formation of the new hybrid compounds 1-13 of linear and tetrahedral geometry, containing uracil, adenine, adenosine, guanosine and its acylated analogs as the sticky end-cap sites. The compounds were synthesized from a halogen or ethynyl substituted nucleobase component and the corresponding ethynylated spacer unit following a metal assisted coupling process as the key reaction step. X-Ray crystal structure analysis demonstrates that the parent compound 1 is a solvent complex with DMSO (1:2), showing the DMSO molecules incorporated in a hydrogen bonded layer structure. Specific dependencies of the fluorescence properties of the new compounds in solution on the structure of the molecules are reported. A selection of solid compounds has been studied in respect of their ability to adsorb organic vapours. They revealed significant differences both in the sorption capacity and the selectivity towards particular solvent vapours. PMID- 19675911 TI - A temporary stereocentre approach for the asymmetric synthesis of chiral cyclopropane-carboxaldehydes. AB - A novel way of combining chiral auxiliaries and substrate directable reactions is described that employs a three-step sequence of aldol/cyclopropanation/retro aldol reactions for the asymmetric synthesis of enantiopure cyclopropane carboxaldehydes. In the first step, reaction of the boron enolate of (S)-N propionyl-5,5-dimethyl-oxazolidin-2-one with a series of alpha,beta-unsaturated aldehydes affords their corresponding syn-aldol products in high de. In the second step, directed cyclopropanation of the alkene functionalities of these syn aldols occurs under the stereodirecting effect of their 'temporary'beta-hydroxyl stereocentres to give a series of cyclopropyl-aldols in high de. Finally, retro aldol cleavage of the lithium alkoxide of these cyclopropyl-aldols results in destruction of their temporary beta-hydroxy stereocentres to afford the parent chiral auxiliary and chiral cyclopropane-carboxaldehydes in >95% ee. The potential of this methodology has been demonstrated for the asymmetric synthesis of the cyclopropane containing natural product cascarillic acid in good yield. PMID- 19675913 TI - An investigation into the electrophilic cyclisation of N-acyl-pyrrolidinium ions: a facile synthesis of pyrrolo-tetrahydroisoquinolones and pyrrolo-benzazepinones. AB - The triflic acid-mediated cyclisation of N-arylmethyl- and N-arylethyl acylpyrrolidinium ions gave moderate to good yields of pyrrolo tetrahydroisoquinolones and pyrrolo-benzazepinones respectively. Electron donating R substituents enhanced the rate of reaction and gave higher yields than electron-withdrawing substituents. Substituents on the methyl or ethyl chain in general enhanced the reaction, unless sterically encumbered. The equivalent acylpiperidinium ions cyclised much slower and in lower yield. PMID- 19675914 TI - Solvent reorganisation as the driving force for rate changes of Menschutkin reactions in an ionic liquid. AB - The effect on the rate of reaction of each of a series of Menschutkin processes on changing from a molecular solvent to an ionic liquid was investigated. In each case, the rate acceleration observed at room temperature could be attributed to the change in the entropy of the system on reaching the transition state, offsetting any enthalpic cost. PMID- 19675915 TI - Understanding the mechanism of polar Diels-Alder reactions. AB - A good correlation between the activation energy and the polar character of Diels Alder reactions measured as the charge transfer at the transition state structure has been found. This electronic parameter controls the reaction rate to an even greater extent than other recognized structural features. The proposed polar mechanism, which is characterized by the electrophilic/nucleophilic interactions at the transition state structure, can be easily predicted by analyzing the electrophilicity/nucleophilicity indices defined within the conceptual density functional theory. Due to the significance of the polarity of the reaction, Diels Alder reactions should be classified as non-polar (N), polar (P), and ionic (I). PMID- 19675916 TI - The enantioselective benzoin condensation promoted by chiral triazolium precatalysts: stereochemical control via hydrogen bonding. AB - The design of a new class of triazolium ion precatalysts incorporating protic substituents is described. These materials promote the enantioselective benzoin condensation of a range of aromatic aldehydes (1-62% ee). Catalyst evaluation studies strongly support the involvement of hydrogen bond donation by the catalyst in the stereocentre-forming step of the catalytic cycle. PMID- 19675917 TI - Synthesis of functionalized Morita-Baylis-Hillman adducts by a conjugate addition elimination sequence. AB - We have developed a new conjugate addition-elimination sequence for the diastereoselective synthesis of protected allylic syn 1,3-diols which are Morita Baylis-Hillman adducts. The synthesis of the substrates involves a challenging cross-metathesis reaction that leads to hindered trisubstituted olefins. PMID- 19675918 TI - Enantioselective electrophilic trifluoromethylation of beta-keto esters with Umemoto reagents induced by chiral nonracemic guanidines. AB - Chiral nonracemic guanidines act as Bronsted bases to generate guanidinium enolates for the enantioselective electrophilic trifluoromethylation of beta-keto esters by means of S-(trifluoromethyl)dibenzothiophenium tetrafluoroborate (Umemoto reagent) with good enantioselectivity of 60-70% range. Despite the fact that the ees are still improvable, the model reported in this work could spark the imagination of chemists to design new chiral bases to improve the stereochemical outcome. PMID- 19675919 TI - Light driven formation of a supramolecular system with three CB[8]s locked between redox-active Ru(bpy)3 complexes. AB - Three CB[8]s have been reversibly locked between two Ru(bpy)(3)-viologen complexes by light driven electron transfer reactions. PMID- 19675923 TI - Short-term community dynamics in seasonal and hyperseasonal cerrados. AB - In South America, the largest seasonal savanna region is the Brazilian cerrado. Our aim was to study temporal changes in some community descriptors, such as floristic composition, richness, species density, plant density, and cylindrical volume, in a seasonal cerrado, comparing it to a nearby hyperseasonal cerrado. In four different seasons, we placed randomly ten 1 m(2) quadrats in each vegetation form and sampled all the vascular plants. Seasonal changes in floristic composition, species density, and plant density were less pronounced in the seasonal than in the hyperseasonal cerrado. Floristic similarity between the vegetation forms was lower when the hyperseasonal cerrado was waterlogged. Richness and species density were higher in the seasonal cerrado, which reached its biomass peak at mid rainy season. The hyperseasonal cerrado, in turn, reached its biomass peak at early rainy season and, despite the waterlogging, maintained it until late rainy season. In the hyperseasonal cerrado, waterlogging acts as an environmental filter restricting the number of cerrado species able to withstand it. The seasonal cerrado community was more stable than the hyperseasonal one. Our results corroborated the idea that changes in the environmental filters will affect floristic composition and community structure in savannas. PMID- 19675924 TI - Phytoplankton community: indicator of water quality in the Armando Ribeiro Goncalves reservoir and Pataxo channel, Rio Grande do Norte, Brazil. AB - The current study analysed spatial-temporal modifications of the phytoplankton community and water quality, during dry and wet seasons. The phytoplankton community was studied in three areas: Armando Ribeiro Goncalves Reservoir (ARG), which is an important public use reservoir in RN, Pataxo Channel (PC-before water treatment), Itaja, RN, and after the water treatment (WTP). Water samples from the reservoir were collected during both dry (January, February and November, 2006) and wet seasons (March to June, 2006). Quali-quantitative analyses of phytoplankton were carried out. Results indicated a qualitative similarity of the phytoplankton community in the three areas. However, significant differences were registered in these areas in relation to species relative abundance, with dominance of potentially toxic cyanobacteria, such as Planktothrix agardhii Gomont (dry season) and Microcystis aeruginosa Kutz (wet season). Ecological indexes obtained higher values before water treatment. Nevertheless, densities of cyanobacteria (organisms/mL) gradually reduced in the waters of the reservoir and of the Pataxo Channel before and after water treatment. After the treatment, density values of cyanobacteria were adequate for human consumption, according to the values established by the Health Ministry. PMID- 19675925 TI - Composition and distribution of Darwinulidae (Crustacea, Ostracoda) in the alluvial valley of the upper Parana River, Brazil. AB - The occurrence and abundance of darwinulid ostracods, as well as environmental factors influencing these patterns, were investigated in the alluvial valley of the upper Parana River. Ostracods were sampled from several substrates, like littoral sediments and pleuston, which included several aquatic macrophytes species, from 31 localities (lentic and lotic) belonging to different riverine systems. Eight darwinulid species were found, representing all genera from this family. Alicenula serricaudata, Vestalenula pagliolii, and Penthesilenula brasiliensis were the most common species. Cluster analysis based on the composition and abundance of darwinulid communities revealed the presence of five associations. Darwinula stevensoni, Vestalenula botocuda, and Penthesilenula aotearoa were almost exclusive to lotic environments. A Mantel multiple test showed that the occurrence and distribution of darwinulid ostracods were significantly related to types of habitat and systems, but not to abiotic variables. It thus seems that the hydrodynamic fluctuations of these environments are probably more important to darwinulid distribution than the limnological characteristics. PMID- 19675926 TI - Effects of the pollution by petroleum on the tracheids along the stem of Podocarpus lambertii Klotzsch ex Endl., Podocarpaceae. AB - Podocarpus lambertii Klotzsch ex Endl. (Podocarpaceae) is native and a member of the Pinophyta (Gymnosperm) of southern Brazil, locally known as 'pinheiro-bravo'. The present work aims to investigate the effects of petroleum on the tracheids dimensions. Wood samples from twenty individuals were studied along the stem, ten being exposed to pollution and ten used as a control set. The wood samples were collected from incisions at three levels: at the ground level, and one and two metres above the ground level. From these samples, sub-samples were selected at the border of the growth layers in the vascular cambium-medulla direction. The methodology followed that traditionally recommended for plant anatomy studies, with analyses done by light microscopy (OLYMPUS - BX41) assisted by the software Image Pro-plus for measurements. Comparison of the individuals exposed to petroleum with the control set, showed that the length, diameter and cell wall width of the tracheids of the former were smaller, a trend which was statistically significant according to the Student's t-test. These traits were observed mainly on the tracheids of the last growth layer, corresponding to the year in which the individuals were exposed to petroleum. PMID- 19675927 TI - Determination of eutrophic areas in Mundau/Manguaba lagoons, Alagoas-Brazil, through studies of the phytoplanktonic community. AB - Mundau and Manguaba Lagoons (9 degrees 34' 38'-9 degrees 45' 30' S and 35 degrees 44' 00'-35 degrees 58' 13' W) are considered the largest and most productive ones in the state of Alagoas and were studied with the purpose of identifying the existence of anthropic impacts. Samples were collected at 8 stations, during low tide and flooding in rainy and dry periods, using the 'Van Dorn' bottle and plankton net. In total, 155 taxons were identified, with special emphasis on Bacillariophyta and Cyanophyta divisions. The most abundant species in Mundau Lagoon was Skeletonema cf. costatum, and in the Manguaba Lagoon, Cyclotella meneghiniana, Microcystis aeruginosa and Anabaena spiroides. The diversity ranged from 0.17 bits cell/L to 4.81 bits cell/L. The predominance of freshwater species (51%), evidenced higher influence of the limnetic flow on the two environments studied. The high values related to the phytoplanktonic density characterize the lagoons as strongly impacted environments, indicating the existence of eutrophic conditions in most of the studied stations. PMID- 19675928 TI - Seasonal dynamics of methane in the water column of two subtropical lakes differing in trophic status. AB - Alterations in methane concentration in the water column of aquatic systems is closely linked to the processes of production and consumption of this gas, i.e., methanogenesis and methanotrophy respectively. The aim of this research is to evaluate methane dynamics through diurnal variation in the concentration of this gas in the water column of two lakes differing in trophic status at the campus of Fundacao Universidade do Rio Grande (FURG). In two sampling periods (November 2001 and July 2002) methane concentrations in the water column were significantly higher (3.66 and 0.41 micromoles.L-1, respectively) at Lago dos Biguas, with mesoeutrophic features when related to Lago Polegar (1.43 and 0.19 micromoles.L 1, respectively) which has oligotrophic features. The higher methane concentrations were detected in November 2001 when higher temperature was also detected. The results highlighted the importance of trophic status as well as seasonality for the methane dynamics in these ecosystems. PMID- 19675929 TI - Structure of the herb stratum under different light regimes in the Submontane Atlantic Rain Forest. AB - This study aimed to characterize the structure of the herb stratum in relation to light availability in the Submontane Atlantic Rain Forest at the Carlos Botelho State Park, SP, Brazil. Fortyone 10 x10 m plots were established under the closed canopy (18 plots), small and medium canopy gaps (11) and large canopy gaps dominated by Guadua tagoara (Ness) Kunth (12). Inside each plot, the line intercept method was applied to assess soil coverage as an estimate of density of herb stratum vegetation. Hemispherical photographs were taken at the centre of the plots to evaluate the annual light regime. Overall, Calathea communis Wanderley and S. Vieira had the greater mean coverage, followed by woody seedlings, ground ferns and other herbs (mainly, Araceae, Acanthaceae, Amaranthaceae and Cyperaceae). There were strong correlations among several groups of the herb stratum, such as the negative correlations between woody seedlings with the coverage of C. communis and with rocks. The analysis of the hemispherical photographs confirmed the difference among environments that led to significant differences in the soil coverage of the herb stratum vegetation but woody seedlings. For instance, C. communis showed great coverage in large gaps while ferns were more abundant in small and medium gaps and in the understorey. Other herbs, in turn, demonstrated bigger soil coverage in small and medium gaps. Although this study represents a rough assessment of the structure and composition of the herb stratum, the results found here illustrated the evident relation between herb species density and the environmental variation promoted by changes on canopy structure and topography. PMID- 19675930 TI - Ecology of endoparasites of the fluvial stingray Potamotrygon falkneri (Chondrichthyes: Potamotrygonidae) from the upper Parana River floodplain, Brazil. AB - The present study investigated the ecological relationships between endoparasites and the host Potamotrygon falkneri from the upper Parana River by using as indicators the host's relative condition factor (Kn), sex and hepatosomatic relation (HSR). Forty-seven specimens of P. falkneri were analyzed between March 2005 and September 2006. Statistical analysis showed that the Kn was positively correlated with the abundance of Acanthobothrium regoi and Rhinebothrium paratrygoni; only the abundance of A. regoi was positively correlated to the HSR, whereas R. paratrygoni did not present correlation and there was no influence of the host's sex on the abundance and prevalence of parasites. This is the first study concerning the ecology of parasites of potamotrygonids. PMID- 19675931 TI - Small mammals in a fragment and adjacent matrix in southeastern Brazil. AB - Between May 2002 and May 2003, we studied a small mammal community from an Atlantic forest fragment surrounded by a coffee plantation in the municipality of Santa Teresa, Espirito Santo state, Brazil. We obtained a total of 300 captures of 114 individuals belonging to 11 mammal species. Seventy three percent of the species captured in the forest also used the coffee plantation, mainly males (90%) and young (80%) individuals of Marmosops incanus. The exceptionally low recapture rate in the coffee plantation suggests that it functions as a corridor between fragments, rather than as a habitat for these species. Understanding the role of matrix habitat use in fragmented landscapes is an important factor in small mammal conservation, as it asymmetrically affects the rate and mode of individual movements of different species. PMID- 19675932 TI - Body size of commom opossum Didelphis aurita Wied-Neuwied 1826 (Didelphimorphia: Didelphidae) on southern brazilian islands. AB - The body size of vertebrates isolated on islands can undergo changes due to ecological features of these environments. This study aimed to compare the body size of the common opossum, Didelphis aurita, from different insular populations within the same archipelago in southern Brazil. The opossum populations showed corporal variation and different hypotheses were raised to understand the results. This study constitutes the most detailed body size comparison of a marsupial within different insular populations in the Neotropical zone and the data gathered represents an initial contribution for regional fauna biometric knowledge. PMID- 19675933 TI - Enzymes of energy metabolism in hatchlings of amazonian freshwater turtles (Testudines, Podocnemididae). AB - The metabolic profiles of selected tissues were analyzed in hatchlings of the Amazonian freshwater turtles Podocnemis expansa, P. unifilis and P. sextuberculata. Metabolic design in these species was judged based on the key enzymes of energy metabolism, with special emphasis on carbohydrate, lipid, amino acid and ketone body metabolism. All species showed a high glycolytic potential in all sampled tissues. Based on low levels of hexokinase, glycogen may be an important fuel for these species. The high lactate dehydrogenase activity in the liver may play a significant role in carbohydrate catabolism, possibly during diving. Oxidative metabolism in P. sextuberculata appears to be designed for the use of lipids, amino acids and ketone bodies. The maximal activities of 3 hydroxyacyl-CoA dehydrogenase, malate dehydrogenase, glutamine dehydrogenase, alanine aminotransferase and succinyl-CoA keto transferase display high aerobic potential, especially in muscle and liver tissues of this species. Although amino acids and ketone bodies may be important fuels for oxidative metabolism, carbohydrates and lipids are the major fuels used by P. expansa and P. unifilis. Our results are consistent with the food habits and lifestyle of Amazonian freshwater turtles. The metabolic design, based on enzyme activities, suggests that hatchlings of P. unifilis and P. expansa are predominately herbivorous, whereas P. sextuberculata rely on a mixed diet of animal matter and vegetation. PMID- 19675934 TI - A study of low power laser on the regenerative process of Girardia tigrina (Girard,1850) (Turbellaria; Tricladida; Dugesiidae). AB - The mechanism of regeneration does not start to restore the wound until its corresponding epimorphic phase. A bioestimulation of tissues and cells by laser radiation depends on the wavelength, on the dose, and on the intensity of the light. The goal of this work was to verify the effect of the low power laser at 660 nm on the regenerative process of Girardia tigrina. The specimens were maintained in the laboratory under a temperature ranging from 19 degrees up to 24 degrees C for 21 days. The planarians were anesthetized by placing them on ice and then cut them with a scalpel. The three treatments were as following: animals individually irradiated with 14 sessions with 1 minute duration (treatment 1), 14 sessions with 3 minutes duration (treatment 2), and without irradiation (control). The planarians were amputated and divided in three study treatments: a control group (without radiation), and two other treatments: irradiated for 1 minute, and irradiated for 3 minutes. The animals were irradiated with diode laser (660 nm) with 3.3 +/- 0.3 mW of power, using 0.94 mW.mm-2 power density for each irradiation procedure. During the experiment, 14 irradiation sessions were undertaken. The specimens were fixed in Bouin, and stained with hematoxyline and eosin. From observation and histological analysis, it was possible to assess the effects of interaction between laser and tissue. The head fragment after 1 minute of irradiation presented a better organized tissue scheme, when compared with the other treatments. Aspects of the body fragments submitted to 3 minutes of light treatment were very similar to fragments that had not been injured. It can be concluded that there are changes in the quality of regeneration when treated with low power laser under the conditions mentioned above. PMID- 19675935 TI - Effects of bacterial infestation caused by human wastes on the skin structures of Mugil platanus Gunther, 1880 (Mugilidae). AB - Manifestation of infectious pathologies in fishes usually increases in environments where organic wastes are disposed. Specimens of Mugil platanus Gunther, 1880 and water samples collected at three points of the Tramandai river were analyzed during a one year period. The macroscopic observation revealed ulcerations in the caudal peduncle area covered with a mass of amorphous and whitened tissues. Histopathologic analysis showed the presence of negative gram bacteria, probably responsible for alterations of the normal structure of the epidermic tissues. Non-parametric statistical analysis for ammonia concentration showed a significant variation among the three collected spots as well as in the multiple comparison between two spots. In this study, we describe cutaneous lesions observed in Mugil platanus specimens and tested their correlation with environmental ammonia concentration. PMID- 19675936 TI - Chemical variation of tannins and triterpenes in Brazilian populations of Maytenus ilicifolia Mart. Ex Reiss. AB - Maytenus ilicifolia and Maytenus aquifolia species are widely used as a cicatrisation agent, an anti-spasmodic, contraceptive, antiulcerogenic, diuretic and analgesic. Considering the importance of these species in popular medicine, this work is focused on the determination of the chemical content of tannins and the triterpenes friedelan-3-one, friedelan-3-ol and friedelin in 15 native populations of Maytenus ilicifolia distributed in the south and mind-west regions of Brazil. Correlation of the concentration of these compounds with the environmental parameters such as average annual temperature, climate, vegetation, geomorphology, latitude and altitude was determined using Pearson's coefficient. Results showed that average annual temperature and climate have significant effect on tannin content at a 95% confidence level. The highest tannin concentration was found in Ponta Pora population, and for the triterpenes investigated, a significant correlation between their concentrations with the environmental variables studied was not verified. PMID- 19675937 TI - Sexual dimorphism in the mandible of the armadillo Chaetophractus villosus (Desmarest, 1804) (Dasypodidae) from northern Patagonia, Argentina. AB - The aim of this study was to study the sexual dimorphism in adult Chaetophractus villosus (Desmarest, 1804), from northern Patagonia, Argentina. Eight mandibular traits were measured in 37 males and 34 females. Univariate and multivariate morphometric analysis were applied to the data set. Results showed that C. villosus was sexually dimorphic, with higher absolute values corresponding to females. The total length of the mandible was the most important variable to discriminate sexes, followed by the height at the level of the last tooth and body length. The percentages of sex discrimination were high, as they were when a new sample (17 males, 13 females) was tested. Females have larger mandibles than males independently of their larger cranial size. They also showed a higher degree of correlation between variables, suggesting a more stable shape for the mandible than in males. PMID- 19675938 TI - Use of palmae wax hydrocarbon fractions as chemotaxonomical markers in Butia and Syagrus. AB - The wax hydrocarbon fractions of native Butia and Syagrus species collected from Palms in different regions of the of Rio Grande do Sul state (Brazil) and in Rocha (Uruguay) were analyzed to evaluate their potential as chemotaxonomic markers. The wax was extracted with chloroform and the resulting wax was fractionated by preparative TLC. The hydrocarbon fractions were analyzed by GC MS. Statistical analyses were completed with the Statistica 5.0 program. The total crude wax yields averaged 0.31% w.w-1 dried leaves for Butia samples and 0.28% for Syagrus samples. The linear hydrocarbons represented on average 15% of the total waxes in the case of Butia samples and 13.7% in Syagrus samples. Hentriacontane and triacontane were the main components of all samples. The comparison of the means showed significant differences among Butia and Syagrus samples, and amongst Butia samples collected in different localities. In the case of the Syagrus collections no consistent groupings could be made. In the case of Butia samples the formation of three groupings could be observed, which were consistent with the species described for their geographical distribution. These results are discussed in the paper. PMID- 19675939 TI - Parasitoid diversity (Hymenoptera: Braconidae and Figitidae) on frugivorous larvae (Diptera: Tephritidae and Lonchaeidae) at Adolpho Ducke Forest reserve, central amazon region, Manaus, Brazil. AB - This study aimed to identify parasitoid species of frugivorous larvae and to describe the tritrophic interactions involving wild fruits, frugivorous insects and their natural enemies at Adolpho Ducke Forest Reserve (RFAD) (Manaus, AM, Brazil). Collections were performed in four 1 km2 quadrants in the corners of the RFAD. The wild fruits were collected inside the forest in access trails leading to each collection area and in trails that surrounded the quadrants, up to five metres from the trail on each side. The fruits were placed in plastic containers covered with thin fabric, with a vermiculite layer on the base to allow the emergence of flies or parasitoids. Seven Braconidae species were collected, distributed among Opiinae: Doryctobracon areolatus (Szepligeti, 1911), Utetes anastrephae (Viereck, 1913), and Opius sp., and Alysiinae: Asobara anastrephae (Muesebeck, 1958), Phaenocarpa pericarpa Wharton and Carrejo, 1999, Idiasta delicata Papp, 1969, and Asobara sp. Parasitism rates by braconids and figitids are presented. Doryctobracon areolatus was the most frequent, parasitizing the highest number of fly species, and showing the highest parasitism percentage in larvae feeding on Micropholis williamii fruits. The collected figitids belong to Aganaspis nordlanderi Wharton, 1998 and A. pelleranoi (Brethes, 1924). All 15 tritrophic associations are new records for the Brazilian Amazon region. The RFAD is an important natural reservoir of frugivorous larvae parasitoids. PMID- 19675940 TI - Steviol effect, a glycoside of Stevia rebaudiana, on glucose clearances in rats. AB - Stevia rebaudiana, a South American plant normally used as a natural herbal sweetener, has been suggested as exerting beneficial effects on human health, including as an antihypertensive and antihyperglycemic. The present experiment was undertaken to evaluate the renal excretion of steviol, the aglycone of several natural products extracted from the leaves of S. rebaudiana, and to clarify the actual participation of this compound on the renal excretion of glucose in rats, which has been previously suggested as the preferential action of steviol on the Na+-glucose renal tubular transport system. Steviol was obtained by enzymatic hydrolysis of stevioside with pectinase. Thirty normal male Wistar rats weighing 345 g were used. After a control period, steviol was infused iv at three doses (0.5, 1.0 and 3.0 mg.kg-1/h), according to classical clearance techniques. During all the experiments no significant changes in inulin clearance (Cin) and p-aminohipuric acid clearance (C PAH) were observed. Administration of steviol resulted in a statistically significant increase in the fractional sodium excretion (FeNa+), fractional potassium excretion (FeK+), urinary flow as percent of glomerular filtration rate (V/GFR) and glucose clearance (C G) when compared to controls, but these effects were absent with the dose of 0.5 mg.kg-1/h. The steviol clearance (C S) was higher than the Cin and lower than the C PAH at all the doses employed in this study. The data suggest that steviol is secreted by renal tubular epithelium, causing diuresis, natriuresis, kaliuresis and a fall in renal tubular reabsorption of glucose. PMID- 19675941 TI - Genetic variation among South Brazilian accessions of Lippia alba Mill. (Verbenaceae) detected by ISSR and RAPD markers. AB - Twenty-seven accessions of Lippia alba Mill. collected in Rio Grande do Sul state, Brazil, were analysed by ISSR and RAPD markers to evaluate their genetic variability and relationships. Six ISSR primers and four RAPD primers generated 120 amplified fragments, most of which were polymorphics. The overall genetic variability among accessions was very high when compared with other plant species. The hierarchical analysis of molecular data (UPGMA) showed low relationship between accessions, and no grouping between accessions of the same chemotype. Canonical functions allowed identifying some variables related with the chemical characteristics of the essential oils. Both ISSR and RAPD markers were efficient to address the genetic diversity of L. alba, and may contribute to the conservation and breeding of this increasingly important aromatic and medicinal species. PMID- 19675942 TI - Daily feeding activity of Bryconamericus microcephalus (Characiformes, Characidae) from Corrego Andorinha, Ilha Grande - RJ. AB - The present study was conducted at the Corrego Andorinha on the eastern slope of Ilha Grande and aimed to determine the alimentary daily rhythm of Bryconamericus microcephalus. Fishes were sampled at a 4 hours interval over a 24 hours period during winter 2002 and summer 2003. Gut fullness was determined according to a numerical scale ranging from 0 (empty gut) to 3 (full gut). No differences were registered for the daily ration estimates between seasons. It is concluded that B. microcephalus is a diurnal feeding species, as the high values of gut fullness occurred between 11:00 AM and 3:00 PM. PMID- 19675943 TI - Effect of histological processing and methacrylate sectioning on the area of gill tissue in teleost. AB - Deformation of biological tissues may occur during histological processing and results in loss of accuracy when quantitative information about cells, tissues and organs is necessary. In this study, the gill tissue from armored catfish (Pterygoplichthys anisitsi) was quantified in each step of processing using the stereological principles. During processing for glycol methacrylate embedding, gill tissue from shrinks significantly but regains its original dimensions after sectioning. PMID- 19675944 TI - Seasonal changes in the somatic indices of the freshwater silverside, Odontesthes bonariensis (Teleostei, Atheriniformes) from a Neotropical shallow lake (Argentina). AB - The study object was an Odontesthes bonariensis (Valenciennes, 1835) population of a characteristically shallow lake from the pampasic region. Over one year, zooplankton and fish samples were collected monthly. As a function of time and gender, several indices were investigated (relative weight, mesentery fat index, gonadosomatic index, hepatosomatic index and somatic index) and were related to zooplanktonic community dynamics, the main energy food source for this species. The results indicate that the seasonal information contained in the weight compartments is most important in mesentery fat, gonads and liver. The zooplankton availability modulates reserves (fat) behaviour and physiologic activity (liver), determining spawn intensity. Thus, in spring spawns, with higher zooplankton availability, mature individuals of the population, males and females, transfer energy from the food to the gonads. In autumn spawns, they use the fat reserves for gonad ripening. PMID- 19675945 TI - Effects of the electromagnetic field, 60 Hz, 3 microT, on the hormonal and metabolic regulation of undernourished pregnant rats. AB - Epidemiological studies have implicated maternal protein-calorie deficiency as an important public health problem in developing countries. Over the last decades, a remarkable diffusion of electricity and an increased level of the electromagnetic field (EMF) in the environment have characterized modern societies. Therefore, researchers are concerned with the biological effects of 50-60 Hz, EMF. The aim of this paper is to show the effects of EMF of 60 Hz, 3 muT, exposure for two hours per day in the regulation of the hormonal and metabolic concentrations in pregnant rats, which were fed by Regional Basic Diet (RBD) during their pregnancy as compared with pregnant rats fed a standard diet. Pregnant rats exposed to EMF of 60 Hz, 3 muT, over the pregnancy and fed with RBD presented an increase in glucose release when compared with the Group subjected only to the RBD ration. Rats fed RBD presented a decrease in their insulin and cortisol serum levels when compared with the Group fed with casein. The T3 and T4 concentrations presented the greatest variation among the Groups. The relation T4:T3 was much exaggerated in the Group subjected to RDB and exposed to EMF when compared to the others. In conclusion, the group subjected to the association of EMF and undernutrition suffered a decrease in its serum concentration of T4 and T3 when compared to the well-nourished group and the relationship T4:T3 in the former group was almost eighteen-fold the later one. PMID- 19675946 TI - The ovary of the bignose fanskate Sympterygia acuta Garman, 1877 (Chondrichthyes, Rajidae) in the Bahia Blanca estuary, Argentina: morphology and reproductive features. AB - Chondrichthyes have become an important economic resource in recent years. In spite of this importance, there is little knowledge about their reproductive biology, especially of species from the south-western Atlantic. In this work, we study the morphology and histology of the ovary of Sympterygia acuta. The results show that oogonia were present only in the immature females (stage I of maturity scale), whereas specimens in stages II and III of maturity displayed oocytes in all developmental steps. The most important histological features that vary throughout oocyte development are the presence or absence of yolk, the number of types and layers of follicular cells and the degree of development of the thecae. Follicular cells are, at least, of two different types. Finally, a new point of view for the determination of maturity stages in the field based on the size of follicles is discussed. PMID- 19675947 TI - Biological indicators of stress in pacu (Piaractus mesopotamicus) after capture. AB - The effects of capture (chasing, netting and air exposure) on cortisol, glucose, chloride, sodium, potassium and calcium concentrations, osmolality, hematocrit, hemoglobin concentration, red blood cells count (RBC) and mean corpuscular volume (MCV) were investigated in pacu (Piaractus mesopotamicus). A total of 132 fish (49.7 +/- 11.7 g) were subjected to capture and 3 minutes air exposure and capture and 5 minutes air exposure. Nine fish at each treatment were sampled at 5, 15, 30, 60 minutes and 24 hours after the procedure. Nine undisturbed fish were sacrificed before the handling and used as controls. Capture resulted in a rise in blood cortisol and glucose 30 and 5 minutes, respectively, after both air exposures. Both indicators returned to resting levels 24 hours after capture. In both fish groups, plasma chloride decreased 60 minutes after capture, not recovering the resting levels within 24 hours after, and serum sodium rose at 15 and 30 minutes and recovered the resting levels 24 hours later. There were no significant changes neither in potassium, calcium and osmolality nor in hematocrit, hemoglobin, RBC and MCV as a consequence of capture. The sequential stressors imposed to pacu during capture activated the brain-pituitary-interrenal axis (cortisol and glucose responses) but the activation of the brain-sympathetic chromaffin cell axis was apparently moderate (ionic and hematological responses). PMID- 19675948 TI - Helminthological records of six-banded armadillos Euphractus sexcinctus (Linnaeus, 1758) from the Brazilian semi-arid region, Patos county, Paraiba state, including new morphological data on Trichohelix tuberculata (Parona and Stossich, 1901) Ortlepp, 1922 and proposal of Hadrostrongylus ransomi nov. comb. AB - This work aimed to evaluate the gastrointestinal helminthfauna composition of six banded armadillos from the Brazilian semi-arid region. Gastrointestinal contents of six road-killed adult animals from Patos County, Paraiba State, were analyzed. Six species of nematodes, comprising five genera and four families, were recovered from the analyzed animals. New morphological data on Trichohelix tuberculata is given, along with a new taxonomical proposal for Hadrostrongylus ransomi (Travassos, 1935) n. comb. This is the first record for parasitic helminths in this host from the Brazilian semi-arid. PMID- 19675949 TI - Seasonal changes in testicular and epididymal histology of the tropical lizard, Tropidurus itambere (Rodrigues, 1987), during its reproductive cycle. AB - The reproductive cycles of lizards, including Tropidurus species, have been widely studied. However, few studies describe in detail the ultrastructure and the epithelial changes in the epididymis. Using histology and trasmission electron microscopy, we show the seasonal changes in the testis and epididymis of the lizard Tropidurus itambere, during its annual reproductive cycle. The reproductive cycle of T. itambere was analysed from June 1988 to June 1989 and from June 2001 to June 2002. While the frequency of reproductive males in the population varied throughout the year, there were reproductive males in most months except for February through April. During this nonreproductive period, there is a reduction in the mean seminiferous tubule volume and few sperm were found in both the testis and the epididymis. PMID- 19675950 TI - Physiological and biochemical aspects of the avian uropygial gland. AB - This review discusses different aspects of the uropygial gland of birds. The gland exhibits a striking morphological diversity in size, shape and presence/absence of tufts of feathers. It was shown that acidic mucins, neutral lipids, glycolipids and phospholipids are normal components of secretion. Several morphological and physiological aspects of the gland were studied on Rock Pigeon Columba livia Gmelin, 1879. The amount of the uropygial gland secretion, its lipid content and fatty acids profile were determined. The extracted lipid mixture contained of C14 to C20 fatty acids, mostly unsaturated; the saturated fatty acids were mainly 14:0, 16:0 and 18:0. No correlation was found between the size of the gland and the aquatic/terrestrial nature of the species. Ablation of the gland did not affect survival, body weight, feeding rate and serum cholesterol, total lipids or calcium levels after 32-120 days. The possible role of the gland in the protection against lipophilic compounds was discussed. The function of the gland is still a subject of controversy. It is accepted that its secretion confers water-repellent properties on the feather coat and maintain the suppleness of the feathers. Other physiological roles of the gland secretion may be associated to pheromone production, control of plumage hygiene, thermal insulation and defence against predators. Concerning the endocrine regulation of the uropygial function, there is scarce information presenting evidence for steroid regulated mechanisms. PMID- 19675951 TI - Genetic diversity and conservation of native populations of Maytenus Ilicifolia Mart. ex Reiss. AB - The aim of this work was to analyze genetic variability in 18 populations of Maytenus ilicifolia, and representatives of Maytenus aquifolia and Maytenus evonymoidis, collected in the states of Mato Grosso do Sul, Parana, Santa Catarina and Rio Grande do Sul, using RAPD molecular markers. Considering total samples of the three species, 263 amplified fragments were identified, of which 72.2% showed to be polymorphous. The index of similarity (Jaccard coefficient) was on average 0.64 between M. ilicifolia and M. aquifolia; 0.47 between M. ilicifolia and M. evonymoidis; and 0.44 between M. aquifolia and M. evonymoidis. The analysis of groupings by the UPGMA algorithm allowed to clearly separate the three analyzed species. In determining the variability in M. ilicifolia, 222 bands were identified, on average 11.1 bands per primer, being 43.2% polymorphous. The index of similarity (Jaccard coefficient) in the bulks of each population in M. ilicifolia was, on average, 0.92 and the index of similarities among the populations was 0.83. The analysis of groupings with the UPGMA algorithm and the analysis of the main coordination (PCO), allowed the separation of the analyzed populations into three groups, the populations from the south of Rio Grande do Sul and the population from Mato Grosso do Sul standing out. A relation between the groupings found and the edaphoclimatic conditions of the collecting places was observed. PMID- 19675952 TI - Rare sexual anomalies in Temora stylifera (Dana, 1849) (Copepoda: Calanoida). PMID- 19675953 TI - Xenomas in Crassostrea rhizophorae (Ostreidae) from Camamu Bay, Bahia, Brazil. PMID- 19675954 TI - Effects of interventions on the demand for hospital services in an influenza pandemic: a sensitivity analysis. AB - PRINCIPLES: The evaluation of the capacity of a country's public health system in case of an influenza pandemic is essential for preparedness planning. Only few studies compare existing medical resources with those required during a severe pandemic. METHODS: We perform a sensitivity analysis with the freely available simulation tool InfluSim to explore the expected number of outpatient visits and the hospital bed occupancy in an influenza pandemic in Switzerland. We define plausible ranges for unknown parameter values and take random samples from these ranges. A set of four simulations is run for each parameter constellation, considering no intervention, contact reduction, antiviral treatment or a combination of both interventions. RESULTS: We find that the peak number of outpatient visits of influenza patients would still be manageable by the current number of active physicians with praxis in Switzerland, and that the demand of hospital beds would be only sustainable in the case of a good compliance of the combined interventions. On the other hand, the demand on intensive care unit beds is unsustainably high. CONCLUSIONS: The range of outcomes, resulting from parameter uncertainty, reaches from outpatient and hospitalization values which are half as high as the median to values which double the median. A combination of antiviral treatment and social distancing can considerably mitigate a severe pandemic, but will only bring it under control for the most optimistic parameter constellation combining (mild outbreaks with a high compliance of interventions). PMID- 19675955 TI - Biofeedback for pain management in traumatised refugees. AB - Chronic pain (CP) and posttraumatic stress disorder (PTSD) are both frequent and often comorbid in refugees. To date, few controlled trials have studied the efficacy of treatments targeting this comorbidity; no treatment guidelines yet exist. The authors examined the feasibility and efficacy of short-term cognitive behavioural biofeedback (BF) addressing CP in traumatised refugees. The sample comprised 11 severely traumatised refugees with CP and PTSD (mean age = 36 years, SD = 6), who underwent assessment with the Mini International Neuropsychiatric Interview, Posttraumatic Diagnostic Scale, Pain Disability Index, and Visual Rating Scale. Additionally, coping with pain and psychotherapy tolerance were assessed. Acceptance of BF was high. Pre-post effects were small to medium for increased pain management and associated heart rate reactivity but large for coping with pain. The results encourage further research to confirm whether BF is indicated as a treatment component, but not a stand-alone treatment, for traumatised refugees with comorbid CP and PTSD. PMID- 19675956 TI - What makes Internet therapy work? AB - Internet therapy is a novel treatment approach that is used to deliver cognitive behaviour therapy. Treatment components are mainly delivered in the form of texts presented via webpages, and support is provided via e-mail. A growing number of controlled trials suggest that Internet therapy works well when (a) a proper diagnosis is made before the treatment starts, (b) a comprehensive treatment is provided, (c) the treatment is user friendly and not overly technically advanced, and (d) support and a clear deadline are provided for the duration of the treatment. Several issues remain for exploration in future research, such as mediating and moderating mechanisms and the role of tailoring the intervention. PMID- 19675957 TI - The mediating role of automatic thoughts in the personality-event-affect relationship. AB - Although cognitive theory gives automatic thoughts a causal role in the onset of negative mood and depressive symptoms, little research has directly tested this relationship, and no research has examined whether automatic thoughts explain the effects of personality factors, life events, and positive mood on negative affect. Accordingly, automatic thoughts were prospectively tested as a mediator of the effects of personality vulnerability factors, positive affect, and hassles on mood. Measures of self-criticism and dependency were administered at baseline, and measures of automatic thoughts, hassles, and positive and negative affect were administered once per week for 4 weeks to 102 college students. Automatic thoughts fully mediated the effects of self-criticism and partially mediated the effects of dependency and hassles on mood. Findings suggest that negative thoughts only partially account for the relationship among personality, hassles, and mood. Results also showed that the impact of positive affect on negative affect may be mediated by the presence or absence of automatic thoughts. PMID- 19675958 TI - Long-term effects of an internet-based treatment for posttraumatic stress. AB - Advances in communication technology offer additional strategies for providing psychological treatment. Previous trials of Internet-based treatment approaches reported significant reductions in posttraumatic stress and related symptoms in response to Internet-based treatments relative to control groups. However, empirical data on the long-term effects of those approaches are sparse. In order to evaluate the long-term effect of an Internet-based intervention, the authors conducted an 18-month follow-up of an Internet-based cognitive behavioural therapy for posttraumatic stress. Severity of posttraumatic stress symptoms was the primary outcome. Additional measures were depression, anxiety, mental and physical health, and health care utilization during the follow-up period. Treatment group participants (n = 34) were assessed 1.5 years after completing treatment. Results indicated that reductions in symptoms of posttraumatic stress symptoms, depression, and anxiety found at posttreatment were sustained during the 18-month follow-up period. Preliminary evidence on long-term effects of Internet-based health care as shown in this study is promising. However, research with larger and clinically more diverse samples is needed to fully assess the clinical impact and potential of Internet-based health care provision. PMID- 19675959 TI - A cluster randomised trial of an internet-based intervention program for tinnitus distress in an industrial setting. AB - The effectiveness of a therapist-supported Internet intervention program for tinnitus distress in an industrial setting was evaluated using a cluster randomised design. Fifty-six Australian employees of two industrial organisations were randomly assigned, based on their work site (18 work sites from BP Australia and five from BHP Billiton), to either a cognitive behavioural therapy (CBT) program or an information-only control program. Participants were assessed at pre and post-program, measuring tinnitus distress, depression, anxiety, stress, quality of life, and occupational health. The CBT program was not found to be superior to the information program for treating tinnitus distress. A high attrition rate and small sample size limit the generalisability of the findings, and further developments of the program and assessment process are needed to enhance engagement and compliance. PMID- 19675960 TI - Cognitive behavioral therapy plus motivational interviewing improves outcome for pediatric obsessive-compulsive disorder: a preliminary study. AB - Lack of motivation may negatively impact cognitive behavioral therapy (CBT) response for pediatric patients with obsessive-compulsive disorder (OCD). Motivational interviewing is a method for interacting with patients in order to decrease their ambivalence and support their self-efficacy in their efforts at behavior change. The authors present a preliminary randomized trial (N = 16) to evaluate the effectiveness of adding motivational interviewing (MI) as an adjunct to CBT. Patients aged 6 to 17 years who were participating in intensive family based CBT for OCD were randomized to receive either CBT plus MI or CBT plus extra psychoeducation (PE) sessions. After four sessions, the mean Children's Yale Brown Obsessive Compulsive Scale (CY-BOCS) score for the CBT plus MI group was significantly lower than for the CBT plus psychoeducation group, t(14) = 2.51, p < .03, Cohen's d = 1.34. In addition, the degree of reduction in CY-BOCS scores was significantly greater, t(14) = 2.14, p = .05, Cohen's d = 1.02, for the CBT plus MI group (mean Delta = 16.75, SD = 9.66) than for the CBT plus psychoeducation group (mean Delta = 8.13, SD = 6.01). This effect decreased over time, and scores at posttreatment were not significantly different. However, participants in the MI group completed treatment on average three sessions earlier than those in the psychoeducation group, providing support for the utility of MI in facilitating rapid improvement and minimizing the burden of treatment for families. PMID- 19675961 TI - Identifying potential barriers to physical activity adherence: anxiety sensitivity and body mass as predictors of fear during exercise. AB - A growing body of work suggests that obese adults are less likely to adhere to exercise than normal-weight adults because they experience greater levels of discomfort and distress during exercise sessions. The present study introduces and provides a preliminary test of the hypothesis that the distress experienced during exercise among persons with elevated body mass index is particularly high among those who fear somatic arousal (i.e. elevated anxiety sensitivity [AS]). Young adults were randomly assigned to complete 20 min of treadmill exercise (at 70% of their age-adjusted predicted maximum heart rate) or 20 min of rest. Body mass, AS, and negative affect were measured at baseline, and fear was measured at 4-min intervals during the experimental phase. Consistent with the authors' hypothesis, there was a significant Exercise x BMI x ASI interaction (sr(2) = .08), suggesting that the greatest fear levels during exercise were observed among participants with high body mass, but only if they also had elevated AS. These findings offer a new approach for identifying specific vulnerable individuals and have clear clinical implications, given that the amplification factor of AS can be modified with clinical intervention. PMID- 19675962 TI - Mediating the effects of cognitive therapy for depression. AB - Cognitive theory holds that inaccurate beliefs and maladaptive information processing play a role in the cause and maintenance of depression, and a cognitive theory of change posits that correcting those errors in thinking will ameliorate existing distress and reduce subsequent risk. Cognitive therapy has been shown to be efficacious in the treatment of depression and prevention of subsequent relapse, but evidence for mediation has been difficult to detect. The authors review efforts to test for mediation in cognitive therapy and describe the epistemological issues that complicate the process. Cognitive mediation of acute response likely will be hard to detect, whereas there already is good evidence that cognitive processes play a role in the mediation of enduring effects. PMID- 19675963 TI - Ambivalence, silence and gender differences in church leaders' HIV-prevention messages to young people in KwaZulu-Natal, South Africa. AB - A series of semi-structured interviews on HIV prevention were conducted with South African clergy with pastoral and liturgical responsibilities from the Roman Catholic Church, the Lutheran Church and the Assemblies of God. The interviews were tape-recorded, transcribed verbatim and analysed by interpretive descriptive analysis. Three themes indicative of church leaders' approach to HIV prevention among youth emerged: dilemmas in breaking the silence on HIV and AIDS; ambivalent HIV-prevention messages from church leaders to young people; and gender differences in HIV-prevention messages. While church leaders had taken steps to overcome the stigma, the dilemmas of balancing theological understanding with resistance from their congregations presented a complex scenario. Ambivalence to HIV prevention concerned whose responsibility it was to educate young people about HIV; talking about sexuality in public; pre-marital abstinence and condom use; and resistance from congregation members towards HIV prevention. Finally, findings indicated a discrepancy between church leaders' belief in gender equality and the HIV-prevention messages they verbalised, which appears to burden girls. PMID- 19675967 TI - Chrysosporium guarroi sp. nov. a new emerging pathogen of pet green iguanas (Iguana iguana). AB - Chrysosporium guarroi sp. nov. represented by five strains isolated from cases of dermatomycosis in pet green iguanas (Iguana iguana) in Spain, is described and illustrated. This taxon is characterized by its ability to grow at temperatures from 15 to 37 degrees C and by the presence of arthroconidia and aleurioconidia. The latter are unicellular, smooth, pyriform or clavate, sessile or borne at the ends of narrow stalks. The analysis of the sequences of the D1/D2 and ITS regions confirm the separation of this new species from others of the genus Chrysosporium. PMID- 19675968 TI - The prevalence and quality of silent, socially silent, and disclosed autobiographical memories across adulthood. AB - Two separate studies examined the prevalence and quality of silent (infrequently recalled), socially silent (i.e., recalled but not shared), and disclosed autobiographical memories. In Study 1 young and older men and women remembered positive events. Positive memories were more likely to be disclosed than to be kept socially silent or completely silent. However, socially silent and disclosed memories did not differ in memory quality: the memories were equally vivid, significant, and emotional. Silent memories were less qualitatively rich. This pattern of results was generally replicated in Study 2 with a lifespan sample for both positive and negative memories, and with additional qualitative variables. The exception was that negative memories were kept silent more often. Age differences were minimal. Women disclosed their autobiographical memories more, but men told a greater variety of people. Results are discussed in terms of the functions that memory telling and silences might serve for individuals. PMID- 19675969 TI - Characteristics and reference ranges of Insulin-Like Growth Factor-I measured with a commercially available immunoassay in 724 healthy adult Caucasians. AB - BACKGROUND AND OBJECTIVE: Measurements of Insulin-Like Growth Factor-I (IGF-I) play a pivotal role in the evaluation of the growth hormone-IGF-I axis. Due to assay variation IGF-I reference ranges are assay specific. We provide serum IGF-I reference ranges for adult men and women obtained by a commercially available assay. METHOD: IGF-I was measured by an enzyme-linked immunosorbent assay (R&D Systems). Assay precision was evaluated in low, medium and high IGF-I pools and in single samples from outpatients. The reference ranges were obtained in 724 healthy Caucasians, mean age 48 years (range 19-91). RESULTS: IGF-I was measured higher in serum compared to plasma samples (p<0.001, R2=0.96). To convert EDTA plasma values to serum values a factor of 1.22 was calculated. The intraassay coefficients of variation (CV) for the low: 56.4+/-2.7 ng/mL, medium: 179.7+/-5.9 ng/mL and high pool: 445.61+/-3.2 ng/mL (mean 1 SD) were 5, 3 and 3%. Interassays CVs for the low, medium and high pool varied between 7-10, 5-7, and 6-9%. Reproducibility between 4 different lots showed a intraclass CV of 0.99 (95%CI 0.98-0.96). Logarithmically transformed IGF-I levels were linearly associated with age with a 13% reduction in IGF-I per decade in females and 11% in males. CONCLUSION: We have provided IGF-I reference ranges obtained by an assay that showed variations and reproducibility that was considered of sufficient high quality for clinical and scientific use. PMID- 19675971 TI - The effects of the second generation antipsychotics and a typical neuroleptic on collagen-induced platelet aggregation in vitro. AB - OBJECTIVE: Antipsychotics are widely used in psychiatry, and consequently a lot of their side effects have been reported. One of them is cardiovascular disease leading to increased risk of stroke, thrombosis, pulmonary, embolism, in which hyperactivation of blood platelets is involved. The purpose of the present study was to examine the effects of the second generation antipsychotics (SGAs) such as clozapine, risperidone, and olanzapine, and a typical neuroleptic - haloperidol - on the one step of platelet activation-platelet aggregation induced by collagen in vitro. Blood was collected into buffered sodium citrate (3.8%) and centrifuged to get platelet-rich plasma (PRP). In PRP (2x10(8) platelets/ml) obtained from healthy volunteers that was incubated with antipsychotics (clozapine, risperidone, olanzapine, haloperidol; 30 min) aggregation of blood platelets was measured using a Chrono-Log Lumi-aggregometer. Aggregation of platelets was measured after stimulation of platelets with 1 microl of collagen (2 microg/ml). RESULTS: Clozapine, like haloperidol reduced platelet aggregation induced by collagen (inhibition of platelet aggregation reached about 20%) (P=1x10(-5) and P=0.003, respectively). Risperidone had also a weak antiaggregatory effect (P=0.05). Among tested antipsychotics only olanzapine had no effect on collagen stimulated platelet aggregation (P>0.05). CONCLUSION: The obtained results indicate that the difference in action of tested drugs on platelet aggregation may dependent on the various chemical structures of these drugs. Clozapine, risperidone and haloperidol are structurally diverse, and they all significantly reduce platelet aggregability induced by collagen. On the other hand, a close structural analog of clozapine - olanzapine - did not inhibit platelet aggregation. However, mechanism of antipsychotics action on blood platelets is not clear. Moreover, it seems that clozapine, risperidone and haloperidol treatment due to antiaggregatory action may have even some antithrombotic effects. PMID- 19675973 TI - Three trends that undermine the Greek health system: is there a way out? PMID- 19675974 TI - Faculty development for teachers of family medicine in Europe: reflections on 16 years' experience with the international Bled course. AB - BACKGROUND: Instruction in principles and methods of medical education is a core component of the training of medical teachers. Curricula for new medical teachers have developed across Europe, but few European courses have had the success of the EURACT Bled course for teachers of family medicine. The course focuses on practical issues in medicine and the professional development of physicians. This article describes 16 years' experience with the Bled course for teachers in general practice/family medicine (GP/FM). COURSE STRUCTURE: The course is centred on the preparation of a teaching module, exploring a specific theme in family medicine. The main teaching methods used are: keynote lectures, small group sessions, field work and preparation of a teaching module by the participants. OUTCOMES: This course has attracted 555 participants from 20 countries since 1992. The course and its outputs have been applied in undergraduate and postgraduate teaching, and in continuous professional development (CPD) in several countries. It is a respected forum for faculty development. The experience of the Bled course suggests that academic medicine may be better able to fulfil its responsibilities by paying attention to relevant topics and using appropriate methods in undergraduate and postgraduate medical curricula. PMID- 19675975 TI - The preferences and rationale of family doctors in pharmacological treatment for depression. PMID- 19675976 TI - Computational fluid dynamics of aggregating red blood cells in postcapillary venules. AB - Aggregate formation of red blood cells (RBCs) in a postcapillary venular bifurcation is investigated with three-dimensional computer simulations using the Chimera grid method. Interaction energy between the RBCs is modelled by a depletion interaction theory; RBCs are modelled as rigid oblate ellipsoids. The cell-cell interactions of RBCs are strongly dependent on vessel geometry and shear rates. The experimental data on vessel geometry, pseudoshear rates, and Dextran concentration obtained in our previous in vivo RBC aggregation study in postcapillary venules of the rat spinotrapezius muscle were used to simulate RBC aggregation. The computational results were compared to the experimental results from the in vivo study. The results show that cells have a larger tendency to form an aggregate under reduced flows. Aggregate formation also depends on the angle and location of the cells before they enter the bifurcation region. Comparisons with experimental data are discussed. PMID- 19675977 TI - Does placenta position modify the risk of placental abruption in car crashes? AB - The goal of this paper is to study the influence of placenta position on the risk of placenta abruption following a car crash involving a full term pregnant woman in the driver's seat. We developed an anatomically precise finite element numerical model of a pregnant woman. This numerical model was validated using experimental data from crash tests using cadavers given to science. For a frontal impact at a speed of 20 km/h, the mean value of peak strain levels at the utero placenta interface were found to be close to 20%, independent of placenta position, representing an adverse foetal outcome risk of approximately 15%. This study found no significant effect of placenta position on the risk of placenta abruption. Our anatomically precise approach does however confirm the interest of using a numerical model when studying injury mechanisms in pregnant women involved in car crashes. PMID- 19675979 TI - Characterisation of a phenomenological model for commercial pneumatic muscle actuators. AB - This study focuses on the parameter characterisation of a three-element phenomenological model for commercially available pneumatic muscle actuators (PMAs). This model consists of a spring, damping and contractile element arranged in parallel. Data collected from static loading, contraction and relaxation experiments were fitted to theoretical solutions of the governing equation for the three-element model resulting in prediction profiles for the spring, damping and contractile force coefficient. For the spring coefficient, K N/mm, the following relationships were found: K = 32.7 - 0.0321P for 150 < or = P < or = 314 kPa and K = 17 + 0.0179P for 314 < or = P < or = 550 kPa. For the damping coefficient, B Ns/mm, the following relationship was found during contraction: B = 2.90 for 150 < or = P < or = 550 kPa. During relaxation, B = 1.57 for 150 < or = P < or = 372 kPa and B = 0.311 + 0.00338P for 372 < or = P < or = 550. The following relationship for the contractile force coefficient, F(ce) N, was also determined: F(ce) = 2.91P+44.6 for 150 < or = P < or = 550 kPa. The model was then validated by reasonably predicting the response of the PMA to a triangular wave input in pressure under a constant load on a dynamic test station. PMID- 19675978 TI - New resource for the computation of cartilage biphasic material properties with the interpolant response surface method. AB - Cartilage material properties are important for understanding joint function and diseases, but can be challenging to obtain. Three biphasic material properties (aggregate modulus, Poisson's ratio and permeability) can be determined using an analytical or finite element model combined with optimisation to find the material properties values that best reproduce an experimental creep curve. The purpose of this study was to develop an easy-to-use resource to determine biphasic cartilage material properties. A Cartilage Interpolant Response Surface was generated from interpolation of finite element simulations of creep indentation tests. Creep indentation tests were performed on five sites across a tibial plateau. A least-squares residual search of the Cartilage Interpolant Response Surface resulted in a best-fit curve for each experimental condition with corresponding material properties. These sites provided a representative range of aggregate moduli (0.48-1.58 MPa), Poisson's ratio (0.00-0.05) and permeability (1.7 x 10(- 15)-5.4 x 10(- 15) m(4)/N s) values found in human cartilage. The resource is freely available from https://simtk.org/home/va squish. PMID- 19675981 TI - New directions in research bring new clinical challenges. PMID- 19675980 TI - Influence of inlet boundary conditions on the local haemodynamics of intracranial aneurysms. AB - Haemodynamics is believed to play an important role in the initiation, growth and rupture of intracranial aneurysms. In this context, computational haemodynamics has been extensively used in an effort to establish correlations between flow variables and clinical outcome. It is common practice in the application of Dirichlet boundary conditions at domain inlets to specify transient velocities as either a flat (plug) profile or a spatially developed profile based on Womersley's analytical solution. This paper provides comparative haemodynamics measures for three typical cerebral aneurysms. Three dimentional rotational angiography images of aneurysms at three common locations, viz. basilar artery tip, internal carotid artery and middle cerebral artery were obtained. The computational tools being developed in the European project @neurIST were used to reconstruct the fluid domains and solve the unsteady Navier-Stokes equations, using in turn Womersley and plug-flow inlet velocity profiles. The effects of these assumptions were analysed and compared in terms of relevant haemodynamic variables within the aneurismal sac. For the aneurysm at the basilar tip geometries with different extensions of the afferent vasculature were considered to study the plausibility of a fully-developed axial flow at the inlet boundaries. The study shows that assumptions made on the velocity profile while specifying inlet boundary conditions have little influence on the local haemodynamics in the aneurysm, provided that a sufficient extension of the afferent vasculature is considered and that geometry is the primary determinant of the flow field within the aneurismal sac. For real geometries the Womersley profile is at best an unnecessary over-complication, and may even be worse than the plug profile in some anatomical locations (e.g. basilar confluence). PMID- 19675982 TI - Casting a vision for amputation rehabilitation. PMID- 19675983 TI - Lower-limb amputee needs assessment using multistakeholder focus-group approach. AB - To assess the needs of lower-limb amputees and identify differences between diabetic dysvascular amputees and traumatic amputees, we held a multistakeholder focus-group workshop whose participants included veteran lower-limb amputees, clinicians, researchers, and prosthetic device manufacturers. We conducted the initial workshop sessions as traditional focus-group meetings with homogeneous participant groups generating lists of issues relevant to the individual groups. Subsequent sessions assembled heterogeneous participant groups for a two-phase approach: Discovery and Codesign. The Discovery phase used observation and discussion to elicit specific needs. The Codesign phase focused on emergent topics and explored potential solutions. The participants identified needs associated with desired improvements to the socket system, foot and ankle components, and alignment with the residual limb. One need was a comprehensive understanding of the recovery path following amputation that could be addressed through enhanced education and communication. Another need was remote monitoring systems that could potentially improve quality of care. No dichotomy of needs between diabetic dysvascular amputees and traumatic amputees was evident among the participants of this workshop. The lively, open-ended discussions produced numerous suggestions for improving amputee quality of life that are listed to facilitate future research and development. PMID- 19675984 TI - Assessing the state of clinically applicable research for evidence-based practice in prosthetics and orthotics. AB - Evidence-based practice combines a practitioner's training and experience with evidence established through scientific research. Fundamental to the evidence based process for prosthetics and orthotics is the ongoing availability of clinically applicable research on relevant conditions, components, and patient populations. In the past, research has been successfully applied to practice, sometimes producing substantial changes. Examples include clinically applicable research that has assessed treatment effectiveness, altered clinical patient interaction, led to the development of new components and technologies, and challenged or changed long-standing clinical opinion. Despite past successes, obstacles remain in the application of research to practice. Practitioners have stated a desire for research and have identified a list of research needs but lack the training or resources necessary to conduct the research. A gulf also exists between the perceived research needs and the clinically applicable research that is being produced, possibly because of the broad nature of those needs. PMID- 19675985 TI - Mathematical modeling and mechanical and histopathological testing of porous prosthetic pylon for direct skeletal attachment. AB - This article presents recent results in the development of the skin and bone integrated pylon (SBIP) intended for direct skeletal attachment of limb prostheses. In our previous studies of the porous SBIP-1 and SBIP-2 prototypes, the bond site between the porous pylons and residuum bone and skin did not show the inflammation characteristically observed when solid pylons are used. At the same time, porosity diminished the strength of the pylon. To find a reasonable balance between the biological conductivity and the strength of the porous pylon, we developed a mathematical model of the composite permeable structure. A novel manufacturing process was implemented, and the new SBIP-3 prototype was tested mechanically. The minimal strength requirements established earlier for the SBIP were exceeded threefold. The first histopathological analysis of skin, bone, and the implanted SBIP-2 pylons was conducted on two rats and one cat. The histopathological analysis provided new evidence of inflammation-free, deep ingrowth of skin and bone cells throughout the SBIP structure. PMID- 19675986 TI - One hundred patients treated with osseointegrated transfemoral amputation prostheses--rehabilitation perspective. AB - Treatment with osseointegrated transfemoral prostheses has been shown to improve quality of life. The treatment has been performed in Sweden since 1990 and consists of two surgical procedures followed by rehabilitation. During the first years, the rehabilitation process was not standardized. In 1999, a treatment protocol called OPRA (Osseointegrated Prostheses for the Rehabilitation of Amputees) was established. This article describes the current rehabilitation protocol and illustrates the overall results. The OPRA rehabilitation protocol is graded to stimulate the process of osseointegration and prepare the patient for unrestricted prosthetic use. It includes initial training with a short training prosthesis followed by gradually increased prosthetic activity. Between May 1990 and June 2008, we treated 100 patients with 106 implants (6 bilaterally; 61% males, 39% females; mean age 43 years; mean time since amputation 11.5 years.) The majority had amputations due to trauma (67%) or tumor (21%) (other = 12%). Currently, 68 patients are using their prostheses (follow-up: 3 months- 17.5 years) and 32 are not (4 are deceased, 7 are before second surgery, 6 are in initial training, 4 are not using prosthesis, and 11 had the implant removed). The majority of treatment failures occurred in patients before we established the OPRA protocol. The implementation of graded rehabilitation is considered to be of utmost importance for improved results. PMID- 19675988 TI - Agonist-antagonist active knee prosthesis: a preliminary study in level-ground walking. AB - We present a powered knee prosthesis with two series-elastic actuators positioned in parallel in an agonist-antagonist arrangement. To motivate the knee's design, we developed a prosthetic knee model that comprises a variable damper and two series-elastic clutch units that span the knee joint. Using human gait data to constrain the model's joint to move biologically, we varied model parameters using an optimization scheme that minimized the sum over time of the squared difference between the model's joint torque and biological knee values. We then used these optimized values to specify the mechanical and control design of the prosthesis for level-ground walking. We hypothesized that a variable-impedance control design could produce humanlike knee mechanics during steady-state level ground walking. As a preliminary evaluation of this hypothesis, we compared the prosthetic knee mechanics of an amputee walking at a self-selected gait speed with those of a weight- and height-matched nonamputee. We found qualitative agreement between prosthetic and human knee mechanics. Because the knee's motors never perform positive work on the knee joint throughout the level-ground gait cycle, the knee's electrical power requirement is modest in walking (8 W), decreasing the size of the onboard battery required to power the prosthesis. PMID- 19675989 TI - Transtibial amputee joint rotation moments during straight-line walking and a common turning task with and without a torsion adapter. AB - Amputees lack movement and control mechanisms at the foot and ankle that result in different strategies for locomotion than nonamputees. The torsion adapter is a prosthetic device designed to minimize shear stress at the residual limb by facilitating rotation in the transverse plane. This study determined if the addition of a torsion adapter alters lower-limb joint rotation moments of transtibial amputees walking in a straight line and turning. Ten transtibial amputees wore either a torsion adapter or a rigid adapter for an acclimation period of 3 weeks in random order. Ten nonamputees were also included for comparison. Kinetics were collected as participants walked in a straight line and around a 1 m-radius circular path at their self-selected turning walking speed. When amputee participants wore the torsion adapter, they demonstrated decreased prosthetic-limb peak internal rotation moments at the inside limb knee and hip compared with when they wore the rigid adapter, which may facilitate changes in orientation by not actively resisting the turn. Nonamputees exhibited larger moments compared with the prosthetic limb for both the amputee participants wearing either the torsion or rigid adapters. No differences were found in the moments for the intact limb between torsion and rigid adapter conditions during turning and for both limbs during straight-line walking. PMID- 19675990 TI - Early delayed amputation: a paradigm shift in the limb-salvage time line for patients with major upper-limb injury. AB - Patients with major injuries to the upper limbs sometimes fail to achieve successful limb salvage. During the attempt to fashion a functional limb, multiple painful procedures may be ventured. Despite the best efforts of surgeons and therapists, a nonfunctioning or painful upper limb may remain in place for many months or years before late delayed amputation and progression to productive rehabilitation occur. We present three patient cases that illustrate failed upper limb salvage. In each case, patients expressed a desire for amputation at 6 months after their injury. To reduce the pain and suffering that patients with failed limb salvage endure, we propose a paradigm shift in the limb-salvage time line. We suggest that patients be evaluated for early delayed amputation 6 months after their injury. PMID- 19675987 TI - On the way to total integration of prosthetic pylon with residuum. AB - Two decades after introducing threaded titanium dental implants, Dr. Per-Ingvar Branemark used a similar technique in the 1980s to pioneer the direct skeletal attachment (DSA) of limb prostheses. He and his colleagues used convincing clinical experience to overcome the skepticism of their peers, affording a new dimension of prosthetic rehabilitation to almost 100 individuals with amputation. As a result, more research has been initiated worldwide to move DSA to a level of greater safety, longevity, and reliability. This review highlights the trends and milestones in current DSA development. It also identifies ideas from previous studies in various fields that may be useful in future DSA development. PMID- 19675991 TI - A new option for amputees: transplantation of the hand. AB - The permanent loss of a hand or limb results in significant challenges. A number of options are available to individuals who must deal with this loss, including reconstructive surgery using a person's own tissue to repair the damage or the use of prosthetic devices. We present an update on the most recent addition to the list of options, namely, composite tissue allotransplantation (CTA). In this procedure, tissue to repair the loss is taken from deceased donors who are giving hearts, kidneys, and tissue for transplantation. We report on the world's longest follow-up of CTA of the hand, as well as four other American hand transplant recipients. In very select patient populations, we propose that transplantation is now a clinical option for amputees. PMID- 19675992 TI - Dynamic interface pressure distributions of two transtibial prosthetic socket concepts. AB - In this study, we investigated and compared the dynamic interface pressure distribution of hands-off and hands-on transtibial prosthetic systems by means of pressure mapping. Of the 48 established unilateral amputees recruited, half (n = 24) had been wearing pressure-cast prostheses (IceCast Compact) and the other half (n = 24) had been wearing hand-cast sockets of the patellar tendon bearing design. We measured the dynamic pressure profile of more than 90% of the area within each prosthetic socket by means of four Tekscan F-Scan socket transducer arrays. We compared the interface pressure between socket concepts. We found that the distribution of dynamic pressure at the limb-socket interface was similar for the two intervention (socket prescription) groups. However, a significant difference was found in the magnitude of the interface pressure between the two socket concepts; the interface pressures recorded in the hands-off sockets were higher than those seen in the hands-on concept. Despite the differences in interface pressure, the level of satisfaction with the sockets was similar between subject groups. The sockets instrumented for this study had been in daily use for at least 6 months, with no residual-limb health problems. PMID- 19675993 TI - Differences in function and safety between Medicare Functional Classification Level-2 and -3 transfemoral amputees and influence of prosthetic knee joint control. AB - The functional differences between persons with amputation who are classified as Medicare Functional Classification Level (MFCL)-2 and -3 include the abilities to walk at various cadences and to negotiate environmental barriers outside the home. This study compared the effect of active microprocessor control and passive mechanical control of the prosthetic knee on function and safety in 17 subjects with transfemoral amputation (8 MFCL-2 and 9 MFCL-3). Assessed functional tasks included hill and stair descent, an attentional demand task, and an obstacle course. Self-reported measures included concentration, multitasking ability, and numbers of stumbles and falls. Active knee control was associated with significant improvements (p < 0.05) in hill and stair gait, speed (hills, obstacle course, and attentional demand task), and ability to multitask while walking for both cohorts. MFCL-2 subjects also reported a significant reduction (p < 0.01) in uncontrolled falls. Over the study, 50% of MFCL-2 subjects and 33% of MFCL-3 subjects transitioned to a higher MFCL. Results suggest that active knee control improves function and reduces the frequency of adverse events in a population that is at risk for falls. Use of active knee control may allow persons with amputation to expand their functional domain, transition to a higher MFCL, and access additional prosthetic options. PMID- 19675994 TI - Walking mechanics of persons who use reciprocating gait orthoses. AB - Although ambulation with a reciprocating gait orthosis (RGO) may provide physical benefits to people with lower-limb paralysis, the high metabolic energy cost associated with ambulation limits orthosis use. The purpose of this case series was to investigate the dynamics of ambulation with RGOs to identify and better understand the potential causes of the high energy cost. Data were acquired from five regular users of RGOs. Kinematics and kinetics were measured, and the moments and powers acting at the hips and shoulders calculated. All RGO users walked with a flexed trunk and bore a large proportion of body weight through the arms during single support. Moments at the shoulder encouraged trunk extension, while moments at the hip encouraged trunk flexion. An extension moment acted on the hip at the beginning of swing, which was antagonistic to the goal of swing and contradicted the intent of the reciprocal link: to advance the swing leg. These results suggest that characteristics of RGO ambulation are consistent across users. The relationship between posture, forces acting on the walking aids, and the action of the RGO reciprocal link should be further explored because these factors likely contribute to the high metabolic cost of ambulation with an RGO. PMID- 19675995 TI - Development of hybrid orthosis for standing, walking, and stair climbing after spinal cord injury. AB - This study explores the feasibility of a hybrid system of exoskeletal bracing and multichannel functional electrical stimulation (FES) to facilitate standing, walking, and stair climbing after spinal cord injury (SCI). The orthotic components consist of electromechanical joints that lock and unlock automatically to provide upright stability and free movement powered by FES. Preliminary results from a prototype device on nondisabled and SCI volunteers are presented. A novel variable coupling hip-reciprocating mechanism either acts as a standard reciprocating gait orthosis or allows each hip to independently lock or rotate freely. Rotary actuators at each hip are configured in a closed hydraulic circuit and regulated by a finite state postural controller based on real-time sensor information. The knee mechanism locks during stance to prevent collapse and unlocks during swing, while the ankle is constrained to move in the sagittal plane under FES-only control. The trunk is fixed in a rigid corset, and new ankle and trunk mechanisms are under development. Because the exoskeletal control mechanisms were built from off-the-shelf components, weight and cosmesis specifications for clinical use have not been met, although the power requirements are low enough to provide more than 4 hours of continuous operation with standard camcorder batteries. PMID- 19675996 TI - Autoimmune hepatitis. PMID- 19675997 TI - Autoantibodies and their antigens in autoimmune hepatitis. AB - Autoantibody detection assists in the diagnosis and allows differentiation of autoimmune hepatitis (AIH) type 1 (AIH-1), characterized by antinuclear antibody (ANA) and/or smooth muscle antibody (SMA), and type 2 (AIH-2), distinguished by the presence of antibodies to liver-kidney microsome type 1 (anti-LKM1) and/or antibodies to liver cytosol type 1 (anti-LC1). Detection of atypical perinuclear antineutrophil cytoplasmic antibodies (pANCA) and anti-soluble liver antigen (SLA) antibodies can act as an additional pointer toward the diagnosis of AIH, particularly in the absence of the conventional autoantibodies. Routine autoantibody testing by indirect immunofluorescence has been recently complemented by molecular assays based on purified or recombinant antigens. Although the AIH-1-specific ANA and SMA targets need better definition, those of anti-LKM1 and anti-LC1 in AIH-2 have been clearly identified; the fine specificity of antibody reactivity and its clinical relevance to disease pathogenesis are the focus of ongoing investigation. This article critically discusses the current knowledge of the diagnostic and clinical significance of AIH-related autoantibody reactivities, focusing on key issues that the physician needs to be aware of to be able to request the appropriate testing and to interpret correctly the laboratory results within the clinical context of the patient. PMID- 19675998 TI - Scoring systems for the diagnosis of autoimmune hepatitis: past, present, and future. AB - The diagnosis of autoimmune hepatitis (AIH) can be challenging due to the variable clinical and laboratory findings. The original diagnostic criteria published in 1993 by the International Autoimmune Hepatitis Group (IAIHG) were revised in 1999 in an attempt to standardize the diagnosis. However, these criteria are complex and can be cumbersome in clinical practice. In 2008, simplified diagnostic criteria were reported to facilitate the bedside diagnosis of AIH. The scoring systems have been evaluated in several retrospective case series and tested for their ability to reliably diagnose and exclude AIH. However, the scoring systems did not fare as well in patients with concomitant cholestatic disease, fatty liver disease, fulminant hepatitis, and pediatric patients. Both positive and negative predictive values are low in these patients. Prospective studies are needed to compare the two scoring systems to determine which (if not all) patients require liver biopsy and which patients would benefit from immunotherapy. PMID- 19675999 TI - New animal models for autoimmune hepatitis. AB - Autoimmune hepatitis (AIH) is often diagnosed late in the disease course and usually requires lifelong immunosuppressive therapy. Unfortunately, the etiology of the disease and the mechanisms leading to the autoimmune destruction of the liver parenchyma are only poorly understood. For a long time, one reason for this lack of apprehension was the absence of reliable animal models with a chronic immune response against liver tissues. Initial attempts to break tolerance against hepatocytes usually just resulted in mild, transient hepatitis flares. Recently, however, some approaches have been made to establish models of chronic AIH that reflect the immunopathogenic mechanisms seen in humans. In this article, we reflect on recent models, focusing on their feasibility and chances for success in providing a platform for studying the mechanisms of autoimmune liver destruction and the development of possible therapeutic interventions. PMID- 19676000 TI - Treatment of autoimmune hepatitis. AB - First described in 1950, autoimmune hepatitis (AIH) was the first chronic liver disease with a favorable response to drug therapy and a dismal prognosis when left untreated. In the decades since the first treatment studies, the basic strategic principle of inducing remission with steroids and azathioprine has not been modified. The main problems are a timely diagnosis before cirrhosis develops, the avoidance of immunosuppressant side effects, nonresponders to standard induction therapy, and adherence to therapy. Alternative drugs for immunosuppression have been tested in small series and include transplant immunosuppressants. A large multicenter prospective treatment trial suggests that budesonide may offer an alternative in noncirrhotic AIH patients and can minimize unwanted steroid effects. The ultimate treatment approach upon drug treatment failure is liver transplantation. Although only 4% of transplant candidates are AIH patients, the risk for graft loss because of recurrence has to be considered, and recurrent AIH should be treated after transplantation. PMID- 19676001 TI - Autoimmune hepatitis and liver transplantation: indications, results, and management of recurrent disease. AB - For those with autoimmune hepatitis (AIH), indications for liver transplantation include end-stage liver failure (as suggested by a MELD score >16), the onset of liver cancer, intractable symptoms that make the patient's life intolerable, and fulminant liver failure; outcomes are excellent, with 10-year survival in excess of 70%. For those with a fulminant presentation, the impact of corticosteroids is controversial and liver transplantation may be required. Autoimmune hepatitis recurs in approximately one third and may be detected up to 10 years or more post transplant. There are no agreed-on criteria for the diagnosis, and differentiation from rejection and other causes of graft damage can be difficult. There is no strong correlation between the prevalence of recurrent AIH (rAIH) and graft loss from rAIH. Treatment of recurrent disease with the addition or increase in corticosteroids is often successful, although long-term data are lacking and some may progress to graft failure despite increased treatment. There remains controversy over the role for protocol liver biopsies to detect recurrent disease and the best immunosuppressive strategies to prevent and treat recurrence. PMID- 19676002 TI - Autoimmune hepatitis in children: what is different from adult AIH? AB - Autoimmune hepatitis (AIH) is characterized by inflammatory liver histology, circulating non-organ-specific autoantibodies, and increased levels of immunoglobulin (Ig) G in the absence of a known etiology. Two types of childhood AIH are recognized according to seropositivity: smooth muscle antibody (SMA) and/or antinuclear antibody (ANA), which is AIH type 1; and antibodies to liver kidney microsome type 1 (anti-LKM1), which is AIH type 2. There is a female predominance in both. Autoimmune hepatitis type 2 presents more acutely, at a younger age, and commonly with IgA deficiency; however, duration of symptoms before diagnosis, clinical signs, family history of autoimmunity, presence of associated autoimmune disorders, response to treatment, and long-term prognosis are similar in the two groups. Immunosuppressive treatment with steroids and azathioprine, which should be instituted promptly to avoid progression to cirrhosis, induces remission in 80% of cases. Relapses are common, often due to nonadherence. Drugs effective in refractory cases include cyclosporine and mycophenolate mofetil. Long-term treatment is usually required, with only some 20% of AIH type 1 patients able to discontinue therapy successfully. In childhood, sclerosing cholangitis with strong autoimmune features, including interface hepatitis and serological features identical to AIH type 1, is as prevalent as AIH, but it affects boys and girls equally. The differential diagnosis relies on cholangiographic studies. In autoimmune sclerosing cholangitis, liver parenchymal damage responds satisfactorily to immunosuppressive treatment, whereas bile duct disease tends to progress. PMID- 19676003 TI - The autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy or autoimmune polyglandular syndrome type 1. AB - Autoimmune polyglandular syndromes are rare autoimmune endocrinopathies that are associated with nonendocrine autoimmunopathies. Autoimmune polyendocrinopathy candidiasis-ectodermal dystrophy (APECED), also named autoimmune polyglandular syndrome type 1 (APS-1), is distinguished from autoimmune polyglandular syndrome 2 (APS-2). Major disease components of APECED are adrenal insufficiency, hypoparathyroidism, and candidiasis. The diagnosis is established by the presence of two out of the three components. Minor clinical features include autoimmune hepatitis, which occurs in up to 20% of APECED patients, and ranges from a mild to a fulminant course. The disease mostly affects juvenile patients from Sardegna, Italy, Finland, and Iran (Iranian Jews), but it also occurs in other ethnic groups. The AIRE gene responsible for APECED is expressed in cells involved in induction and maintenance of immune tolerance. Genetic alterations of the single gene are associated with APECED. Because a specific therapy is not currently available, treatment consists of hormone replacement and caring for clinical symptoms. PMID- 19676004 TI - Special clinical challenges in autoimmune hepatitis: the elderly, males, pregnancy, mild disease, fulminant onset, and nonwhite patients. AB - Autoimmune hepatitis has diverse presentations that may mask its identity, affect its clinical behavior, and confound its management. The objectives of this report are to characterize the populations that are underdiagnosed or difficult to manage and to provide guidelines for meeting their challenges. The elderly have an aggressive, indolent liver disease that responds well to conventional corticosteroid treatment, but the diagnosis of this disease can be hampered by concurrent illnesses and medication. Genetic factors affect the expression and outcome of autoimmune hepatitis in different genders, and they may influence the nature and reactivity to triggering antigens or hormonal imbalances. Pregnancy can ameliorate autoimmune hepatitis, whereas delivery can exacerbate it. Asymptomatic patients and those with mild laboratory derangements may not require immediate therapy, but they commonly require treatment later. Centrilobular zone 3 necrosis can characterize an acute, severe, or fulminant presentation, and corticosteroid therapy can be lifesaving if effective within 2 weeks. Region specific etiological agents, host-dependent susceptibility factors, and socioeconomic conditions may diversify the presentation in different ethnic groups. The special clinical challenges are essential to recognize and treat properly, and they may also extend hypotheses of pathogenesis. PMID- 19676005 TI - Vaccination as a triggering event for autoimmune hepatitis. AB - The pathogenesis of autoimmune hepatitis entails complex interactions between triggering factors, autoantigens, genetic predisposition, and immunoregulatory networks. Implicated triggering factors are numerous and include toxins, medications, and infectious agents. In this article, we present a unique case of a 31-year-old woman with severe autoimmune hepatitis apparently abruptly triggered by vaccination. All evidence suggests this was an idiosyncratic response to either hepatitis A or yellow fever vaccination. PMID- 19676006 TI - [Early recognition and prevention of psychoses]. PMID- 19676007 TI - [Efficiency of outpatient and inpatient alcohol treatment - predictors of outcome]. AB - AIM: To study the prediction potential of demographic and psychopathological variables in the patient allocation context. METHOD: This was a follow-up study of alcohol-dependent patients (n = 242) who were treated in an outpatient (n = 75), inpatient short-term (n = 88), or inpatient long-term (n = 79) setting between 2003 and 2004. Demographic and psychopathological variables were measured by psychiatric basic documentation and the European Addiction Severity Index (EuropASI) . Abstinence two years after discharge from treatment was the primary outcome criterion. RESULTS: Significant predictors of outcome were a higher secondary school qualification, a finished professional training, later onset of alcohol dependence, less number of treatments for physical illness as well as no history of suicide attempts. Both satisfaction with the family background and being employed were predictors with allocation potential. PMID- 19676008 TI - [The relationship between emotional distress, cognitive performance and health - related quality of life in patients with hepatitis C prior to antiviral treatment]. AB - Recent epidemiologic studies suggest that more than 175 million individuals are infected with hepatitis C virus (HCV) worldwide. In the past few years, outcome studies in chronic HCV patients are no longer focusing solely on traditional end points such as mortality rates but on psychosocial well-being such as health related quality of life (HRQoL), emotional states, and neuropsychological functioning. The purpose of our exploratory study was to assess cross sectionally the frequency of depression, posttraumatic stress symptoms, cognitive deficits, and impairments in HRQoL in chronic HCV patients prior to antiviral treatment, and to investigate how cognitive impairments and emotional distress were related to quality of life. We recruited 34 chronic HCV patients who had presented for initial assessment of the need for antiviral therapy. Psychometric observer rating and self-rating scales were administered to evaluate posttraumatic stress symptoms (PTSS-10), depressive symptoms (BDI), HRQoL (SF-36 Health Status Questionnaire), and cognitive functioning (SKT). 32.4 % (n = 11) of the sample suffered from clinical depression, and 8.8 % (n = 3) had a posttraumatic stress syndrome. 8.8 % (n = 3) of the sample showed cognitive impairments. Significant impairments in HRQoL were found in the health-related domains vitality, role emotional, and role-physical. The severity of emotional distress as measured on the BDI and PTSS-10 was associated with decrements in HRQoL. However, lower cognitive function scores on the SKT were not associated with lower HRQoL SF-36 values. Chronic HCV patients seem to face a major risk of depression, posttraumatic stress symptoms, and cognitive dysfunction, and the presence of emotional distress is associated with impairments in quality of life. We therefore underscore the need for early and comprehensive bio-psycho-social diagnosis and therapy of chronic HCV patients in order to treat emotional distress and enhance patients; quality of life at an early stage before initiating antiviral therapy, as well as to expand the pool of patients eligible to receive antiviral therapy. PMID- 19676009 TI - [Medical product safety and patient safety--challenges at the "human-medical product" interface]. PMID- 19676010 TI - [BGH, Beschl. 6. May 2008, Az: VI ZR 250/07--from freestyle to responsibility]. PMID- 19676011 TI - [Guideline for the additional test positions according to the EPQC 4th Edition for Digital Mammography Systems]. AB - Within the physical-technical quality assurance of the German breast cancer screening program all digital mammography systems have to perform the contrast resolution test and the determination of the average glandular dose based on the European guidelines for quality assurance in breast cancer screening and diagnosis (4th Edition). Since 1.1.2009 this applies to digital systems outside the screening program too. To accomplish uniform measurements in all federal states of Germany, the physical board of the reference centers developed together a special guideline for these test position. This Guideline describes the determination of the average glandular dose for different types of mammography systems, the CDMAM image acquisition and the CDMAM image evaluation as well. This guideline was verified by the German task group "Rontgenverordnung". PMID- 19676012 TI - [Medical radiation exposure and justification at a large teaching hospital: comparison of radiation-related and disease-related risks]. AB - PURPOSE: The medical X-ray exposure was determined in a 2400-bed hospital. The radiation-related risk was compared with the severity of disease (ICD) to verify the justification for X-ray procedures. A model to estimate radiation and disease related "loss of lifetime" was applied. MATERIALS AND METHODS: X-ray exposure from radiography, fluoroscopy and CT was determined for diagnostic and interventional procedures during one hospital stay of 403 patients (0.5 % of all 80 000 patients/year). CTDI and DLP in CT, DAP in fluoroscopy or SED in radiography were used to calculate the effective dose (ED). The disease and radiation-related risk were compared with a simple "loss of lifetime" model. RESULTS: The mean age of all patients was 60. Only a subgroup of 170 patients (42 %) with a mean age of 67.6 had one or more X-ray procedures. The average ED of these exposed patients was 5.12 mSv. 14.4 % CT examinations had a dose contribution of 52.5 % followed by 5.3 % radiology and cardiology procedures at 37.2 %. 90 % of ED was applied to only 44 patients (11 %) and 50 % of ED was applied to only 10 patients (2.5 %) with a mean age of 71.2. 4 of these 10 patients had malignant tumors, the other 6 suffered from pulmonary embolism, pneumonia, stent-graft infection, CAD, Crohn's disease and severe hypertension. The ratio of disease vs. radiation-related risk was between 2.1 : 1 and 1800 : 1. CONCLUSION: In this study referral for justified X-ray procedures in a large hospital with high level medical care offers a significant patient benefit. PMID- 19676013 TI - [Results of an automatic evaluation of test images according to PAS 1054 and IEC 6220-1-2 on different types of digital mammographic units]. AB - AIM: To test whether image processing methods are suitable for detecting differences in the image quality of digital mammography units and whether the methods are more sensitive than visual test methods like the evaluation of CDMAM test images. METHODS: Test images according to PAS 1054 were acquired with CR and DR systems and analyzed automatically. Only parameters were used that are obligatory for calculating the detective quantum efficiency (DQE), the noise power spectrum (NPS), the spatial resolution (MTF, modulation transfer function), and the averaged glandular dose (AGD). The derived value, the NEQ index, is linked to the noise equivalent quanta (NEQ). Additionally CDMAM test images were acquired and evaluated at one CR and one DR system. RESULTS: The accuracy of the evaluated values is in the range below of 2 %. The accuracy requirements are fulfilled to establish values for detecting changes in image quality related to changes in dose no greater than one exposure value. If a visual evaluation of test images is used, a significant difference in image quality can only be detected when the dose is doubled. CONCLUSION: A method is proposed for establishing dose-correlated tolerances for values used in constancy and acceptance tests independently of the type of mammography unit (CR and DR systems). A method which abstains from visual tests for establishing the performance of digital mammography units could be used in acceptance tests. PMID- 19676014 TI - Delayed-enhancement magnetic resonance imaging in patients with clinically suspected stress cardiomyopathy (Tako-tsubo). AB - PURPOSE: To compare the ability of delayed-enhancement magnetic resonance imaging (DE-MRI) and other MRI and clinical parameters to identify diseases mimicking stress cardiomyopathy (SCM). MATERIALS AND METHODS: The study included 14 consecutive patients fulfilling the American Heart Association (AHA) criteria for SCM with acute left ventricular dysfunction in the absence of coronary artery disease, triggered by psychological stress. The MRI protocol consisted of cine, T 2-weighted, first-pass-perfusion (FPP) and DE-MRI. RESULTS: Six patients with DE were classified as mimicking SCM (non-SCM) and 8 patients without DE as SCM. FPP defects were found in 4 patients with non-SCM and in none with SCM (p < 0.05). Myocardial edema was found in 5 patients with non-SCM and in 2 patients with SCM (p = ns). No significant differences in clinical findings such as ECG, cardiac markers and echocardiographic recovery of left ventricular function were found between patients with non-SCM and SCM. CONCLUSION: Non-SCM defined by DE-MRI is a frequent finding in patients fulfilling the AHA criteria for SCM. Clinical findings seem to be of limited value to differentiate between non-SCM and SCM. PMID- 19676015 TI - [Image quality of CT angiography of coronary arteries depending on the degree of coronary calcification using a dual source CT scanner]. AB - PURPOSE: To evaluate the influence of severe calcification on image quality of CT angiography (CTA) of coronary arteries acquired with dual source CT. METHOD: 171 dual source CTAs in spiral mode were evaluated. Depending on the Agatston score (CS), patients where divided into four subgroups (I: CS 0 - 200, II: CS 201 - 600, III: CS 601 - 1000, IV: CS > 1000). Image quality was evaluated by two independent, blinded readers using a three-stage scale and the image quality of each subgroup was also shown with the help of an image quality index (BQI). In addition, a comparison with coronary catheter angiography was undertaken in a subgroup. RESULTS: Dual source CTA in spiral mode showed good image quality even in patients with Agatston scores of 601 - 1000 and > 1000. The four classes showed no significant difference in image quality. (BQI = 1,164 [I], BQI = 1,212 [II], BQI = 1,281 [III], BQI = 1,111 [IV]; p = 0,3145). The detailed analysis showed particularly good quality in patients with a heart rate < 75 bpm. The smaller subgroup of patients with CS > 200 and heart rates > 75 bpm showed a statistically significant reduction in image quality (BQI = 1,625; p = 0.0016). There were no gender differences in the image quality (p = 0.9881) although the calcium scores differed significantly in men (CS = 313 [+/- 449]) and women (CS = 145 [+/- 304]). The comparison with coronary catheter angiography showed a good correlation (92%). CONCLUSION: CTA of coronary arteries performed with dual source CT in spiral mode provides good image quality even in patients with higher calcification scores. The reduction of heart rates > 75 bpm improves the image quality significantly in patients with higher calcification scores. PMID- 19676016 TI - [Epulis--a rare congenital malformation]. PMID- 19676017 TI - [Empyema of the hip joint and psoas abscess: rare complications in inapparent sigmoid diverticulitis]. PMID- 19676018 TI - [MRI urography detection of an urinoma as a result of iatrogenic ureteral injury in roentgen contrast medium contraindications]. PMID- 19676019 TI - [Intestinal sarcoidosis--an unusual manifestation of a granulomatous systemic disease]. PMID- 19676020 TI - [Rosai-Dorfman disease with bilateral kidney involvement: morphologic image characteristics of a very rare organ manifestation]. PMID- 19676021 TI - [Image of an aorto-left ventricular tunnel in infectious endocarditis with dual source multidetector CT angiography]. PMID- 19676023 TI - The nasolabial fold as potential vascular receptor site: an anatomic study. AB - Free and pedicled flaps are frequently used in reconstruction of the lower two thirds of the face. For these reconstructions, the submandibular facial vessels are extensively used as a receptor site. In this anatomic study, we investigate if the facial vessels in the nasolabial fold can be used as a receptor site as well. In 13 human cadavers, the facial artery and vein were dissected in the nasolabial fold in the same way as would be done during surgery. The ease of dissection and length, diameter, and location of the vessels were analyzed. The average length of the dissected artery was 28 mm (+/-11 standard deviations [SD]) and of the dissected vein, 19 mm (+/-6 SD). The mean diameter of the artery was 1.5 mm (+/-0.4 SD) and 2.5 mm (+/-0.8 SD) for the vein. In 85% of the sides, both vessels were suitable to use as a microsurgical receptor site. The easy access and the measured diameter of the facial vessels in the nasolabial fold make it a potential site for microsurgical anastomosis. PMID- 19676024 TI - The influence of pedicle length on the viability of twisted perforator flaps in rats. AB - The pedicled perforator flap is now regarded as a valuable tool in the reconstructive armamentarium, but variable degrees of perforator twisting upon flap transposition are frequently encountered intraoperatively. The aim of this study was to investigate the effect of pedicle length on twisted perforator flap viability. Forty Sprague-Dawley rats were randomly divided into four groups of 10 animals each: group I (short pedicle, no rotation), group II (short pedicle, 720 degrees rotation), group III (long pedicle, no rotation), and group IV (long pedicle, 720 degrees rotation), and cranial epigastric artery perforator flaps were elevated. Mean group flap survival ratios were 88.47 +/- 13.87% (group I), 60.23 +/- 28.95% (group II), 87.06 +/- 13.85% (group III), and 84.28 +/- 31.76% (group IV). Viable flap ratios in group II were significantly lower than in groups I, III, and IV, respectively (p < 0.05). Our findings suggest that when perforator twisting is unavoidable, longer pedicles are significantly more likely to result in secure flap survival. PMID- 19676025 TI - [Workplace health promotion in network structures - the Erlangen Model of "enterprises in motion"]. AB - INTRODUCTION: Evidence-based health promotion and prevention are the only means to meet the future economic challenges in health care. Since preventive measures do not penetrate all strata of society alike, the workplace is a probable platform for health education and promotion. Against this background, the network of the 'Erlangen Model' attempts to include health promotion as an integral part of enterprise policy; the present paper evaluates preliminary results of this programme. METHOD: Questionnaires and interviews were conducted among employees of 6 companies and authorities forming the network "Agitating Enterprises". A total of 1,748 subjects were included and answered questions about their professional and health-related situation, physical activities, and expectations in connection with the programme. RESULTS: Almost half of the subjects (48%) had no intention to participate in one of the programme's courses. Most frequent mentioned reasons in favour of participation were the expectation of positive effects on general health (75%), well-being (78%), team work (32%) and enjoyment of sports (70%). Factor analysis extracted 5 dimensions of occupational burden out of over 50 items: "Co-operation with colleagues and superiors", "safety at work", "workflow organisation", "individual complaints" and "workplace design". Between participating companies the expression of these dimensions varied substantially; employees of the university hospital in general reported a higher than-average burden. In contrast, differences regarding the health status, satisfaction with employment conditions and individual activity scores were minor. CONCLUSION: Health promotion at the workplace is meaningful, especially for health-care employees. Differential analyses of reasons for non-participation may reveal starting points for an improvement of attendance in health-promotion programmes. PMID- 19676041 TI - Loss of STARD10 expression identifies a group of poor prognosis breast cancers independent of HER2/Neu and triple negative status. AB - The phospholipid transfer protein STARD10 cooperates with c-erbB signaling and is overexpressed in Neu/ErbB2 breast cancers. We investigated if STARD10 expression provides additional prognostic information to HER2/neu status in primary breast cancer. A published gene expression dataset was used to determine relationships between STARD10 and HER2 mRNA levels and patient outcome. The central findings were independently validated by immunohistochemistry in a retrospective cohort of 222 patients with breast cancer with a median follow-up of 64 months. Kaplan Meier and Cox proportional hazards analyses were used for univariate and multivariate analyses. Patients with low STARD10 or high HER2 tumor mRNA levels formed discrete groups each associated with a poor disease-specific survival (p = 0.0001 and p = 0.0058, respectively). In the immunohistochemical study low/absent STARD10 expression i.e. < or = 10% positive cells was observed in 24 of 222 (11%) tumors. In a univariate model, low/absent STARD10 expression was significantly associated with decreased patient survival (p = 0.0008). In multivariate analyses incorporating tumor size, tumor grade, lymph node status, ER, PR and HER2 status, low STARD10 expression was an independent predictor of death from breast cancer (HR: 2.56 (95% CI: 1.27-5.18), p = 0.0086). Furthermore, low/absent STARD10 expression, HER2 amplification and triple negative status were independent prognostic variables. Loss of STARD10 expression may provide an additional marker of poor outcome in breast cancer identifying a subgroup of patients with a particularly adverse prognosis, which is independent of HER2 amplification and the triple negative phenotype. PMID- 19676042 TI - BIIB021, a synthetic Hsp90 inhibitor, has broad application against tumors with acquired multidrug resistance. AB - 17-AAG, the first-generation clinical Hsp90 inhibitor, exhibits promising antitumor activity in clinical studies, but is limited by poor solubility and hepatotoxicity. To pursue compounds with better biopharmaceutical properties, we have developed a series of fully synthetic orally bioavailable inhibitors of Hsp90. Here, we report that 17-AAG and other ansamycin derivatives are inactive in P-gp and/or MRP-1 expressing cell lines and sensitivity could be restored by coadministration of P-gp or MRP inhibitors. In contrast, the synthetic Hsp90 inhibitor, BIIB021 was active in these models. Accordingly, BIIB021 was considerably more active than 17-AAG against adrenocortical carcinoma, a tumor that naturally expresses P-gp, both in vitro and in vivo. This efflux pump mediated resistance is manifested in both cytotoxicity assays and measurements of target inhibition, such as client protein degradation. Other than this, the cytotoxic activity of BIIB021 was also not influenced by loss of NQO1 or Bcl-2 overexpression, molecular lesions that do not prevent client loss but are nonetheless associated with reduced cell killing by 17-AAG. Our results indicate that the activity of 17-AAG and other ansamycins may be curtailed in tumors that have upregulated efflux pumps or antiapoptotic proteins or other genetic alterations. These data indicate that the new generation of synthetic anti-Hsp90 drugs, exemplified by BIIB021 that is currently undergoing Phase II testing, may have broader application against tumors with acquired multidrug resistance or tumors located in organs protected by MDR proteins, such as the adrenal glands, brain and testis. PMID- 19676043 TI - Downregulations of B-cell lymphoma 2 and myeloid cell leukemia sequence 1 by microRNA 153 induce apoptosis in a glioblastoma cell line DBTRG-05MG. AB - MicroRNA-153 (miR-153) is a brain-specific miRNA that is expressed at a significantly lower level in glioblastoma (GBM) relative to non-neoplastic brain tissue. Although the expression pattern of miR-153 has been extensively established, its target genes and cellular mechanism remain undefined. To investigate into the potential function of miR-153 in glioblastmas, we transfected a GBM cell line DBTRG-05MG with synthetic miR-153 oligos and observed decreased cell proliferation and increased apoptosis. Bioinformatics analysis revealed that anti-apoptosis family member B-cell lymphoma 2 (Bcl-2) and myeloid cell leukemia sequence 1 (Mcl-1) are potential targets of miR-153. Indeed, Western blot analysis indicated that miR-153 downregulated both Bcl-2 and Mcl-1 at the protein levels. Single strand miR-153 inhibitor, which forms complementary base-pair with endogenous miR-153, efficiently blocked the apoptosis and target protein degradation induced by overexpression of miR-153. By luciferase reporter assays, we further showed that miR-153 inhibited Bcl-2 and Mcl-1 expressions by directly targeting the 3'UTR regions of their respective mRNAs. PMID- 19676044 TI - CD133 is indicative for a resistance phenotype but does not represent a prognostic marker for survival of non-small cell lung cancer patients. AB - Despite advances in anticancer treatment, lung cancer still has poor prognosis. Recently, a cancer stem cell (CSC) hypothesis has emerged describing a small subset of tumor cells with stem cell properties. CSCs found in many solid tumors express CD133 antigen on the cell surface. The presence of CSC is correlated with poor survival of patients with glioblastomas, colon or prostate cancers. In this study, we evaluated whether CD133 expression in non-small cell lung cancer (NSCLC) has a prognostic value in patients' survival. We also analyzed whether CD133 positivity of NSCLC correlates with the expression of resistance-related proteins, angiogenic factors, oncogenes, proliferative activity or apoptosis. CD133 expression was retrospectively examined in a total of 88 cases of previously untreated NSCLC by immunohistochemistry. We found no correlation between CD133 positivity or the amount of CD133(+) cells with NSCLC patients' survival, expression of oncogenes c-myc, c-N-ras, c-jun, c-fos, c-erbB1, c-erbB2 or p53, angiogenic factors VEGF, VEGFR-1, FGF, FGFR-1, tissue factor and with proliferative activity or apoptosis in NSCLC tissues. However, there was a significant association between the expression of resistance-related proteins glutathione S-transferase, thymidylate synthase, catalase, O(6)-methylguanine-DNA methyltransferase and p170 and CD133. Because CD133 expression is linked to a resistant phenotype, detection of CD133(+) cells may be useful to predict efficacy of cytotoxic therapy but CD133 is not a strong prognostic parameter for survival of patients with NSCLC. PMID- 19676045 TI - Diagnostic and prognostic implications of microRNA profiling in prostate carcinoma. AB - This study aimed to investigate the microRNA (miRNA) profile in prostate carcinoma tissue by microarray analysis and RT-qPCR, to clarify associations of miRNA expression with clinicopathologic data and to evaluate the potential of miRNAs as diagnostic and prognostic markers. Matched tumor and adjacent normal tissues were obtained from 76 radical prostatectomy specimens. Twenty-four tissue pairs were analyzed using human miRNA microarrays for 470 human miRNAs. Differentially expressed miRNAs were validated by TaqMan RT-qPCR using all 76 tissue pairs. The diagnostic potential of miRNAs was calculated by receiver operating characteristics analyses. The prognostic value was assessed in terms of biochemical recurrence using Kaplan-Meier and Cox regression analyses. Fifteen differentially expressed miRNAs were identified with concordant fold-changes by microarray and RT-qPCR analyses. Ten microRNAs (hsa-miR-16, hsa-miR-31, hsa-miR 125b, hsa-miR-145, hsa-miR-149, hsa-miR-181b, hsa-miR-184, hsa-miR-205, hsa-miR 221, hsa-miR-222) were downregulated and 5 miRNAs (hsa-miR-96, hsa-miR-182, hsa miR-182, hsa-miR-183, hsa-375) were upregulated. Expression of 5 miRNAs correlated with Gleason score or pathological tumor stage. Already 2 microRNAs classified up to 84% of malignant and nonmalignant samples correctly. Expression of hsa-miR-96 was associated with cancer recurrence after radical prostatectomy and that prognostic information was confirmed by an independent tumor sample set from 79 patients. That was shown with hsa-miR-96 and the Gleason score as final variables in the Cox models build in the 2 patient sets investigated. Thus, differential miRNAs in prostate cancer are useful diagnostic and prognostic indicators. This study provides a solid basis for further functional analyses of miRNAs in prostate cancer. PMID- 19676046 TI - Prognostic significance of integrin-linked kinase1 overexpression in astrocytoma. AB - Integrin-linked kinase 1 (ILK1), a member of the serine/threonine kinases, has been demonstrated to be associated with numerous biological and pathological processes. However, the clinical and functional significance of ILK1 expression has not been characterized previously in human astrocytoma. In this study, we found that ILK1 was overexpressed, at both mRNA and protein levels, in astrocytoma cell lines as compared with normal human astrocytes. The ILK1 mRNA and protein were significantly increased up to 5.6-fold and 10.1-fold, respectively, in primary astrocytoma in comparison with the paired adjacent noncancerous brain tissues obtained from the same patient. Furthermore, immunohistochemical analysis revealed that ILK1 protein was positive in 208 of 228 (91.2%) paraffin-embedded archival astrocytoma specimens. Statistical analysis suggested that the upregulation of ILK1 was significantly correlated with the histological grading of astrocytoma (p = 0.000), and that patients with high ILK1 level exhibited shorter survival time (p < 0.001). Multivariate analysis revealed that ILK1 upregulation might be an independent prognostic indicator for the survival of patients with astrocytoma. Taken together, our results suggest that ILK1 might represent a novel and useful prognostic marker for astrocytoma and play a role during the development and progression of the disease. PMID- 19676047 TI - Optimisation of enzyme assisted extraction of silybin from the seeds of Silybum marianum by Box-Behnken experimental design. AB - INTRODUCTION: Silybin, a standardised extract of flavanolignans from the seeds of Silybum marianum, has been used for centuries as a natural remedy in the treatment of hepatitis and cirrhosis. The higher yield of silybin by using more efficient extraction technique is of particular interest in the herbal products manufacture. OBJECTIVE: To systematically investigate the important factors of enzyme-assisted extraction of flavanolignans from the seeds of Silybum marianum to enhance the extraction yield of silybin. METHODOLOGY: The important factors of enzyme-assisted extraction were optimised by employing Box-Behnken design with the aid of the orthogonal array design (OAD) OA(8) (2(7)). The effects of enzyme incubation temperature (EIT), the pH of enzyme solution (PES) and the size of seeds (SS) on the yield of silybin were visualised as three-dimensional response surface and contour plots. RESULTS: The predictive yield was 24.6 mg/g defatted seeds under the optimum enzymolysis conditions (EIT 40 degrees C, PES 4.5 and SS 7003 microm). The coefficient of the model was r(2) > 0.97 (n = 15). The actual yield of silybin was 24.81 + or - 1.93 mg/g defatted seeds, higher by 138 and 123.6% than that from ethanol extraction in this study and in the previous literature, respectively. IR spectra and HPLC of the extracts by EAE were in agreement with those from ethanol extraction. SEM and TEM pictures of defatted seeds by variant extractions demonstrate that the extraction of silybin depends on the destruction of cell walls. CONCLUSION: The results suggest that EAE is a promising alternative for the extraction of silybin by the use of traditional ethanol extraction. PMID- 19676048 TI - Assessing trade in health services in countries of the Eastern Mediterranean from a public health perspective. AB - SUMMARY: Assessing trade in health services (TiHS) in developing countries is challenging since the sources of information are diverse, information is not accessible and professionals lack grasp of issues. A multi-country study was conducted in the Eastern Mediterranean Region (EMR)--Egypt, Jordan, Lebanon, Morocco, Oman, Pakistan, Sudan, Syrian Arab Republic, Tunisia, and Yemen. The objective was to estimate the direction, volume, and value of TiHS; analyze country commitments; and assess the challenges and opportunities for health services.Trade liberalization favored an open trade regime and encouraged foreign direct investment. Consumption abroad and movement of natural persons were the two prevalent modes. Yemen and Sudan are net importers, while Jordan promotes health tourism. In 2002, Yemenis spent US$ 80 million out of pocket for treatment abroad, while Jordan generated US$ 620 million. Egypt, Pakistan, Sudan and Tunisia export health workers, while Oman relies on import and 40% of its workforce is non-Omani. There is a general lack of coherence between Ministries of Trade and Health in formulating policies on TiHS.This is the first organized attempt to look at TiHS in the EMR. The systematic approach has helped create greater awareness, and a move towards better policy coherence in the area of trade in health services. PMID- 19676049 TI - Re: coffee drinking and risk of endometrial cancer--a population-based cohort study. PMID- 19676050 TI - Detailed examination of lymph nodes improves prognostication in colorectal cancer. AB - Up to 30% of stage II patients with curatively resected colorectal cancer (CRC) will develop disease recurrence. We evaluated whether examination of lymph nodes by multilevel sectioning and immunohistochemical staining can improve prognostication. Lymph nodes (n = 780) from 36 CRC patients who had developed disease recurrence (cases) and 72 patients who showed no recurrence of disease for at least 5 years (controls) were analyzed. Sections of 4 levels at 200-microm interval were immunohistochemically stained for cytokeratin expression. The first level was analyzed by conventional and automated microscopy, and the 3 following levels were analyzed by automated microscopy for the presence of tumor cells. Overall, cases showed more micrometastases (3 patients) than controls (1 patient). Analysis of a second level led to the additional detection of 1 patient with micrometastases (case) and 1 patient with macrometastasis (case). Examining more levels only led to additional isolated tumor cells, which were equally divided between cases and controls. Likewise, automated microscopy resulted only in detection of additional isolated tumor cells when compared with conventional microscopy. In multivariate analysis, micrometastases [odds ratio (OR) 26.3, 95% confidence interval (CI) 1.9-364.8, p = 0.015], T4 stage (OR 4.8, 95% CI 1.4 16.7, p = 0.013) and number of lymph nodes (OR 0.9, 95% CI 0.8-1.0, p = 0.028) were independent predictors for disease recurrence. Lymph node analysis of 2 levels and immunohistochemical staining add to the detection of macrometastases and micrometastases in CRC. Micrometastases were found to be an independent predictor of disease recurrence. Isolated tumor cells were of no prognostic significance. PMID- 19676051 TI - Sensitization of ovarian carcinoma cells to the atypical retinoid ST1926 by the histone deacetylase inhibitor, RC307: enhanced DNA damage response. AB - The synthetic atypical retinoids containing an adamantyl group exhibit antiproliferative or proapoptotic activities. Apoptosis induction is a dose dependent effect independent of retinoid receptors. We have reported that induction of apoptosis by the atypical retinoid, ST1926, is associated with early manifestations of genotoxic stress. Indeed, in this study performed in ovarian carcinoma cells, we show that exposure to ST1926 resulted in an increase of early markers of DNA damage, including ATM and H2AX phosphorylation. In addition, we found that a novel histone deacetylase (HDAC) inhibitor (RC307) was able to enhance sensitivity of ovarian carcinoma cells to ST1926. Under conditions where single-agent treatment caused only antiproliferative effects, the combination of the atypical retinoid and HDAC inhibitor resulted in marked apoptotic cell death with a more rapid onset in wild-type p53 ovarian carcinoma cells. The sensitization to ST1926-induced apoptosis was associated with an enhanced DNA damage response, because a prolonged expression of DNA damage markers (e.g., H2AX, p53 and RPA-2 phosphorylation) and a marked activation of DNA damage checkpoint kinases (in particular, phosphorylation of Chk1) were observed indicating an accumulation of DNA damage by the ST1926/HDAC inhibitor combination. The study provides additional support to the role of DNA damage as a primary event leading to the activation of apoptosis in ovarian carcinoma cells by adamantyl retinoids and documents the potential therapeutic efficacy of the combination of ST1926 and HDAC inhibitors of the novel series. PMID- 19676052 TI - Small molecule XIAP inhibitors sensitize childhood acute leukemia cells for CD95 induced apoptosis. AB - Escape of apoptosis may contribute to treatment failure in childhood acute lymphoblastic leukemia (ALL) calling for new approaches to overcome apoptosis resistance. Here, we provide for the first time evidence that small molecule inhibitors that target the anti-apoptotic protein X-linked inhibitor of apoptosis (XIAP) sensitize ALL cells for CD95-induced apoptosis. XIAP inhibitors at subtoxic concentrations, but not a structurally related control compound, act synergistically with agonistic anti-CD95 antibodies or MegaFasL, a hexameric form of CD95 ligand, to induce apoptosis in ALL cells. Further, XIAP inhibitors co operate with MegaFasL to reduce clonogenic survival of ALL cells demonstrating their effect also on long-term survival. In contrast, XIAP inhibitors show little effect on MegaFasL-mediated apoptosis in normal peripheral blood lymphocytes (PBLs), pointing to some tumor selectivity. Molecular studies reveal that XIAP inhibitors enhance CD95-induced activation of caspases, loss of mitochondrial membrane potential and cytochrome c release in a caspase-dependent manner. Importantly, XIAP inhibitors sensitize primary leukemic blasts from children with ALL for MegaFasL-induced apoptosis. Thus, small molecule XIAP inhibitors present a promising novel approach to enhance CD95-induced apoptosis in childhood acute leukemia. PMID- 19676053 TI - Targeted knockdown of Bcl2 in tumor cells using a synthetic TRAIL 3'-UTR microRNA. AB - Targeting tumor-related overexpression of anti-apoptotic Bcl2 protein by RNAi has been suggested as a potential treatment for cancer. However, the stability of RNAi and its delivery are still major obstacles to the clinical testing of Bcl2 RNAi. Here, we explore a novel strategy of expressing a synthetic Bcl2 microRNA (smRNA) in the 3' untranslated region (UTR) of the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), an apoptosis-inducing protein without apparent toxic effects in normal cells. TRAIL was specifically expressed from the human telomerase reverse transcriptase promoter (pTRT) that is active in many human tumors. Using this approach, we demonstrated that pTRT drove the tumor-specific expression of Bcl2 smRNA, which was processed by the host RNAi machinery and silenced endogenous Bcl2 expression in tumor cells. Bcl2 smRNA induced tumor cell apoptosis by activating caspase-3 and led to significant sensitization of tumor cells to TRAIL-induced apoptosis, while normal cells were spared. We also showed that the combined therapy of TRAIL-induced apoptosis and Bcl2 downregulation was superior to the mono-therapy of TRAIL or Bcl2 smRNA alone. This study proves a general paradigm for cancer therapy by using 3' UTR microRNA technology. PMID- 19676054 TI - Udp-glucose dehydrogenase as a novel field-specific candidate biomarker of prostate cancer. AB - Uridine diphosphate (UDP)-glucose dehydrogenase (UGDH) catalyzes the oxidation of UDP-glucose to yield UDP-glucuronic acid, a precursor for synthesis of glycosaminoglycans and proteoglycans that promote aggressive prostate cancer (PC) progression. The purpose of our study was to determine if the UGDH expression in normal appearing acini (NAA) from cancerous glands is a candidate biomarker for PC field disease/effect assayed by quantitative fluorescence imaging analysis (QFIA). A polyclonal antibody to UGDH was titrated to saturation binding and fluorescent microscopic images acquired from fixed, paraffin-embedded tissue slices were quantitatively analyzed. Specificity of the assay was confirmed by Western blot analysis and competitive inhibition of tissue labeling with the recombinant UGDH. Reproducibility of the UGDH measurements was high within and across analytical runs. Quantification of UGDH by QFIA and Reverse-Phase Protein Array analysis were strongly correlated (r = 0.97), validating the QFIA measurements. Analysis of cancerous acini (CA) and NAA from PC patients vs. normal acini (NA) from noncancerous controls (32 matched pairs) revealed significant (p < 0.01) differences, with CA (increased) vs. NA, NAA (decreased) vs. NA and CA (increased) vs. NAA. Areas under the Receiver Operating Characteristic curves were 0.68 (95% CI: 0.59-0.83) for NAA and 0.71 (95% CI: 0.59-0.83) for CA (both vs. NA). These results support the UGDH content in prostatic acini as a novel candidate biomarker that may complement the development of a multi-biomarker panel for detecting PC within the tumor adjacent field on a histologically normal biopsy specimen. PMID- 19676055 TI - Detection of parent-of-origin effects using general pedigree data. AB - Genomic imprinting is an important epigenetic factor in complex traits study, which has generally been examined by testing for parent-of-origin effects of alleles. For a diallelic marker locus, the parental-asymmetry test (PAT) based on case-parents trios and its extensions to incomplete nuclear families (1-PAT and C PAT) are simple and powerful for detecting parent-of-origin effects. However, these methods are suitable only for nuclear families and thus are not amenable to general pedigree data. Use of data from extended pedigrees, if available, may lead to more powerful methods than randomly selecting one two-generation nuclear family from each pedigree. In this study, we extend PAT to accommodate general pedigree data by proposing the pedigree PAT (PPAT) statistic, which uses all informative family trios from pedigrees. To fully utilize pedigrees with some missing genotypes, we further develop the Monte Carlo (MC) PPAT (MCPPAT) statistic based on MC sampling and estimation. Extensive simulations were carried out to evaluate the performance of the proposed methods. Under the assumption that the pedigrees and their associated affection patterns are randomly drawn from a population of pedigrees with at least one affected offspring, we demonstrated that MCPPAT is a valid test for parent-of-origin effects in the presence of association. Further, MCPPAT is much more powerful compared to PAT for trios or even PPAT for all informative family trios from the same pedigrees if there is missing data. Application of the proposed methods to a rheumatoid arthritis dataset further demonstrates the advantage of MCPPAT. PMID- 19676056 TI - Microdeletion syndrome 16p11.2-p12.2: clinical and molecular characterization. AB - The pericentromeric region on 16p appears to be susceptible to chromosomal rearrangements and several patients with rearrangements in this region have been described. We report on a further patient with a microdeletion 16p11.2-p12.2 in the context of described patients with a deletion in the pericentromeric region of 16p. Minor facial anomalies, feeding difficulties, significant delay in speech development, and recurrent ear infections are common symptoms of the microdeletion syndrome 16p11.2-p12.2. All reported patients so far share a common distal breakpoint at 16p12.2 but vary in the proximal breakpoint at 16p11.2. The microdeletion 16p11.2-p12.2 should be distinguished from the approximately 500 kb microdeletion in 16p11.2 which seems to be associated with autism but not with facial manifestations, feeding difficulties, or developmental delay. PMID- 19676057 TI - Clinical lessons learned from the International Research Symposium on Ankyloblepharon-Ectodermal Defects-Cleft Lip/Palate (AEC) syndrome. AB - The International Research Symposium on Ankyloblepharon-Ectodermal Defects-Cleft Lip/Palate ((AEC) Syndrome, that was supported by the National Foundation for Ectodermal Dysplasias (NFED) through a grant from the National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS) and the National Institutes of Health Office of Rare Diseases (NIH-ORD), brought together physicians, scientists, and 23 individuals affected by AEC syndrome from 13 families. Eighteen of the AEC-affected individuals were enrolled in an IRB approved protocol through Baylor College of Medicine. Enrolled participants had clinical evaluations by multiple subspecialists, and additionally submitted blood for mutational analysis and skin specimens for pathologic evaluation. One of the goals of the conference was to define clinical and pathologic findings for improved diagnostic criteria, with the hope of determining genotype-phenotype correlations that might aid in predicting prognosis or directing therapeutics. What we found was wide interfamilial and intrafamilial variability in the manifestations of the syndrome. We were unable to identify any specific genotype phenotype correlations. This may relate to our small sample size or other unknown epigenetic factors that are also at play in the expression and manifestation of the syndrome in specific individuals. PMID- 19676058 TI - Growth, nutritional, and gastrointestinal aspects of ankyloblepharon-ectodermal defect-cleft lip and/or palate (AEC) syndrome. AB - Ankyloblepharon-ectodermal defect-cleft lip and/or palate (AEC) is a rare genetic disorder due to mutations in the TP63 gene. In the present study, we characterized the pattern of growth and body composition and the nutritional and gastrointestinal aspects of children and adults (n = 18) affected with this disorder using clinical anthropometry and a survey questionnaire. The mean birth weight and height-for-age z-scores of the AEC patients were significantly lower than those of the reference population. The weight-for-age z-score of the AEC cohort increased significantly with advancing age because of increasing body fat. Cleft lip and palate were present in 47% and 94%, respectively, of the AEC cohort; 28% had dentures. One-fourth or more of the AEC cohort reported having nutritional and/or gastrointestinal problems including the need for supplemental formula feedings, gastrostomy placement, gastroesophageal reflux, and constipation. Our observations provide novel clinical information about growth, body composition, and nutritional and gastrointestinal aspects of children and adults with AEC. PMID- 19676059 TI - Craniofacial and anthropometric phenotype in ankyloblepharon-ectodermal defects cleft lip/palate syndrome (Hay-Wells syndrome) in a cohort of 17 patients. AB - Ankyloblepharon-ectodermal dysplasia-cleft lip/palate (AEC) syndrome and Rapp Hodgkin syndrome are well-characterized clinical entities caused by mutations in the TP63 gene. While AEC and Rapp-Hodgkin had been thought to be clinically distinct entities, the elucidation of their molecular etiology confirmed that they are a clinical continuum as opposed to distinct disorders. We have evaluated 17 patients with AEC syndrome using a systematic clinical approach. In our study, we have identified new features and others that were thought to occur only rarely. These include short stature and poor weight gain with preservation of head circumference in nearly all subjects, trismus in 35% and hypospadias in 78% of males. In addition, we describe the frequency of phenotypic features and demonstrate the extreme clinical variability in the largest cohort of AEC individuals reported in the literature thus far. PMID- 19676060 TI - Spectrum of p63 mutations in a selected patient cohort affected with ankyloblepharon-ectodermal defects-cleft lip/palate syndrome (AEC). AB - Heterozygous mutations in the p63 gene underlie a group of at least seven allelic syndromes, including ankyloblepharon-ectodermal defects-cleft lip/palate syndrome (AEC) and Rapp Hodgkin syndrome (RHS), which involves varying degrees of ectodermal dysplasia, orofacial clefting and limb malformations. Mutations in the AEC and Rapp Hodgkin syndromes cluster in the 3' end of the p63 gene. Previously reported mutations are mainly missense and frameshift mutations in exons 13 and 14, affecting the p63alpha-specific SAM (sterile alpha motif) and TI (transactivation inhibitory) domains. A patient cohort affected by AEC syndrome was evaluated during International Research Symposium supported by the National Foundation for Ectodermal Dysplasias. Nineteen patients underwent full clinical evaluations and 18 had findings consistent with a diagnosis of AEC syndrome. These 19 patients, along with 5 additional relatives had genomic DNA analysis. Twenty-one of the 24 participants from 12 families were found to have mutations in the p63 gene. Eleven different mutations were identified; 10 were novel mutations. Eight were missense mutations within the coding region of the SAM domain. Three other mutations were located in exon 14 sequences, which encode the TI domain. The effects of the mutations in the SAM and TI domains are poorly understood and functional studies are required to understand the pathological mechanisms. However, AEC and RHS mutations in the 5' and 3' ends of the p63 gene point towards a critical role of the DeltaNp63alpha isoform for the AEC/RHS phenotype. PMID- 19676066 TI - Endometrial blood flow is impaired in women with polycystic ovarian syndrome who are clinically hyperandrogenic. AB - OBJECTIVES: To quantify endometrial and subendometrial blood flow in Caucasian women with polycystic ovarian syndrome (PCOS) and to determine whether these values differ according to the phenotypic expression of PCOS. METHODS: Transvaginal pelvic ultrasound was performed on the 3(rd)-5(th) day of the menstrual cycle in 36 women with PCOS and 36 controls to examine the endometrial and subendometrial vascularity. The subendometrial and endometrial blood flow indices (vascularizaton index (VI), flow index (FI) and vascularization flow index (VFI)) were measured using three-dimensional power Doppler angiography. Uterine artery blood flow was assessed through analysis of two-dimensional (2D) pulsed-wave Doppler waveforms. Analysis was performed to compare PCOS with non PCOS women, and subgroup analysis was performed of the PCOS women categorized according to their phenotypic manifestation. RESULTS: There were no significant differences in endometrial volume, subendometrial vascularity and uterine artery blood flow between women with PCOS and controls after controlling for body mass index (BMI). On subgroup analysis, compared with anovulatory but clinically normoandrogenic women with polycystic ovaries (PCO) and with controls, women with PCO who were both clinically hyperandrogenic and anovulatory had significantly lower endometrial (VI: 0.57% vs. 1.11% and 0.86%, respectively, both P = 0.01; VFI: 0.14 vs. 0.42 and 0.28, respectively, both P = 0.02) and subendometrial (VI: 1.59% vs. 3.17% and 2.47%, P = 0.01 and 0.02, respectively; VFI: 0.50 vs. 1.67 and 0.96, P = 0.01 and 0.02, respectively) blood flow. Moreover, clinically hyperandrogenic but ovulatory women with PCO also had significantly lower endometrial blood flow (VI: 0.52% vs. 1.11%, P = 0.04) than did anovulatory but clinically normoandrogenic women with PCO. There were no differences in any of the 2D pulsed-wave Doppler measures of blood flow between the subgroups. CONCLUSIONS: Subendometrial and endometrial blood flow is significantly impaired in women with PCOS who have clinical signs of hyperandrogenism. PMID- 19676072 TI - TIM-3 is expressed on activated human CD4+ T cells and regulates Th1 and Th17 cytokines. AB - TIM-3 is a molecule selectively expressed on a subset of murine IFN-gamma secreting T helper 1 (Th1) cells but not Th2 cells, and regulates Th1 immunity and tolerance in vivo. At this time little is known about the role of TIM-3 on human T cells. To determine if TIM-3 similarly identifies and regulates Th1 cells in humans, we generated a panel of mAb specific for human TIM-3. We report that TIM-3 is expressed by a subset of activated CD4(+) cells, and that anti-CD3/anti CD28 stimulation increases both the level of expression as well as the number of TIM-3(+) T cells. We also find that TIM-3 is expressed at high levels on in vitro polarized Th1 cells, and is expressed at lower levels on Th17 cells. In addition, human CD4(+) T cells secreted elevated levels of IFN-gamma, IL-17, IL-2, and IL 6, but not IL-10, IL-4, or TNF-alpha, when stimulated with anti-CD3/anti-CD28 in the presence of TIM-3-specific, putative antagonistic antibodies. This was not mediated by differences in proliferation or cell death, but rather by induction of cytokines at the transcriptional level. These results suggest that TIM-3 is a negative regulator of human T cells and regulates Th1 and Th17 cytokine secretion. PMID- 19676073 TI - CD137 ligand-mediated reverse signals increase cell viability and cytokine expression in murine myeloid cells: involvement of mTOR/p70S6 kinase and Akt. AB - Cross-linking of CD137 ligand (CD137L), a member of the TNF family, with recombinant CD137-Fc (rCD137-Fc) protein enhanced adherence of bone marrow derived macrophages, and increased the expression of ICAM-1, IL-1beta, IL-6, M CSF and phosphotyrosine proteins. In RAW264.7 cells, a murine myeloid cell line, rCD137-Fc not only increased adherence but also cell multiplication, in a manner comparable to LPS or M-CSF. In addition, it up-regulated expression of IL-1beta, IL-1 receptor antagonist, IL-6, COX2, tenascin C, neuropeptide Y and M-CSF mRNA. Neutralization of M-CSF by incubating the RAW264.7 cells with anti-M-CSF mAb did not prevent the CD137L signal-induced viability. Viability was blocked by PP2, an Src tyrosine kinase inhibitor, rapamycin, an mTOR inhibitor and LY294002, a PI3K inhibitor, but not by Wortmannin, another PI3K inhibitor. Cross-linking of CD137L increased phosphorylation of Akt and p70S6 kinase. The latter was blocked by PP2, rapamycin or LY294002, but not by Wortmannin, whereas phosphorylation of Akt was blocked by LY294002 or Wortmannin. These findings demonstrate that reverse signals evoked by CD137L regulate immune functions in macrophages. PMID- 19676074 TI - Tissue transglutaminase enhances collagen type II-induced arthritis and modifies the immunodominant T-cell epitope CII260-270. AB - The calcium-dependent enzyme tissue transglutaminase (tTG) is associated with diverse biological functions, such as induction of apoptosis, modeling of the extracellular matrix, receptor-mediated endocytosis, cell growth and differentiation, cell adhesion and signal transduction. Also, it may deamidate glutamine residues to glutamic acid and catalyze cross-linking of proteins. In this study, we have investigated the impact of tTG for posttranslational modifications and cross-linking of the immunodominant T-cell epitope CII260-270 and their effects on the collagen-induced arthritis, an animal model for rheumatoid arthritis. By using mass spectrometry analysis and hybridoma assays, we have demonstrated that tTG could perform both types of modifications (deamidation and cross-link formation) on the immunodominant T-cell epitope CII259-273. Replacement of the glutamine at position 267 with glutamic acid leads to a decreased binding affinity to MHC II. T cells recognized both non-modfied (Q(267)) and modified (E(267)) CII259-273-peptides. We also show that administration of tTG leads to increased incidence, severity and histopathological manifestations of collagen-induced arthritis in mice. Moreover, we conclude that both processes, deamidation and cross-linking, are involved in the tTG-catalyzed reactions, and in vivo administration of tTG enhances arthritis severity and joint destruction in mice. PMID- 19676075 TI - Stable silencing of the glucocorticoid receptor in myelin-specific T effector cells by retroviral delivery of shRNA: insight into neuroinflammatory disease. AB - Autoimmune responses in the CNS can be induced by adoptive transfer of CD4(+) T effector cells after antigen-restimulation and expansion of clonal cell lines in vitro. However, pathogenic factors remain partially elusive due to the lack of appropriate methods to achieve gene inactivation. Here we describe a protocol for stable gene silencing in differentiated rat T cells by retroviral transfer of small hairpin RNAs. Through the combination of an expression cassette containing the green fluorescent protein with a puromycin selection cassette this allows for the generation of pure knockdown cell lines suitable for tracking in animals. Exemplified for the glucocorticoid receptor, we demonstrate that gene silencing renders T effector cells unresponsive to ligand-induced apoptosis and gene regulation without affecting their ability to induce EAE in rats. Interestingly, glucocorticoid administration remains effective in the treatment of EAE despite strongly diminished glucocorticoid receptor expression in antigen-specific T cells. This highlights an important role of other cell types and bystander T cells as targets of glucocorticoid therapy. Collectively, our approach provides a simple tool for stable and efficient gene silencing in T effector cells, which should help to better understand brain autoimmune pathophysiology. PMID- 19676076 TI - Electrostatic co-assembly of magnetic nanoparticles and fluorescent nanospheres: a versatile approach towards bimodal nanorods. PMID- 19676077 TI - Comment on "The emerging roles of cystatins in Alzheimer's disease". DOI 10.1002/bies.200900012. PMID- 19676080 TI - False impressions, intuition and the nature of methods: how insufficient study of methods confounds the study of biology itself. PMID- 19676081 TI - Effects of the 5 alpha-reductase inhibitor dutasteride on gene expression in prostate cancer xenografts. AB - BACKGROUND: In the prostate, androgens play a crucial role in normal and cancerous growth; hence the androgenic pathway has become a target of therapeutic intervention. Dutasteride is a 5 alpha-reductase (5AR) inhibitor currently being evaluated both for chemoprevention and treatment of prostate cancer. Dutasteride inhibits both 5AR I and II enzymes, effectively blocking conversion of testosterone to dihydrotestosterone (DHT) in the prostate. This greatly reduces the amount of the active ligand DHT available for binding to the androgen receptor (AR) and stimulating proliferation, making this a good candidate for chemoprevention of prostate cancer. In this study, we sought to determine how dutasteride is functioning at the molecular level, using a prostate cancer xenograft model. METHODS: Androgen-responsive LuCaP 35 xenograft tumors were grown in Balb/c mice. Subcutaneously implanted time-release pellets were used for drug delivery. Microarray analysis was performed using the Affymetrix HG-U133Av2 platform to examine changes in gene expression in tumors following dutasteride treatment. RESULTS: Dutasteride significantly reduced tumor growth in LuCaP 35 xenografts by affecting genes involved in apoptotic, cytoskeletal remodeling, and cell cycle pathways among others. Notably, genes in the Rho GTPase signaling pathway, shown to be important in androgen-deprivation conditions, were significantly up-regulated. CONCLUSION: We have identified multiple pathways outside of the androgenic pathway in prostate cancer xenografts affected by treatment with dutasteride. These findings provide insights into the function of dutasteride within the tumor microenvironment, potentially allowing for development of agents that can be used in combination with this drug to further enhance its effectiveness. PMID- 19676082 TI - The protective effect of diabetes mellitus against prostate cancer: role of sex hormones. AB - BACKGROUND: Diabetes mellitus (DM) has been associated with decreased risk of prostate cancer (PC) in several reports. Hormonal environment of diabetic patients is believed to be an important contributing factor in this regard. METHODS: Using data from a multi-center case-control study in Iran, base line testosterone, sex hormone binding globulin (SHBG), estradiol, and albumin levels as well as thorough demographic and medical characteristics of 194 newly diagnosed prostate cancer patients were determined. There were 317 ethnicity matched men with no cancer as controls as well. Data was analyzed for hormones of interest in DM patients regarding their cancer status. RESULTS: Of 511 enrolled patients, twenty-one cases and 63 controls were diagnosed as DM. Patients with DM were significantly less likely to have PC (OR: 0.44, P = 0.003). Time since DM diagnosis was also inversely correlated with the risk of cancer (P trend < 0.0001). Control patients had significantly higher testosterone, estradiol, and testosterone/SHBG ratio (P < 0.05). As time since DM diagnosis increased by quartiles, testosterone significantly increased (P trend < 0.05). The risk of PC also significantly declined (P trend < 0.0001) following an initial remarkable increase early after DM diagnosis. After including the hormones in the logistic regression model, there was a weak, yet significant inverse association of testosterone/SHBG and DM duration with the risk of PC. CONCLUSIONS: Based on our results DM duration is inversely correlated with the risk of prostate cancer. Our results do not support the hypothesis that sex hormones, including testosterone, play a major role in the protective effect of DM against PC. PMID- 19676083 TI - PSA surge/flare-up in patients with castration-refractory prostate cancer during the initial phase of chemotherapy. AB - BACKGROUND: Docetaxel-based chemotherapy has shown great promise for the treatment of CRPC and is considered the current standard of care. PSA is mainly used as marker to monitor the treatment response. Several articles were published reporting an initial PSA surge/flare-up after starting chemotherapy. The cause and the impact of this phenomenon are discussed controversially. The intention of this review is to define the significance of initial PSA surge/flare-up and to increase awareness to this phenomenon in the urological community. MATERIALS AND METHODS: A comprehensive literature search was performed in different data bases using various key words. Relevant articles and references between 1999 and 2009 were reviewed and analyzed for data on the association between chemotherapy and initial PSA surge/flare. RESULTS: The incidence of a PSA surge/flare-up ranges according to the reported studies between 7.6% and 13.6%. A PSA surge/flare-up was reported up to 404% from baseline PSA level followed by PSA response. The median duration of a PSA surge/flare-up is 2-3 weeks and can last up to 6-8 weeks. However, the occurrence of a PSA surge/flare-up did not impact outcome and survival negatively compared to patients with an immediate PSA response. CONCLUSIONS: A considerable portion of CRPC patients experience an initial PSA surge/flare-up under systemic chemotherapy. The definitions used for PSA surge/flare-up differ slightly in the literature. This issue needs to be solved since it might impact defining treatment response. As a PSA surge/flare-up did not impact outcome and survival negatively, chemotherapy should be continued according to the literature addressing specifically the phenomenon of a PSA surge/flare-up for a minimum of 8 weeks or 3 rounds of a 3-weekly cycle chemotherapy regimen before further decisions are made about efficacy. However, Scher et al. recommended a 12-week period drug exposure based on their results on PSA progression-free survival and overall survival. This dilemma needs to be addressed in further data analysis in order to establish a general rule regarding when to stop chemotherapy. Physicians should be aware of this effect to avoid inadequate early discontinuation of chemotherapy. The underlying mechanisms of a PSA surge/flare-up are still elusive and need further clarification. PMID- 19676084 TI - CE immunoassay with enhanced chemiluminescence detection of erythropoietin using silica dioxide nanoparticles as pseudostationary phase. AB - The measurement of serum erythropoietin (EPO) is important for the detection of recombinant human EPO misuse in sports. In this paper, we have developed a sensitive and rapid CE immunoassay with enhanced chemiluminescence detection of EPO, in which silica dioxide nanoparticles (SiO2 NPs) were used as the pseudostationary phase to improve the separation efficiency of analytes. By adopting SiO2 NPs in the CE immunoassay, the separation can be successfully performed in neutral running buffer solution. However, running buffer of extreme pH was still necessary for the conventional CZE mode. Neutral phosphate buffer (pH 7.4) containing SiO2 NPs and poly(ethylene oxide) (PEO) was chosen as running buffer. The influences of SiO2 NPs, PEO and buffer concentration on the separation efficiency and EPO detection were investigated. EPO-horse radish peroxidase (HRP) and immunocomplex were baseline separated in a 10 mM phosphate buffer (pH 7.4) consisting of 0.08% PEO and 0.08% SiO2 NPs. The linear range for EPO was 1.8-158.0 ng/mL and the detection limit was 0.9 ng/mL. The assay was successfully applied for the quantification of EPO in human sera and the results correlated well with those obtained using chemiluminescence immunoassay kits, thus demonstrating that the present method was a potential powerful tool for EPO misuse detection and clinical diagnosis. PMID- 19676085 TI - The effects of microstructures on a dielectrophoretic chip for trapping particles. AB - A dielectrophoretic (DEP) chip with an SU-8 microcavity array for trapping single particle/cell is designed, fabricated, and quantitatively examined by simulations and experiments. The particles can be easily trapped in or pulled out of the microcavity based on negative or positive DEP force, respectively. The nonuniform electric field is formed in relation to the configuration of the microcavity array, i.e. its diameter and spacing, as described in the simulation results. In order to investigate the effects of the microcavity, two maximal flow rates for trapping particles in the microcavity and washing them away from the microcavity under different DEP voltages are determined by experiments. As the experimental results show, the extrastationary effects, provided as a particle is trapped in the microcavity, mean that the trapped particle can sustain a low flow rate even without applying DEP voltage. Consequently, this DEP chip is suitable for long term monitoring of trapped cells by supplying the subsisting buffer at a low flow rate without the damage or heating effect caused by DEP voltage. PMID- 19676086 TI - Complete sequencing and oxidative modification of manganese superoxide dismutase in medulloblastoma cells. AB - Manganese superoxide dismutase (Mn-SOD or SOD2) is a key antioxidant enzyme and was assigned several roles in tumor biology. Working on medulloblastoma cell line DAOY, we identified two spots as Mn-SODs. Because of the proposed pivotal role of this enzyme in oncobiology, we decided to completely sequence the proteins and to determine PTMs. Proteins extracted from DAOY cells were run on 2-DE, multienzyme digestions were carried out and peptides were analyzed by MALDI-TOF/TOF, Qq-TOF and the ion trap using both the CID and ETD principles. Both protein expression forms were completely sequenced and revealed identical protein sequences. Histidines His30 and His31 were oxidized in one protein, whereas tryptophan oxidation (Trp-186) was observed in both. Histidine oxidation was not only indicated by the mass shift of the peptide but also by specific spectra of 2-oxo histidine and a previously described intermediate (His+14). Complete sequencing of the two Mn-SOD expression forms unambiguously characterizes this enzyme from a tumor cell line providing evidence that can be used for generation of antibodies and allowing conformational studies. The findings of different PTMs in the same gel represent Mn-SOD oxidative states, while oxidative modification of His30 and 31 may even reflect decreased Mn-SOD activity. PMID- 19676087 TI - An efficient method for the extraction of chloroplast proteins compatible for 2 DE and MS analysis. AB - Comparative proteomic analysis of chloroplast by 2-DE has received significant attention in recent years. However, the complication of membrane systems in chloroplast made it challenging to elucidate entire chloroplast proteome by 2-DE. Here, we developed an efficient method for extracting chloroplast proteins, and produced excellent 2-DE profiles from both Arabidopsis thaliana and Salicornia europaea. Comparison of this method with another two protocols for the extraction of A. thaliana chloroplast proteins showed that our method obtained higher protein yields and produced more protein spots on both pH 3-10 and 4-7 2-DE gels. Moreover, this method recovered more proteins in the basic and high M(r) regions, thereby offering the best extraction of chloroplast proteins. Identification of 15 specific chloroplast-targeted proteins on our gels by MALDI-TOF MS revealed that this method was compatible with MS, and recovered more chloroplast membrane proteins than the commonly used methods. This protocol is expected to have a wide application in future chloroplast proteomic analysis. PMID- 19676088 TI - Differentiation of Staphylococcus aureus strains by CE, zeta potential and coagulase gene polymorphism. AB - Staphylococcus aureus is a common cause of infection in both hospitals and the community, and it is becoming increasingly virulent and resistant to antibiotics. Possibilities of fast, sensitive and cheap determination of these pathogenic bacteria are extremely important in antimicrobial therapy. In the present study, CE with chemically modified capillary and zeta potential measurements were used for differentiation of three different clinical strains of S. aureus. The data presented in this contribution suggested that electrophoretic behavior and the values of zeta potential should be very useful in distinguishing between closely related strains, which exhibited coagulase gene/protein polymorphism. Understanding the differences between S. aureus strains could help to improve our knowledge about S. aureus pathogenecity and to monitor for and respond to emergence of more virulent strains. PMID- 19676089 TI - Determination of DNA methylation by COBRA: a comparative study of CGE with LIF detection and conventional gel electrophoresis. AB - DNA methylation as an epigenetic modification of the human genome is under emphatic investigation. Several studies have demonstrated a role of DNA methylation in oncogenesis. In conjunction with histone modifications, DNA methylation may cause the formation of heterochromatin and thus mediate the inactivation of gene transcription. It is important to develop methods that allow for an accurate quantification of the amount of DNA methylation in particular DNA regions, to gain information concerning the threshold of methylation levels necessary for gene inactivation. In this article, a CGE method with on-column LIF detection using SYBR Green is compared with a conventional slab-gel electrophoresis. We thus investigate the validity to analyze DNA methylation in the samples of a combined bisulfite restriction analysis. It is demonstrated that CGE is superior to gel electrophoresis in means of linearity, precision, accuracy, automatization (high throughput), and sample consumption. However, gel electrophoresis is easier to perform (simple devices, no PC usage), and the running costs are comparatively low. A further advantage of CGE is the sparse use of toxic compounds (MeOH and SYBR Green), whereas gel electrophoresis is performed in polyacrylamide gels with ethidium bromide staining. PMID- 19676090 TI - The consequences of sample pooling in proteomics: an empirical study. AB - Pooling of samples in proteomics experiments might help overcome resource constraints when many individuals are analysed. The measured biological variation should be reduced giving increased power to detect treatment differences. Pooling has been advocated in microarray work but there are few tests of its potential in proteomics. In this study, we examine three issues on which the success of the pooling approach might hinge and provide evidence that: (i) the protein expression in a pool matches the mean expression of the individuals making up the pool for the majority of proteins, although for some proteins the pool expression is different; (ii) the biological variance between pools is reduced compared with that between individuals, as predicted in theory, but this reduction is not as large as expected. A practical consequence of this is that power could be reduced; (iii) proteins detectable in individual samples are usually but not always visible when samples are pooled. We conclude that pooling of samples in proteomics work is a valid and potentially valuable procedure but consideration should be given to these issues in experimental design. PMID- 19676091 TI - Proteomic analysis of date palm (Phoenix dactylifera L.) responses to endophytic colonization by entomopathogenic fungi. AB - The entomopathogenic fungi Beauveria bassiana, Lecanicillium dimorphum and L. cf. psalliotae can survive and colonize living palm tissue as endophytes. The molecular interaction between these biocontrol agent fungi and the date palm Phoenix dactylifera L. was investigated using proteomic techniques. Field date palms inoculated with these fungi were analyzed 15 and 30 days after inoculation in two independent bioassays. In vitro date palms were also inoculated with B. bassiana or L. cf. psalliotae. Qualitative and quantitative differences in protein accumulation between controls (not inoculated) and inoculated palms were found using 2-DE analysis, and some of these responsive proteins could be identified using MALDI/TOF-TOF. Proteins involved in plant defence or stress response were induced in P. dactylifera leaves as a response to endophytic colonization by entomopathogenic fungi in field date palms. Proteins related with photosynthesis and energy metabolism were also affected by entomopathogenic fungi colonization. A myosin heavy chain-like protein was accumulated in in vitro palms inoculated with these fungi. This suggests that endophytic colonization by these entomopathogenic fungi modulates plant defence responses and energy metabolism in field date palms and possibly modulates the expression of cell division-related proteins in in vitro palms at proteomic level. PMID- 19676092 TI - Establishing the mitochondrial DNA D-loop structure of Columba livia. AB - The complete mitochondrial DNA D-loop structure of pigeon (Columba livia) was established in this study. A strategy of amplifying three partial fragments of the D-loop and then combing the three fragments to cover the full length of the D loop was adopted. Ten samples from pigeons were collected and were successfully amplified and sequenced. Repetitive sequences of a VNTR and an STR were both observed at the 3'-end of D-loop region. DNA sequence data revealed polymorphic sequences including indels, SNP, VNTR and STR within the D-loop. The size of the D-loop ranged from 1310 to 1327 bp from the initiation site of D-loop to the site immediately upstream of the repeat sequences depending upon the number of insertions or deletions. Each sample could be distinguished based on four genotyping procedures; being indels, SNPs, VNTRs and STRs. The polymorphic nature of the D-loop can be a valuable method for maternal identification and genetic linkage of pigeon in particular forensic science investigations. PMID- 19676093 TI - Tissue culture media supplemented with 10% fetal calf serum contains a castrate level of testosterone. AB - BACKGROUND: Human prostate cancer cells are routinely maintained in media supplemented with 10% Fetal Calf Serum (FCS) to provide androgen. In the present study, total and free testosterone levels in 10%FCS supplemented tissue culture media were determined and compared to levels in intact and castrated human males. Dextran-coated charcoal stripped FCS (i.e., DC-FCS) is often used instead of FCS to minimize the level of androgen provided in 10% serum supplemented media. Therefore, total and free testosterone levels in 10%DC-FCS containing media were likewise determined. METHODS: Total testosterone, free testosterone, and total dihydrotestosterone (DHT) were determined on RPMI-1640 media supplemented with either 10%FCS or 10%DC-FCS by ELISA assays before and after exposure to LNCaP human prostate cancer cells in culture. The growth and PSA secretion by these cells was also determined. RESULTS: Ten percentage FCS supplemented media contains a castrate level of testosterone. However, even with this castrate starting level of testosterone, LNCaP cells concentrate and metabolize the testosterone to produce a physiologic (i.e., 10 nM) level of intracellular DHT which optimally stimulates the growth of these cells in vitro. CONCLUSIONS: The present studies document that prostate cancer cells auto-regulate their androgen metabolism so that an optimal level of DHT for growth is maintained during both up and down fluctuations in the supply of testosterone. These results have significant implications for whether exogenous androgen should be added to the 10%FCS supplemented media to grow prostate cancer cells from intact versus castrated patients. PMID- 19676094 TI - The expression of androgen-responsive genes is up-regulated in the epithelia of benign prostatic hyperplasia. AB - BACKGROUND: Benign prostatic hyperplasia (BPH) is one of the most common diseases among aging men in the United States. In addition to aging, the presence of androgens is another major risk factor in BPH development. However, whether androgen signaling is altered in BPH remains unclear. To determine androgen signaling in BPH, we characterized the expression of four different androgen responsive genes, Eaf2/U19, ELL2, FKBP5, and PSA, in BPH and adjacent normal glandular epithelial cells. METHODS: A set of 17 BPH specimens were resected from patients over 60 years of age with clinical symptoms of BPH. Laser-capture microdissection (LCM) was used to isolate glandular epithelial cells from BPH areas and adjacent normal areas, separately. LCM isolated cells from individual specimens were lysed and RNA isolation, reverse transcription, and real-time PCR were performed using CellsDirect One-Step qRT-PCR Kit (Invitrogen, Carlsbad, CA). RESULTS: All of the assayed genes displayed increased expression, from approximately 2- to approximately 6-fold, in BPH as compared to the adjacent normal epithelial cells. We also generated a composite androgen response index based on the expression levels of the four genes, which provides a reliable readout for overall androgen action. Our study showed that the composite androgen response index in BPH is approximately 4-fold as compared to that in the adjacent normal tissues. CONCLUSIONS: Androgen signaling is significantly elevated in BPH relative to the adjacent normal prostate. Understanding the mechanisms causing elevated androgen signaling may lead to novel approaches for prevention and/or treatment of BPH. PMID- 19676095 TI - Adiponectin increases motility of human prostate cancer cells via adipoR, p38, AMPK, and NF-kappaB pathways. AB - BACKGROUND: Prostate cancer is the most commonly diagnosed malignancy in men. Prostate cancer shows a predilection for metastasis to the bone. Adiponectin is a protein hormone secreted predominantly by differentiated adipocytes and involved in energy homeostasis. The aim of this study was to investigate whether adiponectin is associated with migration of prostate cancer cells. METHOD: Cancer cells migration activity was examined using the Transwell assay. The p38 and AMPK phosphorylation was examined by using Western blot method. The cell surface expression of integrins was examined by using flow cytometry. The qPCR was used to examine the mRNA expression of integrin. A transient transfection protocol was used to examine NF-kappaB activity. RESULTS: We found that adiponectin increased the migration and the expression of alpha5beta1 integrin of human prostate cancer cells. Adiponectin-mediated migration and integrins expression was attenuated by p38 inhibitor (SB203580), p38 mutant, AMPK siRNA, AMPK inhibitor (araA and compound C). Activations of p38, AMPK and NF-kappaB pathways after adiponectin treatment was demonstrated, and adiponectin-induced expression of integrins and migration activity was inhibited by the specific inhibitor and mutant of p38, AMPK, and NF-kappaB cascades. CONCLUSIONS: This study showed for the first time that the adiponectin mediates migration of human prostate cancer cells. One of the mechanisms underlying adiponectin directed migration was transcriptional up regulation of alpha5beta1 integrin and activation of AdipoR1 receptor, p38, AMPK, and NF-kappaB pathways. PMID- 19676096 TI - Reduced 3-O-methyl-dopa levels in OCD patients and their unaffected parents is associated with the low activity M158 COMT allele. AB - The catechol-O-methyltransferase (COMT) gene is considered as a candidate gene in obsessive-compulsive disorder (OCD). Specifically, the COMT low-activity M158 allele has been suggested to be associated with OCD. However, there is no study reporting that COMT activity is decreased in OCD patients and that the decrease is mediated by the V158M polymorphism. Therefore, the purpose of our study was to assess COMT activity in OCD by measuring plasma levels of 3-O-methyl-dopa (3 OMD), which result from the methylation of levodopa by COMT, and to investigate the relationship between 3-OMD levels and the V158M polymorphism. We also examined whether 3-OMD levels represented an endophenotype, associated with the genetic liability to OCD, by assessing unaffected relatives of OCD patients. We assessed plasma 3-OMD levels in a sample of drug-free OCD probands (n = 34) and their unaffected parents (n = 63), and compared them with controls (n = 85). The COMT V158M polymorphism was genotyped in all participants. Lower plasma 3-OMD levels were found in OCD probands and their unaffected parents compared to controls. The COMT M158 allele was associated with reduced plasma 3-OMD levels in a co-dominant manner, both in OCD probands and their relatives, but not in controls. Our results suggest that COMT activity could act as a limiting factor for the production of 3-OMD in OCD patients and in their relatives. These findings further support a role of COMT in the susceptibility to OCD and provide evidence that 3-OMD levels could represent an endophenotype in OCD. PMID- 19676098 TI - Tandem mass spectrometry of synthetic polymers. AB - The detailed characterization of macromolecules plays an important role for synthetic chemists to define and specify the structure and properties of the successfully synthesized polymers. The search for new characterization techniques for polymers is essential for the continuation of the development of improved synthesis methods. The application of tandem mass spectrometry for the detailed characterization of synthetic polymers using the soft ionization techniques matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and electrospray ionization mass spectrometry (ESI-MS), which became the basic tools in proteomics, has greatly been increased in recent years and is summarized in this perspective. Examples of a variety of homopolymers, such as poly(methyl methacrylate), poly(ethylene glycol), as well as copolymers, e.g. copolyesters, are given. The advanced mass spectrometric techniques described in this review will presumably become one of the basic tools in polymer chemistry in the near future. PMID- 19676097 TI - FKBP5 polymorphisms and antidepressant response in geriatric depression. AB - Genetic variation at the FKBP5 locus has been reported to affect clinical outcomes in patients treated with antidepressant medications in several studies. However, other reports have not confirmed this association. FKBP5 may regulate the sensitivity of the hypothalamic-pituitary-adrenal axis. We tested two FKBP5 single nucleotide polymorphisms (rs1360780 and rs3800373) in a sample of 246 geriatric patients treated for 8 weeks in a double-blind randomized comparison trial of paroxetine and mirtazapine. These two polymorphisms had previously been reported to predict efficacy in depressed patients treated with selective serotonin reuptake inhibitors such as paroxetine, and those treated with mirtazapine, an agent with both serotonergic and noradrenergic actions. However, we found no significant associations between these FKBP5 genetic variants and clinical outcomes. Neither mean Hamilton Depression Rating Scale scores nor time to remission or response were predicted by FKBP5 genetic variation. These results suggest that FKBP5 is unlikely to play a major role in determining antidepressant treatment outcomes in geriatric patients. PMID- 19676099 TI - Gas phase behavior of radical cations of perfluoroalkyl-1,2,4-triazines: an experimental and theoretical study. AB - Electron ionization mass spectrometry and low-energy collision-induced decomposition reactions occurring in a tridimensional ion trap, together with density functional theory (DFT) calculations on neutrals, even- and odd-electron cations, have been used to study the gas-phase ion chemistry of a series of perfluoroalkyl-1,2,4-triazines. Loss of oxygen, due to thermal degradation occurring before ionization, likely involving the hydroxylamino group, has been observed. Compounds having a carbonyl group at position 6 of the triazine ring fragment in the source by elimination of NO followed by HF or CO. The decomposition pathways occurring due to CID experiments have shown interesting features depending on the nature and structure of precursor ions. Most of them involve elimination of endocyclic atoms, thereby producing contraction of the original six-membered ring or formation of acyclic structures. DFT (B3LYP/6 31G(d,p)) calculations have been used for evaluating structure, stability and properties of neutral and ionic species involved in gas-phase processes. In particular, it has been calculated that in the molecular ion the unpaired electron is mainly located on the exocyclic nitrogen, while the positive charge is on the C(6) carbon atom. PMID- 19676100 TI - Inactivation of SNF5 cooperates with p53 loss to accelerate tumor formation in Snf5(+/-);p53(+/-) mice. AB - Malignant rhabdoid tumors (MRTs) are poorly differentiated pediatric cancers that arise in various anatomical locations and have a very poor outcome. The large majority of these malignancies are caused by loss of function of the SNF5/INI1 component of the SWI/SNF chromatin remodeling complex. However, the mechanism of tumor development associated with SNF5 loss remains unclear. Multiple studies have demonstrated a role for SNF5 in the regulation of cyclin D1, p16(INK4A), and pRb(f) activities suggesting it functions through the SWI/SNF complex to affect transcription of genes involved in cell cycle control. Previous studies in genetically engineered mouse models (GEMM) have shown that loss of SNF5 on a p53 null background significantly accelerates tumor development. Here, we use established GEMM to further define the relationship between the SNF5 and p53 tumor suppressor pathways. Combined haploinsufficiency of p53 and Snf5 leads to decreased latency for MRTs arising in alternate anatomical locations but not for the standard facial MRTs. We also observed acceleration in the appearance of T cell lymphomas in the p53(+/-);Snf5(+/-) mice. Our studies suggest that loss of SNF5 activity does not bestow a selective advantage on the p53 spectrum of tumors in the p53(+/-);Snf5(+/-) mice. However, reduced p53 expression specifically accelerated the growth of a subset of MRTs in these mice. PMID- 19676103 TI - Real-time tracking of vocal fold injections with optical coherence tomography. AB - OBJECTIVES/HYPOTHESIS: Optical coherence tomography (OCT) is a promising new imaging modality that can help discern the layered microstructure of vocal folds. In the future, subepithelial injections of implants will improve vocal fold pliability where there is stiffness of phonatory mucosa. Using OCT both to delineate the depth of subepithelial injections real-time and to serially image the implant over time would be valuable, and has not been demonstrated previously. STUDY DESIGN: Ex vivo study using excised calf larynges and survival study using canines in vivo. METHODS: An investigation was done employing real time OCT imaging of subepithelial injection pulses into phonatory mucosa of four calf larynges ex vivo to track the presence of subepithelial implants in phonatory mucosa in a survival study using an in vivo canine model. RESULTS: OCT readily identified polyethylene glycol (PEG)-based hydrogel in the subepithelial plane in both the ex vivo calf study and the in vivo canine study. Ex vivo calf images correlated with histological specimens obtained immediately postinjection. Images obtained in this study provide confirmation of the hydrogel injection depth in real time, and allow for the implant to be tracked during a canine survival study. CONCLUSIONS: OCT can confirm subepithelial placement of hydrogel implant in the vocal fold with sufficient resolution to provide instantaneous feedback of an injection pulse. Survival studies in an in vivo canine model indicate that OCT can potentially be helpful in monitoring rheologically appropriate implants within the superficial lamina propria. PMID- 19676102 TI - Common origin of pure and interrupted repeat expansions in spinocerebellar ataxia type 2 (SCA2). AB - The spinocerebellar ataxia type 2 (SCA2) is an autosomal dominant neurodegenerative disease characterized by gait and limb ataxia. This disease is caused by the expansion of a (CAG)(n) located in the ATXN2, that encodes a polyglutamine tract of more than 34 repeats. Lately, alleles with 32-33 CAGs have been associated to late-onset disease cases. Repeat interruptions by CAA triplets are common in normal alleles, while expanded alleles usually contain a pure repeat tract. To investigate the mutational origin and the instability associated to the ATXN2 repeat, we performed an extensive haplotype study and sequencing of the CAG/CAA repeat, in a cohort of families of different geographic origins and phenotypes. Our results showed (1) CAA interruptions also in expanded ATXN2 alleles; (2) that pathological CAA interrupted alleles shared an ancestral haplotype with pure expanded alleles; and (3) higher genetic diversity in European SCA2 families, suggesting an older European ancestry of SCA2. In conclusion, we found instability towards expansion in interrupted ATXN2 alleles and a shared ancestral ATXN2 haplotype for pure and interrupted expanded alleles; this finding has strong implications in mutation diagnosis and counseling. Our results indicate that interrupted alleles, below the pathological threshold, may be a reservoir of mutable alleles, prone to expansion in subsequent generations, leading to full disease mutation. PMID- 19676101 TI - Effect of dopamine transporter gene (SLC6A3) variation on dorsal anterior cingulate function in attention-deficit/hyperactivity disorder. AB - Although attention-deficit/hyperactivity disorder (ADHD) is associated both with brain alterations in attention and executive function (EF) circuitry and with genetic variations within the dopamine system (including the dopamine transporter gene [SLC6A3]), few studies have directly investigated how genetic variations are linked to brain alterations. We sought to examine how a polymorphism in the 3' untranslated region (UTR) of SLC6A3, associated with ADHD in meta-analysis, might contribute to variation in dorsal anterior cingulate cortex (dACC) function in subjects with ADHD. We collected fMRI scans of 42 individuals with ADHD, all of European descent and over the age of 17, while they performed the multi-source interference task (MSIT), a cognitive task shown to activate dACC. SLC6A3 3' UTR variable number tandem repeat (VNTR) polymorphisms were genotyped and brain activity was compared for groups based on allele status. ADHD individuals homozygous for the 10R allele showed significant hypoactivation in the left dACC compared to 9R-carriers. Exploratory analysis also showed trends toward hypoactivation in the 10R homozygotes in left cerebellar vermis and right lateral prefrontal cortex. Further breakdown of genotype groups showed similar activation in individuals heterozygous and homozygous for the 9R allele. Alterations in activation of attention and EF networks found previously to be involved in ADHD are likely influenced by SLC6A3 genotype. This genotype may contribute to heterogeneity of brain alterations found within ADHD samples. PMID- 19676104 TI - Betuletol 3-methyl ether induces G(2)-M phase arrest and activates the sphingomyelin and MAPK pathways in human leukemia cells. AB - Betuletol 3-methyl ether (BME) is a natural phenylbenzo-gamma-pyrone that inhibits cell proliferation in human tumor cell lines and induces apoptotic cell death in HL-60 cells. Here we show that BME displays strong cytotoxic properties in several human leukemia cell lines (U937, K-562, THP-1, Jurkat, and Molt-3) and in cells that over-express two anti-apoptotic proteins, namely Bcl-2 and Bcl x(L). BME arrested HL-60 cells at G(2)-M phase of the cell cycle, which was associated with the accumulation of cyclin B1 and p21(Cip1). Fluorescence microscopy experiments suggest that BME blocked the cell cycle in mitosis. The in vivo tubulin polymerization assay shows that BME inhibits tubulin polymerization and causes similar changes of cellular microtubule network as colchicine. Our results demonstrate that BME-induced cell death is (i) triggered in human myeloid leukemia cell that over-express Bcl-2 and Bcl-x(L), and (ii) associated with loss of inner mitochondrial membrane potential (DeltaPsim) and an increase in reactive oxygen species (ROS). Although ROS increased in response to BME, this did not seem to play a pivotal role in the apoptotic process since the anti-oxidant trolox was unable to provide cell protection. The treatment of HL-60 cells with BME induces the activation of mitogen-activated protein kinases (MAPKs) such as c Jun N-terminal kinases, p38 mitogen-activated protein kinases and extracellular signal-regulated kinases (ERK)1/2 and stimulates the acid sphingomyelinase with concomitant ceramide generation. The findings of this study suggest that BME could be useful in the development of novel anticancer agents. PMID- 19676105 TI - Curcumin-induced apoptosis in ovarian carcinoma cells is p53-independent and involves p38 mitogen-activated protein kinase activation and downregulation of Bcl-2 and survivin expression and Akt signaling. AB - New cytotoxic agents are urgently needed for the treatment of advanced ovarian cancer because of the poor long-term response of this disease to conventional chemotherapy. Curcumin, obtained from the rhizome of Curcuma longa, has potent anticancer activity; however, the mechanism of curcumin-induced cytotoxicity in ovarian cancer cells remains a mystery. In this study we show that curcumin exhibited time- and dose-dependent cytotoxicity against monolayer cultures of ovarian carcinoma cell lines with differing p53 status (wild-type p53: HEY, OVCA429; mutant p53: OCC1; null p53: SKOV3). In addition, p53 knockdown or p53 inhibition did not diminish curcumin killing of HEY cells, confirming p53 independent cytotoxicity. Curcumin also killed OVCA429, and SKOV3 cells grown as multicellular spheroids. Nuclear condensation and fragmentation, as well as DNA fragmentation and poly (ADP-ribose) polymerase-1 cleavage in curcumin-treated HEY cells, indicated cell death by apoptosis. Procaspase-3, procaspase-8, and procaspase-9 cleavage, in addition to cytochrome c release and Bid cleavage into truncated Bid, revealed that curcumin activated both the extrinsic and intrinsic pathways of apoptosis. Bax expression was unchanged but Bcl-2, survivin, phosphorylated Akt (on serine 473), and total Akt were downregulated in curcumin treated HEY cells. Curcumin also activated p38 mitogen-activated protein kinase (MAPK) without altering extracellular signal-regulated kinase 1/2 activity. We conclude that p53-independent curcumin-induced apoptosis in ovarian carcinoma cells involves p38 MAPK activation, ablation of prosurvival Akt signaling, and reduced expression of the antiapoptotic proteins Bcl-2 and survivin. These data provide a mechanistic rationale for the potential use of curcumin in the treatment of ovarian cancer. PMID- 19676106 TI - Gastric cancer risk predisposition and prognostic significance of vascular endothelial growth factor (VEGF) gene polymorphisms--a case-control study in an Omani population. AB - Vascular endothelial growth factor (VEGF) plays a central role in angiogenesis, tumor growth, and metastasis. We investigated the associations between VEGF gene polymorphisms and gastric cancer (GC) risk predisposition and prognostic characteristics in an Omani population, an ethnic group which has not been studied previously. We analyzed three VEGF polymorphisms (+405 G/C, -460 T/C, and +936 C/T) by the extraction of genomic DNA from peripheral blood of 130 GC patients and 130 control subjects followed by VEGF genotyping using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis. There were no significant associations between the VEGF polymorphisms and GC risk. There were significant correlations between the +405 C/C genotype and both poor tumor differentiation (P = 0.007) and lymph node metastasis (P = 0.03) and between the -460 T/T genotype and poor tumor differentiation (P = 0.03) with a statistical trend toward lymph node involvement (P = 0.05). VEGF gene polymorphisms had no significant effects on survival, but the VEGF +405 G/G genotype had a statistical trend toward lower survival rate with a hazard ratio of 1.6 [95% CI, 0.9-2.9] compared with the VEGF +405 CC/GC combined genotype (P = 0.049). Multivariate analysis showed that disease stage at diagnosis and the +405 G/G genotype were independent variables of adverse prognostic significance. There were no associations between the six common haplotypes identified and both GC risk predisposition and survival. The current study suggests that VEGF polymorphisms have no role in GC risk predisposition, but may have prognostic significance in GC patients. PMID- 19676107 TI - Inhibited temperament and parent emotional availability differentially predict young children's cortisol responses to novel social and nonsocial events. AB - Preschool-aged children (n = 274) were examined in the laboratory to assess behavioral and cortisol responses to nonsocial and social threat. Parents also responded to scales on the Children's Behavior Questionnaire reflecting exuberant approach to novel/risky activities (reversed scored) and shyness. Multi-method measures of Nonsocial and Social Inhibition were computed. Parents and children were observed engaging in a series of interactive tasks and the Emotional Availability scales were scored for parental sensitivity, nonintrusiveness, nonhostility, and structuring. These scores were factored to yield one measure of Parenting Quality. Analyses revealed that Nonsocial and Social Inhibition could be distinguished and that associations with cortisol response were stressor specific. Moderation analyses revealed that parenting quality buffered cortisol elevations for extremely socially, but not nonsocially inhibited children. These findings are consistent with evidence that sensitive, supportive parenting is an important buffer of the HPA axis response to threat in infants and toddlers, and extends this finding to the preschool period. PMID- 19676108 TI - Mcl-1 and Bcl-xL cooperatively maintain integrity of hepatocytes in developing and adult murine liver. AB - Anti-apoptotic members of the Bcl-2 family, including Bcl-2, Bcl-xL, Mcl-1, Bcl-w and Bfl-1, inhibit the mitochondrial pathway of apoptosis. Bcl-xL and Mcl-1 are constitutively expressed in the liver. Although previous research established Bcl xL as a critical apoptosis antagonist in differentiated hepatocytes, the significance of Mcl-1 in the liver, especially in conjunction with Bcl-xL, has not been clear. To examine this question, we generated hepatocyte-specific Mcl-1 deficient mice by crossing mcl-1(flox/flox) mice and AlbCre mice and further crossed them with bcl-x(flox/flox) mice, giving Mcl-1/Bcl-xL-deficient mice. The mcl-1(flox/flox) AlbCre mice showed spontaneous apoptosis of hepatocytes after birth, as evidenced by elevated levels of serum alanine aminotransferase (ALT) and caspase-3/7 activity and an increased number of terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate nick-end labeling (TUNEL) positive cells in the liver; these phenotypes were very close to those previously found in hepatocyte-specific Bcl-xL-deficient mice. Although mcl-1(flox/+) AlbCre mice did not display apoptosis, their susceptibility to Fas-mediated liver injury significantly increased. Further crossing of Mcl-1 mice with Bcl-xL mice showed that bcl-x(flox/+) mcl-1(flox/+) AlbCre mice also showed spontaneous hepatocyte apoptosis similar to Bcl-xL-deficient or Mcl-1-deficient mice. In contrast, bcl x(flox/flox) mcl-1(flox/+) AlbCre, bcl-x(flox/+) mcl-1(flox/flox) AlbCre, and bcl x(flox/flox) mcl-1(flox/flox) AlbCre mice displayed a decreased number of hepatocytes and a reduced volume of the liver on day 18.5 of embryogenesis and rapidly died within 1 day after birth, developing hepatic failure evidenced by increased levels of blood ammonia and bilirubin. CONCLUSION: Mcl-1 is critical for blocking apoptosis in adult liver and, in the absence of Bcl-xL, is essential for normal liver development. Mcl-1 and Bcl-xL are two major anti-apoptotic Bcl-2 family proteins expressed in the liver and cooperatively control hepatic integrity during liver development and in adult liver homeostasis in a gene dose dependent manner. PMID- 19676109 TI - An evaluation of lower-body functional limitations among long-term survivors of 11 different types of cancers. AB - BACKGROUND: The authors examined potential reasons (sociodemographics, psychologic distress, health behavior, chronic health conditions, access to medical care) for increased prevalence of lower-body functional limitations among long-term (> or =5 years) cancer survivors. METHODS: The authors used National Health Interview Survey data from 2005 through 2007, and defined lower-body functional limitation as reporting difficulty/inability to perform at least 1 of 5 activities (walking approximately one-quarter of a mile; walking up and down 10 steps without rest; standing for 2 hours; stooping, crouching, or kneeling; and lifting 10 lbs). Increased prevalence of lower-body functional limitations was compared between long-term survivors of each of 11 cancer types reported by > or =50 respondents (n = 2143) and persons without cancer history (controls; n = 72,618). RESULTS: Among cancer survivors, 57.0% had a lower-body functional limitation versus 26.6% of controls. The unadjusted prevalence of lower-body functional limitations varied by cancer type, ranging from 44.9% (lymphoma survivors) to 88.8% (lung cancer survivors). Long-term lung (odds ratio [OR], 7.91), uterine (OR, 2.41), thyroid (OR, 2.27), cervical (OR, 1.76), ovarian (OR, 1.75), and breast (OR, 1.35) cancer survivors had increased odds of reporting a lower-body functional limitation than controls after adjusting for sociodemographic factors (all P < .05). Differences in the prevalence of arthritis and lower-back pain and in access to medical care explained differences in lower-body functional limitation prevalence between controls and long-term breast, cervical, ovarian, and uterine cancer survivors. Long-term bladder, colorectal, lymphoma, melanoma, and prostate cancer survivors were equally likely to report a lower-body functional limitation as controls. CONCLUSIONS: Treatment of arthritis and lower-back pain and increasing access to medical care might help reduce the risk of lower-body functional limitations and improve quality of life among specific long-term cancer survivors. PMID- 19676110 TI - Anger, aggression, and self-harm in PTSD and complex PTSD. AB - This study examined the contribution of complex posttraumatic stress disorder (PTSD) diagnosis and symptomatology to the difficulties of anger, aggression, and self-harm in a Northern Ireland clinical community sample. A "current complex PTSD" (CCPTSD) group (n=11) was compared with a "current PTSD" group (n=31) on self-report measures of these variables. The CCPTSD group demonstrated significantly higher levels of physical aggression and self-harm than the PTSD group. The complex PTSD symptom of 'alterations in self-perception' was a significant predictor of aggression and history of self-harm, suggesting the potential role of posttraumatic shame and self-loathing in PTSD theoretical models of these destructive behaviors. Social desirability was a notable confounding influence in the assessment of anger, aggression, and self-harm in traumatised individuals. PMID- 19676112 TI - Varying patterns of coexistence of two mouse lemur species (Microcebus ravelobensis and M. murinus) in a heterogeneous landscape. AB - The coexistence of closely related species is not easily understood on the basis of ecological theories. This study investigates the extent of coexistence of two congeneric species of Microcebus murinus (MUR) and M. ravelobensis (RAV) in northwestern Madagascar. Their presence and local relative population densities were determined by capturing and nocturnal transect counts and compared at 22 study sites in the Ankarafantsika National Park. All sites were characterized with regard to their altitude, access to surface water, and 19 structural vegetation characteristics. RAV and MUR were not equally distributed over this regional scale. RAV occurred in more sites and at higher maximum densities than MUR. The relative population densities of both species were significantly and negatively correlated with each other. Whereas the relative population densities of MUR increased with altitude and were highest in dry habitats far from surface water, the relative population densities of RAV generally decreased with altitude and were highest in low altitude habitats close to surface water. The results of the vegetation characteristics also reflect these general trends. The divergent pattern of local and regional coexistence of these two species is discussed and can be best explained either by the existence of a spatially heterogeneous competitive environment or by independent evolutionary pathways in different historic environments. PMID- 19676111 TI - Evaluation of dalbavancin as chiral selector for HPLC and comparison with teicoplanin-based chiral stationary phases. AB - Dalbavancin is a new compound of the macrocyclic glycopeptide family. It was covalently linked to 5 microm silica particles using two different binding chemistries. Approximately 250 racemates including (a) heterocyclic compounds, (b) chiral acids, (c) chiral amines, (d) chiral alcohols, (e) chiral sulfoxides and sulfilimines, (f) amino acids and amino acid derivatives, and (g) other chiral compounds were tested on the two new chiral stationary phases (CSPs) using three different mobile phases. As dalbavancin is structurally related to teicoplanin, the same set of chiral compounds was screened on two commercially available teicoplanin CSPs for comparison. The dalbavancin CSPs were able to separate some enantiomers that were not separated by the teicoplanin CSPs and also showed improved separations for many racemates. However, there were other compounds only separated or better separated on teicoplanin CSPs. Therefore, the dalbavancin CSPs are complementary to the teicoplanin CSPs. PMID- 19676113 TI - Large-scale analysis of secondary structure changes in proteins suggests a role for disorder-to-order transitions in nucleotide binding proteins. AB - Conformational changes in proteins often involve secondary structure transitions. Such transitions can be divided into two types: disorder-to-order changes, in which a disordered segment acquires an ordered secondary structure (e.g., disorder to alpha-helix, disorder to beta-strand), and order-to-order changes, where a segment switches from one ordered secondary structure to another (e.g., alpha-helix to beta-strand, alpha-helix to turn). In this study, we explore the distribution of these transitions in the proteome. Using a comprehensive, yet highly conservative method, we compared solved three-dimensional structures of identical protein sequences, looking for differences in the secondary structures with which they were assigned. Protein chains in which such secondary structure transitions were detected, were classified into two sets according to the type of transition that is involved (disorder-to-order or order-to-order), allowing us to characterize each set by examining enrichment of gene ontology terms. The results reveal that the disorder-to-order set is significantly enriched with nucleotide binding proteins, whereas the order-to-order set is more diverse. Remarkably, further examination reveals that >22% of the purine nucleotide binding proteins include segments which undergo disorder-to-order transitions, suggesting that such transitions play an important role in this process. PMID- 19676114 TI - Crystal structure of Arabidopsis translation initiation factor eIF-5A2. PMID- 19676116 TI - Evolution of viviparity and uterine angiogenesis: vascular endothelial growth factor (VEGF) in oviparous and viviparous skinks. AB - During pregnancy, uterine vasculature of live-bearing lizards proliferates to support embryonic growth and development. Vascular endothelial growth factor (VEGF) is the most potent of a suite of growth factors responsible for uterine vascularization in mammals. We have sequenced VEGF mRNA transcripts expressed in the uterus of oviparous and viviparous Australian skinks, and compared uterine VEGF expression in nonreproductive and late-reproductive Saiphos equalis, a fossorial viviparous skink. VEGF sequences differed between phylogenetic groups of skinks, rather than oviparous and viviparous skinks. Two transcripts were identified in the uterus of each species that had the same splice sites as human VEGF(165) and VEGF(189). A third transcript, found only in uterine and testis tissue from S. equalis, had the same splice sites as human VEGF(111). This is the first natural expression of VEGF(111), previously found only in human cultured cells subjected to environmental stress. All the three VEGF transcripts identified showed higher expression in uterus from late-reproductive S. equalis than nonreproductive females. The different angiogenic properties of VEGF transcripts provide a mechanism that may produce the variety of placental complexities observed in viviparous skinks. The presence of VEGF(111) in S. equalis may be an opportunity to investigate the function of this unique transcript in a whole animal system. PMID- 19676117 TI - Fatal CMV pneumonitis in a lymphoma patient treated with rituximab. PMID- 19676115 TI - Structural insights into recognition of beta2-glycoprotein I by the lipoprotein receptors. AB - The interactions of beta2 glycoprotein I (B2GPI) with the receptors of the low density lipoprotein receptor (LDLR) family are implicated in the clearance of negatively charged phospholipids and apoptotic cells and, in the presence of autoimmune anti-B2GPI antibodies, in cell activation, which might play a role in the pathology of antiphospholipid syndrome (APS). The ligand-binding domains of the lipoprotein receptors consist of multiple homologous LA modules connected by flexible linkers. In this study, we investigated at the atomic level the features of the LA modules required for binding to B2GPI. To compare the binding interface in B2GPI/LA complex to that observed in the high-resolution co-crystal structure of the receptor associated protein (RAP) with a pair of LA modules 3 and 4 from the LDLR, we used LA4 in our studies. Using solution NMR spectroscopy, we found that LA4 interacts with B2GPI and the binding site for B2GPI on the (15)N-labeled LA4 is formed by the calcium coordinating residues of the LA module. We built a model for the complex between domain V of B2GPI (B2GPI-DV) and LA4 without introducing any experimentally derived constraints into the docking procedure. Our model, which is in the agreement with the NMR data, suggests that the binding interface of B2GPI for the lipoprotein receptors is centered at three lysine residues of B2GPI-DV, Lys 308, Lys 282, and Lys317. PMID- 19676118 TI - Predicting survival in adults with invasive aspergillosis during therapy for hematological malignancies or after hematopoietic stem cell transplantation: Single-center analysis and validation of the Seattle, French, and Strasbourg prognostic indexes. AB - In this retrospective monocenter study, we analyzed the outcomes of 130 adult hematological patients who developed a proven (n = 23), probable (n = 71), and possible (n = 36) invasive aspergillosis (IA) in a 13-year period. Forty-nine patients (38%) were recipients of an allogeneic hematopoietic stem cell transplantation (AlloHSCT). The main goal of the study was the identification of prognostic factors for 4-month aspergillosis free survival (AFS) and overall survival (OS). IA was identified as the main cause of death in 27/49 recipients of an AlloHSCT (55%) and 28/81 nontransplanted patients (35%). Diagnosis of IA at or before 2000 had a negative impact in both 4-month AFS and 4-month OS in the entire group. In multivariate analysis performed separately for nontransplanted and allo-HSCT patients, five variables (excluding the year of diagnosis) decreased 4-month AFS: (i) impairment of one organ function (OF), (ii) impairment of two or more OFs (two points), (iii) disseminated IA, (iv) neutropenia lasting more than 10 days (non-AlloHSCT group only) or monocytopenia (<0.1 x 10(9)/l) [AlloHSCT group only], and (v) high-dose steroids (non-AlloHSCT group only) or an alternative donor (AlloHSCT group only). According to the number of adverse risk factors, three prognostic subgroups were defined in non-transplanted and alloHSCT patients with good (97% and 78% AFS), intermediate (73% and 32% AFS) and poor prognosis (20% and 11% AFS) of IA [P < 0.01]. In addition, we validated the French and Seattle prognostic indexes for allo-HSCT recipients and the Strasbourg model for all hematological patients with IA. PMID- 19676119 TI - The revival of clinical wisdom: The case of oral anticoagulation management. PMID- 19676120 TI - The systems biology-based argument for taking a bold step in chemoprophylaxis of sickle vasculopathy. PMID- 19676121 TI - Balancing work and life: an interview with Connie Eaves. PMID- 19676122 TI - Balancing work and life: an interview with Kathleen Sakamoto. PMID- 19676123 TI - Doublecortin and CaM kinase-like-1 and leucine-rich-repeat-containing G-protein coupled receptor mark quiescent and cycling intestinal stem cells, respectively. AB - It is thought that small intestinal epithelia (IE) undergo continuous self renewal primarily due to their population of undifferentiated stem cells. These stem cells give rise to transit amplifying (daughter/progenitor) cells, which can differentiate into all mature cell types required for normal gut function. Identification of stem cells in IE is paramount to fully understanding this renewal process. One major obstacle in gastrointestinal stem cell biology has been the lack of definitive markers that identify small intestinal stem cells (ISCs). Here we demonstrate that the novel putative ISC marker doublecortin and CaM kinase-like-1 (DCAMKL-1) is predominantly expressed in quiescent cells in the lower two-thirds of intestinal crypt epithelium and in occasional crypt-based columnar cells (CBCs). In contrast, the novel putative stem cell marker leucine rich-repeat-containing G-protein-coupled receptor (LGR5) is observed in rapidly cycling CBCs and in occasional crypt epithelial cells. Furthermore, functionally quiescent DCAMKL-1+ crypt epithelial cells retain bromo-deoxyuridine in a modified label retention assay. Moreover, we demonstrate that DCAMKL-1 is a cell surface expressing protein; DCAMKL-1+ cells, isolated from the adult mouse small intestine by fluorescence activated cell sorting, self-renew and ultimately form spheroids in suspension culture. These spheroids formed glandular epithelial structures in the flanks of athymic nude mice, which expressed multiple markers of gut epithelial lineage. Thus, DCAMKL-1 is a marker of quiescent ISCs and can be distinguished from the cycling stem/progenitors (LGR5+). Moreover, DCAMKL-1 can be used to isolate normal small intestinal stem cells and represents a novel research tool for regenerative medicine and cancer therapy. PMID- 19676124 TI - Adipose-derived mesenchymal stem cells ameliorate chronic experimental autoimmune encephalomyelitis. AB - Mesenchymal stem cells (MSCs) represent a promising therapeutic approach for neurological autoimmune diseases; previous studies have shown that treatment with bone marrow-derived MSCs induces immune modulation and reduces disease severity in experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis. Here we show that intravenous administration of adipose-derived MSCs (ASCs) before disease onset significantly reduces the severity of EAE by immune modulation and decreases spinal cord inflammation and demyelination. ASCs preferentially home into lymphoid organs but also migrates inside the central nervous system (CNS). Most importantly, administration of ASCs in chronic established EAE significantly ameliorates the disease course and reduces both demyelination and axonal loss, and induces a Th2-type cytokine shift in T cells. Interestingly, a relevant subset of ASCs expresses activated alpha 4 integrins and adheres to inflamed brain venules in intravital microscopy experiments. Bioluminescence imaging shows that alpha 4 integrins control ASC accumulation in inflamed CNS. Importantly, we found that ASC cultures produce basic fibroblast growth factor, brain-derived growth factor, and platelet-derived growth factor AB. Moreover, ASC infiltration within demyelinated areas is accompanied by increased number of endogenous oligodendrocyte progenitors. In conclusion, we show that ASCs have clear therapeutic potential by a bimodal mechanism, by suppressing the autoimmune response in early phases of disease as well as by inducing local neuroregeneration by endogenous progenitors in animals with established disease. Overall, our data suggest that ASCs represent a valuable tool for stem cell-based therapy in chronic inflammatory diseases of the CNS. PMID- 19676125 TI - Impact of high-dose peginterferon alfa-2A on virological response rates in patients with hepatitis C genotype 1: a randomized controlled trial. AB - This study tested the hypothesis that high-dose peginterferon alfa-2a (PEG IFNalpha-2a) for the first 12 weeks would increase early and sustained virological response (SVR) rates in patients with chronic hepatitis C genotype 1. Eight hundred ninety-six patients were randomized 1:1 to 360 microg (n = 448) or 180 microg (n = 448) PEG-IFNalpha-2a weekly plus ribavirin at 1000-1200 mg/day for 12 weeks, followed by 36 weeks of 180 microg PEG-IFNalpha-2a weekly plus ribavirin at 1000-1200 mg/day with 871 patients evaluable for the intention-to treat analysis. Virological responses were assessed by TaqMan (limit of detection 15 IU/mL) at week 4, 8, 12, 24, 48 (end of therapy), and 24 weeks following therapy (SVR). Undetectable hepatitis C virus RNA rates were significantly higher among patients receiving high-dose induction therapy at week 4 (36% versus 26%, P < 0.005), week 8 (61% versus 50%, P < 0.005), and week 12 (74% versus 62%, P < 0.005). However, SVR was not significantly different between patients receiving high-dose (53%) and standard (50%) therapy. Significant baseline prognostic factors for SVR included age, sex, race, histological stage, and viral load. SVR was considerably higher among patients with no or minimal fibrosis (64% and 60%, respectively) compared to those with severe fibrosis/cirrhosis (28% and 24%, respectively). The frequency of serious adverse events and drug discontinuations were similar in both groups, whereas PEG-IFN dose modification, weight and appetite reduction, and grade IV neutropenia were significantly higher in the induction arm. CONCLUSION: Induction dosing with 360 microg/week PEG-IFNalpha-2a for 12 weeks was well tolerated and enhanced early virological response but not SVR rates. The high SVR rates in patients with minimal fibrosis highlight the benefit of early treatment in patients with hepatitis C virus genotype 1. PMID- 19676126 TI - Pan-caspase inhibitor VX-166 reduces fibrosis in an animal model of nonalcoholic steatohepatitis. AB - Nonalcoholic fatty liver disease (NAFLD) is a potentially progressive liver disease that culminates in cirrhosis. Cirrhosis occurs more often in individuals with nonalcoholic steatohepatitis (NASH) than in those with steatosis (nonalcoholic fatty liver [NAFL]). The difference between NAFL and NASH is the extent of hepatocyte apoptosis, which is more extensive in NASH. Because phagocytosis of apoptotic cells activates hepatic stellate cells (HSCs), we examined the hypothesis that a pan-caspase inhibitor, VX-166, would reduce progression of fibrosis in a mouse model of NASH. Male db/db mice were fed methionine/choline-deficient (MCD) diets to induce NASH and liver fibrosis. Mice were gavaged once daily with either the pan-caspase inhibitor VX-166 (6 mg/kg/d; Vertex, Abingdon, UK) or vehicle only and sacrificed at 4 or 8 weeks. Treatment with an MCD diet increased alanine aminotransferase (ALT), caspase-3 activity, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) positive cells, NASH, and fibrosis. Treatment of MCD-fed mice with VX-166 decreased active caspase-3, TUNEL-positive cells, and triglyceride content (P < 0.05). However, ALT levels were similar in VX-166-treated mice and vehicle treated controls. Histological findings also confirmed that both groups had comparable liver injury (NAFLD activity score >or=6). Nevertheless, VX-166 treated MCD-fed mice demonstrated decreased alpha-smooth muscle actin expression (4 weeks, P < 0.05; 8 weeks, P < 0.005) and had reduced hepatic levels of collagen 1alpha1 messenger RNA (8 weeks, P < 0.05). Hydroxyproline content and Sirius red staining of VX-166-treated livers confirmed decreases in fibrosis. CONCLUSION: Inhibiting hepatic apoptosis suppresses the development of fibrosis in mice with NASH. Beneficial effects on liver fibrosis were associated with reductions in hepatic steatosis, but occurred without obvious improvement in liver injury. These findings are consistent with evidence that apoptosis triggers HSC activation and liver fibrosis and suggest that caspase inhibitors may be useful as an antifibrotic NASH therapy. PMID- 19676127 TI - A seven-gene signature (cirrhosis risk score) predicts liver fibrosis progression in patients with initially mild chronic hepatitis C. AB - Fibrosis progression is the main determinant of liver disease outcome in chronic hepatitis C, being influenced by environmental and host factors. Recently, a cirrhosis risk score (CRS) based on seven single-nucleotide polymorphisms was proposed as genetic predictor of cirrhosis in hepatitis C. To assess the role of CRS in predicting fibrosis progression in patients with initially no or minimal to moderate fibrosis, we investigated 271 untreated patients with chronic hepatitis C having initial liver biopsy showing METAVIR stage F0 (n = 104), F1 (n = 101), or F2 (n = 59) who had been followed up without antiviral therapies for at least 60 months (mean 108.5 +/- 71.5 months) and had a liver biopsy at the end of this observation period. Of these, 24.4% showed no histologic progression, 75.6% progressed by at least one stage, 45.0% progressed by at least two stages, and 10.3% progressed by more than two stages. The mean CRS was significantly higher (P = 0.005) in patients with fibrosis progression compared with those without progression, and this difference was particularly evident (P = 0.002) with F0 on initial biopsy. Mean CRS scores were not associated with degree of fibrosis progression. The relative risk of fibrosis progression increased with increasing CRS values. This association was significant in males but not in females and was most evident in males with F0 at initial biopsy (odds ratio 16.5, 95% confidence interval 1.6-166; P= 0.02) in the presence of high CRS. Multivariate analysis confirmed the significant association of CRS score with fibrosis progression. The predictive value of CRS was confirmed in hepatitis C virus patients admitting significant alcohol intake. CONCLUSION: Host genetics defined by CRS predict fibrosis progression in males with initially mild chronic hepatitis C and may become a useful parameter for prognostic evaluation and treatment decision. PMID- 19676129 TI - HuR regulates gap junctional intercellular communication by controlling beta catenin levels and adherens junction integrity. AB - Gap junctional intercellular communication (GJIC) plays a critical role in the regulation of tissue homeostasis and carcinogenesis and is modulated by the levels, subcellular localization, and posttranslational modification of gap junction proteins, the connexins (Cx). Here, using oval cell-like rat liver epithelial cells, we demonstrate that the RNA-binding protein HuR promotes GJIC through two mechanisms. First, HuR silencing lowered the levels of Cx43 protein and Cx43 messenger RNA (mRNA), and decreased Cx43 mRNA half-life. This regulation was likely due to the direct stabilization of Cx43 mRNA by HuR, because HuR associated directly with Cx43 mRNA, a transcript that bears signature adenylate uridylate-rich (AU-rich) and uridylate-rich (U-rich) sequences in its 3' untranslated region. Second, HuR silencing reduced both half-life and the levels of beta-catenin mRNA, also a target of HuR; accordingly, HuR silencing lowered the levels of whole-cell and membrane-associated beta-catenin. Coimmunoprecipitation experiments showed a direct interaction between beta catenin and Cx43. Small interfering RNA (siRNA)-mediated depletion of beta catenin recapitulated the effects of decreasing HuR levels: it attenuated GJIC, decreased Cx43 levels, and redistributed Cx43 to the cytoplasm, suggesting that depletion of beta-catenin in HuR-silenced cells contributed to lowering Cx43 levels at the membrane. Finally, HuR was demonstrated to support GJIC after exposure to a genotoxic agent, doxorubicin, or an inducer of differentiation processes, retinoic acid, thus pointing to a crucial role of HuR in the cellular response to stress and in physiological processes modulated by GJIC. CONCLUSION: HuR promotes gap junctional intercellular communication in rat liver epithelial cells through two related regulatory processes, by enhancing the expression of Cx43 and by increasing the expression of beta-catenin, which, in turn, interacts with Cx43 and is required for proper positioning of Cx43 at the plasma membrane. PMID- 19676130 TI - Impact of pegylated interferon and ribavirin on morbidity and mortality in patients with chronic hepatitis C and normal aminotransferases in France. AB - Clinicians continue to raise questions concerning the necessity of treating chronic hepatitis C virus (HCV)-infected patients with normal alanine aminotransferase (N-ALT), in light of their slower progression to cirrhosis than patients with elevated alanine aminotraferase (E-ALT). This study was undertaken to predict the impact of pegylated interferon (IFN) and ribavirin on HCV-related morbidity and mortality in patients with N-ALT. A previous Markov model was adapted to separately simulate patients with N-ALT (30%) and those with E-ALT (70%). The model estimates fibrosis progression rates according to age, sex, and whether ALT levels are normal or elevated, assuming that patients with E-ALT have a 2.6 times higher progression than those with N-ALT. It takes into account improvement in HCV screening and treatment and competitive mortality. We assumed that N-ALT patients were treated 80% less frequently between 2002 and 2004 and 70% less frequently from 2005 on, as obtained in real life from three multicentric cohorts (Hepatys, Adequation, Persee). Antiviral treatment of HCV infected populations might reduce 2008-2025 HCV-related morbidity and mortality by 34,200 cases of cirrhosis (36%, 33,000-35,000), 22,400 complications (28%, 21,000-23,000) and 17,500 deaths (25%, 17,000-18,000), including 3000 cases of cirrhosis (22%, 2000-5000), 1200 complications (15%, 1000-1700), and 1000 deaths (14%, 900-1300) in the N-ALT population, despite a probability of receiving treatment that is three to five times less in this population. If N-ALT patients are treated at the same proportions as those with E-ALT, morbidity and mortality could be further reduced by 1400 cases of cirrhosis (13%, 1200-2200), 600 complications (9%, 600-1000), and 500 deaths (9%, 500-800). CONCLUSION: Treatment of N-ALT patients would decrease HCV morbidity and mortality. These patients should be considered candidates for treatment just as others are. PMID- 19676128 TI - Coffee intake is associated with lower rates of liver disease progression in chronic hepatitis C. AB - Higher coffee consumption has been associated inversely with the incidence of chronic liver disease in population studies. We examined the relationship of coffee consumption with liver disease progression in individuals with advanced hepatitis C-related liver disease. Baseline coffee and tea intake were assessed in 766 participants of the Hepatitis C Antiviral Long-Term Treatment against Cirrhosis (HALT-C) trial who had hepatitis C-related bridging fibrosis or cirrhosis on liver biopsy and failed to achieve a sustained virological response to peginterferon plus ribavirin treatment. Participants were followed for 3.8 years for clinical outcomes and, for those without cirrhosis, a 2-point increase in Ishak fibrosis score on protocol biopsies. At baseline, higher coffee consumption was associated with less severe steatosis on biopsy, lower serum aspartate aminotransferase (AST)/alanine aminotransferase (ALT) ratio, alpha fetoprotein, insulin, and homeostatic model assessment (HOMA2) score, and higher albumin (P < 0.05 for all). Two hundred thirty patients had outcomes. Outcome rates declined with increasing coffee intake: 11.1/100 person-years for none, 12.1 for less than 1 cup/day, 8.2 for 1 to fewer than 3 cups/day, and 6.3 for 3 or more cups/day (P-trend = 0.0011). Relative risks (95% confidence intervals) were 1.11 (0.76-1.61) for less than 1 cup/day; 0.70 (0.48-1.02) for 1 to fewer than 3 cups/day; and 0.47 (0.27-0.85) for 3 or more cups/day (P-trend = 0.0003) versus not drinking. Risk estimates did not vary by treatment assignment or cirrhosis status at baseline. Tea intake was not associated with outcomes. CONCLUSION: In a large prospective study of participants with advanced hepatitis C-related liver disease, regular coffee consumption was associated with lower rates of disease progression. PMID- 19676132 TI - Does hepatitis B surface antigen kinetics, including both decline and reaching a threshold, improve prediction of response? PMID- 19676131 TI - Targeting cadherin-17 inactivates Wnt signaling and inhibits tumor growth in liver carcinoma. AB - Hepatocellular carcinoma (HCC) is a lethal malignancy for which there are no effective therapies. To develop rational therapeutic approaches for treating this disease, we are performing proof-of-principle studies targeting molecules crucial for the development of HCC. Here, we show that cadherin-17 (CDH17) adhesion molecule is up-regulated in human liver cancers and can transform premalignant liver progenitor cells to produce liver carcinomas in mice. RNA interference mediated knockdown of CDH17 inhibited proliferation of both primary and highly metastatic HCC cell lines in vitro and in vivo. The antitumor mechanisms underlying CDH17 inhibition involve inactivation of Wnt signaling, because growth inhibition and cell death were accompanied by relocalization of beta-catenin to the cytoplasm and a concomitant reduction in cyclin D1 and an increase in retinoblastoma. CONCLUSION: Our results identify CDH17 as a novel oncogene in HCC and suggest that CDH17 is a biomarker and attractive therapeutic target for this aggressive malignancy. PMID- 19676133 TI - A novel nonsynonymous variant of matrix metalloproteinase-7 confers risk of liver cirrhosis. AB - Liver cirrhosis is characterized by progressive accumulation of extracellular matrix following chronic liver injuries. In the extracellular space, the constant turnover of liver matrix is regulated by the matrix metalloproteinase (MMP) class of enzyme. To assess whether genetic variations in MMP would result in diversity of liver cirrhosis, a case-control study of 320 patients with hepatocellular carcinoma, with or without cirrhosis, was conducted. Ten single-nucleotide polymorphism markers from four potential fibrosis-associated genes were selected for genotyping. Among these genes, a nonsynonymous single-nucleotide polymorphism which generates the variation of Gly-137 and Asp-137 in the MMP-7 gene was found to be strongly associated with the development of liver cirrhosis. In contrast to MMP-7(Gly-137) that predominantly secretes out into the cell culture medium, the cirrhosis-associated MMP-7(Asp-137) variant is preferentially localized on the extracellular membranes where it exerts its proteolytic activity on pericellular substrates. Functional analysis demonstrated an increased ability of the MMP 7(Asp-137) variant to associate with the cell surface CD151 molecule. In wound healing and Boyden chamber assays, cell motility was specifically enhanced with the expression of MMP-7(Asp-137) as compared to the cells expressing MMP-7(Gly 137). These results demonstrate that the MMP-7(Asp-137) variant confers a gain-of function phenotype for MMP-7. CONCLUSION: We have identified a novel genetic association of MMP-7(Asp-137) variant with liver cirrhosis in patients with hepatocellular carcinoma. Whether the MMP-7 variant can be a new marker for liver cirrhosis will be further studied. PMID- 19676136 TI - Triple-negative breast cancer rate is triply high in black women. PMID- 19676135 TI - Advanced periductal fibrosis from infection with the carcinogenic human liver fluke Opisthorchis viverrini correlates with elevated levels of interleukin-6. AB - More than 750 million people are at risk of infection with foodborne liver flukes. Opisthorchis viverrini is considered among the most important of these parasites, due to its strong association with cholangiocarcinoma (CCA). O. viverrini infection results in a chronic inflammatory challenge to the host, which can lead to advanced, pathogen-specific disease sequelae including obstructive jaundice, hepatomegaly, cholecystitis, as well as CCA. However, before disease sequelae are apparent, important inflammatory changes to the liver can be detected early during O. viverrini infection. In a case-control study involving 328 men and women with O. viverrini infection, we determined the presence of advanced periductal fibrosis in asymptomatic, O. viverrini-infected individuals and then measured cytokine responses to O. viverrini excretory/secretory products (ES). In the 200 participants with advanced periductal fibrosis (cases), levels of interleukin-6 (IL-6) to O. viverrini ES were 8 times higher than levels of the 128 O. viverrini-infected individuals without advanced periductal fibrosis (controls). Moreover, elevated IL-6 to parasite ES was associated with increased risk of advanced periductal fibrosis by 63% in a model adjusted for sex and age. The risk of advanced periductal fibrosis was also found to increase with higher levels of IL-6: individuals in the third quartile of IL-6-ES production had a 127% higher risk of developing advanced periductal fibrosis than individuals in the first quartile of IL-6 production. O. viverrini-infected individuals with advanced periductal fibrosis showed other hepatobiliary abnormalities, including reduced gallbladder contractility and the presence of gallbladder sludge. CONCLUSION: These data strongly implicate a role for parasite-specific IL-6 in the pathogenesis of advanced periductal fibrosis in opisthorchiasis, with possible links to other hepatobiliary abnormalities, including CCA. PMID- 19676134 TI - Deletion of interleukin-12p40 suppresses autoimmune cholangitis in dominant negative transforming growth factor beta receptor type II mice. AB - Our laboratory has reported that mice that express a dominant negative form of transforming growth factor beta receptor restricted to T cells (dnTGFbetaRII) develop an inflammatory biliary ductular disease with elevated serum levels of interleukin (IL)-12p40 and other proinflammatory cytokines and antimitochondrial autoantibodies (AMAs) closely resembling human primary biliary cirrhosis (PBC). We have used this mouse model to address the potential mechanisms of immunomodulation of liver disease by creating two unique genetic strains: IL 12p40 knockout (KO)-dnTGFbetaRII mice and IFN-gamma KO-dnTGFbetaRII mice. The two colonies of genetically modified mice-and, for purposes of controls, the dnTGFbetaRII mice-were monitored for liver immunopathology, AMAs, and intrahepatic cytokine production. Disease expression in the IFN-gamma KO dnTGFbetaRII mice, including liver immunopathology, were similar to those of dnTGFbetaRII mice, whereas the IL-12p40 KO-dnTGFbetaRII mice had a dramatic reduction in histological autoimmune cholangitis and significant decreases in levels of intrahepatic proinflammatory cytokines, but similar levels of AMAs compared with dnTGFbetaRII controls. CONCLUSION: These data indicate that in this mouse model of PBC, signaling by way of IL-12p40 is an essential requirement for the development of autoimmune cholangitis. The results of these studies will play an important role in identifying pathways and reagents that will selectively inhibit IL-12 signaling for the outlining of future therapeutic strategies for human PBC. PMID- 19676137 TI - Dipeptidyl peptidase (DP) 6 and DP10: novel brain proteins implicated in human health and disease. AB - Dipeptidyl peptidase (DP) 6 and DP10 are non-enzyme members of the dipeptidyl peptidase IV family, which includes fibroblast activation protein, DP8, and DP9. DP6 and DP10 proteins have been shown to be critical components of voltage-gated potassium (Kv) channels important in determining cellular excitability. The aim of this paper was to review the research to date on DP6 and DP10 structure, expression, and functions. To date, the protein region responsible for modulating Kv4 channels has not been conclusively identified and the significance of the splice variants has not been resolved. Resolution of these issues will improve our overall knowledge of DP6 and DP10 and lead to a better understanding of their role in diseases, such as asthma and Alzheimer's disease. PMID- 19676138 TI - Role of dipeptidyl peptidase IV (DP IV)-like enzymes in T lymphocyte activation: investigations in DP IV/CD26-knockout mice. AB - BACKGROUND: Dipeptidyl peptidase IV (DP IV, CD26) and DP IV-like enzymes, such as dipeptidyl peptidase II (DP II), dipeptidyl peptidase 8 (DP8), and dipeptidyl peptidase 9 (DP9), have been recognized to regulate T lymphocyte activation. Lys[Z(NO2)]-thiazolidide (LZNT) and Lys[Z(NO2)]-pyrrolidide (LZNP), non-selective inhibitors of DP IV-like activity known to target DP IV as well as DP II, DP8, and DP9, suppress T lymphocyte proliferation in vitro. Moreover, these inhibitors are capable of attenuating the severity of autoimmune diseases, such as experimental autoimmune encephalomyelitis, the animal model of multiple sclerosis, and experimental arthritis, a model of human rheumatoid arthritis, in vivo, particularly in combination with inhibitors of aminopeptidase N (APN, CD13) enzymatic activity. METHODS: Here, we studied the influence of non-selective and selective inhibitors of DP IV-like enzymes on DNA synthesis in mitogen-stimulated splenocytes from wild-type C57BL/6 mice and DP IV/CD26-knockout (DP IV/CD26-KO) mice. RESULTS: LZNT and LZNP, the non-selective inhibitors of DP IV-like activity, suppressed the DNA synthesis in stimulated splenocytes from wild-type and DP IV/ CD26-KO mice to a comparable extent. Further, a selective inhibitor of DP8/DP9 activity was capable of suppressing DNA synthesis in mitogen-stimulated splenocytes of both wild-type and knockout mice to the same extent. In contrast, selective inhibitors of DP IV and DP II lacked this suppressive activity. CONCLUSIONS: Our data support the hypothesis that DP8 and/or DP9 represent additional pharmacological targets for the suppression of T cell proliferation and for anti-inflammatory therapy. PMID- 19676139 TI - A selected pre-amplification strategy for genetic analysis using limited DNA targets. AB - BACKGROUND: Limited DNA resources or limited DNA targets in predominant backgrounds for genetic tests can lead to misdiagnosis. We developed a strategy to selectively increase the amount of minor targets through a specific pre amplification procedure. METHODS: We used the model of circulating cell free (ccf) male fetal DNA as a minor target in the predominant maternal plasma DNA to evaluate the strategy. The sex determining region (SRY) locus on the Y chromosome was used to identify ccf fetal DNA, and the human glyceraldehydes-3-phosphate dehydrogenase (GAPDH) gene was used to identify ccf total DNA in maternal plasma. We selectively pre-amplified the minor target SRY locus using the Expand Long Template PCR system and assessed the efficiency of the pre-amplification by real time PCR, for both SRY and GAPDH, to compare the quantities of pre-amplified fetal DNA with those of maternal total DNA without pre-amplification. RESULTS: The selected pre-amplification increased the amount of ccf fetal DNA dramatically (Wilcoxon test: p =0.000, the fold change = 11,596). After selected preamplification, a proportion of 2.19% of the ccf fetal minor part in the predominant maternal component was changed up to 25,334%. The increased amounts of ccf fetal DNA found with the pre-amplification are not correlated to the amounts found without the procedure (r=-0.017, p=0.949). CONCLUSIONS: This strategy may be useful in genetic analysis with limited DNA resources and limited DNA targets in predominant background molecules. However, this approach is not suitable for quantitative assessments, due to the fact that quantitative imbalanced amplification was observed as a result of the pre-amplification procedure. PMID- 19676140 TI - Automated cell disruption is a reliable and effective method of isolating RNA from fresh snap-frozen normal and malignant oral mucosa samples. AB - BACKGROUND: This study compared automated vs. manual tissue grinding in terms of RNA yield obtained from oral mucosa biopsies. METHODS: A total of 20 patients undergoing uvulectomy for sleep-related disorders and 10 patients undergoing biopsy for head and neck squamous cell carcinoma were enrolled in the study. Samples were collected, snap-frozen in liquid nitrogen, and divided into two parts of similar weight. Sample grinding was performed on one sample from each pair, either manually or using an automated cell disruptor. The performance and efficacy of each homogenization approach was compared in terms of total RNA yield (spectrophotometry, fluorometry), mRNA quantity [densitometry of specific TP53 amplicons and TP53 quantitative reverse-transcribed real-time PCR (qRT-PCR)], and mRNA quality (functional analysis of separated alleles in yeast). RESULTS: Although spectrophotometry and fluorometry results were comparable for both homogenization methods, TP53 expression values obtained by amplicon densitometry and qRT-PCR were significantly and consistently better after automated homogenization (p<0.005) for both uvula and tumor samples. Functional analysis of separated alleles in yeast results was better with the automated technique for tumor samples. CONCLUSIONS: Automated tissue homogenization appears to be a versatile, quick, and reliable method of cell disruption and is especially useful in the case of small malignant samples, which show unreliable results when processed by manual homogenization. PMID- 19676141 TI - Serum free light chain immunoassay as an adjunct to serum protein electrophoresis and immunofixation electrophoresis in the detection of multiple myeloma and other B-cell malignancies. AB - BACKGROUND: Protein and immunofixation electrophoresis of serum and urine are established as diagnostic aids for identifying monoclonal gammopathies. However, many patient sera sent to laboratories are not accompanied by urine samples and recent reports suggest the use of serum free light chain (sFLC) analysis in combination with serum protein electrophoresis (SPE) and immunofixation electrophoresis (lFE) could eliminate the need for urinalysis. The aim of the study was to assess the utility of sFLC measurement in addition to serum protein electrophoresis in the identification of patients with B-cell malignancies. METHODS: A total of 952 serum samples were analysed by serum protein electrophoresis and those with abnormal bands were analysed by immunofixation. sFLCs were measured in a retrospective manner by automated assay. RESULTS: In our study of 952 patient sera, it was found that FLC analysis identified 23 additional cases of B-cell malignancies which were missed by SPE. CONCLUSIONS: The additional malignancies identified by sFLC analysis add support for its inclusion in the routine screening protocol for B-cell malignancies. PMID- 19676142 TI - Gas chromatography/mass spectrometry screening of serum metabolomic biomarkers in hepatitis B virus infected cirrhosis patients. AB - BACKGROUND: Metabolomic studies have been applied to disease biomarkers selection. With the metabolomic technique, gas chromatography/mass spectrometry (GC/MS), human serum metabolites can be detected and identified. The purpose of this study was to investigate the serum metabolic profile of hepatitis B virus (HBV) infected cirrhosis patients and to detect disease biomarkers. METHODS: HBV infected non-cirrhosis male subjects (n=20) and HBV infected cirrhosis male patients (n=20) participated in this experiment. Serum metabolome was detected through chemical derivatization followed by GC/MS. The high-flux metabolomic data were analyzed by stepwise discriminant analysis. RESULTS: Out of the 41 metabolites detected in serum, we selected metabolites, including acetic acid, sorbitol, D-lactic acid, hexanoic acid, 1-naphthalenamine, butanoic acid, phosphoric acid, D-glucitol, and glucose, which in combination with each other could segregate the two groups. The error count was 0% for the non-cirrhosis group and 25% for the cirrhosis group. CONCLUSIONS: This technique can be used to select biomarkers for hepatic cirrhosis. PMID- 19676143 TI - Evaluation of biomarkers of exposure and potential harm in smokers, former smokers and never-smokers. AB - BACKGROUND: The objective of this study was to obtain baseline data on biomarkers of exposure (BoE) and biomarkers of potential harm (BoPH) in smokers, former smokers and never-smokers. METHODS: This was a cross-sectional study of 80 healthy male and female volunteers over 21 years old, self-selected for smoking status. Subjects were prescreened by medical staff at an independent clinical research unit, within 1 week prior to a single overnight residential visit and sample collection. RESULTS: All BoE were able to differentiate between the two smoking groups and smokers from all nonsmokers. There was a strong correlation between cigarettes smoked per day and total urinary nicotine equivalents (TNE; r=0.85). TNE correlated better with 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol levels than cigarettes smoked per day (r=0.75 and r=0.56, respectively). Of the BoPH included in this study, seven (11-dehydro-thromboxane B2, 2, 3-dinorthrom boxane B2, 8-epi prostaglandin F(2alpha), 8-hydroxy-2 deoxyguanosine, cis thymidine glycol, low-density lipoprotein cholesterol and IgG) were significantly different between the group who smoked more cigarettes per day and never-smokers. These differences became more apparent and extended to the group who smoked 10 or less cigarettes per day, when total urinary recovery values were corrected for creatinine clearance. CONCLUSIONS: While BoE clearly differentiate between groups based on self-declared smoking status, most BoPH examined could not do so in a consistent manner. The dynamics of BoPH levels are not well understood. Future studies of BoPH should eliminate potential confounding factors and increase the number of subjects to allow the investigation of genetic polymorphism in metabolic pathways. PMID- 19676144 TI - Sensitive and specific ELISA coated by TpN15-TpN17-TpN47 fusion protein for detection of antibodies to Treponema pallidum. AB - BACKGROUND: We constructed an artificial fusion gene tpN15-17-47 and then used the prokaryotic expression fusion protein rTpN15-17-47 as the coated antigen to establish a new enzyme-linked immunosorbent assay (ELISA) for serological diagnosis of syphilis. METHODS: tpN15, tpN17, and tpN47 genes were amplified separately by polymerase chain reaction (PCR) and then assembled into a fusion gene coding a trigeminy protein antigen by primer linking PCR. The target recombinant protein antigens rTpN15, rTpN17, rTpN-47, and rTpN15-17-47 were expressed and then purified antigens were immobilized on the surface of microplate wells for detecting Treponema pallidum-specific antibodies by ELISAs. RESULTS: The relative positive rate of rTpN15-17-47-ELISA in 965 serum specimens of syphilis patients was 99.5%, which was higher than that of the T. pallidum hemagglutination assay (TPHA) (98.3%) (p<0.05) and much higher than that of the rTpN15-ELISA (83.1%), the rTpN17-ELISA (84.4%), the rTpN47-ELISA (82.1%), and the toluidine red unheated serum test (TRUST) (72.2%) (p<0.01). All the ELISAs and the TPHA in detecting serum specimens from 62 cases with systemic lupus erythematosus (SLE), 86 cases with rheumatic arthritis (RA), and 250 healthy cases were negative, but the TRUST was positive in five cases with SLE, seven cases with RA, and two healthy cases. CONCLUSIONS: The rTpN15-17-47-ELISA is a sensitive and specific serological screening or a diagnostic method for syphilis in the clinical setting. PMID- 19676145 TI - Expression of heme oxygenase-1, hypoxia inducible factor-1alpha, and ubiquitin in peripheral inflammatory cells from patients with coronary heart disease. AB - BACKGROUND: The increased expression of heme oxygenase-1 content, a stress response protein, directly correlates with the incidence of coronary heart disease. Down-regulation of hypoxia inducible factor-1alpha activity, a major downstream effector of the signaling pathways activated by hypoxia, increases cell survival after hypoxia. The ubiquitin system, a non-lysosomal pathway of protein degradation, is involved in processes of coronary heart disease. The aim of this study was to investigate the expression of heme oxygenase-1, hypoxia inducible factor-1alpha, and ubiquitin in both monocytes and lymphocytes isolated from patients at the mRNA and protein levels in different stages of coronary heart disease and their possible correlation. METHODS: A total of 90 patients with coronary heart disease (30 acute myocardial infarction, 30 unstable angina pectoris, and 30 stable angina pectoris) were selected, and 30 cases with normal coronary artery served as controls. The mRNA and protein expression of heme oxygenase-1, hypoxia inducible factor-1alpha, and ubiquitin in monocytes and lymphocytes were examined by semi-quantitative reverse transcriptase polymerase chain reaction and Western blotting, respectively. RESULTS: The mRNA expression of heme oxygenase-1 and ubiquitin was associated with the severity of coronary heart disease (p<0.05). There was no significant difference in hypoxia inducible factor-1alpha mRNA expression between the coronary heart disease patients and controls. The protein expression of heme oxygenase-1, hypoxia inducible factor 1alpha, and ubiquitin was significantly stronger in patients with coronary heart disease than in controls, and the expression levels increased with the severity of the disease. There was a positive association between heme oxygenase-1 and hypoxia inducible factor-1alpha and ubiquitin, antioxidative therapy with adrenergic receptor blocker, angiotensin-converting enzyme inhibitor or statins up-regulated the expression of heme oxygenase-1 and hypoxia inducible factor 1alpha. CONCLUSIONS: These data suggest that heme oxygenase-1, hypoxia inducible factor-1alpha, and ubiquitin are involved in the development and progression of coronary heart disease and thus may be useful biomarkers for coronary heart disease. PMID- 19676146 TI - Vitamin B12 deficiency is associated with coronary artery disease in an Indian population. AB - BACKGROUND: The incidence of coronary artery disease (CAD) is increasing at an alarming rate, especially in developing countries, such as India. It is often advocated that a vegetarian lifestyle could reduce the burden of CAD. However, in spite of a majority of Indians being vegetarians, the incidence of CAD is highest in this population. This may be due to deficiency of vitamin B12, a micronutrient, sourced only from animal products. METHODS: Herein, we assessed the effect of vitamin B12 with respect to CAD in 816 individuals (368 CAD patients and 448 controls) recruited from a tertiary care center in New Delhi, India. RESULTS: We found that vitamin B12 levels were significantly lower in CAD patients than in controls (p<0.0001). Also, vegetarians were found to have significantly lower vitamin B12 concentrations (p=0.0001) and higher incidence of CAD (p=0.01). Interestingly, elevated homocysteine levels, a hallmark of vitamin B12 deficiency, was not associated with CAD. In contrast, cysteine levels were significantly higher in CAD patients than in controls (p=0.004). CONCLUSIONS: We believe that, when vitamin B12 is deficient, homocysteine is rapidly metabolized via the transsulfuration pathway leading to increased cysteine levels. PMID- 19676147 TI - Plasma osteoprotegerin is an independent risk factor for mortality and an early biomarker of coronary vascular calcification in chronic kidney disease. AB - BACKGROUND: Cardiovascular disease is the major cause of morbidity and mortality in chronic kidney disease (CKD) and early biomarkers are required which can predict disease and death in such patients. The aim of our study was to investigate if osteoprotegerin (OPG) could be a predictor of coronary artery calcification (CAC) and mortality in CKD. METHODS: A total of 77 outpatients (32 with pre-dialysis CKD and 45 undergoing hemodialysis) were followed-up during 2 years. Measurements of CAC were performed using Siemens Multidetector CT software and calcium scores were measured according to the Agatston method. RESULTS: OPG was an independent predictor of the Agatston score for CAC and correlated with the degree of CAC in pre-dialysis patients. A two-sample t-test characterized survivors as having a better glomerular filtration rate, lower Agatston scores, and lower serum levels of OPG. Kaplan-Meier survival curves separated survivors from non-survivors at plasma OPG cut-off levels of <3.1 ng/mL. A multivariable logistic regression analysis showed that OPG was an independent predictor of mortality from all causes in CKD patients. CONCLUSIONS: OPG predicted mortality in CKD patients and could be a valuable biomarker in early detection of CAC in these patients. PMID- 19676148 TI - Clinical usefulness of measuring red blood cell distribution width on admission in patients with acute coronary syndromes. AB - BACKGROUND: Conventional cardiac markers used for the biochemical diagnosis of acute coronary syndromes (ACS) have a high specificity but low sensitivity within 2-4 h of symptoms onset. The red blood cell distribution width (RDW), reflecting the size variability of circulating red blood cells, has been shown to be independently associated with an increased risk of major cardiovascular events. We assessed whether there is an association between RDW at admission and cardiac troponin T (cTnT) elevation in patients with chest pain. METHODS: We analyzed RDW values in 2304 adult patients, who were consecutively admitted over a 1-year period to the local emergency department for chest pain suggestive of ACS. In all patients, a baseline blood sample was collected for routine haematological testing, whereas cTnT was measured at baseline and after 4, 6, and 12 h. RESULTS: A total of 456 patients (19.8% of total) had ACS. These patients, all having cTnT> or =0.03 microg/L up to 12 h from admission other than ischaemic electrocardiographic changes, had higher RDW than non-ACS patients [median 15.1%, (5th-95th percentiles) 13.2%-19.0% vs. 13.5%, 12.9%-17.1%, p<0.001]. On admission, the sensitivity and specificity of cTnT were 94% (25 false negative results) and 100%, respectively. The diagnostic accuracy of RDW, as calculated by the receiver operating characteristic curve analysis, was 0.705 (p<0.001). At the cut-off value of 14%, the clinical sensitivity and specificity of RDW on admission were 79% and 50%, respectively. In 21 out of 25 patients classified as false negative for cTnT on admission, the RDW was >14%. Accordingly, the diagnostic sensitivity of the two combined measurements on admission was 99%. CONCLUSIONS: As RDW is widely available to clinicians as a part of the complete blood count, and therefore incurs no additional costs, it might be considered with other conventional cardiac markers for the risk stratification of ACS patients admitted to emergency departments. PMID- 19676150 TI - Precision performance at low levels and 99th percentile concentration of the Access AccuTnl assay on two different platforms. AB - BACKGROUND: Cardiac troponins currently represent the preferred biomarkers for the detection of myocardial necrosis. The objective of the present study was to compare the performance of the Access AccuTnl assay (Beckman Coulter) measured on two different platforms, the UniCel Dxl 800 and the Access 2 (Beckman Coulter). In particular, the serum cardiac troponin I (cTnl) concentration corresponding to 10% coefficient of variation (CV), the cTnl assay minimum detectable concentration (MDC), and the serum cTnl 99th percentile in healthy subjects were calculated. METHODS: The Access AccuTnl is a paramagnetic particle chemiluminescent immunoassay. Imprecision profiles were determined according to the Clinical and Laboratory Standards Institute EP5-A protocol using serum pools. The MDC was calculated as mean +3 SD of 20 determinations of the zero calibrator during one run. The 99th percentile was determined analyzing serum samples from 679 healthy blood donors (523 males, 156 females; 18-71 years old). RESULTS: cTnl concentrations are given in microg/L. 10% CV values (95% confidence interval, CI) were 0.0577 (0.0467-0.0750) (UniCel Dxl 800) and 0.0486 (0.0255-0.0596) (Access 2). MDC values were 0.011 (UniCel Dxl 800) and 0.012 (Access 2). The 99th percentile (95% CI) value was 0.0340 (0.0298-0.0410). CONCLUSIONS: Our data confirm the reliability of the evaluated cTnl assay and demonstrate the comparability of the cTnl values between the platforms studied. PMID- 19676149 TI - Evaluation of the BD Vacutainer PST II blood collection tube for special chemistry analytes. AB - BACKGROUND: The performance of the BD Vacutainer PST II Tube for testing of special chemistry analytes was evaluated in comparison to serum and heparin plasma non-gel blood collection tubes. The comparison tubes included the BD Vacutainer Serum Plus Tube, the BD Vacutainer Lithium Heparin Plus Tube, and the BD Vacutainer Lithium Heparin Glass Tube. METHODS: Tubes were drawn by routine venipuncture from 42 subjects according to a randomized draw order. Tubes were processed and centrifuged according to recommended handling procedures. Serum and plasma from the comparison tubes were aliquoted to secondary containers prior to analysis. Specimens were then tested for selected special chemistry analytes at two time intervals (initial time and after 24 h storage). Analytes tested included thyroid stimulating hormone, free thyroxine, total thyroxine, follicle stimulating hormone, luteinizing hormone, ferritin, cortisol, vitamin B12, folate, and testosterone. The data were collected and analyzed by analysis of variance and mean bias comparisons. RESULTS: The performance of the BD Vacutainer PST II Tube was considered to be clinically equivalent to all three comparison tubes for all assays. Test results from the BD Vacutainer PST II Tube and from aliquots from the three comparison tubes at 24 h were considered to be clinically equivalent to those at initial time for all assays. CONCLUSIONS: The BD Vacutainer PST II Tube provided clinically equivalent results to serum and plasma non-gel tubes and good storage stability for the assays evaluated without the need to aliquot. PMID- 19676151 TI - The European Register of Specialists in Clinical Chemistry and Laboratory Medicine: Code of Conduct, Version 2--2008. AB - In 1997, the European Communities Confederation of Clinical Chemistry and Laboratory Medicine (EC4) set up a Register for European Specialists in Clinical Chemistry and Laboratory Medicine. The operation of the Register is undertaken by a Register Commission (EC4RC). During the last 10 years, more than 2000 specialists in Clinical Chemistry and Laboratory Medicine have joined the Register. In 2007, EC4 merged with the Federation of European Societies of Clinical Chemistry and Laboratory Medicine (FESCC) to form the European Federation of Clinical Chemistry and Laboratory Medicine (EFCC). A Code of Conduct was adopted in 2003 and a revised and updated version, taking account particularly of the guidelines of the Conseil Europeen des Professions Liberales (CEPLIS) of which EFCC is a member, is presented in this article. The revised version was approved by the EC4 Register Commission and by the EFCC Executive Board in Paris on 6 November, 2008. PMID- 19676152 TI - FOXO1 regulates the expression of 4E-BP1 and inhibits mTOR signaling in mammalian skeletal muscle. PMID- 19678402 TI - Help for housewives: domestic service and the reconstruction of domesticity in Britain, 1940-50. AB - This article argues that examining the debates over domestic science that took place in the late 1940s can produce insights into the ways in which gender, class and domesticity were inextricably linked and understood. The article focuses on three specific examples: a social survey of domestic service produced privately, a government report on the domestic organization of private households, and a radio debate broadcast by the BBC Home Service. PMID- 19678403 TI - ["The axis of wickedness--or necessary collaborator?"]. PMID- 19678404 TI - [Myths block good patient information]. PMID- 19678405 TI - [Comments to hospital standardised mortality ratios]. PMID- 19678406 TI - [Physicians and climate changes]. PMID- 19678407 TI - [Depriciation of physicians handbooks]. PMID- 19678409 TI - [Current practice of tuberculosis diagnosis, its treatment and challenge in Japan]. PMID- 19678408 TI - [Exercise, physical activities and public health (15) To produce desirable effects of exercise and risk management]. PMID- 19678410 TI - [Determination of psychosocial factors (2) Psychological characteristics II. Validity]. PMID- 19678411 TI - Re: Some positives in animal welfare. PMID- 19678412 TI - Mutagenic activity of potentiated antibodies to erythropoietin. AB - Single and repeated administration of ultralow doses of antibodies to erythropoietin did not increase the count of aberrant metaphases in bone marrow cells of BALB/c mice and were not genotoxic for Drosophila melanogaster wing cells in the test of somatic mosaicism. PMID- 19678413 TI - Speak as one united voice and children will listen! PMID- 19678414 TI - Narcolepsy. Interview by Norman Sussman. PMID- 19678415 TI - Alternate day dosing of rosuvastatin: potential usefulness in statin-intolerant patients. PMID- 19678416 TI - Women's work in offices and the preservation of men's "breadwinning" jobs in early twentieth-century Glasgow. AB - As Britain's industrial economy matured and the volume of administrative work increased, different kinds of clerical jobs and clerical careers became possible. Using examples from a variety of small- to medium-sized enterprises in Glasgow, this article will describe how the main functions of administrative work - financial, secretarial and managerial - were divided both horizontally and vertically in order to preserve secure, well-paid, "breadwinning" jobs for men, leaving routine secretarial work for women. The isolation of women in all-women enclaves carrying out shorthand and typing work and the subsequent devaluation of these as kinds of work were of primary importance in the creation of office work that was explicitly women's work. PMID- 19678417 TI - Life into art and art into life: visualising the aesthetic woman or "high art maiden" of the Victorian "Renaissance.". AB - There has been a great deal of research concerning the effect of aestheticism on men in the late nineteenth century, especially in terms of their sexuality, but its impact on women has been largely dismissed as unnecessary, as women were naturally "feminine". This article examines whether "the self-absorption and heightened emotional life of Aestheticism" (J.B. Bullen, 1998 [Oxford: Oxford University Press]) was tolerable in women or if it was leading to disease, decay and corruption. Aestheticism's insistence on individuality and self-discovery were to have important consequences for women, fuelling the desire for a life of their own rather than solely service to others. PMID- 19678418 TI - Proceedings of the 2008 Perfusion Downunder Winter Meeting, August 7-10, 2008, Queenstown, New Zealand. PMID- 19678419 TI - Lydia Becker's "School for Science": a challenge to domesticity. AB - Lydia Becker (1827-1890) is known as a leader of the Women's Suffrage Movement but little is known about her work to include women and girls in science. Before her energy was channelled into politics, she aimed to have a scientific career. Mid-Victorian Britain was a period in which women's intellect and potential were widely debated, and in which the dominant ideology was that their primary role in life was that of wife and mother. Science was widely regarded as a "masculine" subject which women were deliberately discouraged from studying. The author concentrates on the two main areas in which important contributions were made, the British Association for the Advancement of Science, and the Manchester School Board. PMID- 19678422 TI - Abstracts of the XVIII International HIV Drug Resistance Workshop: Basic Principles and Clinical Implications, June 9-13, 2009, Fort Myers, Florida, USA. PMID- 19678421 TI - Experiences of women estate workers during the apprenticeship period in St. Vincent, 1834-38: the transition from slavery to freedom. AB - In 1834, the British Government abolished slavery in the British Caribbean. In order to appease slave owners, the Government awarded them 20 million compensation money and initiated an apprenticeship period. The aim of the apprenticeship period was to provide a transition from slave to wage labour and tie the ex-slaves to their owners for a further six years. The apprenticeship period was marred by repressive acts by planters against their workers. Women were especially singled out for abuse and lost many of the rights they had gained during slavery. Apprenticeship was eventually abandoned in 1838. Stories of excessive cruelty convinced the colonial authorities that the working relationship between apprentices and managers within the sugar estates had not improved. This article examines the experiences of women apprentices in St Vincent to highlight the indignities that they faced. It also explores the actions that some women employed to improve their working conditions. Previous studies of the Caribbean have mainly focused on the slavery period or the twentieth century. There has so far been little attention paid to the lives of African-Caribbean women immediately after the abolition of slavery. This study helps to explain why so many women withdrew from estate work as soon as they were fully free and chose instead to concentrate on growing and marketing provision crops. During this transitionary period, it became clear to women that working conditions on the estates would not improve for them. The plantation managers' inability to adjust to free labour resulted in a significant withdrawal of female labourers after 1 August 1838. PMID- 19678423 TI - "Private rooms and back doors in abundance": the illusion of privacy in pornography in seventeenth-century England. AB - This article discusses pornography in seventeenth-century England in relation to the public/private debate. The seventeenth century is seen as bordering a shift from a communal, "public" style of living to a private, confined, inward-looking sensibility discernible from the eighteenth century. The increasing availability and development of a market for pornography, which goes hand in hand with the expansion of print culture, is seen as part of this shift, as it seems to exemplify par excellence the private consumption of printed material for private pleasure. The pornographic literature of the seventeenth century repeatedly invokes the idea of a public/private distinction to produce an eroticised narrative in which the illusion of privacy is constantly breached by the pursuit of voyeuristic pleasure. PMID- 19678424 TI - [Organic agriculture in Austria - the end of an illusion? On the occasion of the 80th birthday of Ulrich Planck]. PMID- 19678425 TI - [The concept and the ideal of sustainable land development in Germany: documentation and an interim assessment]. PMID- 19678426 TI - [Treatment advanced directives: a moral reflection to exceed the principles. The significance of virtue ethics]. PMID- 19678427 TI - [The biological will: notes to expand the democracy spaces in contemporary society]. PMID- 19678428 TI - [Provisioning urban households with fresh vegetables under the influence of rationalization and urbanization, 1850-1913]. PMID- 19678429 TI - [The new deontologic codes of Italian nurses: various considerations]. PMID- 19678430 TI - [The making of the Australian welfare system]. PMID- 19678432 TI - Diarrhea. Enterotoxigenic Bacteroides fragilis: newly recognized cause of inflammatory diarrhea. PMID- 19678431 TI - [Peasant migration from villages and social change in northern China in the republican period]. PMID- 19678433 TI - [Smoking cessation during pregnancy, newborn size, and NO-synthase activity in endothelial cells of the umbilical cord--secondary publication]. AB - Newborns of smokers had a lower weight, a smaller head circumference, and were shorter than newborns of smokers and ex-smokers, who stopped smoking early in pregnancy. The activity and concentration of the enzyme which produces the vasodilator nitric oxide were 36% and 47% lower, respectively, in endothelial cells of the umbilical vein from smokers compared with that from nonsmokers and ex-smokers. The findings suggest that smoking reduces nitric oxide production in the foetal vascular bed, contributing to retarded foetal growth caused by the reduction of vasodilatory capacity. PMID- 19678434 TI - [Fasting and nonfasting lipids--secondary publication]. AB - We tested the hypotheses that lipid levels change minimally in response to normal food intake and that nonfasting levels predict cardiovascular events. The maximum changes after normal food intake from fasting levels were as follows: total cholesterol -0.2 mmol/l, LDL cholesterol -0.2 mmol/l, HDL cholesterol -0.1 mmol/l, and for triglycerides +0.3 mmol/l. Highest versus lowest tertile of nonfasting total cholesterol, LDL cholesterol, and triglycerides, and lowest versus highest tertile of nonfasting HDL cholesterol predicted a 1.7-to 2.2-fold increased risk of cardiovascular events. PMID- 19678435 TI - [Nonfasting triglycerides and risk of ischemic stroke--secondary publication]. AB - The role of triglycerides in the risk of ischemic stroke remains controversial. We tested the hypothesis that increased levels of nonfasting triglycerides are associated with ischemic stroke in the general population. Men with a nonfasting triglyceride level 5 mmol/l had a multivariable, adjusted hazard ratio for ischemic stroke of 2.5 (95% confidence interval: 1.3-4.8) compared with men with a nonfasting triglyceride level < 1 mmol/l. The corresponding value in women was 3.8 (1.3-11). We conclude that the level of nonfasting triglycerides is associated with risk of ischemic stroke. PMID- 19678437 TI - Retraction. Improving quality and reducing inequities: a challenge in achieving best care. World Hosp Health Serv. 2008;44(3):16-31. PMID- 19678438 TI - Notice of retraction of abstracts. PMID- 19678436 TI - [The efficacy of the three available anti-tumour necrosis factor therapies in patients with rheumatoid arthritis. A meta-analytic literature review--secondary publication]. AB - Absolute treatment efficacy (via number needed to treat) of rheumatoid arthritis for each of the three available anti tumour necrosis factor alpha inhibiting therapies. Our aim was to indirectly compare the long-term efficacy of the available anti-tumour necrosis factor (TNF) therapies in patients with established rheumatoid arthritis. The results indicate that the absolute efficacy associated with use of these anti-TNF therapies is equivalent if applied in equivalent dosages. In standard dosages, infliximab (3 mg/kg/eight weeks) is only half as effective as adalimumab (40 mg/two weeks) and etanercept (25 mg twice weekly). PMID- 19678439 TI - [Role of psychiatrists in industrial medicine]. PMID- 19678440 TI - Two-year clinical evaluation of IPS Empress II ceramic onlays/inlays. AB - The stronger the ceramic material, the longer the restoration stays in the mouth. The current study evaluated the two-year clinical performance of a strong ceramic system, IPS Empress II, with increased strength on onlay/inlay restorations of molars. Teeth from 35 patients, including three premolars and 32 molars, were prepared for 28 onlay and seven inlay restorations with IPS Empress II ceramics. The restorations were cemented with a highly viscous, dual-curing luting composite cement (Bifix) and evaluated by two examiners using USPHS criteria at baseline (one week following insertion), six months, one year and two years. The baseline scores and recalls were assessed by Wilcoxon signed rank test. Statistically significant marginal discoloration at the Bravo level was found at the 12- and 24-month recalls (p=0.046). One debonding was statistically insignificant. No changes were observed with respect to anamnesis, such as any symptom from the TMJ or masticatory muscles. No restorations were replaced due to hypersensitivity or were missing at the two-year evaluation. Any wear on the restoration, antagonist tooth or any changes of proximal contacts were not observed. IPS Empress II Ceramics were found to be appropriate as onlay/inlay restorations for clinical use under the conditions of the current study. PMID- 19678441 TI - In vivo chlorhexidine stabilization of hybrid layers of an acetone-based dentin adhesive. AB - The current in vivo study evaluated the degradation of dentin hybrid layers in deep occlusal-surface resin composite restorations using TEM. Caries-free premolars scheduled for extraction as part of orthodontic treatment were prepared and restored, then extracted after 12 months. The adhesive used was a single bottle etch-and-rinse acetone-based product (Prime & Bond NT, Dentsply/Caulk). Control group restorations (n=8) were placed according to the manufacturer's instructions, while the experimental group received application of a 2% solution of chlorhexidine digluconate after etching and rinsing and prior to application of the adhesive. Extensive degradation was observed in all of the teeth in the control group after 12 months, while no degradation was observed in the experimental group. In vitro testing showed no significant difference in immediate microtensile bond strength between the control and experimental adhesive protocols. PMID- 19678442 TI - A 36-month clinical evaluation of ethanol/water and acetone-based etch-and-rinse adhesives in non-carious cervical lesions. AB - This double-blind randomized clinical trial compared different ethanol/water and acetone-based systems in non-carious cervical lesions over 36 months. MATERIALS AND METHODS: Eighty-four patients having at least one non-carious cervical lesion [NCCL] under occlusion were enrolled in this study. A total of 84 restorations were placed, half for each group (Adper Single Bond [SB] + FiltekA110 or One Step [OS] + MicroNew). All the materials were placed by two calibrated operators. Two other independent examiners evaluated the restorations at baseline, 6, 12, 18 and 36 months, according to slightly modified USPHS criteria. Statistical analysis between materials in each period was conducted using the Fisher's exact test (alpha=0.05), and performance of the materials in the baseline in comparison to each period was evaluated by McNemar's test (alpha=0.05). RESULTS: The 12-, 18- and 36-month retention rates for SB were 95.2% (12 and 18 months) and 92.3% (36 months). For OS, the retention rates were 83.3%, 73.8% and 56.4%, respectively, for each recall period. After 36 months, 10 OS restorations (25.7%) and seven SB restorations (17.9%) were rated as Bravo in the marginal discoloration item. CONCLUSIONS: The ethanol/water-based adhesive (Single Bond) that was evaluated showed a higher retention rate than the acetone-based system (One Step) after 36 months of clinical service. PMID- 19678443 TI - Dentin hypersensitivity: a randomized clinical comparison of three different agents in a short-term treatment period. AB - OBJECTIVE: Dentin hypersensitivity, or what patients may describe as "sensitive teeth," is defined as a short, sharp pain arising from exposed dentin in response to thermal, evaporative, tactile, electrical, osmotic or chemical stimuli. It is widely accepted that dentin hypersensitivity is an uncomfortable condition that also affects function and quality of life. This study determines the differences in efficiency of three desensitizing products when compared with a placebo. METHODS: A randomized controlled clinical trial was conducted to compare three different professional dentin desensitizer agents in 52 patients. The age and sex of the patients was recorded. Gluma Desensitizer (Heraeus Kulzer), UltraEZ (Ultradent Products, Inc) and Duraphat (Colgate Oral Pharmaceuticals, Inc, New York, NY, USA) were used as desensitizer agents and distilled water was used as the placebo. The baseline measurement of the dentin hypersensitivity was made by using a visual analog scale (VAS). Twenty-four hours and seven days after application of the desensitizer agents and placebo, a new VAS analysis was conducted for patients' sensitivity level. The desensitizer agents were compared in terms of mean values, and ANOVA was used for testing differences among the groups (p<0.05). RESULTS: The results showed that the mean pain scores of the placebo group were significantly higher than that of the study groups (p<0.05). The VAS analysis revealed a significant decrease in dentin hypersensitivity over time with the use of agents (p<0.05). No statistically significant difference was found among the three desensitizing agents (p>0.05). CONCLUSIONS: These three desensitizing agents, which contain different active ingredients, were effective in relieving dentin hypersensitivity. However, no superiority was found in dentin sensitivity relief among the agents. PMID- 19678444 TI - The effect of surface treatments on the micro-shear bond strength of a resin luting agent and four all-ceramic systems. AB - OBJECTIVE: The current study evaluated the micro-shear bond strength between a resin luting agent and four strengthened all-ceramic systems under different surface treatments. METHODS: Rectangular specimens of IPS Empress 2 (Ivoclar Vivadent), Cergogold (DeguDent), In Ceram Alumina (Vita) and Cercon (DeguDent) ceramics were fabricated and randomly divided into three groups: 1-no treatment; 2-etched with 9.5% hydrofluoric acid and 3-airborne-particle abraded with 50 microm aluminum oxide particles. The ceramic surfaces of the specimens were coated with a silane agent (Clearfil Porcelain Bond, Kuraray), then bonded with a resin-luting agent (Panavia F, Kuraray). A micro-shear bond test was carried out to measure the bond strength. Moreover, each ceramic surface was observed morphologically by scanning electron microscopy. The results were submitted to analysis of variance and Tukey's post-hoc analysis (p<0.05). RESULTS: The bond strength of all ceramic systems evaluated was affected by the surface treatments (p<0.05). The highest values for bond strength of IPS Empress 2 were found when the surface treatment used was hydrofluoric acid etching, followed by airborne particle abrasion treatment. On the other hand, airborne particle abrasion treatment and acid etching were not different for Cergogold and In Ceram Alumina ceramics, but they were higher when compared to the control (p<0.05). The highest bond strength to Cercon was found when it was treated with airborne particle abrasion with aluminum oxide. The SEM photographs showed that the hydrofluoric acid etching treatment affected the surface of IPS Empress 2 and Cergogold; however, Cercon and In Ceram surface morphology were not changed by the hydrofluoric acid etching. The airborne particle abrasion treatment altered the Cercon ceramic morphology but it did not change the other ceramic's surface. PMID- 19678446 TI - Bond strengths of resin cements to astringent-contaminated dentin. AB - The current study evaluated the micro-shear bond strength of two resin cements to astringent-contaminated dentin. Twelve occlusal dentin discs were prepared from extracted caries-free human molars and divided into two groups subjected to two types of resin cements, Panavia F (PF) and Variolink II (VL). Each disc was ground with 600 grit SiC paper and sectioned into two semi-disks, one for the normal dentin surface and the other for the contaminated dentin surface. For contaminated dentin, an astringent containing aluminum chloride was applied for two minutes and rinsed before the bonding procedures. A micro tygon tube was placed on the dentin surface following the bonding application and then filled with a resin cement. After the resin was polymerized, the specimen was kept in water for 24 hours before the micro-shear bond strengths evaluation. The micro morphology of the treated surfaces and resin-dentin interfaces were observed under a scanning electron microscope (SEM). Aluminum content under different dentin conditions was also examined. No significant differences were found between the dentin bond strengths to normal dentin and contaminated dentin surfaces in both the PF and VL groups (p>0.05). PF showed similar bond strengths to VL on normal and contaminated dentin (p>0.05). SEM observations of the VL groups revealed no differences in the treated dentin surfaces and the resin dentin interfaces between normal and contaminated dentin. However, for the PF group, an inconsistent etching pattern of the self-etching primer and gap formation at the interface of resin cement to contaminated dentin were observed. PMID- 19678445 TI - Influence of photoactivation protocol and light guide distance on conversion and microleakage of composite restorations. AB - PURPOSE: To evaluate the effect of light guide distance and the different photoactivation methods on the degree of conversion (DC) and microleakage of a composite. METHODS AND MATERIALS: Three photoactivation protocols (600 mW/cm2 x 40 seconds; 400 mW/cm2 x 60 seconds or 200 mW/cm2 x 20 seconds, followed by 500 mW/cm2 x 40 seconds) and three distances from the light source (0, 3 or 7 mm) were tested. Cylindrical specimens (5 mm diameter; 2 mm tall; n=3) were prepared for the DC test (FT-Raman). Class V cavities were made in 90 bovine incisors to conduct the microleakage test. The specimens were conditioned for 15 seconds with phosphoric acid (37%), followed by application of the adhesive system Prime & Bond NT (Dentsply/Caulk). The preparations were restored in bulk. The specimens were stored for 24 hours in distilled water (37 degrees C) before being submitted to the silver-nitrate microleakage protocol. The restorations were sectioned and analyzed under 25x magnification. RESULTS: Statistical analyses (two-way ANOVAs and Tukey test, alpha=0.05) found significance only for the factor distance (p=0.015) at the top of the composite for the DC test. Conversion was statistically lower for the 7 mm groups compared to the 0 and 3 mm groups, which were equivalent to each other. At the bottom of the specimens, none of the factors or interactions was significant (p<0.05). The Kruskal-Wallis test showed that, in general, the soft-start method led to lower microleakage scores when compared to the continuous modes, mainly when associated with a distancing of 7 mm (p<0.01). With the exception of specimens irradiated with 400 mW/cm2 that did not demonstrate variations on scores for the distances tested, higher microleakage was observed for shorter distances from the light source. CONCLUSIONS: Soft-start methods may reduce microleakage when the light guide distancing provides a low level of irradiance, which also causes a discrete reduction in the DC. PMID- 19678447 TI - In vitro wear of new indirect resin composites. AB - This in vitro study evaluated the toothbrush abrasion wear, three-body Alabama wear and two-body pin-on-disc wear of four commercial indirect resin composites. Enamel shades of Radica (R), Sculpture Plus (S), Belleglass-NG (B) and Gradia Indirect (G) were used. For measuring wear due to toothbrush abrasion, six specimens of each group were fabricated, then brushed in a toothbrush abrasion machine for 20,000 cycles. Material loss was determined by weighing and conversion to volume loss. Three-body wear was measured on six samples for each group using an Alabama-type wear testing machine for 400,000 cycles. Wear depth was measured with a contact profilometer. For two-body wear, five disc specimens were prepared and tested in a two-body wear-testing machine against hydroxypatite sliders for 25,000 cycles. Data were analyzed with one-way analysis of variance (ANOVA) and Tukey test (alpha=0.05). Wear was the highest in Sculpture Plus by all three methods tested and the lowest wear was observed in Belleglass-NG. No statistical difference in wear was noted from Radica. PMID- 19678448 TI - Fracture strength of endodontically-treated teeth restored with post and cores and composite cores only. AB - This study compared the fracture strength of different conditioned metallic posts, fiber-reinforced-composite posts and composite cores only in teeth without coronal tooth structure and determined failure modes after the fracture test. Post spaces were prepared in the root canals, and the teeth were randomly divided into seven experimental groups: Gr1: Titanium posts (ParaPost) + Silano-Pen (Bredent) + silane; Gr2: Titanium posts + 30 microm CoJet-Sand (3M ESPE) + silane; Gr3: Titanium posts + 50 microm Al2O3 + V-primer (Sun Medical); Gr4: Titanium posts + 50 microm Al2O3 + Alloy primer (Kuraray); Gr5: E-glass FRC post (EverStick); Gr6: Polyethylene fiber (Ribbond) + Resin impregnation and Gr7: Resin composite core only, with no posts. The posts were cemented using Panavia F 2.0 (Kuraray); coronal parts of the roots were etched, primed, bonded and composite cores were built-up. After thermocycling (5 degrees C-55 degrees C, 6000x), the fracture strength test was performed. The fracture strength of titanium posts (408 +/- 122 - 550 +/- 149 N) was significantly higher (p<0.05) than that of FRC posts (321 +/- 131 and 267 +/- 108 N for Everstick and Ribbond, respectively) or the group without posts (175 +/- 70 N) (Gr7) (ANOVA, Tukey's test). The group without posts resulted in complete core detachment (100%). In the E-glass FRC group, 60% adhesive core fracture occurred, covering >1/3 of the core and, in the polyethylene FRC group, 100% post-core detachment at the canal opening was observed. In all the titanium post applied groups (Gr1-Gr4), the posts remained in place with partial detachment of the core material from the post surface at varying degrees, depending on the conditioning method used. When no coronal tooth structure exist, the metal posts showed higher fracture strength values as opposed to the FRC post or no-post approach. PMID- 19678449 TI - Effect of ultrasonic versus manual cementation on the fracture strength of resin composite laminates. AB - This study evaluated the effect of conventional versus ultrasonic cementation techniques on the fracture strength of resin composite laminates. In addition, the failure modes were assessed. Window-type preparations 1 mm above the cemento enamel junction were made on intact human maxillary central incisors (N=60) of similar size with a depth cutting bur. All the prepared teeth were randomly assigned to six experimental groups (10/per group). Using a highly filled polymeric material (Estenia), laminates were produced and finished. The standard thickness of laminates in original tooth form was achieved using the impression molds made prior to tooth preparation. A three-step bonding procedure and dual polymerized resin composite cement (Panavia F 2.0) was employed. The cementation surfaces of the laminates were conditioned (CoJet-Sand, 30 microm SiO2) and silanized (ESPE-Sil). Laminates in Groups 1, 2, 3, 4 and 5 were cemented by five different operators under finger pressure and Group 6 was cemented ultrasonically (Amdent). After excess removal, the laminates were light polymerized. The specimens were stored in water at 37 degrees C for one month prior to the fracture test (universal testing machine, 1 mm/minute). Failure types were classified as: a) Cohesive failure within the composite laminate (Type A), b) Adhesive failure between the tooth and laminate (Type B) and c) Chipping of the laminate with enamel exposure (Type C). No significant difference was found among the mean fracture strength values of the laminates in all the experimental groups (ANOVA, p=0.251). The mean fracture strength values in descending order were: 513 +/- 197, 439 +/- 125, 423 +/- 163, 411 +/- 126, 390 +/- 94, 352 +/- 117 N for Groups 2, 5, 4, 3, 1 and 6, respectively. The majority of failure types was Type A (30/60). While Type B failure was not observed in Group 6 (0/10), Group 1 presented a more frequent incidence of this failure (6/10). The two cementation techniques did not effect the fracture strength of composite laminates, but failure types varied between groups, being more favorable for the ultrasonically cemented group. PMID- 19678450 TI - An in vitro comparison of different cementation strategies on the pull-out strength of a glass fiber post. AB - PURPOSE: To evaluate the effect of different strategies for post cementation on the pullout bond strength of a double-tapered glass fiber post cemented into a root canal. MATERIALS AND METHODS: The root canals of 70 single-rooted bovine teeth (16 mm-length) were prepared to 9 mm using the preparation drill of a double-tapered glass fiber post system (White Post DC, FGM). Each specimen was embedded in a plastic cylinder using acrylic resin up to 3 mm of the most coronal portion of the specimen and allocated into one of seven groups (n=10) based on strategies for cementation: Gr1-ScotchBond Multi Purpose plus (SBMP) + Relyx ARC resin cement; Gr2-Single Bond + Relyx ARC; Gr3-ED Primer + Panavia F resin cement; Gr4-SBMP + AllCem resin cement; Gr5-Relyx ARC; Gr6-Relyx Unicem resin cement; Gr7-Relyx Luting 2 glass ionomer cement. After cementation, the specimens were stored for seven days (in a humid environment at 37 degrees C) and submitted to pullout bond strength testing (the inferior part of each specimen was fixed and the fiber post was pulled out). The data (Kgf) were submitted to statistical analysis (one-way ANOVA and post-hoc Tukey tests, alpha=.05). The tested specimens were analyzed under the microscope and SEM for fracture analysis. RESULTS: The strategy for post cementation affected the pullout retentive strength (Kgf) (p<0.0001) significantly. Gr6 (37.7 +/- 8a), Gr1 (37.4 +/- 5.7a) and Gr4 (31.6 +/- 6.6ab) presented the highest pullout bond strengths. Gr2 (12.2 +/- 5.6c), Gr3 (6.5 +/- 5.2c) and Gr7 (5.1 +/- 2.8c) presented the lowest pullout bond strengths. Gr5 (24.2 +/- 7.4b) was similar to Gr4 and inferior to Gr6 and Gr1. CONCLUSION: The use of a three-step etch-&-rinse adhesive system appears to be effective. The application of other adhesive systems (single-bottle etch-& rinse and self-etch adhesive systems) did not present high pullout strength values. The simplified self-adhesive resin cement (without adhesive application) presented good retentive performance. Further studies should be conducted. PMID- 19678451 TI - Effect of the C-factor and dentin preparation method in the bond strength of a mild self-etch adhesive. AB - This study evaluated the effect of the C-factor and dentin preparation method (DPM) in the bond strength (BS) of a mild self-etch adhesive; the study also observed the SEM superficial aspects of the corresponding smear layer. For purposes of this study, 25 molars (n=5) were used in a bond strength test. The molars were divided into two parts (buccal and lingual): one part received a Class V cavity (C-factor=3) and the other received a flat surface (C-factor=0) with the same bur type (coarse diamond or carbide bur and fine diamond or carbide bur), both within the same dentin depth. Five teeth were prepared with wet 60 grit and 600-grit SiC papers. After restoration with Clearfil SE Bond, microtensile beans (0.8 mm2) were prepared and tested after 24 hours in a universal testing machine (0.5 mm/minute). An additional two teeth for each DPM were prepared for SEM evaluation of the smear layer superficial aspects. The BS values were submitted to one-way ANOVA, considering only the DPM (flat surfaces) and two-way ANOVA (C-Factor x DPM, considering only burs) with p=0.05. Although the DPM in the flat surfaces was not significant, the standard deviations of carbide bur-prepared specimens were markedly lower. The BS was significantly lower in cavities. The fine carbide bur presented the most favorable smear layer aspect. It was concluded that different dentin preparation methods could not prevent the adverse effect in bond strength of a high C-factor. A coarse cut carbide bur should be avoided prior to a mild self-etch adhesive, because it adversely affected bond strength. In contrast, a fine cut carbide bur provided the best combination: high bond strength with low variability, which suggests a more reliable bond strength performance. PMID- 19678452 TI - Effect of Er,Cr:YSGG laser on the microtensile bond strength of two different adhesives to the sound and caries-affected dentin. AB - OBJECTIVE: This study examined the effect of Er,Cr:YSGG laser irradiation on the microtensile bond strength (microTBS) of a three-step etch-and-rinse and a two step self-etch adhesive to sound and caries-affected dentin. METHODS: Sixteen freshly extracted human molars with occlusal dentin caries were used. The caries lesion was removed by one of the following methods: conventional treatment with burs or Er,Cr:YSGG laser (Waterlase MD, Biolase). The adhesive systems (AdheSE, Ivoclar Vivadent and Scotchbond Multi Purpose, 3M ESPE) were applied to the entire tooth surface according to the manufacturers' instructions. Resin composites were applied to the adhesive-treated dentin surfaces and light-cured. Each tooth was sectioned into multiple beams with the "non-trimming" version of the microtensile test. The specimens were subjected to microtensile forces (BISCO Microtensile Tester, BISCO). The data was analyzed by three-way ANOVA and independent t-tests (p=0.05). RESULTS: Er,Cr:YSGG laser irradiation exhibited similar microTBS values compared to that of conventional bur treatment, regardless of the adhesive system and type of treated dentin. The self-etch system revealed lower microTBS values, both with conventional and laser treatment techniques, compared to the etch-and-rinse adhesive in sound and caries-affected dentin (p<0.05). CONCLUSION: Er,Cr:YSGG laser irradiation did not negatively affect the bonding performance of adhesive systems to sound and caries-affected dentin. PMID- 19678453 TI - Shear bond strength evaluation of resin composite bonded to GIC using three different adhesives. AB - The current study evaluated the bonding ability of composite to glass ionomer cement (GIC) using three different bonding systems. One hundred samples of composites bonded to GIC were prepared and divided into five groups. In Group A, the composite was bonded to GIC after the initial setting of the GIC being employed as a total-etch adhesive. In Group B, the self-etch primer was employed to bond composite to GIC before the initial setting of the GIC. In Group C, the self-etch primer was employed to bond composite to the GIC after the initial setting of the GIC. In Group D, the GIC-based adhesive was employed to bond composite to the GIC before the initial setting of the GIC. In Group E, the GIC based adhesive was employed to bond composite to the GIC after the initial setting of the GIC. Shear bond strength analysis was performed at a crosshead speed of 0.5 mm/minute. The results were tabulated and the statistical analysis was performed with one-way ANOVA; the Tukey's test showed that the bond strength of composite to GIC was significantly higher for the self-etch primer group employed on unset GIC and the GIC-based adhesive group employed on the set GIC for bonding composite to GIC. PMID- 19678454 TI - Effects of storage temperature on the shelf life of one-step and two-step self etch adhesives. AB - Recently, self-etch adhesive systems, the one-step (all-in-one) and two-step adhesive systems, have become widely utilized due to their simplified application procedures and low technique sensitivity. In the current study, in order to understand the effects on shelf life of the two types of self-etch adhesives, the effects from storage temperature and time period on the alteration stage of 10 methacryloxydecyl dihydrogen phosphate (MDP)-based, a one-step adhesive (all-in one adhesive system) and an MDP-based self-etching primer (two-step adhesive system) were examined. Clearfil Tri-S Bond (TSB), an MDP-based one-step adhesive (all-in-one adhesive system), and Clearfil Mega Bond Primer (MBP), an MDP-based self-etching primer (two-step adhesive system), were used. Both TSB and MBP, received within two days after they were produced, were immediately stored at 8 degrees C, 20 degrees C or 40 degrees C for 1, 3, 7 and 14 weeks, respectively. At the end of each storage period, 13C NMR observations were performed by an EX 270 spectrometer. In addition, NMR observations of TSB and MBP were immediately performed within two days, after both were produced as a control. The effects from the storage temperature and time period on the alteration rate and stage of TSB and MBP were examined by determining the amount of hydrolyzed 2-hydroxyethyl methacrylate (HEMA) and MDP in both materials. Specimens bonded to dentin by using TSB or MBP in different alteration stages were prepared. The degradation stage effect of TSB or MBP on bond durability was examined by measuring the shear bond strength before and after thermocycling (20,000 times). With increases in storage temperature and time period, the relative intensities of the NMR peak "epsilon" assigned to both methylene carbons in the ethylene glycol (EG) and to the NMR peak "zeta" assigned to the terminal methylene carbon bonded to the hydroxy group in the 10-hydroxydecyl dihydrogen phosphate (HDP) produced by the hydrolysis of the ester portion in HEMA or MDP, respectively, increased. The alteration stages of TSB and MBP were strongly dependent on storage temperature and time period. When TSB or MBP, stored at 40 degrees C for 14 weeks, was applied to dentin, specific decreases in mean bond strength were observed in both adhesive systems. However, the application of thermocycling did not exhibit any specific decreases in the mean bond strength, even though the alteration stage of TSB and MBP progressed. From the results of the current study, storage temperature and time period significantly affect the alteration rate and stage of TSB and MBP. However, TSB and MBP exhibit expectant bond strength and bond durability when both are stored below 20 degrees C. PMID- 19678455 TI - Chlorhexidine-containing acid conditioner preserves the longevity of resin-dentin bonds. AB - The current study evaluated the effect of 2% chlorhexidine digluconate (CHX) on the immediate and six-month resin-dentin bond strength (BS) and nanoleakage pattern (NL) of etch-and-rinse adhesives when applied in aqueous or associated to the phosphoric acid conditioner. The occlusal enamel of 42 caries-free extracted molars was removed in order to expose a flat dentin surface. In groups 1 and 2 (control-C), the surfaces were acid etched with conventional phosphoric acid, and the adhesives Prime&Bond NT (PB) and Adper Single Bond 2 (SB) were applied after rinsing, drying and rewetting with water. In groups 3 and 4 (Ac/CHX), the adhesives were applied in a similar manner, however, a 2% CHX-containing acid was previously applied. In groups 5 and 6 (CHX), the adhesives were applied according to the control group; however, the rewetting procedure was performed with an aqueous solution of 2% CHX for 60 seconds. Composite buildups (Opallis, FGM) were constructed incrementally, and the specimens were longitudinally sectioned in the "x" and "y" directions to obtain bonded sticks (0.8 mm2) to be tested in tension at 0.5 mm/minute immediately or after six months of water storage. For NL, two bonded sticks from each tooth were coated with nail varnish, placed in silver nitrate and polished down with SiC paper. Resin-dentin interfaces were analyzed by EDX-SEM. The BS and NL data from each adhesive was submitted to two-way repeated measures ANOVA and Tukey's test (alpha=0.05). After six months of water storage, significant reductions in BS were observed for both adhesives in the control group (p<0.05). When Ac/CHX or CHX was used, no significant reductions in BS were observed for both systems. Nanoleakage was more evident in the control group than in the experimental groups (p<0.05), even after six months. The use of CHX in an aqueous solution or associated with the acid conditioner was effective for reducing degradation of resin-dentin bonds after six months of water storage. PMID- 19678456 TI - Utilization of occlusal index and layering technique in class I silorane-based composite restorations. PMID- 19678457 TI - The combination of a mineral trioxide aggregate and an adhesive restorative approach to treat a crown-root fracture coupled with lateral root perforation in a mandibular second molar: a case report. AB - The current paper describes a modified treatment procedure for a traumatized mandibular left second molar resulting in a crown-root fracture and root perforation with the fracture line below the gingival attachment and alveolar bone crest. After the mobile crown-root fragment was extracted, the root perforation was obturated with mineral trioxide aggregate (MTA), and the subgingival defect was directly repaired with polyacid-modified resin composites (Ionosite Baseline). A 24-month recall showed no evidence of periodontal inflammation and no adverse symptoms, and the treated tooth exhibited good healing and normal function. PMID- 19678458 TI - Where are the falconers? A meditation on leadership. PMID- 19678459 TI - Living with advanced cancer and short life expectancy: patients' experiences with managing medication. AB - Our aim is to explore patients' experiences of using medicines when they are living with far-advanced cancer and short life expectancy; our method is a qualitative interview study. At a daycare centre at a palliative clinic in Norway, we interviewed 15 patients with advanced incurable cancer with multiple metastases who had a short life expectancy. We found that in taking their medications, they feared losing control, becoming addicted, or suffering harmful effects. Non-compliance was the rule, not the exception: patients juggled doses or dosage intervals, or they stopped taking the medications. They wanted to take as little medication as possible and self-manage it to gain control over their lives. We concluded that patients need to discuss their medication practice. If they choose alternative medication strategies, that choice must be respected. For patients, the issue is self-management, not compliance. Patients with a short life expectancy want to negotiate their medication practice with health care professionals and take an active role in tailoring it to suit their preferences. Health professionals should therefore consider a concordance rather than a compliance model for these patients. PMID- 19678460 TI - Referrals of cancer versus non-cancer patients to a palliative care consult team: do they differ? AB - This retrospective study compared 100 consecutive non-cancer (NC) patients referred to a palliative care consult team (PCT) in a Swiss university hospital to 506 cancer (C) patients referred during the same period. The frequencies of reported symptoms were similar in both groups. The main reasons for referral in the NC group were symptom control, global evaluation, and assistance with discharge. Requests for symptom control predominated in the C group. Prior to the first visit, 50% of NC patients were on opioids, compared to 58% of C patients. After the first visit, the proportion of NC patients on opioids increased to 64% and the proportion of C patients to 73%. The median daily oral morphine equivalent dose for NC patients taking opioids prior to the first PCT visit was higher than that for C patients (60 mg versus 45 mg). At the time of death or discharge, the percentage of NC patients on opioids was 64%, while that of C patients was 76%. Moreover, NC patients were on significantly lower median doses of opioids than C patients (31 mg versus 60 mg). Over half the NC patients died during hospitalization, as compared to 33% of C patients. Only 6% of NC patients were discharged to palliative care units, as compared to 22% of C patients. PMID- 19678462 TI - Lessons in death and dying. PMID- 19678463 TI - Using philosophy to help manage the fear of death. PMID- 19678461 TI - Might massage or guided meditation provide "means to a better end"? Primary outcomes from an efficacy trial with patients at the end of life. AB - This article reports findings from a randomized controlled trial of massage and guided meditation with patients at the end of life. Using data from 167 randomized patients, the authors considered patient outcomes through 10 weeks post-enrollment, as well as next-of-kin ratings of the quality of the final week of life for 106 patients who died during study participation. Multiple regression models demonstrated no significant treatment effects of either massage or guided meditation, delivered up to twice a week, when compared with outcomes of an active control group that received visits from hospice-trained volunteers on a schedule similar to that of the active treatment arms. The authors discuss the implications of their findings for integration of these complementary and alternative medicine therapies into standard hospice care. PMID- 19678464 TI - Moving culture beyond ethnicity: examining dying in hospital through a cultural lens. PMID- 19678465 TI - Enhancing interprofessional education in end-of-life care: an interdisciplinary exploration of death and dying in literature. PMID- 19678466 TI - Innovative pediatric palliative care programs in four countries. AB - Over the past three decades, pediatric palliative care programs have been developed and refined throughout the world. The purpose of this study was to provide information on experiences from four of those programs, yet we acknowledge that there are many other innovative programs that deserve recognition for the services they provide to children and families. This study is limited in that it is unable to compare outcomes from the four programs, such as patient-reported quality of life, that might help to better understand the impact of pediatric palliative care. Nonetheless, information sharing can inspire and educate others with the overarching goal of globally advancing pediatric palliative care. PMID- 19678467 TI - East meets West. Spirituality training in palliative care: a Taiwanese-Australian partnership. PMID- 19678468 TI - Treatment of medication intolerance with lactase in a complex palliative care patient. PMID- 19678469 TI - A final toast. PMID- 19678470 TI - Childhood sexual abuse, PTSD, and borderline personality disorder. Understanding the connections. PMID- 19678471 TI - Youth and disordered eating. PMID- 19678473 TI - Drugs to treat depression. PMID- 19678472 TI - Adolescent depression: assessment and treatment. AB - Recent recommendations include screening for depression in adolescents when systems for diagnosis, treatment, and follow up are in place; conducting a risk assessment of factors such as parental depression, comorbid mental health or chronic medical conditions, and having had a major negative life event; and that intervention with selective serotonin reuptake inhibitor drugs when adequate monitoring is available, psychotherapy, and combined treatment have been found to be effective (Agency for Healthcare Research and Quality, 2009). Kayla benefited from this model of early detection through the school nurse and early intervention and satisfactory monitoring of treatment and safety. PMID- 19678474 TI - Understanding chirality and stereochemistry: three-dimensional psychopharmacology. AB - Drugs and the places in the body they bind to are each three-dimensional molecular structures. Stereoisomers are compounds that possess the same molecular and structural formula but differ in their three-dimensional configuration. Chiral compounds have two mirror-image stereoisomer forms called enantiomers. Compounds that contain both mirror-image enantiomers in equal proportions are referred to as racemic mixtures or racemates. The presence of a second enantiomer in racemic mixtures can be associated with various problems. The second enantiomer may be less biologically active but could interfere with the more biologically active enantiomer by binding to the same site. Adverse reactions can sometimes be attributed to one of the enantiomers. Each enantiomer may bind to metabolic enzymes differently, resulting in different rates of metabolism and clearance. They may interact differently with other drugs. The investigation, development, and use of stereochemically pure drugs have been encouraged by the U.S. Food and Drug Administration to improve the efficacy, tolerability, and safety of marketed drugs. PMID- 19678475 TI - Distinguishing between personality disorders, stereotypes, and eccentricities in older adults. AB - Personality disorders, especially in older adults, are among the most difficult psychiatric disorders for nurses to assess. When aging further complicates these disorders, nurses' therapeutic skills are challenged. It has long been thought that personality disorders "age out," but new research indicates that personality disorders may in fact continue throughout the life span. In addition, the primary and secondary changes of aging further complicate assessment. Assessment of personality disorders in older adults may also be distorted by ageist stereotypes and a lack of understanding of cultural context. Likewise, nurses must be careful about misinterpreting "eccentric" older adult behavior as a personality disorder. In this article, we focus on assessment challenges in older adults to help nurses distinguish between characteristics of personality disorders, stereotypes, and eccentricities in this population. PMID- 19678476 TI - Loss of a companion animal: understanding and helping the bereaved. AB - Pet owners may face numerous animal losses in a lifetime. Grief following pet loss is often misunderstood and devalued. Nurses are likely to encounter patients and families mourning the loss of a companion animal. This article outlines the grief process and offers practical suggestions for assisting those who are bereaved. PMID- 19678477 TI - Computer-assisted CBT for depression & anxiety: increasing accessibility to evidence-based mental health treatment. AB - Cognitive-behavioral therapy (CBT) is the most effective nonpharmacological treatment for almost all mental disorders, especially anxiety and depression. The treatment is time limited, encourages self-help skills, is problem focused, is inductive, and requires that individuals develop and practice skills in their own environment through homework. However, most of those with mental health issues are unable to seek help because of factors related to treatment availability, accessibility, and cost. CBT is well suited to computerization and is easy to teach to nurses. In this article we describe outcome studies of computer-assisted CBT (cCBT), outline the current technologies available, discuss concerns and resistance associated with computerized therapy, and consider the role of nurses in using cCBT. PMID- 19678478 TI - Consumer-operated self centers: environment, empowerment, and satisfaction. AB - Consumer-operated self-help centers were designed to provide social environments that promote participant empowerment and satisfaction. This exploratory, descriptive study examined how variance in empowerment and satisfaction scores could be explained by participants' perceptions of the social environment factors (relationship, personal growth, and systems maintenance and change) and quantity of participation. Participants (N = 144) involved in consumer-operated self-help centers completed a four-part, 161-item survey designed to capture perceptions of satisfaction, empowerment, social environment factors, quantity of center participation, and demographic data. Significant relationships were found between participant satisfaction and the three social environment factors. Findings also indicated that participant empowerment was related to quantity of self-help center involvement. From these exploratory analyses, recommendations are made on how to improve consumer-run self-help center operations. PMID- 19678479 TI - Aged care nursing lament. PMID- 19678481 TI - Free trade agreements and health services. PMID- 19678482 TI - Enrolled nurses: towards a level playing field. PMID- 19678483 TI - 'Doctor/Auntie' Sister Carmel--a tribute to a legendary paediatric nurse. PMID- 19678484 TI - Health care disaster ethics: a call to action. PMID- 19678485 TI - Superannuation update for nurses. PMID- 19678486 TI - Are we there yet? PMID- 19678487 TI - Nursing patients with Creutzfeldt-Jakob disease. Are you at risk? PMID- 19678488 TI - Jack of all trades in the operating theatre. PMID- 19678489 TI - Sharing primary health expertise with Aboriginal health workers. PMID- 19678490 TI - Studying the transition to maternal child health nurse. PMID- 19678491 TI - After hours palliative care. PMID- 19678492 TI - Demanding times ahead. PMID- 19678493 TI - [HPV and effectiveness of chlamydia control]. AB - Liberalization of sexual behaviour has brought about an increased outcome of the most common sexually transmitted diseases in people under 25 years, such as infections caused by chlamydia and carcinogenic human papillomaviruses (HPV) as well as their severe complications such as infertility and cancers of the anogenital region in those at the reproductive age. HPV vaccination and screenings are new possibilities for the control of these infections and their consequences. The herd immunity raised by vaccinating can protect as many as 25% of non-vaccinees as well. Screenings of birth cohorts of vaccinees aim to break the transmission chain of chlamydia. PMID- 19678494 TI - [How effectively should hyperglycemia be treated in a type 2 diabetes patient?]. AB - Hyperglycemia involves an increased risk of macrovascular and microvascular diseases. Despite convincing epidemiologic evidence, research results of the efficacy of treatment of hyperglycemia on cardiovascular events are conflicting. The results published in 2008 of large studies on the treatment of hyperglycemia have caused debate. According to the ACCORD study, intensive treatment of hyperglycemia can be deleterious in high-risk patients with a long history of the disease. Long-term results of the UKPDS study speak for intensive care at the early stage. A comprehensive and early medical care will yield the best long-term results in the treatment of type 2 diabetes. PMID- 19678495 TI - [Skin contact reactions caused by plants]. AB - Plants and plant products cause various contact reactions to the skin and mucosae. The skin reaction is usually dermatitis, such as irritant contact dermatitis, allergic dermatitis and a dermatitis caused by an airborne allergen. Other symptom pictures include allergic and non-allergic contact urticaria, protein contact dermatitis and phytophotodermatitis. Actinic reticuloid, usually induced by contact allergy, can also be considered a contact reaction. PMID- 19678496 TI - [Burn injuries caused by sauna air]. AB - Burn injuries caused by hot air sauna burns constitute a Finnish burn rarity. The patients are usually middle-aged men having passed out on the sauna benches under the influence of alcohol. Sauna air causes a deep injury penetrating all layers of the skin, accompanied with necrosis of the subcutaneous tissue and consequent rhabdomyolysis. The initially harmless-looking erythema of the skin rapidly transforms into a third-degree burn. Therapy includes the prevention of kidney damage and surgery. Local flaps are recommended for the treatment of tissue defects caused by destruction of deep tissues and amputations. PMID- 19678497 TI - [Surgical site infections at Toolo hospital and the dog days myth]. AB - INTRODUCTION: In the Finnish folklore, the time from June 23 to August 23 has been called "rotten month" (dog days), whereupon wound healing is delayed due to infections. PATIENTS AND METHODS: A retrospective comparison was made of surgical infections entered in the infections register at Toolo hospital over the dog days of 2002 to 2005. RESULTS: Of 49517 surgical operations, 8 % were performed during dog days, during which the number of contaminated and infected surgeries was higher. CONCLUSIONS: The risk of deep surgery wound infection of organ infection during dog days is two times higher than at other times. PMID- 19678498 TI - [Treatment of sudden deafness]. AB - Sudden sensorineural hearing loss can be accompanied by sensation of aural fullness or pressure, and tinnitus. External auditory canal, middle ear and neurological status are normal. Hearing of speech and whispered voice are impaired, and tuning fork tests show a sensorineural hearing loss. Decreased air and bone conduction hearing thresholds are observed in pure tone audiometry. An urgent consultation of an otolaryngologist is indicated. Systemic corticosteroids are usually applied as a brief course. Follow-up audiogram is necessary. PMID- 19678499 TI - [No thyroxin dose seemed sufficient--why?]. AB - The health status of a woman with type 1 diabetes was followed during pregnancy in a maternity hospital. In addition to insulin, thyroxine therapy was applied due to hypothyreosis as a consequence of Basedow disease. Nephropathy and significant proteinuria had developed as complications of diabetes, with the proteinuria increasing considerably during pregnancy. Simultaneously thyroxine doses increased to exceptionally high levels. The cause of increasing doses was found to be secretion of thyroxine into urine. The phenomenon has been described earlier, but it is not generally recognized. PMID- 19678500 TI - Sentinel lymphadenectomy for cutaneous melanoma: who, what, and why. AB - Knowledge of the metastastic status of the regional nodes provides important prognostic information for patients with cutaneous melanoma. Sentinel lymph node (SLN) biopsy has become the standard method for staging the regional nodes of patients with melanoma. The decision algorithm for using SLN biopsy is based upon the primary tumor depth, and utilization of SLN biopsy can spare patients the potential morbidity associated with complete lymph node dissection. Current studies are investigating the possibility of further limiting the need for complete lymph node dissections in patients with positive sentinel nodes. PMID- 19678501 TI - Blastomycosis: case report investigating a persistent pulmonary lesion in an immunocompromised patient. AB - Blastomycosis is a well known infection caused by Blastomyces dermatitidis. It appears usually as a mild and self-limited disease, but disseminated cases are seen, especially in immunocompromised patients. In organ transplant recipients, fungal infections play an important role and in some cases can be fatal, but blastomycosis is a rare and uncommon condition in this setting and specifically in renal transplant patients. Its occurrence is probably due to previous exposure, but sometimes there is no clear history that indicates such exposure. We describe a patient who underwent a renal transplant and developed pulmonary blastomycosis. PMID- 19678502 TI - Exercise and weight control. PMID- 19678503 TI - Silent aspiration: results of 2,000 video fluoroscopic evaluations. AB - The purpose of this retrospective study of aspiration and the lack of a protective cough reflex at the vocal folds (silent aspiration) was to increase the awareness of nursing staffs of the diagnostic pathology groups associated with silent aspiration. Of the 2,000 patients evaluated in this study, 51% aspirated on the video fluoroscopic evaluation. Of the patients who aspirated, 55% had no protective cough reflex (silent aspiration). The diagnostic pathology groups with the highest rates of silent aspiration were brain cancer, brainstem stroke, head-neck cancer, pneumonia, dementia/Alzheimer, chronic obstructive lung disease, seizures, myocardial infarcts, neurodegenerative pathologies, right hemisphere stroke, closed head injury, and left hemisphere stroke. It is of high concern that the diagnostic groups identified in this research as having the highest risk of silent aspiration be viewed as "red-flag" patients by the nursing staff caring for them. Early nursing dysphagia screens, with close attention to the clinical symptoms associated with silent aspiration, and early referral for formal dysphagia evaluation are stressed. PMID- 19678504 TI - Hemolytic uremic syndrome: a case review. AB - Hemolytic uremic syndrome is a complex disease that impacts multiple body systems. Knowledge gained from cases has increased understanding of etiologic factors, presenting symptoms, diagnostic laboratory findings, and the disease process. In rare cases, severe neurological symptoms are evident. This 20-year old woman presented with bloody diarrhea that progressed quickly to respiratory distress requiring intubation and to the development of status epilepticus, controlled only by a barbiturate coma. Ongoing nursing care in the critical care unit was vital in preventing complications and promoting a positive outcome. PMID- 19678505 TI - The effectiveness of multistrategies on disruptive vocalization of people with dementia in institutions: a multicentered observational study. AB - The aim of this study was to evaluate the effectiveness of the daily interventions used by the nurses on disruptive vocalization (DV). DV includes all types of disturbing or unacceptable vocal expression: repetitive vocalization, verbal or nonverbal utterances, presented as inappropriate language, repeated and insistent demands, repeated calling out, shouting, complaining, or moaning that does not pertain to their circumstances or environment. A convenience sample of five nursing homes from the north of Italy, in the Friuli Venezia Giulia region, was included in the study. A randomized selection of 87 daily shifts was selected. Institutionalized patients with dementia, but with no associated psychiatric disorders, were eligible. Nurses involved in the study added patients progressively. Nurses involved were asked to keep diaries to record strategies and durations for each episode of DV encountered during the allotted shift. In the total amount of observation time (36,540 minutes), 23.6% (8,653 minutes) of nursing care time involved working with and managing DV patients. The nurses recorded an average of 6.5 (302/46) vocalizations on morning shifts and 7.3 (302/41) during afternoon shifts, with an average duration of about a quarter of an hour each. Managing DV with multistrategies reduces the duration of the DV episode and increases the perceived effectiveness of management. PMID- 19678506 TI - Epilepsy patients' conceptions of epilepsy as a phenomenon. AB - This study addressed epilepsy patients' conceptions of epilepsy as a phenomenon and emotions related to those conceptions. Nineteen outpatients were interviewed, and data were analyzed according to the phenomenographical methodology. Patients described epilepsy in six qualitatively different ways: Epilepsy is (a) an illness related to physical disturbances, (b) a condition related to physical disturbances, (c) a mental disturbance related to lack of mental capacity, (d) a handicap related to psychological and/or social aspects, (e) an identity related to being an epileptic, and (f) a punishment. The emotions confidence, happiness, hope, and annoyance were related to epilepsy as an illness or a condition, whereas shame, fear, sorrow, and guilt were related to the other four categories. This study indicated that, to patients, the phenomenon of epilepsy is above all a psychosocial nature and in that dimension closely related to negative emotions. PMID- 19678507 TI - The surgical treatment of trigeminal neuralgia: overview and experience at the University of Florida. AB - Trigeminal neuralgia, also called tic douloureux, is a commonly misdiagnosed disorder characterized by intense facial pain. It is a chronic pain disorder that affects the fifth cranial nerve, usually in the fifth to seventh decade of life. Medication is the first line of treatment but frequently fails over time. At that point, many patients seek surgical intervention. This study reviews 108 patients treated over a 1-year period at the University of Florida with one of two surgical procedures: radiofrequency lesioning and microvascular decompression. The short-term results of this experience are reviewed and discussed. PMID- 19678508 TI - Software-assisted spine registered nurse care coordination and patient triage- one organization's approach. AB - Back disorders encompass a spectrum of conditions, from those of acute onset and short duration to lifelong disorders. The use of a traditional spine center model of patient flow, in which the patient is scheduled the first available appointment without an initial assessment of spine-related symptoms at West Virginia University Spine Center, Morgantown, West Virginia, resulted in frustration and delays for the spine patient and referring physician dissatisfaction. Today, the use of a software-assisted spine patient triage and registered nurse care coordinator patient navigation system in this multidiscipline, multimodality comprehensive spine program provides quick and efficient patient triage to the appropriate level of spine care (surgeon vs. nonsurgeon). The model consists of five major steps, which are explored in this article: medical history intake; films or studies retrieval; rapid review of the patient's medical condition and diagnostics by a spine specialist preappointment and subsequent triage to the appropriate level of spine care; registered nurse care coordinator patient education and guided navigation through the patient's preferred treatment plan; and last, diagnostic study, pain injection, and provider scheduling. Patient satisfaction scores, referring physician satisfaction scores, and resultant impact on referral volumes, ancillary utilization, workload productivity, and surgical yield demonstrate that this new approach to patient triage has made significant improvements in efficiency, productivity, and service. PMID- 19678509 TI - Caregivers' experience: bimonthly participation in a year-long research interview process. AB - New caregivers of stroke survivors participated in a bimonthly research interview process for a year-long intervention study. The purpose of this current project was to describe the telephone interview experience for those in the study's control group. We purposively selected 14 participants to answer a telephone survey. The following themes emerged: (a) looking forward to talking with someone, (b) feeling helped and staying connected, (c) being busy caregiving and taking care of business, (d) helping others, and (e) being ambivalent or negative about the interview process. Orem's self-care deficit nursing theory was then applied to reflect upon these themes. It is important for nurse researchers to remember that interviews used for data collection are not neutral or noninteractive. PMID- 19678510 TI - Investing in the future. PMID- 19678511 TI - Why patient safety must be at the top of the agenda at every level. PMID- 19678512 TI - Action on intolerance. AB - A research programme involving four practice nurses promoting the benefits of healthy eating and identifying patients with food intolerance has paid dividends. PMID- 19678513 TI - Seeing off obstacles. AB - There is a good deal more to making innovation happen than removing barriers and providing funding. Meticulous planning is key. PMID- 19678514 TI - Our driving passion. AB - Nurses are key members of the medical team that works at racing events including the Silverstone Grand Prix. PMID- 19678515 TI - A promise that has to be kept. PMID- 19678516 TI - A framework for mentor support in community-based placements. AB - This article explores the development and provision of a framework for supporting mentors working in community settings. A working group was set up to explore the provision of mentor support in community placements in Leicester, Leicestershire and Rutland, which was restructured to meet the demands of Nursing and Midwifery Council standards and to demonstrate a commitment to supporting mentors. This was done by providing regular mentoring update sessions, a regular newsletter, visiting lecturer contact and practice-held mentor resource packs. Evaluation of the update sessions was generally positive; the main criticism was the lack of pre-course information, which has since been addressed. PMID- 19678518 TI - Pulmonary embolism. PMID- 19678517 TI - An overview of meningitis and meningococcal septicaemia. AB - This article provides an overview of meningitis and meningococcal septicaemia, which can have devastating effects. Nurses working in acute and primary care need to be able to recognise the causes and symptoms of these conditions, and have up to-date knowledge of treatment, prevention and potential after-effects. PMID- 19678519 TI - Assessment and management of patients with wound-related pain. AB - Wound-related pain can cause significant distress and may lead to reduced quality of life. This article provides an overview of the management and treatment of pain in patients with chronic wounds. The importance of undertaking a thorough pain assessment using pain measuring scales is also discussed. PMID- 19678521 TI - The learning divide. PMID- 19678520 TI - Reswick and Rogers pressure-time curve for pressure ulcer risk. Part 1. AB - Pressure ulcers are one of the most potentially devastating complications in individuals confined to a bed or a wheelchair for an extended period. Severe pressure ulcers may form in deep tissues overlying bony prominences, and only at a later stage become visible, or may even induce a full-thickness breakdown of the soft tissues at the affected site. A new type of pressure ulcer known as'deep tissue injury' has therefore been defined internationally. To understand the aetiology of deep tissue injury, health professionals should be able to predict whether or not a certain state of internal mechanical loads in deep tissues, such as tissue deformations and forces per unit area of tissue, would lead to localised irreversible cell damage. Part one of this article explains the concepts of injury thresholds as related to deep tissue injury. Some serious flaws in the classical, commonly used Reswick and Rogers pressure-time curve are analysed, and an alternative contemporary tissue injury threshold, the sigmoid threshold, is suggested. Part two of this article describes recent and ongoing work aimed at defining injury thresholds that are specific for deep tissue injury, standardised and therefore suitable for use with different patients. Clinical implications of current injury thresholds are also discussed, in relation to obese patients and patients with muscle atrophy. PMID- 19678522 TI - UK perspectives on care in Russia. PMID- 19678523 TI - New post first impressions. PMID- 19678524 TI - Estimation of blood microcirculation in integuments by non-invasive speckle optical method under the photodynamic action. AB - In studies on animals (rats, rabbits) an experimental technique for estimation of microhemocirculation parameters of tissue under treatment has been developed. This technique has been used to compare the changes of speckle-optical parameters of skin microhemodynamics in laser-irradiated and light-isolated areas in the course of photodynamic therapy. Strong correlation between efficiency of tissue response and injection-photoirradiation time interval has been established. The results obtained are confirmed by the data of microhemodynamics estimation in the course of photodynamic therapy by the method of intravital microscopy. The defined ways of modification of the speckle-optical module will make it possible to optimize the conditions of parameters registration taking into account the object features and to improve informativity and sensitivity of the method. PMID- 19678525 TI - Blood flow quantification from 2D phase contrast MRI in renal arteries using an unsupervised data driven approach. AB - We present a clustering approach to segment the renal artery from 2D PC Cine MR images to measure arterial blood velocity and flow. Such information is important in grading renal artery stenosis and to support the decision on surgical interventions like percutaneous transluminal angioplasty. Results from 20 data sets (3 volunteers, 7 patients) show that the renal arteries could be extracted automatically and the corresponding velocity profiles were close (r = 0.977) to that obtained by manual delineations of the vessel areas. PMID- 19678526 TI - [Radon transfer and intracorporal deposition of radon decay products under balneotherapeutic conditions]. AB - The intracorporal deposition of radon decay products was determined on four persons after 40 and 30 min respectively in radon water with about 1500 Bq/L by wholebody gamma spectrometry. The measurements started about 2 1/2 h after exposure. In addition, the radon activity concentration of inspiratory and expiratory air was measured on one person during and after exposure and the deposition of radon decay products on the skin was measured on another person. The radon activity leaving the body with the expiratory air during exposure in the water (called "radon transfer") amounts to about 800 Bq. An intracorporal radon activity immediately after therapeutic exposure of about 3000 Bq was obtained as a result of first measurements by extrapolation from measurements starting about 2 1/2 hours later. Additional studies are necessary. There are indications that both the radon transfer and the intracorporal deposition can be increased by exposure in mixed radon-CO2 water. PMID- 19678527 TI - Experimental determination of peripheral photon dose components for different IMRT techniques and linear accelerators. AB - PURPOSE: To quantify the relative peripheral photon doses (PD) to healthy tissues outside the treated region for different IMRT technologies and linac head designs. MATERIAL AND METHODS: Measurements were performed on an Elekta linac for various energies (6 MV, 10 MV, 25 MV) at different depths at a distance of 29 cm off-axis (vertical measurements) and different distances from the field edge at constant depth of 10 cm (horizontal measurements). These measurements were compared with results obtained on a Siemens linac at 6 MV and 15 MV. TLD-700 detectors were used to quantify the PDs relative to the dose in the volume exposed with the primary beam. Intensity modulated (IM)-beams with identical fluence patterns were generated with a segmental multileaf (sMLC) technique and with lead-containing cerrobend compensators (MCP96). PD values of IM beams were compared with open beam values. All measurement results of the two different linacs, the different IM methods and the different energies were normalized to the same mean dose. RESULTS: PD values were distinctly higher near the surface (0.5-20 mm) than at larger depth and showed the same trend for all photon beam energies. In comparison with the open field, the photon dose component of PD for IM beams delivered with a segmental MLC technique were increased by a factor varying from 1.2 to 1.8, depending on photon energy and depth. This ratio was around 2 for compensator based IMRT. Depending on depth and distance from the field edge the PD on the Siemens machine was about 30% to 50% higher than on the Elekta machine for the same nominal photon energy. CONCLUSION: The treatment head design of a linac has a large impact on PD in IMRT as well as for open beams. PD can be minimized by proper selection of treatment delivery method and photon beam energy. PMID- 19678528 TI - Steep dose gradients for simultaneous integrated boost IMRT. AB - Steep dose gradients between two planning target volumes (PTVs) as may be required for simultaneous integrated boosts (SIB) should be an option provided by IMRT algorithms. The aim was to analyse the geometry of the SIB problem and to implement the results in an algorithm for IMRT segment generation denoted two step intensity modulated radiotherapy (2-Step IMRT). It was hypothesized that a gap between segments directed to the inner and the outer PTV would steepen the dose gradient. The mathematical relationships were derived from the individual dose levels and the geometry (diameters) of the PTVs. The results generated by means of 2-Step IMRT segments were equivalent or better than the segment generation using a commercial IMRT planning system. The dose to both the inner and the outer PTV was clearly more homogeneous and the composite objective value was the lowest. The segment numbers were lower or equal--with better sparing of the surrounding tissue. In summary, it was demonstrated that 2-Step IMRT was able to achieve steep dose gradients for SIB constellations. PMID- 19678529 TI - Dosimetry of a linear accelerator under respiratory gating. AB - In the present study, the impact of respiratory gating on the beam characteristics of a linear accelerator is investigated. The main focus is put on the influence of the duty cycle. Measurements were performed on a linear accelerator type Oncor (Siemens) with photon energies 6 MV and 15 MV, equipped with the Anzai gating system AZ-733V. Depth dose curves and beam profiles were found not to be significantly altered by gating even for duty cycles down to 5% for realistic respiration frequencies (dose variations all <2.5%). However, for very small duty cycles, the absolute dosimetry changes significantly (dose deviations > 10%), leading to clinically relevant underdoses. The crucial parameter for the dosimetry is the number of monitor units per gate. Our results imply that treatment planning for respiratory gating can be performed on the basis of data obtained under ungated operation if and only if the absolute gates sizes during treatment are sufficiently large. The limiting values for the gate sizes have to be determined individually for each accelerator. PMID- 19678530 TI - Alkaloids from Sternbergia colchiciflora. AB - Twenty-one alkaloids and related compounds were found in Sternbergia colchiciflora (Amaryllidaceae), a hitherto not studied plant species. Twenty of them were detected by GC-MS in the crude extracts of this plant species. Ten alkaloids were isolated and their structures confirmed by NMR, MS and CD measurements. Many of the compounds found in this species, such as lycorine, tazettine, haemanthidine, are known to possess strong bioactivity. Variations in the alkaloid pattern were found during the phenological cycle of the plant. Lycorine-type compounds were dominant in the plant organs during both the flowering period and dormancy. The alkaloid pattern during both periods of leaf development and fructification was dominated by haemanthamine-type in the leaves and lycorine-type compounds in the bulbs, respectively. PMID- 19678531 TI - Cytotoxicity of vincristine on the 5637 cell line is enhanced by combination with conferone. AB - Bladder cancer is one of the most common cancers worldwide, with the highest incidence in industrialized countries. There are three major histological subtypes of bladder cancer: transitional cell carcinoma (TCC) (> 90%), squamous cell carcinoma (< 10%) and adenocarcinoma (1-2%). The present study was carried out to assess the effects of conferone, a sesquiterpene coumarin isolated from Ferula badrakema, on a TCC subline, 5637 cells. In order to test the effects of conferone, 5637 cells were treated with different concentrations (16, 32, 64, 128 microg/ml) of conferone. The results indicated that conferone did not have any significant cytotoxic effect on these neoplastic cells. To determine the combining effects, the cells were cultured in the presence of different concentrations of conferone (16, 32, 64, 128 microg/ ml) and vincristine (30, 40, 50 microg/ml) in combination. The morphological changes were then observed and cytotoxicity effects were studied using the MTT assay 24, 48 and 72 h following drug administration. The cells were more rounded and granulated after treatments with both drugs in comparison to vincristine only. The results of the MTT assay confirmed the morphological observations. After 48 h of combined treatment with 40 microg/ml vincristine and 16 microg/ml conferone, the cytotoxicity of vincristine was increased by 23.6%. PMID- 19678532 TI - Anti-hepatitis B virus activity of new N4-beta-D-glycoside Pyrazolo [3,4 d]pyrimidine derivatives. AB - The reaction of 6-hydrazinyl-1,3-dimethylpyrimidine-2,4-(1H,3H)-dione (1) with ethoxymethylenemalononitrile afforded 5-amino-1-(1,3-dimethyl-2,6-dioxo-1,2,3,6 tetrahydropyrimidin-6-yl)-1H-pyrazole-4-carbonitrile (2). The latter was reacted with formamide and urea affording the corresponding 4-aminopyrazolo[3,4 d]pyrimidines 3 and 4. The reaction of monosaccharide aldoses with 3 and 4 gave stereoselectively the beta-N-glycosides 5a-d and 6a-d which were treated with acetic anhydride in pyridine to afford the corresponding acetylated derivatives 7a-d and 8a-d. The prepared compounds were tested for their antiviral activity against hepatitis B virus (HBV) and showed moderate to high activities. PMID- 19678533 TI - Hepatoprotective and antioxidant activity of two iridoids from Mussaenda 'dona aurora'. AB - Mussaenda 'dona aurora' (sepals) has been investigated for its hepatoprotective and antioxidant activities. The highest activity was observed in the ethyl acetate fraction. The separation of the ethyl acetate fraction gave two iridoids, sanshiside-D and lamalbide. Sanshiside-D exhibited a hepatoprotective activity greater than silimarin as was evidenced by significant reduction of ALT and AST in the serum enzyme levels. PMID- 19678535 TI - Antimicrobial activity of Gentiana lutea L. extracts. AB - Methanolic extracts of flowers and leaves of Gentiana lutea L., together with the isolated compounds mangiferin, isogentisin and gentiopicrin, were used to investigate the antimicrobial activity of the plant. A variety of Gram-positive and Gram-negative bacteria as well as the yeast Candida albicans has been included in this study. Both extracts and isolated compounds showed antimicrobial activity with MIC values ranging from 0.12-0.31 mg/ml. Our study indicated that the synergistic activity of the pure compounds may be responsible for the good antimicrobial effect of the extracts. Quantification of the secondary metabolites was performed using HPLC. PMID- 19678534 TI - Cassiaindoline, a new analgesic and anti-inflammatory alkaloid from Cassia alata. AB - Cassiaindoline is a new dimeric indole alkaloid isolated from Cassia alata L. leaves whose structure was elucidated through spectroscopic analyses. It exhibited analgesic activity at a dosage of 125.0 mg/kg mouse and decreased the number of writhings induced by acetic acid by 49.4%. It also showed a 57.1% anti inflammatory activity at a dosage of 75 mg/kg mouse. PMID- 19678536 TI - Xanthanolides with antitumour activity from Xanthium italicum. AB - Bioassay-guided fractionation of a CHCl3 extract of the leaves of Xanthium italicum Moretti led to the isolation of four xanthanolides: xanthatin (1), 4 epixanthanol (2), 4-epi-isoxanthanol (3), and 2-hydroxyxanthinosin (4). Their structures were determined by means of 1D and 2D NMR spectroscopy, including 1H 1H COSY, NOESY, HSQC and HMBC experiments, which resulted in complete and unambiguous 1H and 13C NMR chemical shift assignments. The isolated compounds 1-4 were evaluated for their antiproliferative activities, and were demonstrated to exert significant cell growth inhibitory activity against human cervix adenocarcinoma (HeLa), skin carcinoma (A431), and breast adenocarcinoma (MCF7) cells. PMID- 19678537 TI - A new tetrahydrofuran derivative from the endophytic fungus Chaetomium sp. isolated from Otanthus maritimus. AB - A hitherto unidentified endophytic strain of the genus Chaetomium, isolated from the medicinal plant Otanthus maritimus, yielded a new tetrahydrofuran derivative, aureonitolic acid (1), along with 5 known natural products, 2-6. The structure of 1 was determined by extensive spectroscopic analysis and comparison with reported data. Extracts of the fungus, grown either in liquid culture or on solid rice media, exhibited considerable cytotoxic activity when tested in vitro against L5178Y mouse lymphoma cells. Compounds 2 and 6 showed significant growth inhibition against L5178Y cells with EC50 values of 7.0 and 2.7 microg/mL, respectively, whereas 1 was inactive. PMID- 19678538 TI - Co-production of bisphenylpropanoid amides and meroterpenes by an endophytic Penicillium brasilianum found in the root bark of Melia azedarach. AB - A fungus, isolated from the root bark of Melia azedarach (Meliaceae), from which a series of meroterpenes have been reported, was identified as Penicillium brasilianum based on analysis of the ITS region of ribosomal DNA. From a rice culture of this fungus, the known phenylpropanoid amides brasiliamide A and B were obtained together with and a new, slightly modified congener, along with the meroterpenoids preaustinoid A1, preaustinoid B2 and austinolide. The compounds were isolated by the use of combined chromatographic procedures and identified by physical methods, mainly 1D and 2D NMR experiments, with distinction for 1H{15N} HMBC applied to brasiliamide A. The amides were tested for their antimicrobial activity and showed only weak inhibitory effects, against a set of pathogenic bacteria. PMID- 19678539 TI - Immunomodulatory and anti-inflammatory activity of selected osthole derivatives. AB - From osthole [7-methoxy-8-(3-methyl-but-2-enyl)-chromen-2-one] (I), obtained by selective extraction of Peucedanum ostruthium (L.) W. Koch roots, ostholic acid (II) was synthetized as a result of its oxidation with chromium trioxide. From ostholic acid, through its chloride, four amides were obtained: the morpholide 1, the p-chloro-benzylamide 2, the piperidine 3 and the N-methyl-piperazide 4. Except for 1, other compounds have not been described before. The amides 1-4 and their precursor osthole (I) were tested for their potential activities in selected immunological assays. The compounds showed moderate inhibitory activity in the humoral immune response to sheep erythrocytes in mice in vitro, and 4 was the most suppressive. The effects of 1 and 3 on concanavalin A- and pokeweed mitogen-induced mouse splenocyte proliferation were inhibitory and those of 4 stimulatory. The compounds were also tested for their activity on tumour necrosis factor a and interleukin 6 production, induced by lipopolysaccharide, in cultures of rat peritoneal cells and human peripheral blood mononuclear cells. Compounds 1, 3 and 4 inhibited tumour necrosis factor a (rat cells), whereas compound 2 stimulated the production of both cytokines. Compounds 1, 2 and 3 were also strongly inhibitory on tumour necrosis factor a production in human blood cells (73, 78 and 80% inhibition at 10 microg/ml, respectively). On the other hand, 2 and 4 stimulated the interleukin 6 production (2- to 3-fold stimulation). In addition, 2 and 4 suppressed the carrageenan-induced inflammation in mice (56.5% and 68.3% inhibition, respectively). In summary, the compounds predominantly displayed suppressive and antiinflammatory activities in the investigated models. PMID- 19678540 TI - The interleukin-18 inhibitory activities of echinocystic acid and its saponins from Impatiens pritzellii var. hupehensis. AB - Echinocystic acid (1), an echinocystic acid saponin, 2, and four of its ester saponins, 3-6, obtained from the active fraction of Impatiens pritzellii var. hupehensis, an traditional Chinese medicine for rheumatoid arthritis, were investigated for their effects on lipopolysaccharide (LPS)-induced interleukin (IL)-18 in human peripheral blood mononuclear cells. Three of them, 1, 2 and 6, showed obvious activity to inhibit the production of IL-18, especially the ester saponins with a sugar chain at C-28, 6. Structure-activity relationships are discussed in brief. PMID- 19678541 TI - Bioactive chemical constituents and comparative antimicrobial activity of callus culture and adult plant extracts from Alternanthera tenella. AB - Crude extracts of a callus culture (two culture media) and adult plants (two collections) from Alternanthera tenella Colla (Amaranthaceae) were evaluated for their antibacterial and antifungal activity, in order to investigate the maintenance of antimicrobial activity of the extracts obtained from plants in vivo and in vitro. The antibacterial and antifungal activity was determined against thirty strains of microorganisms including Gram-positive and Gram negative bacteria, yeasts and dermatophytes. Ethanolic and hexanic extracts of adult plants collected during the same period of the years 1997 and 2002 [Ribeirao Preto (SP), collections 1 and 2] and obtained from plant cell callus culture in two different hormonal media (AtT43 and AtT11) inhibited the growth of bacteria, yeasts and dermatophytes with inhibition halos between 6 and 20 mm. For the crude extracts of adult plants bioassay-guided fractionation, purification, and isolation were performed by chromatographic methods, and the structures of the isolated compounds were established by analysis of chemical and spectral evidences (UV, IR, NMR and ES-MS). Steroids, saponins and flavonoids (aglycones and C-glycosides) were isolated. The minimum inhibitory concentration (MIC) of the isolated compounds varied from 50 to 500 microg/mL. PMID- 19678542 TI - Selenium accumulation in mycelia of Flammulina velutipes during fermentation determined by RP-HPLC. AB - A method to estimate the content of selenium in organics was introduced based on reversed phase-high performance liquid chromatography (RP-HPLC).The maximum absorption peak of piazselenol was at 330 nm and the optimized temperature and pH value were 40 degrees C and 2.8, respectively. The minimum detection concentration of selenium(IV) was 0.06 microg/mL and the measurable range was 0.12-12.0 microg/mL. The organic selenium accumulation in golden needle mushroom (Flammulina velutipes) mycelia was obtained by subtracting the amount of inorganic selenium from that of total selenium. The organic selenium accumulation of various inoculation amounts showed that organic selenium accumulation in a unit volume of the fermentation broth was positively related the inoculation amount. Compared with the methods reported previously, the method used here is simple, reliable and less toxic. PMID- 19678543 TI - Ethylamine content and theanine biosynthesis in different organs of Camellia sinensis seedlings. AB - We examined the distribution of ethylamine, glutamic acid and alanine, which are utilized in theanine biosynthesis, and other major amino acids in leaves, stems, cotyledons and roots of 6-week-old tea seedlings. Ethylamine and glutamic acid, which are substrates of theanine synthetase, were distributed almost uniformly in all parts of the seedlings; the contents in micromol/g fresh wt varied from 0.44 0.88 (ethylamine) and 1.6-2.4 (glutamic acid). The content of alanine, a possible precursor of ethylamine synthesis, was significantly higher in roots (3.1 micromol/g fresh wt) than in other parts. Incorporation of radioactivity from [U 14C]-alanine into theanine was also higher in roots than in other organs. In 10 week-old seedlings, [1-14C]ethylamine was converted to theanine in young and developed leaves, stems, main and lateral roots; the highest rates of conversion were detected in the main and lateral roots. These results suggest that the theanine synthesis preferentially takes place in roots but is not restricted to them; substrates and the enzymatic machinery for theanine synthesis are available in all parts of tea seedlings. PMID- 19678544 TI - Cell viability and leakage of electrolytes in Avicennia germinans exposed to heavy metals. AB - The effect of heavy metal stress on the cell viability and leakage of electrolytes of Avicennia germinans leaf discs was investigated by the tissue tolerance test. Foliar discs were incubated with different Cd2+ or CU2+ concentrations for 24 h; thereafter, the cell membrane stability of the tissue was assayed by the cell viability Evans blue and leakage electrolytes methods. The results indicated that electrolyte leakage of the leaf discs increased 24 h after exposure to heavy metal stress, as shown by a reduction of the cell viability by 30% in discs exposed to higher doses of Cd2+ (0.546 M) and Cu2+ (0.7 M), respectively. Additionally, the histological analysis of the leaf discs exposed to heavy metal stress revealed that at higher Cd2+ and/or Cu2+ concentrations an increase in the intercellular spaces and destruction of mesophyll cells was observed 24 h after exposure. In summary, the biochemical and structural changes observed in foliar tissues of A. germinans suggest that higher cadmium and copper concentrations may result in structural changes and altered physiological characters in leaves. PMID- 19678545 TI - Neuroprotective and neurotrophic effects of isorosmanol. AB - The neurotoxicity induced by beta-amyloid (Abeta), which is one of the major causes of Alzheimer's disease (AD), leads to synaptic loss and subsequent neuronal death. Therefore, modulation of Abeta-induced neurotoxicity, as well as regeneration of damaged synapses could be important therapeutic approaches to control AD. In this study, we found that isorosmanol, an abietane-type diterpene, protected PC12 cells against Abeta-induced toxicity. Furthermore, isorosmanol promoted the generation of neurites. The neurotrophic effect of isorosmanol was enhanced by co-treatment with nerve growth factor (NGF). In addition, the neurite outgrowth induced by isorosmanol was accompanied by F-actin redistribution and increased expression of neurofilaments. Taken together, these results suggest that isorosmanol possesses both neuroprotective and neurotrophic effects, that might be beneficial for controlling AD. PMID- 19678546 TI - Some physicochemical peculiarities of poplar plastocyanins a and b. AB - The redox potentials of poplar plastocyanins a and b (PCa, PCb) were determined by spectrophotometric titrations of their reduced forms with [Fe(CN)6]3-. It was found that the two isoforms have the following millimolar extinction coefficients epsilon597 equilibrium constants Keq of one-electron exchange with [Fe(CN)6]4 /[Fe(CN)6]3-, and standard electron potentials E0: PCa: epsilon597 = (4.72 +/- 0.08) mM(-1) cm(-1), Keq = 0.133 +/- 0.009, E0' = (354 +/- 11) mV; PCb: epsilon597 = (5.23 +/- 0.16) mM(-1) cm(-1), Keq = 0.175 +/- 0.010, E0' = (363 +/- 12) mV. The pH dependence of the redox potential of PCb was studied too. It was found, that the value of E0' for PCb is constant in the pH range 6.5-9.5, but decreases in the range 4.8-6.5. On the whole, the dependence resembles that of PC from some well-known plant species, including poplar PCa. The changes of E0' in the pH-dependent region for poplar PCb, however, are smaller and are 13 mV per pH unit, whereas in the other well-known plant species the changes are about 50-60 mV per pH unit. It has been assumed that the weaker pH dependence of EO' of PCb accounts for some structural differences between PCa and PCb. PMID- 19678547 TI - Isolation and characterization of new Metschnikowia pulcherrima strains as producers of the antimicrobial pigment pulcherrimin. AB - Metschnikowia pulcherrima is a highly effective biocontrol yeast due to its pigment pulcherrimin that accumulates in the cells and in the growth medium. Three different strains of M. pulcherrima were isolated from local grapes. The yeast isolates were characterized on the basis of their biochemical, physiological and ITS1-5.8 s rDNA-ITS2 region. Based on the obtained results, the M. pulcherrima isolates were identified as new strains of M. pulcherrima. Strong antagonistic activities of the M. pulcherrima strains on the human pathogens Proteus vulgaris, Escherichia coli, Candida albicans, Candida parapsilosis, Candida krusei, and Trichosporon mucoides were determined. In addition, antagonistic effects of these M. pulcherrima strains were also tested against Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger, Trichoderma spp., Paecilomyces spp., and Bipolaris spp. and it was shown that the three different strains of M. pulcherrima also have an antagonistic effect on the growth of these fungal species at different extents. This study showed that all three strains of M. pulcherrima produce the same amount of the pigment pulcherrimin, but their antimicrobial activities on different microorganisms show important variations. PMID- 19678548 TI - Prevention of copper-induced calcium influx and cell death by prion-derived peptide in suspension-cultured tobacco cells. AB - Impact of copper on the oxidative and calcium signal transductions leading to cell death in plant cells and the effects of the copper-binding peptide derived from the human prion protein (PrP) as a novel plant-protecting agent were assessed using a cell suspension culture of transgenic tobacco (Nicotiana tabacum L., cell line BY-2) expressing the aequorin gene. Copper induces a series of biological and chemical reactions in plant cells including the oxidative burst reflecting the production of reactive oxygen species (ROS), such as hydroxyl radicals, and stimulation of calcium channel opening, allowing a transient increase in cytosolic calcium concentrations. The former was proven by the action of specific ROS scavengers blocking the calcium responses and the latter was proven by an increase in aequorin luminescence and its inhibition by specific channel blockers. Following these early events completed within 10 min, the development of copper-induced cell death was observed during additional 1 h in a dose-dependent manner. Addition of a synthetic peptide (KTNMKHMA) corresponding to the neurotoxic sequence in human PrP, prior to the addition of copper, effectively blocked both calcium influx and cell death induced by copper. Lastly, a possible mechanism of peptide action and future applications of this peptide in the protection of plant roots from metal toxicity or in favour of phytoremediation processes are discussed. PMID- 19678549 TI - Effect of DNA methylation on 18S rRNA gene sequences during culture of Taxus chinensis cells. AB - * Author for correspondence and reprint requests Z. Naturforsch. 64c, 418-420 (2009); received December 15, 2008 Cell suspension culture has rapidly become an alternative source of taxol, an anticancer compound. To investigate the role of DNA methylation in the cultural course of Taxus chinensis cells, analyses of 18S rRNA gene sequences of cultured T chinensis cells and related species were conducted. The phylogenetic analysis of 18S rRNA gene sequences indicated that HG 1 (the cultured T chinensis cells), like T mairei (the natural variety of T chinensis), should be a new variety of T chinensis, and cell culture can change the 18S rRNA gene sequence at the level of species despite 18S rRNA is the most conserved gene. The analyses of the CpG and TpG+CpA relative abundance and GC content of the 18S rRNA gene sequences made clear that DNA methylation contributed to changes of the 18S rRNA gene sequence of HG-1 at the level of species, which can make HG-1 to become a new variety of 7 chinensis. PMID- 19678550 TI - Micro-particle transporting system using galvanotactically stimulated apo symbiotic cells of Paramecium bursaria. AB - It is well known that Paramecium species including green paramecia (Paramecium bursaria) migrate towards the anode when exposed to an electric field in a medium. This type of a cellular movement is known as galvanotaxis. Our previous study revealed that an electric stimulus given to P bursaria is converted to a galvanotactic cellular movement by involvement of T-type calcium channel on the plasma membrane [Aonuma et al. (2007), Z. Naturforsch. 62c, 93-102]. This phenomenon has attracted the attention of bioengineers in the fields of biorobotics or micro-robotics in order to develop electrically controllable micromachineries. Here, we demonstrate the galvanotactic controls of the cellular migration of P bursaria in capillary tubes (diameter, 1-2 mm; length, 30-240 mm). Since the Paramecium cells take up particles of various sizes, we attempted to use the electrically stimulated cells of P bursaria as the vehicle for transportation of micro-particles in the capillary system. By using apo-symbiotic cells of P bursaria obtained after forced removal of symbiotic algae, the uptake of the particles could be maximized and visualized. Then, electrically controlled transportations of particle-filled apo-symbiotic P bursaria cells were manifested. The particles transported by electrically controlled cells (varying in size from nm to /m levels) included re-introduced green algae, fluorescence labeled polystyrene beads, magnetic microspheres, emerald green fluorescent protein (EmGFP)-labeled cells of E. coli, Indian ink, and crystals of zeolite (hydrated aluminosilicate minerals with a micro-porous structure) and some metal oxides. Since the above demonstrations were successful, we concluded that P bursaria has a potential to be employed as one of the micro-biorobotic devices used in BioMEMS (biological micro-electro-mechanical systems). PMID- 19678551 TI - Lipophilic compounds from the femoral gland secretions of male Hungarian green lizards, Lacerta viridis. AB - In spite of the importance of chemical signals (pheromones) in the reproductive behaviour of lizards, only a few studies have examined the role of specific chemical compounds as sexual signals. The secreted chemicals vary widely between species but whether this variation reflects phylogenetic or environmental differences remains unclear. Based on mass spectra, obtained by GC-MS, we found 40 lipophilic compounds in femoral gland secretions of male green lizards (Lacerta viridis), including several steroids, alpha-tocopherol, and esters of n C16 to n-C20 carboxylic acids, and minor components such as alcohols between C12 and C20, squalene, three lactones and one ketone. We compared these chemicals with those previously found in other closely related green lizard species, and discussed how phylogenetical differences and/or environmental conditions could be responsible for the differential presence of chemicals in different lizard species. PMID- 19678553 TI - Alkylation of adenosine deaminase and thioredoxin by acrylamide in human cell cultures. AB - Acrylamide is an alpha,beta-unsaturated vinyl monomer that causes cytotoxicity due to its alkylating properties. In recent years several proteins have been identified that are alkylated by acrylamide in vivo. This finding might explain the neurotoxic effects of acrylamide in humans. However, the list of potential acrylamide target proteins is far from being complete. In particular, the proteins that mediate the cytotoxicity of acrylamide in cell cultures remained unknown. Here we identify two novel acrylamide target proteins in human cell cultures (Jurkat, HepG2 and Caco-2), adenosine deaminase and thioredoxin. PMID- 19678552 TI - Effects of limonoids from Cipadessa fruticosa on fall armyworm. AB - Six mexicanolide limonoids isolated from the dichloromethane extract of the fruits of Cipadessa fruticosa Blume (Meliaceae) were evaluated against Spodoptera frugiperda (J. E. Smith). Gedunin was used as a positive control. When incorporated into an artificial diet of neonates at 50.0 mg kg(-1), febrifugin A showed 73.3% mortality. All the compounds showed moderate insecticidal activity, except for ruageanin A, when compared with the control. Febrifugin also showed growth inhibition and antifeedant activities (at 100.0 mg kg(-1)). The correlation between the insecticidal activity of the isolated compounds and their chemical structure was discussed. PMID- 19678554 TI - Inhibitors of ABC transporters and biophysical methods to study their activity. AB - Multidrug resistance caused by the presence and overproduction of ABC transporters makes serious problems in cancer treating. The drugs administered during therapy are pumped outside the cell using the energy obtained from ATP hydrolysis. The augmented dosage of drugs to overcome the multidrug resistance is not sufficient. Thus knowledge of the structure of ABC proteins is necessary to understand the rules of their action. It could be also helpful to understand how the multidrug resistance could be overcome. One of the strategies involves the treatment of cancer cells with a mixture of anticancer drugs and inhibitors of ABC transporters. The yeast Saccharomyces cerevisiae, whose PDR pumps are analogues of mammalian MDR proteins responsible for multidrug resistance, is a suitable research model. Biophysical methods with different fluorescent dyes seem to be very suitable for the measurement of the efflux pump activity. This review describes some known inhibitors of ABC proteins and biophysical methods which could be used for measuring the ABC transporters activity. PMID- 19678555 TI - A new mind-body approach for a total healing of fibromyalgia: a case report. AB - Fibromyalgia is a severe, chronic and widespread pain syndrome with no definite treatment protocol. Several medications are currently in use to treat this condition. Various pharmacological treatments, as well as alternative mind-body therapies, have been directed towards reducing fatigue and pain, but these treatments have only resulted in a partial relief of symptoms with no long-term or permanent effects. This study shows the results obtained from four female patients suffering from fibromyalgia after undergoing a mind-body treatment in which psychosocial genomic postulates as well as ideodynamic hand movements were the main tools employed in their healing. It is suggested that a mind-body oriented treatment could generate stable and permanent changes that enable patients to experience a total recovery from fibromyalgia. PMID- 19678556 TI - The effect of pregnancy on hypnotizability. AB - Hypnosis during pregnancy and childbirth has been shown to reduce labor analgesia use and other medical interventions. We aimed to investigate whether there was a difference in hypnotizability in pregnant and nonpregnant women. Study participants had hypnotizability measured by the Creative Imagination Scale (CIS) in the third trimester of pregnancy and subsequently between 14 and 28 months postpartum and when not pregnant. The 37 participants who completed the study gave birth in the largest maternity unit in South Australia between January 2006 and March 2007. CIS scores were increased in women when pregnant (Mean 23.5, SD 6.9) compared to when they were not pregnant (Mean 18.7, SD 6.6), p < 0.001. The mean effect size was 0.84 suggesting that the hypnotizability change was both statistically significant and clinically meaningful. Our study findings support previous evidence showing that women are more hypnotizable when pregnant than when not pregnant. PMID- 19678557 TI - A spiritual-hypnosis assisted treatment of children with PTSD after the 2002 Bali terrorist attack. AB - The aim of this study was to assess the effectiveness of a spiritual-hypnosis assisted therapy (SHAT) for treatment of posttraumatic stress disorder (PTSD) in children. All children, age 6-12 years (N=226; 52.7% females), who experienced the terrorist bomb blasts in Bali in 2002, and subsequently were diagnosed with PTSD were studied, through a longitudinal, quasi-experimental (pre-post test), single-blind, randomized control design. Of them, 48 received group SHAT (treatment group), and 178 did not receive any therapy (control group). Statistically significant results showed that SHAT produced a 77.1% improvement rate, at a two-year follow up, compared to 24% in the control group, while at the same time, the mean PTSD symptom score differences were significantly lower in the former group. We conclude that the method of spiritual-hypnosis is highly effective, economic, and easily implemented, and has a potential for therapy of PTSD in other cultures or other catastrophic life-threatening events. PMID- 19678558 TI - The hypnotic diagnostic interview for hysterical disorders, pediatric form. AB - This article reports on the use of hypnosis to facilitate the diagnostic process in two cases of pediatric hysterical reactions. The Hypnotic Diagnostic Interview for Hysterical Disorders (HDIHD), an interview tool, specifically designed for these cases, is reported. The first case was an adolescent male with motor Conversion Disorder manifested as paralysis of his lower limbs. The second was a preadolescent girl with sensory Conversion Disorder manifested as reduction of visual field in her right eye. Freudian conceptualization of hysterical reactions was employed as the conceptual basis in the formulation of these cases. This orientation posits hysterical phenomena a psychological defense employed by individuals exposed to traumatic experiences in order to effectuate a defense from intolerable affective material. The emotionally overwhelming material converts into physical reactivity free of the traumatic consequences by keeping the intolerable images and emotions deeply repressed within the subconscious. As the focus on these cases was diagnostic, treatment efforts were avoided. As it turned out, environmental interventions, based on the obtained information from the hypnotic interviews, extinguished the symptoms. The children were symptom free at follow-up. PMID- 19678559 TI - A comment on an alleged association between hypnosis and death: two remarkable cases. AB - Dr. Ewin recently reported his research on two "remarkable" cases where hypnosis performed by a lay hypnotist was allegedly associated with the death of the subject. Commentary is provided about both cases. In the first case, it seems clear that the death was co-incident to the hypnosis. In the second case, Dr. Ewin speculates that hypnosis may have been related to the subject's death following her experience in a stage hypnosis show. Instead, we propose that the alerting suggestion used to terminate the hypnosis (that "the subjects would feel 10,000 volts of electricity through the seat of their chairs"), not hypnosis per se, was inappropriate and may have specifically adversely affected this particular subject due to her phobia regarding electricity. Legal ramifications of these cases regarding the issue of informed consent are raised. It is concluded that these cases do not imply a duty to warn subjects/patients that one possible negative consequence of undergoing hypnosis is death. PMID- 19678560 TI - [Sequencing and analysis of the full coding sequence of Batai virus isolated in China]. AB - By RT-PCR and TAIL-PCR, the full coding region of Batai virus isolated in China (YN92-4 strain)was sequenced for the first time. According to the results, the genome of the virus contained three segments S, M and L of 947, 4,371 and 6,860 nucleotides, respectively. The S segment coded a nucleoprotein of 234 amino acids and a nonstructural protein of 102 amino acids, the M and L segments coded a precursor protein of 1 ,435 amino acids and RNA polymerase of 2,239 amino acids, respectively. Compared with the full coding sequence of Batai viruses isolated out of China, the S and M segments of YN92-4 and ON-7/B/01 showed the highest homology in nucleotide and amino acid sequenes with similarity of 97.7% (100%) and 95.7% (98%), respectively. Since there was no full coding sequence information on the L segment in GenBank for the reference, the L segment of YN92 4 was compared with that of Bunyamwera virus and the homology of nucleotide and amino acid was 73.5%and 81.6%, respectively. Phylogenetic analysis showed YN92-4 strain was clustered into one group with the prototype of Batai virus (MM2222). The results suggested that the YN92-4 strain had no occurrance of genetic reassortment (like Ngari virus) and was close to the Batai virus (ON-7/B/01 strain) isolated from cattle serum in Japan. PMID- 19678561 TI - [Genetic analysis of the complete env genes of HIV-1 from paid blood donors in Henan province]. AB - Complete HIV-1 env genes were amplified by nested PCR from uncultured peripheral blood mononuclear cells (PBMCs) DNA of 60 HIV-1 positive paid blood donors in Henan province, and the amplified full-length genes were sequenced. Twenty one full-length env genes were obtained, sequence analysis found that 15 of them had intact open reading frame (ORF). Fourteen sequences conformed to subtype B', their average genetic distance with the international reference sequence RL42 was 4.87% +/- 0.31%. One was subtype B, its genetic distance with the international reference sequence HXB2 was 5.43%. The amino acid sequences of these env genes were deduced according to their nucleotide sequences and extensive analysis and comparison of important structural motifs were performed. The results indicated that there was no drastic alteration in the number and position of potential N linked glycosylation sites among these 15 sequences. And the residues involved in forming the CD4 binding site were highly conserved. Genotype prediction of coreceptor usage based on V3 sequence and net charge suggested that most samples use CCR5 coreceptor. GPGR motif at the tetrapeptide crown in the V3 loop was most common in these samples and it was detected in 40% sequences. The cleavage site of gp120/gp41 was highly conserved, so Gp160 precursor of all isolates would be efficiently cleaved into the Gp120 and Gp41 subunits. The known neutralizing antibody binding sites for 2G12, IgG1b12, 4E10 and 2F5 were also highly conserved, it is expected that most of these isolates will be sensitive to neutralization by these antibodies. Further study to elucidate the correlation of the env genotype to functionally relevant motifs is necessary and that will aid vaccine and novel drug design. PMID- 19678562 TI - [Genetic variation of gag gene in HIV-1 subtype B infections from Henan and Shanxi provinces of China]. AB - The 109 whole blood samples were collected from HIV-1 infected former blood donors in Henan and Shanxi. The RNA templates were extracted from plasma and used for the full gag gene amplification and sequencing. The sequences were divided into 3 groups according to sampling year. The Entropy software was used to identify the amino acids with composition difference among different groups of amino acid sequences. The results showed that there existed 8 and 13 amino acid sites with the statistical significance difference, respectively, in sequences in year 2004 and 2005, compared to those in 2002. Among them, there existed 5 amino acid sites in two groups. Of 16 amino acid sites, the increasing polymorphism and the decreasing polymorphism along the sampling year were observed in 10 and 6 amino acid sites respectively. Of 10 sites with increased polymorphism, 8 sites were located in the CTL epitopes recognized and presented by the main HLA alleles existed in Chinese population. The 6 sites with decreasing polymorphism all existed in main domains of Gag proteins. PMID- 19678563 TI - [Effects of disulfide bridges in glycoprotein E1 on the membrane fusion activity of rubella virus]. AB - To reveal the effects of disulfide bridges in rubella virus glycoprotein E1 on the membrane fusion activity, the recombinant plasmid pBSK-SPE2E1 and site directed mutagenesis to mutate 11 cysteines individually in the ectodomain of E1 to remove a disulfide bridge from the wild-type E1 were constructed. All mutants and the wild-type plasmid were expressed on BHK-21 cell. Giemsa Staining was used to show the polykaryon formed in the transfected BHK-21 cells. The cell surface expression efficiency of the plasmids was assayed with fluorescence-activated cell sorter (FACS). Hemadsorption was performed to detect the receptor recognition activity of the recombinant plasmids. The results showed that all the 10 disulfide bridges in the ectodomain of E1 played an important role in the process of the membrane fusion. The removal of any disulfide bridge resulted in the loss of the fusion activity. The disulfide formed by the 5th and the 8th cysteine might be critical for the interaction of E1 and E2. While the disulfide bridges formed by the 3rd, the 4th, and the 13th might influence the membrane fusion activity of E1 directly. PMID- 19678565 TI - [Analysis of epidemiologic feature and genetic sequence of Sapovirus in China]. AB - To investigate epidemiologic feature and genetic variance of Sapovirus among children in China, fecal specimens were collected from children under 5 years old with acute diarrhea from Feb 2006 to Jan 2007 in nine provinces including Anhui, Fujian et al. A total of 1,110 fecal samples were detected for Sapovirus by reverse transcriptase-PCR (RT-PCR). Ten samples (0.9%) were positive for Sapovirus. The PCR products were then sequenced and analysed by phylogenetic tree. The results indicated that the detected Sapovirus strains were classified into two genogroups and three genotypes, including G I/1, G I/3, G II/3. PMID- 19678564 TI - [Construction of recombinant adenovirus co-expressing M1 and HA genes of influenza virus type A]. AB - Based on the human H5N1 influenza virus strain A/Anhui/1/2005, recombinant adenovirus co-expressing M1 and HA genes of H5N1 influenza virus was constructed using an internal ribosome entry site (IRES) sequence to link the two genes. The M1 and HA genes of H5N1 influenza virus were amplified by PCR and subcloned into pStar vector separately. Then the M1-IRES-HA fragment was amplified and subcloned into pShuttle-CMV vector, the shuttle plasmid was then linearized and transformed into BJ5183 bacteria which contained backbone vector pAd-Easy. The recombinant vector pAd-Easy was packaged in 293 cells to get recombinant adenovirus Ad-M1/HA. CPE was observed after 293 cells were transfected by Ad-M1/HA. The co-expression of M1 and HA genes was confirmed by Western-blot and IFA (immunofluorescence assay). The IRES containing recombinant adenovirus allowed functional co expression of M1 and HA genes and provided the foundation for developing new influenza vaccines with adenoviral vector. PMID- 19678566 TI - [Biological characteristics and sequence analysis of fusion genes of Newcastle disease virus isolates]. AB - Twenty Newcastle disease virus (NDV) strains were isolated from chickens and geese in the field outbreaks during 2005 and 2006 in some regions of Jiangsu and Guangxi province. Assessment of the virulence by MDT and ICPI, RT-PCR and sequence analysis of fusion protein gene were used to compare the properties of NDV isolates. The results indicated that MDT and ICPI of the isolates were 45.3h 58.2h and 1.61 - 2.00 respectively, which confirmed that the all NDV isolates were highly virulent. And their hemagglutinin were not resistant to heat and belonged to fast pattern of elution. The results of nucleotide sequencing and phylogentic analysis of fusion protein gene showed that the twenty strains shared homology from 79.7% to 100% among themselves, from 78.1% to 83.4% and from 80.2% to 90.1% with NDV LaSota, F48E8, respectively. The putative amino acid sequences of fusion protein at the cleavage sites of all the isolates were 112R-R-Q-R/K-R F117, with the motif characteristics of the virulent NDV strain, which was in accordant with the results of assessment of the pathogenicity. The phylogentic tree based on sequences of fusion protein gene variable regions (47-420nt) revealed that the 18 strains belonged to sub-genotype VIId and the others belonged to an old genotype III of NDV, revealing that subgenotype VIId virus was responsible for the NDV outbreaks in some regions of Jiangsu and Guangxi promince recently. PMID- 19678567 TI - [A specific PCR assay and a nested PCR assay for the screening of ovine pulmonary adenomatosis]. AB - Ovine pulmonary adenomatosis (OPA) is a naturally occurring contagious lung tumor of sheep which was caused by an exogenous retrovirus of sheep, jaagsiekte retrovirus (JSRV). Although no specific circulating antibodies against the virus coud be detected in infected sheep, exogenous JSRV proviral DNA sequences (exJSRV) and JSRV RNA transcripts could be detected in lung tumors, lymphoreticular system and peripheral blood mononuclear cells (PBMC) from sheep affected by OPA. The sheep genome carried 15 to 20 copies of endogenous retrovirus loci (enJSRV) that were similar to JSRV in structural genes but the divergene in U3. Therefore, primers specific for the U3 sequences of exJSRV were designed for the specific PCR and nested PCR (n-PCR). Sensitivity between specific PCR assay and n-PCR assay was compared by using serial dilutions of positive plasmid pJSRV-LTR in a background of 700ng sheep genome DNA. Sensitivity of n-PCR was ten-fold higher than specific PCR. The n-PCR was only available in blood test for detection of JSRV infected sheep and might be useful in epidemiological studies and disease control of OPA. PMID- 19678568 TI - [Phylogenetic analysis of the neuraminidase genes of subtype N1 viruses in domestic ducks in eastern China]. AB - To examine the phylogenetic information regarding the gene pool of AIV in domestic ducks in eastern China, the NA genes of twenty-six viruses isolated during 2002-2006, including two H1N1 strains, tenH3N1 strains and fourteen HSN1 strains, which reflected the predominant N1 subtype viruses were subjected to phylogenetic analysis. The results indicated that AIVs of N1 subtype circulating in domestic ducks in eastern China were undergoing a gradual evolution. Analysis of the deduced amino acid sequences revealed that NAs from all isolated H5N1 viruses had a 20-aa deletion in the stalk region (residues 49-68), whereas no deletion was seen in the NAs from other HA subtype viruses. The viruses of H3N1 and H1N1 might have a propensity for reassortment of NA genes, whereas no direct evidence of reassortment of NA gene was obtained in H5N1 viruses. PMID- 19678569 TI - [Establishment of RT- LAMP for rapid detection of foot-and-mouth disease virus]. AB - A rapid detection of foot-and-mouth disease virus (FMDV) was established by using reverse transcription loop-mediated isothermal amplification (RT-LAMP) method, meanwhile its specificity and sensitivity were assessed. The results showed that the FMDV RNA could be amplified by incubation at 65degrees C for only 1h using six primers designed based on FMDV polyprotein gene and the amplification products could be detected easily by naked-eye. There is no cross reaction with other virus such as SVDV, SFV and PPV by detecting their RNA samples. The detection limit of this method was found to be 10(-5) dilution of RNA sample which was 100-fold higher than that of PCR and 10-fold higher than real-time PCR. PMID- 19678570 TI - [Construction and identification of replicon vector derived from an infectious full-length cDNA clone of a Sindbis virus]. AB - To construct vector system of XJ-160 virus, a Sindbis virus isolated in China, recombinant vector pBRepXJ together with its helper plasmid pBR-H were derived from XJ-160 viral infectious clone pBR-XJ160 by overlap-PCR. To quantitatively and qualitatively verify the function of the replicon system, recombinant plasmids pSinRep-EGFP, pBRepXJ-EGFP, pSinRep-R and pBRepXJ-R were constructed by cloning report genes of enhanced green fluorescent protein (EGFP) or Renilla luciferase (R. luc) into pBRepXJ or pSinRep5, a commercial Sindbis vector. And in Vitro-synthesized RNA from expression vectors were electroporated into BHK-21 cells. The results indicated that the replicon vector system was capable of self replicating in host cell, and the expression efficiency of heterologous genes corresponded with that of the commercial Sindbis vector (pSinRep5). Our study laid the basis for developing alphavirus vector system with Chinese intellectual property. PMID- 19678571 TI - [Research advance of TRIM5alpha on structure and restriction mechanism of HIV-1 replication]. PMID- 19678572 TI - [Marek's disease virus encoded miRNAs--an update review]. PMID- 19678573 TI - [Adaptation of Oral Health Impact Profile (OHIP-14) index for measuring impact of oral health on quality of life in elderly to Serbian language]. AB - BACKGROUND/AIM: There is an evidence of increasing necessity to use indices for measuring impact of oral health on the elderly life quality for the purposes of the state-of-the art clinical dental practice. It is at least because data obtained in that way suggest whether a treatment is required, help in deciding upon a kind of dental treatment, as well as in evaluating the extent of success of the applied therapeutical procedures. The aim of the study was to translate into Serbian the Oral Health Impact Profile-14 (OHIP-14) index and to assess its validity in practice as its initial verification in the Serbian speaking areas. METHODS: The OHIP-14 index was translated into Serbian and check-up of its easy to-understand done in a group of 30 common elderly (above 65 years of age) with a total or partial prosthetic restoration. The study individuals did not fill in a questionnaire by themselves, instead the questiones were put orally. If needed, additional explanations were given with no any suggesting a possible answer. RESULTS: Considering the given answers and the shown need for additional explanations to some questions indicated that the question number 5 in the OHIP 14, namely "Have you been selfconscious because of your mouth or dentures?" (Serbian, "Da li ste bili samosvesni usled problema sa Vasim ustima i protezama?") cannot be adequately translated into Serbian because even 28 (93.3%) of the tested individuals were not able to understand it inspite the given explanations. Other questions were well understood, thus allowing to use the answers to them as score for a certain discomfort. Of all the tested, 13 (43%) denied any psychosocial disorders or experienced them only rarely, while 60% of the tested answered affirmately to the question "Do you consider your life generally less satisfactory due to the problems induced by teeth and dentures?" pointing out a significant impact of oral health condition on the quality of life. CONCLUSION: The OHIP-14 index renders valuable and dependable data on the quality of life of elderly with dentures, thus it is favorable to use it in practice. Due to difficulties in understanding the question number 5, however, it was ommited off the final Serbian language version. Anyhow, its validity in a more numerous test group should be evaluated, as well as other indices introduced into Serbian speaking area to let a comparison made and the most suitable questionnaire used. PMID- 19678574 TI - [Arthroscopic findings after shoulder dislocation]. AB - BACKGROUND/AIM: Recurrent instability of the shoulder joint is frequently difficult to differentiate from diseased or injured rotator cuff or tendon of the forearm flexor (m. biceps brachii). Shoulder joint arthroscopy has been only recently introduced into instable shoulder joint lesion examination. The aim of this study was to present and analyse an arthroscopic finding on instable shoulder joint in order to determine causes and mechanisms of instability, as well as principles of surgical treatment. METHODS: Arthroscopy of the shoulder joint was performed in 158 patients with at least one documented shoulder joint dislocation. These patients were divided into two groups. The group I included the patients with one to three dislocations, while the group II those with more than three dislocations. Preoperative diagnosis was based on anamnestic data and clinical examination using specific tests, and on the diagnosis of shoulder joint using radiography or computed tomography. RESULTS: Out of the total number of the patients 138 (87.34%) had injury of the anterior patellar brim, 119 (75.32%) had failure of the anterior capsule, 126 (79.75%) had compressive cartilage injury of the posterior part of the head of the upper arm bone (Hill-Sachs lesion), 102 (64.56%) had insufficiency of glenohumeral tendon, 11 (6.96) had complete cut of the rotator cuff, 23 (14.56%) had injury of the posterior patellar brim, 12 (7.59%) had injury of the upper anterior-posterior patellar brim (SLAP). CONCLUSION: According to the obtained results it could be concluded that there is no a unique injury that leads to shoulder joint instability. It is necessary to point out to the significance of anamnesis and clinical examination in making diagnosis. Arthroscopic diagnostics is indicated in clinically unreliable findings as an additional method for determining operative treatment. PMID- 19678575 TI - Antimicrobial susceptibility profiles of thermophilic campylobacters isolated from patients in the town of Nis. AB - BACKGROUND/AIM: In some clinical forms of human Campylobacter infections, such as prolonged diarrhea or associated with postinfections sequels, antibacterial treatment is necessary. The aim of the present study was to evaluate the antimicrobial susceptibility of thermophilic Campylobacter strains isolated from patients with diarrhea, as well as from patients with diarrhea followed by postinfections sequels, to drugs used in the therapy of enterocolitis, and to nalidixic acid used in laboratory identification and differentiation of thermophilic Campylobacter spp. METHODS: We studied the antimicrobial susceptibility profiles of 131 Campylobacter strains isolated from patients with diarrhea (122 strains), diarrhea associated with rheumatic disorders (8 strains), and one strain isolated from a patient with Guillain-Barre Syndrome following Campylobacter enterocolitis. Susceptibility testing to erythromycin, gentamicin, tetracycline, chloramphenicol, ciprofloxacin and nalidixic acid was performed by the agar dilution method. RESULTS: In the strains we investigated, resistance to gentamicin and chloramphenicol was not recorded, whereas a low rate of strains resistant to erythromycin (2.4%), a higher prevalence of strains resistant to tetracycline (9.9%), and a high level of resistance to ciprofloxacin (29.8%) and nalidixic acid (33.3%) were registered. All strains resistant to nalidixic acid were also resistant to ciprofloxacin. In addition, there was no difference in the occurrence of resistance between strains isolated from patients with diarrhea as compared to those isolated from patients with diarrhea followed by postinfection disorders. CONCLUSION: The fact that the most of Campylobacter strains were sensitive to erythromycin and all to gentamicin, makes erythromycin an antibiotic of choice in the treatment of Campylobacter diarrhea and gentamicin when parenteral therapy should be administered. Resistance to tetracycline and, especially, ciprofloxacin, necessitates antibiotic susceptibility testing. PMID- 19678576 TI - [Determination of infiltrative ductal breast carcinoma differentiation grade in biopsy imprints]. AB - BACKGROUND/AIM: In patients with breast carcinoma there are many risk factors for assessment of breast carcinoma maturity and prognosis. Besides histological type of differentiation, cytologic criteria for the evaluation grade of the differentiation of infiltrative ductal breast carcinomas are very important for prognosis. The aim of this study was to define cytologic criteria for grading of infiltrative ductal carcinomas of the breast. METHODS: . The imprints of intraoperative biopsies from 124 patients were studied. They were air-dried and stained by May-Grunwald Giemsa method. The features assessed were: the degree and type of cell clustering, nuclear diameter and pleomorphism, chromatin structure, number and features of nucleoli, the aspect of cytoplasm, noncellular background and the variability of cells and nuclei. According to these morphologic features the infiltrative ductal carcinomas of the breast could be classified into three grades of differentiation. RESULTS: Cytologic and histologic differentation grade revealed disagreement among 34.6% of the imprints. In 9 of total 23 histologicaly well differentiated carcinomas, cytological differentation grade was moderately differentiated. In 63 carcinomas with histologic differentiation grade II, cytologic differentation grade was good in 12 and poor in 16 carcinomas. CONCLUSION: Cytologic and histologic grading were not identical in 34.6% of the imprints what points out the need to further definition of diagnostic criteria, especially for grade II of differentiation. PMID- 19678577 TI - [Oesophageal-cancer-derived death in the population of Belgrade in a period 1989 2006]. AB - BACKGROUND/AIM: Oesophageal cancer is the sixth most common cause of death from all malignant tumors in the world (fifth in men, eighth in women). This cancer was estimated to account for about 529 000 new cases and about 442 000 deaths in the year 2007. In the year 2002 the highest standardized mortality rates (per 100 000 habitants) of oesophageal carcinoma were noticed in the East Asia (men/women: 18.8/7.7) and East Africa (18.6/7.8), while the lowest were noticed in the Middle Africa (1.4/0.2) and West Africa (1.3/0.5). The aim of this descriptive epidemiologic study was to analyze epidemiologic situation of oesophageal cancer in Belgrade population during the period 1989-2006, using mortality data. METHODS: Mortality data were collected from the City Organization for Statistics. In data analysis we used mortality rates which were standardized directly using those of the world population as the standard, and proportions. A denominator for mortality rates was calculated using the Belgrade population which was an average of the two latest register years (1991 and 2002). In order to analyze trend mortality from oesophageal cancer we used linear trend. RESULTS: In Belgrade deaths from oesophageal cancer accounted for about 5.2% of all malignant tumors of intestinal system in male population, and 2.4% in female population. This cancer is, according to standardized mortality rates (per 100 000 habitants), on the fifth place in Belgrade population behind colorectal, stomach, pancreatic, liver and cholecystic cancer. During the period 1989-2006 in Belgrade 44 persons died from oesophageal carcinoma on the average each year, mainly men (75%), and the rest were women (25%). In male population during the same period we noticed a significant increase in trend mortality (y = 1.61 + 0.06x, p = 0.001), while in female population the increase of mortality was not significant. The male/female oesophageal cancer mortality ratio was 3:1. Mortality rates for oesophageal cancer rise with age in both sexes and they are highest in the age group of 70 and more years. Significant increase in mortality from oesophageal cancer was noticed in age groups 20-29 and over 70 in male population, and age group 40-49 in female population. CONCLUSION: Increasing trend in oesophageal mortality suggests the necessity for improving measures of primary prevention including education about risk factors for this carcinoma (smoking, alcohol consumption, hot food and drinks), early diagnosis, and treatment. PMID- 19678578 TI - The value of sentinel lymphadenectomy in radical operative treatment of cervical cancer. AB - BACKGROUND/AIM: Therapy of the early stages of cervical carcinoma is surgical or radiation therapy, and for advanced stages chemoradiotherapy. Pelvic and paraaortic lymphadenectomy in early stages offers the most important prognostic factor for survival. To evaluate the method and possible influence on surgical and therapy of the disease, we performed sentinel node (SN) identification and excision during open radical hysterectomy and lymphadenectomy in stage Ib-IIa cervical carcinoma. METHODS: Fifty patients initially diagnosed with invasive squamous-cell cervical cancer stage Ib-IIa were included in the study. Only blue dye was used for sentinel node mapping. During the surgery sentinel nodes were identified and sent to histopathology separately from the other lymph nodes. After lymphadenectomy, radical hysterectomy was performed. RESULTS: The mean age of our fifty patients was 49.10 years (SD = 5.92), and the mean number of extracted lymph nodes per patient was 25.78 (SD = 5.58). The number of sentinel nodes identified per patient was between 0 and 5, mean 2.60 (SD = 1.54). There were no inframesenteric paraaortic sentinel nodes found among the patients. The dominant tumor grades were 1 and 2, 40% and 50% respectively, and 37 out of 50 patients (74%) had tumor diameter less than 2 cm. In four patients (8%) SN were not identified. In the rest of 46 patients the presence of SN was bilateral (19 patients, 38%) or unilateral (27 patients, 54%). Positive SN were found in 17 patients (34%), and negative in 29 patients (58%). Out of the whole group of patients (50), 21 of them (42%) had positive lymph nodes (LN). In the crosstab statistics, no differences were noticed in the group without SN found, in comparison with tumor grade and diameter. Finally, our test showed sensitivity of 85% (SE = 8%), specificity 100%, positive predictive value of 100%, negative predictive value of 89.6% (SE = 5.60%), and effectiveness of 93% (SE = 3.6%) regarding sentinel lymphadenectomy. CONCLUSION: This method of sentinel lymph node identification is simple, but not reliable enough to support further laparoscopic SN excision in order to make the final decision about the treatment of cervical cancer. PMID- 19678579 TI - Empirical antimicrobial therapy of acute dentoalveolar abscess. AB - BACKGROUND/AIM: The most common cause of acute dental infections are oral streptococci and anaerobe bacteria. Acute dentoalveolar infections are usually treated surgically in combination with antibiotics. Empirical therapy in such infections usually requires the use of penicillin-based antibiotics. The aim of this study was to investigate the clinical efficiency of amoxicillin and cefalexin in the empirical treatment of acute odontogenic abscess and to assess the antimicrobial susceptibility of the isolated bacteria in early phases of its development. METHODS: This study included 90 patients with acute odontogenic abscess who received surgical treatment (extraction of a teeth and/or abscess incision) and were divided into three groups: two surgical-antibiotic groups (amoxicillin, cefalexin) and the surgical group. In order to evaluate the effects of the applied therapy following clinical symptoms were monitored: inflammatory swelling, trismus, regional lymphadenitis and febrility. In all the patients before the beginning of antibiotic treatment suppuration was suched out of the abscess and antibiotic susceptibility of isolated bacteria was tested by using the disk diffusion method. RESULTS: The infection signs and symptoms lasted on the average 4.47 days, 4.67 days, and 6.17 days in the amoxicillin, cefalexin, and surgically only treated group, respectively. A total of 111 bacterial strains were isolated from 90 patients. Mostly, the bacteria were Gram-positive facultative anaerobs (81.1%). The most common bacteria isolated were Viridans streptococci (68/111). Antibiotic susceptibility of isolated bacteria to amoxicillin was 76.6% and cefalexin 89.2%. CONCLUSION: Empirical, peroral use of amoxicillin or cefalexin after surgical treatment in early phase of development of dentoalveolar abscess significantly reduced the time of clinical symptoms duration in the acute odontogenic infections in comparison to surgical treatment only. Bacterial strains isolated in early stages of dentoalveolar abscess showed high sensitivity to amoxicillin and cefalexin. PMID- 19678580 TI - [Oxidative status evaluation in elite karate athletes during training process]. AB - BACKGROUND/AIM: Oxidative stress is a state of disturbed balance between reactive oxygen species (ROS) and reactive nitrogen species (RNS) on the one hand, and antioxidative defense on the other. As a result of oxidative stress we have irreversible changes in cell function leading to pathological tissue changes, development of many diseases and fast aging process. Increased consumption of oxygen during exercise could be the cause of oxidative stress. The aim of the study was to establish oxidative status of elite karate athletes in the state of rest and after the loading, monitoring the parameters of oxidative stress and components of antioxidative defense in a training process. METHODS: During training process, a group of 30 elite karate athletes was included in the study of prevalence. They were males, from 16 to 30 years of age. The examination was conducted both in resting condition and after the loading. Oxidative status was determined by the level of superoxide anion radical (O2-), hydrogen peroxide (H2O2) as prooxidants, and enzymes of the antioxidant defanse system: superoxide dismutase (SOD) and catalase (CAT). RESULTS: There was a homogenous group of young athletes of normal weight and fats percentage. A significant decrease of O2 after the load was founded, while the levels of H2O2 was significantly increased. Activity of SOD was not significantly changed after exsercise while that of CAT was significantly increased. CONCLUSION: In the long run a programmed physical exercise does not lead to oxidative stress, but an excessive physical load may cause its occurrence. PMID- 19678581 TI - Cost-effectiveness of four immunomodulatory therapies for relapsing-remitting multiple sclerosis: a Markov model based on data a Balkan country in socioeconomic transition. AB - BACKGROUND/AIM: A cost-effectiveness analyses of immunomodulatory treatments for relapsing-remitting multiple sclerosis (RRMS) in developed countries have shown that any benefit from these drugs is achieved at very high cost. The aim of our study was to compare the cost-effectiveness of five treatment strategies in patients diagnosed with RRMS (symptom management alone and in combination with subcutaneous glatiramer acetate, intramuscular interferon beta-1a, subcutaneous interferon beta-1a, or intramuscular interferon [beta-1b) in a Balkan country in socio-economic transition. METHODS: The Markov model was developed based on the literature about effectiveness and on local Serbian cost calculations. The duration of a cycle in the model was set to a month. The baseline time horizon was 480 months (40 years). The societal perspective was used for costs and outcomes, and they were discounted for 3% annually. Monte Carlo micro simulation with 1000 virtual patients was done. RESULTS: Significant gain with immunomodulatory therapy was achieved only in relapse-free years, while the time spent in health states EDSS 0.0-5.5 was longer with symptomatic therapy only, and gains in life years and QALYs were only marginal. One QALY gained costs more than a billion of Serbian dinars (more than 20 million US dollars), making each of the four immunomodulatory therapies cost-ineffective. CONCLUSION: Our study suggests that immunomodulatory therapy of RRMS in a Balkan country in socioeconomic transition is not cost-effective, regardless of the type of the therapy. Moderate gain in relapse-free years does not translate to gain in QALYs, probably due to adverse effects of immunomodulatory therapy. PMID- 19678582 TI - [Comparative analysis of lower extremities tissue perfusion by the use of perfusion scintigraphy method after hyperbaric oxygenation and lumbar sympathectomy]. AB - BACKGROUND/AIM: Lumbar sympathectomy (LS) increases pheripheral blood flow primarily by the arteriolar vasodilatation within the skin vascular net. Increase in tissue nutrition takes place only in the distal blood vessels of the skin. Nevertheless, in some patients sympathectomy brings about improvement in ischemic ulcerations healing. Hyperbaric oxigenation (HBO) is a medical treatment in which a patient breathes 100% oxygen under pressure higher than atmospheric implemented in special units allowing the whole body be in a chamber. The aim of the study was to determine efficacy of the applied therapies for the treatment of inoperable occlussive lower extremities (LE) arteries diseases according to the obtained results. METHODS: The study included 30 patients divided into two grups (15 patients each) in which stenosis level of the lower extremities arteries had been determined using aortography due to further treatment with HBO and LS. All the patients were clinically examined, their objective condition evaluation based on claudication distance, pain in rest, skin and skin adnexa atrophy, and temperature and LE functionality, as well as exposed to perfusion scintigraphy prior to the treatment and within 30 days after the treatment finishing. RESULTS: Analysing patients' status prior to and after the treatments applied the number of patients with obvious improvement was higher in those treated by HBO than those treated by LS. Measuring claudication distance revealed significantly greater changes in patients treated by HBO (from 178.57 m to 754.76 m) than in those treated by LS (from 229 m to 253 m). Other clinical symptoms, such as parasthesia, status of the skin adnexes (hair, nails), skin colour and temperature were also improved after the treatment by HBO. CONCLUSION: The results obtained in this study confirm the advantages of HBO over LS in therapy of inoperable occlussive LE disease, so LS could be definitely abandoned as a choice for treating such disease. PMID- 19678583 TI - [Autologous stem cell transplantation in the treatment of Hodgkin's disease]. AB - BACKGROUND/AIM: High-dose chemotherapy with autologous stem cell transplantation (ASCT) has shown to produce long-term disease-free survival in patients with chemotherapy-sensitive Hodgkin disease. The aim of the study was to evaluate efficacy of ASCT in the treatment of Hodgkin's disease. METHODS: Between May 1997 and September 2008, 34 patients with Hodgkin's disease in median age of 25 (range 16-60) years, underwent ASCT. Autologous SCT were performed as consolidation therapy in one poor-risk patients with complete response (CR) and in 10 patients in partial remission (PR) after induction chemotherapy (32.5%), for chemosensitive relapse (CSR 1 and CSR 2) in 47% patients and in 20.5% patients with chemoresistant disease (CRD). All except one patient were in stage III/IV, extranodal site of disease had 24 patients and bulky disease had 10 patients. All the patients received a uniform preparatory regimen (BEAM). RESULTS: An overall response was achieved in 30 of 32 evaluated patients, with 62.5% in CR and 31.25% in PR. After applying radiotherapy, two patients with PR after ASCT reached CR. Median follow-up was 15.5 months (range 3-133 months). The probability of overall survival (OS) and progression-free survival (PFS) at a 3-year period for all patients was 51.9 % and 48.9%, respectively. For 22 patients in CR after ASCT, a 3-year DFS was 66.5%. Estimates of 2.5-year survival were 14.3%, 61.9% and 100% for CRD, CSR and for patients with CR/PR, respectively (p < 0.01). However, when patients undergoing consolidation were analyzed separately from those in CSR, no significant difference in OS and PFS was observed according to the disease status at ASCT. In univariate analysis for OS, PFS i DFS, extranodal site of disease and disease bulk had no predictive value. Twelve patients died. The main cause of death was Hodgkin's disease. Transplant-related mortality was 3.1%. One patient with CRD developed secondary acute myeloid leukemia and died 28 months after the transplantation. CONCLUSION: Autologous SCT is efficient as consolidation therapy in high-risk patients and in chemosensitive relapse, but it has no benefit in patients with chemoresistant disease. PMID- 19678584 TI - [Most common mistakes in measuring intraocular pressure]. PMID- 19678585 TI - Middle turbinate angiofibroma in an elderly woman. AB - BACKGROUND: Angiofibromas are histologically benign vascular tumors, originating from the nasopharynx, near by the area of sphenopalatine foramen. These neoplasms occur typically in male adolescents. Reports of primary extranasopharyngeal angiofibromas have appeared sporadically in the literature in English. We present the first case of an elderly woman with tumor arising from the middle turbinate, diagnosed as angiofibroma. CASE REPORT: A 63-year-old female presented with left sided nasal obstruction and epistaxis. Endoscopic evaluation revealed a polypoid mass arising from the anteroinferior portion of the left middle turbinate. Computed tomography (CT) scan showed a soft-tissue opacity that filled the anterior part of the left nasal cavity. After the endoscopic excision of the mass, postoperative pathohistological and immunohistochemical analysis confirmed the diagnosis of an angiofibroma. Two years later, the patient was free of symptoms and without endoscopic evidence of recurrence. CONCLUSION: Extranasopharyngeal angiofibromas arising from the nasal cavity are extremely rare tumors. Immunohistochemical analysis is very important in all doubtful cases, especially in those with atypical location. PMID- 19678586 TI - [Russian medical help during liberation and defence wars of Serbia in XIX and early XX century (1804-1917)]. PMID- 19678587 TI - Hand hygiene compliance: why hospitals aren't getting it right. PMID- 19678588 TI - Boosting hand washing with monitors, feedback. PMID- 19678589 TI - Consultant: scoring, PPR requirements still unclear. PMID- 19678590 TI - For ED bottlenecks, look at your OR scheduling. PMID- 19678591 TI - CMS hospital compare hits front page news. PMID- 19678592 TI - Effects of tiredness on visuo-spatial attention processes in elite karate athletes and non-athletes. AB - "Attentional" adaptations are fundamental effects for sport performance. We tested the hypothesis that tiredness and muscular fatigue poorly affect visuo spatial attentional processes in elite karate athletes. To this aim, 14 elite karate athletes and 11 non-athletes were involved in an isometric contraction exercise protocol up to muscular fatigue. Blood lactate and attention measurements were taken. Posner's test probed "endogenous" (i.e., internally planned allocation of spatial attention) and "reflexive" (i.e., brisk variation of endogenous spatial attention due to unexpected external stimuli) attention. Lactate and attentional measurements were performed before (Block 1, B1) and after the fatiguing exercise (B2) and at the end of a recovery period (B3). Compared to the non-athletes, the athletes showed a better performance in the fatigue protocol, confirmed by the higher absolute lactate values in B2. The correct responses in the "valid trials" probing "endogenous" attention were 92.4% (B1), 93.9% (B2), and 95.8% (B3) in the non-athletes, and 98.5%, 96.4%, 95.5% in the elite karate athletes. The correct responses in the "invalid trials" probing "reflexive" attention were 95.4%, 89.7%, 93.2% in the non-athletes, and 96.4%, 97.3%, 98.5% in the elite karate athletes. The percentage of correct responses in the "invalid" trials significantly decreased from B1 to B2 in the non-athletes but not in the elite karate athletes. In conclusion, tiredness and muscular fatigue do not affect "reflexive" attentional processes of elite karate athletes, which is crucial to contrast attacks coming from an unexpected spatial region. PMID- 19678593 TI - Does the default-mode functional connectivity of the brain correlate with working memory performances? AB - The "default-mode" network is an ensemble of cortical regions that are typically deactivated during demanding cognitive tasks in functional magnetic resonance imaging (fMRI) studies. Using functional connectivity analysis, this network can be studied as a "stand-alone" brain system whose functional role is supposed to consist in the dynamic control of intrinsic processing activities like attention focusing and task-unrelated thought generation and suppression. Independent component analysis (ICA) is the method of choice for generating a statistical image of the "default-mode" network (DMN) using a task- and seed-independent distributed model of fMRI functional connectivity without prior specification of node region extent and timing of neural activation. We used a standard graded working-memory task (n-back) to induce fMRI changes in the default-mode regions and ICA to evaluate to DMN functional connectivity in nineteen healthy volunteers. Based on the known spatial variability of the ICA-DMN maps with the task difficulty levels, we hypothesized the ICA-DMN may also correlate with the subject performances. We confirmed that the relative extent of the anterior and posterior midline spots within the DMN were oppositely (resp. positively in the anterior and negatively in the posterior cingulate cortex) correlated with the level of task difficulty and found out that the spatial distribution of DMN also correlates with the individual task performances. We conclude that the working memory function is related to a spatial re-configuration of the DMN functional connectivity, and that the relative involvement of the cingulate regions within the DMN might function as a novel predictor of the working-memory efficiency. PMID- 19678594 TI - Cyto-architecture and neuronal types of the dorsomedial cerebral cortex of the common Indian wall lizard, Hemidactylus flaviviridis. AB - The cyto-architecture and morphology of the neuronal types of the dorsomedial cortex of the lizard, Hemidactylus flaviviridis has been studied with the help of Cresyl violet staining and Golgi impregnation method. The dorsomedial cerebral cortex displayed three neuronal layers. Layer-I contains only few neuronal somas and also the dendrites ascending from the subjacent layers. Layer-II is characterized by two to three cell thick densely packed neuronal somas. Layer-III contains loosely packed neuronal somas and the dendrites and axon descending from layer-I and II. Below the layer-III an ependymal layer is observed just above the ventricle. Six classes of neurons were distinguished in the cellular layer of dorsomedial cortex of Hemidactylus flaviviridis: bitufted neurons, pyramidal neurons, inverted pyramidal neurons, bipyramidal neurons, multipolar neurons, and candelabra-like monotufted neurons. The pyramidal cells were large showing more or less single type present in the cellular layer. The multipolar neurons have mostly intracortical dendritic branching and connections. Bipyramidal neurons showed pyramidal appearance of their soma and send dendritic branches towards the superficial plexiform layer and deep plexiform layer. The candelabra-like monotufted neurons have very high dendritic branching. The comparison of the neuronal types of dorsomedial cortex of reptiles with the parahippocampal area of birds and CA3 region of mammalian hippocampus suggests possibility of their homology. PMID- 19678595 TI - The emotional control of action: ERP evidence. AB - It is known that unpleasant emotions can modulate the speed of involuntary movements, yet the effects of aversive stimulation on voluntary motor acts have not been systematically investigated. The effects of aversive stimulation on subsequent movement-related cortical activity were examined using a task invalving compatible and incompatible movements. Negative shifts in the timing of two motor event-related potentials were found subsequent to aversive stimulation for compatible movements only. With analogy to the Fear-Potentiated Startle Reflex, a reactive mechanism affecting covert action, these Fear-Potentiated Movement-Related Potentials (FPMPs) reflect preparation for overt actions performed to cope with particular types of danger. Further analysis revealed a positive deflection in the left frontal cortex prior the execution of incompatible movements, which may reflect inhibitory suppression of externally triggered imitative tendencies. PMID- 19678596 TI - Bispectral index correlates with regional cerebral blood flow during sleep in distinct cortical and subcortical structures in humans. AB - The relationship between the Bispectral Index (BIS), an EEG-based monitor of anesthesia, and brain activity is still unclear. This study aimed at investigating the relationship between changes in BIS values during natural sleep and regional cerebral blood flow (rCBF) variations, as measured by Positron Emission Tomography (PET). Data were obtained from six young, healthy, right handed, male volunteers (20-30 years old) using the H2(15)O infusion method. PET scans were performed both during waking and various stages of sleep. BIS values were monitored continuously and recorded during each PET scan. Positive correlations were detected between BIS and rCBF values in dorsolateral prefontal, parietal, anterior and posterior cingulate, precuneal, mesiofrontal, mesiotemporal and insular cortices. These areas belong to a frontoparietal network known to be related to awareness of self conscious sensory perception, attention and memory. BIS values also positively correlated with activity in brainstem and thalami, both structures known to be involved in arousal and wakefulness. These results show that BIS changes associated with physiological sleep depth co-vary with the activity of specific cortical and subcortical areas. The latter are known to modulate arousal, which in turn allows sustained thalamo cortical enhancement of activity in a specific frontoparietal network known to be related to the content of consciousness. Thus, although mainly derived from frontal EEG, BIS could represent a wider index of cerebral activity. PMID- 19678597 TI - To bundle or not to bundle: lawmakers explore the question. PMID- 19678598 TI - The implant positioning guide and the stent: part I. PMID- 19678599 TI - Case report: erythema ab igne in a patient with diabetic neuropathy. PMID- 19678600 TI - Dietary fat: quantity and quality. PMID- 19678601 TI - Alpha and beta thalassemia. AB - The thalassemias are a group of inherited hematologic disorders caused by defects in the synthesis of one or more of the hemoglobin chains. Alpha thalassemia is caused by reduced or absent synthesis of alpha globin chains, and beta thalassemia is caused by reduced or absent synthesis of beta globin chains. Imbalances of globin chains cause hemolysis and impair erythropoiesis. Silent carriers of alpha thalassemia and persons with alpha or beta thalassemia trait are asymptomatic and require no treatment. Alpha thalassemia intermedia, or hemoglobin H disease, causes hemolytic anemia. Alpha thalassemia major with hemoglobin Bart's usually results in fatal hydrops fetalis. Beta thalassemia major causes hemolytic anemia, poor growth, and skeletal abnormalities during infancy. Affected children will require regular lifelong blood transfusions. Beta thalassemia intermedia is less severe than beta thalassemia major and may require episodic blood transfusions. Transfusion-dependent patients will develop iron overload and require chelation therapy to remove the excess iron. Bone marrow transplants can be curative for some children with beta thalassemia major. Persons with thalassemia should be referred for preconception genetic counseling, and persons with alpha thalassemia trait should consider chorionic villus sampling to diagnose infants with hemoglobin Bart's, which increases the risk of toxemia and postpartum bleeding. Persons with the thalassemia trait have a normal life expectancy. Persons with beta thalassemia major often die from cardiac complications of iron overload by 30 years of age. PMID- 19678602 TI - Dietary fatty acids. AB - Fatty acids can be divided into four general categories: saturated, monounsaturated, polyunsaturated, and trans fats. Saturated fatty acids and trans fats are associated with an increased risk of coronary heart disease. Monounsaturated fatty acids and polyunsaturated fatty acids are associated with a decreased risk of coronary heart disease, although these associations are not uniformly supported in the literature. Omega-3 fatty acids, which are a type of polyunsaturated fatty acid, have been studied as potential therapy for a variety of medical conditions because of their suspected anti-inflammatory properties. Omega-3 fatty acids have also been shown to provide some benefit to patients with cystic fibrosis, and may have a protective effect against dementia. Physicians should counsel patients about the importance of avoiding hydrogenated oils and foods containing trans fats because of their association with coronary heart disease in observational studies. PMID- 19678603 TI - Diagnosing and treating hair loss. AB - Physicians should be careful not to underestimate the emotional impact of hair loss for some patients. Patients may present with focal patches of hair loss or more diffuse hair loss, which may include predominant hair thinning or increased hair shedding. Focal hair loss can be further broken down into scarring and nonscarring. Scarring alopecia is best evaluated by a dermatologist. The cause of focal hair loss may be diagnosed by the appearance of the patch and examination for fungal agents. A scalp biopsy may be necessary if the cause of hair loss is unclear. Alopecia areata presents with smooth hairless patches, which have a high spontaneous rate of resolution. Tinea capitis causes patches of alopecia that may be erythematous and scaly. Male and female pattern hair losses have recognizable patterns and can be treated with topical minoxidil, and also with finasteride in men. Sudden loss of hair is usually telogen effluvium, but can also be diffuse alopecia areata. In telogen effluvium, once the precipitating cause is removed, the hair will regrow. PMID- 19678604 TI - Hoarseness in adults. AB - Numerous conditions can cause hoarseness, ranging from simple inflammatory processes to more serious systemic, neurologic, or cancerous conditions involving the larynx. Evaluation of a patient with hoarseness includes a careful history, physical examination, and in many cases, laryngoscopy. Any patient with hoarseness lasting longer than two weeks in the absence of an apparent benign cause requires a thorough evaluation of the larynx by direct or indirect laryngoscopy. The management of hoarseness includes identification and treatment of any underlying conditions, vocal hygiene, voice therapy, and specific treatment of vocal cord lesions. Vocal hygiene education is an integral aspect of the treatment of hoarseness in most cases. Referral to a speech-language pathologist for voice therapy may be particularly helpful for patients whose occupation depends on singing or talking loudly or for prolonged periods. Voice therapy is an effective method for improving voice quality and vocal performance in patients with nonorganic dysphonia and for treating many benign pathologic vocal cord lesions. Referral for surgical or other targeted interventions is indicated when conservative management of vocal cord pathology is unsuccessful, when dysplasia or carcinoma is suspected, or when significant airway obstruction is present. PMID- 19678607 TI - FPIN's clinical inquiries. Hip pain in preschool-age children. PMID- 19678608 TI - Scalp rash in a newborn. PMID- 19678610 TI - Chemical mechanisms for skin sensitization by aromatic compounds with hydroxy and amino groups. AB - It is well-known that aromatic diamino-, dihydroxy-, and amino-hydroxy compounds, with NH(2) and OH groups in ortho- or para-positions relative to each other, are strong skin sensitizers. In this paper, we analyze published potency and cross reactivity data, whereby animals sensitized to one of these compounds are challenged with other compounds. The data are consistent with two parallel chemical reaction mechanisms: oxidation to electrophilic (protein reactive) quinones or quinone imines or formation of protein-reactive free radicals such as the Wurster salt, which can be formed by para-phenylene diamine. Compounds with NH(2) and OH groups meta to each other have also been found to be skin sensitizers, in some cases quite strong sensitizers. For these compounds, direct formation of quinones or quinone imines is not possible, and free radicals of the Wurster salt type are not favored. Here, we present a molecular mechanism to rationalize the sensitization potential of such compounds and, using the results of quantum mechanics calculations, show how this mechanism can explain observed structure-potency trends. PMID- 19678611 TI - Open sandwich-based immuno-transistor for label-free and noncompetitive detection of low molecular weight antigen. AB - In this study, we proposed a new detection method, open sandwich-based immuno field effect transistor (OS-FET) for label-free and noncompetitive detection of low molecular weight antigen. The principle of OS-FET is based on the detection of intrinsic molecular charges caused by the small antigen-dependent interchain interaction of separated V(L) and V(H) chains from a single antibody variable region using the field effect. Introducing V(H) chain and small antigen bisphenol A into the OS-FET with the immobilized V(L) chain on the gate, we could detect electrically and directly the binding of bisphenol A by V(H) and V(L) chains. Although the detection limit of OS-FET was 1 nM to detect bisphenol A for the standard deviation of control signal, the addition of isothiocyanobenzyl-EDTA with negative charges to the V(L) chain enhanced the detection limit to 1 pM. We could directly transduce the charge density changes based on the capture of target on the gate into the electrical signals using the OS-FET. The platform based on the FETs is suitable for a label-free, noncompetitive, and quantitative detection system for small antigen analysis in environmental, food, and clinical research. PMID- 19678612 TI - Cavity-enhanced stimulated raman scattering from short GaP nanowires. AB - We report the first observation of stimulated Raman scattering (SRS) from semiconductor nanowires (SNWs). Using continuous wave (CW) excitation (514.5 nm), very strong nonlinear SRS was observed in backscattering from short segments of crystalline GaP NWs with diameter d = 210 nm when the wire length L < 1.1 microm. The SRS intensity was found to increase dramatically with decreasing L down to 270 nm. The effective threshold pump power P(T) needed to observe the SRS is quite small. Indeed, we observe values of P(T) 3 orders of magnitude smaller than reported for bulk crystals of strongly nonlinear LiIO(3) and Ba(NO(3))(2) and 2 orders of magnitude smaller than that reported recently for SRS from "top-down" fabricated silicon microwaveguides. P(T) was observed to decrease linearly with decreasing L. Our data are discussed in terms of theoretical results developed for dielectric cavities. The quality factors Q for the short GaP cylindrical cavities studied here are estimated to be approximately 15,000. Our results suggest that SNWs may find applications as Raman lasers. PMID- 19678613 TI - Small molecule ligands for bulged RNA secondary structures. AB - A class of wedge-shaped small molecules has been designed, synthesized, and shown to bind bulged RNA secondary structures. These minimally cationic ligands exhibit good affinity and selectivity for certain RNA bulges as demonstrated in a fluorescent intercalator displacement assay. PMID- 19678614 TI - Highly diastereoselective three-component vinylogous Mannich reaction between isoquinolines, acyl/sulfonyl chlorides, and silyloxyfurans. AB - Reaction of an isoquinoline, a silyloxyfuran, and an acyl or sulfonyl chloride provides easy access to a wide variety of isoquinolinobutyrolactones with excellent yields and diastereoselectivites (R*,R* isomer), even in the case of formation of quaternary centers (i.e., R(3) or R(4) = Me). Moreover, the use of a chiral auxiliary allowed formation of a single stereoisomer in 96% yield. This represents the first examples of asymmetric vinylogous Mannich reactions on isoquinolinium salts. PMID- 19678615 TI - Viscoelastic properties of nanocrystalline films of semiconducting chalcogenides at liquid/liquid interface. AB - The interfacial shear rheological properties of a continuous single-crystalline film of CuS and a 3D particulate gel of CdS nanoparticles (3-5 nm in diameter) formed at toluene-water interfaces have been studied. The ultrathin films (approximately 50 nm in thickness) are formed in situ in the shear cell through a reaction at the toluene-water interface between a metal-organic compound in the organic layer and an appropriate reagent for sulfidation in the aqueous layer. Linear viscoelastic spectra of the nanofilms reveal solid-like rheological behavior with the storage modulus higher than the loss modulus over the range of angular frequencies probed. Large strain amplitude sweep measurements on the CdS nanofilms formed at different reactant concentrations suggest that they form a weakly flocculated gel. Under steady shear, the films exhibit a yield stress, followed by a steady shear thinning at high shear rates. The viscoelastic and flow behavior of these films that are in common with those of many 3D "soft" materials like gels, foams, and concentrated colloidal suspensions can be described by the "soft" glassy rheology model. PMID- 19678616 TI - Metalloporphyrin assemblies on pyridine-functionalized titanium dioxide. AB - Porphyrin adsorption on TiO2 nanoparticles has been achieved for multiple porphyrins, and in mixed porphyrin assemblies, via axial ligation to surface bound pyridine anchored by either para carboxylic or phosphonic functionalizations. Homogenous assemblies were prepared and characterized, while mixed metalloporphyrin assemblies were demonstrated by controlling the concentration ratios of respective porphyrins in the modifying solution. Evaluation of the assemblies using spectroscopic techniques and electrochemistry confirms high porphyrin retention, while exhibiting their surface bound optical and electrochemical properties. A thorough study is discussed where several metalloporphyrins have been evaluated (Ru(CO)OEP, Ru(CO)TPP, and ZnTPP) for relative comparisons and relationships to pyridyl axial binding strengths. The systematic study evaluates multiple background cases using either H2TPP, TiO2 modification with benzoic acid, or unmodified TiO2 to confirm the high affinity of Ru and Zn porphyrins for surface-anchored pyridyl sites. The simple method of step-by-step coordinative anchoring of porphyrins to TiO2 using small, commercially available molecules is highly adaptable for use in dye-sensitized solar cells (DSSC) where intimate contact between the absorbing dye and the semiconductor is required. DSSC devices with novel mixed porphyrin assemblies were shown to give higher power performance than DSSCs utilizing sensitization with only one type of porphyrin. PMID- 19678618 TI - One-pot synthesis of coumarin-based oxazabicyclic and oxazatricyclic compounds and their fluorescence redox switching properties. AB - An oxazabicycle and an oxazatricycle were efficiently synthesized by coupling of coumarins and N-alkylquinolium or isoquinolium salt to investigate their fluorescence redox-switching properties. Chemical reduction of the strongly fluorescent oxazabicycle and oxazatricycle results in the ring-opened products with a distinct decrease in emission intensity. Both resulting ring-opened species can be swiftly reverted to the original ring-closed forms by oxidation. PMID- 19678617 TI - Honeycomb-patterned films fabricated by self-organization of DNA-surfactant complexes. AB - In this paper, DNA-based honeycomb films were successfully constructed from a simple solution casting process at high relative humidity, through the encapsulation of DNA with a cationic surfactant ditetradecyldimethylammonium (DTDA). Cationic surfactants electrostatically attach to phosphate anions of DNA in a molar ratio of 1:1 in the DNA-DTDA complex. Investigation of the effects of substrate, concentration, and solvent on the morphology of the microporous films demonstrated the wide generality and high reproducibility of the formation of DNA DTDA self-organized microporous films. The morphology of the microporous structures can be adjusted by slightly changing the concentration of the complex solution. DNA exists in the double helical B-form in the microporous film confirmed by circular dichroism spectrum. Dye molecule rhodamine B was loaded into the DNA-DTDA complex, presenting a fluorescent microporous film. DNA-DTDA complex as a host material provides a new method for tailoring fluorescent microporous films. The DNA-based ordered honeycomb films should be attractive for use in optical and optoelectronic devices, separation, ion transportation, and biosensors. PMID- 19678619 TI - Modulation of the ESIPT emission of benzothiazole type dye incorporated in silica based hybrid materials. AB - A set of silica-based hybrid material samples was obtained, with different molar fractions of tetraethylorthosilicate and dimethyldimethoxysilane, as inorganic and organic sol-gel precursors, respectively. The hybrid material was used as a matrix to disperse the 2-(4'-amino-2'-hydroxyphenyl)benzothiazole, a fluorophore with large Stokes shift caused by an excited state intramolecular proton transfer phenomena (ESIPT). UV-vis and steady-state fluorescence in the solid state were applied in order to characterize the photophysical behavior of the dye. The set of samples was characterized by infrared spectroscopy, elemental analysis, and N(2) isotherms. The fluorescent pyrene dye was used as a probe for local polarity environment. For the first time, a modulation in the ESIPT fluorescence emission was achieved, being correlated to the methyl organic content of the hybrid matrix. The equilibrium between the conformers in the ground state is affected by changes in the matrix hydrophobicity. The replacement of silanol groups by methyl groups on matrix surface decreases the hydrogen bond capacity with the dye stabilizing the ESIPT conformer, in spite of the normal conformers, which will present a short wavelength emission band. PMID- 19678620 TI - On the formation of new reverse micelles: a comparative study of benzene/surfactants/ionic liquids systems using UV-visible absorption spectroscopy and dynamic light scattering. AB - The microenvironment of the polar core generated in different ionic liquid reverse micelle (IL RM) systems were investigated using the solvatochromic behavior of 1-methyl-8-oxyquinolinium betaine (QB) as an absorption probe and dynamic light scattering (DLS) technique. The novel RM systems consist of two different ILs--1-butyl-3-methylimidazolium tetrafluoroborate (bmimBF4) and 1 butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide (bmimTf2N)- sequestrated by two different surfactants--Triton X-100 (TX-100) and benzyl-n hexadecyldimethylammonium chloride (BHDC)--in order to make IL/surfactant/benzene RMs. The effect of the variation of Ws (Ws=[IL]/[surfactant]) on the QB spectroscopy was used to characterize these nonaqueous RMs. DLS results confirm the formation of these IL RM systems because increasing Ws increases the droplet sizes. Moreover it is demonstrated that the structure of the sequestrated ILs depends strongly on the type of surfactant use to create the RMs. PMID- 19678621 TI - Analysis of responsive characteristics of ionic-strength-sensitive hydrogel with consideration of effect of equilibrium constant by a chemo-electro-mechanical model. AB - A multiphysics model is presented in this paper for analysis of the influence of various equilibrium constants on the smart hydrogel responsive to the ionic strength of environmental solution, and termed the multieffect-coupling ionic strength stimulus (MECis) model. The model is characterized by a set of partial differential governing equations by consideration of the mass and momentum conservations of the system and coupled chemical, electrical, and mechanical multienergy domains. The Nernst-Planck equations are derived by the mass conservation of the ionic species in both the interstitial fluid of the hydrogel and the surrounding solution. The binding reaction between the fixed charge groups of the hydrogel and the mobile ions in the solution is described by the fixed charge equation, which is based on the Langmuir monolayer theory. As an important effect for the binding reaction, the equilibrium constant is incorporated into the fixed charge equation. The kinetics of the hydrogel swelling/deswelling is illustrated by the mechanical equation, based on the law of momentum conservation for the solid polymeric networks matrix within the hydrogel. The MECis model is examined by comparison of the numerical simulations and experiments from open literature. The analysis of the influence of different equilibrium constants on the responsive characteristics of the ionic-strength sensitive hydrogel is carried out with detailed discussion. PMID- 19678622 TI - Network of tris(porphyrinato)cerium(III) arranged on the herringbone structure of an Au(111) surface. AB - A network of bithiophene-modified tris(porphyrinato)cerium(III) triple-decker (TD) complexes was formed by pulse injection depositing on the Au(111) surface. The structure was observed with ultrahigh vacuum scanning tunneling microscopy (UHV-STM). High resolution UHV-STM images revealed the details of network structure of TD and the internal structure of both bithiophene and porphyrin moieties, indicating that the TD complexes undergo face-on adsorption. The alkyl chains on these bithiophenes not only make the TD molecules soluble in organic solvents, but also play a key role in preventing them from forming bonds between the thiophenes and the Au substrate, which enables the porphyrin molecular surface to be perpendicular to the substrate. These results suggest that the thiophene moiety can be used to control the adsorption and assembly of molecules on the herringbone structure of the Au(111) surface. PMID- 19678623 TI - Binary adsorption equilibrium of carbon dioxide and water vapor on activated alumina. AB - Adsorption equilibria of a CO2/H2O binary mixture on activated alumina F-200 were measured at several temperatures and over a wide range of concentrations from 4% to around 90% of the saturated water vapor pressure. In comparison with the single-component data, the loading of CO2 was not reduced in the presence of H2O, whereas at low relative humidity the adsorption of H2O was depressed. The binary system was described by a competitive/cooperative adsorption model where the readily adsorbed water layers acted as secondary sites for further CO2 adsorption via hydrogen bonding or hydration reaction. The combination of kinetic models, namely, a Langmuir isotherm for characterizing pure CO2 adsorption and a BET isotherm for H2O, was extended to derive a binary adsorption equilibrium model for the CO2/H2O mixture. Models based on the ideal adsorbed solution theory of Myers and Prausnitz failed to characterize the data over the whole composition range, and a large deviation of binary CO2/H2O equilibrium from ideal solution behavior was observed. The extended Langmuir-BET (LBET) isotherm, analogous to the extended Langmuir equation, drastically underestimated the CO2 loading. By incorporating the interactions between CO2 and H2O molecules on the adsorbent surface and taking into account the effect of nonideality, the realistic interactive LBET (R-LBET) model was found to be in very good agreement with the experimental data. The derived binary isosteric heat of adsorption showed that the heat was reduced by competitive adsorption but promoted by cooperative adsorption. PMID- 19678626 TI - Supramolecular synthons in noncovalent synthesis of a class of gelators derived from simple organic salts: instant gelation of organic fluids at room temperature via in situ synthesis of the gelators. AB - The supramolecular synthon approach has been employed to synthesize noncovalently a series of low molecular mass organic gelators (LMOGs) derived from benzylammonium salts of variously substituted benzoic acids. The majority of the salts (75%) prepared showed interesting gelation properties. Instant gelation of an organic fluid, namely methyl salicylate, was achieved at room temperature by using most of the gelator salts by in situ synthesis of the gelators. Table top rheology and scanning electron microscopy (SEM) were used to characterize the gels. Single crystal X-ray diffraction studies revealed the presence of both 1D and 2D supramolecular synthons. X-ray powder diffraction (XRPD) studies indicated the presence of various crystalline phases in the fibers of the xerogels. By using these data, a structure-property correlation has been attempted and the working hypothesis for designing the gelator has been reinforced. PMID- 19678624 TI - Utilization of lysozyme charge ladders to examine the effects of protein surface charge distribution on binding affinity in ion exchange systems. AB - A lysozyme library was employed to study the effects of protein surface modification on protein retention and to elucidate preferred protein binding orientations for cation exchange chromatography. Acetic anhydride was used as an acetylating agent to modify protein surface lysine residues. Partial acetylation of lysozyme resulted in the formation of a homologous set of modified proteins with varying charge densities and distribution. The resulting protein charge ladder was separated on a cation exchange column, and eluent fractions were subsequently analyzed using capillary zone electrophoresis and direct infusion electrospray ionization mass spectrometry. The ion exchange separation showed a significant degree of variation in the retention time of the different variants. Several fractions contained coelution of variants, some with differing net charge. In addition, several cases were observed where variants with more positive surface charge eluted from the column prior to variants with less positive charge. Enzymatic digest followed by mass spectrometry was performed to determine the sites of acetylation on the surface of the variants eluting in various fractions. Electrostatic potential maps of these variants were then generated to provide further insight into the elution order of the variants. PMID- 19678627 TI - Construction of oligonucleotide microarrays (biochips) via thioether linkage for the detection of bacterial meningitis. AB - Oligonucleotide-based arrays are increasingly becoming useful tools for the analysis of gene expression and single-nucleotide polymorphism. Here, we report a method that allows the direct immobilization of thiolated oligonucleotides onto an epoxy-activated glass surface via a stable thioether linkage under microwaves. The described chemistry efficiently immobilizes the probes via terminal thiol groups with uniform spot morphology. The thioether linkage could endure repeated PCR-like heat cycling with only 2.5% loss after 20 cycles, indicating that the chemistry can be used in integrated PCR/microarray devices. The highlighting feature of the proposed method is that the detection limit for the probe concentration can be reduced to 0.25 microM with 20-mer oligonucleotides. The efficiency of the projected method (approximately 33%) indicates its advantage over the existing standard methods, viz., NTMTA (approximately 9.8%), epoxide amine (approximately 9.8%) and disulfide (approximately 1.7%). The constructed microarrays were validated through the detection of base mismatches and bacterial meningitis. These features make the projected strategy ideal for manufacturing oligonucleotide arrays and detection of mismatches and bacterial diseases. PMID- 19678629 TI - Reporting analytical data. PMID- 19678631 TI - Multiscale modeling of coarse-grained macromolecular liquids. AB - A first-principle multiscale modeling approach is presented, which is derived from the solution of the Ornstein-Zernike equation for the coarse-grained representation of polymer liquids. The approach is analytical, and for this reason is transferable. It is here applied to determine the structure of several polymeric systems, which have different parameter values, such as molecular length, monomeric structure, local flexibility, and thermodynamic conditions. When the pair distribution function obtained from this procedure is compared with the results from a full atomistic simulation, it shows quantitative agreement. Moreover, the multiscale procedure accurately captures both large and local scale properties while remaining computationally advantageous. PMID- 19678628 TI - Phospholipid conjugate for intracellular delivery of peptide nucleic acids. AB - Peptide nucleic acids (PNAs) have a number of attractive features that have made them an ideal choice for antisense and antigene-based tools, probes, and drugs, but their poor membrane permeability has limited their application as therapeutic or diagnostic agents. Herein, we report a general method for the synthesis of phospholipid-PNAs (LP-PNAs) and compare the effect of noncleavable lipids and bioreductively cleavable lipids (L and LSS) and phospholipid (LP) on the splice correcting bioactivity of a PNA bearing the cell penetrating Arg9 group (PNA-R9). While the three constructs show similar and increasing bioactivity at 1-3 microM, the activity of LP-PNA-R9 continues to increase from 4-6 microM, while the activity of L-PNA-R9 remains constant and that of LSS-PNA-R9 decreases rapidly in parallel with their relative cytotoxicity. The activity of both LP-PNA-R9 and L PNA-R9 dramatically increased in the presence of chloroquine, as expected for an endocytotic entry mechanism. The constructs were also found to have CMC values of 1.0 and 4.5 microM, respectively, in 150 mM NaCl, pH 7 water, suggesting that micelle formation may play a hitherto unrecognized role in modulating toxicity and/or facilitating endocytosis. PMID- 19678630 TI - Exploring catalyst and solvent effects in the multicomponent synthesis of pyridine-3,5-dicarbonitriles. AB - The effects of an ionic base, tetrabutylammonium hydroxide (TBAH), and an amine base, piperidine, on the direct synthesis of pyridine-3,5-dicarbonitriles using a multicomponent reaction (MCR) from aldehydes, malononitrile, and thiols were systematically investigated. The amine base showed better results when the MCR was performed in ethanol, whereas employing the ionic base in acetonitrile resulted in similar yields but in a much shorter reaction time. A modified protocol to overcome the difficulty in the direct synthesis of pyridine-3,5 dicarbonitriles via the MCR from sterically hindered aldehydes using either base was realized by changing the reaction solvent from ethanol to acetonitrile. Mechanistically, the two catalysts were found to each promote different pathways in the final oxidation step of the penultimate product, 1,4-dihydropyridine 6. A reaction intermediate, Knoevenagel adduct 7, plays the major role in the amine base-catalyzed system, while in the presence of an ionic base, aerobic oxygen acts as the primary oxidant. PMID- 19678632 TI - Self-assembly behavior and doxorubicin-loading capacity of acylated carboxymethyl chitosans. AB - A new type of acylated carboxymethyl amphiphilic chitosan (ACC) with the use of acyl chain of varying lengths, from C(2) to C(12), and various degrees of acyl substitution was successfully synthesized and has been characterized in terms of its self-assembly behavior, structural stability, and drug encapsulation. The resulting nanostructure of the ACC nanoaggregates can be well manipulated through a control of hydrophobicity. Structural evolution of the self-assembled nanoaggregates is extensively characterized via (1)H NMR, FTIR, DSC, and TEM. A critical value of the hydrophobic effect, (X(DH) x X(Cn)), i.e., a product of "degree of acyl substitution" and "carbon number of acyl chain", can be employed as an indicator for structural variation of the nanoaggregates: when (X(DH) x X(Cn)) exceeded 1.5, the architecture of the nanoaggregates underwent a structural transformation from solid nanoparticle to hollow nanocapsules. The nanoaggregates exhibited an excellent colloidal and structural stability in aqueous medium. An improved affinity toward drug encapsulation, i.e., doxorubicin, can be technically designed according to the amphiphilic nature of the resulting nanoaggregates for drug delivery. PMID- 19678633 TI - Distance dependence of electron transfer kinetics for azurin protein adsorbed to monolayer protected nanoparticle film assemblies. AB - The distance dependence and kinetics of the heterogeneous electron transfer (ET) reaction for the redox protein azurin adsorbed to an electrode modified with a gold nanoparticle film are investigated using cyclic voltammetry. The nanoparticle films are comprised of nonaqueous nanoparticles, known as monolayer protected clusters (MPCs), which are covalently networked with dithiol linkers. The MPC film assembly serves as an alternative adsorption platform to the traditional alkanethiolate self-assembled monolayer (SAM) modified electrodes that are commonly employed to study the ET kinetics of immobilized redox proteins, a strategy known as protein monolayer electrochemistry. Voltammetric analysis of the ET kinetics for azurin adsorbed to SAMs of increasing chain length results in quasi-reversible voltammetry with significant peak splitting. We observed rate constants (k degrees (ET)) of 12-20 s(-1) for the protein at SAMs of shorter alkanethiolates that decays exponentially (beta = 0.9/CH(2) or 0.8/A) at SAMs of longer alkanethiolates (9-11 methylene units) or an estimated distance of 1.23 nm and is representative of classical electronic tunneling behavior over increasing distance. Azurin adsorbed to the MPC film platforms of increasing thickness results in reversible voltammetry with very little voltammetric peaks splitting and nearly negligible decay of the ET rate over significant distances up to 20 nm. The apparent lack of distance dependence for heterogeneous ET reactions at MPC film assemblies is attributed to a two-step mechanism involving extremely fast electronic hopping through the MPC film architecture. These results suggest that MPC platforms may be used in protein monolayer electrochemistry to create adsorption platforms of higher architecture that can accommodate greater than monolayer protein coverage and increase the Faradaic signal, a finding with significant implications for amperometric biosensor design and development. PMID- 19678634 TI - Diffusion Monte Carlo approaches for evaluating rotationally excited states of symmetric top molecules: application to H(3)O(+) and D(3)O(+). AB - An approach is described for evaluating energies and wave functions for rotationally excited states of symmetric top molecules using diffusion Monte Carlo methods. The approach is based on the fact that, for many systems, the rotation/vibration Hamiltonian can be modeled by terms that depend on the vibrational coordinates and powers of the components of the rotational angular momentum vector, J. In the case of symmetric top molecules with M = 0, the rotational part of the wave function is given by the tesseral harmonics. We construct rotationally excited states within the diffusion Monte Carlo approach by imposing nodal surfaces that are obtained from the roots of the tesseral harmonics. Results are presented for H(3)O(+) and D(3)O(+) with J class C > class A. It is likely that nanomolar inhibitors of all serine beta lactamases could be achieved by rational exploitation of diacyl phosphates. PMID- 19678666 TI - Intramolecular cooperativity in the reaction of diacyl phosphates with serine beta-lactamases. AB - Asymmetric diaroyl phosphates (ArCOOPO(2)(-)OCOAr', where Ar = Ph, Ar' = 4 biphenyl, 2-benzothiophenyl and 2-benzofuranyl) have been prepared, evaluated as serine (classes A, C, and D) beta-lactamase inhibitors, and compared with respect to the latter with their symmetric parents, where Ar = Ar'. The asymmetric compounds, in general, were found to react with the beta-lactamases in two modes, corresponding to different orientations with respect to the active site, whereby either of the two aroyl groups may acylate the enzyme to form two different inert acyl-enzymes, E-COAr and E-COAr' . In all cases, the asymmetric compounds, in one orientation, react more rapidly with the enzymes tested than one symmetrical parent but not both. From comparisons of activation free energy differences, it was found that the changes in free energy on changing from one aryl group to another, in either the acyl group or the leaving group, were not additive, i.e., that the effect of changing one aroyl group to another depended on the leaving group and vice versa. Thus, intramolecular cooperativity between the aroyl groups must exist, arising either from direct interaction between them or from protein mediated interaction or from a combination of both. Such cooperativity brings fresh opportunities and challenges to the search for novel beta-lactamase inhibitors. PMID- 19678667 TI - Salen-based [Zn2Ln3] complexes with fluorescence and single-molecule-magnet properties. AB - A family of four isostructural complexes with a V-shaped pentanuclear [Zn(2)Ln(3)] core of general formula [Zn(2)Ln(3)(m-salen)(3)(N(3))(5)(OH)(2)] [Ln(III) = Tb(III) (1), Eu(III) (2), Ho(III) (3), Dy(III) (4); m-salen = N,N' ethylenebis(3-methoxysalicylideneamine)] were isolated and structurally characterized. The fluorescence and magnetic measurements of the four compounds were investigated. Complex 1 exhibits strong fluorescence properties, while single-molecule-magnet behavior is seen in complex 4. PMID- 19678668 TI - Syntheses of the terpyridine-bipyridine linked binary ligands and structural and redox properties of their cobalt complexes. AB - New binary ligands consisting of 2,2':6',2''-terpyridine and 2,2'-bipyridine were synthesized through the linkage of various lengths of the polymethylene chain (1a 1d; n = 3-6, where n is the number of CH(2) units). Their Co(II) complexes, 2a-2d (Cl complexes) and 2a'-2d' (H(2)O complexes), were synthesized. They exclusively formed mononuclear complexes, which could be oxidized to Co(III) complexes in aqueous solutions after prolonged exposure to the air. The single crystals of 2b and Co(III) complex 2c'' were obtained, and their crystal structures were determined. Cyclic voltammograms of the Co(II) complexes showed the redox processes of the terpyridine, Co(II)/Co(I), and Co(III)/Co(II). The redox waves of Co(II)/Co(I) in 2a and 2a' were slightly different from those of other complexes because of the short polymethylene chain. PMID- 19678669 TI - Hierarchy of electronic properties of chemically derived and pristine graphene probed by microwave imaging. AB - Local electrical imaging using microwave impedance microscope is performed on graphene in different modalities, yielding a rich hierarchy of the local conductivity. The low-conductivity graphite oxide and its derivatives show significant electronic inhomogeneity. For the conductive chemical graphene, the residual defects lead to a systematic reduction of the microwave signals. In contrast, the signals on pristine graphene agree well with a lumped-element circuit model. The local impedance information can also be used to verify the electrical contact between overlapped graphene pieces. PMID- 19678671 TI - Cobalt nanoparticle-assisted engineering of multiwall carbon nanotubes. AB - New methods of processing multiwall carbon nanotubes (CNTs) are demonstrated in experiments in the transmission electron microscope (TEM). These include precisely controllable cutting, repairing, and interconnecting of different CNTs with the assistance of an encapsulated Co particle. All processes involve the interactions between the metal and graphitic shells that are driven by combined electrical biasing [using a scanning-tunneling microscope (STM)-TEM setup] of the CNT and focused electron-beam irradiation of a Co-containing region. In particular, we present two CNT soldering processes, that is, Co-joined and Co catalytic connections. The former process uses a Co particle as the central node to which two CNTs are covalently attached on the opposite sides, and the latter makes use of the segregation of new graphitic shells from the metal at the connecting site, resulting in CNT plumbing. We compare the mechanical robustness of both connection types by direct force measurements in the TEM using an integrated atomic force microscope (AFM) setup. They reveal a tensile strength of 4.2 and 31 GPa, respectively, thus demonstrating the superiority of the Co catalytic connection whose strength is already comparable to standard CNTs. In addition, all connected nanotubes show metallic conduction. The developed methods could be of particular importance in future nanoelectronic device technology. PMID- 19678672 TI - Inhibitory effect of Pb2+ on the transport cycle of the Na+,K+-ATPase. AB - The effect of Pb(2+) on the transport cycle of the Na(+),K(+)-ATPase was characterized in detail at a molecular level by combining electrical and biochemical measurements. Electrical measurements were performed by adsorbing purified membrane fragments containing Na(+),K(+)-ATPase on a solid-supported membrane. Upon adsorption, the Na(+),K(+)-ATPase was activated by carrying out concentration jumps of different activating substrates, for example, Na(+) and ATP. Charge movements following Na(+),K(+)-ATPase activation were measured in the presence of various Pb(2+) concentrations to investigate the effect of Pb(2+) on different ion translocating steps of the pump cycle. These charge measurements were then compared to biochemical measurements of ATPase activity in the presence of increasing Pb(2+) concentration. Our results indicate that Pb(2+) inhibits cycling of the enzyme, but it does not affect cytoplasmic Na(+) binding and release of Na(+) ions at the extracellular side at concentrations below 10 muM. To explain the inhibitory effect of Pb(2+) on the Na(+),K(+)-ATPase, we propose that Pb(2+) may interfere with the hydrolytic cleavage of the phosphorylated intermediate E(2)P, which occurs in the K(+)-related branch of the pump cycle. PMID- 19678673 TI - Zinc oxide nanocrystals stabilized by alkylammonium alkylcarbamates. AB - Nearly monodispersed, spherical ZnO nanocrystals were synthesized from the reaction of an amide precursor, [Zn(N(i)Bu(2))(2)](2), with hexylamine followed by reactions of the as-formed solution in a moist air flow. Extensive experiments were conducted to optimize the synthesis and to characterize the nanocrystals. The room temperature reactions led to 3.3-5.3 nm nanocrystals with the sizes increasing in direct proportion to the relative humidity. Purification afforded high yields of free-flowing nanocrystals that were dispersible in nonpolar solvents. The overall synthesis requires several days, but it results in multigram quantities of stable, redispersible nanocrystals. The nanocrystals were characterized using elemental analysis, X-ray diffraction (XRD), transmission electron microscopy (TEM), thermogravimetric analysis (TGA), X-ray photoelectron spectroscopy (XPS), solution and solid-state NMR, IR, UV-vis absorption, and photoluminescence spectroscopies. In addition to providing H(2)O to serve as the source of oxygen in the ZnO, the air flow adds CO(2) that converts the alkylamine into an alkylammonium alkylcarbamate, which serves as the surfactant. Elemental analysis, TGA, and XPS results established that the total number of N-hexyl fragments on a 3.7 nm nanocrystal was 200, where they exist as an equal number of anionic carbamates and cationic ammonium ions. The addition of pure hexylammonium hexylcarbamate to ZnO nanocrystals prepared by literature methods resulted in the formation of a product that was similar to the ZnO formed using [Zn(N(i)Bu(2))(2)](2). Larger nanocrystals up to 7.3 nm were also obtained by heating smaller nanocrystals in a mixture of hexylamine and toluene at 119 degrees C. PMID- 19678674 TI - Synthetic study on tetrapetalones: stereoselective cyclization of N-acyliminium ion to construct substituted 1-benzazepines. AB - The synthesis of the tetracyclic core of complex antibiotic tetrapetalones has been achieved in three steps starting from the simple intermediate gamma-hydroxy amide, which can be accessed through a high-yielding six-step sequence. The successful synthesis relies on a novel strategy based on the N-acyliminium ion cyclization. PMID- 19678675 TI - Fluorescence sensing of glucose using glucose oxidase modified by PVA-pyrene prepared via "click" chemistry. AB - Poly(vinyl alcohol)-pyrene-glucose oxidase (PVA-Py-GOx), a water-soluble polymer possessing both fluorescent and oxidant sites in the structure, is synthesized by "click" chemistry and modification processes and characterized. The morphology of PVA-Py-GOx was characterized with atomic force microscopy (AFM), and a heterogeneous morphology due to the incorporation of GOx was observed. The capability of PVA-Py-GOx to act as a bioprobe for fluorescence sensing of glucose is examined. The postulated fluorescence mechanism for glucose analysis is based on the consumption of glucose by dissolved oxygen and GOx present in the structure. Thus, the fluorescence intensity of pyrene groups of the probe increases by the elimination of fluorescence quenching by oxygen. Glucose concentration was analyzed quantitatively from 0.25 to 3.0 mM by the fluorescence measurement. The effect of pH and amount of PVA-Py-GOx was also studied. The proposed system was applied to analyze glucose in real samples and compared with those obtained from commercial kits. PMID- 19678676 TI - Synthesis of antitumor marine alkaloid discorhabdin A oxa analogues. AB - Discorhabdin A (1) exhibits a strong cytotoxic activity in vitro, but it is difficult to synthesize and handle due to the instability of its highly strained N,S-acetal structure. We then designed the analogues of discorhabdin A which also have strong cytotoxic activity and stability. The synthesis and examination of the biological activity of various types of stable discorhabdin A oxa analogues (2) were achieved. PMID- 19678677 TI - Plasmonics of 3-D nanoshell dimers using multipole expansion and finite element method. AB - The spatial and spectral responses of the plasmonic fields induced in the gap of 3-D nanoshell dimers of gold and silver are comprehensively investigated and compared via theory and simulation using the multipole expansion (ME) and the finite element method (FEM) in COMSOL, respectively. The E-field in the dimer gap was evaluated and compared as a function of shell thickness, interparticle distance, and size. The E-field increased with decreasing shell thickness, decreasing interparticle distance, and increasing size, with the error between the two methods ranging from 1 to 10%, depending on the specific combination of these three variables. This error increases several fold with increasing dimer size, as the quasi-static approximation breaks down. A consistent overestimation of the plasmon's fwhm and red shifting of the plasmon peak occurs with FEM, relative to ME, and it increases with decreasing shell thickness and interparticle distance. The size effect that arises from surface scattering of electrons is addressed and shown to be especially prominent for thin shells, for which significant damping, broadening, and shifting of the plasmon band is observed; the size effect also affects large nanoshell dimers, depending on their relative shell thickness, but to a lesser extent. This study demonstrates that COMSOL is a promising simulation environment to quantitatively investigate nanoscale electromagnetics for the modeling and designing of surface-enhanced Raman scattering (SERS) substrates. PMID- 19678678 TI - Highly enantioselective synthesis of chiral secondary amines by gold(I)/chiral Bronsted acid catalyzed tandem intermolecular hydroamination and transfer hydrogenation reactions. AB - A method for the synthesis of enantiomerically enriched secondary amines with excellent ee values through the tandem intermolecular hydroamination/transfer hydrogenation of alkynes using a "gold(I) complex-chiral Bronsted acid" protocol is developed. The catalysis works for a wide variety of aryl, alkenyl, and aliphatic alkynes as well as anilines with different electronic properties. PMID- 19678679 TI - Long-term improvements to photoluminescence and dispersion stability by flowing SDS-SWNT suspensions through microfluidic channels. AB - Shearing single-walled carbon nanotubes (SWNTs) coated with sodium dodecyl sulfate in microfluidic channels significantly increases the photoluminescence (PL) intensity and dispersion stability of SWNTs. The PL quantum yield (QY) of SWNTs improves by a factor of 3 for initially bright suspensions; on the other hand, SWNT QYs in a "poor" suspension improve by 2 orders of magnitude. In both cases, the QYs of the sheared suspensions are approximately 1%. The increases in PL intensity persist for months and are most prominent in larger diameter SWNTs. These improvements are attributed to surfactant reorganization rather than disaggregation of SWNTs bundles or shear-induced alignment. The results also highlight potential opportunities to eliminate discrepancies in the PL intensity of different suspensions and further improve the PL of SWNTs by tailoring the surfactant structure around SWNTs. PMID- 19678680 TI - Effect of storage conditions on carbon-centered radicals in soy protein products. AB - Using electron paramagnetic resonance (EPR) spectroscopy, the levels of carbon centered radicals in retail samples of isolated soy protein (ISP), soy protein concentrate (SPC), and powdered soy milk were estimated to contain from 6.12 x 10(14) to 1.98 x 10(15) spins/g of soy product. Roasted soy nuts contained about 5.70 x 10(15) spins/g. The peak to peak line width of the carbon-centered radicals from soy nuts was about 10 gauss, whereas ISP samples with a similar peak height had a peak to peak line width of about 8 gauss. Retail snack bars containing ISP, SPC, and/or roasted soy nuts with a total protein content of either 13, 21, or 29% contained 5.32 x 10(14), 6.67 x 10(14), and 5.74 x 10(14) spins/g of snack bar, respectively. Levels of carbon-centered radicals in newly prepared samples of ISP were much lower than levels in the retail soy protein products and levels previously reported for commercial ISP and laboratory ISP samples. The levels of radicals in ISP samples increased over a 12-25 week period of storage in the dark at 22 degrees C and exposed to air from about 8.00 x 10(13) spins/g immediately after preparation to 9.95 x 10(14) spins/g of ISP. Storing the ISP samples under nitrogen at 22 degrees C greatly reduced the increase in radical content, whereas storing the ISP in 99.9% oxygen at 40 degrees C accelerated the formation of stable carbon-centered radicals. ISP samples hydrated at either 22 or 92 degrees C, rapidly frozen, and dried lost about 92% of the trapped radicals. The level of carbon-centered radicals in these same ISP samples immediately began to increase during subsequent storage exposed to the air and gradually returned to similar levels obtained before they were hydrated. PMID- 19678681 TI - Catalytic enantioselective allylation of dienals through the intermediacy of unsaturated pi-allyl complexes. AB - The nickel-catalyzed enantioselective addition of allylboronic acid pinacol ester [allylB(pin)] to alpha,beta,gamma,delta-unsaturated aldehydes is described. This reaction results in a remarkable inversion of substrate olefin geometry, providing the Z,E-configured reaction product in good enantioselectivity and olefin stereoselectivity. The reaction appears to proceed by conversion of the dienal to an unsaturated pi-allyl complex followed by reductive elimination via transition state II. Enantioselectivities range from 73-94% ee for a range of delta-substituted dienals when chiral ligand L3 is employed. PMID- 19678682 TI - Removal of lipids, dioxins, and polychlorinated biphenyls during production of protein isolates from baltic herring ( Clupea harengus ) using pH-shift processes. AB - Dioxins and PCBs are toxic, lipophilic, and persistent substances that impose a serious health threat. A major risk of exposure to these toxic substances is consumption of fish from polluted waters, such as the Baltic Sea. The aim of this study was to investigate if pH-shift processing of Baltic herring with elevated toxicity levels could be used to produce a protein isolate with low fat content and, thereby, reduced dioxin and PCB levels. Both acid (pH 2.7) and alkaline (pH 11.2) pH-shift processing were investigated and resulted in efficient reduction of fat, dioxin, and PCB levels. A reduction of 70-80% per amount of protein was determined for all of these parameters. The amounts, and thus the removal, of lipids and dioxins (R(2) = 0.952) as well as lipids and PCBs (R(2) = 0.996) were highly correlated (p < 0.01). A mass balance of the alkaline pH-shift process showed that most of the fat and pollutants were found in the floating fat emulsion layer of the first centrifugation, followed by the pellet of the first centrifugation. These data show that the pH-shift protein isolation technique can be used to process herring with elevated dioxin and PCB levels and thereby increase the usage possibilities of such fish. PMID- 19678683 TI - A novel two-resistance model for description of the adsorption kinetics onto porous particles. AB - A novel two-resistance model for description of the sorption kinetics by porous particles has been proposed. The model takes into account two kinetic steps of different kinds which are involved in the overall sorption process rate: (i) the rate of solute diffusion in pores of the sorbent particles having uniform sizes and characterized by the homogeneous intraparticle diffusion coefficient and (ii) the rate of a direct adsorption/desorption process on the surface, described by applying the statistical rate theory (SRT) approach. Two different kinds of sorbent particles geometry are considered: the spherical and the plane particles, having their dimension characterized by radius and thickness, respectively. The meaning of the parameters which influence the sorption kinetics has been discussed. The results make it possible to judge which conditions have to be fulfilled to consider only one kinetic step and, thus, to simplify the theoretical description of a given system. The conclusion has been drawn that the concave character of kinetic sorption isotherms plotted in the function of the square root of time is of a general nature and is connected with the situation when at least two different processes are involved in controlling the sorption kinetics. PMID- 19678684 TI - Role of surfactants in the interaction of dye molecules in natural DNA polymers. AB - Solutions and powders formed from salmon sperm deoxyribonucleic acid (DNA) reacted with the cationic surfactant cetyltrimethylammonium chloride (CTMA-Cl) incorporated fluorescent rhodamine molecules: anionic sulforhodamine 640 (SRh) or cationic/zwitterionic rhodamine 640 perchlorate (RhP). The role of the cationic surfactant in the interaction between rhodamine dye and DNA-surfactant molecules has been investigated in both solution and solid state using optical spectroscopy and electrophoresis. Unexpectedly, the dye molecules did not interact directly with DNA, rather the DNA double helix acted as a template for the interaction between dye molecules and CTMA in the DNA/CTMA complex. The SRh and RhP molecules yield different fluorescence characteristics with increasing DNA/CTMA amount, indicating different configurations between the CTMA ligands. PMID- 19678685 TI - Phenylalanine substitution at site B10 (L29F) inhibits metmyoglobin formation and myoglobin-mediated lipid oxidation in washed fish muscle: mechanistic implications. AB - Wild-type sperm whale myoglobin (WT Mb) and L29F mutant were used to examine the effect of metMb formation rate on Mb-mediated lipid oxidation in washed cod muscle. MetMb formation was 15-fold slower in L29F compared to WT Mb at 2 degrees C (pH 5.7). The electrostatic interaction of bound O(2) and the partial positive edge of the phenyl ring of phenylalanine(29) inhibits deoxyMb-mediated metMb formation and may displace protons that promote oxyMb oxidation. Ferrous L29F was a poor promoter of lipid oxidation in washed cod during extended storage, whereas ferrous WT Mb oxidized the substrate readily. This diminishes a role for free radicals produced by the reaction of oxyMb with peroxides. MetL29F was an effective promoter of lipid oxidation. Mb mutants with low hemin affinity (H97A) were better promoters of microsomal lipid oxidation than mutants with higher hemin affinity (WT Mb and V68T). The much higher heme affinity of oxyMb compared to metMb partly explains the poor ability of ferrous L29F to oxidize lipids in washed cod at post-mortem pH during 2 degrees C storage. The relative roles of cross-linked Mb and hypervalent Mb species to promote lipid oxidation in washed cod at post-mortem pH are discussed. PMID- 19678686 TI - Preparation, characterization, and antiproliferative activities of the Se containing polysaccharide SeGLP-2B-1 from Se-enriched Ganoderma lucidum. AB - Inorganic selenite can be transformed into organic forms and bind to proteins and polysaccharides in Se-enriched submerged Ganoderma lucidum cultures. In the present study, a novel Se-containing polysaccharide, SeGLP-2B-1, was purified from the Se-enriched mycelia of G. lucidum and the antiproliferative activities against six human cancer cell lines were investigated. The Se content of SeGLP-2B 1 was 186.7 microg/g, which was 150-fold larger than that of the regular polysaccharide GLP-2B-1 (1.3 microg/g). SeGLP-2B-1 (1.06 x 10(6) Da) was composed of glucose, rhamnose, xylose, and galactose with a molar ratio of 1.000:0.652:0.443:0.227. SeGLP-2B-1 exhibited an approximately 10-fold stronger antiproliferative activity against six human cancer cell lines as compared to GLP 2B-1. Thus, Se is believed to play an important role in increasing the antiproliferative property of SeGLP-2B-1. These findings indicate that SeGLP-2B-1 may serve as a dietary Se supplement. PMID- 19678688 TI - Suzuki polycondensation with a hairpin monomer. AB - Two straight monomers were subjected to an AA/BB-type Suzuki polycondensation with a hairpin-shaped 1,8-anthrylene monomer as the counterpart leading to a novel polyarylene which should have the preferred conformation of a folded chain. The molar masses were determined by gel permeation chromatography and dynamic light scattering and found to be M(w) = 14,000 and M(n) = 7,000. MALDI-TOF MS analysis of a fraction provides a fingerprint of the step-growth nature of this polymerization. PMID- 19678687 TI - Strain-dependent photoluminescence behavior of CdSe/CdS nanocrystals with spherical, linear, and branched topologies. AB - The photoluminescence of CdSe/CdS core/shell quantum dots, nanorods, and tetrapods is investigated as a function of applied hydrostatic and non hydrostatic pressure. The optoelectronic properties of all three nanocrystal morphologies are affected by strain. Furthermore, it is demonstrated that the unique morphology of seeded tetrapods is highly sensitive to non-isotropic stress environments. Seeded tetrapods can thereby serve as an optical strain gauge, capable of measuring forces on the order of nanonewtons. We anticipate that a nanocrystal strain gauge with optical readout will be useful for applications including sensitive optomechanical devices and biological force investigations. PMID- 19678689 TI - Methodology for high accuracy contact angle measurement. AB - A new version of axisymmetric drop shape analysis (ADSA) called ADSA-NA (ADSA-no apex) was developed for measuring interfacial properties for drop configurations without an apex. ADSA-NA facilitates contact angle measurements on drops with a capillary protruding into the drop. Thus a much simpler experimental setup, not involving formation of a complete drop from below through a hole in the test surface, may be used. The contact angles of long-chained alkanes on a commercial fluoropolymer, Teflon AF 1600, were measured using the new method. A new numerical scheme was incorporated into the image processing to improve the location of the contact points of the liquid meniscus with the solid substrate to subpixel resolution. The images acquired in the experiments were also analyzed by a different drop shape technique called theoretical image fitting analysis axisymmetric interfaces (TIFA-AI). The results were compared with literature values obtained by means of the standard ADSA for sessile drops with the apex. Comparison of the results from ADSA-NA with those from TIFA-AI and ADSA reveals that, with different numerical strategies and experimental setups, contact angles can be measured with an accuracy of less than 0.2 degrees. Contact angles and surface tensions measured from drops with no apex, i.e., by means of ADSA-NA and TIFA-AI, were considerably less scattered than those from complete drops with apex. ADSA-NA was also used to explore sources of improvement in contact angle resolution. It was found that using an accurate value of surface tension as an input enhances the accuracy of contact angle measurements. PMID- 19678690 TI - A (13)C{(31)P} REDOR NMR investigation of the role of glutamic acid residues in statherin- hydroxyapatite recognition. AB - The side chain carboxyl groups of acidic proteins found in the extra-cellular matrix (ECM) of mineralized tissues play a key role in promoting or inhibiting the growth of minerals such as hydroxyapatite (HAP), the principal mineral component of bone and teeth. Among the acidic proteins found in the saliva is statherin, a 43-residue tyrosine-rich peptide that is a potent lubricant in the salivary pellicle and an inhibitor of both HAP crystal nucleation and growth. Three acidic amino acids-D1, E4, and E5-are located in the N-terminal 15 amino acid segment, with a fourth amino acid, E26, located outside the N-terminus. We have utilized (13)C{(31)P} REDOR NMR to analyze the role played by acidic amino acids in the binding mechanism of statherin to the HAP surface by measuring the distance between the delta-carboxyl (13)C spins of the three glutamic acid side chains of statherin (residues E4, E5, E26) and (31)P spins of the phosphate groups at the HAP surface. (13)C{(31)P} REDOR studies of glutamic-5-(13)C acid incorporated at positions E4 and E26 indicate a (13)C-(31)P distance of more than 6.5 A between the side chain carboxyl (13)C spin of E4 and the closest (31)P in the HAP surface. In contrast, the carboxyl (13)C spin at E5 has a much shorter (13)C-(31)P internuclear distance of 4.25 +/- 0.09 A, indicating that the carboxyl group of this side chain interacts directly with the surface. (13)C T(1rho) and slow-spinning MAS studies indicate that the motions of the side chains of E4 and E5 are more restricted than that of E26. Together, these results provide further insight into the molecular interactions of statherin with HAP surfaces. PMID- 19678691 TI - Molecular structures of citrate and tricarballylate adsorbed on alpha-FeOOH particles in aqueous suspensions. AB - In this work, the adsorption of citric (2-hydroxypropane-1,2,3-tricarboxylic acid) and tricarballylic (propane-1,2,3-tricarboxylic acid) acids onto alpha FeOOH (goethite) in aqueous suspensions was studied as a function of pH and total ligand concentration in 0.1 M NaCl at 25.0 degrees C, and the molecular structures of the surface complexes formed were analyzed by means of ATR-FTIR spectroscopy. The adsorption experiments were carried out as a series of batch experiments, and a newly developed simultaneous infrared and potentiometric titration technique was used to collect in situ infrared spectra with high signal to-noise ratios. The high quality of the infrared spectra allowed analysis by means of two-dimensional correlation spectroscopy formalism that aided the resolution of pH-dependent spectral features. This has enabled the detection of two previously unidentified citrate-goethite surface complexes: one protonated species at low pH, and one inner sphere complex prevailing at high pH and coordinated via a combination of hydroxyl and carboxylate groups. In addition, an inner sphere complex involving only carboxylate coordination predominating at low pH and an outer sphere complex existing in the circumneutral pH region were identified. The behavior of tricarballylate parallels that of citrate, except no inner sphere surface complex is formed at high pH values, which is in accordance with the lack of an alpha-hydroxyl group. The comparison between citrate and tricarballylate reinforces previous observations showing that inner sphere surface complexes of pure carboxylates at water-iron oxide interfaces are suppressed at high pH values, where outer sphere species are relatively more predominant. It also shows that significant amounts of inner sphere surface complexes of carboxylates only seem to form in the basic pH region when the ligands contain complementary functional groups, such as the hydroxyl or amine groups. PMID- 19678692 TI - Excited states of DNA base pairs using long-range corrected time-dependent density functional theory. AB - In this work, we present a study of the excitation energies of adenine, cytosine, guanine, thymine, and the adenine-thymine (AT) and guanine-cytosine (GC) base pairs using long-range corrected (LC) density functional theory. We compare three recent LC functionals, BNL, CAM-B3LYP, and LC-PBE0, with B3LYP and coupled cluster results from the literature. We find that the best overall performance is for the BNL functional based on LDA. However, in order to achieve this good agreement, a smaller attenuation parameter is needed, which leads to nonoptimum performance for ground-state properties. B3LYP, on the other hand, severely underestimates the charge-transfer (CT) transitions in the base pairs. Surprisingly, we also find that the CAM-B3LYP functional also underestimates the CT excitation energy for the GC base pair but correctly describes the AT base pair. This illustrates the importance of retaining the full long-range exact exchange even at distances as short as that of the DNA base pairs. The worst overall performance is obtained with the LC-PBE0 functional, which overestimates the excitations for the individual bases as well as the base pairs. It is therefore crucial to strike a good balance between the amount of local and long range exact exchange. Thus, this work highlights the difficulties in obtained LC functionals, which provides a good description of both ground- and excited-state properties. PMID- 19678693 TI - Catalytic mechanism of DNA backbone cleavage by the restriction enzyme EcoRV: a quantum mechanical/molecular mechanical analysis. AB - Endonucleases, such as the restriction enzyme EcoRV, cleave the DNA backbone at a specific recognition sequence. We have investigated the catalytic mechanism of backbone phosphodiester hydrolysis by the restriction enzyme EcoRV by means of hybrid quantum mechanical/molecular mechanical calculations. An exhaustive computation of different reaction pathways is performed, thus generating a network of pathways. Comparison of the computed (AM1d/MM) enzymatic reaction pathways with an analogous mechanism for small-molecule model systems [AM1/d and B3LYP/6-31++G(d,p)] reveals that the transition barriers for associative hydrolysis, which is more probable in the model systems, are not lowered by the enzyme. Instead, a reaction mechanism which has mostly dissociative character is more likely. The protein environment is tuned to significantly electrostatically stabilize the transition state structures. The direct catalytic impact of essential residues is determined: The magnesium metal ion activates a water molecule, thus facilitating protonation of the leaving group. A reduction of the coordination number of the magnesium metal ion from six to four upon the positioning of the attacking water molecule explains why larger metal ions, such as calcium, are not catalytically active. The nucleophile is generated by the transfer of a proton from the attacking water molecule to a carboxylic oxygen atom of aspartate 90. The catalytic effect of lysine 92 involves proper positioning of the scissile phosphate group and, more importantly, stabilization of the metaphosphate intermediate in an orientation optimal for attack of the nucleophile. PMID- 19678694 TI - CH/pi interaction in benzene and substituted derivatives with halomethane: a combined density functional and dispersion-corrected density functional study. AB - The present work intends to establish the efficiency of dispersion-corrected density functionals in explaining the potential energy curves of benzene-methane, benzene-fluoroform, and 1,3,5-trifluoro benzene-methane complexes. The interaction energies of all of the complexes under investigation have been evaluated using both van der Waals-corrected and normal gradient-corrected Perdew Burke-Ernzerhof and Becke-Lee-Yang-Parr density functionals. Our analyses suggest that the potential energy curves for both benzene-methane and benzene-fluoroform complexes are in excellent agreement with highly accurate coupled cluster (CCSD(T)) results as well as high-level counterpoise-corrected MP2 results. Remarkably, the interaction energies of the benzene-fluoroform complex are fairly higher than those of the other two complexes. This is primarily attributed to the dispersion correction present in it. Finally, the overall study highlights the importance of halogen substitution in strengthening the CH/pi interactions. PMID- 19678695 TI - Orotic acid decarboxylation in water and nonpolar solvents: a potential role for desolvation in the action of OMP decarboxylase. AB - OMP decarboxylase (ODCase) generates a very large rate enhancement without the assistance of metals or other cofactors. The uncatalyzed decarboxylation of 1 methylorotate in water is shown to involve the monoanion, although uncharged 1 methylorotic acid is decarboxylated at a similar rate. To measure the extent to which the rate of the nonenzymatic decarboxylation of orotate derivatives might be enhanced by their removal from solvent water, the 1-phosphoribosyl moiety of OMP was replaced with 1-substituents that would allow it to enter less polar solvents. When the tetrabutylammonium salt of 1-cyclohexylorotate was transferred from water to a series of dipolar aprotic solvents, its rate of decarboxylation increased markedly, varying with the relative ability of each solvent to release the substrate in the ground state from stabilization by solvent water acting as a proton donor. These findings are consistent with the view that separation of the substrate from solvent water may contribute, at least to a limited extent, to the rate enhancement produced by ODCase. This enzyme's active site, like that of another cofactorless enzyme recently shown to produce a rate enhancement similar in magnitude (uroporphyrinogen decarboxylase), is equipped with an ammonium group positioned in such a way as to balance the electrostatic charge of the carboxylate group of the substrate and later supply a proton to the incipient carbanion in a relatively waterless environment. PMID- 19678696 TI - Biodegradable branched polycationic polymers with varying hydrophilic spacers for nonviral gene delivery. AB - Biodegradable branched polycationic polymers with varying hydrophilic spacer lengths were synthesized from different triacrylate monomers and the amine monomer 1-(2-aminoethyl)piperazine by Michael addition polymerization. The hydrophilic spacers were varied by the number of ethyleneoxy groups in the triacrylate monomer (E/M) that ranged from 0 to 14. The polymer degradation depended on the spacer length and pH; the amount of ester degraded as determined by (1)H NMR after 14 days was 43.4 +/- 2.1% (pH 5.0) and 89.7 +/- 1.3% (pH 7.4) for the polymer with 0 E/M compared to 55.7 +/- 2.6% (pH 5.0) and 98.5 +/- 1.6% (pH 7.4) for the polymer with 14 E/M. Cell viability of rat fibroblasts after exposure to polymer solutions of concentrations up to 1000 microg/mL remained high (above 66.9 +/- 12.1% compared to below 7.6 +/- 1.1% for polyethylenimine at a concentration of 50 microg/mL or higher) and increased with the spacer length. The polyplexes made with all the synthesized polymers showed higher transfection efficiency (4.5 +/- 1.7% to 9.4 +/- 2.0%, dependent on the polymer/pDNA weight ratio) with an enhanced green fluorescent protein reporter gene compared to naked pDNA (0.8 +/- 0.4%) as quantified by flow cytometry. This study demonstrates that hydrophilic spacers can be incorporated into polycationic polymers to reduce their cytotoxicity and enhance their degradability for nonviral gene delivery. PMID- 19678699 TI - Inhibition of amyloid formation by ionic liquids: ionic liquids affecting intermediate oligomers. AB - In searching for alternative ways to reduce or inhibit amyloid formation, we have studied this process using hen egg white lysozyme (HEWL) in the presence of a low concentration of protic ionic liquids. The ionic liquids were synthesized in a combinatorial fashion maintaining the cationic part (tetramethylguanidinium) with alteration of the anionic component of each compound tested. It was observed that one of these compounds (tetramethylguanidinium acetate) inhibited amyloid formation of HEWL in vitro by nearly 50%. Examination under transmission electron microscopy confirmed the fibril inhibition, and fibrils were observed to be morphologically thinner. To investigate the mechanism of inhibition, intrinsic fluorescence, ANS binding, and circular dichroism analyses were performed. These analyses indicated that the native structure of HEWL was maintained in the presence of the ionic liquid. Performing native PAGE and nondenaturing agarose electrophoresis, it became evident that some of the intermediate oligomers were not converted to protofibrils and that the oligomers were trapped in more stable conformations. Additionally, it was observed that this inhibitory effect was related to the ionic liquid itself and not the solvated ions. It also became evident that the carboxyl functional group was important in the inhibition. The size of the anions and kosmotropicity did not play significant roles in the fibril inhibition. PMID- 19678697 TI - Insights into the mode of action of the two-peptide lantibiotic haloduracin. AB - Haloduracin, a recently discovered two-peptide lantibiotic composed of the post translationally modified peptides Halalpha and Halbeta, is shown to have high potency against a range of Gram-positive bacteria and to inhibit spore outgrowth of Bacillus anthracis. The two peptides display optimal activity in a 1:1 stoichiometry and have efficacy similar to that of the commercially used lantibiotic nisin. However, haloduracin is more stable at pH 7 than nisin. Despite significant structural differences between the two peptides of haloduracin and those of the two-peptide lantibiotic lacticin 3147, these two systems show similarities in their mode of action. Like Ltnalpha, Halalpha binds to a target on the surface of Gram-positive bacteria, and like Ltnbeta, the addition of Halbeta results in pore formation and potassium efflux. Using Halalpha mutants, its B- and C-thioether rings are shown to be important but not required for bioactivity. A similar observation was made with mutants of Glu22, a residue that is highly conserved among several lipid II-binding lantibiotics such as mersacidin. PMID- 19678698 TI - Nosiheptide biosynthesis featuring a unique indole side ring formation on the characteristic thiopeptide framework. AB - Nosiheptide (NOS), belonging to the e series of thiopeptide antibiotics that exhibit potent activity against various bacterial pathogens, bears a unique indole side ring system and regiospecific hydroxyl groups on the characteristic macrocyclic core. Here, cloning, sequencing, and characterization of the nos gene cluster from Streptomyces actuosus ATCC 25421 as a model for this series of thiopeptides has unveiled new insights into their biosynthesis. Bioinformatics based sequence analysis and in vivo investigation into the gene functions show that NOS biosynthesis shares a common strategy with recently characterized b or c series thiopeptides for forming the characteristic macrocyclic core, which features a ribosomally synthesized precursor peptide with conserved posttranslational modifications. However, it apparently proceeds via a different route for tailoring the thiopeptide framework, allowing the final product to exhibit the distinct structural characteristics of e series thiopeptides, such as the indole side ring system. Chemical complementation supports the notion that the S-adenosylmethionine-dependent protein NosL may play a central role in converting tryptophan to the key 3-methylindole moiety by an unusual carbon side chain rearrangement, most likely via a radical-initiated mechanism. Characterization of the indole side ring-opened analogue of NOS from the nosN mutant strain is consistent with the proposed methyltransferase activity of its encoded protein, shedding light into the timing of the individual steps for indole side ring biosynthesis. These results also suggest the feasibility of engineering novel thiopeptides for drug discovery by manipulating the NOS biosynthetic machinery. PMID- 19678700 TI - Production of bioactive, post-translationally modified, heterotrimeric, human recombinant type-I collagen in transgenic tobacco. AB - Collagen's biocompatibility, biodegradability and low immunogenicity render it advantageous for extensive application in pharmaceutical or biotechnological disciplines. However, typical collagen extraction from animal or cadaver sources harbors risks including allergenicity and potential sample contamination with pathogens. In this work, two human genes encoding recombinant heterotrimeric collagen type I (rhCOL1) were successfully coexpressed in tobacco plants with the human prolyl-4-hydroxylase (P4H) and lysyl hydroxylase 3 (LH3) enzymes, responsible for key posttranslational modifications of collagen. Plants coexpressing all five vacuole-targeted proteins generated intact procollagen yields of approximately 2% of the extracted total soluble proteins. Plant extracted rhCOL1 formed thermally stable triple helical structures and demonstrated biofunctionality similar to human tissue-derived collagen supporting binding and proliferation of adult peripheral blood-derived endothelial progenitor-like cells. Through a simple, safe and scalable method of rhCOL1 production and purification from tobacco plants, this work broadens the potential applications of human recombinant collagen in regenerative medicine. PMID- 19678701 TI - Intracellular Ca2+- and Mn2+-transport ATPases. PMID- 19678702 TI - De novo synthesis of modified saxitoxins for sodium ion channel study. AB - Access to novel forms of (+)-saxitoxin (STX), a potent and selective inhibitor of voltage-gated Na(+) ion channels, has been made possible through de novo synthesis. Saxitoxin is believed to lodge in the outer mouth of the channel pore, thereby stoppering ion flux. Herein, we demonstrate that modification of the C13 carbamoyl unit can be accommodated in the binding site of the protein without significantly reducing ligand-receptor affinity. These discoveries have emboldened efforts to prepare photoaffinity-labeled and other unique forms of STX as pharmacological tools for interrogating both the molecular architecture and function of Na(+) channels. A synthetic plan that makes such compounds generally available is described. PMID- 19678703 TI - Use of a high electron-affinity molybdenum dithiolene complex to p-dope hole transport layers. AB - Experimental and theoretical results are presented on the electronic structure of molybdenum tris[1,2-bis(trifluoromethyl) ethane-1,2-dithiolene] (Mo(tfd)(3)), a high electron-affinity organometallic complex that constitutes a promising candidate as a p-dopant for organic molecular semiconductors. The electron affinity of the compound, determined via inverse photoemission spectroscopy, is 5.6 eV, which is 0.4 eV larger than that of the commonly used p-dopant F(4)-TCNQ. The LUMO level of Mo(tfd)(3) is calculated to be delocalized over the whole molecule, which is expected to lead to low pinning potential. Efficient p-doping of a standard hole transport material (alpha-NPD) is demonstrated via measurements of Fermi level shifts and enhanced conductivity in alpha-NPD:1% Mo(tfd)(3). Rutherford backscattering measurements show good stability of the three-dimensional Mo(tfd)(3) molecule in the host matrix with respect to diffusion. PMID- 19678704 TI - A new reaction motif: "homo-S(N)2'-like" direct nucleophilic addition to neutral eta(3)-allylmolybdenum complexes. total synthesis of the antimalarial (+) isofebrifugine. AB - Charge neutral TpMo(CO)(2)(5-acyloxy-eta(3)-pyranyl) and TpMo(CO)(2)(5-acyloxy eta(3)-pyridinyl) scaffolds undergo a novel intermolecular "homo-S(N)2'-like" reaction with a variety of carbon nucleophiles. Combined with an annulative demetalation, the homo-S(N)2'-like substitution/annulative demetalation sequence rapidly generates 2,7-dioxabicyclo[4.3.0]nonane and 2-aza-7 oxabicyclo[4.3.0]nonane frameworks in good to excellent yields with high enantiopurity. An enantiocontrolled total synthesis of the antimalarial alkaloid (+)-isofebrifugine was achieved utilizing this reaction cascade. PMID- 19678705 TI - Asymmetric tandem wittig rearrangement/Aldol reactions. AB - A new method for the asymmetric synthesis of alpha-alkyl-alpha,beta-dihydroxy esters that involves tandem Wittig rearrangement/aldol reactions of O-benzyl- or O-allylglycolate esters derived from 2-phenylcyclohexanol is described. This sequence constructs two C-C bonds and two stereocenters, one of which is quaternary, to afford syn diol products with excellent stereocontrol. Cleavage of the chiral auxiliary affords enantiomerically enriched products with up to 95% ee. The application of this method to the preparation of a key intermediate in the synthesis of the antifungal agent alternaric acid is also described. PMID- 19678706 TI - Beneficial effects of taurine on cardiac abnormality in NZB/W F1 mice fed with a high-cholesterol diet. AB - A significantly higher prevalence of cardiovascular disease (CVD) is reported in patients with systemic lupus erythematosus (SLE) as compared with the general population and accounts for approximately 30% of deaths in SLE patients. However, the mechanism of and treatments for CVD in patients with SLE are still unclear. To explore the effects of taurine on cardiac abnormality in SLE, NZB/W F1 mice were used as the experimental model by receiving control, cholesterol, or cholesterol/taurine diets, respectively. Improved cardiac histopathological changes were observed in left ventricle tissues from the cholesterol/taurine group as compared to the control or cholesterol group. Significant reductions of TUNEL-positive cells, Fas death receptor-related components, mitochondrial dependent apoptosis, cardiac fibrosis, and fibrotic signaling components were detected in the left ventricle tissues from the cholesterol/taurine group as compared to the control or cholesterol group. Additionally, cardiac IGR1R survival signaling components were significantly increased in the left ventricle tissues from the cholesterol/taurine group as compared to the control or cholesterol group. These findings revealed the protective effects of taurine against the cardiac abnormalities in NZB/W F1 mice and may suggest the potential for clinical application of taurine in treatment of CVD in SLE. PMID- 19678707 TI - Identification of critical ligand binding determinants in Mycobacterium tuberculosis adenosine-5'-phosphosulfate reductase. AB - Mycobacterium tuberculosis adenosine-5'-phosphosulfate (APS) reductase is an iron sulfur protein and a validated target to develop new antitubercular agents, particularly for the treatment of latent infection. To facilitate the development of potent and specific inhibitors of APS reductase, we have probed the molecular determinants that underlie binding and specificity through a series of substrate and product analogues. Our study highlights the importance of specific substitutent groups for substrate binding and provides functional evidence for ligand-specific conformational states. An active site model has been developed for M. tuberculosis APS reductase that is in accord with the results presented here as well as prior structural data reported for Pseudomonas aeruginosa APS reductase and related enzymes. This model illustrates the functional features required for the interaction of APS reductase with a ligand and provides a pharmacological roadmap for the rational design of small molecules as potential inhibitors of APS reductase present in human pathogens, including M. tuberculosis. PMID- 19678708 TI - Design, synthesis, and structure-activity relationships of aminopyridine N oxides, a novel scaffold for the potent and selective inhibition of p38 mitogen activated protein kinase. AB - A novel series of aminopyridine N-oxides were designed, synthesized, and tested for their ability to inhibit p38alpha MAP kinase. Some of these compounds showed a significant reduction in the LPS-induced TNFalpha production in human whole blood. Structure-activity relationship studies revealed that N-oxide oxygen was essential for activity and was probably a determinant factor for a marked selectivity against other related kinases. Compound 45 was identified as a potent and selective p38alpha inhibitor with an appropriate balance between potency and pharmacokinetics. In vivo efficacy of 45 was demonstrated in reducing TNFalpha levels in an acute murine model of inflammation (ED(50) = 1 mg/kg in LPS-induced TNFalpha production when dosed orally 1.5 h prior to LPS administration). The oral efficacy of 45 was further demonstrated in a chronic model of adjuvant arthritis in rats with established disease when administered orally (ED(50) = 4.5 mg/kg). PMID- 19678709 TI - Serum metabolite profiling of human colorectal cancer using GC-TOFMS and UPLC QTOFMS. AB - Colorectal carcinogenesis involves the overexpression of many immediate-early response genes associated with growth and inflammation, which significantly alters downstream protein synthesis and small-molecule metabolite production. We have performed a serum metabolic analysis to test the hypothesis that the distinct metabolite profiles of malignant tumors are reflected in biofluids. In this study, we have analyzed the serum metabolites from 64 colorectal cancer (CRC) patients and 65 healthy controls using gas chromatography time-of-flight mass spectrometry (GC-TOFMS) and Acquity ultraperformance liquid chromatography quadrupole time-of-flight mass spectrometry (Acquity UPLC-QTOFMS). Orthogonal partial least-squares discriminate analysis (OPLS-DA) models generated from GC TOFMS and UPLC-QTOFMS metabolic profile data showed robust discrimination from CRC patients and healthy controls. A total of 33 differential metabolites were identified using these two analytical platforms, five of which were detected in both instruments. These metabolites potentially reveal perturbation of glycolysis, arginine and proline metabolism, fatty acid metabolism and oleamide metabolism, associated with CRC morbidity. These results suggest that serum metabolic profiling has great potential in detecting CRC and helping to understand its underlying mechanisms. PMID- 19678710 TI - The mechanism of the reaction catalyzed by uronate isomerase illustrates how an isomerase may have evolved from a hydrolase within the amidohydrolase superfamily. AB - Uronate isomerase (URI) catalyzes the reversible isomerization of D-glucuronate to D-fructuronate and of D-galacturonate to D-tagaturonate. URI is a member of the amidohydrolase superfamily (AHS), a highly divergent group of enzymes that catalyze primarily hydrolytic reactions. The chemical mechanism and active site structure of URI were investigated in an attempt to improve our understanding of how an active site template that apparently evolved to catalyze hydrolytic reactions has been reforged to catalyze an isomerization reaction. The pH-rate profiles for k(cat) and k(cat)/K(m) for URI from Escherichia coli are bell-shaped and indicate that one group must be unprotonated and another residue must be protonated for catalytic activity. Primary isotope effects on the kinetic constants with [2-2H]-D-glucuronate and the effects of changes in solvent viscosity are consistent with product release being the rate-limiting step. The X ray structure of Bh0493, a URI from Bacillus halodurans, was determined in the presence of the substrate D-glucuronate. The bound complex showed that the mononuclear metal center in the active site is ligated to the C-6 carboxylate and the C-5 hydroxyl group of the substrate. This hydroxyl group is also hydrogen bonded to Asp-355 in the same orientation as the hydroxide or water is bound in those members of the AHS that catalyze hydrolytic reactions. In addition, the C-2 and C-3 hydroxyl groups of the substrate are hydrogen bonded to Arg-357 and the carbonyl group at C-1 is hydrogen bonded to Tyr-50. A chemical mechanism is proposed that utilizes a proton transfer from C-2 of D-glucuronate to C-1 that is initiated by the combined actions of Asp-355 from the end of beta-strand 8 and the C-5 hydroxyl of the substrate that is bound to the metal ion. The formation of the proposed cis-enediol intermediate is further facilitated by the shuttling of the proton between the C-2 and C-1 oxygens by the conserved Tyr-50 and/or Arg 355. PMID- 19678711 TI - Exercise-induced bronchoconstriction in asthmatic children: a comparative systematic review of the available treatment options. AB - The aim of this article is to critically review the efficacy and safety data from randomized controlled trials (RCTs) using inhaled corticosteroids (ICSs), long- or short-acting beta(2)-adrenoceptor agonists (LABAs, SABAs), parasympatholytics and oral leukotriene receptor antagonists in the management of exercise-induced bronchoconstriction (EIB) in children with persistent asthma (EIA). The studies with sufficient information on patient characteristics and outcomes were chosen using a MEDLINE search. Results from the individual searches were combined and repeated. Studies were also found by reviewing the reference lists of the articles not included in this review. Studies focusing solely on individuals with asthma and other allergic co-morbidities (i.e. a degree of bronchial reversibility) were considered in this review. To make the paper evidence-based, the design and the quality of different studies were assessed employing the Sign criteria (evidence level [EL] and grades of recommendation [GR]). No additional statistical analyses were performed. Most of studies included paediatric patients with underlying EIA. We need to distinguish children with recurrent asthma symptoms in whom EIB is also present (patients with EIA) from asthmatic subjects whose symptoms appear only as a result of exercise (patients with EIB). Further controller treatment is indicated in patients with EIA and further reliever treatment in patients with EIB. ICSs are the first-choice controller drugs for EIA in children with persistent asthma (Sign grade of recommendation [GR]:A). In children with EIA without complete control with ICSs, SABAs (GR:A), leukotriene receptor antagonists (LTRAs) [GR:A] or LABAs (GR:A) may be added to gain control. Treatment with relievers such as SABAs (GR:A), parasympatholytics (GR:B) or, eventually, LABAs (GR:A), administered 10-15 minutes before exercise is the most preferable method of preventing EIB symptoms in children; however, not as monotherapy in children with EIA. The disadvantages and controversy relating to inhaled beta(2)-adrenoceptor agonist use lie in the development of tolerance to their effect when they are used on a regular basis, and the possibility of a resulting underuse of ICSs in patients with EIA. Researchers and guidelines recommend that if any patient requires treatment with a beta(2)-adrenoceptor agonist more than twice weekly, a low dose of ICSs should be administered. Inhaled parasympatholytics may be effective as preventive relievers in some children with EIB or EIA, especially among those with increased vagal activity. LTRAs have a well balanced efficacy-safety profile in preventing the occurrence of EIB symptoms in children. Compared with LABAs, LTRAs produce persistent attenuation of EIB and possess an additional effect with rescue SABA therapy in persistent asthmatic patients with EIA. A disadvantage of LTRAs is a non-response phenomenon. There are still insufficient data on the efficacy-safety profiles of ICS/LABA combination drugs in the treatment of EIA in children to recommend this treatment without caution. Safety profiles of inhaled SABAs, anticholinergics and montelukast in approved dosages seem sufficient enough to recommend use of these drugs in the prevention of EIB symptoms in children. Many researchers agree that treatment of EIA in children should always be individualized. PMID- 19678713 TI - Alitretinoin: in severe chronic hand eczema. AB - Alitretinoin is an endogenous retinoid and acts as a pan-agonist at retinoid receptors, binding with high affinity to both retinoic acid receptors and retinoid X receptors (RXR). Oral alitretinoin once daily is approved for use in patients with severe chronic hand eczema unresponsive to treatment with potent topical corticosteroids. In a large (n = 1032), randomized, double-blind, placebo controlled, multicentre study (BACH) of up to 24 weeks' duration in adults with severe chronic hand eczema, significantly more patients in the alitretinoin 10 or 30 mg/day groups than in the placebo group responded to treatment with clear/almost clear hands, as assessed by the Physician Global Assessment (PGA) [primary endpoint]. In an extension phase of the BACH study, alitretinoin was effective in patients who relapsed after responding to initial treatment with the drug. Of patients who had responded to initial treatment with alitretinoin 30 mg/day, significantly more alitretinoin 30 mg/day than placebo recipients responded on the PGA with clear/almost clear hands during the extension phase (primary endpoint; 80% vs 8%). Of those who had responded to initial treatment with alitretinoin 10 mg/day, 48% of alitretinoin 10 mg/day and 10% of placebo recipients responded during the extension phase. Alitretinoin was generally well tolerated in clinical trials excluding pregnant women. The most common treatment emergent adverse events and abnormal laboratory test results were consistent with those previously observed with other oral retinoids and RXR agonists. PMID- 19678714 TI - Estradiol valerate/dienogest: in oral contraception. AB - Estradiol valerate/dienogest is an oral contraceptive for women that combines the natural estrogen estradiol with the 19-nortestosterone derivative dienogest in a four-phasic formulation. Estradiol valerate/dienogest demonstrated contraceptive efficacy in a large (n = 1377), noncomparative, multicentre study in women aged 18-50 years, with 13 pregnancies over 1797.5 women-years of exposure generating an unadjusted Pearl Index (PI) of 0.73 (upper limit of 95% CI 1.24) [primary endpoint]. Six of the pregnancies were attributed to method failure, resulting in an adjusted PI, based on 1786.5 women-years of exposure, of 0.34 (upper limit of 95% CI 0.73). In a double-blind study in 798 women aged 18-50 years, estradiol valerate/dienogest and ethinylestradiol/levonorgestrel demonstrated an acceptable bleeding pattern and level of cycle control, according to several co-primary endpoints. As reported in the UK manufacturer's summary of product characteristics, the unadjusted PI for women aged 18-35 years or 18-50 years in a pooled analysis of clinical studies was 1.01 (upper limit of 95% CI 1.59) and 0.79 (upper limit of 95% CI 1.23). This pooled analysis of three studies excluded those pregnancies occurring within 14 days of the cessation of therapy. Estradiol valerate/dienogest was generally well tolerated in this population, with the nature of adverse events generally similar across the studies and between estradiol valerate/dienogest and ethinylestradiol/levonorgestrel. PMID- 19678715 TI - Dronedarone. AB - Oral dronedarone is a non-iodinated benzofurane derivative structurally related to amiodarone. Although it is considered a class III antiarrhythmic agent like amiodarone, it demonstrates multi-class electrophysiological activity. Data from the ATHENA study demonstrated that patients receiving oral dronedarone 400 mg twice daily for 12-30 months had a significantly lower risk of experiencing first hospitalization due to a cardiovascular event or death from any cause than those receiving placebo. Dronedarone exhibited rate- and rhythm-controlling properties in patients with atrial fibrilation (AF) or atrial flutter, significantly reducing the risk of a first recurrence of AF versus placebo following 12 months' therapy in the ADONIS and EURIDIS studies. In the ERATO study, dronedarone was also significantly more effective than placebo in terms of ventricular rate control. Furthermore, the beneficial effects of oral dronedarone on ventricular rate control were maintained during exercise and sustained with continued therapy. Oral dronedarone was generally well tolerated in the treatment of adult patients with AF and/or atrial flutter in clinical studies. The incidence of diarrhoea, nausea, bradycardia, rash and QT-interval prolongation was significantly higher with oral dronedarone than placebo in the large ATHENA study; however, serious cardiac-related adverse events were observed in <1% of oral dronedarone recipients. PMID- 19678716 TI - Extended-release niacin (nicotinic acid)/laropiprant. AB - Extended-release (ER) niacin (nicotinic acid)/laropiprant is a once-daily fixed dose combination tablet that has been evaluated (with or without an HMG-CoA reductase inhibitor [statin]) in the treatment of adults with dyslipidaemia or primary hypercholesterolaemia. Niacin (vitamin B3) is a lipid-modifying drug and laropiprant is an anti-flushing agent, which reduces flushing induced by niacin. In a randomized, double-blind, placebo-controlled, multicentre, 24-week trial, a significant (p < 0.001) reduction (18.4%) in plasma low-density lipoprotein cholesterol (LDL-C) levels (primary endpoint) was achieved with ER niacin/laropiprant 2000 mg/40 mg once daily (after an initial 4-week 1000 mg/20 mg once-daily regimen) compared with placebo (weeks 12-24) in adults with primary hypercholesterolaemia or mixed dyslipidaemia. ER niacin/laropiprant 2000 mg/40 mg plus simvastatin 20 mg or 40 mg once daily (after an initial 4-week lower-dose regimen) produced significant (p < 0.05) improvements, from baseline, in LDL-C levels (primary endpoint) compared with once-daily ER niacin/laropiprant 2000 mg/40 mg or simvastatin alone at week 12 in a randomized, double-blind, multicentre, factorial trial in adults with primary hypercholesterolaemia or mixed dyslipidaemia. The incidence and intensity of flushing (an efficacy endpoint) were significantly (p < 0.05) reduced with ER niacin/laropiprant compared with ER niacin in randomized trials. ER niacin/laropiprant, alone or in combination with a statin, was generally well tolerated for up to 24 weeks by adults with dyslipidaemia or primary hyercholesterolaemia. PMID- 19678717 TI - Letrozole: a review of its use in the treatment of postmenopausal women with hormone-responsive early breast cancer. AB - Letrozole (Femara) is a third-generation, nonsteroidal aromatase inhibitor. Adjuvant therapy with letrozole is more effective than tamoxifen in postmenopausal women with hormone-responsive early breast cancer, and extended adjuvant therapy with letrozole after the completion of adjuvant tamoxifen therapy is more effective than placebo in this patient population; letrozole is generally well tolerated. Ongoing trials will help answer outstanding questions regarding the optimal duration of letrozole therapy in early breast cancer and its efficacy compared with other third-generation aromatase inhibitors such as anastrozole. In the meantime, letrozole should be considered a valuable option in the treatment of postmenopausal women with hormone-responsive early breast cancer, both as adjuvant and extended adjuvant therapy. PMID- 19678719 TI - Measurements of valve circumferences and ventricular wall of fetal hearts in Chile. AB - Diagnosis of cardiac pathology, valvular stenosis, cardiac dilation, and/or cardiac hypertrophy is underestimated if measures of weight, wall thickness, and valve circumference are not obtained. Routine study methodology protocols allow us to obtain values of these measurements for a correct diagnosis of these cardiopathies. The aim of the study is to establish reference values for cardiac measurements in fetuses and to compare them with an international curve of reference. One hundred seventy-one autopsies from week 17 through 41 of gestation were performed on fetuses from the Pathology Unit at the Hospital Barros Luco in Santiago de Chile. Cases with malformations were not considered. Anthropometric and cardiac measurements for each gestational week were taken. A tabulation of data with values (in percentiles) of cardiac weight, valve circumference, and ventricular wall thickness was obtained. Values were similar to an international reference. The curves of values obtained allow for identification of normal parameters of heart weight, valve circumference, and ventricular wall thickness, thereby providing easily accessible data for each gestational week. Abnormal values that fall outside of the curve must be attributed to cardiac fetal pathology. PMID- 19678718 TI - Prasugrel: a review of its use in patients with acute coronary syndromes undergoing percutaneous coronary intervention. AB - Prasugrel (Efient) is a potent, selective and irreversible inhibitor of adenosine diphosphate (ADP)-mediated platelet aggregation that is indicated for the prevention of atherothrombotic events in patients with acute coronary syndromes (ACS) [comprising unstable angina pectoris/non-ST-segment-elevation myocardial infarction (unstable angina/NSTEMI) and ST-segment-elevation myocardial infarction (STEMI)] undergoing percutaneous coronary intervention (PCI). Oral prasugrel provides rapid, potent inhibition of platelet aggregation and is an effective antiplatelet agent for the management of patients with ACS who are undergoing PCI. In these patients, prasugrel was associated with a significantly lower incidence of ischaemic events than clopidogrel, and was particularly effective in specific subgroups of patients, such as those with diabetes mellitus. However, the efficacy of prasugrel was offset by a higher risk of bleeding than clopidogrel, with patients aged > or =75 years, those weighing <60 kg and those with a history of stroke or transient ischaemic attack at the greatest risk. A lower dose of prasugrel in patients aged > or =75 years and those weighing <60 kg may help to minimize the bleeding risk, although more data are needed to establish this; prasugrel is contraindicated in patients with a history of stroke or transient ischaemic attack. Thus, prasugrel provides a new option for the management of patients with ACS who are undergoing PCI; the risk benefit ratio should be carefully assessed before intensive antiplatelet therapy with prasugrel is initiated. PMID- 19678720 TI - AMCP Guide to Pharmaceutical Payment Methods, 2009 Update (Version 2.0). AB - The methods by which the U.S. health care system pays for prescription drugs have faced increasing scrutiny in recent years. Two key developments have emerged: (a) congressional enactment of important changes in the basis for payments for prescription drugs in the Medicare and Medicaid programs; and (b) a March 2009 decision in a federal class action lawsuit that alleged fraudulent manipulation of the dominant pricing benchmark (average wholesale price, AWP), used primarily as the basis for payment for brand-name prescription drugs. The debate about prescription drug payment methods centers on determining the most appropriate basis for calculating how payers, including patients, government agencies, employers, and health plans, should pay pharmacies and other providers for drugs. Historically, payment for prescription drugs has been based on published prices that do not necessarily reflect the actual acquisition costs paid by providers, primarily pharmacies, physicians, and hospitals. This has led policymakers to believe that Medicare and Medicaid programs have paid more than is necessary for prescription drugs. Thus, in an effort to reform the payment system and reduce drug expenditures, policymakers have made significant changes to the benchmarks used by public programs to pay for drugs, and in some instances have created new benchmarks. Private payers have followed the government's lead and begun to change their own payment methods and benchmarks. They can be expected to accelerate the change as a result of the settlement agreement approved in the March 2009 federal court decision. The settlement will result in the lowering of the AWP for more than 400 generic and brand-name drugs. In addition - and technically unrelated to the litigation and any appeals that may be taken - 2 major price data reporting companies, First DataBank and Medi-Span, announced their intent to discontinue publication of AWP within 2 years of September 26, 2009. (At the time this report was prepared, there have been no similar announcements from Thomson Healthcare for Redbook or from Elsevier for Gold Standard [ProspectoRx], who are 2 other publishers of prescription drug prices). Furthermore, several manufacturers have announced that they will no longer provide either an AWP or a markup percentage on certain pharmaceuticals.3 Thus, by 2011, the AWP benchmark as we know it will no longer be widely available for use by public or commercial payers for payment of pharmaceutical products. PMID- 19678721 TI - Clinical, economic, and quality of life impact of atrial fibrillation. AB - BACKGROUND: Atrial fibrillation (AF) is a common, age-related arrhythmia that disproportionately affects men, adversely affects quality of life, and causes considerable morbidity and mortality. OBJECTIVES: To describe trends in the prevalence and incidence of AF in the United States; discuss the etiologies and complications of AF; characterize the economic burden of AF; and predict an individual's risk for developing AF and AF-related stroke. SUMMARY: The prevalence and incidence of AF in the United States are expected to increase in the coming decades because of the aging of the population; improved survival rates associated with coronary heart disease, heart failure, and hypertension; and increased rate of performance of surgical procedures. The economic burden of AF is substantial because of high rates of hospitalization and other health resource utilization. Hypertension, coronary heart disease, and systolic heart failure are the most important risk factors for AF. Ischemic stroke is the most devastating complication of AF. Risk factors for stroke in patients with AF include recent congestive heart failure, hypertension, advanced age, diabetes mellitus, and a history of stroke or transient ischemic attack. Risk scoring systems have been developed to predict an individual's risk for developing AF and the risk for stroke in a patient with AF. The estimated lifetime risk for AF in men and women aged 40 years of age or older is 1 in 4, which is higher than the risk for other diseases that are a common cause for concern among elderly patients. CONCLUSIONS: The clinical and economic burden of AF in the United States is large and will continue to increase in the future. The use of scoring systems to predict the risk of AF and AF-related stroke affords clinicians the opportunity to intervene to minimize these risks and improve patient outcomes. PMID- 19678722 TI - Pharmacologic management of atrial fibrillation: established and emerging options. AB - BACKGROUND: In patients with atrial fibrillation (AF), antiarrhythmic drug therapy currently plays a greater role in maintaining sinus rhythm after cardioversion than it does in converting AF to sinus rhythm. Amiodarone is the most effective antiarrhythmic agent for maintaining sinus rhythm after cardioversion in patients with AF. However, its pharmacokinetics is complex; the drug interacts with many commonly used medications; and long-term use can cause thyroid dysfunction, hepatotoxicity, and other severe extracardiac adverse effects. The use of antiarrhythmic strategies in patients with AF has decreased because of evidence of greater safety and lower costs for hospitalization obtained from the use of rate-control strategies instead. Nevertheless, some patients require a rhythm-control strategy. Warfarin is used to prevent embolic stroke in many patients with AF, but its use is also complex and requires monitoring. Therefore, efforts have been made to develop antiarrhythmic agents with improved tolerability and anticoagulants that are easy to use. OBJECTIVES: To describe the 3 primary goals of pharmacotherapy in patients with AF, compare and contrast the efficacy and safety of established and investigational pharmacotherapies for AF, and recommend a drug regimen for an individual with AF based on patient-specific factors. SUMMARY: Currently available antiarrhythmic agents differ in their efficacy for maintaining sinus rhythm after cardioversion in AF patients with tolerability problems, comorbidities (particularly heart failure and renal impairment), and potential drug interactions. Hence, when selecting drug therapy to maintain sinus rhythm after cardioversion, it is important to take into consideration patient characteristics, including age, disease states, renal function, and concurrent drug therapies. Outpatient self administration of single loading doses of flecainide or propafenone with what is referred to as the pill-in-the-pocket approach may be considered for carefully selected patients with recurrent episodes of symptomatic AF. The recently approved antiarrhythmic agent dronedarone has electrophysiologic properties similar to those of amiodarone, but its lack of iodine may improve upon the pharmacokinetic and tolerability issues associated with amiodarone. Vernakalant is another investigational antiarrhythmic agent that may prove useful for cardioversion and maintenance of sinus rhythm after cardioversion in patients with AF. New oral anticoagulants that do not require close laboratory monitoring and are simpler to use than warfarin have been used investigationally for prevention of venous thromboembolism and are in clinical trials for prevention of embolic stroke in patients with AF. CONCLUSIONS: Pharmacotherapy for patients with AF should be individualized based on patient-specific factors. New therapeutic options may become available to facilitate treatment of these patients. PMID- 19678723 TI - Improving cost-effectiveness of and outcomes from drug therapy in patients with atrial fibrillation in managed care: role of the pharmacist. AB - BACKGROUND: The medical care costs for procedures, medications, and testing associated with atrial fibrillation (AF) in the United States are high and projected to increase markedly in the future as the number of Americans affected grows. The burden on patient quality of life, the health care system, and society are pharmacoeconomic considerations in managing AF. OBJECTIVES: To identify key pharmacoeconomic considerations in managing AF and describe ways in which managed care pharmacists can improve the cost-effectiveness of and outcomes from drug therapy for AF. SUMMARY: The high medical care costs of AF are largely the result of the high cost of hospitalization and inpatient procedures. Recurrence of AF dramatically increases costs, especially for hospital care. Managed care pharmacists have many opportunities to provide cost-effective care to and improve outcomes in patients with AF. Policy and process review, population management, and case management are key strategies for improving outcomes in patients with AF. Pharmacist input into policy and process review, including pharmacy benefits design, formulary management, and the use of information technology, can help ensure that the use of drug therapy for AF is cost-effective. Population management strategies, such as development of clinical pathways and patient registries, seek to improve the quality, consistency, and cost-effectiveness of care and the likelihood that desired therapeutic outcomes are achieved through targeted interventions. Case management strategies focus on longitudinal care for individuals in order to improve quality. Pharmacist-managed anticoagulation services and antiarrhythmic drug monitoring are the 2 most widely known case management strategies for patients with AF. Managed care pharmacists can screen patients with AF for the use of anticoagulation, which is needed to prevent embolic stroke but is under-used, even though recommended by evidence-based guidelines. The clinical efficacy and cost-effectiveness of pharmacist-managed anticoagulation services for patients with AF are well documented. Pharmacist managed antiarrhythmic drug monitoring is a less well-known case management strategy that facilitates early detection and intervention to minimize toxicity. CONCLUSIONS: Managed care pharmacists can play an instrumental role in implementing strategies to improve the cost-effectiveness of and outcomes from drug therapy for AF. PMID- 19678724 TI - Animal experimentation and the Three Rs: time for honest answers to some leading questions. PMID- 19678725 TI - The evolution of the Three Rs. AB - Whilst the whole world is celebrating the bicentenary of the birth of Charles Darwin and the 150th anniversary of the publication of his renowned book, The Origin of Species, another anniversary should not be forgotten - the publication of The Principles of Humane Experimental Technique by W.M.S. Russell and R.L. Burch. The concomitance of the anniversaries of the two publications is not a coincidence, since, as reflected by the numerous quotes chosen by Russell from Darwin's masterpiece, numerous analogies can be found between the two works and the new ideas they describe. From a discrete birth, and after decades of struggle, the Three Rs concept can now celebrate its 50th anniversary, the result of its evolution through harsh selection and adaptation. The emergence of new types of techniques, in combination with the descent of modified old ones, testify to the undeniable change in our society toward a more efficient and more ethical science, through the progressive replacement of animal models. Both Darwin and Russell would no doubt have welcomed such progress, not only in terms of science, but also of moral values. One could also expect that, if Russell could have foreseen the incredible technological advances achieved 50 years later, where Replacement becomes a reality, as illustrated by some edifying examples, The Principles of Humane Experimental Technique would have probably been defined as the One R concept. PMID- 19678727 TI - Prefatory Note. PMID- 19678726 TI - The origins and early days of the Three Rs concept. AB - Some questions are answered concerning the origins of the Three Rs (Replacement, Reduction, Refinement) concept in relation to animal experimentation, expounded 50 years ago by Russell and Burch in The Principles of Humane Experimental Technique, by reference to some key publications and to correspondence in the W.M.S. and Claire Russell Archive, which is currently being established at the University of Nottingham. Some insight is also given into the relationship between Russell and Burch, the first use of "alternatives" in the Three Rs context, and the background to the publication of the book. PMID- 19678728 TI - The progress of humane experimental technique since 1959: a personal view. AB - This is very much a personal interpretation of the Three Rs and the efforts by scientists connected with the more humane use of animals. It is intended to illustrate that scientists are given a job to do and, if it involves animals, they have little choice but to use them. It is also intended to illustrate that so many of those so involved are the very people who have made gigantic efforts in finding ways of replacing, reducing and/or refining techniques which require animals. PMID- 19678729 TI - The progress of humane experimental technique: the first annual FRAME lecture. PMID- 19678730 TI - Ethics: views from IACUC members. AB - Institutional Animal Care and Use Committee (IACUC) members were interviewed on various ethical matters, including ethics, animal ethics, science and ethics, and the use of animals in research, in order to explore their implicit ethical framework. The results revealed that IACUC members entertain rich and diverse beliefs about ethics, that are part of an implicit ethical framework which relates to different domains of knowledge, such as biology (differences between human and animals), psychology (e.g. affective relationships with pets), and so on. The results also revealed that IACUC members hold quite a restrictive view on both animal ethics and animal use in research, and that they apply implicit ethical notions, such as respect and justice, to some elements (e.g. ethical rules) of the explicit ethical framework they are provided with when performing ethical evaluations of animal use. The study suggests that IACUC members should be provided with more up-to-date information on topics such as animal ethics and animal use in research. PMID- 19678731 TI - Assessing the search for and implementation of the Three Rs: a survey among scientists. AB - A survey among scientists into the current practice of searching for Replacement, Reduction and Refinement (Three Rs) alternatives, highlights the gap between the statutory required need to apply the Three Rs concept whenever possible and the lack of criteria for searching for Three Rs alternatives. A questionnaire was distributed to 342 scientists (Federation of European Laboratory Animal Science Associations [FELASA] Category C and B individuals), of which 67 responded. These scientists are customers of the Central Animal Laboratory of Radboud University Nijmegen Medical Centre. The results indicate that there is room for improvement in searching effectively for the Three Rs: skills in searching biomedical databases for Three Rs alternatives are limited, knowledge of specialised Three Rs databases is very limited, and satisfaction on the availability and accessibility of Three Rs information is low. None of the respondents allocate budget for a specific Three Rs alternatives search, although 50% do spend, on average, two hours engaged in this search for each application to their animal ethics committees. The majority of the respondents expressed the wish that the search for alternatives could be easier and less time consuming, and prefer to achieve this through the service offered by specialists at the Central Animal Laboratory. On the basis of the results from the questionnaire, the 3R Research Centre was established, with the aim of providing services and support for biomedical scientists, to improve the search for, and subsequent implementation of, the Three Rs. PMID- 19678732 TI - Selection of chemicals for the development and evaluation of in vitro methods for skin sensitisation testing. PMID- 19678733 TI - A review of the implementation of the embryonic stem cell test (EST). The report and recommendations of an ECVAM/ReProTect Workshop. PMID- 19678734 TI - Early enteral feeding in conservatively managed stage II necrotizing enterocolitis is associated with a reduced risk of catheter-related sepsis. AB - AIMS: To compare the effect of fasting period duration on complication rates in neonates managed conservatively for necrotizing enterocolitis (NEC) Bell stage II. METHODS: We conducted a multicenter study to analyze retrospectively multiple data collected by standardized questionnaire on all admissions for NEC between January 2000 and December 2006. NEC was staged using modified Bell criteria. We divided the conservatively managed neonates with NEC Bell stage II into two groups (those fasted for <5 days and those fasted for >5 days) and compared the complication rates. RESULTS: Of the 47 conservatively managed neonates Bell stage II, 30 (64%) fasted for <5 days (range 1-4 days) and 17 (36%) for >5 days (range 6-16 days). There were no significant differences for any of the patient characteristics analyzed. One (3%) and four (24%) neonates, respectively, developed post-NEC bowel stricture. One (3%) and two neonates (12%) suffered NEC relapse. None and five (29%) neonates developed catheter-related sepsis. CONCLUSION: Shorter fasting after NEC appears to lower morbidity after the acute phase of the disease. In particular, shorter-fasted neonates have significantly less catheter-related sepsis. We found no benefit in longer fasting. PMID- 19678735 TI - The RET functional variant c 587T>C is not associated with susceptibility to sporadic medullary thyroid cancer. PMID- 19678736 TI - In vitro studies on the relationship between the anti-inflammatory activity of Physalis peruviana extracts and the phagocytic process. AB - The study of plants used in traditional medicine has drawn the attention of researchers as an alternative in the development of new therapeutics agents, such as the American Solanaceae Physalis peruviana, which has significant anti inflammatory activity. The Physalis peruviana anti-inflammatory effect of ethanol or ether calyces extracts on the phagocytic process was assessed by using an in vitro phagocytosis model (Leishmania panamensis infection to murine macrophages). The Physalis peruviana extracts do not inhibit microorganism internalization and have no parasiticide effect. Most ET and EP extracts negatively affected the parasite's invasion of macrophages (Infected cells increased.). This observation might result from a down-regulation of the macrophage's microbicide ability associated with a selective reduction of proinflammatory cytokines levels. Physalis peruviana's anti-inflammatory activity described in this model is related to an immunomodulatory effect exerted on macrophages infected, which directly or indirectly "blocks" their ability to secrete soluble proinflammatory mediators. PMID- 19678737 TI - Thyroid function in Japanese adults as assessed by a general health checkup system in relation with thyroid-related antibodies and other clinical parameters. AB - BACKGROUND: A limited number of epidemiological studies have attempted to assess thyroid function in the general population of iodine-sufficient countries. The aim of the present study was to determine the underlying thyroid diseases responsible for abnormal thyroid function detected by a general health checkup system in Japan, and to characterize the lipid metabolism in subjects found to have thyroid dysfunction. METHODS: Serum thyrotropin (TSH), free thyroxine, anti thyroglobulin antibodies (TgAb), anti-thyroid peroxidase antibodies (TPOAb), and TSH-binding inhibitor immunoglobulins (TBII) were determined in 1818 Japanese adults (804 men and 1014 women; mean age 51.3 +/- 9.0 years) who undertook a general health checkup. RESULTS: Of the 1818 examinees, 12 (0.7%) had overt hypothyroidism (OH), 105 (5.8%) subclinical hypothyroidism, 13 (0.7%) overt thyrotoxicosis, and 39 (2.1%) subclinical thyrotoxicosis. TgAb or TPOAb tests were positive in 17.7% of men and 31.4% of women. The prevalence of positive tests for TgAb or TPOAb was 14.8% for men and 23.4% for women without palpable goiter. Positive tests for TgAb, TPOAb, TBII, and a palpable goiter were more common in subjects with abnormal thyroid function tests than in subjects with normal thyroid function. At the time that abnormal thyroid function test results were first obtained, the signs of thyrotoxicosis were mild or even absent in all 13 subjects with overt thyrotoxicosis, 8 of whom had Graves' disease and 5 of whom had painless thyroiditis. Of the 12 patients with OH, only 2 patients had a palpable goiter. In the OH group, TgAb tests were positive in eight, TPOAb tests were positive in eight, and TBII tests were positive in two. The prevalence of disturbed lipid metabolism, when adjusted for age, was significantly higher in the subclinical hypothyroidism group than in normal controls (p < 0.001; odds ratio, 1.67; 95% confidence interval, 1.10-2.51). CONCLUSIONS: In Japanese adults who chose to be screened by a general health checkup system, the prevalence of abnormal thyroid function was nearly 10%. In a high percentage of these patients, abnormal thyroid function could not be detected by their history or physical examination. Just a physical examination without thyroid function tests, particularly serum TSH levels, was not adequate even when performed by a thyroid specialist. PMID- 19678738 TI - Calcitonin estimation in neck lymph node fine-needle aspirate fluid prevents misinterpretation of cytology in patients with metastatic medullary thyroid cancer. PMID- 19678712 TI - Efflux-mediated drug resistance in bacteria: an update. AB - Drug efflux pumps play a key role in drug resistance and also serve other functions in bacteria. There has been a growing list of multidrug and drug specific efflux pumps characterized from bacteria of human, animal, plant and environmental origins. These pumps are mostly encoded on the chromosome, although they can also be plasmid-encoded. A previous article in this journal provided a comprehensive review regarding efflux-mediated drug resistance in bacteria. In the past 5 years, significant progress has been achieved in further understanding of drug resistance-related efflux transporters and this review focuses on the latest studies in this field since 2003. This has been demonstrated in multiple aspects that include but are not limited to: further molecular and biochemical characterization of the known drug efflux pumps and identification of novel drug efflux pumps; structural elucidation of the transport mechanisms of drug transporters; regulatory mechanisms of drug efflux pumps; determining the role of the drug efflux pumps in other functions such as stress responses, virulence and cell communication; and development of efflux pump inhibitors. Overall, the multifaceted implications of drug efflux transporters warrant novel strategies to combat multidrug resistance in bacteria. PMID- 19678739 TI - Ammonium sulfate gradient loading of brucine into liposomes: effect of phospholipid composition on entrapment efficiency and physicochemical properties in vitro. AB - BACKGROUND: Brucine, the major active alkaloid constituent extracted from traditional Chinese herbal medicine Nux vomica, had been found to possess remarkable antitumor, analgesic, and anti-inflammatory activities. In this study, we attempted to encapsulate brucine into liposomes to improve its therapeutic effects. The entrapment efficiency (EE) and the stability of liposomes are two key factors associated with the therapeutic effects of liposomal drugs. We developed a novel liposome-based brucine formulation that was composed of soybean phosphatidylcholine (SPC) and hydrogenated soybean phosphatidylcholine (HSPC). METHOD: The liposomes with different phospholipid composition were characterized for their EE, vesicle size, drug release profile, and leakage in vitro. RESULTS: The molar ratio of HSPC/SPC = 1:9 was determined as the optimum ratio. Compared with conventional liposomes composed of only SPC or HSPC, EE of the brucine loaded novel liposomes was increased markedly, especially at high drug/lipid molar ratios. The results of drug release showed that the novel liposomes were more stable than the conventional SPC liposomes in the presence of fetal calf serum. In addition, the results of the leakage experiments revealed that the novel liposomes also had better stability in phosphate buffer solution (PBS) with respect to drug retention. Although the conventional HSPC liposomes is more stable than the novel liposomes, the novel liposomes composed of 10% HSPC and 90% SPC may still have promising application potential because HSPC is much more expensive than SPC. CONCLUSION: Taken together, efficient encapsulation of brucine into the novel liposomes, their improved stability, and the price of phospholipids indicate that the novel liposomes may act as promising carriers for active alkaloids such as brucine. PMID- 19678740 TI - Indicators of compliance for developmental follow-up of infants discharged from a regional NICU. AB - AIM: To identify factors associated with compliance of scheduled outpatient developmental follow-up appointments in an effort to better ensure future care. METHODS: This retrospective observational cohort study looked at patients born between January 7(th) 2006 and June 30(th) 2007 and discharged from a regional neonatal intensive care unit (RNICU). Discharge summaries were reviewed to attain information regarding 16 patient descriptives and 12 patient morbidities. Data were recorded and analyzed utilizing the statistical software SPSS 11.5. RESULTS: Children of older mothers were more likely to attend follow-up (compliant: 30 years vs. non-compliant: 27 years). Factors which significantly improved compliance with follow-up care were patient contact after discharge (compliant: 65% vs. non-compliant: 35%) and early intervention referral (compliant: 64% vs. non-compliant: 36%). Factors which significantly hindered compliance were maternal drug use during pregnancy (compliant: 11.8% vs. non-compliant: 88%), and patient transfer to outside NICUs [(transferred out: compliant: 3 (10.3%), non compliant 25 (89.3%)]. CONCLUSIONS: Several factors associated with compliance have been identified. Direct patient contact after discharge positively correlated with improved follow-up attendance. The severity of patient disease in the NICU did not impact follow-up rates. As a result close attention needs to be paid to factors which influence compliance with outpatient follow-up for developmental screening. PMID- 19678742 TI - Utility of two-dimensional echocardiography in pregnancy and post-partum period and impact on management in an inner city hospital. AB - AIM: Evaluate the practice and appropriateness of requesting echocardiograms in patients with suspected or documented cardiac disease during gestation and puerperium, using the American College of Cardiology Foundation (ACCF) appropriateness criteria, in conjunction with clinical picture. METHODS: Retrospective observational study, to analyze echocardiograms performed during pregnancy and puerperium at a teaching hospital from 2001 to 2006 for appropriateness criteria and studying its impact on management. Sixty-seven patients pregnant or in the puerperal stage had an echocardiogram performed during that period; 58 met our criteria for inclusion. RESULTS: Based on clinical information and criteria of the ACCF, 51 of the 58 echocardiograms met the appropriateness criteria. Of the 51, results of 40 impacted on management; 14 of the 40 echocardiograms that had an impact were abnormal. CONCLUSIONS: Although the ACCF appropriateness criteria have not been specifically studied in pregnancy, our study demonstrates that the criteria are applicable if used appropriately in pregnancy. Most indications in our study correlated with the appropriateness criteria. Although most findings were normal, information from echocardiograms impacted on management in the majority of patients, contributing to therapeutic decision-making. The reliability of echocardiograms performed according to appropriate criteria to assist clinical decisions was excellent even in patients with physiologic cardiovascular changes. PMID- 19678743 TI - Takayasu's arteritis in pregnancy: review of literature and discussion. AB - Abstract Takayasu's arteritis (TA) is a rare inflammatory disease of the arteries that affects women of childbearing age. The optimal management for pregnant patients with this disease has not yet been defined. The course of disease seems to be neither affected nor worsened by pregnancy. We could not find reported maternal deaths directly related to pregnancy. However, many authors report maternal as well as fetal unfavorable events in the course of pregnancy. We describe a 25-year-old primigravida of Turkish-Greek origin who presented at 30 weeks of pregnancy with active TA. In the 37(th) week, intrauterine fetal death occurred. Our patient did not show high blood pressure or aortic inflammation. The course of her disease was stable. Whether a newly diagnosed TA during pregnancy should be regarded as an indication for premature delivery is discussed. An interdisciplinary collaboration of rheumatologists, nephrologists and obstetricians is necessary to improve maternal and fetal prognosis. PMID- 19678741 TI - Dual functions of thyroid hormone receptors in vertebrate development: the roles of histone-modifying cofactor complexes. AB - Thyroid hormone (TH) receptor (TR) plays critical roles in vertebrate development. Transcription studies have shown that TR activates or represses TH inducible genes by recruiting coactivators or corepressors in the presence or absence of TH, respectively. However, the developmental roles of these TR cofactors remain largely unexplored. Frog metamorphosis is totally dependent on TH and mimics the postembryonic period in mammalian development during which TH levels are also high. We have previously proposed a dual function model for TR in the development of the anuran Xenopus laevis. That is, unliganded TR recruits corepressors to TH-inducible genes in premetamorphic tadpoles to repress these genes and prevent premature metamorphic changes and subsequently, when TH becomes available, liganded TR recruits coactivators to activate these same genes, leading to metamorphosis. Over the years, we and others have used molecular and genetic approaches to demonstrate the importance of the dual functions of TR in Xenopus laevis. In particular, unliganded TR has been shown to recruit histone deacetylase-containing corepressor complexes in premetamorphic tadpoles to control metamorphic timing. In contrast, metamorphosis requires TH-bound TR to recruit coactivator complexes containing histone acetyltransferases and methyltransferases to activate transcription. Furthermore, the concentrations of coactivators appear to regulate the rate of metamorphic progression. Studies in mammals also suggest that the dual function model for TR is conserved across vertebrates. PMID- 19678744 TI - Painless thyroiditis and radioactive iodine therapy. PMID- 19678745 TI - High prevalence of side effects after recombinant human thyrotropin-stimulated radioiodine treatment with 30 mCi in patients with multinodular goiter and subclinical/clinical hyperthyroidism. AB - BACKGROUND: Treatment of multinodular goiters (MNGs) is highly controversial. Radioiodine (RAI) therapy is a nonsurgical alternative for the elderly who decline surgery. Recently, recombinant human thyrotropin (rhTSH) has been used to augment RAI uptake and distribution. In this study, we determined the outcome of 30 mCi RAI preceded by rhTSH (0.1 mg) in euthyroid (EU) and hyperthyroid (subclinical/clinical) patients with large MNGs. METHODS: This was a prospective cohort study. Forty-two patients (age, 43-80 years) with MNGs were treated with 30 mCi RAI after stimulation with 0.1 mg of rhTSH. Patients were divided into three groups, according to thyroid function: EU (n = 18), subclinically hyperthyroid (SC-H, n = 18), and clinically hyperthyroid (C-H, n = 6). All patients underwent a 90-day low-iodine diet before treatment, and those with clinical hyperthyroidism received methimazole 10 mg daily for 30 days. Serum TSH, free thyroxine (FT4), total triiodothyronine (TT3), and thyroglobulin were measured at baseline and at 24, 48, 72, 168 hours, and 1, 3, 6, 9, 12, 18, 24, and 36 months after therapy. Thyroid volume was assessed by computed tomography at baseline and every 6 months. RESULTS: Patients had high iodine urinary excretion (308 +/- 108 microg I/L) at baseline. TSH levels at baseline were within the normal range (1.5 +/- 0.7 microU/mL) in the EU group and suppressed (<0.3 microU/mL) in the SC-H and C-H groups. After rhTSH, serum TSH peaked at 24 hours reaching 12.4 +/- 5.85 microU/mL. After RAI administration, patients in both hyperthyroid groups had a higher increase in FT4 and TT3 compared with those in the EU group (p < 0.001). Thyroglobulin levels increased equally in all three groups until day 7. Thyroid volume decreased significantly in all patients. Side effects were more common in the SC-H and C-H groups (31.4% and 60.4%, respectively) compared with EU patients (17.8%). Permanent hypothyroidism was more prevalent in the EU group (50%) compared with the SC-H (11%) and C-H (16.6%) groups. CONCLUSIONS: Patients with MNG may have subclinical and clinical nonautoimmune iodine-induced hyperthyroidism. Despite a low-iodine diet and therapy with methimazole, hyperthyroid patients have a significantly higher increase in FT4 and TT3 levels after RAI ablation. This can lead to important side effects related mostly to the cardiac system. We strongly advise that patients with SC-H and C-H be adequately treated with methimazole and low-iodine diet aiming to normalize their hyperthyroid condition before rhTSH-stimulated treatment with RAI. PMID- 19678746 TI - Results of rosiglitazone therapy in patients with thyroglobulin-positive and radioiodine-negative advanced differentiated thyroid cancer. AB - BACKGROUND: Rosiglitazone is a peroxisome proliferator-activated receptor (PPAR) gamma agonist that has shown promise as both an antiproliferative and redifferentiating agent for the treatment of thyroid cancer in preclinical studies. We investigated the efficacy and side effects of rosiglitazone therapy in patients with differentiated thyroid cancer of follicular cell origin that fails to take up radioiodine or is unresectable. METHODS: Twenty patients with differentiated thyroid cancer were enrolled in an open-label, phase II trial of oral rosiglitazone treatment (4 mg daily for 1 week, then 8 mg daily for 7 weeks). RESULTS: Five of 20 patients had a positive radioiodine scan after rosiglitazone treatment. Four patients had radioiodine uptake in the neck and one patient had uptake in the pelvis. Unstimulated thyroglobulin levels after rosiglitazone treatment increased in five patients, remained stable in 12 patients, and decreased in three patients. Seven patients had progressive disease on follow-up cross-sectional imaging; six patients in the size and number of lung metastasis and two patients in the size of the neck tumors. Overall, five patients had a partial response (decreased thyroglobulin or positive radioiodine uptake), three patients had stable disease (no change in thyroglobulin and radioiodine uptake status), and 12 patients had disease progression (increased thyroglobulin). By RECIST criteria, no patient had a complete or partial response to rosiglitazone treatment at 3 months follow-up. The mean follow-up time after protocol treatment was 12 months (median 12 months). CONCLUSIONS: Our findings suggest that rosiglitazone therapy may induce radioiodine uptake and reduce serum thyroglobulin levels in some patients with differentiated thyroid cancer but this did not result in clinically significant response on long-term follow-up. Moreover, no patients had response to rosiglitazone therapy by anatomic imaging studies. PMID- 19678747 TI - Thyroid cellular models: a warning concerning their application. PMID- 19678748 TI - Use of supplements with and without iodine in women of childbearing age in the United States. PMID- 19678749 TI - Influence of the coating formulation on enzymatic digestibility and drug release from 5-aminosalicylic acid pellets coated with mixtures of high-amylose starch and Surelease intended for colon-specific drug delivery. AB - BACKGROUND: Colon-specific delivery of drugs can be achieved with dosage forms coated with biopolymers that are metabolized selectively by the colonic microflora and yet resistant to enzymatic digestion in the small intestine. AIM: The aim of this study was to study the influence of formulation factors on the performance of mixed films from high-amylose starches and Surelease((R)), applied using a spray-coating process, as potential colon-specific delivery devices. METHODS: 5-Aminosalicylic acid-loaded pellets were prepared by an extrusion spheronization process and film coated with mixtures of the starches and Surelease((R)). Optimization of the coating formulation, that is, starch-to Surelease((R)) ratio, film-coating thickness, and type of starch, was undertaken first in enzyme-free media resembling the conditions in the stomach and small intestine. The effect of curing of the film coating on the drug release profile upon storage was also evaluated. Optimized coating formulations were further assessed for enzymatic digestibility using artificial gastric and intestinal juices containing commercially available pepsin and pancreatin or alpha-amylase from hog pancreas, respectively. Finally, drug release was assessed in fluid simulating conditions in the colon (SCF) containing Bacillus licheniformis alpha amylase. RESULTS: Film coatings comprising high-amylose starches and Surelease((R)) in a ratio of 1:2 (w/w) and film thickness of approximately 45 microm were able to withstand the chemical and enzymatic environment of the upper gastrointestinal tract, in particular, resisted degradation by the pancreatic alpha-amylases. Stability of the coatings during storage was achieved with additional curing. In SCF, these coatings were susceptible to enzymatic degradation. CONCLUSIONS: This study showed that high amylose starch-mixed films can be successfully used as colon-specific delivery devices. The preparation of the coating dispersions described is simple and rapid, without the need to extract the amylose component of starch. PMID- 19678750 TI - Histopathologic diagnosis of thyroid tuberculosis. AB - BACKGROUND: Thyroid tuberculosis is rarely encountered. Due to the absence of specific signs and symptoms, the diagnosis is difficult without histopathologic examination of surgical material. In this study, we aim to present histopathologic findings of our patients with thyroid tuberculosis. METHODS: We retrospectively reviewed 800 thyroidectomy specimens from the pathology laboratories of two medical centers that were obtained over a 5-year period. We reviewed clinical and laboratory data, fine-needle aspiration cytology smears, and 4-microm formalin-fixed paraffin-embedded hematoxylin-eosin sections and Erlich Ziehl Nelsen-stained sections of granulomatous thyroiditis. RESULTS: We found nine cases of granulomatous thyroiditis and detected five cases of thyroid tuberculosis. Thyroid ultrasound revealed hypoechoic nodules ranging between 1.3 and 2.5 cm in all patients with thyroid tuberculosis. Fine-needle aspiration cytology smears, which showed thyrocytes, macrophages, and lymphocytes with a colloid background, were nonspecific. Microscopic examination of all thyroids revealed granulomas of different sizes, which were composed of Langhans' giant cells, epithelioid histiocytes, and lymphocytes around caseation necrosis. In Erlich Ziehl Nelsen-stained sections of three cases, bacilli were seen. Bacilli were identified in the microbiology culture of the remaining two patients. CONCLUSIONS: Thyroid tuberculosis is diagnosed by histopathologic examinations and microbiologic cultures. When granulomatous thyroiditis is encountered, tuberculosis should be kept in mind and Erlich Ziehl Nelsen staining should be performed for prompt diagnosis. PMID- 19678751 TI - Platelet inhibitory effects of OTC doses of naproxen sodium compared with prescription dose naproxen sodium and low-dose aspirin. AB - OBJECTIVES: Prescription dose naproxen has been reported to have an antiplatelet effect similar to low-dose aspirin (ASA). This study evaluated the platelet inhibitory effects of over-the-counter (OTC) doses of naproxen sodium (NAPSO) compared to that of a prescription dose of NAPSO and to low-dose enteric-coated aspirin (EC-ASA). RESEARCH DESIGN AND METHODS: This was a phase I, open-label, randomized, placebo-controlled, two-way crossover, multi-dose, pharmacodynamic trial conducted in healthy male and female volunteers (n = 48, mean age = 41.7 years). All subjects received 7 days of either prescription dose NAPSO (550 mg twice daily), OTC doses of NAPSO (220 mg two or three times daily), or placebo twice daily (period 1). After a minimum 6-day washout period, all subjects then received 7 days of EC-ASA 81 mg once daily (period 2). All study medications were taken by mouth. PRIMARY OUTCOME MEASURE: Inhibition of serum thromboxane B(2) (TXB(2)), as a marker of platelet cyclooxygenase-1 (COX-1) inhibition, measured 24 h after the day 7 morning dose. This was measured after both period 1 and period 2. RESULTS: After 7 days of treatment in period 1, mean inhibition of TXB(2) was 47% for placebo and > or =98% for all doses of NAPSO. After 7 days of EC-ASA 81 mg, mean inhibition of TXB(2) was > or = 97% (period 2). STUDY LIMITATIONS: Out-patient study setting. CONCLUSIONS: These data suggest that OTC doses of NAPSO (220 mg two or three times daily) have an antiplatelet effect similar to EC-ASA 81 mg and to prescription dose NAPSO (550 mg twice daily). PMID- 19678752 TI - Evidence for moxifloxacin in community-acquired pneumonia: the impact of pharmaco economic considerations on guidelines. AB - OBJECTIVE: In an era of limited resources, policy makers and health care payers are concerned about the costs of treatment in addition to its effectiveness. However, guidelines do not tend to consider the cost-effectiveness of treatment options. This paper aims to conduct an international literature review with a view to assessing the impact of pharmaco-economic considerations of CAP treatment with moxifloxacin on recent guidelines. METHODS: The pharmaco-economic state of the art of treating CAP with moxifloxacin is assessed and compared with guidelines issued by the European Respiratory Society and by the Infectious Diseases Society of America/American Thoracic Society. Also, evidence on moxifloxacin consumption and antimicrobial resistance, and the impact of resistance on the cost-effectiveness of moxifloxacin is reviewed. Studies were identified by searching PubMed, Centre for Reviews and Dissemination databases, Cochrane Database of Systematic Reviews, and EconLit up to January 2009. RESULTS: The existing pharmaco-economic evidence indicates that moxifloxacin is a cost effective treatment for CAP. However, data limitations and uncertainty surrounding the evolution of resistance emphasize the need for caution. As recommended by guidelines, the choice of antimicrobial should consider the local frequency of causative pathogens, the local pattern of antimicrobial resistance, and risk factors for resistant bacteria. The pharmaco-economic evidence corroborates the importance of these factors as they have an impact on the cost effectiveness of treating CAP patients with moxifloxacin. CONCLUSIONS: CAP guidelines need to take into account pharmaco-economic considerations by balancing the effectiveness of antimicrobial regimens against their costs. The pharmaco-economic value of moxifloxacin is influenced by the causative pathogens involved and resistance patterns. Therefore, it may be advisable to identify patient subgroups in which treatment with moxifloxacin is cost-effective and should be recommended by guidelines. PMID- 19678753 TI - Recognizing and treating suboptimally controlled multiple sclerosis: steps toward regaining command. AB - OBJECTIVE: The therapies available today for multiple sclerosis (MS) reduce but do not fully control disease activity. The objective of this article is to review the definitions of and treatments for suboptimally controlled MS and highlight the challenges faced by clinicians to increase awareness of recognizing and managing patients with suboptimally controlled MS. METHODS: Published literature describing treatment failure, treatment optimization paradigms or algorithms, clinical studies of therapies in patients with suboptimally controlled MS, or case reports of management of patients with suboptimally controlled MS were identified from searches of EMBASE and MEDLINE. This was supplemented with case reports and discussions from an expert panel meeting of MS specialists focused on the diagnosis and treatment of suboptimally controlled MS. RESULTS: Several groups have created recommendations for evaluating suboptimal response to disease modifying drugs (DMDs) in MS. Currently no robust evidence-based data exist to guide treatment decisions in patients who have suboptimal response to a particular therapy. In the absence of data, several treatment paradigms for suboptimally controlled MS have been proposed using a step therapy or platform therapy approach. Therapy modifications require consideration of disease- and patient-specific factors while accounting for the risk-benefit profile of the agent(s). Unapproved drugs and combination therapies should be reserved as agents of last resort because of the experimental nature of these treatments. CONCLUSIONS: In the absence of evidence-based data, identifying and treating MS patients with suboptimal response to the available platform therapies remains challenging. Developing algorithms able to quantify breakthrough disease activity and suboptimal response to DMDs in individual MS patients remains an important target for the MS community. Consideration should be given for all reasons why a particular DMD may not be working for a given patient and for the use of an individualized step therapy. PMID- 19678754 TI - Cholinesterase inhibition: is there evidence for disease-modifying effects? AB - BACKGROUND: Cholinesterase inhibitors are broadly established as first-line symptomatic therapy for Alzheimer's disease (AD). Symptomatic effects are mediated by the inhibition of acetyl- and/or butyryl-cholinesterase (AChE and/or BuChE) - the enzymes that degrade acetylcholine (ACh) in the synapse. However, ACh is also found outside the synapse ('extracellular ACh') where, among other activities, it plays a role in controlling inflammation and might impact on pathological changes. OBJECTIVE/SCOPE: New data and clinical findings are reviewed and discussed to build a preliminary case for possible disease-modifying effects of cholinesterase inhibition. FINDINGS: Trials seeking to demonstrate disease-modifying effects in subjects with mild cognitive impairment failed to reach their primary endpoints, but these failures might relate to aspects of trial methods and analyses. A re-analysis of one of these trials, using a more sensitive model controlling for factors that predict progression to AD, showed a significant delay in progression to AD with dual cholinesterase inhibition over 3 to 4 years. Taken with other evidence, it is plausible that cholinesterase inhibition might contribute to disease modification. The detection of putative disease-modifying effects may be most easily implemented in certain patient subpopulations, and genotyping studies suggest a particular role for BuChE. CONCLUSION: Long-term inhibition of BuChE might be especially important when exploring any disease-modifying effects of cholinesterase inhibitors. Elucidation of the mechanisms involved could provide insights leading to the development of new treatments that modify the development and progression of AD. PMID- 19678755 TI - Concurrent human papillomavirus-associated tonsillar carcinoma in 2 couples. AB - We describe 2 nonsmoking, nondrinking couples who developed human papillomavirus (HPV)-associated tonsillar cancer within 12 months of each other. After histopathologic evaluation, HPV L1, E2, E6, and LCR regions were amplified, and phylogenetic analysis of amplimers was determined. Quantitative polymerase chain reaction targeting HPV-16/18 L1 and E7 regions and P16 immunohistochemistry were performed. Tissues were HPV-16 positive, with distinct intercouple nucleotide differences and multiple unique intracouple similarities identified. Diffuse nuclear P16 expression was detected in tumor cells. This report demonstrates matching HPV-16 strains in 2 couples with concurrent development of tonsillar carcinoma who did not have other risk factors, revealing the potential infectious nature of oropharyngeal cancer. PMID- 19678757 TI - Determination of hepatitis C virus-infected, monocyte lineage reservoirs in individuals with or without HIV coinfection. AB - Because previous reports found an association between hepatitis C virus (HCV) coinfection and progression of human immunodeficiency virus (HIV) disease, we investigated whether HIV and HCV may reciprocally influence viral replication in monocyte lineage cells in vivo. Using a novel technique called simultaneous ultrasensitive subpopulation staining/hybridization in situ (SUSHI), we rapidly and unequivocally identified HCV reservoirs in peripheral blood from HCV-infected individuals with and without HIV coinfection. We found that HCV infects both CD14(+), CD16(+)(+) monocytic cells and CD14(+)(+), CD16(+)(+) monocytic cells but not CD14(+)(+), CD16- cells in individuals infected with HCV with or without HIV coinfection. To address these HCV tropism differences, we found that the HCV receptor CD81 is highly expressed on CD14(+), CD16(+)(+) and CD14(+)(+), CD16(+)(+) cells but not on monocytes (CD14(+)(+), CD16-). These findings have important implications for the diagnosis and treatment of HCV infection, mother to-child transmission of HCV, and possible virus-virus interactions in HCV-HIV coinfected individuals. PMID- 19678758 TI - Controlled dynamization to enhance reconstruction capacity of tissue-engineered bone in healing critically sized bone defects: an in vivo study in goats. AB - Tissue-engineered bone (TEB) has shown to be an effective alternative to conventional gold-standard autogenous bone for the repair of critically sized bone defects (CSBD). Moderate axial interfragmentary movement (IFM) has been shown to promote bone healing in conventional models. This study explored the use of IFM to enhance the capacity of TEB in the repair of CSBD using a goat model. CSBD were created in a goat model. Dynamic intramedullary rods designed to supply axial IFMs within 10% of the interfragmentary strain were used to stabilize CSBD goat femur models, whose bone defects were filled with TEB. Bone regeneration was evaluated using radionuclide bone imaging, roentgenographic analysis, periosteal callus area, computed tomography value score, biomechanical analysis, and histological observation. Compared with the static intramedullary rods, the dynamic intramedullary rod group showed an increase in early-stage callus formation and blood supply to the callus tissue, better differentiation of fibrous and cartilaginous tissue into bone tissue, improved strength and stiffness of callus tissue in late-stage healing, and overall better functional recovery of the goat femur. This showed that moderate axial IFM could promote the osteogenesis and reconstruction of TEB in vivo. PMID- 19678759 TI - Insight into the molecular pathophysiology of delayed bone healing in a sheep model. AB - Delayed and nonunions are still challenging problems. In this study, we examined the endogenous mRNA expression of genes regulating cartilage formation, bone formation, endochondral ossification, and bone remodeling during mechanically induced delayed bone healing in a large animal model. A tibial osteotomy was performed in two groups of sheep and stabilized with either a rigid external fixator leading to standard healing or with a rotationally unstable fixator leading to delayed healing. At days 4, 7, 9, 11, 14, 21, and 42 after surgery, total RNA was extracted from the callus. Gene expressions of several molecules functionally important for bone healing were studied by quantitative reverse transcriptase-polymerase chain reaction. The expression profiles were related to callus tissue composition, analyzed by histomorphometry. Histomorphometry demonstrated a delayed, prolonged chondral phase and a reduction in bone formation in the experimental group. There was no differential expression of Runx2 between both groups until day 42, but mRNA expression levels of BMP2, BMP4, BMP7, noggin, Col1a1, IGF1, TGFbeta1, OPN, MMP9, MMP13, TIMP3, TNFalpha, MCSF, RANKL, and OPG were lower in the delayed healing group at several time points. This study provides insight into the temporal periods during which various factors may be deficient during a compromised bone-healing situation. PMID- 19678760 TI - Gestational exposure to mercury leads to persistent changes in T-cell phenotype and function in adult DBF1 mice. AB - Previously, we showed that in utero exposure to mercury induced phenotypic changes in fetal immune cells. Here, we sought to determine whether the effects of in utero exposure on immune cells persisted in the adult. After overnight breeding to DBA/1 males, pregnant BALB/c dams were given either mercuric chloride in drinking water at 10 mg/L ad libitum for the duration of gestation or plain water. At the time of parturition, all dams were placed on regular drinking water. The pups (DBF(1)) were weaned and thymic and splenic tissues were harvested at 10 wk-of-age to assess T-cell phenotypes and function. Significant changes in the CD4/CD8 subsets in the thymus and spleen among mercury-exposed male and female mice were not observed. However, there was a significant reduction in splenic CD4(+)CD25(+) cells in mercury-exposed female, but not in male, mice. ConA-stimulated splenocytes from mercury-exposed mice showed significant increases in proliferative responses relative to cells from control mice, regardless of sex. Cytokine secretion was also modulated in the mercury exposed mice. In particular, the production of IL-4 and IFN by ConA-stimulated splenocytes from mercury-exposed male and female mice was significantly increased, while IL-2 and IL-10 levels were unaffected. The results of our study revealed that exposure of the developing immune system to relatively low levels of inorganic mercury could lead to persistent alterations in adult immune cell phenotypes and functions. These changes could pose a relevant health risk, if they contribute to impaired responses to pathogens and/or an increased risk for the development of atopy, asthma or possibly autoimmune diseases. PMID- 19678761 TI - Analysis of mechanisms of cell death of T-lymphocytes induced by organotin agents. AB - Organotin compounds are known to cause thymic atrophy and an accompanying deficiency of cell-mediated immunity. The study reported here focused on cell death in the thymus as a contributing factor in the induction of thymic atrophy following exposure to dibutyltin (DBTC) and tributyltin (TBTC). In an in vivo study, a reversible thymic atrophy was induced in rats by a single intraperitoneal administration (2.0 mg/kg) of DBTC or TBTC; the magnitude of this effect over a 4-d post-treatment period differed between the two agents. In in vitro studies, T-lymphocytes were isolated from thymuses of naive rats and then exposed to 1 microM DBTC or TBTC for varying periods of time. Analysis by flow cytometry showed that DBTC induced primarily necrosis, while TBTC induced apoptosis, of the cells. Activities of caspase-8, -9, and -3 were also measured; TBTC exposure caused marked increases in the activities, while DBTC exposure did not cause any significant change. TBTC exposure also appeared to induce expression of CAD (which fragments DNA), but had minimal effect on levels of the CAD inhibitor, ICAD. In contrast, DBTC exposure resulted in a larger level of ICAD expression. WST-8 and JC-1 assays were used to evaluate mitochondrial function, since a strong activation of caspase-9 by TBTC suggested mitochondrial involvement. The involvement of caspase in the activation was examined using cytochrome c expression; cytochrome expression and the loss of mitochondrial function occurred within 10 min of TBTC exposure. DBTC exposure affected the mitochondria less. These results indicated that effects on mitochondria likely played an important role in the induction of apoptosis by TBTC. The results of this study show that DBTC and TBTC induce necrosis and apoptosis of T lymphocytes, respectively, by apparently indicating different mechanisms of cell death. It follows that these increases in cell death induced by these organotin compounds likely contributed to the thymic atrophy observed in the rats here. PMID- 19678763 TI - T cell receptor (TCR) gene transfer with lentiviral vectors allows efficient redirection of tumor specificity in naive and memory T cells without prior stimulation of endogenous TCR. AB - We investigated the possibility of introducing exogenous T cell receptor (TCR) genes into T cells by lentiviral transduction, without prior stimulation of endogenous TCR with anti-CD3. TCR transfer is used to impose tumor antigen specificity on recipient T cells, but sustained activation required for retroviral transduction may affect the clinical efficacy of engineered T cells. Cytokine stimulation makes T cells susceptible to lentiviral transduction in the absence of TCR triggering, but this advantage has never been exploited for TCR transfer. Autoimmune diseases are a source of high-affinity TCRs specific for self/tumor antigens. We selected, from a patient with vitiligo, a Mart1-specific TCR based on intrinsic interchain pairing properties and functional avidity. After lentiviral transduction of human peripheral blood mononuclear cells, preferential pairing of exogenous alpha and beta chains was observed, together with effective recognition of Mart1(+) melanoma cells. We tested transduction efficiency on various T cell subsets prestimulated with interleukin (IL)-2, IL-7, IL-15, and IL-21 (alone or in combination). Both naive and unfractionated CD8(+) T cells could be transduced without requiring endogenous TCR triggering. IL-7 plus IL-15 was the most powerful combination, allowing high levels of transgene expression without inducing T cell differentiation (34 +/- 5% Mart1-TCR(+) cells in naive CD8(+) and 16 +/- 6% in unfractionated CD8(+)). Cytokine-prestimulated, Mart1-redirected naive and unfractionated CD8(+) cells expanded better than CD3 CD28-prestimulated counterparts in response to both peptide-pulsed antigen presenting cells and Mart1(+) melanoma cells. This strategy allows the generation of tumor-specific T cells encompassing truly naive T cells, endowed with an intact proliferative potential and a preserved differentiation stage. PMID- 19678762 TI - Bone grafts engineered from human adipose-derived stem cells in perfusion bioreactor culture. AB - We report engineering of half-centimeter-sized bone constructs created in vitro using human adipose-derived stem cells (hASCs), decellularized bone scaffolds, and perfusion bioreactors. The hASCs are easily accessible, can be used in an autologous fashion, are rapidly expanded in culture, and are capable of osteogenic differentiation. hASCs from four donors were characterized for their osteogenic capacity, and one representative cell population was used for tissue engineering experiments. Culture-expanded hASCs were seeded on fully decellularized native bone scaffolds (4 mm diameter x 4 mm thick), providing the necessary structural and mechanical environment for osteogenic differentiation, and cultured in bioreactors with medium perfusion. The interstitial flow velocity was set to a level necessary to maintain cell viability and function throughout the construct volume (400 microm/s), via enhanced mass transport. After 5 weeks of cultivation, the addition of osteogenic supplements (dexamethasone, sodium beta-glycerophosphate, and ascorbic acid-2-phosphate) to culture medium significantly increased the construct cellularity and the amounts of bone matrix components (collagen, bone sialoprotein, and bone osteopontin). Medium perfusion markedly improved the distribution of cells and bone matrix in engineered constructs. In summary, a combination of hASCs, decellularized bone scaffold, perfusion culture, and osteogenic supplements resulted in the formation of compact and viable bone tissue constructs. PMID- 19678764 TI - Correlations between computerized battery testing and a memory questionnaire for identification of neurocognitive impairment in HIV type 1-infected subjects on stable antiretroviral therapy. AB - Neurocognitive impairment (NCI) remains prevalent in the highly active antiretroviral therapy (HAART) era. Memory function is commonly affected. There is a need for a rapid, but sensitive screening tool. This study compares the Prospective and Retrospective Memory Questionnaire (PRMQ) and a computerised battery cognitive assessment to establish if the questionnaire has potential as a rapid screening tool for HIV-associated NCI. Neurologically asymptomatic patients with an undetectable HIV viral load on stable HAART were eligible to participate. Asymptomatic NCI (aNCI) was defined as a performance score more than 1SD below the normative mean in at least two domains of the computerised test. Memory impairment (MI) was defined as a t-score more than 1 SD below the normative mean using the PRMQ. Forty-five subjects participated. The mean age was 48 years (SD 11), the mean CD4 count was 546 cells/mul (SD 271), and 84% were male. Of subjects, 14/45 (24%) had NCI and 15/45 (33%) had MI. Two subjects had both types of impairment. No significant association was found between the presence of aNCI and MI (p = 0.229, r = 0.18, 95% CI -1.2, 0.23). aNCI was statistically significantly associated with younger age (p = 0.38, r = 0.31, 95% CI -0.02, 0.001). MI was statistically significantly associated with the set-shifting cognitive domain of the computerized battery (p = 0.04, r = 0.326) and time elapsed since HIV diagnosis (p = 0.035, r = 0.316). High rates of asymptomatic NCI were observed in this cohort, especially in younger individuals. The memory questionnaire did not reliably identify HIV-associated NCI other than executive function deficits and based on our data should therefore not be used as a rapid screening tool for this purpose. PMID- 19678765 TI - Characterization of human immunodeficiency virus type 1 populations containing CXCR4-using variants from recently infected individuals. AB - We screened 150 individuals from two recent seroconverter cohorts and found that six (4%) had CXCR4-using viruses. Clonal analysis of these six individuals, along with a seventh individual identified during clinical care as a recent seroconverter, revealed the presence of both X4- and dual-tropic variants in these recently infected adults. The ability of individual CXCR4-using variants to infect cells expressing CD4/CXCR4 or CD4/CCR5 varied dramatically. These data demonstrate that virus populations in some newly infected individuals can consist of either heterogeneous populations containing both CXCR4-using and CCR5-tropic viruses, or homogeneous populations containing only CXCR4-using viruses. The presence of CXCR4-using viruses at early stages of infection suggests that testing for viral tropism before using CCR5 antagonists may be important even in persons with known recent infection. The presence of CXCR4-using viruses in a subset of newly infected individuals could impact the efficacies of vaccine and microbicide strategies that target CCR5-tropic viruses. PMID- 19678766 TI - Comparative study of CSF neurofilaments in HTLV-1-associated myelopathy/tropical spastic paraparesis and other neurological disorders. AB - HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is a progressive CNS disease leading to corticospinal tract degeneration. Various degenerative diseases have increased neurofilament subunit concentration in cerebrospinal fluid (CSF), frequently showing hyperphosphorylation in neurofilaments. The aim of this study was to determine if there were elevated concentrations of neurofilament light subunit (NFL) and phosphorylated forms of neurofilament heavy subunit (PNFH) in HAM/TSP CSF. NF concentrations were compared with those of controls and patients with neurodegenerative diseases associated with other retroviruses (HIV-associated dementia, HAD) and a form of prion disease (familiar Creutzfeldt-Jakob, FCJD). Western blotting of CSF with antibodies against NFL showed two immunoreactive bands of 66 and 59 kDa, the latter probably corresponding to a partially degraded NFL form. The concentration of the 59-kDa form was not different in HAM/TSP compared with controls, but it was significantly increased in HAD and FCJD groups. ELISA assay for PNFH did not show differences among HAM/TSP, HAD, and control groups, while PNFH concentration was significantly elevated in FCJD. Our results show that CSF NFL and PNFH are not molecular markers of axonal damage for HAM/TSP probably due to the slow progression of this disease. NFL phosphorylation studies required previous immunoprecipitation from CSF for mass spectrometric analysis. This preliminary analysis indicated phosphorylation at S472 and at some other residues. PMID- 19678767 TI - Genetic characterization of eight full-length HIV type 1 genomes from the Democratic Republic of Congo (DRC) reveal a new subsubtype, A5, in the A radiation that predominates in the recombinant structure of CRF26_A5U. AB - In this study, we characterized HIV-1 strains from the Democratic Republic of Congo (DRC), previously described as divergent subtype A (n = 1, 97CD.KMST91) or untypable (n = 7) in the V3-V5 env region. Four strains had the same structure over the entire genome, including alternating fragments of a new subsubtype, A5, within the subtype A radiation and fragments that remain unclassified. Therefore, the cluster of new viruses represents a new circulating recombinant, CRF26_A5U. Three additional strains were unique recombinants with the newly described CRF26_A5U and subtype C. Finally, the nearly full-length sequence of 97CD.KMST91 showed that this strain also consisted of alternating fragments of a divergent subtype A lineage and unclassified fragments, although different from previously reported A and U sequences. The high genetic distances among the different CRF26 A5U strains suggest their longstanding presence in the DRC. PMID- 19678768 TI - Molecular characterization of the human immunodeficiency virus type 1 among children in Lima, Peru. AB - In Peru, there is a lack of information on molecular analysis in pediatric human immunodeficiency virus (HIV) infection. At present, the mother-to-child transmission rate is estimated at approximately 2-4%. The objective of this study was to assess the molecular epidemiology of HIV-1 in infected children. Children with suspected or confirmed pulmonary tuberculosis were evaluated at two public hospitals between 2002 and 2007. Whole blood samples were obtained from 90 HIV positive children, who were confirmed to be positive by enzyme-linked immunosorbent assay and Western blot. The specimens were subjected to envelope heteroduplex mobility assay (env HMA) followed by gag and pol gene region sequence analysis. Subtype B was found in 88 (98%) of 90 children and 2 (2%) children were subtype BF recombinants. This is the first report of recombinant HIV strains in HIV-infected children in Peru. Understanding the origin, diversity, and spread of HIV strains worldwide will be necessary for the development of an effective vaccine that targets pediatric populations throughout the world. PMID- 19678769 TI - Comparative analysis of polymorphisms in the HIV type 1 pol gene in the proviral DNA and viral RNA in the peripheral compartment. AB - The aim of this study was to evaluate the presence of mutations and polymorphisms associated with drug resistance among HIV-1-infected patients in proviral DNA and viral RNA extracted from PBMCs and plasma, respectively, in 34 HIV-1-infected patients (11 naive and 23 receiving HAART). Additional drug resistance mutations were found in only one compartment in 14 of 23 treated patients. Mutations conferring resistance to an additional drug were found in plasma in only 7 of 23 patients. A greater number of differences was found in strains in patients infected for at least more than 9 years, compared to naive patients and patients for whom the time since the first diagnosis was lower (p < 0.02). This study confirms the usefulness of simultaneous testing of different compartments for assessing drug resistance in the pol region and suggests that the heterogeneity observed in different compartments might be increased with time of infection and treatment experience. PMID- 19678770 TI - The potential utility of green tea extract as a novel treatment for cutaneous leishmaniasis. PMID- 19678771 TI - Taurine: a novel preventer of neurofibroma growth? PMID- 19678772 TI - An HMO-based prospective pilot study of energy medicine for chronic headaches: whole-person outcomes point to the need for new instrumentation. AB - OBJECTIVES: The purpose of this study was to evaluate an energy healing treatment for possible inclusion as a Kaiser Permanente Northwest (KPNW) Pain Clinic provided therapy, and to identify the appropriate number of treatment sessions for a Pain Clinic protocol, should the intervention prove successful. In addition, our intent was to document the full range of outcomes experienced by patients undergoing energy healing, including whole-person and transformative outcomes should they occur. SETTING: The setting for this study was Kaiser Permanente Northwest Pain Clinic. PARTICIPANTS: Thirteen (13) patients with chronic headache who were members of the KPNW Health Plan were recruited through flyers or mailings. METHODS: Thirteen (13) participants received at least three energy healing sessions at approximately weekly intervals. Assessments were based on pre- and post-treatment qualitative interviews. INTERVENTION: The treatment consisted of three Healing Touch sessions provided by a Certified Healing Touch Practitioner. Treatments contained elements common to all sessions, and elements that were tailored to the individual subject. RESULTS: Twelve (12) of 13 participants experienced improvement in frequency, intensity, or duration of pain after three treatments. In addition, 11 of 13 participants experienced profound shifts in their view of themselves, their lives, and their potential for healing and transformation. These changes lasted from 24 hours to more than 6 months at follow-up. CONCLUSIONS: Energy healing can be an important addition to pain management services. More in-depth qualitative research is needed to explore the diversity of outcomes facilitated by energy healing treatments. Furthermore, the development of new instrumentation is warranted to capture outcomes that reflect transformative change and changes at the level of the whole person. PMID- 19678773 TI - Combination of acupuncture and fluoxetine for depression: a randomized, double blind, sham-controlled trial. AB - BACKGROUND AND OBJECTIVE: The current pharmacological management of depression remains limited. The aim of this study was to assess the efficacy and safety of acupuncture in combination with fluoxetine as an intervention for major depressive disorder (MDD). SUBJECTS AND INTERVENTIONS: A total of 80 patients with MDD (DSM-IV) were randomized to two groups: VA group received verum acupuncture, 10 mg/day fluoxetine and placebo; SA group received sham acupuncture and 20-30 mg/day fluoxetine. Acupuncture was applied 5 times a week over a period of 6 weeks. DESIGN AND SETTINGS: A randomized, double-blind, double-dummy, sham controlled trial was conducted in Dehong Prefecture Traditional Chinese Medicine Hospital. OUTCOME MEASURES: All subjects were assessed in a double-blind fashion at four time points (i.e., baseline, the end of the second, fourth, and sixth week of treatment. The primary outcome measure was the therapeutic response rate based on the rate of total score change in the 17-item Hamilton Rating Scale for Depression (HRSD). Anxious symptoms, antidepressant side-effects, and acupuncture adverse effects were also measured additionally. RESULTS: At the end of the treatment period, the therapeutic response rates were not statistically significant between groups (80.0% for the VA group and 77.5% for the SA group, respectively). No statistical significance was found between the 2 groups in the rate of HRSD score change (z = 1.80, p = 0.07), but patients in the VA group showed better improvement than the SA group in symptoms of anxiety and side effects of antidepressant (z = 2.60, p = 0.01 and z = 23.60, p < 0.001, respectively). The overall rate of adverse events due to acupuncture was 8.75%. CONCLUSIONS: Additionally applied standardized acupuncture to low-dose fluoxetine for depression is as effective as a recommended dose of fluoxetine treatment. Depressive patients with severe anxious symptoms and/or intolerable side-effects of antidepressants can benefit from it. PMID- 19678774 TI - A randomized, placebo-controlled trial of Ginkgo biloba L. in treatment of premenstrual syndrome. AB - BACKGROUND AND OBJECTIVES: During the reproductive years, most of menstruating women experience symptoms of premenstrual syndrome (PMS), which is incapacitating in up to 10% of cases. According to complicated etiology, various therapeutic approaches have been proposed. Because PMS is a chronic situation, special attention should be paid to the side-effects of pharmacological interventions. Herbal medicine is a recent favorable therapeutic approach owing to fewer side effects. We aimed to determine the effect of Ginkgo biloba L. on the symptoms of PMS. METHODS: This was a single-blind, randomized, placebo-controlled trial conducted from November 2007 to April 2008. The students with PMS, living in dormitories of a medical university (Tehran), who met the inclusion criteria entered the study. The students filled out the daily symptom rating forms in two consecutive menstrual cycles. After we verified the PMS diagnosis in 90 students, the participants were randomly assigned to experiment and placebo groups and took G. biloba L. tablets (containing 40 mg leaf extracts) or placebo three times a day from the 16th day of the menstrual cycle to the 5th day of the next cycle. Data were collected using daily symptom rating forms. RESULTS: Eighty-five (85, 94.4%) participants completed the study. The two groups were similar in terms of demographic characteristics and baseline overall severity of symptoms. After the intervention, there was a significant decrease in the overall severity of symptoms and physical and psychologic symptoms in both Ginkgo (23.68%) and placebo (8.74%) groups (p < 0.001). However, the mean decrease in the severity of symptoms was significantly more in the Ginkgo group compared to the placebo group (p < 0.001). CONCLUSIONS: G. biloba L. can reduce the severity of PMS symptoms. Further research on active ingredients and also the efficacy and safety of various doses and treatment durations of Ginkgo are required. PMID- 19678775 TI - Vitex agnus-castus (Chaste-Tree/Berry) in the treatment of menopause-related complaints. AB - BACKGROUND: The origin of the current practice of administering Vitex agnus castus in menopause-related complaints is uncertain, but appears to be relatively recent. Here we review the evidence for this application of Vitex based on evidence from pharmacological studies and clinical research. METHODS: The mechanisms of potential relevance in the context of menopause are explored with reference to the current understanding of the endocrinology and neuroendocrinology of menopause and associated symptoms. CONCLUSIONS: We conclude that, while evidence from rigorous randomized controlled trials is lacking for the individual herb in this context, emerging pharmacological evidence supports a role for V. agnus-castus in the alleviation of menopausal symptoms and suggests that further investigation may be appropriate. PMID- 19678776 TI - The effect of guided imagery on the third stage of labor: a pilot study. AB - OBJECTIVE: The purpose of this pilot study was to test the effects of guided imagery on blood loss during the third stage of labor. DESIGN AND SETTING: This was a three-group randomized controlled trial. Recruitment was conducted from a university nurse-midwifery practice. SUBJECTS: Sixty (60) women between pregnancy weeks 36 and 38 participated. INTERVENTIONS: The women were randomized into experimental, placebo, and control groups. OUTCOME MEASURES: Intrauterine pressure catheters were used to measure uterine contraction frequency and intensity. Blood loss was measured. Analysis of variance was used to compare the three groups on continuous variables. RESULTS: Complete data were collected on 41 subjects. No significant differences were found among the groups on demographic or potentially confounding variables. CONCLUSIONS: Blood loss was nominally lower in the experimental group, but conclusions cannot be drawn due to the sample size. PMID- 19678778 TI - Evaluation of manual acupuncture at classical and nondefined points for treatment of functional dyspepsia: a randomized-controlled trial. AB - PURPOSE: Acupuncture has been used traditionally as a treatment for functional dyspepsia (FD). The goal of this trial was to examine the efficacy of acupuncture at classical points and nondefined points as a treatment for functional dyspepsia. METHODS: Sixty-eight (68) patients with functional dyspepsia, as defined by Rome-II criteria, were randomized into two groups: classical six-point acupuncture and nondefined-point acupuncture. Acupuncture was conducted three times per week for 2 weeks in a single-blind setting. To assess the effects of acupuncture, symptoms and quality of life were scored according to the Nepean Dyspepsia Index before and after acupuncture treatments. RESULTS: Acupuncture treatment significantly decreased the dyspepsia symptoms and improved the quality of life. There was no statistical difference between the acupuncture groups treated at classical and nondefined points. CONCLUSIONS: Our data show that both acupunctures at classical points and nondefined points improved the symptoms of patients with FD. However, we cannot rule out the possibilities of placebo effect in this trial. PMID- 19678777 TI - Effects of functional water on heart rate, heart rate variability, and salivary immunoglobulin a in healthy humans: a pilot study. AB - OBJECTIVE: This study was designed to explore the effects of drinking an average dietary volume of functional water on blood pressure, short-term cardiovascular variables, and salivary immunoglobulin A. DESIGN: Subjects were studied in a randomized pre-post crossover design the morning after an overnight fast. SUBJECTS: Fifteen (15) healthy nonsmoking subjects, 8 males and 7 females, aged between 15 and 49 years, participated in the study. INTERVENTION: Short-term (10 minute) electrocardiography was measured in the participants before and after drinking either control mineral or functional water. The measurements were randomized, double blinded, and held two weeks apart. Saliva was collected for 4 minutes before the start of each electrocardiograph measurement. Blood pressure was monitored at 5 minute intervals for one hour as the mean of triplicate consecutive measurements. RESULTS: Drinking 100 mL of control mineral or functional water did not alter arterial blood pressure in the 60 minutes post drinking. Drinking control mineral water led to a significant fall in the heart rate, although all time domain and power density parameters remained unaffected. Consumption of functional water resulted in a significant difference in all time domain measures and in three of the power density parameters. The heart rate fell, while RR interbeat intervals, standard deviation of the normal-to-normal heartbeats (SDNN), and the square root of the mean squared differences of successive NN intervals (RMSSD) increased. A large rise in low (LF) and high frequency (HF) power had a significant effect on total power, indicating homeodynamic balance. Elevated LF power indicated intensified spectral power at frequencies in the range of 0.1 Hz, thus improving autonomic stability. The effect on the heart rate may confirm changes in cellular metabolism. Parasympathetic pathway activation stimulated secretory immunoglobulin A on mucosal surfaces, which protects against pathogenic invasion. CONCLUSIONS: The drinking of functional water in healthy subjects elicited a rapid parasympathetic restorative response from the heart as well as stimulating mucosal humoral immunity. Functional water acts as an unconditional natural stimulus that elicits an involuntary response and facilitates physiological homeostasis. PMID- 19678779 TI - The STTEP: a Model for musculoskeletal health care in marginalized communities. AB - BACKGROUND AND OBJECTIVES: This article introduces the STTEP (Sustainable Training, Treatment, Employment Program) Model. The Model has been in operation since 1995. It provides a useful conceptual framework for policy makers, practitioners, and educators. The Model evolved from work carried out by chiropractors, myotherapists, and related health workers in poor communities through the charitable organization Hands On Health Australia. The STTEP Model grew from a recognition that poor communities mostly rely on heavy, repetitive physical labor for work. For these communities, there is little opportunity to access suitable and affordable health care requiring them to frequently live with the pain and disability associated with highly prevalent musculoskeletal conditions in their communities. The STTEP Model includes myotherapy and musculoskeletal health promotion for uncomplicated musculoskeletal conditions. CONCLUSIONS: The Model also supports training for community members and collaborates with community leaders to promote employment opportunities for graduates. The Model embraces an ethos of cultural sensitivity, corporate responsibility, and sustainability. Project Hope (Hands On Philippines Education), a program in the Philippines, is used to illustrate the Model in action. PMID- 19678780 TI - Efficacy and safety of Curcuma domestica extracts in patients with knee osteoarthritis. AB - OBJECTIVE: The objective of this study was to determine the efficacy and safety of Curcuma domestica extracts in pain reduction and functional improvement in patients with knee osteoarthritis. STUDY DESIGN AND SETTING: The design and setting were a randomized controlled study at a university hospital in Bangkok, Thailand. METHODS: One-hundred and seven (107) patients with primary knee osteoarthritis (OA) with pain score of > or =5 were randomized to receive ibuprofen 800 mg per day or C. domestica extracts 2 g per day for 6 weeks. The main outcomes were improvement in pain on level walking, pain on stairs, and functions of knee assessed by time spent during 100-m walk and going up and down a flight of stairs. The adverse events were also recorded. RESULTS: Fifty-two (52) and 55 patients were randomized to C. domestica extracts and ibuprofen groups, respectively. Baseline characteristics of the patients in both groups were not different. The mean scores of the aforementioned outcomes at weeks 0, 2, 4, and 6 were significantly improved when compared with the baseline values in both groups. There was no difference in those parameters between the patients receiving ibuprofen and C. domestica extracts, except pain on stairs (p = 0.016). No significant difference of adverse events between both groups was found (33.3% versus 44.2%, p = 0.36 in C. domestica extracts and ibuprofen groups, respectively). CONCLUSIONS: C. domestica extracts seem to be similarly efficacious and safe as ibuprofen for the treatment of knee OA. PMID- 19678781 TI - Effects of sour tea (Hibiscus sabdariffa) on lipid profile and lipoproteins in patients with type II diabetes. AB - OBJECTIVES: There is increasing evidence that intake of sour tea (Hibiscus sabdariffa) has hypoglycemic and hypolipidemic effects and may benefit patients suffering from metabolic disorders such as diabetes. The objective of the present study was to investigate the hypolipidemic effects of sour tea in patients with diabetes and compare them with those of black tea. DESIGN: In this sequential randomized controlled clinical trial, 60 patients with diabetes were recruited and randomly assigned into two groups: sour tea (ST) and black tea (BT). They were instructed to consume sour tea or black tea two times a day for 1 month. OUTCOME MEASURES: Fasting blood samples were taken at the beginning and at the end of the study for evaluation of lipids, lipoproteins, and apoproteins. RESULTS: Fifty-three (53) patients concluded the study. In the ST group, mean of high-density lipoprotein-cholesterol (HDLc) increased significantly (p = 0.002) at the end of the study, whereas changes in apolipoprotein-A1, and lipoprotein (a) were not significant. Also, a significant decrease in the mean of total cholesterol, low density lipoprotein-cholesterol, triglycerides, and Apo-B100 were seen in this group. In the BT group, only HDLc showed significant change (p = 0.002) at the end of the study and changes in the other measures were not statistically significant. CONCLUSIONS: The results of the present study showed that ST has a significant effect on blood lipid profile in patients with diabetes. PMID- 19678782 TI - Acupuncture in the treatment of diabetic bladder dysfunction. AB - OBJECTIVE: The objective of this study was to investigate the effects of acupuncture on diabetic bladder dysfunction (DBD). METHODS: This study compared 30 cases in the acupuncture group with 15 cases in the sham acupuncture group (n = 45 total). The effects of acupuncture were observed on urodynamic measurements, as well as a variety of symptoms associated with DBD. RESULTS: In the acupuncture group, five of the six urodynamic measures (maximal detrusor pressure, bladder compliance, maximal bladder capacity, bladder volume at desire to void and urge to void) demonstrated significant improvement (p < 0.05, 0.01) over the 15-day treatment period. Only one measure (bladder volume at urge to void) significantly improved (p < 0.05) in the sham acupuncture group. There were significant differences after therapy in four measures (bladder compliance, maximal bladder capacity, bladder volume at desire to void, and urge to void) between the groups (p < 0.05, 0.01). A significant difference of the changes in symptoms compared with pretreatment in the acupuncture group was observed (p < 0.05, 0.01). In 25 subjects in the acupuncture group, incontinence improved from 2.4 to 1.4. In the sham acupuncture group, incontinence deteriorated from 2.2 to 2.3. CONCLUSIONS: Our pilot study has provided evidence that acupuncture may be clinically useful for the radical treatment of DBD. PMID- 19678756 TI - Activation and coagulation biomarkers are independent predictors of the development of opportunistic disease in patients with HIV infection. AB - BACKGROUND: Activation and coagulation biomarkers were measured within the Strategies for Management of Antiretroviral Therapy (SMART) trial. Their associations with opportunistic disease (OD) in human immunodeficiency virus (HIV)-positive patients were examined. METHODS: Inflammatory (high-sensitivity C reactive protein [hsCRP], interleukin-6 [IL-6], amyloid-A, and amyloid-P) and coagulation (D-dimer and prothrombin-fragment 1+2) markers were determined. Conditional logistic regression analyses were used to assess associations between these biomarkers and risk of OD. RESULTS: The 91 patients who developed an OD were matched to 182 control subjects. Patients with an hsCRP level > or =5 microg/mL at baseline had a 3.5 higher odds of OD (95% confidence interval [CI], 1.5-8.1) than did those with an hsCRP level <1 microg/mL (P=.003, by test for trend) and patients with an IL-6 level > or =3 pg/mL at baseline had a 2.4 higher odds of OD (95% CI, 1.0-5.4) than did those with an IL-6 level <1.5 pg/mL (P=.02, by test for trend). No other baseline biomarkers predicted development of an OD. Latest follow-up hsCRP level for those with an hsCRP level > or =5 microg/mL (compared with a level <1 microg/mL; odds ratio [OR], 7.6; 95% CI, 2.0-28.5; [P=.002, by test for trend), latest amyloid-A level for those with an amyloid-A level > or =6 mg/L (compared with a level <2 mg/L; OR, 3.8; 95% CI, 1.1-13.4; P=.03, by test for trend), and latest IL-6 level for those with an IL-6 level > or =3 pg/mL (compared with a level <1.5 pg/mL; OR 2.4; 95% CI, 0.7-8.8; P=.04, by test for trend) were also associated with development of an OD. CONCLUSIONS: Higher IL-6 and hsCRP levels independently predicted development of OD. These biomarkers could provide additional prognostic information for predicting the risk of OD. PMID- 19678783 TI - A multivariate test of an expanded Andersen Health Care utilization model for complementary and alternative medicine (CAM) use in African Americans. AB - OBJECTIVES: The objectives of this study were (1) to determine which Andersen Model variables [predisposing, enabling, and need (PEN)] are related to complementary and alternative medicine (CAM) use by African Americans in the past 12 months; and (2) to determine whether the addition of disease states to the Model will explain significant variation in CAM use in the past 12 months. DESIGN: The 2002 National Health Interview Survey was used with 4256 African American adults (n = 23,828,268 weighted) selected as the study population. The dependent variable, CAM Past 12 Months, represented participants' use of at least 1 of 17 CAM modalities during the past 12 months. The Andersen Model variables [predisposing (e.g., age); enabling (e.g., insurance); and need (e.g., medical conditions)] and prevalent disease states (> or =10%) comprised the independent variables. Logistic regression analyses, incorporating the sampling weights, were employed. RESULTS: Among predisposing factors, CAM use was associated with middle aged to older, more educated, and female African Americans. Region (Northeast less likely than South) was the only significant enabling factor. Need factors had the most frequent relationships, with more medical conditions, more physician visits, better health status, prescription and over-the-counter medication use, more frequent exercise, and having activities of daily living limitations being associated with CAM use. After adjusting for PEN factors, the disease states of pain/aching joints, recurring pain, and migraine were related to CAM use. CONCLUSIONS: African American CAM users are middle-aged to older, female, educated, and have more medical conditions (especially pain-related). Users report higher utilization of "traditional" care (e.g., physician visits), indicating that CAM is likely a complement to conventional treatment in this population. Health care providers should use these factors as prompts for inquiring about CAM use in African American patients. PMID- 19678784 TI - Association between ginseng intake and mortality: Kangwha cohort study. AB - OBJECTIVE: Panax ginseng C.A. Meyer is a well-known medicinal herb in North America and Europe. The purpose of this study was to investigate the association between ginseng intake and mortality among members of the Korean population. METHODS: We followed 6282 subjects who were 55 years of age or older in March 1985 until December 31, 2003. The Cox proportional hazard regression model was used to evaluate effects of ginseng intake on mortality. RESULTS: Adjusting for age, education, occupation, drinking, smoking, self-reported chronic disease, body mass index, and blood pressure, all-cause mortality for male ginseng users was significantly lower than that for male nonusers (Hazard ratio [HR] = 0.90; 95% confidence interval [CI], 0.81-0.99). However, such an association was not observed in women (HR = 1.03; 95% CI, 0.94-1.13). Cancer-specific mortality was lower in female ginseng users than female nonusers after adjustment of relevant covariates (HR = 0.80; 95% CI, 0.60-1.08). Compared to nonusers, the HR for cancer-specific mortality in women was 0.84 in infrequent users (95% CI, 0.62 1.15) and 0.61 in frequent users (95% CI, 0.32-1.14) (p for trend, 0.09), which is not statistically significant. The cancer-specific mortality was not associated with ginseng intake in male subjects (HR = 0.95; 95% CI, 0.76-1.20). Mortality caused by cardiovascular diseases was not related to ginseng intake in both men and women. CONCLUSION: The 18.8-year progressive cohort study showed that ginseng intake decreased all-cause mortality in older males, but such life prolongation effect was not shown in women. PMID- 19678786 TI - Adult degenerative scoliosis treated by acupuncture. AB - OBJECTIVE: This report of one case illustrates the potential effect of acupuncture on low back pain and curvature progression in adult degenerative scoliosis. CLINICAL FEATURES: A 74-year-old woman experienced low back pain and kyphoscoliosis for 2 years. She received regular rehabilitation and medications for 2 years, but the curvature of the lumbar spine and backache still progressed. The Cobb angle was 31 degrees and surgical intervention was suggested to reduce the pain. INTERVENTION AND OUTCOME: Before surgery, the patient was treated with acupuncture three times per week starting on May 29, 2008. After 6 weeks of treatment, the patient's lower backache decreased and follow-up radiographs showed that the Cobb angle decreased by 10 degrees . She underwent surgery on October 3 to have a better quality of life. CONCLUSIONS: Acupuncture was associated with a reduction in the degree of curvature in this case, after 2 years of conventional medical treatment had failed to stop the backache and curvature progression. This suggests that acupuncture not only plays an important role in pain control, but can also improve curvature progression for certain patients with degenerative scoliosis. PMID- 19678785 TI - The diuretic effect in human subjects of an extract of Taraxacum officinale folium over a single day. AB - BACKGROUND: Taraxacum officinale (L.) Weber (Asteraceae) has been extensively employed as a diuretic in traditional folk medicine and in modern phytotherapy in Europe, Asia, and the Americas without prior clinical trial substantiation. OBJECTIVES: In this pilot study, a high-quality fresh leaf hydroethanolic extract of the medicinal plant T. officinale (dandelion) was ingested by volunteers to investigate whether an increased urinary frequency and volume would result. DESIGN: Volume of urinary output and fluid intake were recorded by subjects. Baseline values for urinary frequency and excretion ratio (urination volume:fluid intake) were established 2 days prior to dandelion dosing (8 mL TID) and monitored throughout a 1-day dosing period and 24 hours postdosing. RESULTS: For the entire population (n = 17) there was a significant (p < 0.05) increase in the frequency of urination in the 5-hour period after the first dose. There was also a significant (p < 0.001) increase in the excretion ratio in the 5-hour period after the second dose of extract. The third dose failed to change any of the measured parameters. CONCLUSIONS: Based on these first human data, T. officinale ethanolic extract shows promise as a diuretic in humans. Further studies are needed to establish the value of this herb for induction of diuresis in human subjects. PMID- 19678787 TI - A renal tuberculosis case: could Chinese medicine play a role? AB - CASE REPORT: A case of renal tuberculosis (TB) was treated with a multidrug therapeutic regimen (rifampicin 600 mg/day, ethambutol 800 mg/day, and isoniazid 150 mg/day), which was terminated for severe hepatotoxicity 2 months later. As an alternative therapeutic method, the patient was orally administered a Chinese herbal concoction while liver transaminases resumed normal levels. RESULTS: After 1 year of treatment, the patient recovered completely; pyuria and hematuria disappeared with negative acid-fast bacteria urine culture. The patient has been followed up for 2 years without recurrence. CONCLUSIONS: The case indicated that these Chinese herbs are useful in treating renal TB. Chinese medicine has allowed us another choice of antituberculous treatment, avoiding the hepatotoxicity of the standard therapeutic regimen. Therefore, the use of Chinese herbs has the potential of reducing the morbidity and mortality rate of this disease. PMID- 19678788 TI - Personalized medicine and complementary and alternative medicine: in search of common grounds. AB - OBJECTIVES: As complementary and alternative medicine (CAM) continues to assume influence in medical care delivery, biomedical orthodoxy has contemporaneously experienced landmark technoscientific advances, tempting analysts to question the relevance of CAM to 21st century medical provision. This article focuses on one representation of contemporary advances in biomedicine, namely, the phenomenon of personalized medicine (PM) and the technoscientific contexts for its evolution. It examines whether biomedicine's embrace of the PM concept widens the conceptual and philosophical gulf between it and CAM. DESIGN: Focusing on genomics and its translation into PM, the article finds that presently, the gene-environment dynamic is an important aspect of genomics and PM. However, there is a lopsided emphasis in the gene-environment matrix that focuses on toxicogenomics (i.e., the effect of toxins and chemicals, including drugs on genes and genetic materials). This approach to genomics ignores the role of other environmental stressors, which constitute components of an individual's health experience critical to PM. CONCLUSIONS: If this lopsided approach is addressed, in a counterintuitive way, PM has potential for engendering a confluence between biomedicine and CAM as a part of the paradox of the 21st century medical landscape. PMID- 19678791 TI - Fighting tuberculosis: old drugs, new formulations. AB - This review reports the state of the art on innovative drug delivery strategies designed for antitubercular chemotherapeutics. The introduction contains the fundamental biological background concerning tuberculosis and a review of the current antitubercular therapy, and is followed by a critical report of the micrometric and nanometric particulate systems designed and investigated to improve tuberculosis chemotherapy. PMID- 19678792 TI - Recent trends in stabilising peptides and proteins in pharmaceutical formulation considerations in the choice of excipients. AB - In the area of peptide and protein pharmaceuticals, both the physical and chemical stability of biopharmaceuticals are critical and need to be optimised when formulating a drug product, in order to optimise the outcome after processing and storage. This review focuses on the effects on the stability from various types of excipient and the choices that have to be made during formulation of drug products containing peptides or proteins. It is illustrated, through examples, how the choice of one excipient over another can affect the stability of a protein drug formulation, along with other problems linked to this choice. The excipients used in pharmaceutical preparations are limited and from an academic point of view there is a clear requirement for new excipients. PMID- 19678793 TI - Doxofylline: a promising methylxanthine derivative for the treatment of asthma and chronic obstructive pulmonary disease. AB - BACKGROUND: Doxofylline, a methylxanthine derivative, has recently drawn attention because of its better safety profile and similar efficacy over the most widely prescribed analogue, theophylline, indicated for asthma and chronic obstructive pulmonary disease. OBJECTIVE: This article attempts to discuss the pharmacodynamics/pharmacokinetics and clinical efficacy of doxofylline. METHOD: An extensive search in three electronic databases (Unbound Medline, Pubmed and Sciencedirect) and internet search engines (Scirus and Google Scholar) were used to identify the clinical studies on doxofylline. The literature search was carried out without time constraints to ensure maximum coverage of existing literature on doxofylline. RESULTS/CONCLUSION: In a relatively large number of comparative studies, doxofylline is indicated to have less affinity for alpha(1) and alpha(2) receptors than theophylline. Unlike theophylline, doxofylline does not antagonize calcium channels, nor does it interfere with the influx of calcium into the cells, which probably reduces the cardiac side effects. Moreover, it does not affect sleep rhythm, gastric secretions, heart rate and rhythm and CNS functioning. Numerous reports available regarding the better tolerability of doxofylline than theophylline prove it as a potential bronchodilator with promising pharmacological behavior. However, despite its superior safety and clinical efficacy, the potential of doxofylline has not been fully exploited. PMID- 19678794 TI - New antiretroviral drugs: a review of the efficacy, safety, pharmacokinetics, and resistance profile of tipranavir, darunavir, etravirine, rilpivirine, maraviroc, and raltegravir. AB - BACKGROUND: The introduction and approval of new antiretroviral agents in the US and Canada bring new opportunities and new challenges. Arguably, for the first time ever, clinicians have the drugs necessary to achieve the goal of suppressing HIV RNA to levels less than 50 copies/mL in even the most treatment-experienced patients and in those with extensive drug-limiting resistance mutations. However, the use of these new agents is complicated by many drug-drug interactions and--to some extent--pre-existing mutations. To derive maximum durability from the use of these newer drugs, a thorough understanding of their indications and limitations is critical. OBJECTIVE: To thoroughly review the six most recently approved or soon-to-be-approved antiretroviral drugs in the US and Canada: tipranavir, darunavir, etravirine, rilpivirine, maraviroc, and raltegravir. METHODS: Discussion of the indications for, and pharmacokinetics, resistance profile, activity, toxicity, and clinical trials results of, the six new agents. RESULTS/CONCLUSIONS: These six new agents have resulted in marked progress towards the goal of being able to provide HIV-infected individuals with the drugs necessary to achieve decades of durable suppression of HIV without substantial toxicity. PMID- 19678795 TI - Metformin: diamonds are forever. AB - Metformin has now been established as the drug of choice for the first-line management of type 2 diabetes mellitus. It reduces insulin resistance, improves glycaemic control, and can be safely combined with other classes of oral hypoglycaemic agents. Equally important, metformin has been shown to have a significant beneficial effect on cardiovascular morbidity. Moreover, this agent acts favourably on blood pressure, lipids, haemostasis and other features of the metabolic syndrome. Metformin also contributes to weight reduction and diabetes prevention. PMID- 19678796 TI - Discontinued drugs in 2008: endocrine and metabolic. AB - This perspective summarizes key compounds from the endocrine and metabolic area that were discontinued during the calendar year 2008. 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, diabetic complications, anti atherosclerosis and obesity. PMID- 19678797 TI - Is there a palliative benefit of gemcitabine plus fluoropyrimidines in patients with refractory colorectal cancer? A review of the literature previously presented: poster at the 2008 Gastrointestinal Cancer Symposium (Abstract No. 512). AB - INTRODUCTION: Fluoropyrimidines with oxaliplatin or irinotecan plus bevacizumab is the standard chemotherapy combination in patients with advanced colorectal cancer (CRC). Gemcitabine acts synergistically with fluoropyrimidines to enhance the binding of thymidylate synthase and increase inhibition of DNA synthesis. The objective of this review is to evaluate the literature for evidence of efficacy and safety of fluoropyrimidine plus gemcitabine (FG) in patients with advanced CRC. METHODS: Relevant studies were identified in PubMed, Ovid, Cochrane database and the American Society of Clinical Oncology abstracts using the following search terms: gemcitabine, fluorouracil, capecitabine and colorectal cancer. Only studies using the FG combination were selected. RESULTS: Forty-two advanced CRC patients were evaluated in two Phase I studies and the maximum tolerated dose of gemcitabine was 900 - 1,000 mg/m(2) weekly with either bolus 5-fluorouracil (5 FU) or capecitabine. A total of 216 advanced CRC patients were evaluated in six Phase II studies. Gemcitabine (750 - 1,250 mg/m2) with either 5-FU (continuous infusion or bolus) or capecitabine was administered as first-line therapy in two studies and as third-line therapy in three studies. The range reported for overall response rate was 30 - 38.3%, median time to progression was 4 - 8.3+ months and median survival was 9.8 - 18+ months. The most commonly reported grade 3 - 4 toxicities were neutropenia, thrombocytopenia and mucositis. CONCLUSIONS: Fluoropyrimidine plus gemcitabine is clinically active in patients with refractory CRC demonstrating prolonged median time to progression and acceptable toxicity only when bolus 5-FU was not used. Studies are underway to evaluate the combinations of FG with other chemotherapy or targeted agents. Meanwhile, FG may be considered for patients with advanced CRC who are refractory to primary treatment without other options or who are not eligible for clinical studies. PMID- 19678798 TI - New generation azole antifungals in clinical investigation. AB - Considerable progress in treating systemic mycoses has been achieved in the past years through development of new drugs in association with more advanced diagnostic procedures. Here, we review the pharmacological, microbiological and clinical development progress with the so-called 'second generation' triazoles: voriconazole, posaconazole, ravuconazole, isavuconazole and albaconazole. All these drugs exhibit a favourable pharmacokinetic and toxicity profile and possess high activity against resistant and emerging pathogens. However, only voriconazole and posaconazole have been adequately investigated in Phase III studies and have been approved by the regulatory agencies in the treatment and prophylaxis of invasive fungal infections, respectively. On the contrary, ravuconazole, isavuconazole and albaconazole have not been investigated in adequate clinical trials and, in the absence of proper data, the real possibilities of these agents as competitors for the treatment and prevention of invasive mycoses in the clinical setting are still unknown. The drug interactions and the variability in the absorption and/or metabolism of the triazoles, in particular voriconazole and posaconazole, may determine an unpredictable exposure of the pathogens to the antifungal treatments. Literature evidences strongly support the use of therapeutic drug monitoring for these triazoles which may be crucial for the proper management of severe invasive fungal infections. PMID- 19678799 TI - Somatostatin and somatostatin receptors: implications for neoplastic growth and cancer biology. AB - Somatostatin agonists (SM-As) are capable of achieving durable symptomatic relief and significant clinical responses in certain tumours. Herein, we review the diverse direct and indirect mechanisms of antineoplastic activity elicited by SM As as well as the hurdles that complicate their use as monotherapies in a broader range of malignancies. Emphasis is placed on recent clinical attempts to neutralise the IGF-mediated survival factor effects in the bone metastasis microenvironment in advanced prostate cancer. The first clinical trials of this 'anti-survival factor manipulation' strategy utilised the ability of SM-As to suppress the growth hormone-dependent liver-derived IGF-I bioavailability in combination with other drugs, such as dexamethasone, zolendronate and oestrogens, acting systemically and at the bone metastasis microenvironment. These regimens restored androgen ablation responsiveness in stage D3 prostate cancer patients and successfully produced objective clinical responses while only mild toxicities were observed. Furthermore, we focus on the preclinical experimental data of a targeted SM-A coupled to the super-potent doxorubicin derivative AN-201. The resulting conjugate (AN-238) has shown increased antitumour potency with a favourable toxicity profile. The potential use of novel SM-As as anticancer drugs is discussed in relation to data suggesting other direct and indirect treatment approaches pertaining to the somatostatin system. PMID- 19678800 TI - Platelet P2Y12 receptor inhibition by thienopyridines: status and future. AB - Thienopyridines have a well-established role in the treatment of coronary artery disease, especially in the setting of acute coronary syndromes and percutaneous coronary interventions. Ticlopidine, the first FDA-approved thienopyridine, was shown to be effective in reducing coronary events in high risk patients, but the original enthusiasm was hampered by concerns about its serious bone marrow toxicity. Clopidogrel a second generation thienopyridine with lesser side effects, is not only at least as effective as ticlopidine, but in combination with a low dose of aspirin, has been demonstrated to reduce the risk of major cardiovascular events in acute coronary syndrome patients in large-scale, randomised trials. Recent studies have highlighted major flaws in clopidogrel pharmacokinetics due to its delayed onset of action, and much attention has been devoted to the phenomenon of clopidogrel 'resistance'. Among the novel, third generation thienopyridines, prasugrel as compared to clopidogrel has demonstrated lower inter-patient response variability and a reduced incidence of ischaemic events, but at an increased risk of major bleeding. Currently, several studies are continuing to test new direct P2Y12 receptor antagonists, such as cangrelor and AZD6140, characterised by a faster reversal of platelet inhibition. PMID- 19678801 TI - Targeting the PI3K/AKT/mTOR signaling network in acute myelogenous leukemia. AB - BACKGROUND: The PI3K/Akt/mammalian target of rapamycin (mTOR) signaling pathway plays a central role in cell growth, proliferation and survival not only under physiological conditions but also in a variety of tumor cells. Therefore, the PI3K/Akt/mTOR axis may be a critical target for cancer therapy. OBJECTIVE: This review discusses how PI3K/Akt/mTOR signaling network is constitutively active in acute myelogenous leukemia (AML), where it strongly influences proliferation, survival and drug-resistance of leukemic cells, and how effective targeting of this pathway with pharmacological inhibitors, used alone or in combination with existing drugs, may result in suppression of leukemic cell growth, including leukemic stem cells. METHODS: We searched the literature for articles dealing with activation of this pathway in AML and highlighting the efficacy of small molecules directed against the PI3K/Akt/mTOR signaling cascade. CONCLUSIONS: The limit of acceptable toxicity for standard chemotherapy has been reached in AML. Therefore, new therapeutic strategies are needed. Targeting the PI3K/Akt/mTOR signaling network with small molecule inhibitors, alone or in combinations with other drugs, may result in less toxic and more efficacious treatment of AML patients. Efforts to exploit selective inhibitors of the PI3K/Akt/mTOR pathway that show effectiveness and safety in the clinical setting are currently underway. PMID- 19678803 TI - Corticorelin acetate injections for the treatment of peritumoral brain edema. AB - BACKGROUND: The use of corticosteroids has been shown to be effective in the management of vasogenic edema caused by brain tumors, and is currently the standard of care. The associated systemic side effects, however, can be even more debilitating than the primary disease process, ultimately warranting premature discontinuation of steroids in some patients. In response, corticorelin acetate, a synthetic targeted human corticotropin-releasing factor (hCRF) analogue, has been developed to simulate the benefits of corticosteroids in treating peritumoral brain edema (PBE), while sparing the systemic toxicity. OBJECTIVE: This article reviews the development of corticorelin acetate and its potential role in treating PBE as an alternative to standard corticosteroid therapy. METHODS: Relevant articles and abstracts were obtained from searches of the medical and chemical literature databases, as well as from the references from published articles. RESULTS/CONCLUSION: Animal studies and a Phase I randomized trial have demonstrated that hCRF is well tolerated and effective in reducing PBE and its associated signs and symptoms. In addition, the effectiveness of this drug may be helpful in sparing the use of corticosteroid therapy in patients with brain tumors, with the results from several multicenter, randomized, placebo controlled, Phase III clinical trials currently pending. PMID- 19678802 TI - Taspoglutide: a long acting human glucagon-like polypeptide-1 analogue. AB - BACKGROUND: Taspoglutide (R1583/BIM51077) is a new anti diabetic drug from Hoffmann-La Roche. The compound is to be administered as a subcutaneous injection once weekly and is also effective given bi-weekly. It is a long acting 10% formulation of (Aib 8-35) human glucagon-like polypeptide-1 (7 - 36 amides) with 93% homology with the native polypeptide. It activates the glucagon-like polypeptide-1 receptor. Phase III trials are currently in process. OBJECTIVE: To provide a critical review of taspoglutide based on available published data. METHODS: Information provided from the search on Internet has been reviewed. A clinical interpretation is given on a background of practical experience as an investigator in a clinical trial with taspoglutide. RESULTS: Search on PubMed, EMBASE and Google gave hits on six clinical studies investigating taspoglutide of which the largest accounted for > 50% of the total study population. In addition, some animal studies were identified. Significant improvement on glucose control as well as several metabolic parameters has been shown with taspoglutide. DISCUSSION: Data from the clinical trials are interpreted in a medical context. The prospects of taspoglutide in the treatment of diabetes type 2 and metabolic syndrome are discussed. CONCLUSION: Taspoglutide is a new activator of the glucagon-like polypeptide-1 receptor. It is effective when injected once weekly and less effective when injected bi-weekly. In addition to its anti diabetic properties, taspoglutide has favorable effects on body weight and significantly reduces three of five diagnostic criteria for metabolic syndrome, namely glucose, waist circumference and fasting triglyceride. PMID- 19678804 TI - Golimumab for severe asthma. AB - BACKGROUND: In severe asthma anti-TNF-alpha therapies might be effective in improving disease control based on preclinical results and on clinical short-term data. However, the long-term efficacy and safety is not known. OBJECTIVE: To discuss the data on golimumab, an anti-TNF-alpha monoclonal antibody currently used in various forms of arthritis, evaluated as an add-on anti-inflammatory therapy in severe asthma. METHODS/RESULTS: Critical appraisal of the efficacy and safety clinical data. Golimumab was not found to be generally efficacious and demonstrated an unfavourable risk-benefit ratio, but in some asthma subsets its better therapeutic effects might support its use provided the long-term safety is acceptable. CONCLUSIONS: Although at first sight the safety and efficacy data of long-term use of golimumab in severe asthma might be unsupportive, in some disease subsets it might be really effective. PMID- 19678805 TI - Metabolic pathways of trichothecenes. AB - Trichothecenes are a group of mycotoxins mainly produced by the fungi of Fusarium genus. Consumers are particularly concerned over the toxicity and food safety of trichothecenes and their metabolites from food-producing animals. The metabolism of T-2 toxin, deoxynivalenol (DON), nivalenol (NIV), fusarenon-X (FX), diacetoxyscirpenol (DAS), 3-acetyldeoxy-nivalenol (3-aDON), and 15 acetyldeoxynivalenol (15-aDON) in rodents, swine, ruminants, poultry, and humans are reviewed in this article. Metabolic pathways of these mycotoxins are very different. The major metabolic pathways of T-2 toxin in animals are hydrolysis, hydroxylation, de-epoxidation, and conjugation. After being transformed to HT-2 toxin, it undergoes further hydroxylation at C-3' to yield 3'-hydroxy-HT-2 toxin, which is considered as an activation pathway, whereas transformation from T-2 to T-2 tetraol is an inactivation pathway in animals. The typical metabolites of T-2 toxin in animals are HT-2 toxin, T-2 triol, T-2 tetraol, neosolaniol (NEO), 3' hydroxy-HT-2, and 3'-hydroxy-T-2, whereas HT-2 toxin is the main metabolite in humans. De-epoxidation is an important pathway for detoxification in animals. De epoxy products, DOM-1, and de-epoxy-NIV are the main metabolites of DON and NIV in most animals, respectively. However, the two metabolites are not found in humans. Deacetyl can occur rapidly on the acetyl derivatives, 3-aDON, 15-aDON, and FX. DAS is metabolized in animals to 15-monoacetoxyscirpenol (15-MAS) via C-4 deacetylation and then transformed to scirpentriol (SCP) via C-15 deacetylation. Finally, the epoxy is lost, yielding de-epoxy-SCP. De-epoxy-15-MAS is also the main metabolite of DAS. 15-MAS is the main metabolite in human skin. The review on the metabolism of trichothecenes will help one to well understand the fate of these toxins' future in animals and humans, as well as provide basic information for the risk assessment of them for food safety. PMID- 19678808 TI - Long-term effect of repetitive transcranial magnetic stimulation on motor function recovery after acute ischemic stroke. AB - OBJECTIVE: Although there is evidence for short term benefits of rTMS in stroke, longer term effects have not been reported. The aim of the study was to evaluate the effect of two different frequencies of rTMS on motor recovery and on cortical excitability up to 1 year post-treatment. METHODS: Forty-eight patients with acute ischemic stroke were randomly classified into three groups. The first two groups received real rTMS over motor cortex (3 and 10 Hz respectively) of the affected hemisphere and the third group received sham stimulation of the same site, daily for five consecutive days. Disability was assessed before, after fifth sessions, and then after 1, 2, 3 and 12 months. Cortical excitability was assessed for both hemispheres before and after the second and fifth sessions. RESULTS: A significant 'rTMS x time' interaction was obtained indicating that real and sham rTMS had different effects on rating scales. This was because real rTMS produced greater improvement than sham that was evident even at one year follow-up. These improvements were associated with changes in cortical excitability over the period of treatment. CONCLUSION: These results confirm that real rTMS over motor cortex can enhance and maintain recovery and may be a useful add on therapy in treatment of acute stroke patients. PMID- 19678809 TI - Increasing longevity through caloric restriction or rapamycin feeding in mammals: common mechanisms for common outcomes? AB - Significant extension of lifespan in important mammalian species is bound to attract the attention not only of the aging research community, but also the media and the wider public. Two recent papers published by Harrison et al. (2009) in Nature and by Colman et al. (2009) in Science report increased longevity of mice fed with rapamycin and of rhesus monkeys undergoing caloric restriction, respectively. These papers have generated considerable debate in the aging community. Here we assess what is new about these findings, how they fit with our knowledge of lifespan extension from other studies and what prospects this new work holds out for improvements in human longevity and human health span. PMID- 19678810 TI - Intravitreal pegaptanib sodium (Macugen) for diabetic macular oedema. AB - PURPOSE: To report the functional and anatomical outcomes resulting from the use of intravitreal pegaptanib sodium (Macugen) in patients with diabetic macular oedema (DMO). METHODS: We conducted a retrospective outcome analysis, by optical coherence tomography (OCT) and best-corrected visual acuity (BCVA), of eyes with DMO treated with intravitreal pegaptanib sodium. Moreover, we evaluated the foveal transverse photoreceptor (PR) band integrity in the OCT images at the time of the last follow-up visit. RESULTS: Sixty-three eyes of 48 patients with a minimum of 6 months of follow-up were included for analysis. Intravitreal pegaptanib was found to produce significant improvements in mean BCVA (p = 0.019) and reductions in mean central macular thickness (CMT) (p < 0.001) as soon as the 6-week follow-up. Most eyes (60/63) required a mean of 3.03 +/- 0.9 repeated treatments, over a mean follow-up period of 6.7 +/- 1.2 months, to achieve significant improvements in mean BCVA (p < 0.001) and mean CMT (p < 0.001). In our series, the lower visual acuities tended to congregate in the group with the less-defined PR band (p < 0.001) and the lower CMT tended to congregate in the group with the best-defined PR band (p = 0.04), even though the higher CMT did not tend to congregate in the group with the less-defined PR band. CONCLUSION: Our findings demonstrate that selective inhibition by intravitreal pegaptanib sodium of vascular endothelial growth factor (VEGF)-165 may produce a clinically meaningful and statistically significant benefit in the treatment of DMO. PMID- 19678811 TI - Significant improvements in near vision, reading speed, central visual field and related quality of life after ranibizumab treatment of wet age-related macular degeneration. AB - PURPOSE: To investigate the effects on near visual acuity, reading speed, central visual field and related quality of life of ranibizumab treatment of wet age related macular degeneration (AMD). METHODS: The study was a prospective, non comparative consecutive case series, followed for 3 months and investigator driven. Thirty eyes of 30 patients with wet AMD were included, mean age 75 years (range 69-95 years). In addition to a full ophthalmological examination- including best-corrected visual acuity (BCVA; Early Treatment Diabetic Research Study chart), fundus biomicroscopy, fundus photography, fluorescein angiography, indocyanine green angiography (occult cases) and ocular coherence tomography- near visual acuity, reading speed, central visual field and quality of life for related activities were also investigated at baseline and at 3 months after ranibizumab treatment. RESULTS: Mean BCVA increased from 62 +/- 11 to 66 +/- 14 letters at 3 months (7%; p = 0.018). Near vision improved from 9 +/- 5 to 6 +/- 3 points (33%; p = 0.0006) and reading speed increased from 59 +/- 40 to 85 +/- 50 words/min (44%; p < 0.0001). The mean deviation from normal of the visual field improved from -9 +/- 7 to -6 +/- 5 dB (33%; p < 0.0001). Quality of life improved for distance activities from 54 +/- 28 to 63 +/- 28 points (17%; p < 0.0001) but significantly (p = 0.024) more for near activities, from 49 +/- 26 to 63 +/- 26 points (29%; p < 0.0001). Reading newspaper text in the group in which the better eye was treated showed the highest increase in quality of life score of all: 116%. CONCLUSION: The increase in BCVA after ranibizumab treatment is well established. The present study also showed significant improvements in other important visual qualities, such as near visual acuity, reading speed, central visual field and several activities influencing quality of life. The improvement was greater for near activities than for distance activities. Therefore, the beneficial effects of ranibizumab treatment shown here are more extensive than those reported previously. PMID- 19678812 TI - Effects of bisacodyl on ascending colon emptying and overall colonic transit in healthy volunteers. AB - BACKGROUND: The mechanism of action of bisacodyl in the unprepared human colon remains unclear. AIM: To evaluate the effect of oral bisacodyl on the overall and regional colonic transit in humans. METHODS: In a double-blind, randomized, placebo-controlled study of 25 healthy participants, effects of oral bisacodyl (5 mg p.o. per day) and placebo on colonic transit were compared. A validated scintigraphy method was used to measure colonic transit. The primary transit endpoints, ascending colon emptying t(1/2) and geometric centre of colon isotope at 24 h (overall transit), were compared (Wilcoxon rank sum test). RESULTS: There were significant treatment effects on ascending colon t(1/2), with the bisacodyl group demonstrating accelerated emptying [median 6.5 h, interquartile range 5.0 8.0 h] relative to the placebo group [11.0 h (7.0-17.1); P = 0.03]. Numerical differences in colonic geometric centre 24 h [bisacodyl median 3.0 (2.2-3.8), placebo 4.0 (3.1-4.6)] were not significant (P = 0.19). There were no significant differences observed in geometric centre 4 h. CONCLUSION: Oral 5 mg bisacodyl accelerates ascending colon in the unprepared colon in healthy adults; this action may contribute to the drug's efficacy in constipation. PMID- 19678813 TI - Clinical trial: ulcerative colitis maintenance treatment with 5-ASA: a 1-year, randomized multicentre study comparing MMX with Asacol. AB - BACKGROUND: 5-ASA-MMX (1.2 g/tablet) is a 5-aminosalicylic acid formulation, designed for once-daily dosing in the treatment of ulcerative colitis. AIM: To evaluate the efficacy and safety of 5-ASA-MMX (2.4 g/day, once daily), compared with Asacol (2.4 g/day, twice daily) in the maintenance of left-sided UC, through a double-blind, double-dummy, parallel-group, randomized, comparator study. METHODS: In all, 331 patients with UC were randomized to receive either 5-ASA-MMX 2.4 g/day, once daily, or Asacol 2.4 g/day, twice daily, for 12 months. All patients were in remission for >or=1 month prior to the trial, with >or=1 documented relapse in the previous year. The co-primary endpoints of this study were the proportion of patients in clinical, and clinical and endoscopic remission following 12 months' treatment. RESULTS: In the intent-to-treat population, excluding those with major protocol deviations, 68.0 and 65.9% patients in the 5-ASA-MMX and Asacol groups, respectively, were in clinical remission (P = 0.69), and 60.9 and 61.7% of patients, respectively, were in clinical and endoscopic remission (P = 0.89). Diary card data revealed statistically significant treatment differences favouring 5-ASA-MMX. Both treatments were similarly tolerated. CONCLUSIONS: Once-daily 5-ASA-MMX is similarly effective with a comparable safety profile to Asacol administered twice daily, for the maintenance treatment of ulcerative colitis. PMID- 19678814 TI - Review article: the modern management of portal vein thrombosis. AB - BACKGROUND: Portal vein thrombosis (PVT) is an important cause of portal hypertension. It may occur as such with or without associated cirrhosis and hepatocellular carcinoma. Information on its management is scanty. AIM: To provide an update on the modern management of portal vein thrombosis. Information on portal vein thrombosis in patients with and without cirrhosis and hepatocellular carcinoma is also updated. METHODS: A pubmed search was performed to identify the literature using search items portal vein thrombosis-aetiology and treatment and portal vein thrombosis in cirrhosis and hepatocellular carcinoma. RESULTS: Portal vein thrombosis occurs because of local inflammatory conditions in the abdomen and prothrombotic factors. Acute portal vein thrombosis is usually symptomatic when associated with cirrhosis and/or superior mesenteric vein thrombosis. Anticoagulation should be given for 3-6 months if detected early. If prothrombotic factors are identified, anticoagulation should be given lifelong. Chronic portal vein thrombosis usually presents with well tolerated upper gastrointestinal bleed. It is diagnosed by imaging, which demonstrates a portal cavernoma in place of a portal vein. Anticoagulation does not have a definite role, but bleeds can be treated with endotherapy or shunt surgery. Rarely liver transplantation may be considered. CONCLUSION: Role of anticoagulation in chronic portal vein thrombosis needs to be further studied. PMID- 19678815 TI - DNA content in the diagnostic biopsy for benign-adjacent and cancer-tissue areas predicts the need for treatment in men with T1c prostate cancer undergoing surveillance in an expectant management programme. AB - STUDY TYPE: Prognosis (case series). LEVEL OF EVIDENCE: 4. OBJECTIVE: To assess the DNA content in benign-adjacent and cancer-tissue areas of a diagnostic biopsy, to predict which patients would subsequently develop an unfavourable biopsy necessitating treatment for prostate cancer in the expectant management (EM) programme. PATIENTS AND METHODS: Of 71 patients who had benign-adjacent and cancer-tissue areas of diagnostic biopsies available, 39 developed unfavourable biopsies (Gleason score > or =7, Gleason pattern 4/5, three or more cores positive for cancer, >50% of any core involved with cancer), while 32 maintained favourable biopsies on annual surveillance examination (median follow-up 3.7 years). DNA content was measured on Feulgen-stained biopsy sections using an automatic imaging system (AutoCyte(TM), TriPath Imaging Inc, Burlington, NC, USA). Cox proportional-hazard regression and Kaplan-Meier plots were used to identify significant predictors for unfavourable biopsy conversion. RESULTS: Univariately, DNA content measurements i.e. an excess of optical density (OD) in the benign-adjacent tissuer area, and the sd of the OD in the cancer tissue were significant, with a hazard ratio and 95% confidence interval of 2.58 (1.17-5.68; P = 0.019) and 5.36 (1.89-15.24; P = 0.002), respectively, for predicting unfavourable biopsy conversion that required intervention. Also, several other DNA content measurements in benign-adjacent and cancer-tissue areas showed a trend to statistical significance. Further, benign-adjacent excess of OD (3.12, 1.4-6.95; P = 0.005) and cancer sd of OD (5.88, 2.06-16.82; P = 0.001) remained significant in the multivariate model to predict unfavourable biopsy conversion. Patients with benign-adjacent excess of OD > 25.0 and cancer sd of OD of >4.0 had the highest risk for unfavourable biopsy conversion (P < 0.001). CONCLUSIONS: DNA content measurements in the benign-adjacent and cancer-tissue areas appear to be useful for predicting unfavourable biopsy conversion (a recommendation for intervention) on annual surveillance examinations in the EM programme. PMID- 19678816 TI - Evidence-based urology in practice: likelihood ratios. PMID- 19678817 TI - Stem cells in regenerative urology of the bladder. PMID- 19678818 TI - Evidence for an 'anti-ageing' product may not be so clear as it appears. PMID- 19678820 TI - Genetic analysis of desmoglein 3 (DSG3) sequence variants in patients with pemphigus vulgaris. PMID- 19678821 TI - Peripheral regional aseptic neutrophilic lymphadenopathy associated with pyoderma gangrenosum. PMID- 19678822 TI - Merkel cell polyomavirus DNA detection in lesional and nonlesional skin from patients with Merkel cell carcinoma or other skin diseases. AB - Background A novel polyomavirus, the Merkel cell polyomavirus (MCPyV), has recently been identified in Merkel cell carcinoma (MCC). Objectives To investigate the specificity of this association through the detection, quantification and analysis of MCPyV DNA in lesional and nonlesional skin biopsies from patients with MCC or with other cutaneous diseases, as well as in normal skin from clinically healthy individuals. Methods DNA was extracted from lesional and nonlesional skin samples of patients with MCC or with other cutaneous diseases and from normal-appearing skin of clinically healthy subjects. MCPyV DNA was detected by polymerase chain reaction (PCR) and quantified by real time PCR. Additionally, the T antigen coding region was sequenced in eight samples from seven patients. Results MCPyV DNA was detected in 14 of 18 (78%) patients with MCC, five of 18 (28%) patients with other skin diseases (P = 0.007) and one of six (17%) clinically healthy subjects. In patients with MCC, viral DNA was detected in nine of 11 (82%) tumours and in 10 of 14 (71%) nontumoral skin samples (P = 0.66). MCPyV DNA levels were higher in MCC tumours than in nontumoral skin from patients with MCC, and than in lesional or nonlesional skin from patients with other cutaneous disorders. Signature mutations in the T antigen gene were not identified in the two MCC tumour specimens analysed. Conclusions High prevalence and higher levels of MCPyV DNA in MCC supports a role for MCPyV in tumorigenesis. However, the high prevalence of MCPyV in the nontumoral skin and in subjects without MCC suggests that MCPyV is a ubiquitous virus. PMID- 19678823 TI - Therapeutic efficacy of Bifidobacterium longum-mediated human granulocyte colony stimulating factor and/or endostatin combined with cyclophosphamide in mouse transplanted tumors. AB - Granulocyte colony-stimulating factor (GCSF) is frequently used as an adjunctive agent in tumor chemotherapy. Bifidobacterium longums (B. longum) attracted researchers' interests due to its enhancement of immunity and selective location in solid tumors. B. longum-pBV22210-endostatin (Endo) was proved to have a definite inhibitive effect on tumor growth in our previous study. In the present study, we evaluated the effects of B. longum-pBV22210-GCSF and/or B. longum pBV22210-Endo combined with cyclophosphamide (CTX) on H22 and S180 tumor-bearing mice. Based on our previous work, the plasmid pBV22210-GCSF was constructed and transformed by electroporation into B. longum. The B. longum-pBV22210-GCSF and/or B. longum-pBV22210-Endo combined with CTX were applied to treat H22 and S180 tumor-bearing mice. A leukocyte count was carried out and the tumor inhibition rate was calculated after treatment. In our study, CTX combined with B. longum pBV22210-GCSF significantly raised the leukocyte level of tumor-bearing mice, while combined with B. longum-pBV22210-GCSF alone or B. longum-pBV22210-Endo alone combinations with CTX inhibited tumor growth by over 65%. The results showed that B. longum-pBV22210-GCSF had an effective antagonistic effect on bone marrow inhibited by CTX and could inhibit tumor growth when it was combined with B. longum-pBV22210-Endo and CTX. Our results provide an enhanced understanding of B. longum and GCSF as well as their potential as an adjunctive approach in cancer gene therapy. PMID- 19678824 TI - Localization of pathology in asthma. PMID- 19678825 TI - Pedunculated cutaneous spindle cell/pleomorphic lipoma. AB - Spindle cell/pleomorphic lipoma is an infrequently seen benign adipose tissue tumor. This tumor, mostly arising from the subcutaneous tissue, usually affects male patients and occurs in back, shoulders, head and neck area. It is rarely localized to dermis. Cutaneous spindle cell/pleomorphic lipoma differs from its subcutaneous counterpart by wider anatomical distribution, female predilection and infiltrative margins. We here present a pedunculated type, cutaneous pleomorphic lipoma case localized to the nasolabial region of 59-year-old man. PMID- 19678826 TI - Fibro-osseous pseudotumor of the digit: a clinicopathological study of 17 cases. AB - Fibro-osseous pseudotumor (FP) of the digit is a rare, non-neoplastic heterotopic ossifying lesion involving the subcutaneous tissues of the digits. To date, there are only a few published series in the literature. Our study of 17 cases, retrieved from the authors' referral archives, shows that the condition chiefly affects young to middle-aged adults (median = 34 years), with a slight female predominance and involves the fingers (n = 8) and toes (n = 8). One identical lesion was identified on the forehead (n = 1). Lesional size ranges from 0.8 to 5.6 cm. Treatment was by surgical excision. Histologically, 14 cases show a fairly well-circumscribed dermal (n = 10) or dermal and subcutaneous (n = 7) lesion with surface ulceration (n = 7). The lesion is composed of fascicles of variably cellular, spindle-shaped cells [calponin (n = 14) and smooth muscle actin (SMA) (n = 11) positivity], with minimal to mild atypia (n = 5), dispersed in a myxoid stroma, focally reminiscent of nodular fasciitis. At least focal irregular trabeculae with osteoid formation and osteoblastic rimming are seen in all cases. The main differential diagnosis is an extraskeletal osteosarcoma; however, this afflicts an older age group, with prominent cytological atypia and atypical mitoses. Clinical follow-up (range: 18 months - 14 years, n = 12) reveals evidence of local recurrence in some cases (n = 2), but no evidence of metastases. In conclusion, we report an additional 17 cases of this rare lesion to increase awareness amongst dermatopathologists. PMID- 19678828 TI - Comparative analysis of the widespread and conserved PB1-like viruses infecting Pseudomonas aeruginosa. AB - We examined the genetic diversity of lytic Pseudomonas aeruginosa bacteriophage PB1 and four closely related phages (LBL3, LMA2, 14-1 and SN) isolated throughout Europe. They all encapsulate linear, non-permuted genomes between 64 427 and 66 530 bp within a solid, acid-resistant isometric capsid (diameter: 74 nm) and carry non-flexible, contractile tails of approximately 140 nm. The genomes are organized into at least seven transcriptional blocks, alternating on both strands, and encode between 88 (LBL3) and 95 (LMA2) proteins. Their virion particles are composed of at least 22 different proteins, which were identified using mass spectrometry. Post-translational modifications were suggested for two proteins, and a frameshift hotspot was identified within ORF42, encoding a structural protein. Despite large temporal and spatial separations between phage isolations, very high sequence similarity and limited horizontal gene transfer were found between the individual viruses. These PB1-like viruses constitute a new genus of environmentally very widespread phages within the Myoviridae. PMID- 19678829 TI - Biofilms: the environmental playground of Legionella pneumophila. AB - Legionella pneumophila, the aetiological agent of 90% of legionellosis cases, is a common inhabitant of natural and anthropogenic freshwater environments, where it resides in biofilms. Biofilms are defined as complex, natural assemblages of microorganisms that involve a multitude of trophic interactions. A thorough knowledge and understanding of Legionella ecology in relation to biofilm communities is of primary importance in the search for innovative and effective control strategies to prevent the occurrence of disease cases. This review provides a critical update on the state-of-the-art progress in understanding the mechanisms and factors affecting the biofilm life cycle of L. pneumophila. Particular emphasis is given to discussing the different strategies this human pathogen uses to grow and retain itself in biofilm communities. Biofilms develop not only at solid-water interfaces (substrate-associated biofilms), but also at the water-air interface (floating biofilms). Disturbance of the water surface can lead to liberation of aerosols derived from the floating biofilm into the atmosphere that allow transmission of biofilm-associated pathogens over considerable distances. Recent data concerning the occurrence and replication of L. pneumophila in floating biofilms are also elaborated and discussed. PMID- 19678830 TI - Comparative analyses of actinobacterial genomic fragments from Lake Kinneret. AB - The high genomic G+C group of Actinobacteria possesses a variety of physiological and metabolic properties, and exhibits diverse lifestyles and ecological distribution. In recent years, Actinobacteria have been found to frequently dominate samples obtained from freshwater samples. Furthermore, phylogenetic analyses have shown that 16S rRNA genes from uncultured actinobacterial freshwater samples cluster in four distinct lineages. While these lineages are abundant, little is known about them and currently no pure-culture representatives or genomic fragments of them are available. In a screen of a genomic library from the moderately eutrophic freshwater Lake Kinneret, five fosmid clones containing actinobacterial genomic fragments were found. Three approximately 40 kb genomic fragments were chosen for sequencing. Fosmids K003 and K005 showed high similarity and were affiliated with the acIV actinobacterial freshwater lineage. Fosmid K004 was affiliated with the highly abundant acI lineage. A comparative genomic analysis revealed high synteny between the two freshwater clones K003 and K005 but a lower synteny between these two and the K004 fosmid. Fosmids K003 and K005 share an identical arrangement of arginine biosynthesis gene while K004 showed a slightly different arrangement by lacking the argF gene. Fosmid Ant4E12, an Antarctic actinobacterial clone, showed a higher synteny with K003/5 than K004 and a similar arginine operon, but in a different genomic context. The Clusters of Orthologous Groups categories assignment of the three fosmids yielded genes that were mostly involved in amino acid and nucleotide metabolism, as well as transport and ribosomal RNA translation, structure and biogenesis. These genomic fragments represent the first sequences to be published from these lineages, providing a cornerstone for future work on this environmentally dominant group. PMID- 19678831 TI - Eukaryotic diversity and phylogeny using small- and large-subunit ribosomal RNA genes from environmental samples. AB - The recent introduction of molecular techniques in eukaryotic microbial diversity studies, in particular those based in the amplification and sequencing of small subunit ribosomal DNA (SSU rDNA), has revealed the existence of an unexpected variety of new phylotypes. The taxonomic ascription of the organisms bearing those sequences is generally deduced from phylogenetic analysis. Unfortunately, the SSU rDNA sequence alone has often not enough phylogenetic information to resolve the phylogeny of fast-evolving or very divergent sequences, leading to their misclassification. To address this problem, we tried to increase the phylogenetic signal by amplifying the complete eukaryotic rDNA cluster [i.e. the SSU rDNA, the internal transcribed spacers, the 5.8S rDNA and the large-subunit (LSU) rDNA] from environmental samples, and sequencing the SSU and LSU rDNA part of the clones. Using marine planktonic samples, we showed that surveys based on either SSU or SSU + LSU rDNA retrieved comparable diversity patterns. In addition, phylogenetic trees based on the concatenated SSU + LSU rDNA sequences showed better resolution, yielding good support for major eukaryotic groups such as the Opisthokonta, Rhizaria and Excavata. Finally, highly divergent SSU rDNA sequences, whose phylogenetic position was impossible to determine with the SSU rDNA data alone, could be placed correctly with the SSU + LSU rDNA approach. These results suggest that this method can be useful, in particular for the analysis of eukaryotic microbial communities rich in phylotypes of difficult phylogenetic ascription. PMID- 19678833 TI - Acute stroke management: avoiding procrastination, the best way to optimize care delivery. PMID- 19678832 TI - Stratified bacterial community in the bladder of the medicinal leech, Hirudo verbana. AB - Most animals harbour symbiotic microorganisms inside their body, where intimate interactions occur between the partners. The medicinal leech, Hirudo verbana, possesses 17 pairs of excretory bladders that harbour a large number of intracellular and extracellular symbiotic bacteria. In this study, we characterized the bladder symbionts using molecular phylogenetic analyses, transmission electron microscopy (TEM) and fluorescence in situ hybridization (FISH). Restriction fragment length polymorphism (RFLP) and sequence analyses of 16S rRNA gene clone libraries suggested that six bacterial species co-colonize the leech bladders. Phylogenetic analyses revealed that these species belong to the alpha-Proteobacteria (Ochrobactrum symbiont), beta-Proteobacteria (Beta-1 and Beta-2 symbionts), delta-Proteobacteria (Bdellovibrio symbiont) and Bacteroidetes (Niabella and Sphingobacterium symbionts). Species-specific PCR detection and FISH confirmed the localization of the symbiotic bacteria in the bladders. The Ochrobactrum, Beta-1, Bdellovibrio and Sphingobacterium symbionts were consistently detected in 13 leeches from two populations, while infection rate of the other symbionts ranged between 20% and 100% in the two leech populations. Transmission electron microscopy observations of the bladders revealed epithelial cells harbouring a number of intracellular bacilli and an additional type of extracellular, rod-shaped bacteria in the luminal region. Fluorescence in situ hybridization with group-specific oligonucleotide probes revealed the spatial organization of the bacterial species in the bladder: the Ochrobactrum symbiont was located intracellularly inside epithelial cells; the Bacteroidetes were localized close to the epithelium in the lumen of the bladder; and the Bacteroidetes layer was covered with dense beta-proteobacterial cells. These results clearly demonstrate that a simple but organized microbial community exists in the bladder of the medicinal leech. PMID- 19678835 TI - Pre-orgasmic headache and cervical carotid artery dissection. PMID- 19678834 TI - Fava beans and Parkinson's disease: useful 'natural supplement' or useless risk? PMID- 19678836 TI - Macrocypins, a family of cysteine protease inhibitors from the basidiomycete Macrolepiota procera. AB - A new family of cysteine protease inhibitors from the basidiomycete Macrolepiota procera has been identified and the family members have been termed macrocypins. These macrocypins are encoded by a family of genes that is divided into five groups with more than 90% within-group sequence identity and 75-86% between-group sequence identity. Several differences in the promoter and noncoding sequences suggest regulation of macrocypin expression at different levels. High yields of three different recombinant macrocypins were produced by bacterial expression. The sequence diversity was shown to affect the inhibitory activity of macrocypins, the heterologously expressed macrocypins belonging to different groups showing differences in their inhibitory profiles. Macrocypins are effective inhibitors of papain and cysteine cathepsin endopeptidases, and also inhibit cathepsins B and H, which exhibit both exopeptidase and endopeptidase activities. The cysteine protease legumain is inhibited by macrocypins with the exception of one representative that exhibits, instead, a weak inhibition of serine protease trypsin. Macrocypins exhibit similar basic biochemical characteristics, stability against high temperature and extremes of pH, and inhibitory profiles similar to those of clitocypin from Clitocybe nebularis, the sole representative of the I48 protease inhibitor family in the merops database. This suggests that they belong to the same merops family of cysteine protease inhibitors, the mycocypins, and substantiates the establishment of the I48 protease inhibitor family. PMID- 19678837 TI - Weak oligomerization of low-molecular-weight protein tyrosine phosphatase is conserved from mammals to bacteria. AB - The well-characterized self-association of a mammalian low-molecular-weight protein tyrosine phosphatase (lmwPTP) produces inactive oligomers that are in equilibrium with active monomers. A role of the inactive oligomers as supramolecular proenzymes has been suggested. The oligomerization equilibrium of YwlE, a lmwPTP from Bacillus subtilis, was studied by NMR. Chemical shift data and NMR relaxation confirm that dimerization takes place through the enzyme's active site, and is fully equivalent to the dimerization previously characterized in a eukaryotic low-molecular-weight phosphatase, with similarly large dissociation constants. The similarity between the oligomerization of prokaryotic and eukaryotic phosphatases extends beyond the dimer and involves higher order oligomers detected by NMR relaxation analysis at high protein concentrations. The conservation across different kingdoms of life suggests a physiological role for lmwPTP oligomerization in spite of the weak association observed in vitro. Structural data suggest that substrate modulation of the oligomerization equilibrium could be a regulatory mechanism leading to the generation of signaling pulses. The presence of a phenylalanine residue in the dimerization site of YwlE, replacing a tyrosine residue conserved in all eukaryotic lmwPTPs, demonstrates that lmwPTP regulation by oligomerization can be independent from tyrosine phosphorylation. PMID- 19678839 TI - Atomic-resolution structure of reduced cyanobacterial cytochrome c6 with an unusual sequence insertion. AB - The structure of the reduced form of cytochrome c(6) from the mesophilic cyanobacterium Synechococcus sp. PCC 7002 has been determined at 1.2 A and refined to an R-factor of 0.107. This protein is unique among all known cytochromes c(6), owing to the presence of an unusual seven-residue insertion, KDGSKSL(44-50), which differs from the insertion found in the recently discovered plant cytochromes c(6A). Furthermore, the present protein is unusual because of its very high content (36%) of the smallest residues (glycine and alanine). The structure reveals that the overall fold of the protein is similar to that of other class I c-type cytochromes, despite the presence of the specific insertion. The insertion is located within the most variable region of the cytochrome c(6) sequence, i.e. between helices II and III. The first six residues [KDGSKS(44-49)] form a loop, whereas the last residue, Leu50, extends the N-terminal beginning of helix III. Several specific noncovalent interactions are found inside the insertion, as well as between the insertion and the rest of the protein. The crystal structure contains three copies of the cytochrome c(6) molecule per asymmetric unit, and is characterized by an unusually high packing density, with solvent occupying barely 17.58% of the crystal volume. PMID- 19678838 TI - Structural recognition of an optimized substrate for the ephrin family of receptor tyrosine kinases. AB - Ephrin receptor tyrosine kinase A3 (EphA3, EC 2.7.10.1) is a member of a unique branch of the kinome in which downstream signaling occurs in both ligand- and receptor-expressing cells. Consequently, the ephrins and ephrin receptor tyrosine kinases often mediate processes involving cell-cell contact, including cellular adhesion or repulsion, developmental remodeling and neuronal mapping. The receptor is also frequently overexpressed in invasive cancers, including breast, small-cell lung and gastrointestinal cancers. However, little is known about direct substrates of EphA3 kinase and no chemical probes are available. Using a library approach, we found a short peptide sequence that is a good substrate for EphA3 and is suitable for co-crystallization studies. Complex structures show multiple contacts between kinase and substrates; in particular, two residues undergo conformational changes and by mutation are found to be important for substrate binding and turnover. In addition, a difference in catalytic efficiency between EPH kinase family members is observed. These results provide insight into the mechanism of substrate binding to these developmentally integral enzymes. PMID- 19678840 TI - Structural insight into the evolutionary and pharmacologic homology of glutamate carboxypeptidases II and III. AB - Glutamate carboxypeptidase III (GCPIII) is a metalloenzyme that belongs to the transferrin receptor/glutamate carboxypeptidase II (GCPII; EC 3.4.17.21) superfamily. GCPIII has been studied mainly because of its evolutionary relationship to GCPII, an enzyme involved in a variety of neuropathologies and malignancies, such as glutamatergic neurotoxicity and prostate cancer. Given the potential functional and pharmacological overlap between GCPIII and GCPII, studies addressing the structural and physiological properties of GCPIII are crucial for obtaining a deeper understanding of the GCPII/GCPIII system. In the present study, we report high-resolution crystal structures of the human GCPIII ectodomain in a 'pseudo-unliganded' state and in a complex with: (a) L-glutamate (a product of hydrolysis); (b) a phosphapeptide transition state mimetic, namely (2S,3'S)-{[(3'-amino-3'-carboxy-propyl)-hydroxyphosphinoyl]methyl}-pentanedioic acid; and (c) quisqualic acid, a glutamate biostere. Our data reveal the overall fold and quaternary arrangement of the GCPIII molecule, define the architecture of the GCPIII substrate-binding cavity, and offer an experimental evidence for the presence of Zn(2+) ions in the bimetallic active site. Furthermore, the structures allow us to detail interactions between the enzyme and its ligands and to characterize the functional flexibility of GCPIII, which is essential for substrate recognition. A comparison of these GCPIII structures with the equivalent GCPII complexes reveals differences in the organization of specificity pockets, in surface charge distribution, and in the occupancy of the co-catalytic zinc sites. The data presented here provide information that should prove to be essential for the structurally-aided design of GCPIII-specific inhibitors and might comprise guidelines for future comparative GCPII/GCPIII studies. PMID- 19678841 TI - Dissecting the role of the N-terminal metal-binding domains in activating the yeast copper ATPase in vivo. AB - In yeast, copper delivery to the trans-Golgi network involves interactions between the metallo-chaperone Atx1 and the N-terminus of Ccc2, the P-type ATPase responsible for copper transport across trans-Golgi network membranes. Disruption of the Atx1-Ccc2 route leads to cell growth arrest in a copper-and-iron-limited medium, a phenotype allowing complementation studies. Coexpression of Atx1 and Ccc2 mutants in an atx1Delta ccc2Delta strain allowed us to study in vivo Atx1 Ccc2 and intra-Ccc2 domain-domain interactions, leading to active copper transfer into the trans-Golgi network. The Ccc2 N-terminus encloses two copper-binding domains, M1 and M2. We show that in vivo Atx1-M1 or Atx1-M2 interactions activate Ccc2. M1 or M2, expressed in place of the metallo-chaperone Atx1, were not as efficient as Atx1 in delivering copper to the Ccc2 N-terminus. However, when the Ccc2 N-terminus was truncated, these independent metal-binding domains behaved like functional metallo-chaperones in delivering copper to another copper-binding site in Ccc2 whose identity is still unknown. Therefore, we provide evidence of a dual role for the Ccc2 N-terminus, namely to receive copper from Atx1 and to convey copper to another domain of Ccc2, thereby activating the ATPase. At variance with their prokaryotic homologues, Atx1 did not activate the Ccc2 derived ATPase lacking its N-terminus. PMID- 19678842 TI - Dissection of LolB function--lipoprotein binding, membrane targeting and incorporation of lipoproteins into lipid bilayers. AB - Escherichia coli cells express at least 90 species of lipoprotein. LolB is one of the essential outer membrane lipoproteins, being involved in the last step of lipoprotein sorting. It accepts lipoproteins from a periplasmic molecular chaperone, LolA, and mediates the outer membrane anchoring of lipoproteins through a largely unknown mechanism. It has been shown previously that a LolB derivative, mLolB, lacking an N-terminal acyl chain, can bind lipoproteins. We examined how the lack of an N-terminal anchor affects the outer membrane anchoring of lipoproteins. Surprisingly, mLolB compensates for LolB function and supports E. coli growth, indicating that the N-terminal anchor is not essential for its function. Indeed, mLolB correctly localizes lipoproteins to either the inner or outer membrane depending on the sorting signal at the steady state. Furthermore, periplasmic mLolB enables the dissection of LolB function, namely lipoprotein binding, membrane targeting and lipoprotein anchoring. It mediates the transfer of lipoproteins from LolA to the outer membrane, but also the inner membrane and liposomes, indicating that mLolB exhibits no membrane preference and targets to phospholipids. Consequently, an outer membrane-specific lipoprotein is transiently mislocalized to the inner membrane when cells express only mLolB. LolB anchored to the outer membrane does not cause such mislocalization and is more active than mLolB. Phosphatidylethanolamine has been found to stimulate the mLolB-dependent membrane anchoring of lipoproteins. Taken together, these results indicate that lipoprotein binding, membrane targeting and membrane incorporation of lipoproteins are intrinsic functions of LolB. PMID- 19678845 TI - Growth of Frankia strains in leaf litter-amended soil and the rhizosphere of a nonactinorhizal plant. AB - The ability of Frankia strains to grow in the rhizosphere of a nonactinorhizal plant, Betula pendula, in surrounding bulk soil and in soil amended with leaf litter was analyzed 6 weeks after inoculation of pure cultures by in situ hybridization. Growth responses were related to taxonomic position as determined by comparative sequence analysis of nifH gene fragments and of an actinomycetes specific insertion in Domain III of the 23S rRNA gene. Phylogenetic analyses confirmed the basic classification of Frankia strains by host infection groups, and allowed a further differentiation of Frankia clusters within the Alnus host infection group. Except for Casuarina-infective Frankia strains, all other strains of the Alnus and the Elaeagnus host infection groups displayed growth in the rhizosphere of B. pendula, and none of them grew in the surrounding bulk soil that was characterized by a very low organic matter content. Only a small number of strains that all belonged to a distinct phylogenetic cluster within the Alnus host infection group grew in soil amended with ground leaf litter from B. pendula. These results demonstrate that saprotrophic growth of frankiae is a common trait for most members of the genus, and the supporting factors for growth (i.e. carbon utilization capabilities) varied with the host infection group and the phylogenetic affiliation of the strains. PMID- 19678844 TI - Use of stabilized luciferase-expressing plasmids to examine in vivo-induced promoters in the Vibrio cholerae vaccine strain CVD 103-HgR. AB - Live, attenuated Vibrio cholerae vaccines can induce potent immune responses after only a single oral dose. The strategy of harnessing these strains to present antigens from heterologous pathogens to the mucosal immune system shows great promise. To fully realize this possibility, V. cholerae strains must be created that stably express antigens in vivo in sufficient quantity to generate an immune response. In vivo-induced promoters have been shown to increase the stability and immunogenicity of foreign antigens expressed from multicopy plasmids. We report the construction of a series of genetically stabilized plasmids expressing luciferase as a heterologous protein from the following in vivo-induced promoters: V. cholerae P(argC), P(fhuC) and P(vca1008), and Salmonella enterica serovar Typhi P(ompC). We demonstrate that several of these expression plasmids meet two critical criteria for V. cholerae live vector vaccine studies. First, the plasmids are highly stable in the V. cholerae vaccine strain CVD 103-HgR at low copy number, in the absence of selective pressure. Second, real-time bioluminescent imaging (BLI) demonstrates inducible in vivo expression of the promoters in the suckling mouse model of V. cholerae colonization. Moreover, the use of BLI allows for direct quantitative comparison of in vivo expression from four different promoters at various time points. PMID- 19678847 TI - Calcium homeostasis and signaling in yeast cells and cardiac myocytes. AB - Calcium ions are the most ubiquitous and versatile signaling molecules in eukaryotic cells. Calcium homeostasis and signaling systems are crucial for both the normal growth of the budding yeast Saccharomyces cerevisiae and the intricate working of the mammalian heart. In this paper, we make a detailed comparison between the calcium homeostasis/signaling networks in yeast cells and those in mammalian cardiac myocytes. This comparison covers not only the components, structure and function of the networks but also includes existing knowledge on the measured and simulated network dynamics using mathematical models. Surprisingly, most of the factors known in the yeast calcium homeostasis/signaling network are conserved and operate similarly in mammalian cells, including cardiac myocytes. Moreover, the budding yeast S. cerevisiae is a simple organism that affords powerful genetic and genomic tools. Thus, exploring and understanding the calcium homeostasis/signaling system in yeast can provide a shortcut to help understand calcium homeostasis/signaling systems in mammalian cardiac myocytes. In turn, this knowledge can be used to help treat relevant human diseases such as pathological cardiac hypertrophy and heart failure. PMID- 19678846 TI - Seasonal and spatial variations in microbial community structure and diversity in the acid stream draining across an ongoing surface mining site. AB - This study examined the microbial community in an acidic stream draining across the Yun-Fu pyrite mine (Guangdong, China), where extremely acidic mine water is a persistent feature due to the intensive surface mining activities. Analysis of terminal restriction fragment length polymorphism (TRFLP) of 16S rRNA gene sequences showed that microbial populations varied spatially and seasonally and correlated with geochemical and physical conditions. After the stream moves from underground to the surface, the microbial community in the acidic water rapidly evolves into a distinct community close to that in the downstream storage pond. Comparisons of TRFLP peaks with sequenced clone libraries indicated that bacteria related to the recently isolated iron-oxidizer Ferrovum myxofaciens dominated the acidophilic community throughout the year except for the samples collected in spring from the storage pond, where Ferroplasma acidiphilum-like archaea represented the most abundant group. Acidithiobacillus ferrooxidans-affiliated organisms increased along the acid stream and remained common over the year, whereas Leptospirillum ferrooxidans-like bacteria were negligible or even not detected in the analyzed samples. The data indicate that changes in environmental conditions are accompanied by significant shifts in community structure of the prokaryotic assemblages at this opencast mining site. PMID- 19678848 TI - Critical role of RPI1 in the stress tolerance of yeast during ethanolic fermentation. AB - Stress tolerance of yeast Saccharomyces cerevisiae during ethanolic fermentation is poorly understood due to the lack of genetic screens and conventional plate assays for studying this phenotype. We screened a genomic expression library of yeast to identify gene(s) that, upon overexpression, would prolong the survival of yeast cells during fermentation, with the view to understand the stress response better and to use the identified gene(s) in strain improvement. The yeast RPI1 (Ras-cAMP pathway inhibitor 1) gene was identified in such a screen performed at 38 degrees C; introducing an additional copy of RPI1 with its native promoter helped the cells to retain their viability by over 50-fold better than the wild type (WT) parent strain, after 36 h of fermentation at 38 degrees C. Disruption of RPI1 resulted in a drastic reduction in viability during fermentation, but not during normal growth, further confirming the role of this gene in fermentation stress tolerance. This gene seems to improve viability by fortifying the yeast cell wall, because RPI1 overexpression strain is highly resistant to cell lytic enzyme zymolyase, compared with the WT strain. As the RPI1 overexpression strain substantially retains cell viability at the end of fermentation, the cells can be reused in the subsequent round of fermentation, which is likely to facilitate economical production of ethanol. PMID- 19678849 TI - Aspirin in the secondary prevention of cardiovascular disease: an update of the APTC meta-analysis. AB - We updated the 2002 Antiplatelet Trialists' Collaboration meta-analysis of antiplatelet therapy to assess the effects of aspirin alone in the secondary prevention of different types of thrombotic arterial disease. Results of randomized, placebo-controlled trials of aspirin in patients with confirmed cardiovascular disease were abstracted and synthesized by the Mantel-Haenszel method. We defined three cardiovascular disease groups according to the qualifying disease at entry: coronary artery disease (CAD), cerebrovascular disease (CRVD), and peripheral arterial disease (PAD). Results are given as odds ratios (OR) and 95% confidence intervals (95% CI). Compared with placebo, aspirin decreased significantly the risk of all-cause death in CAD and CRVD (OR = 0.80, 95% CI 0.75-0.86 and 0.91, 95% CI 0.85-0.98, respectively), and of vascular events in CAD, CRVD, and PAD (OR = 0.71, 95% CI 0.67-0.76, 0.87, 95% CI 0.82 0.93, and 0.50, 95% CI 0.29-0.88, respectively). The risk of non-fatal stroke was decreased in the CAD, CRVD, and PAD (OR = 0.64, 95% CI 0.50-0.83, 0.81, 95% CI 0.74-0.89, and 0.26, 95% CI 0.07-0.94, respectively). The risk of non-fatal myocardial infarction was decreased significantly in the CAD and CRVD (OR = 0.59, 95% CI 0.53-0.67, and 0.63, 95% CI 0.48-0.84, respectively), but not in the PAD (OR = 0.43, 95% CI 0.15-1.25). Aspirin nearly doubled the risk of major bleeds (OR = 1.87, 95% CI 1.51-2.32 for all clinical conditions). This meta-analysis confirms that aspirin decreases the risk of thrombotic events in patients with confirmed disease of the coronary, cerebrovascular, or peripheral artery beds. PMID- 19678850 TI - The comparison of the effects of anesthetic doses of ketamine, propofol, and etomidate on ischemia-reperfusion injury in skeletal muscle. AB - The fact that a considerable amount of clinical conditions suffering from ischemia-reperfusion injury (IRI) occur under general anesthesia has triggered researchers to focus on the effects of anesthetic drugs on IRI. Hence, the aim of this study was to compare the use of different anesthetic drugs in a skeletal IRI model. Tourniquet IRI method was performed and two experimental groups were established as sham-control and IRI group. Rats in each group were anesthetized either with thiopental, ketamine, propofol or etomidate. Malondialdehyde, superoxide dismutase, catalase, and glutathione peroxidase were measured in skeletal muscle via a spectrophotometer. Zinc, iron, copper, and selenium were evaluated by atomic absorption spectrophotometer. In rats anesthetized with thiopental (40 mg/kg, i.p.), malondialdehyde values in IRI group were higher and glutathion peroxidase levels were lower compared to sham-control group. However, superoxide dismutase and catalase activities were identical. On the other hand, while the level of zinc in IRI group attenuated, no differences in iron and copper values were determined. Rats anesthetized with ketamine (60 mg/kg), propofol (100 mg/kg), or etomidate (20 mg/kg) did not show increased malondialdehyde levels in comparison with control levels. While the drugs did not cause a distinction in the levels of superoxide dismutase, catalase, glutathion peroxidase, iron, and copper, zinc was in a lower level in IRI group compared to sham-control. In conclusion, ketamine, propofol, and etomidate, with anesthetic doses, denoted efficacious effects on IRI; hence the drugs might be preferred in certain operations with the risk of IRI. PMID- 19678851 TI - Droperidol and ondansetron in vitro electrophysiological drug interaction study. AB - Droperidol and ondansetron are potent anti-emetic agents which are often administered together. Although both drugs prolong QT interval in man by inhibition of Human Ether-a-go-go Related Gene-coded potassium channels, only droperidol was tested using more integrated experimental models. Therefore, we studied the effects of both compounds and their combination on action potentials (AP) of rabbit Purkinje fibers using conventional intracellular glass microelectrode. Purkinje fibers, driven at 1 Hz, were exposed to increasing concentrations (from 0.001 to 10 microm) of droperidol (n = 7) or ondansetron (n = 8) at 30 min intervals at 36.5 degrees C. Other fibers were exposed to a constant droperidol concentration (0.1 microm) alone (n = 7) or together with the same increasing concentrations of ondansetron (n = 6). Droperidol increased AP duration measured at 90% repolarization (APD90) in a concentration-dependent manner from 4.4 +/- 0.8% (mean +/- SEM) after 1 nm to a maximum of 158 +/- 72% after 1 microm. Ondansetron significantly increased APD90 by 5.3 +/- 2.1% at 100 nm up to 76 +/- 14% after 10 microm. Early after-depolarization occurred in 6/7 fibers exposed to droperidol and 1/8 fibers exposed to ondansetron. When given together, pure additive effects were observed. The concentrations that increased APD90 by 50% were 0.25 +/- 0.25 microm droperidol, 3.8 +/- 2.4 microm ondansetron and 1.5 +/- 0.8 microm ondansetron when given together with droperidol. Both ondansetron and droperidol prolong AP duration in Purkinje fibers, droperidol being 10 times more potent than ondansetron. Combination of ondansetron and droperidol exhibits an additive effect on AP duration. However, within clinically relevant concentrations, ondansetron does not further increase the AP prolongation caused by droperidol alone. PMID- 19678852 TI - Ellagic acid and its methyl-derivatives inhibit a newly found nitratase activity. AB - We have recently shown that low density lipoprotein (LDL) was able to denitrate albumin-bound 3-NO(2)-Tyr residues and to concomitantly release NO(3)(-) through a Ca(2+)-dependent process that has been ascribed to a specific protein structure. A lipophilic food component (gamma-tocopherol), which is easily loaded into LDL has been found to totally inhibit denitrating activity. We presently found that ellagic acid (EA) and its methylated derivatives, 4,4'O-methyl- and 3,3'O-methyl-ellagic acids (MeEA1 and MeEA2, respectively), amphipathic phenolic components of certain fruits and beverages, were also able to inhibit this activity, with a total inhibition for EA and a 60% inhibition for MeEA1 and MeEA2. EA exhibited the highest affinity for protein plasma, whereas a higher affinity of MeEA1 and MeEA2 (with MeEA1 > MeEA2) than EA was found for lipoprotein fractions, suggesting that the inhibition-driving property is protein affinity. As a result of this nitratase-inhibition property EA and its natural metabolite MeEA2 may have a beneficial role in special physiopathological conditions. PMID- 19678853 TI - Medical and non-medical direct costs of chronic low back pain in patients consulting primary care physicians in France. AB - A retrospective, observational, cohort study in primary care. To determine the total direct medical and non-medical cost of chronic low back pain (LBP) in France and its associated factors. Chronic LBP affects 5-10% of the population its burden in France is unknown. Ninety-eight randomly selected general practitioners included 796 adult patients with chronic LBP between October 2001 and December 2002. Direct costs due to physician visits, investigations, medications, hospitalizations, and other medical and non-medical resource use were collected for the 6 months prior to study visit. Costs both reimbursed and not by the French health insurance system were considered. Quality of life (QoL) and disease severity were measured using Short Form (SF)-8 and Roland-Morris disability questionnaire (RMDQ), respectively. Costs were updated to represent 2007 prices. Men represented 50.6% of the 796 patients, mean age was 53 +/- 11.3 years, and the duration of LBP was more than 1 year in 80.9% of patients. The total mean cost per patient over six months was 715.6 euro (95% CI: 644.2-797.8). Of these costs, 22.9% related to care provided by physiotherapists and allied specialists, 19.5% to medications, 17.4% to hospitalizations, 9.6% to investigations, and 12.5% to physician fees. In multivariate analysis, the factors associated with the cost of chronic LBP were disease severity (RMDQ score) and age of the patients. LBP is a disease that is both common and costly. PMID- 19678854 TI - Lipopolysaccharide binding of the mite allergen Der f 2. AB - Lipid-binding properties and/or involvement with host defense are often found in allergen proteins, implying that these intrinsic biological functions likely contribute to the allergenicity of allergens. The group 2 major mite allergens, Der f 2 and Der p 2, show structural homology with MD-2, the lipopolysaccharide (LPS)-binding component of the Toll-like receptor (TLR) 4 signalling complex. Elucidation of the ligand-binding properties of group 2 mite allergens and identification of interaction sites by structural studies are important to explore the relationship between allergenicity and biological function. Here, we report a ligand-fishing approach in which His-tagged Der f 2 was incubated with sonicated stable isotope-labelled Escherichia coli as a potential ligand source, followed by isolation of Der f 2-bound material by a HisTrap column and NMR analysis. We found that Der f 2 binds to LPS with a nanomolar affinity and, using fluorescence and gel filtration assays that LPS binds to Der f 2 in a molar ratio of 1 : 1. We mapped the LPS-binding interface of Der f 2 by NMR perturbation studies, which suggested that LPS binds Der f 2 between the two large beta sheets, similar to its binding to MD-2, the LPS-binding component of the innate immunity receptor TLR4. PMID- 19678855 TI - Effect of the F8 mutation c.1538-2A>T on pre-mRNA splicing. PMID- 19678856 TI - Vildagliptin therapy and hypoglycaemia in Muslim type 2 diabetes patients during Ramadan. AB - AIMS: To compare hypoglycaemic events, glycated haemoglobin (HbA(1c)) and changes in bodyweight in metformin-treated Muslim patients with type 2 diabetes receiving adjunctive treatment with vildagliptin or gliclazide during Ramadan fasting. METHODS: Data were collected from Muslim patients with type 2 diabetes attending primary care practices in North West London, whose HbA(1c) was > 8.5% despite treatment with metformin 2 g daily before Ramadan and who received gliclazide 160 mg twice daily (n = 26) or vildagliptin 50 mg twice daily (n = 26) in addition to metformin. Hypoglycaemic events, HbA(1c) and weight were recorded 2 weeks before and 10 days after the Ramadan fast. All patients received education about how to identify and manage hypoglycaemia during Ramadan. RESULTS: During Ramadan, at least one hypoglycaemic event (defined as blood glucose < 3.5 mmol/l with or without symptoms) was recorded in two patients receiving vildagliptin (7.7%) and 16 patients receiving gliclazide [61.5%; difference between groups -53.8%, 95% confidence interval (CI) -74.9 to -26.3, p < 0.001]. Vildagliptin was associated with a reduction in the mean number of hypoglycaemic events during Ramadan compared with before Ramadan, whereas gliclazide was associated with an increase (least squares mean difference between groups -0.66, 95% CI -1.20 to -0.13, p = 0.0168). Both gliclazide and vildagliptin were associated with similar reductions in HbA(1c) and a small, but insignificant, increase in weight. CONCLUSIONS: Appropriate treatment adjustments can lead to improved diabetes management during Ramadan, with avoidance of significant weight gain and improved glucose control without hypoglycaemia. The addition of vildagliptin to metformin therapy during Ramadan in Muslim patients with type 2 diabetes was associated with a reduction in the incidence of hypoglycaemia. PMID- 19678857 TI - Three-dimensional volume rendering of the ankle based on magnetic resonance images enables the generation of images comparable to real anatomy. AB - We have applied high-quality medical imaging techniques to study the structure of the human ankle. Direct volume rendering, using specific algorithms, transforms conventional two-dimensional (2D) magnetic resonance image (MRI) series into 3D volume datasets. This tool allows high-definition visualization of single or multiple structures for diagnostic, research, and teaching purposes. No other image reformatting technique so accurately highlights each anatomic relationship and preserves soft tissue definition. Here, we used this method to study the structure of the human ankle to analyze tendon-bone-muscle relationships. We compared ankle MRI and computerized tomography (CT) images from 17 healthy volunteers, aged 18-30 years (mean 23 years). An additional subject had a partial rupture of the Achilles tendon. The MRI images demonstrated superiority in overall quality of detail compared to the CT images. The MRI series accurately rendered soft tissue and bone in simultaneous image acquisition, whereas CT required several window-reformatting algorithms, with loss of image data quality. We obtained high-quality digital images of the human ankle that were sufficiently accurate for surgical and clinical intervention planning, as well as for teaching human anatomy. Our approach demonstrates that complex anatomical structures such as the ankle, which is rich in articular facets and ligaments, can be easily studied non-invasively using MRI data. PMID- 19678859 TI - The temporal course of mucoperiosteal flap revascularization at guided bone regeneration-treated implant sites: a pilot study. AB - AIMS: To investigate post-operative capillary density regeneration in healing mucoperiosteal flaps at guided bone regeneration-treated implant sites. MATERIAL AND METHODS: A non-invasive post-operative investigation was performed in 10 patients using orthogonal polarization spectral (OPS) imaging for assessment of capillary density during the course of mucoperiosteal flap wound healing for 6 weeks in patients receiving dental implants. RESULTS: The greatest increase in capillary regeneration occurred in the early wound-healing phase, during weeks 1 and 2, and recovery to baseline was achieved between weeks 4 and 5. A comparison of adjacent OPS measurements indicated that differences between the time point immediately following administration of local anaesthesia and directly post operatively ( p=0.002), between a directly post-operative time point and after 1 week (p=0.009), and between post-operative weeks 1 and 2 (p=0.036) were statistically significant. CONCLUSIONS: The early healing phase of mucoperiosteal flaps is characterized by rapid capillary regeneration. OPS imaging enabled the possibility to monitor and quantify the temporal development of mucoperiosteal flap revascularization following periodontal surgery. PMID- 19678860 TI - Association between periodontitis and anti-cardiolipin antibodies in Buerger disease. AB - AIM: Anti-cardiolipin (CL) antibodies can be induced in Buerger disease (BD), an inflammatory occlusive disorder affecting peripheral blood vessels, in response to bacteria bearing homology to the TLRVYK peptide of a phospholipid-binding plasma protein beta-2-glycoprotein I. TLRVYK homologies are present in Porphyromonas gingivalis (TLRIYT) and Treponema denticola (TLALYK). This study investigated the association between periodontal infection and anti-CL antibodies in BD patients. MATERIAL AND METHODS: Periodontal conditions were examined in 19 BD patients and 25 systemically healthy control subjects. All subjects were heavy smokers. Serum anti-CL, anti-TLRVYK, anti-TLRIYT, and anti-TLALYK antibodies were assessed using the enzyme-linked immunosorbent assay. RESULTS: BD patients had a significantly higher prevalence of periodontitis, more severe periodontal destruction and increased titres of serum anti-CL, anti-TLRVYK, anti-TLRIYT, and anti-TLALYK antibodies compared with healthy subjects. The levels of anti-CL antibodies positively correlated with those of the three anti-peptide antibodies. Anti-CL antibody titres were significantly associated with the percentage of sites with clinical attachment level >or=4 mm in BD patients. CONCLUSION: Elevated anti-CL antibody levels were associated with periodontal destruction in BD patients. Periodontopathic bacteria may serve as exogenous antigens that stimulate the anti-CL antibody production through molecular mimicry between the bacterial peptides and a host plasma protein. PMID- 19678861 TI - Clinical evaluation of a new collagen matrix (Mucograft prototype) to enhance the width of keratinized tissue in patients with fixed prosthetic restorations: a randomized prospective clinical trial. AB - AIM: The aim of this study was to test a new collagen matrix (CM) aimed to increase keratinized gingiva/mucosa when compared with the free connective tissue graft (CTG). MATERIAL AND METHODS: This randomized longitudinal parallel controlled clinical trial studied 20 patients with at least one location with minimal keratinized tissue (5 per hour and/or apnoea/hypopnoea index (AHI) > or =10 per hour, received an OA as treatment. BP was measured on three occasions; before treatment, after 3 months of treatment, and after 3 years of treatment. BP was measured with an electronic blood pressure monitor. The treatment effect of OA was measured after 3 months by repeated somnographic registration while the patient was wearing the OA. A treatment response was defined as AHI < 10; this was achieved in 25 of 29 patients (86%) at the 3-month evaluation. Significant reductions in blood pressure were attained between baseline and the 3-month evaluation (P < 0.001) and these changes remained at the 3-year follow-up in both systolic BP of -15.4 +/- 18.7 mm Hg and diastolic BP of -10.3 +/- 10.0 mm Hg. OA therapy reduced blood pressure in both a 3-month and a 3-year perspective in patients with OSA. PMID- 19678870 TI - Oral cancer cells with different potential of lymphatic metastasis displayed distinct biologic behaviors and gene expression profiles. AB - OBJECTIVE: Oral squamous cell carcinoma (OSCC) often spreads from the primary tumor to regional lymph nodes in the early stage. Better understanding of the biology of lymphatic spread of oral cancer cells is important for improving the survival rate of cancer patients. METHODS: We established the cell line LNMTca8113 by repeated injections in foot pads of nude mice, which had a much higher lymphatic metastasis rate than its parental cell line Tca8113. Then, we compared the biologic behaviors of cancer cells between them. Moreover, microarray-based expression profiles between them were also compared, and a panel of differential genes was validated using real-time-PCR. RESULTS: In contrast to Tca8113 cells, LNMTca8113 cells were more proliferative and resistant to apoptosis in the absence of serum, and had enhanced ability of inducing capillary like structures. Moreover, microarray-based expression profiles between them identified 1341 genes involved in cell cycle, cell adhesion, lymphangiogenesis, regulation of apoptosis, and so on. Some genes dedicating to the metastatic potential, including JAM2, TNC, CTSC, LAMB1, VEGFC, HAPLN1, ACPP, GDF9 and FGF11, were upregulated in LNMTca8113 cells. CONCLUSION: These results suggested that LNMTca8113 and Tca8113 cells were proper models for lymphatic metastasis study because there were differences in biologic behaviors and metastasis-related genes between them. Additionally, the differentially expressed gene profiles in cancer progression may be helpful in exploring therapeutic targets and provide the foundation for further functional validation of these specific candidate genes for OSCC. PMID- 19678871 TI - Small unilateral jaw gap variations: equilibrium changes, co-contractions and joint forces. AB - After complex prosthetic reconstructions, small differences in vertical distances between the left and right side of the jaw may occur during jaw closing, nevertheless providing bilateral tooth contacts in intercuspation by small deformations of the mandible. Their effects on the co-contraction of the masticatory muscles, the temporomandibular joint reaction forces, and the point of application of the resultant bite force vector in the maxillary occlusion plane - the so-called reduction point - have not been investigated, thus far simultaneously in one sample. The main goal of this study was to investigate variations of these measures in an experimental intercuspation simulated by one anterior and two posterior force transmission points. PMID- 19678872 TI - Intrinsic and extrinsic motivational orientations in the competitive context: an examination of person-situation interactions. AB - The current study examined Intrinsic Motivation Orientation and Extrinsic Motivation Orientation (Work Preference Inventory; Amabile, Hill, Hennessey, & Tighe, 1994) as potential trait-level moderators of the way Internet chess players responded to the intrinsic and extrinsic rewards of the chess games they played. On the basis of the defining characteristics of these 2 types of motivational orientations, we predicted that (a) Intrinsic Motivation Orientation would be associated with a stronger curvilinear relationship between challenge and enjoyment and (b) Extrinsic Motivation Orientation would be associated with a heightened affective responsivity to competitive outcome (i.e., winning vs. losing). Results supported the predictions. Implications of the findings are discussed. PMID- 19678873 TI - Recurrent personality dimensions in inclusive lexical studies: indications for a big six structure. AB - Previous evidence for both the Big Five and the alternative six-factor model has been drawn from lexical studies with relatively narrow selections of attributes. This study examined factors from previous lexical studies using a wider selection of attributes in 7 languages (Chinese, English, Filipino, Greek, Hebrew, Spanish, and Turkish) and found 6 recurrent factors, each with common conceptual content across most of the studies. The previous narrow-selection-based six-factor model outperformed the Big Five in capturing the content of the 6 recurrent wideband factors. Adjective markers of the 6 recurrent wideband factors showed substantial incremental prediction of important criterion variables over and above the Big Five. Correspondence between wideband 6 and narrowband 6 factors indicate they are variants of a "Big Six" model that is more general across variable-selection procedures and may be more general across languages and populations. PMID- 19678874 TI - Neuroticism and morning cortisol secretion: both heritable, but no shared genetic influences. AB - Neuroticism is widely used as an explanatory concept in etiological research of psychopathology. To clarify what neuroticism actually represents, we investigated the phenotypic and genetic relationship between neuroticism and the morning cortisol secretion. In the current classic twin study, 125 female twin pairs (74 monozygotic and 51 dizygotic pairs) participated. For each participant, 4 different neuroticism scores were available to calculate a neuroticism composite score that was used in the statistical analyses. The morning cortisol secretion was assessed by 4 salivary samples in the 1st hour after awakening. Significant genetic influences for the neuroticism composite score (55%), and each of the 4 cortisol samples (52%-69%) were found. There was no phenotypic or genotypic relationship between neuroticism and morning cortisol secretion. Although neuroticism and cortisol were both heritable traits, they did not share any genetic influences. PMID- 19678875 TI - Get a taste of your goals: promoting motive-goal congruence through affect-focus goal fantasy. AB - Studies show that motive-goal congruence is an important predictor of well-being (Baumann, Kaschel, & Kuhl, 2005; Brunstein, Schultheiss, & Grassmann, 1998). However, little is known about the factors that promote congruence between implicit motives and goals. Relying on McClelland's (1985) concept of implicit motives and the theory of fantasy realization (Oettingen, 1999), we postulated that goal fantasies focusing on motive-specific affective incentives promote motive-congruent goal setting. This hypothesis was tested in 3 experimental studies. In Study 1 (n=46) and Study 2 (n=48), participants were asked to select goals in a hypothetical scenario. In Study 3 (n=179), they rated their commitment to personal goals for their actual life situation. The results of all 3 studies supported our hypothesis that participants who focus on motive-specific affective incentives in their goal fantasies set their goals in line with their corresponding implicit motive dispositions. PMID- 19678876 TI - Narcissistic responding to ego threat: when the status of the evaluator matters. AB - Narcissists and nonnarcissists were insulted by high-status and low-status evaluators and were given an opportunity to self-protect with a comparative (evaluator derogation; Experiment 1) and noncomparative (inflated state self esteem; Experiments 1 and 2) strategy. Narcissists engaged in comparative self protection indiscriminately (i.e., derogating both low-status and high-status evaluators), whereas nonnarcissists showed some mercy to low-status evaluators. With regard to noncomparative protection, the findings were consistent across studies: Evaluator status interacted with narcissism such that narcissists engaged in noncomparative self-protection more than nonnarcissists when the evaluator was high, but not low, in status. Evaluator status and, more generally, source of feedback are worth serious consideration when untangling the intricacies and flexibility of narcissistic self-protection. PMID- 19678878 TI - Personal values and conflicting motivational forces in the context of imposed change. AB - Internal motivational conflicts that arise in the context of imposed change were investigated through a personal values perspective. It is suggested that in the context of imposed change different aspects of the same value dimension will tend to come in conflict. As demonstrated in two studies, this conflict is manifested in what at a surface level appears as a weak relationship between values and reaction to the change. In Study 1, a field study of 107 employees, individuals' dispositional resistance to change was controlled to disentangle the conflicting forces that employees experienced in response to a campus relocation. In Study 2, a laboratory study of 128 undergraduates, in addition to replicating the results of Study 1, the different motivational dynamics that exist in voluntary versus imposed change situations were demonstrated. PMID- 19678877 TI - Psychological needs as basic motives, not just experiential requirements. AB - Self-determination theory (SDT) posits 3 evolved psychological needs, for autonomy, competence, and relatedness. Considerable research has established that all 3 experiences are important for well-being. However, no SDT research has examined whether unmet needs have motivational force, an important criterion for establishing that certain experiences are indeed basic needs and motives (R. F. Baumeister & M. R. Leary, 1995). Three studies, using cross-sectional, experimental, and longitudinal methodologies, supply evidence that felt deficits in autonomy, competence, and relatedness arouse corresponding desires to acquire the missing experiences. However, a positive surfeit of felt-need satisfaction did not predict reduced desires for the corresponding needs. Implications for homeostatic, evolutionary, and humanistic perspectives upon basic psychological needs are discussed. PMID- 19678879 TI - The relevance of sexual responsiveness to sexual function in male stroke patients. AB - INTRODUCTION: Stroke may have negative consequences for the patients' quality of life, including sexual function. Whereas physical impairment will influence sexual positions and movement during sex, depression and medication may reduce sexual desire. So far, data on sexual dysfunction after stroke are scant. Although some support for physical as well as psychological explanations has been shown, further research to find the remedies for those patients with sexual problems after stroke is needed. The focus of the present study is on the identification of relevant psychological factors. AIM: The aim of this study was to study the impact of anxiety, depression, and sexual responsiveness on sexual function in male stroke patients. METHODS: Nineteen male stroke patients completed a number of self-report measures to assess psychological and sexual factors. MAIN OUTCOME MEASURES: Sexual function based on the International Index of Erectile Function, anxiety and depression based on the Symptom Checklist-90, and sexual responsiveness based on the Sexual Inhibition/Sexual Excitation Scale, including propensities for sexual excitation and sexual inhibition as a result of both performance failure and performance consequences, were assessed. RESULTS: Sexual excitation was positively related to sexual desire, whereas inhibition because of the threat of performance failure was negatively related to orgasmic function and sexual desire (P < 0.01). Patients with high levels of inhibition because of threat of performance failure were more likely to report low scores on overall sexual function than those with low levels. CONCLUSIONS: Although the statistical power is rather low, the results show the relevance of sexual responsiveness to sexual function in male stroke patients. The present study can be considered as a first step toward building a theoretical framework of relevant psychological and physical factors, which is needed to develop adequate interventions for those patients with sexual problems after stroke. PMID- 19678880 TI - Impaired cavernous reinnervation after penile nerve injury in rats with features of the metabolic syndrome. AB - INTRODUCTION: The metabolic syndrome is a cluster of cardiovascular risk factors that predispose toward the development of diseases such as diabetes. Erectile dysfunction (ED) is common in men with metabolic syndrome, but its etiology is poorly understood. Pro-erectile nitrergic nerves innervating penile erectile tissue are also susceptible to mechanical injury during pelvic surgical procedures, which can lead to sexual dysfunction. AIMS: The aims of this article are: (i) to examine erectile function in an experimental model of metabolic syndrome, the phosphoenolpyruvate carboxykinase (PEPCK)-overexpressing rat; and (ii) to study function and cavernous reinnervation after penile nerve crush injury, which permits regeneration, in transgenic rats. METHODS: We analyzed the density of noradrenergic and nitrergic nerves and performed organ bath pharmacology to assess neurogenic activity. MAIN OUTCOME MEASURES: By analyzing changes in neural structure, function, and pharmacologic responses of cavernous tissue after nerve crush injury, we were able to reveal neurologic deficits in rats with metabolic syndrome. RESULTS: Animals with features of metabolic syndrome did not develop notable changes in cavernous autonomic nerve density or nerve-evoked smooth muscle activity. However, regeneration of nitrergic nerves after crush injury in transgenic rats was impaired compared with injured controls. This was manifested as a deficit in axon regrowth and responses to axon activation. However, unlike injured controls, injured PEPCK-overexpressing rats did not develop a reduced maximal response to the nitric oxide (NO) donor, sodium nitroprusside. This suggests preserved NO responsiveness in tissues from rats with metabolic syndrome, despite impaired regeneration and return of function. CONCLUSIONS: This study revealed that rats with features of metabolic syndrome display impaired cavernous nerve regeneration after penile nerve injury, but the degree of functional impairment may be attenuated due to reduced plasticity of NO signaling. This reinnervation deficit may be of clinical relevance for understanding why ED persists in some (particularly aged) men after pelvic surgery. PMID- 19678881 TI - Sexual functioning in women with chronic pelvic pain: the role of anxiety and depression. AB - INTRODUCTION: Chronic pelvic pain (CPP) in women is a long lasting and often disabling condition. It seems reasonable to expect that as a result of the pain, extreme fatigue and/or emotional problems, women with CPP may report a variety of sexual problems. AIM: The present study investigated differences in the report of sexual problems in women with CPP compared with healthy controls, and whether the association of CPP with sexual problems was moderated or mediated by somatic and psychological factors as manifested in women suffering from CPP. METHOD: One hundred fifty-four women with CPP and 58 age-matched controls completed self report measures for sexual functioning, pain, physical impairment, anxiety, depression, and sexual and physical abuse. MAIN OUTCOME MEASURE: Golombok Rust Inventory of Sexual Satisfaction. RESULTS: Women with CPP reported higher levels of vaginistic complaints, sexual avoidance, nonsensuality and sexual dissatisfaction than healthy controls. Sexual problems were associated with anxiety, depression, and sexual abuse history but not with somatic factors as pain and physical impairment. Anxiety as well as depression, irrespective of the report of sexual abuse experiences, mediated the effect of CPP on sexual problems. Sexual abuse was a general predictor of sexual problems in both women with CPP and controls. CONCLUSIONS: Anxiety and depression constitute important factors in the evaluation of sexual problems in women with CPP. PMID- 19678883 TI - Sexual function in male patients undergoing treatment for renal failure: a prospective view. AB - INTRODUCTION: Chronic renal failure in males causes wide-ranging disturbances including sexual dysfunction. The percentage and progression of sexual dysfunction in patients entering a dialysis program require further evaluation. AIM: Our aim was to determine the ongoing effect of standard renal failure treatment on sexual function. METHODS: The sexual function was assessed prospectively, upon initiation of dialysis and every 10-12 months while on dialysis or after kidney transplantation. Participants were adult males with sexual partners. MAIN OUTCOME MEASURE: The semiquantitative standard International Index of Erectile Function questionnaire was used initially as a baseline and compared with all subsequent follow-up measures. RESULTS: Fifty-two patients fulfilled the eligibility criteria and completed the questionnaire. Of the 52 subjects, 25 were on hemodialysis and 27 were on peritoneal dialysis. Only 17.3% of participants were potent upon entry into the study. Of the rest, 71% was classified as suffering from severe erectile dysfunction (ED). Sexual desire appeared less affected when compared with other domains. Of the study participants, 67% expressed an interest in treatment for ED, but only 12% had ever received any such therapy. Follow-up ranged from 10 months to 48 months. After excluding kidney-transplanted patients, ED scores on entry and at four subsequent reassessments were almost identical and showed no significant statistical differences. Patients showed significant improvement in ED score after kidney transplantation, with scores remaining high for 20-36 months of follow-up, compared with pre-transplantation. CONCLUSIONS: This prospective study suggests that dialysis does not benefit sexual function, although a benefit was seen in a subset of men undergoing renal transplantation. We conclude that sexual function in men beginning dialysis should be assessed, and treatment for ED should be offered if appropriate. PMID- 19678882 TI - Metabolic syndrome and sexual (dys)function. AB - INTRODUCTION: The general worldwide increase in metabolic syndrome (MS) among most populations may result in more individuals with sexual dysfunction. AIM: To provide an update on clinical and experimental evidence regarding sexual dysfunction in patients with MS from both sexes and treatment modalities. METHODS: A comprehensive literature review was performed using MEDLINE with the MeSH terms and keywords for "metabolic syndrome,""obesity,""female sexual dysfunction,""erectile dysfunction,""androgen deficiency,""weight loss," and "bariatric surgery." MAIN OUTCOME MEASURES: To examine the data relating to sexual function in both men and women with MS, its relationship and the impact of treatment. RESULTS: The MS is strongly correlated with erectile dysfunction, hypogonadism (predictors of future development of MS), and female sexual dysfunction. Few studies have been addressed in the treatment of these dysfunctions in the special setting of MS, other than the observational effects on sexual function of individual risk factors correction. This can be a result of their understudied etiopathogeny. Nonsurgical weight loss has been shown to improve sexual function (with the mainstay on sedentarism prevention), whereas the efficacy of bariatric surgery in this respect, which has been suggested by some preliminary evidence, needs to be further confirmed by adequate clinical trials. CONCLUSION: As the global incidence of MS increases, more individuals may experience sexual dysfunction and a systematic evaluation should be emphasized in this patient population, in order to identify those who are in need of intervention. PMID- 19678884 TI - Phosphodiesterase type 5 inhibitors facilitate noncontact erections in male rats: site of action in the brain and mechanism of action. AB - INTRODUCTION: Orally active phosphodiesterase type 5 inhibitors (PDE5i), used in the treatment of erectile dysfunction, facilitate the relaxation of cavernous smooth muscle tissues by reducing the degradation of cyclic guanosine monophosphate. AIMS: The aims of this article were to determine whether PDE5i facilitate penile erection and male sexual behavior by acting also on the central nervous system and to investigate their mechanism of action at central level. METHODS: PDE5i (sildenafil, vardenafil, and tadalafil) given intraperitoneally (i.p.) (5 mg/kg and 10 mg/kg), intracerebroventricularly (i.c.v.) (10 microg and 50 microg), or into the ventral tegmental area (VTA) (10 microg) were tested in the noncontact erection test in male Sprague-Dawley rats screened for their ability to display or not display this sexual response. Extracellular dopamine was measured in the dialysate obtained from the nucleus accumbens by intracerebral microdialysis on injection of PDE5i into the VTA. MAIN OUTCOME MEASURES. Noncontact erections were counted after intraperitoneal, intracerebroventricular, or intra-VTA treatment with PDE5i. Extracellular dopamine was measured in the dialysate from the nucleus accumbens when sildenafil or vardenafil was given into the VTA. Results. PDE5i induced a significant increase of noncontact erections in male rats displaying this sexual response following intraperitoneal or intracerebroventricular administration at the highest dose tested. However, both doses significantly increased noncontact erections in male rats not displaying this sexual response. Similar results were found when PDE5i were injected into the caudal VTA. Noncontact erections increased concomitantly to a rise in extracellular dopamine in the dialysate from the nucleus accumbens. CONCLUSIONS: The results suggest that PDE5i may increase sexual arousal by acting in the central nervous system. This effect may be mediated (at least in part) by the activation of mesolimbic dopaminergic neurons. PMID- 19678885 TI - Effect of experimental hypothyroidism on glomerular filtration rate and plasma creatinine concentration in dogs. AB - BACKGROUND: Hypothyroidism affects renal function in a manner opposite the effects of hyperthyroidism. OBJECTIVE: To evaluate the effects of experimentally induced hypothyroidism on glomerular filtration rate (GFR) and basal plasma creatinine concentration in dogs. ANIMALS: Sixteen anestrous, female dogs. METHODS: Hypothyroidism was induced by administration of (131)I in 8 dogs, and 8 healthy euthyroid dogs acted as controls. Exogenous plasma creatinine clearance (an estimate of GFR) was measured in all dogs before (control period) and 43-50 weeks after induction of hypothyroidism (posttreatment period). Other pharmacokinetic parameters of creatinine were also determined. RESULTS: No significant difference was observed for basal plasma creatinine concentration and creatinine clearance between control and hypothyroid dogs in the control period. In the posttreatment period, mean + or - SD creatinine clearance in the hypothyroid group (2.13 + or - 0.48 mL/min/kg) was lower (P < .001) than that of the control group (3.20 + or - 0.42 mL/kg/min). Nevertheless, basal plasma creatinine concentrations were not significantly different between the hypothyroid and control groups (0.74 + or - 0.18 versus 0.70 + or - 0.08 mg/dL, respectively) because endogenous production of creatinine was decreased in hypothyroid dogs (22 + or - 3 versus 32 + or - 5 mg/kg/d, P=.001). CONCLUSION AND CLINICAL IMPORTANCE: Hypothyroidism causes a substantial decrease in GFR without altering plasma creatinine concentrations, indicating that GFR evaluation is needed to identify renal dysfunction in such patients. PMID- 19678886 TI - Optimal testing for thyroid hormone concentration after treatment with methimazole in healthy and hyperthyroid cats. AB - BACKGROUND: Methimazole suppresses thyroid hormone synthesis and is commonly used to treat feline hyperthyroidism. The degree of variation in thyroid hormone concentrations 24 hours after administration of methimazole and optimal time for blood sampling to monitor therapeutic efficacy have not been determined. OBJECTIVE: To assess thyroid hormone concentration variation in serum of normal and hyperthyroid cats after administration of methimazole. ANIMALS: Four healthy cats and 889 retrospectively acquired feline thyroid hormone profiles. METHODS: Crossover and retrospective studies. In the crossover study, healthy cats were treated with increasing doses of oral methimazole until steady state of thyroid suppression was achieved. Thyroid hormones and thyroid stimulating hormone (TSH) were serially and randomly monitored after methimazole. Paired t-tests and a 3 factor analysis of variance were used to determine differences between thyroid hormone concentrations in treated and untreated cats in the crossover study. Thyroid profiles from methimazole-treated hyperthyroid cats were retrieved from the Diagnostic Center for Population and Animal Health database and reviewed. Linear regression analysis evaluated relationships of dosage (mg/kg), dosing interval (q24h versus q12h), and time after methimazole to all thyroid hormone concentrations. RESULTS: All serum concentrations of thyroid hormones were significantly suppressed and TSH was significantly increased for 24 hours after administration of oral methimazole in healthy cats (P < .005). In hyperthyroid cats, there were no significant relationships between thyroid hormone concentrations and time postpill or dosing interval. CONCLUSIONS: Timing of blood sampling after oral methimazole administration does not appear to be a significant factor when assessing response to methimazole treatment. PMID- 19678887 TI - Effect of transvenous electrical cardioversion on plasma cardiac troponin I concentrations in horses with atrial fibrillation. AB - BACKGROUND: Whether electrical cardioversion of cardiac arrhythmias results in cardiomyocyte damage is unknown. OBJECTIVE: To describe effect of transvenous electrical cardioversion (TVEC) on plasma cardiac troponin I (cTnI) concentration in horses. ANIMALS: All horses presented to the Cornell University Hospital for Animals for cardioversion of atrial fibrillation between May 2006 and October 2008 were eligible for inclusion in the study. Owners of 14 horses elected for TVEC and each horse was then enrolled (16 procedures). METHODS: Prospective observational study measuring concentrations of plasma cTnI before and after TVEC. RESULTS: Median cTnI concentration increased from 0.045 ng/mL at baseline (range 0.0-0.20 ng/mL) to 0.11 ng/mL after TVEC (range 0.0-3.73 ng/mL) (P= .036). This increase was not associated with the number of shocks delivered, maximal energy delivered, cumulative energy delivered, chronicity of atrial fibrillation before cardioversion, or positioning of the pulmonary artery catheter. CONCLUSIONS: The increase in cTnI is unlikely to be clinically important. The increase might be correlated with persistent atrial dysfunction after TVEC, suggesting that a longer convalescent period after the procedure could be warranted. PMID- 19678888 TI - Temporal variability of ventricular arrhythmias in Boxer dogs with arrhythmogenic right ventricular cardiomyopathy. AB - BACKGROUND: Arrhythmogenic right ventricular cardiomyopathy (ARVC) is prevalent in the Boxer. There is little information on the temporal variability of ventricular arrhythmias within affected dogs. OBJECTIVE: To evaluate ambulatory electrocardiograms (AECG) from Boxers with ARVC for hourly variation in premature ventricular complexes (PVC) and heart rate (HR). ANIMALS: One hundred and sixty two Boxer dogs with ARVC. METHODS: Retrospective, observational study of 1,181 AECGs collected from Boxer dogs at The Ohio State University from 1997 to 2004 was evaluated. The proportion of depolarizations that were PVCs was compared across each hour of the day, during six 4-hour periods of day, to the time after AECG application, and to the maximum and minimum HR. RESULTS: A lower proportion of PVCs was noted during early morning (midnight to 0400 hours) as compared with the morning (0800-1200 hours) and late (1600-2000 hours) afternoon (P= .012). There was no increase in PVC proportion in the 1st hour after AECG application as compared with all other hours of the day (P= .06). There was poor correlation between maximum (rho= 0.19) and minimum (rho= 0.12) HR and PVC proportion. CONCLUSIONS AND CLINICAL IMPORTANCE: The likelihood of PVC occurrence in Boxer dogs with ARVC was relatively constant throughout the day, although slightly greater during the hours of 0800-1200 and 1600-2000. A biologically important correlation with HR was not apparent. The role of autonomic activity in the modulation of electrical instability in the Boxer with ARVC requires further study. PMID- 19678889 TI - Transcatheter atrial septal defect closure with the Amplatzer atrial septal occluder in 13 dogs: short- and mid-term outcome. AB - BACKGROUND: Transcatheter atrial septal defect (ASD) closure in the dog was first reported in 2005. OBJECTIVES: Describe the technique and both short- and mid-term outcome of transcatheter ASD closure with the Amplatzer atrial septal occluder (ASO). ANIMALS: Thirteen client-owned dogs with ASD. METHODS: Records of the initial 13 dogs in which transcatheter ASD closure was attempted at Texas A&M University were reviewed. RESULTS: All dogs had hemodynamically relevant septum secundum ASD. Two dogs had concurrent congenital abnormalities. ASOs were deployed in 13 dogs and released in 12. Eleven were released by a right jugular approach and 1 by a transatrial approach through a right lateral thoracotomy. Transthoracic echocardiographic estimates of ASD size were 14.0 + or - 5.4 mm (mean + or - 1 standard deviation) with a range of 7-22 mm. Accidental right atrial release occurred in 1 dog and embolization after release occurred in 2 dogs. Transcatheter ASD closure was successful in 10 dogs. Transthoracic color Doppler echocardiography the day after ASD closure indicated complete occlusion in 5 dogs, trivial to mild residual shunting in 4 dogs, and moderate residual shunting in 1 dog. Follow-up echocardiograms (mean of 12.4 + or - 7.4 months postprocedure) were available for 9 dogs. There was no residual ASD shunting in 6 dogs. In 3 of the 5 dogs with postoperative residual shunting it was judged to be decreased and hemodynamically unimportant relative to the dogs' postoperative evaluations. The mean length of event-free survival in the 10 dogs that underwent successful transcatheter ASD closure was 22.2 + or - 10.2 months. PMID- 19678890 TI - Retinol-binding protein in serum and urine of hyperthyroid cats before and after treatment with radioiodine. AB - BACKGROUND: Retinol-binding protein (RBP) is suggested as a clinically useful marker of renal function in cats. HYPOTHESIS: Serum and urinary RBP concentrations in hyperthyroid (HT) cats differ from those in healthy (H) cats; radioiodine ((131)I) treatment influences serum and urinary RBP concentrations in HT cats. ANIMALS: Ten HT and 8 H cats. METHODS: RBP concentration was evaluated in feline serum and urine samples from a prospective study. RESULTS: There was a significant (P= .003) difference in the urinary RBP/creatinine (uRBP/c) ratios of H (-) and untreated HT (1.4 + or - 1.5 x 10(-2) microg/mg) cats. Serum total thyroxine concentration (1.8 + or - 1.9 microg/dL, 24 weeks) and uRBP/c (0.6 + or - 1.0 x 10(-2) microg/mg, 24 weeks) decreased significantly (P < .001) in HT cats at all time points after treatment with (131)I, and these variables were significantly correlated with one another (r= 0.42, P= .007). Serum RBP concentrations from HT cats (199 + or - 86 microg/L) did not differ significantly (P= .98) from those of H cats (174 + or - 60) and did not change after treatment with (131)I (182 + or - 124 microg/L, P= .80). CONCLUSION AND CLINICAL IMPORTANCE: The presence of urinary RBP in HT cats is a potential marker of tubular dysfunction that is correlated to thyroid status, although it is independent of circulating RBP concentrations. The decreased uRBP/c combined with the absence of changes in serum RBP after treatment suggests that the suspected tubular dysfunction was partly reversible with treatment of (131)I. PMID- 19678891 TI - Remission of histiocytic ulcerative colitis in Boxer dogs correlates with eradication of invasive intramucosal Escherichia coli. AB - BACKGROUND: Historically, histiocytic ulcerative (HUC) (or granulomatous) colitis of Boxer dogs was considered an idiopathic immune-mediated disease with a poor prognosis. Recent reports of dramatic responses to enrofloxacin and the discovery of invasive Escherichia coli within the colonic mucosa of affected Boxer dogs support an infectious etiology. HYPOTHESIS: Invasive E. coli is associated with colonic inflammation in Boxer dogs with HUC, and eradication of intramucosal E. coli correlates with clinical and histologic remission. ANIMALS: Seven Boxer dogs with HUC. METHODS: Prospective case series. Colonic biopsies were obtained at initial evaluation in 7 dogs, and in 5 dogs after treatment with enrofloxacin. Biopsies were evaluated by standardized histopathology, and fluorescence in situ hybridization (FISH) with probes to eubacteria and E. coli. RESULTS: Intramucosal E. coli was present in colonic biopsies of 7/7 Boxers with HUC. Clinical response was noted in all dogs within 2 weeks of enrofloxacin (7 + or - 3.06 mg/kg q24 h, for 9.5 + or - 3.98 weeks) and was sustained in 6 dogs (median disease-free interval to date of 47 months, range 17-62). FISH was negative for E. coli in 4/5 dogs after enrofloxacin. E. coli resistant to enrofloxacin were present in the FISH-positive dog that relapsed. CONCLUSIONS AND CLINICAL RELEVANCE: The correlation between clinical remission and the eradication of mucosally invasive E. coli during treatment with enrofloxacin supports the causal involvement of E. coli in the development of HUC in susceptible Boxer dogs. A poor response to enrofloxacin treatment might be due to colonization with enrofloxacin-resistant E. coli. PMID- 19678893 TI - Factors affecting efficacy in patients with genotype 2 chronic hepatitis C treated by pegylated interferon alpha-2b and ribavirin: reducing drug doses has no impact on rapid and sustained virological responses. AB - Reducing the dose of drug affects treatment efficacy in pegylated interferon (Peg IFN) and ribavirin combination therapy for patients with hepatitis C virus (HCV) genotype 1. The aim of this study was to investigate the impact of drug exposure, as well as the baseline factors and the virological response on the treatment efficacy for genotype 2 patients. Two-hundred and fifty patients with genotype 2 HCV who were to undergo combination therapy for 24 weeks were included in the study, and 213 completed the treatment. Significantly more patients who achieved a rapid virological response (RVR), defined as HCV RNA negativity at week 4, achieved a sustained virological response (SVR) (92%, 122/133) compared with patients who failed to achieve RVR (48%, 38/80) (P < 0.0001). Multivariate logistic-regression analysis showed that only platelet counts [odds ratio (OR), 1.68; confidence interval (CI), 1.002-1.139] and RVR (OR, 11.251; CI, 5.184 24.419) were independently associated with SVR, with no correlation being found for the mean dose of Peg-IFN and ribavirin for RVR and SVR. Furthermore, in the stratification analysis of the timing of viral clearance, neither mean dose of Peg-IFN (P = 0.795) nor ribavirin (P = 0.649) affected SVR in each group. Among the patients with RVR, the lowest dose group of Peg-IFN (0.77 +/- 0.10 microg/kg/week) and ribavirin (6.9 +/- 0.90 mg/kg/day) showed 100% and 94% of SVR. Hence, RVR served as an important treatment predictor, and drug exposure had no impact on both SVR and RVR in combination therapy for genotype 2 patients. PMID- 19678892 TI - Combination chemotherapy with continuous L-asparaginase, lomustine, and prednisone for relapsed canine lymphoma. AB - BACKGROUND: The combination of lomustine, L-asparaginase, and prednisone (LAP) is an effective rescue treatment for canine lymphoma (LSA). In a previous study, we reported that remission was typically lost around the time L-asparaginase was discontinued. HYPOTHESIS: Use of L-asparaginase with each lomustine treatment will be well tolerated and efficacious as a rescue therapy for canine LSA. ANIMALS: Forty-eight client-owned dogs with cytologically confirmed multicentric LSA whose disease had relapsed after a cyclophosphamide, doxorubicin, vincristine, and prednisone-based chemotherapy protocol were included. METHODS: Lomustine was administered orally at 3-week intervals, concurrently with subcutaneous or intramuscular L-asparaginase for a total of 5 doses or until disease progression. Prednisone was administered at a tapering dose for the duration of the protocol. RESULTS: The overall response rate (ORR) for dogs treated with this protocol was 77%, with 65% achieving a complete response (CR). The median time to progression (TTP) was 70 days. Based on loose comparison, these findings are not significantly different from our previously reported historical control. The actual CCNU dosage administered did not affect response rate or remission duration. CONCLUSIONS/CLINICAL IMPORTANCE: These findings support previous data concluding that the LAP protocol is a viable rescue treatment option for dogs with LSA. However, results from this study suggest that continued use of L-asparaginase with each lomustine treatment does not significantly increase remission duration and toxicity appears greater. PMID- 19678894 TI - Low efficacy of entecavir therapy in adefovir-refractory hepatitis B patients with prior lamivudine resistance. AB - We determined the virologic response, incidence of entecavir resistance, and evolution of lamivudine and adefovir-resistant mutants during entecavir (ETV) therapy in adefovir-refractory patients with prior lamivudine resistance. Forty adefovir-refractory chronic hepatitis B patients with prior lamivudine resistance who had received entecavir for > or = 6 months were included and monitored for virologic response and entecavir resistance. Ten per cent of patients achieved HBV DNA < 50 copies/mL by PCR after 24 weeks of ETV therapy, and an initial virologic response was observed in 12 of 40 patients (30%). Higher pretreatment ALT (P = 0.039) and the presence of the rtL180M mutation (P = 0.038) were associated with an initial virologic response. During a mean follow-up of 11.4 months, four patients (10%) experienced virologic breakthrough, while ETV resistant mutants were detected in six patients (15%). YMDD and adefovir resistant mutants were detected in 57 and 35% of patients at baseline, respectively. At 48 weeks of therapy, 96 and 4% of patients had YMDD and adefovir resistant mutants, respectively. These data suggest an early development of ETV resistance and low antiviral response during ETV therapy in adefovir-refractory patients with prior lamivudine resistance. PMID- 19678895 TI - A mutational wrench in the HAMP gearbox. AB - HAMP domains communicate between input and output signalling elements in bacterial proteins. In the Tsr chemoreceptor, they convert axial movement of transmembrane helix 2 into changes in packing of the cytoplasmic kinase-control module (KCM). Zhou et al. suggest transmembrane helix 2 'tugs' on HAMP to destabilize x-da packing of the parallel four-helix bundle of the HAMP homodimer. Attractants would inhibit tugging. HAMP stability may be inversely related to stability of the a-d packing of the anti-parallel four-helix bundle of KCM, a relationship possibly facilitated by HAMP/KCM helical mismatch. The beauty of this idea lies in its simplicity and testability. PMID- 19678896 TI - Dynamic changes in the extracellular proteome caused by absence of a pleiotropic regulator AdpA in Streptomyces griseus. AB - In Streptomyces griseus, A-factor (2-isocapryloyl-3R-hydroxymethyl-gamma butyrolactone) triggers morphological development and secondary metabolism by inducing a pleiotropic transcriptional regulator AdpA. Extracellular proteome analysis of the wild-type and DeltaadpA strains grown to the end of the exponential phase in liquid minimal medium revealed that 38 secreted proteins, including many catabolic enzymes, such as protease, glycosyl hydrolase and esterase, were produced in an AdpA-dependent manner. Transcriptome analysis showed that almost all of these AdpA-dependent secreted proteins were regulated at the transcriptional level. In vitro AdpA-binding assays and determination of transcriptional start sites led to identification of 11 promoters as novel targets of AdpA. Viability staining revealed that some hyphae lysed during the exponential growth phase, which could explain the detection of 3 and 23 cytoplasmic proteins in the culture media of the wild-type and DeltaadpA strains respectively. In the wild-type strain, due to high protease activity in the culture medium, cytoplasmic proteins that leaked from dead cells seemed to be degraded and reused for the further growth. The existence of many AdpA-dependent (i.e. A-factor-inducible) secreted catabolic enzymes, which are likely involved in the assimilation of material that leaked from dead cells, reemphasizes the importance of A-factor in the morphological differentiation of S. griseus. PMID- 19678897 TI - Molecular characterization of putative vacuolar NHX-type Na(+)/H(+) exchanger genes from the salt-resistant tree Populus euphratica. AB - The vacuolar NHX-type Na(+)/H(+) exchangers play a key role in salt tolerance in plants. However, little is known about the Na(+)/H(+) exchangers in the salt resistant tree, Populus euphratica. In this study, we identified six putative vacuolar Na(+)/H(+) exchanger genes from P. euphratica, designated as PeNHX1-6. Real-time polymerase chain reaction indicated that the PeNHX1/3/6 transcripts were abundant compared with the other three PeNHX genes in the three tissues (roots, stems and leaves) examined. After NaCl treatment for 6 h, the transcript levels of PeNHX1-6 were upregulated in the roots. To address the function of PeNHX1-6, complementation studies were performed with the salt-sensitive yeast mutant strain R100, which lacks activity of the endosomal Na(+)/H(+) antiporter NHX1. The results showed that PeNHX1-6 compensates, at least in part, for the function of yeast NHX1. Moreover, PeNHX3 was targeted to the tonoplast when transiently expressed in onion. Together, these results suggest that PeNHX1-6 function as vacuolar Na(+)/H(+) exchangers and that PeNHX products play an important role in the salt resistance of P. euphratica. PMID- 19678898 TI - Methane emissions from six crop species exposed to three components of global climate change: temperature, ultraviolet-B radiation and water stress. AB - We examined the effects of temperature, ultraviolet-B (UVB) radiation and watering regime on aerobic methane (CH(4)) emission from six crops-faba bean, sunflower, pea, canola, barley and wheat. Plants were grown in controlled environment growth chambers under two temperature regimes (24/20 and 30/26 degrees C), three levels of UVB radiation [0 (zero), 5 (ambient) and 10 (enhanced) kJ m(-2) d(-1)] and two watering regimes (well watered and water stressed). A gas chromatograph with a flame ionization detector was used to measure CH(4) emission rates [ng g(-1) dry weight (DW) h(-1)] from detached fresh leaves of each species and attached leaves of pea plants. Plant growth [stem height, leaf area (LA) and aboveground dry matter (AG biomass)] and gas exchange [net CO(2) assimilation (A(N)), transpiration (E) and water use efficiency (WUE)] were also determined. We found that higher temperature, water stress and UVB radiation at the zero and enhanced levels significantly enhanced CH(4) emissions. Crop species varied in CH(4) emission, which was highest for pea and lowest for barley. Higher temperature and water stress reduced all growth parameters, whereas ambient and enhanced UVB decreased stem height but increased LA and AG biomass. Higher temperature decreased A(N) and WUE but increased E, whereas water stress decreased A(N) but increased E and WUE. Zero and enhanced UVB reduced A(N) and E. Growth and gas exchange varied with species. Overall, CH(4) emission was negatively correlated with stem height and AG biomass. We conclude that CH(4) emissions may increase under climatic stress conditions and this extra source might contribute to the 'greenhouse effect'. PMID- 19678900 TI - Use of a bridging autologous hepatic vein graft for extended right-liver transplantation. PMID- 19678901 TI - Should we perform heart retransplantation in early graft failure? AB - Cardiac retransplantation represents the gold standard treatment for a failing cardiac graft but the decision to offer the patient a second chance is often made difficult by both lack of donors and the ethical issues involved. The aim of this study was to evaluate whether retransplantation is a reasonable option in case of early graft failure. Between November 1985 and June 2008, 922 patients underwent cardiac transplantation at our Institution. Of these, 37 patients (4%) underwent cardiac retransplantation for cardiac failure resulting from early graft failure (n = 11) or late graft failure (acute rejection: n = 2, transplant-related coronary artery disease: n = 24). Survival at 1, 5 and 10 years of patients with retransplantation was 59%, 50% and 40% respectively. An interval between the first and the second transplantation of less than (n = 11, all in early graft failure) or more than (n = 26) 1 month was associated with a 1-year survival of 27% and 73%, and a 5-year survival of 27% and 65% respectively (P = 0.01). The long-term outcome of cardiac retransplantation is comparable with that of primary transplantation only in patients with transplant-related coronary artery disease. Early graft failure is a significant risk factor for survival after cardiac retransplantation and should be considered as an exclusion criteria. PMID- 19678899 TI - The COG complex, Rab6 and COPI define a novel Golgi retrograde trafficking pathway that is exploited by SubAB toxin. AB - Toxin trafficking studies provide valuable information about endogenous pathways of intracellular transport. Subtilase cytotoxin (SubAB) is transported in a retrograde manner through the endosome to the Golgi and then to the endoplasmic reticulum (ER), where it specifically cleaves the ER chaperone BiP/GRP78 (Binding immunoglobin protein/Glucose-Regulated Protein of 78 kDa). To identify the SubAB Golgi trafficking route, we have used siRNA-mediated silencing and immunofluorescence microscopy in HeLa and Vero cells. Knockdown (KD) of subunits of the conserved oligomeric Golgi (COG) complex significantly delays SubAB cytotoxicity and blocks SubAB trafficking to the cis Golgi. Depletion of Rab6 and beta-COP proteins causes a similar delay in SubAB-mediated GRP78 cleavage and did not augment the trafficking block observed in COG KD cells, indicating that all three Golgi factors operate on the same 'fast' retrograde trafficking pathway. SubAB trafficking is completely blocked in cells deficient in the Golgi SNARE Syntaxin 5 and does not require the activity of endosomal sorting nexins SNX1 and SNX2. Surprisingly, depletion of Golgi tethers p115 and golgin-84 that regulates two previously described coat protein I (COPI) vesicle-mediated pathways did not interfere with SubAB trafficking, indicating that SubAB is exploiting a novel COG/Rab6/COPI-dependent retrograde trafficking pathway. PMID- 19678902 TI - ADAP deficiency combined with costimulation blockade synergistically protects intestinal allografts. AB - Adhesion and degranulation promoting adapter protein (ADAP) plays an important role in T cell activation. ADAP deficiency was recently found to prolong heart graft survival in mice. We investigated the role of ADAP in intestinal transplantation and the synergistic effect of ADAP deficiency and Costimulation blockade (CB). T cell proliferation and cytotoxic T lymphocyte (CTL) activity were determined. MHC mismatched intestinal allografts was transplanted heterotopically. Anti-CD40L antibody was applied to the recipient. Upon stimulation with allogenic dendritic cells (DC), ADAP-deficient (ADAP-/-) T cells displayed impaired proliferative responses compared with that of wild-type (WT) T cells. In contrast, the CTL activity in ADAP-/- mice was comparable with that of WT mice. Rejection of intestinal allografts was ameliorated, but not prevented in ADAP-/- mice. Although CB alone was not sufficient to mitigate the rejection, the combination of CB and ADAP deficiency profoundly inhibited rejection. This was accompanied by less infiltration and activation of host lymphocytes in the gut associated lymphoid tissue of intestinal allografts. ADAP deficiency combined with CB protected the intestinal allografts synergistically. ADAP could be a novel target in the induction phase of the immune responses in organ transplantation. PMID- 19678903 TI - Comprehensive diagnostics for respiratory virus infections after transplantation or after potential exposure to swine flu A/H1N1: what else is out there? PMID- 19678905 TI - Macrophage migration inhibitory factor, infection, the brain, and corticosteroids. AB - Bacterial meningitis is a complex disorder in which injury is caused, in part, by the causative organism and, in part, by the host's own inflammatory response. Macrophage migration inhibitory factor (MIF) is a pro-inflammatory cytokine and a neuro-endocrine mediator that might play a role in pneumococcal meningitis. Here, we discuss the role of MIF in infection, the brain, and corticosteroids and conclude that experimental meningitis studies have to determine whether MIF is a potential target for adjunctive therapy in pneumococcal meningitis. PMID- 19678906 TI - Cell death in sepsis: a matter of how, when, and where. AB - Dysregulated cell death in several tissues is intimately involved in the pathogenesis of sepsis and contributes to multiple organ failure. Whether cell death during sepsis occurs by necrosis or apoptosis may depend on the cell type as well as the disease stage and is therefore a matter of intense debate. While lymphocyte apoptosis contributes to immunosuppression in sepsis, recent evidence suggests that necrosis of hepatocytes predominates in septic patients with liver dysfunction and correlates with poor survival. These distinct modes of cell death might have different consequences for the inflammatory response but are also critical for therapeutic interventions and the disease outcome. Understanding the complexity of death processes employing recently available serum biomarkers of cell death could lead to novel therapeutic approaches and assist in the steering of sepsis treatment. PMID- 19678907 TI - New data favouring that neurotrophins are of importance in arthritis. AB - Neurotrophins are important in inflammation. In an article in Arthritis Research & Therapy, Barthel and collaborators give new information on the existence of neurotrophin production in the synovial tissue of arthritic joints. These findings, together with other recent findings, stress that neurotrophins should be considered important factors in arthritis. This is reinforced by the facts that they are also produced by articular chondrocytes and that receptors for these are present in the synovial tissue and on chondrocytes. The importance of neurotrophins in joints should be further studied, including examinations on the efficacy of interfering with their effects in arthritis. PMID- 19678908 TI - Who is at increased risk for acute kidney injury following noncardiac surgery? AB - Abelha and colleagues evaluated the incidence and determinants of postoperative acute kidney injury (AKI) after major noncardiac surgery in patients with previously normal renal function. In this retrospective study of 1,166 patients with no previous renal insufficiency, who were admitted to a postsurgical intensive care unit (ICU) over a 2-year period, the incidence of AKI was 7.5%. Multivariate analysis identified American Society of Anesthesiologists physical status, Revised Cardiac Risk Index, high-risk surgery and congestive heart disease as preoperative AKI risk factors. AKI was an independent risk factor for hospital mortality (odds ratio = 3.12, 95% confidence interval = 1.41 to 6.93; P = 0.005), and was associated with higher severity of illness scores (Simplified Acute Physiology Score II and Acute Physiology and Chronic Health Evaluation II), longer ICU length of stay, higher ICU mortality, increased hospital mortality and higher mortality at 6-month follow up. Although the study design excluded 121 patients with significant preoperative renal insufficiency by design, the relatively crude serum creatinine cut-offs used certainly permitted inclusion of numerous patients with preoperative renal impairment. Accordingly, the study design failed to quantify the impact of preoperative renal impairment on risk and outcomes of perioperative AKI in noncardiac surgery, and this should be a goal of such studies in the future. Nonetheless, the study is an important addition to the literature in an under-studied population of patients at high risk for AKI. PMID- 19678910 TI - Linking genes to diseases: it's all in the data. AB - Genome-wide association analyses on large patient cohorts are generating large sets of candidate disease genes. This is coupled with the availability of ever increasing genomic databases and a rapidly expanding repository of biomedical literature. Computational approaches to disease-gene association attempt to harness these data sources to identify the most likely disease gene candidates for further empirical analysis by translational researchers, resulting in efficient identification of genes of diagnostic, prognostic and therapeutic value. Existing computational methods analyze gene structure and sequence, functional annotation of candidate genes, characteristics of known disease genes, gene regulatory networks, protein-protein interactions, data from animal models and disease phenotype. To date, a few studies have successfully applied computational analysis of clinical phenotype data for specific diseases and shown genetic associations. In the near future, computational strategies will be facilitated by improved integration of clinical and computational research, and by increased availability of clinical phenotype data in a format accessible to computational approaches. PMID- 19678911 TI - The human spermatozoon - a stripped down but refined machine. AB - A recent paper in BMC Developmental Biology describes the development of the annulus of the mouse sperm cell, but much remains to be learnt about sperm cells despite their importance in human fertility. PMID- 19678909 TI - Rheumatoid arthritis and smoking: putting the pieces together. AB - Besides atherosclerosis and lung cancer, smoking is considered to play a major role in the pathogenesis of autoimmune diseases. It has long been known that there is a connection between rheumatoid factor-positive rheumatoid arthritis and cigarette smoking. Recently, an important gene-environment interaction has been revealed; that is, carrying specific HLA-DRB1 alleles encoding the shared epitope and smoking establish a significant risk for anti-citrullinated protein antibody positive rheumatoid arthritis. We summarize how smoking-related alteration of the cytokine balance, the increased risk of infections (the possibility of cross reactivity) and modifications of autoantigens by citrullination may contribute to the development of rheumatoid arthritis. PMID- 19678913 TI - Novel genetic analysis in Behcet's disease. AB - Behcet's disease (BD) is characterized by oral and genital ulceration and is complicated by eye, skin, joint and central nervous system lesions. It has long been understood that BD has a strong genetic component, but to date the identified genes account for only around 30% of the risk for developing the disease, and the work has mostly been based on candidate gene analysis. In a recent report, Fei and coworkers presented the results of the first genome-wide analysis of patients with BD. These findings suggest new pathways for investigation in this complex disease. PMID- 19678912 TI - The determinants of susceptibility/resistance to adjuvant arthritis in rats. AB - Adjuvant arthritis (AA) serves as an excellent model for human rheumatoid arthritis. AA is readily inducible in certain rat strains, but not in others. Susceptibility/resistance to AA is determined by multiple factors. Among the genetic factors, both MHC and non-MHC genes contribute to arthritis susceptibility, and specific quantitative trait loci show association with the severity of the disease. Differential T-cell proliferative and cytokine responses, as well as antibody responses, to heat-shock proteins are evident when comparing AA-susceptible and AA-resistant rats. In addition, neuroendocrine factors and the housing environment can further modulate arthritis susceptibility/severity in particular rat strains. PMID- 19678914 TI - Central venous oxygen saturation and emergency intubation--another piece in the puzzle? AB - A recent multicentre observational study examined the effect of emergency intubation on central venous oxygen saturation (SCVo2) in critically ill patients. The main finding was that SCVo2 significantly increases 15 minutes after emergency intubation and institution of mechanical ventilation with 100% oxygen, especially in those patients with pre-intubation SCVo2 values <70%, regardless of whether these patients suffered from severe sepsis. However, in only one-quarter of this subgroup was the SCVo2 normalized to > or =70% solely by this intervention. In contrast, in patients with pre-intubation SCVo2 > or =70%, the SCVo2 failed to increase after intubation. A rise in SCVo2 can be expected when whole body oxygen extraction remains unchanged after intubation and ventilation with pure oxygen. PMID- 19678915 TI - Detecting volume responsiveness and unresponsiveness in intensive care unit patients: two different problems, only one solution. AB - Policies of fluid administration/restriction in critically ill patients have evolved over recent years. Abundant fluid resuscitation is encouraged during the early stage of severe sepsis. But a conservative fluid strategy is recommended in later stages, in particular when lungs are injured. Both strategies are risky if uncontrolled. Tests detecting volume unresponsiveness at any moment of fluid resuscitation or detecting volume unresponsiveness at any moment of fluid restriction would help to better assess the benefit/risk ratio of continuing such strategies. Measuring the short-term hemodynamic changes during passive leg raising can be reliably used for that purpose in both situations, even when patients are breathing spontaneously. PMID- 19678916 TI - Virus-mediated mRNA decay by hyperadenylation. AB - Degradation of cellular mRNAs during Kaposi's sarcoma-associated herpesvirus infection is associated with hyperadenylation of transcripts and a relocalization of cytoplasmic poly(A)-binding proteins to the nucleus. PMID- 19678917 TI - Infectious causes of cancer and their detection. AB - Molecular techniques for identifying pathogens associated with cancer continue to be developed, including one reported recently in BMC Medical Genomics. Identifying a causal infectious agent helps in understanding the biology of these cancers and can lead ultimately to the development of antimicrobial drugs and vaccines for their treatment and prevention. PMID- 19678918 TI - Pro/con debate: do the benefits of regionalized critical care delivery outweigh the risks of interfacility patient transport? AB - You are providing input in planning for critical care services to a large regional health authority. You are considering concentrating some critical care services into high-volume regional centres of excellence, as has been done in other fields of medicine. In your region, this would require several centres with differing levels of expertise that are geographically separated. Given there are inherent risks and time delays associated with interfacility patient transport, you debate whether these potential risks outweigh the benefits of regional centres of excellence. PMID- 19678919 TI - Intensity of renal replacement therapy in acute kidney injury: perspective from within the Acute Renal Failure Trial Network Study. AB - Determination of the optimal dose of renal replacement therapy in critically ill patients with acute kidney injury has been controversial. Questions have recently been raised regarding the design and execution of the US Department of Veterans Affairs/National Institutes of Health Acute Renal Failure Trial Network (ATN) Study, which demonstrated no improvement in 60-day all-cause mortality with more intensive management of renal replacement therapy. In the present article we present our rationale for these aspects of the design and conduct of the study, including our use of both intermittent and continuous modalities of renal support, our approach to initiation of study therapy and the volume management during study therapy. In addition, the article presents data on hypotension during therapy and recovery of kidney function in the perspective of other studies of renal support in acute kidney injury. Finally, we address the implications of the ATN Study results for clinical practice from the perspective of the study investigators. PMID- 19678921 TI - The free moment in walking and its change with foot rotation angle. AB - BACKGROUND: This investigation characterized the time-history pattern of the free moment (FM) during walking and, additionally, assessed whether walking with either an internally or externally rotated foot position altered the FM's time history. METHODS: Force plate and foot kinematic data were acquired simultaneously for 11 healthy subjects (6 males, 5 females) while walking at their self-selected comfortable speed in 3 foot rotation conditions (normal, internal and external). The FM was calculated and normalized by the product of each participant's body weight and height prior to extraction of peak FM, occurrence of peak FM in stance and net relative impulse. Differences in these values across foot rotation conditions were assessed using separate one-way, repeated measures analysis of variance and subsequent pair-wise comparisons. RESULTS: The average FM pattern during normal walking exhibits a biphasic shape: resisting inward rotation during approximately the first half of stance and outward rotation during the latter part of stance. While no differences in peak FM or net relative impulse were observed between the internal foot rotation condition and normal walking, the external foot rotation condition resulted in significantly greater peak FM and relative net impulse in comparison to normal walking. CONCLUSION: The differences in selected FM variables between normal walking and the external foot rotation condition are attributable to individual subject response to walking with an externally rotated foot. In this condition, some subjects displayed a FM pattern that was similar to that recorded during normal walking, while others displayed markedly larger FM patterns that are comparable in magnitude to those reported for running. The larger FM values in these latter subjects are speculated to be a result of excessive transverse plane body movements. Whilst further investigation is warranted regarding the FM time history characteristics during walking, our results indicate that the FM may provide useful information in assessment of gait. PMID- 19678920 TI - Mapping the human membrane proteome: a majority of the human membrane proteins can be classified according to function and evolutionary origin. AB - BACKGROUND: Membrane proteins form key nodes in mediating the cell's interaction with the surroundings, which is one of the main reasons why the majority of drug targets are membrane proteins. RESULTS: Here we mined the human proteome and identified the membrane proteome subset using three prediction tools for alpha helices: Phobius, TMHMM, and SOSUI. This dataset was reduced to a non-redundant set by aligning it to the human genome and then clustered with our own interactive implementation of the ISODATA algorithm. The genes were classified and each protein group was manually curated, virtually evaluating each sequence of the clusters, applying systematic comparisons with a range of databases and other resources. We identified 6,718 human membrane proteins and classified the majority of them into 234 families of which 151 belong to the three major functional groups: receptors (63 groups, 1,352 members), transporters (89 groups, 817 members) or enzymes (7 groups, 533 members). Also, 74 miscellaneous groups with 697 members were determined. Interestingly, we find that 41% of the membrane proteins are singlets with no apparent affiliation or identity to any human protein family. Our results identify major differences between the human membrane proteome and the ones in unicellular organisms and we also show a strong bias towards certain membrane topologies for different functional classes: 77% of all transporters have more than six helices while 60% of proteins with an enzymatic function and 88% receptors, that are not GPCRs, have only one single membrane spanning alpha-helix. Further, we have identified and characterized new gene families and novel members of existing families. CONCLUSION: Here we present the most detailed roadmap of gene numbers and families to our knowledge, which is an important step towards an overall classification of the entire human proteome. We estimate that 27% of the total human proteome are alpha-helical transmembrane proteins and provide an extended classification together with in-depth investigations of the membrane proteome's functional, structural, and evolutionary features. PMID- 19678922 TI - Corticomotor control of the genioglossus in awake OSAS patients: a transcranial magnetic stimulation study. AB - BACKGROUND: Upper airway collapse does not occur during wake in obstructive sleep apnea patients. This points to wake-related compensatory mechanisms, and possibly to a modified corticomotor control of upper airway dilator muscles. The objectives of the study were to characterize the responsiveness of the genioglossus to transcranial magnetic stimulation during respiratory and non respiratory facilitatory maneuvers in obstructive sleep apnea patients, and to compare it to the responsiveness of the diaphragm, with reference to normal controls. METHODS: Motor evoked potentials of the genioglossus and of the diaphragm, with the corresponding motor thresholds, were recorded in response to transcranial magnetic stimulation applied during expiration, inspiration and during maximal tongue protraction in 13 sleep apnea patients and 8 normal controls. MAIN RESULTS: In the sleep apnea patients: 1) combined genioglossus and diaphragm responses occurred more frequently than in controls (P < 0.0001); 2) the amplitude of the genioglossus response increased during inspiratory maneuvers (not observed in controls); 3) the latency of the genioglossus response decreased during tongue protraction (not observed in controls). A significant negative correlation was found between the latency of the genioglossus response and the apnea-hypopnea index; 4) the difference in diaphragm and genioglossus cortico motor responses during tongue protraction and inspiratory loading differed between sleep apnea and controls. CONCLUSION: Sleep apnea patients and control subjects differ in the response pattern of the genioglossus and of the diaphragm to facilitatory maneuvers, some of the differences being related to the frequency of sleep-related events. PMID- 19678923 TI - Presence of activating KRAS mutations correlates significantly with expression of tumour suppressor genes DCN and TPM1 in colorectal cancer. AB - BACKGROUND: Despite identification of the major genes and pathways involved in the development of colorectal cancer (CRC), it has become obvious that several steps in these pathways might be bypassed by other as yet unknown genetic events that lead towards CRC. Therefore we wanted to improve our understanding of the genetic mechanisms of CRC development. METHODS: We used microarrays to identify novel genes involved in the development of CRC. Real time PCR was used for mRNA expression as well as to search for chromosomal abnormalities within candidate genes. The correlation between the expression obtained by real time PCR and the presence of the KRAS mutation was investigated. RESULTS: We detected significant previously undescribed underexpression in CRC for genes SLC26A3, TPM1 and DCN, with a suggested tumour suppressor role. We also describe the correlation between TPM1 and DCN expression and the presence of KRAS mutations in CRC. When searching for chromosomal abnormalities, we found deletion of the TPM1 gene in one case of CRC, but no deletions of DCN and SLC26A3 were found. CONCLUSION: Our study provides further evidence of decreased mRNA expression of three important tumour suppressor genes in cases of CRC, thus implicating them in the development of this type of cancer. Moreover, we found underexpression of the TPM1 gene in a case of CRCs without KRAS mutations, showing that TPM1 might serve as an alternative path of development of CRC. This downregulation could in some cases be mediated by deletion of the TPM1 gene. On the other hand, the correlation of DCN underexpression with the presence of KRAS mutations suggests that DCN expression is affected by the presence of activating KRAS mutations, lowering the amount of the important tumour suppressor protein decorin. PMID- 19678924 TI - Development and initial validation of the Bedside Paediatric Early Warning System score. AB - INTRODUCTION: Adverse outcomes following clinical deterioration in children admitted to hospital wards is frequently preventable. Identification of children for referral to critical care experts remains problematic. Our objective was to develop and validate a simple bedside score to quantify severity of illness in hospitalized children. METHODS: A case-control design was used to evaluate 11 candidate items and identify a pragmatic score for routine bedside use. Case patients were urgently admitted to the intensive care unit (ICU). Control patients had no 'code blue', ICU admission or care restrictions. Validation was performed using two prospectively collected datasets. RESULTS: Data from 60 case and 120 control-patients was obtained. Four out of eleven candidate-items were removed. The seven-item Bedside Paediatric Early Warning System (PEWS) score ranges from 0-26. The mean maximum scores were 10.1 in case-patients and 3.4 in control-patients. The area under the receiver operating characteristics curve was 0.91, compared with 0.84 for the retrospective nurse-rating of patient risk for near or actual cardiopulmonary arrest. At a score of 8 the sensitivity and specificity were 82% and 93%, respectively. The score increased over 24 hours preceding urgent paediatric intensive care unit (PICU) admission (P < 0.0001). In 436 urgent consultations, the Bedside PEWS score was higher in patients admitted to the ICU than patients who were not admitted (P < 0.0001). CONCLUSIONS: We developed and performed the initial validation of the Bedside PEWS score. This 7 item score can quantify severity of illness in hospitalized children and identify critically ill children with at least one hours notice. Prospective validation in other populations is required before clinical application. PMID- 19678925 TI - Maternal attitudes and child-feeding practices: relationship with the BMI of Chilean children. AB - BACKGROUND: Chile has experienced the nutritional transition due to both social and economic progress. As a consequence, higher rates of overweight and obesity have been observed in children. In western countries, researchers have tried to determine pathways by which parents influence their children's eating behavior; up to now findings have been inconsistent. The objective of this study was to evaluate the cross-sectional and retrospective relationship between maternal attitudes and child-feeding practices and children's weight status in children who had been subject of an obesity prevention intervention for two years. METHODS: In 2006, for a cross-sectional study, a random sample of 232 children (125 girls, mean age 11.91 +/- 1.56 y and 107 boys mean age 11.98 +/- 1.51 y) was selected from three primary schools from a small city called Casablanca. Weight and height were determined to assess their nutritional status, using body mass index (BMI) z scores. Child-feeding practices and attitudes were determined cross sectionally in 2006, using the Child Feeding Questionnaire (CFQ). To analyze the relationship between trends in weight change and child-feeding practices and attitudes, BMI z scores of all the 232 children in 2003 were used. RESULTS: Cross sectionally, mothers of overweight children were significantly more concerned (P < 0.01) about their child's weight. Mothers of normal weight sons used significantly more pressure to eat (P < 0.05). Only in boys, the BMI z score was positively correlated with concern for child's weight (r = 0.28, P < 0.05) and negatively with pressure to eat (r = -0.21, P < 0.05). Retrospectively, the change in BMI z score between age 9 and 12 was positively correlated with concern for child's weight, but only in boys (r = 0.21, P < 0.05). Perceived child weight and concern for child's weight, explained 37% in boys and 45% in girls of the variance in BMI z score at age 12. CONCLUSION: Mothers of overweight children were more concerned with their children's weight; this indicated the Western negative attitude towards childhood overweight. None of the child-feeding practices were significantly correlated with a change in BMI z score. PMID- 19678926 TI - OnEX: Exploring changes in life science ontologies. AB - BACKGROUND: Numerous ontologies have recently been developed in life sciences to support a consistent annotation of biological objects, such as genes or proteins. These ontologies underlie continuous changes which can impact existing annotations. Therefore, it is valuable for users of ontologies to study the stability of ontologies and to see how many and what kind of ontology changes occurred. RESULTS: We present OnEX (Ontology Evolution EXplorer) a system for exploring ontology changes. Currently, OnEX provides access to about 560 versions of 16 well-known life science ontologies. The system is based on a three-tier architecture including an ontology version repository, a middleware component and the OnEX web application. Interactive workflows allow a systematic and explorative change analysis of ontologies and their concepts as well as the semi automatic migration of out-dated annotations to the current version of an ontology. CONCLUSION: OnEX provides a user-friendly web interface to explore information about changes in current life science ontologies. It is available at http://www.izbi.de/onex. PMID- 19678927 TI - Body image disturbance and surgical decision making in egyptian post menopausal breast cancer patients. AB - BACKGROUND: In most developing countries, as in Egypt; postmenopausal breast cancer cases are offered a radical form of surgery relying on their unawareness of the subsequent body image disturbance. This study aimed at evaluating the effect of breast cancer surgical choice; Breast Conservative Therapy (BCT) versus Modified Radical Mastectomy (MRM); on body image perception among Egyptian postmenopausal cases. METHODS: One hundred postmenopausal women with breast cancer were divided into 2 groups, one group underwent BCT and the other underwent MRM. Pre- and post-operative assessments of body image distress were done using four scales; Breast Impact of Treatment Scale (BITS), Impact of Event Scale (IES), Situational Discomfort Scale (SDS), and Body Satisfaction Scale (BSS). RESULTS: Preoperative assessment showed no statistical significant difference regarding cognitive, affective, behavioral and evaluative components of body image between both studied groups. While in postoperative assessment, women in MRM group showed higher levels of body image distress among cognitive, affective and behavioral aspects. CONCLUSION: Body image is an important factor for postmenopausal women with breast cancer in developing countries where that concept is widely ignored. We should not deprive those cases from their right of less mutilating option of treatment as BCT. PMID- 19678928 TI - Mechanisms of the action of povidone-iodine against human and avian influenza A viruses: its effects on hemagglutination and sialidase activities. AB - BACKGROUND: Influenza virus infection causes significant morbidity and mortality and has marked social and economic impacts throughout the world. The influenza surface glycoproteins, hemagglutinin (HA) and neuraminidase (NA), act cooperatively to support efficient influenza A virus replication and provide the most important targets for anti-influenza chemotherapy. In this study, povidone iodine (PVP-I), which has a broad-spectrum microbicidal property, was examined for its inhibitory effects against influenza virus infection in MDCK cells and the mechanisms of PVP-I action on HA and NA were revealed. RESULTS: Results obtained using a novel fluorescence- and chromogenic-based plaque inhibition assay showed that 1.56 mg/ml PVP-I inhibited infections in MDCK cells of human (8 strains) and avian (5 strains) influenza A viruses, including H1N1, H3N2, H5N3 and H9N2, from 23.0-97.5%. A sialidase inhibition assay revealed that PVP-I inhibited N1, N2 and N3 neuraminidases with IC50 values of 9.5-212.1 microg/ml by a mixed-type inhibition mechanism. Receptor binding inhibition and hemagglutinin inhibition assays indicated that PVP-I affected viral hemagglutinin rather than host-specific sialic acid receptors. CONCLUSION: Mechanisms of reduction of viral growth in MDCK cells by PVP-I involve blockade of viral attachment to cellular receptors and inhibition of viral release and spread from infected cells. Therefore, PVP-I is useful to prevent infection and limit spread of human and avian influenza viruses. PMID- 19678930 TI - Profile of hematological abnormalities of Indian HIV infected individuals. AB - BACKGROUND: Hematological abnormalities are a common complication of HIV infection. These abnormalities increase as the disease advances. Bone marrow abnormalities occur in all stages of HIV infection. METHODS: Two hundred HIV infected individual were screened for hematological abnormalities from March 2007 March 2008. Absolute CD4 cell count analysis was carried out by flowcytometry. Depending on the results of the primary screening further investigations were performed, like iron studies, hemolytic work up, PNH work up and bone marrow evaluation. Other investigations included coagulation profile, urine analysis, blood culture (bacterial, fungal, mycobacterial), serology for Epstein Barr virus (EBV), Cytomegalovirus (CMV), Hepatitis B and C, and Parvo B19 infection. RESULTS: The most common hematological abnormality was anemia, seen in 65.5% (131/200) patients. Iron deficiency anemia was seen in 49.2% (/200) cases while anemia of chronic disease occurred in 50.7% (/200) cases. Bone marrow evaluation was carried out in 14 patients out of which staging marrow was performed in 2 cases of non-Hodgkin's lymphoma (NHL) and did not show any bone marrow infiltration. In remaining 12 cases bone marrow was done for evaluation of pancytopenia. Among patients with pancytopenia 50% (6/12) showed granulomas (4 were positive for AFB, 2 were positive for fungal cryptococci), 25% (3/12) showed hemophagocytosis. There was a strong negative correlation between anemia and CD4 counts in this study. Thrombocytopenia was seen in 7% (14/200) cases and had no significant correlation with CD4 counts. No patient had absolute neutrophil count (ANC) < 800 cells/microL. No case of coagulation abnormalities was found. CONCLUSION: Anemia in HIV patients can be a good clinical indicator to predict and access the underlying immune status. Patients should be investigated for hematological manifestations and appropriate steps should be taken to identify and treat the reversible factors. PMID- 19678929 TI - Tumor biology and cancer therapy - an evolving relationship. AB - The aim of palliative chemotherapy is to increase survival whilst maintaining maximum quality of life for the individual concerned. Although we are still continuing to explore the optimum use of traditional chemotherapy agents, the introduction of targeted therapies has significantly broadened the therapeutic options. Interestingly, the results from current trials put the underlying biological concept often into a new, less favorable perspective. Recent data suggested that altered pathways underlie cancer, and not just altered genes. Thus, an effective therapeutic agent will sometimes have to target downstream parts of a signaling pathway or physiological effects rather than individual genes. In addition, over the past few years increasing evidence has suggested that solid tumors represent a very heterogeneous group of cells with different susceptibility to cancer therapy. Thus, since therapeutic concepts and pathophysiological understanding are continuously evolving a combination of current concepts in tumor therapy and tumor biology is needed. This review aims to present current problems of cancer therapy by highlighting exemplary results from recent clinical trials with colorectal and pancreatic cancer patients and to discuss the current understanding of the underlying reasons. PMID- 19678932 TI - Physicians' working conditions and job satisfaction: does hospital ownership in Germany make a difference? AB - BACKGROUND: A growing number of German hospitals have been privatized with the intention of increasing cost effectiveness and improving the quality of health care. Numerous studies investigated what possible qualitative and economic consequences these changes issues might have on patient care.However, little is known about how this privatization trend relates to physicians' working conditions and job satisfaction. It was anticipated that different working conditions would be associated with different types of hospital ownership. To that end, this study's purpose is to compare how physicians, working for both public and privatized hospitals, rate their respective psychosocial working conditions and job satisfaction. METHODS: The study was designed as a cross sectional comparison using questionnaire data from 203 physicians working at German hospitals of different ownership types (private for-profit, public and private nonprofit). RESULTS: The present study shows that several aspects of physicians' perceived working conditions differ significantly depending on hospital ownership. However, results also indicated that physicians' job satisfaction does not vary between different types of hospital ownership. Finally, it was demonstrated that job demands and resources are associated with job satisfaction, while type of ownership is not. CONCLUSION: This study represents one of a few studies that investigate the effect of hospital ownership on physicians work situation and demonstrated that the type of ownership is a potential factor accounting for differences in working conditions. The findings provide an informative basis to find solutions improving physicians' work at German hospitals. PMID- 19678931 TI - Biological mechanisms of disease and death in Moscow: rationale and design of the survey on Stress Aging and Health in Russia (SAHR). AB - BACKGROUND: Prior research has revealed large differences in health and mortality across countries, socioeconomic groups, and individuals. Russia experiences one of the world's highest levels of all-cause and cardiovascular mortality, great mortality differences within the population, and a heavy burden of ill health. Psychological stress has been suggested as a likely explanation of health loss and premature death in Russia and Eastern Europe. However, physiological mechanisms connecting stress with health in Russia remain unclear since existing epidemiological data are scarce and limited to conventional risk factors. METHOD AND DESIGN: The survey on Stress Aging and Health in Russia (SAHR) is addressing this knowledge gap by collecting an unusually rich database that includes a wide range of reported information, physical and cognitive health outcomes, and biomarkers in a sample of Muscovite men and women aged 55 and older. The total planned sample size is 2,000 individuals. The sample was randomly selected from epidemiological cohorts formed in Moscow between the mid-1970s and the 1990s and from medical population registers. The baseline data collection was carried out from December 2006 to June 2009. Interviews and medical tests were administered at hospital or at home according to standardized protocol. Questionnaire information includes health, socio-demographic characteristics, economic well being, cognitive functioning, and batteries on stress and depression. Biomarkers include anthropometry, grip strength, resting ECG, conventional cardiovascular factors of risk such as lipid profile and blood pressure, and other biochemical parameters such as those related to inflammation, glucose and insulin resistance, coagulation, fibrinolysis, and stress hormones. In addition to these measurements, SAHR includes dynamic biomarkers provided by 24-hour ECG (Holter) monitoring. This method continuously registers the beat-to-beat heart rate in naturalistic conditions without restrictions on normal daily activities. It provides information about heart functioning, including heart rate variability and ischemic and arrhythmic events.Re-examination of the study subjects will be conducted in 2009-2011 and will focus on health, functional status, economic conditions, behaviors, and attitudes towards aging. The subjects are also followed up for mortality and non-fatal health events. DISCUSSION: The SAHR will produce a valuable set of established and novel biomarkers combined with self reported data for the international research community and will provide important insights into factors and biological mechanisms of mortality and health losses in Russia. PMID- 19678936 TI - Comparative gene expression profiles between heterotic and non-heterotic hybrids of tetraploid Medicago sativa. AB - BACKGROUND: Heterosis, the superior performance of hybrids relative to parents, has clear agricultural value, but its genetic control is unknown. Our objective was to test the hypotheses that hybrids expressing heterosis for biomass yield would show more gene expression levels that were different from midparental values and outside the range of parental values than hybrids that do not exhibit heterosis. RESULTS: We tested these hypotheses in three Medicago sativa (alfalfa) genotypes and their three hybrids, two of which expressed heterosis for biomass yield and a third that did not, using Affymetrix M. truncatula GeneChip arrays. Alfalfa hybridized to approximately 47% of the M. truncatula probe sets. Probe set signal intensities were analyzed using MicroArray Suite v.5.0 (MAS) and robust multi-array average (RMA) algorithms. Based on MAS analysis, the two heterotic hybrids performed similarly, with about 27% of genes showing differential expression among the parents and their hybrid compared to 12.5% for the non-heterotic hybrid. At a false discovery rate of 0.15, 4.7% of differentially expressed genes in hybrids (approximately 300 genes) showed nonadditive expression compared to only 0.5% (16 genes) in the non-heterotic hybrid. Of the nonadditively expressed genes, approximately 50% showed expression levels that fell outside the parental range in heterotic hybrids, but only one of 16 showed a similar profile in the non-heterotic hybrid. Genes whose expression differed in the parents were three times more likely to show nonadditive expression than genes whose parental transcript levels were equal. CONCLUSION: The higher proportions of probe sets with expression level that differed from the parental midparent value and that were more extreme than either parental value in the heterotic hybrids compared to a non-heterotic hybrid were also found using RMA. We conclude that nonadditive expression of transcript levels may contribute to heterosis for biomass yield in alfalfa. PMID- 19678937 TI - Integrating microarray analysis and the soybean genome to understand the soybeans iron deficiency response. AB - BACKGROUND: Soybeans grown in the upper Midwestern United States often suffer from iron deficiency chlorosis, which results in yield loss at the end of the season. To better understand the effect of iron availability on soybean yield, we identified genes in two near isogenic lines with changes in expression patterns when plants were grown in iron sufficient and iron deficient conditions. RESULTS: Transcriptional profiles of soybean (Glycine max, L. Merr) near isogenic lines Clark (PI548553, iron efficient) and IsoClark (PI547430, iron inefficient) grown under Fe-sufficient and Fe-limited conditions were analyzed and compared using the Affymetrix GeneChip Soybean Genome Array. There were 835 candidate genes in the Clark (PI548553) genotype and 200 candidate genes in the IsoClark (PI547430) genotype putatively involved in soybean's iron stress response. Of these candidate genes, fifty-eight genes in the Clark genotype were identified with a genetic location within known iron efficiency QTL and 21 in the IsoClark genotype. The arrays also identified 170 single feature polymorphisms (SFPs) specific to either Clark or IsoClark. A sliding window analysis of the microarray data and the 7X genome assembly coupled with an iterative model of the data showed the candidate genes are clustered in the genome. An analysis of 5' untranslated regions in the promoter of candidate genes identified 11 conserved motifs in 248 differentially expressed genes, all from the Clark genotype, representing 129 clusters identified earlier, confirming the cluster analysis results. CONCLUSION: These analyses have identified the first genes with expression patterns that are affected by iron stress and are located within QTL specific to iron deficiency stress. The genetic location and promoter motif analysis results support the hypothesis that the differentially expressed genes are co-regulated. The combined results of all analyses lead us to postulate iron inefficiency in soybean is a result of a mutation in a transcription factor(s), which controls the expression of genes required in inducing an iron stress response. PMID- 19678933 TI - Impact of cigarette smoking on the relationship between body mass index and coronary heart disease: a pooled analysis of 3264 stroke and 2706 CHD events in 378579 individuals in the Asia Pacific region. AB - BACKGROUND: Elevated levels of body mass index (BMI) and smoking are well established lifestyle risk factors for coronary heart disease (CHD) and stroke. If these two risk factors have a synergistic relationship, rigorous lifestyle modification may contribute to greater reduction in cardiovascular burden than previously expected. METHODS: A pooled analysis of individual participant data from 38 cohorts, involving 378,579 participants. Hazards ratios (HRs) and 95% confidence intervals (CIs) for BMI by cigarette smoking status were estimated using Cox proportional hazard models. RESULTS: During a mean follow-up of 3.8 years, 2706 CHD and 3264 strokes were recorded. There was a log-linear, positive relationship of BMI with CHD and stroke in both smokers and non-smokers with evidence of a synergistic effect of smoking on the association between BMI and CHD only: HRs (95% CIs) associated with a 2 kg/m2 higher BMI were 1.13 (1.10 1.17) in current smokers and 1.09 (1.06-1.11) in non-smokers (p-value for interaction=0.04). CONCLUSION: Smoking amplifies the positive association between BMI and CHD but not stroke. If confirmed, these results suggest that effective strategies that target smoking cessation and weight loss are likely to have a greater impact than anticipated on reducing the burden of CHD. PMID- 19678934 TI - Reduction of lipoxidative load by secretory phospholipase A2 inhibition protects against neurovascular injury following experimental stroke in rat. AB - BACKGROUND: In animal models, ischemia reperfusion (IR) injury triggers membrane lipid degradation and accumulation of lipoxidative exacerbations in neurovascular unit, leading to blood brain barrier (BBB) damage and neurologic deficits. In this study, we investigated whether impeding membrane lipid breakdown by inhibiting secretory phospholipase A2 (sPLA2) activity reduces BBB leakage, leading to neuroprotection and functional recovery. METHODS: Focal cerebral IR injury was induced by middle cerebral artery occlusion (MCAO) in adult male rats. A sPLA2 inhibitor, 7,7-dimethyleicosadienoic acid (DEDA), was administered following IR injury. DEDA-treated animals were compared with vehicle-treated in terms of BBB leakage, edema, infarct volume, and neurological deficit. Membrane lipid degradation and the expression/activity of sPLA2 were also assessed. The role of one of the sPLA2 products, arachidonic acid (AA), on the morphology of the differentiated neuronal cell PC12 was examined by light microscopy. RESULTS: Treatment with DEDA after IR injury not only reduced BBB leakage but also decreased infarct volume and improved neurologic function. The treatment attenuated both the activity of sPLA2 and the levels of sPLA2-derived oxidized products. The metabolites of lipid oxidation/peroxidation, including the protein carbonyl, were reduced as well. The treatment also restored the levels of glutathione, indicating attenuation of oxidative stress. In vitro treatment of PC12 cells with DEDA did not restore the AA-mediated inhibition of neurite formation and the levels of glutathione, indicating that effect of DEDA is up stream to AA release. CONCLUSION: sPLA2-derived oxidative products contribute to significant neurovascular damage, and treatment with sPLA2 inhibitor DEDA ameliorates secondary injury by reducing exacerbations from lipoxidative stress. PMID- 19678939 TI - Exogenous Cx43 expression decrease cell proliferation rate in rat hepatocarcinoma cells independently of functional gap junction. AB - BACKGROUND: Gap junction intercellular communication (GJIC) is considered to play a role in the regulation of homeostasis because it regulates important processes, such as cell proliferation and cell differentiation. A reduced or lost GJIC capacity has been observed in solid tumors and studies have demonstrated that GJIC restoration in tumor cells contribute to reversion of the transformed phenotype. This observation supports the idea that restoration of the functional channel is essential in this process. However, in the last years, reports have proposed that just the increase in the expression of specific connexins can contribute to reversion of the malign phenotype in some tumor cells. In the present work, we studied the effects of exogenous Connexin 43 (Cx43) expression on the proliferative behavior and phenotype of rat hepatocarcinoma cells. RESULTS: The exogenous Cx43 did not increase GJIC capacity of transfected cells, but it was critical to decrease the cell proliferation rate as well as reorganization of the actin filaments and cell flattening. We also observed more adhesion capacity to substrate after Cx43 transfection. CONCLUSION: Cx43 expression leads to a decrease of the growth of the rat hepatocellular carcinoma cells and it contributes to the reversion of the transformed phenotype. These effects were independent of the GJIC and were probably associated with the phosphorylation pattern changes and redistribution of the Cx43 protein. PMID- 19678938 TI - The Ras guanine nucleotide exchange factor RasGRF1 promotes matrix metalloproteinase-3 production in rheumatoid arthritis synovial tissue. AB - INTRODUCTION: Fibroblast-like synoviocytes (FLS) from rheumatoid arthritis (RA) patients share many similarities with transformed cancer cells, including spontaneous production of matrix metalloproteinases (MMPs). Altered or chronic activation of proto-oncogenic Ras family GTPases is thought to contribute to inflammation and joint destruction in RA, and abrogation of Ras family signaling is therapeutic in animal models of RA. Recently, expression and post translational modification of Ras guanine nucleotide releasing factor 1 (RasGRF1) was found to contribute to spontaneous MMP production in melanoma cancer cells. Here, we examine the potential relationship between RasGRF1 expression and MMP production in RA, reactive arthritis, and inflammatory osteoarthritis synovial tissue and FLS. METHODS: Expression of RasGRF1, MMP-1, MMP-3, and IL-6 was detected in synovial tissue by immunohistochemistry and stained sections were evaluated by digital image analysis. Expression of RasGRF1 in FLS and synovial tissue was also assessed by immunoblotting. Double staining was performed to detect proteins in specific cell populations, and cells producing MMP-1 and MMP 3. RasGRF1 expression was manipulated in RA FLS by cDNA transfection and gene silencing, and effects on MMP-1, TIMP-1, MMP-3, IL-6, and IL-8 production measured by ELISA. RESULTS: Expression of RasGRF1 was significantly enhanced in RA synovial tissue, and detected in FLS and synovial macrophages in situ. In cultured FLS and synovial biopsies, RasGRF1 was detected by immunoblotting as a truncated fragment lacking its negative regulatory domain. Production of MMP-1 and MMP-3 in RA but not non-RA synovial tissue positively correlated with expression of RasGRF1 and co-localized in cells expressing RasGRF1. RasGRF1 overexpression in FLS induced production of MMP-3, and RasGRF1 silencing inhibited spontaneous MMP-3 production. CONCLUSIONS: Enhanced expression and post translational modification of RasGRF1 contributes to MMP-3 production in RA synovial tissue and the semi-transformed phenotype of RA FLS. PMID- 19678940 TI - The search for valved conduit tissue grafts for adults (>22 mm): an ultrasonographic study of jugular vein diameters of horses and cattle. AB - BACKGROUND: Natural heterologous valved conduits with a diameter greater than 22 mm that can be used for right ventricular outflow tract reconstruction in adults are not commercially available. The purpose of this study was to measure by ultrasonography the maximum diameter of the distended jugular veins of horses and cattle, respectively, to identify a population of animals that would be suitable for post-mortem collection of jugular veins at sizes greater than 22 mm. METHODS: The study population included 60 Warmblood horses, 25 Freiberger horses, 20 Brown Swiss cows, and 20 Holstein cows (including 10 Holstein and 10 Red Holstein). The maximum cross-sectional diameter of the distended jugular veins was measured at a location half-way between the mandibular angle and the thoracic inlet. The thoracic circumference (heart girth length) was used as a surrogate of body size. The jugular vein diameters of the different populations were compared by analysis of variance and the association between heart girth length and jugular vein diameter was determined in each of the four study populations by linear regression analysis. RESULTS: There was considerable individual variation of jugular vein diameters within each of the four study populations. There was no statistically significant relationship between thoracic circumference and jugular vein diameter in any of the populations. The jugular vein diameters of Brown Swiss cows were significantly larger than those of any of the other populations. Warmblood horses had significantly larger jugular vein diameters compared to Freiberger horses. CONCLUSION: The results of this study suggest that the production of bovine or equine xenografts with diameters of greater than 22 mm would be feasible. Differences between species and breeds need to be considered. However, prediction of the jugular vein diameter based on breed and heart girth length in an individual animal is inaccurate. PMID- 19678941 TI - Refractoriness of hepatitis C virus internal ribosome entry site to processing by Dicer in vivo. AB - BACKGROUND: Hepatitis C virus (HCV) is a positive-strand RNA virus harboring a highly structured internal ribosome entry site (IRES) in the 5' nontranslated region of its genome. Important for initiating translation of viral RNAs into proteins, the HCV IRES is composed of RNA structures reminiscent of microRNA precursors that may be targeted by the host RNA silencing machinery. RESULTS: We report that HCV IRES can be recognized and processed into small RNAs by the human ribonuclease Dicer in vitro. Furthermore, we identify domains II, III and VI of HCV IRES as potential substrates for Dicer in vitro. However, maintenance of the functional integrity of the HCV IRES in response to Dicer overexpression suggests that the structure of the HCV IRES abrogates its processing by Dicer in vivo. CONCLUSION: Our results suggest that the HCV IRES may have evolved to adopt a structure or a cellular context that is refractory to Dicer processing, which may contribute to viral escape of the host RNA silencing machinery. PMID- 19678942 TI - Xenograft models of head and neck cancers. AB - Head and neck cancers are among the most prevalent tumors in the world. Despite advances in the treatment of head and neck tumors, the survival of patients with these cancers has not markedly improved over the past several decades because of our inability to control and our poor understanding of the regional and distant spread of this disease. One of the factors contributing to our poor understanding may be the lack of reliable animal models of head and neck cancer metastasis. The earliest xenograft models in which human tumor cells were grown in immunosuppressed mice involved subcutaneous implantation of human head and neck cancer cell lines. Subcutaneous xenograft models have been popular because they are easy to establish, easy to manage, and lend themselves to ready quantitation of the tumor burden. More recently, orthotopic xenograft models, in which the tumor cells are implanted in the tumor site of origin, have been used with greater frequency in animal studies of head and neck cancers. Orthotopic xenograft models are advantageous for their ability to mimic local tumor growth and recapitulate the pathways of metastasis seen in human head and neck cancers. In addition, recent innovations in cell labeling techniques and small-animal imaging have enabled investigators to monitor the metastatic process and quantitate the growth and spread of orthopically implanted tumors. This review summarizes the progress in the development of murine xenograft models of head and neck cancers. We then discuss the advantages and disadvantages of each type of xenograft model. We also discuss the potential for these models to help elucidate the mechanisms of regional and distant metastasis, which could improve our ability to treat head and neck cancers. PMID- 19678943 TI - Blood transfusion in the critically ill: does storage age matter? AB - Morphologic and biochemical changes occur during red cell storage prior to product expiry, and these changes may hinder erythrocyte viability and function following transfusion. Despite a relatively large body of literature detailing the metabolic and structural deterioration that occurs during red cell storage, evidence for a significant detrimental clinical effect related to the transfusion of older blood is relatively less conclusive, limited primarily to observations in retrospective studies. Nonetheless, the implication that the transfusion of old, but not outdated blood may have negative clinical consequences demands attention. In this report, the current understanding of the biochemical and structural changes that occur during storage, known collectively as the storage lesion, is described, and the clinical evidence concerning the detrimental consequences associated with the transfusion of relatively older red cells is critically reviewed. Although the growing body of literature demonstrating the deleterious effects of relatively old blood is compelling, it is notable that all of these reports have been retrospective, and most of these studies have evaluated patients who received a mixture of red cell units of varying storage age. Until prospective studies have been completed and produce confirmative results, it would be premature to recommend any modification of current transfusion practice regarding storage age. In 1917, Frances Payton Rous and J.R. Turner identified that a citrate-glucose solution allowed for the preservation of a whole blood unit for up to five days, thus facilitating the formative practice of blood banking. Later, Loutit and Mollison of Great Britain developed the first anticoagulant of the modern era, known as acid-citrate-dextrose (ACD). ACD extended the shelf life of refrigerated blood to 21 days, and ACD remained in wide spread usage until the 1960s, when it was replaced by citrate-phosphate dextrose (CPD) and citrate-phosphate-dextrose-adenine (CPDA) solutions that increased shelf life to 35 days and 42 days respectively. More recently, additive solutions containing saline, adenine, and dextrose have been developed to augment red cell survival following transfusion, although without any direct increase in storage duration. It is now well appreciated, however, that a number of morphologic and biochemical changes occur during red cell storage prior to product expiry, and these changes may hinder erythrocyte viability and function following transfusion. Despite a relatively large body of literature detailing the metabolic and structural deterioration that occurs during red cell storage, evidence for a significant detrimental clinical effect related to the transfusion of older blood is relatively less conclusive, limited primarily to observations in retrospective studies. Nonetheless, the implication that the transfusion of old, but not outdated blood may have negative clinical consequences demands attention. The purpose of this report is to describe the current understanding of the biochemical and structural changes that occur during storage, known collectively as the storage lesion, and to critically review the clinical evidence concerning the detrimental consequences associated with the transfusion of relatively older red cells. PMID- 19678944 TI - Primary care-based multifaceted, interdisciplinary medical educational intervention for patients with systolic heart failure: lessons learned from a cluster randomised controlled trial. AB - BACKGROUND: Chronic (systolic) heart failure (CHF) is a common and disabling condition. Adherence to evidence-based guidelines in primary care has been shown to improve health outcomes. The aim was to explore the impact of a multidisciplinary educational intervention for general practitioners (GPs) (Train the trainer = TTT) on patient and performance outcomes. METHODS: This paper presents the key findings from the trial and discusses the lessons learned during the implementation of the TTT trial. Primary care practices were randomly assigned to the TTT intervention or to the control group. 37 GPs (18 TTT, 19 control) were randomised and 168 patients diagnosed with ascertained CHF (91 TTT, 77 control) were enrolled. GPs in the intervention group attended four meetings addressing clinical practice guidelines and pharmacotherapy feedback. The primary outcome was patient self-reported quality of life at seven months, using the SF 36 Physical Functioning scale. Secondary outcomes included other SF-36 scales, the Kansas City Cardiomyopathy Questionnaire (KCCQ), total mortality, heart failure hospital admissions, prescribing, depressive disorders (PHQ-9), behavioural change (European Heart Failure Self-Care Behaviour Scale), patient perceived quality of care (EUROPEP) and improvement of heart failure using NT proBNP-levels. Because recruitment targets were not achieved an exploratory analysis was conducted. RESULTS: There was high baseline achievement in both groups for many outcomes. At seven months, there were no significant mean difference between groups for the primary outcome measure (-3.3, 95%CI -9.7 to 3.1, p = 0.30). The only difference in secondary outcomes related to the prescribing of aldosterone antagonists by GPs in the intervention group, with significant between group differences at follow-up (42 vs. 24%, adjusted OR = 4.0, 95%CI 1.2-13; p = 0.02). CONCLUSION: The intervention did not change the primary outcome or most secondary outcomes. Recruitment targets were not achieved and the under-recruitment of practices and patients alongside a selection bias of participating GPs, prohibit definite conclusions, but the CI indicates a non effectiveness of the intervention in this sample. We describe the lessons learned from conducting the trial for the future planning and conduct of confirmatory trials in primary care. TRIAL REGISTRATION: ISRCTN08601529. PMID- 19678945 TI - Loss of matK RNA editing in seed plant chloroplasts. AB - BACKGROUND: RNA editing in chloroplasts of angiosperms proceeds by C-to-U conversions at specific sites. Nuclear-encoded factors are required for the recognition of cis-elements located immediately upstream of editing sites. The ensemble of editing sites in a chloroplast genome differs widely between species, and editing sites are thought to evolve rapidly. However, large-scale analyses of the evolution of individual editing sites have not yet been undertaken. RESULTS: Here, we analyzed the evolution of two chloroplast editing sites, matK-2 and matK 3, for which DNA sequences from thousands of angiosperm species are available. Both sites are found in most major taxa, including deep-branching families such as the nymphaeaceae. However, 36 isolated taxa scattered across the entire tree lack a C at one of the two matK editing sites. Tests of several exemplary species from this in silico analysis of matK processing unexpectedly revealed that one of the two sites remain unedited in almost half of all species examined. A comparison of sequences between editors and non-editors showed that specific nucleotides co-evolve with the C at the matK editing sites, suggesting that these nucleotides are critical for editing-site recognition. CONCLUSION: (i) Both matK editing sites were present in the common ancestor of all angiosperms and have been independently lost multiple times during angiosperm evolution.(ii) The editing activities corresponding to matK-2 and matK-3 are unstable.(iii) A small number of third-codon positions in the vicinity of editing sites are selectively constrained independent of the presence of the editing site, most likely because of interacting RNA-binding proteins. PMID- 19678946 TI - Preferences for treatment of Attention-Deficit/Hyperactivity Disorder (ADHD): a discrete choice experiment. AB - BACKGROUND: While there is an increasing emphasis on patient empowerment and shared decision-making, subjective values for attributes associated with their treatment still need to be measured and considered. This contribution seeks to define properties of an ideal drug treatment of individuals concerned with Attention-Deficit/Hyperactivity Disorder (ADHD). Because of the lack of information on patient needs in the decision-makers assessment of health services, the individuals' preferences often play a subordinate role at present. Discrete Choice Experiments offer strategies for eliciting subjective values and making them accessible for physicians and other health care professionals. METHODS: The evidence comes from a Discrete Choice Experiments (DCE) performed in 2007. After reviewing the literature about preferences of ADHS we conducted a qualitative study with four focus groups consisting of five to eleven ADHS patients each. In order to achieve content validity, we aimed at collecting all relevant factors for an ideal ADHS treatment. In a subsequent quantitative study phase (n = 219), data was collected in an online or paper-pencil self-completed questionnaire. It included sociodemographic data, health status and patients' preferences of therapy characteristics using direct measurement (23 items on a five-point Likert-scale) as well as a Discrete-Choice-Experiment (DCE, six factors in a fold-over design). RESULTS: Those concerned were capable of clearly defining success criteria and expectations. In the direct assessment and the DCE, respondents attached special significance to the improvement of their social situation and emotional state (relative importance 40%). Another essential factor was the desire for drugs with a long-lasting effect over the day (relative importance 18%). Other criteria, such as flexibility and discretion, were less important to the respondents (6% and 9%, respectively). CONCLUSION: Results point out that ADHD patients and their family members have clear ideas of their needs. This is especially important against the backdrop of present discussions in the healthcare sector on the relevance of patient reported outcomes (PROs) and shared decision-making. The combination of the methods used in this study offer promising strategies to elicit subjective values and making them accessible for health care professionals in a manner that drives health choices. PMID- 19678947 TI - Associations among parental feeding styles and children's food intake in families with limited incomes. AB - BACKGROUND: Although general parenting styles and restrictive parental feeding practices have been associated with children's weight status, few studies have examined the association between feeding styles and proximal outcomes such as children's food intake, especially in multi-ethnic families with limited incomes. The purpose of this study was to evaluate the association of parental feeding styles and young children's evening food intake in a multiethnic sample of families in Head Start. METHODS: Participants were 715 Head Start children and their parents from Texas and Alabama representing three ethnic groups: African American (43%), Hispanic (29%), and White (28%). The Caregivers Feeding Styles Questionnaire (Hughes) was used to characterize authoritative, authoritarian (referent), indulgent or uninvolved feeding styles. Food intake in several food groups was calculated from 3 days of dietary recalls for the child for evening food intakes from 3 PM until bedtime. RESULTS: Compared to children of authoritarian parents, intakes of fruits, juice and vegetables were lowest among children of indulgent or uninvolved parents (1.77 +/- 0.09 vs 1.45 +/- 0.09 and 1.42 +/- 0.11 cups) as were intakes of dairy foods (0.84 +/- 0.05 vs 0.67 +/- 0.05 and 0.63+0.06 cups), respectively. CONCLUSION: Findings suggest that permissive parent feeding styles like indulgent or uninvolved relate negatively to children's intake of nutrient-rich foods fruit, 100% fruit juice, vegetables and dairy foods from 3 PM until bedtime. PMID- 19678948 TI - Skeletal muscle structural lipids improve during weight-maintenance after a very low calorie dietary intervention. AB - BACKGROUND: The objective was to investigate in a group of obese subjects the course in skeletal muscle phospholipid (SMPL) fatty acids (FA) during a 24-weeks weight maintenance program, which was preceded by a successful very low calorie dietary intervention (VLCD). Special focus was addressed to SMPL omega-3 FA, which is a lipid entity that influences insulin action. METHODS: Nine obese subjects (BMI = 35.7 +/- 1.0 kg/m(2)), who had completed an 8 weeks VLCD (weight loss = -9.7 +/- 1.6 kg, P < 0.001), had obtained skeletal muscle biopsies (vastus lateralis) before and after a dietician-guided 24-weeks weight-maintenance program (-1.2 +/- 1.5 kg, P = ns). SMPL FA composition was determined by gas liquid chromatography. During the preceding VLCD, insulin sensitivity (HOMA-IR) and glycemic control (HbA1c) improved but no change in SMPL omega-3 FA was observed. During the weight-maintenance program five subjects received the pancreas lipase inhibitor Orlistat 120 mg t.i.d. versus placebo. RESULTS: HOMA-IR and HbA1c stabilized and SMPL total omega-3 FA, docosahexaenoic acid and ratio of n-3/n-6 polyunsaturated FA increased by 24% (P < 0.01), 35% (P < 0.02) and 26% (P < 0.01), respectively, whereas saturated and monounsaturated FA did not change. Plasma total-cholesterol and LDL-cholesterol, which decreased during the VLCD, reverted to pre-VLCD levels (P < 0.01). Orlistat therapy was associated with weight-loss (P < 0.05), trends for better glycemic control (P = 0.15) and greater increase in SMPL docosahexaenoic acid (P = 0.12) but similar reversal of plasma cholesterols compared to placebo. CONCLUSION: The data are consistent with the notion that greater SMPL omega-3 FA obtained during a weight-maintenance program may play a role for preserving insulin sensitivity and glycemic control being generated during a preceding VLCD. PMID- 19678949 TI - Application of the indirect fluorescent antibody assay in the study of malaria infection in the Yangtze River Three Gorges Reservoir, China. AB - BACKGROUND: China Yangtze Three Gorges Project (TGP) is one of the biggest construction projects in the world. The areas around the Three Gorge Dam has a history of tertian malaria and subtertian malaria epidemic, but there are no overall data about malaria epidemics before the completion of the project. The objective of this study was to get a reliable baseline on malaria infection in the Yangtze River Three Gorges reservoir area and to provide reference data for future studies about the impact of the project on malaria epidemics. METHODS: Two surveys of malaria infection were carried out in area, at six-month intervals in May and October 2008. About 3,600 dual specimens blood film samples for parasite diagnosis and filter paper blood spots for serology (using the immunofluorescence antibody test) were collected from the general population, including school populations, whenever possible. RESULTS: The overall percentage of positive response of the same population during post-transmission periods was about twice (1.40/0.72) of that in pre-transmission. Positive individuals under 15 years of age were detected in all the localities. CONCLUSION: A certain extent of malaria infection existed in this area. Additional studies are needed to determine the length of malaria experience, and chemotherapeutic intervention as well as the distribution of main vectors for transmission in this area. PMID- 19678950 TI - Healing relationships and the existential philosophy of Martin Buber. AB - The dominant unspoken philosophical basis of medical care in the United States is a form of Cartesian reductionism that views the body as a machine and medical professionals as technicians whose job is to repair that machine. The purpose of this paper is to advocate for an alternative philosophy of medicine based on the concept of healing relationships between clinicians and patients. This is accomplished first by exploring the ethical and philosophical work of Pellegrino and Thomasma and then by connecting Martin Buber's philosophical work on the nature of relationships to an empirically derived model of the medical healing relationship. The Healing Relationship Model was developed by the authors through qualitative analysis of interviews of physicians and patients. Clinician-patient healing relationships are a special form of what Buber calls I-Thou relationships, characterized by dialog and mutuality, but a mutuality limited by the inherent asymmetry of the clinician-patient relationship. The Healing Relationship Model identifies three processes necessary for such relationships to develop and be sustained: Valuing, Appreciating Power and Abiding. We explore in detail how these processes, as well as other components of the model resonate with Buber's concepts of I-Thou and I-It relationships. The resulting combined conceptual model illuminates the wholeness underlying the dual roles of clinicians as healers and providers of technical biomedicine. On the basis of our analysis, we argue that health care should be focused on healing, with I-Thou relationships at its core. PMID- 19678951 TI - Development of a RVFV ELISA that can distinguish infected from vaccinated animals. AB - BACKGROUND: Rift Valley Fever Virus is a pathogen of humans and livestock that causes significant morbidity and mortality throughout Africa and the Middle East. A vaccine that would protect animals from disease would be very beneficial to the human population because prevention of the amplification cycle in livestock would greatly reduce the risk of human infection by preventing livestock epizootics. A mutant virus, constructed through the use of reverse genetics, is protective in laboratory animal models and thus shows promise as a potential vaccine. However, the ability to distinguish infected from vaccinated animals is important for vaccine acceptance by national and international authorities, given regulations restricting movement and export of infected animals. RESULTS: In this study, we describe the development of a simple assay that can be used to distinguish naturally infected animals from ones that have been vaccinated with a mutant virus. We describe the cloning, expression and purification of two viral proteins, and the development of side by side ELISAs using the two viral proteins. CONCLUSION: A side by side ELISA can be used to differentiate infected from vaccinated animals. This assay can be done without the use of biocontainment facilities and has potential for use in both human and animal populations. PMID- 19678952 TI - Exploring the optimum approach to the use of CT densitometry in a randomised placebo-controlled study of augmentation therapy in alpha 1-antitrypsin deficiency. AB - BACKGROUND: Computed tomography (CT) lung densitometry has been demonstrated to be the most sensitive and specific outcome measure for the assessment of emphysema-modifying therapy, but the optimum densitometric index has yet to be determined and targeted sampling may be more sensitive than whole lung assessment. The EXAcerbations and CT scan as Lung Endpoints (EXACTLE) trial aimed to clarify the optimum approach to the use of CT densitometry data for the assessment of alpha 1-antitrypsin (AAT) augmentation therapy on the progression of emphysema in AAT deficiency (AATD). METHODS: Patients with AATD (n = 77) were randomised to weekly infusions of 60 mg/kg human AAT (Prolastin) or placebo over 2 to 2.5 years. Lung volume was included as a covariate in an endpoint analysis and a comparison was made of different CT densitometric indices (15th percentile lung density [PD15], mean lung density [MLD] and voxel index at a threshold of 910 [VI-910] and -950 [VI-950] Hounsfield Units) obtained from whole lung scans at baseline and at 24 to 30 months. Targeted regional sampling was compared with whole lung assessment. RESULTS: Whole lung analysis of the total change (baseline to last CT scan) compared with placebo indicated a concordant trend that was suggestive of a treatment effect for all densitometric indices (MLD [1.402 g/L, p = 0.204]; VI-910 [-0.611, p = 0.389]; VI-950 [-0.432, p = 0.452]) and that was significant using PD15 (1.472 g/L, p = 0.049). Assessment of the progression of emphysema in the apical, middle and basal regions of the lung by measurement with PD15 showed that this treatment effect was more evident when the basal third was sampled (1.722 g/L, p = 0.040). A comparison between different densitometric indices indicated that the influence of inspiratory variability between scans was greatest for PD15, but when adjustment for lung volume was made this index was the most sensitive measure of emphysema progression. CONCLUSION: PD15 is the most sensitive index of emphysema progression and of treatment modification. Targeted sampling may be more sensitive than whole lung analysis. TRIAL REGISTRATION: Registered in ClinicalTrials.gov as 'Antitrypsin (AAT) to Treat Emphysema in AAT Deficient Patients'; ClinicalTrials.gov Identifier: NCT00263887. PMID- 19678953 TI - Archaeosomes made of Halorubrum tebenquichense total polar lipids: a new source of adjuvancy. AB - BACKGROUND: Archaeosomes (ARC), vesicles prepared from total polar lipids (TPL) extracted from selected genera and species from the Archaea domain, elicit both antibody and cell-mediated immunity to the entrapped antigen, as well as efficient cross priming of exogenous antigens, evoking a profound memory response. Screening for unexplored Archaea genus as new sources of adjuvancy, here we report the presence of two new Halorubrum tebenquichense strains isolated from grey crystals (GC) and black mood (BM) strata from a littoral Argentinean Patagonia salt flat. Cytotoxicity, intracellular transit and immune response induced by two subcutaneous (sc) administrations (days 0 and 21) with BSA entrapped in ARC made of TPL either form BM (ARC-BM) and from GC (ARC-GC) at 2% w/w (BSA/lipids), to C3H/HeN mice (25 microg BSA, 1.3 mg of archaeal lipids per mouse) and boosted on day 180 with 25 microg of bare BSA, were determined. RESULTS: DNA G+C content (59.5 and 61.7% mol BM and GC, respectively), 16S rDNA sequentiation, DNA-DNA hybridization, arbitrarily primed fingerprint assay and biochemical data confirmed that BM and GC isolates were two non-previously described strains of H. tebenquichense. Both multilamellar ARC mean size were 564 +/- 22 nm, with -50 mV zeta-potential, and were not cytotoxic on Vero cells up to 1 mg/ml and up to 0.1 mg/ml of lipids on J-774 macrophages (XTT method). ARC inner aqueous content remained inside the phago-lysosomal system of J-774 cells beyond the first incubation hour at 37 degrees C, as revealed by pyranine loaded in ARC. Upon subcutaneous immunization of C3H/HeN mice, BSA entrapped in ARC-BM or ARC-GC elicited a strong and sustained primary antibody response, as well as improved specific humoral immunity after boosting with the bare antigen. Both IgG1 and IgG2a enhanced antibody titers could be demonstrated in long-term (200 days) recall suggesting induction of a mixed Th1/Th2 response. CONCLUSION: We herein report the finding of new H. tebenquichense non alkaliphilic strains in Argentinean Patagonia together with the adjuvant properties of ARC after sc administration in mice. Our results indicate that archaeosomes prepared with TPL from these two strains could be successfully used as vaccine delivery vehicles. PMID- 19678954 TI - Yeast artificial chromosomes employed for random assembly of biosynthetic pathways and production of diverse compounds in Saccharomyces cerevisiae. AB - BACKGROUND: Natural products are an important source of drugs and other commercially interesting compounds, however their isolation and production is often difficult. Metabolic engineering, mainly in bacteria and yeast, has sought to circumvent some of the associated problems but also this approach is impeded by technical limitations. Here we describe a novel strategy for production of diverse natural products, comprising the expression of an unprecedented large number of biosynthetic genes in a heterologous host. RESULTS: As an example, genes from different sources, representing enzymes of a seven step flavonoid pathway, were individually cloned into yeast expression cassettes, which were then randomly combined on Yeast Artificial Chromosomes and used, in a single transformation of yeast, to create a variety of flavonoid producing pathways. Randomly picked clones were analysed, and approximately half of them showed production of the flavanone naringenin, and a third of them produced the flavonol kaempferol in various amounts. This reflected the assembly of 5-7 step multi species pathways converting the yeast metabolites phenylalanine and/or tyrosine into flavonoids, normally only produced by plants. Other flavonoids were also produced that were either direct intermediates or derivatives thereof. Feeding natural and unnatural, halogenated precursors to these recombinant clones demonstrated the potential to further diversify the type of molecules that can be produced with this technology. CONCLUSION: The technology has many potential uses but is particularly suited for generating high numbers of structurally diverse compounds, some of which may not be amenable to chemical synthesis, thus greatly facilitating access to a huge chemical space in the search for new commercially interesting compounds. PMID- 19678955 TI - Aerobic exercise and intraocular pressure in normotensive and glaucoma patients. AB - BACKGROUND: With the increasing number of people participating in physical aerobic exercise, jogging in particular, we considered that it would be worth knowing if there are should be limits to the exercise with regard to the intraocular pressure (IOP) of the eyes. The purpose of this study is to check IOP in healthy and primary glaucoma patients after aerobic exercise. METHODS: 145 individuals were subdivided into seven groups: normotensives who exercised regularly (Group A); normotensives in whose right eye (RE) timolol maleate 0.5% (Group B), latanoprost 0.005% (Group C), or brimonidine tartrate 0.2% (Group D) was instilled; and primary glaucoma patients under monotherapy with beta-blockers (Group E), prostaglandin analogues (Group F) or combined antiglaucoma treatment (Group G) instilled in both eyes. The IOP of both eyes was measured before and after exercise. RESULTS: A statistically significant decrease was found in IOP during jogging. The aerobic exercise reduces the IOP in those eyes where a b blocker, a prostaglandin analogue or an alpha-agonist was previously instilled. The IOP is also decreased in glaucoma patients who are already under antiglaucoma treatment. CONCLUSION: There is no ocular restriction for simple glaucoma patients in performing aerobic physical activity. PMID- 19678956 TI - Anti-inflammatory and anti-coagulatory activities of caffeic acid and ellagic acid in cardiac tissue of diabetic mice. AB - BACKGROUND: Caffeic acid (CA) and ellagic acid (EA) are phenolic acids naturally occurring in many plant foods. Cardiac protective effects of these compounds against dyslipidemia, hypercoagulability, oxidative stress and inflammation in diabetic mice were examined. METHODS: Diabetic mice were divided into three groups (15 mice per group): diabetic mice with normal diet, 2% CA treatment, or 2% EA treatment. One group of non-diabetic mice with normal diet was used for comparison. After 12 weeks supplement, mice were sacrificed, and the variation of biomarkers for hypercoagulability, oxidative stress and inflammation in cardiac tissue of diabetic mice were measured. RESULTS: The intake of CA or EA significantly increased cardiac content of these compounds, alleviated body weight loss, elevated plasma insulin and decreased plasma glucose levels in diabetic mice (p < 0.05). These treatments also significantly enhanced plasma antithrombin-III and protein C activities (p < 0.05); and decreased triglyceride content in cardiac tissue and plasma (p < 0.05), in which the hypolipidemic effects of EA were significantly greater than that of CA (p < 0.05). CA or EA significantly lowered cardiac levels of malondialdehyde, reactive oxygen species, interleukin (IL)-beta, IL-6, tumor necrosis factor (TNF)-alpha and monocyte chemoattractant protein (MCP)-1 (p < 0.05); and retained cardiac activity of glutathione peroxidase (GPX), superoxide dismutase (SOD) and catalase (p < 0.05). These compounds also significantly up-regulated cardiac mRNA expression of GPX1, SOD and catalase; and down-regulated IL-1beta, IL-6, TNF-alpha and MCP-1 mRNA expression in diabetic mice (p < 0.05). CONCLUSION: These results support that CA and EA could provide triglyceride-lowering, anti-coagulatory, anti-oxidative, and anti-inflammatory protection in cardiac tissue of diabetic mice. Thus, the supplement of these agents might be helpful for the prevention or attenuation of diabetic cardiomyopathy. PMID- 19678957 TI - The distribution and transitions of physicians in Japan: a 1974-2004 retrospective cohort study. AB - BACKGROUND: In Japan, physicians freely choose their specialty and workplace, because to date there is no management system to ensure a balanced distribution of physicians. Physicians in Japan start their careers in hospitals, then become specialists, and then gradually leave hospitals to work in private clinics and take on primary care roles in their specialty fields. The present study aimed to analyse national trends in the distribution and career transitions of physicians among types of facilities and specialties over a 30-year period. METHODS: We obtained an electronic file containing physician registration data from the Survey of Physicians, Dentists and Pharmacists. Descriptive statistics and data on movement between facilities (hospitals and clinics) for all physicians from 1974, 1984, 1994 and 2004 were analysed. Descriptive statistics for the groups of physicians who graduated in 1970, 1980 and 1990 were also analysed, and we examined these groups over time to evaluate their changes of occupation and specialty. RESULTS: The number of physicians per 100,000 population was 113 in 1974, and rose to 212 by 2004. The number of physicians working in hospitals increased more than threefold. In Japan, while almost all physicians choose hospital-based positions at the beginning of their career, around 20% of physicians withdrew from hospitals within 10 years, and this trend of leaving hospitals was similar among generations. Physicians who graduated in 1980 and registered in general surgery, cardiovascular surgery or paediatric surgery were 10 times more likely to change their specialty, compared with those who registered in internal medicine. More than half of the physicians who registered in 1970 had changed their specialties within a period of 30 years. CONCLUSION: The government should focus primarily on changing the physician fee schedule, with careful consideration of the balance between office-based physicians and hospital-based physicians and among specialties. To implement effective policies in managing health care human resources, policy-makers should also pay attention to continuously monitoring physicians' practising status and career motivations; and national consensus is needed regarding the number of physicians required in each type of facility and specialty as well as region. PMID- 19678958 TI - Ethical issues relating the the banking of umbilical cord blood in Mexico. AB - BACKGROUND: Umbilical cord banks are a central component, as umbilical cord tissue providers, in both medical treatment and scientific research with stem cells. But, whereas the creation of umbilical cord banks is seen as successful practice, it is perceived as a risky style of play by others. This article examines and discusses the ethical, medical and legal considerations that arise from the operation of umbilical cord banks in Mexico. DISCUSSION: A number of experts have stated that the use of umbilical cord goes beyond the mere utilization of human tissues for the purpose of treatment. This tissue is also used in research studies: genetic studies, studies to evaluate the effectiveness of new antibiotics, studies to identify new proteins, etc. Meanwhile, others claim that the law and other norms for the functioning of cord banks are not consistent and are poorly defined. Some of these critics point out that the confidentiality of donor information is handled differently in different places. The fact that private cord banks offer their services as "biological insurance" in order to obtain informed consent by promising the parents that the tissue that will be stored insures the health of their child in the future raises the issue of whether the consent is freely given or given under coercion. Another consideration that must be made in relation to privately owned cord banks has to do with the ownership of the stored umbilical cord. SUMMARY: Conflicts between moral principles and economic interests (non-moral principles) cause dilemmas in the clinical practice of umbilical cord blood storage and use especially in privately owned banks. This article presents a reflection and some of the guidelines that must be followed by umbilical cord banks in order to deal with these conflicts. This reflection is based on the fundamental notions of ethics and public health and seeks to be a contribution towards the improvement of umbilical cord banks' performance. PMID- 19678959 TI - Incidence of pelvic inflammatory disease in a large cohort of women tested for Chlamydia trachomatis: a historical follow-up study. AB - BACKGROUND: Chlamydia trachomatis is a highly prevalent sexually transmitted disease. Testing rates among young Norwegian women are high. Young women diagnosed with C. trachomatis are often worried about future complications. METHODS: Our cohort consisted of 24,947 women born 1970-1984 who were tested for C. trachomatis infection during 1990-2005. We linked C. trachomatis laboratory data to data on hospitalizations for pelvic inflammatory disease during 1990 2005. Cox regression analysis with time-dependent covariates adjusted for age at first test was used to assess the association between C. trachomatis history and pelvic inflammatory disease. RESULTS: Follow-up until the end of 2005 included 201,387 woman-years. The incidence rate of hospitalization for pelvic inflammatory disease was higher among women with prior C. trachomatis infection than among women with negative tests only (48 events during 32,057 person-years and 143 events during 169,192 person-years, corresponding to 0.15 and 0.08 per 100 person-years, respectively). The corresponding hazard ratio adjusted for age at first test was 1.69 (95% CI, 1.21-2.36). CONCLUSION: Our data show a link between a diagnosis of C. trachomatis infection and subsequent pelvic inflammatory disease. However, pelvic inflammatory disease was a rare event irrespective of C. trachomatis status. These, together with other recent findings, can be used to reassure women worried about their future reproductive health following a diagnosis of C. trachomatis. PMID- 19678960 TI - Interactions of socioeconomic position with psychosocial and environmental correlates of children's physical activity: an observational study of South Australian families. AB - BACKGROUND: Evidence for psychosocial and environmental correlates on children's physical activity is scattered and somewhat unconvincing. Further, the moderating influences of socioeconomic position (SEP) on these influences are largely unexplored. The aim of this study was to examine the interactions of SEP, operationalised by mother education, and predictors of children's physical activity based on the Youth Physical Activity Promotion Model. METHODS: In 2005, a sample of South Australians (10-15 y) was surveyed on psychosocial and environmental correlates of physical activity using the Children's Physical Activity Correlates Questionnaire (n = 3300) and a parent survey (n = 1720). The following constructs were derived: 'is it worth it?' (perceived outcomes); 'am I able?' (perceived competency); 'reinforcing' (parental support); and 'enabling' (parent-perceived barriers). Self-reported physical activity was represented by a global score derived from the Physical Activity Questionnaire for Adolescents. Associations among physical activity and hypothesised correlates were tested among children with mothers of high (university educated) and low (left school at or before 15 y) SEP. RESULTS: Among high SEP children, 'is it worth it?' emerged as a significant predictor of physical activity for boys and girls. Among low SEP children, 'is it worth it?' predicted boys' physical activity, while among girls, 'reinforcing' was the only significant predictor, explaining ~35% of the total explained variance in physical activity. CONCLUSION: While perceived outcomes emerged as a consistent predictor of physical activity in this sample, parental support was a powerful limiting factor among low SEP girls. Interventions among this high risk group should focus on supporting parents to provide both emotional and instrumental support for their daughters to engage in physical activity. PMID- 19678961 TI - Target splitting in radiation therapy for lung cancer: further developments and exemplary treatment plans. AB - BACKGROUND: Reporting further developments evolved since the first report about this conformal technique. METHODS: Technical progress focused on optimization of the quality assurance (QA) program, especially regarding the required work input; and on optimization of beam arrangements. RESULTS: Besides performing the regular QA program, additional time consuming dosimetric measurements and verifications no longer have to be accomplished.'Class solutions' of treatment plans for six patients with non-resected non-small cell lung cancer in locally advanced stages are presented. Target configurations comprise one central and five peripheral tumor sites with different topographic positions to hilus and mediastinum. The mean dose to the primary tumor is 81,9 Gy (range 79,2-90,0 Gy), to macroscopically involved nodes 61,2 Gy (range 55,8-63,0 Gy), to electively treated nodes 45,0 Gy. Treatments are performed twice daily, with fractional doses of 1,8 Gy at an interval of 11 hours. Median overall treatment time is 33 days. The set-up time at the linac does not exceed the average time for any other patient. CONCLUSION: Target splitting is a highly conformal and nonetheless non expensive method with regard to linac and staff time. It enables secure accelerated high-dose treatments of patients with NSCLC. PMID- 19678962 TI - Alpha adrenergic modulation on effects of norepinephrine transporter inhibitor reboxetine in five-choice serial reaction time task. AB - The study examined the effects of a norepinephrine transporter (NET) inhibitor reboxetine (RBX) on an attentional performance test. Adult SD rats trained with five-choice serial reaction time task (5-CSRTT) were administered with RBX (0, 3.0 and 10 mg/kg) in the testing day. Alpha-1 adrenergic receptor antagonist PRA and alpha-2 adrenergic receptor antagonist RX821002 were used to clarify the RBX effect. Results revealed that rat received RBX at 10 mg/kg had an increase in the percentage of the correct response and decreases in the numbers of premature response. Alpha-1 adrenergic receptor antagonist Prazosin (PRA) at 0.1 mg/kg reversed the RBX augmented correct responding rate. However, alpha-2 adrenergic receptor antagonist RX821002 at 0.05 and 0.1 mg/kg dose dependently reversed the RBX reduced impulsive responding. Our results suggested that RBX as a norepinephrine transporter inhibitor can be beneficial in both attentional accuracy and response control and alpha-1 and alpha-2 adrenergic receptors might be involved differently. PMID- 19678963 TI - Genetic screening of 104 patients with congenitally malformed hearts revealed a fresh mutation of GATA4 in those with atrial septal defects. AB - We analysed the GATA binding protein 4 gene, or GATA4, along with the NK2 transcription factor related, locus 5 gene, or NKX2.5, to determine their genetic contribution to 104 sporadic patients in Indonesia with congenitally malformed hearts, 76 cases having atrial septal defect and 28 tetralogy of Fallot. We found only 1 novel mutation of GATA4 in those with atrial septal defects. Analysis of the genetic background of the parents of the patient showed for the first time that a new mutation of GATA4 can cause sporadic atrial septal defects. We failed to discover any other mutations of either the GATA4 or NKX2-5 genes, supporting the marked genetic heterogeneity of human congenital cardiac defects. PMID- 19678965 TI - Glutamatergic cerebellar granule neurons synthesize and secrete reelin in vitro. AB - In the postnatal forebrain, the extracellular matrix protein reelin is expressed and secreted by subsets of GABAergic neurons, whereas in the cerebellum reelin is detected in glutamatergic cells of the granule cell layer. Thus, various regions of the postnatal brain present different patterns of reelin expression, whose significance remains unknown. We combined immunocytochemical and pharmacological approaches to characterize the phenotypic and temporal profiles of reelin expression in dissociated cultures of cerebellar granule neurons. A single type of reelin immunoreactivity, identified by a punctate labelling, was present in the somata of the majority of neurons. This immunoreactivity was observed throughout maturation and was exclusively present in glutamatergic neurons expressing the vesicular glutamate transporter 1. Neurons containing the reelin receptors apolipoprotein E receptor 2 (Apoer2) and very low-density lipoprotein receptor (Vldlr) represented about 80% of cerebellar neurons. The vast majority of reelin-positive neurons coexpressed Apoer2, suggesting that reelin immunoreactivity resulted in part from receptor-bound reelin. Inhibition of protein synthesis with cycloheximide completely abolished reelin immunoreactivity. In contrast, blocking protein secretion with brefeldin A did not affect the proportion of punctate neurons but revealed a subpopulation of neurons characterized by a solid reelin staining. These data show for the first time that a homogeneous population of glutamatergic neurons can synthesize and secrete reelin in cerebellar granule cells in vitro. PMID- 19678966 TI - Adding cognitive behavioural treatment to either low-carbohydrate or low-fat diets: differential short-term effects. AB - To evaluate the efficacy of adding cognitive behavioural treatment (CBT) to either a low-carbohydrate (LC) diet or a low-fat (LF) diet in the treatment of weight loss of obese women, a randomised clinical intervention study was performed. A total of 105 healthy non-pregnant obese women (average age and BMI of 45.4 (sd 10.4) years and 36 (sd 4.3) kg/m2) were randomly allocated to the CBT or control (C) groups; within each group, women were randomly selected to receive either the LC or LF diet during 6 months. The pre-planned primary trial end-point was the weight loss. Differences between the groups were assessed using one-way ANOVA. There were three women (2.8 %) who dropped out, all of them in the CBT group. No differences in the anthropometric and laboratory characteristics at baseline were noted between women in the CBT (n 52) and control groups (n 50). Intention-to-treat analysis showed that weight loss in the CBT-LC (90 (sd 12.3) to 82.1 (sd 12.1) kg) and C-LC (89.4 (sd 10.0) to 85.8 (sd 9.8) kg) groups reached 8.7 and 4.0 %, respectively (P < 0.0001), and in the CBT-LF (87.9 (sd 11.4) to 79.4 (sd 11.8) kg) and C-LF (88.8 (sd 14.5) to 85.3 (sd 14.3) kg) groups it was 9.7 and 3.9 %, respectively (P < 0.05). Weight loss was higher in the CBT LF group than in the CBT-LC groups (P = 0.049). The present results showed that adding CBT to either the LF or LC diet produced significantly greater short-term weight loss in obese women compared with diet alone. These finding support the efficacy of CBT in breaking previous dietary patterns and in developing healthier attitudes that reinforce a healthier lifestyle. PMID- 19678967 TI - A prospective study of dietary intakes and influential factors from pregnancy to postpartum on maternal weight retention in Taipei, Taiwan. AB - Excessive gestational weight gain and postpartum weight retention are risk factors for female obesity. The present study was to examine dietary intakes and weight history from a prospective follow-up study from early pregnancy to 1 year postpartum. A total of 151 pregnant women within 20 weeks of pregnancy in Taipei, Taiwan were interviewed periodically to collect dietary and lifestyle information. The participants had an average age of 30 years and the average gestational weight gain was 14 kg, with an average daily intake of 7830 kJ (1870 kcal) in the 1 year following parturition. By bivariate analyses, maternal age, pre-pregnancy BMI and breast-feeding were not related to postpartum weight retention, but gestational weight gain had significant positive correlations (r 0.54 at 6 months, r 0.44 at 1 year; P < 0.05). The generalised estimating equations showed that the average weight before pregnancy, at 6 months and 1 year postpartum was 53.35 kg, 55.75 kg (weight retention 2.36 kg; P < 0.01) and 54.75 kg (weight retention 1.48 kg; P < 0.01), respectively. After controlling for age, pre-pregnancy BMI, gestational weight gain and parity, we found at 6 months that the adjusted weight retention at postpartum was 0.79 kg (P < 0.01), but at 1 year it was - 0.08 kg (P>0.05). From multivariate analyses, dietary energy intake and energy intake per kg body weight as a long-term physical activity index could explain 24 % of the variation at 6 months and 27 % of the variation at 1 year in postpartum weight retention. These results suggest that pregnant women should be advised to control gestational weight gain, decrease energy intakes after child bearing and maintain regular exercise in order to prevent postpartum obesity. PMID- 19678968 TI - Relationship between animal protein intake and muscle mass index in healthy women. AB - The amount and the type of dietary protein could play a role in determining the quantity of skeletal muscle mass. The aim was to examine the relationship between the type of protein intake and the level of muscle mass in healthy omnivorous and vegetarian Caucasian women. The design of the present study was an observational and cross-sectional study. Twenty-one omnivores (Om) and nineteen vegetarians (Ve) were recruited. Muscle mass index (urinary creatinine), dietary intake (5 d dietary records) and biochemical analyses (hormone, phyto-oestrogen and lipid profiles) were obtained. We found differences between groups for muscle mass (Ve: 18 kg v. Om: 23 kg; P = 0.010), muscle mass index (Ve: 6.7 kg/m2 v. Om: 8.3 kg/m2; P = 0.002), animal protein intake in g/d (P = 0.001) and in g/kg body weight per d (P = 0.003), plant protein intake in g/d (P = 0.015) and in g/kg body weight per d (P = 0.007), the animal:plant protein intake ratio (P = 0.001) and sex hormone-binding globulin (SHBG) (P = 0.001). Muscle mass index still correlated with animal protein intake in g/d (P = 0.001) and in g/kg body weight per d (P = 0.008), and the animal:plant protein intake ratio (P = 0.007) even after controlling for SHBG and plant protein intake. Finally, animal protein intake (g/d) was the independent predictor of muscle mass index (adjusted r2 0.42). Thus, a vegetarian diet is associated with a lower muscle mass index than is an omnivorous diet at the same protein intake. A good indicator of muscle mass index in women seems to be animal protein intake. PMID- 19678969 TI - BJN impact factor rises. PMID- 19678970 TI - The impact of sheep breed on the risk of classical scrapie. AB - The risk of classical scrapie in sheep is associated with polymorphisms in the prion protein (PrP) gene. In recent years, large-scale selective breeding programmes for sheep at lower risk of disease have been undertaken across the European Union. We analysed large-scale datasets on scrapie and sheep demography to investigate additional effects of sheep breed on scrapie risk. There was evidence for variation between certain breeds in the scrapie risk of some PrP genotypes, which could be caused by innate breed differences or distinct scrapie strains circulating within them. While the PrP genotypes of cases are generally consistent across breeds, some exceptions provide evidence that scrapie strain may influence affected PrP genotypes to a greater extent than innate breed differences. There was a significant association between the breed-level incidence of scrapie and the frequency of susceptible PrP genotypes in breeds. Our results lend support to selective breeding programmes which aim to reduce the frequency of high-risk PrP genotypes with measures not varying by sheep breed. PMID- 19678971 TI - El Tor cholera with severe disease: a new threat to Asia and beyond. AB - During epidemics of cholera in two rural sites (Bakerganj and Mathbaria), a much higher proportion of patients came for treatment with severe dehydration than was seen in previous years. V. cholerae O1 isolated from these patients was found to be El Tor in its phenotype, but its cholera toxin (CT) was determined to be that of classical biotype. Whether the observed higher proportion of severe dehydration produced by the El Tor biotype was due to a shift from El Tor to classical CT or due to other factors is not clear. However, if cholera due to strains with increased severity spread to other areas where treatment facilities are limited, there are likely to be many more cholera deaths. PMID- 19678972 TI - Effects of weather variability on infectious gastroenteritis. AB - Although multiple combinations of weather variability may contribute to an increased incidence of infectious gastrointestinal disease, few studies have investigated the association between weather variability and cases of infectious gastroenteritis. We acquired data for infectious gastroenteritis cases and weather variability in Fukuoka, Japan, from 1999 to 2007 and used time-series analysis to assess the effects of weather variability on infectious gastroenteritis cases, adjusting for confounding factors. In total, 422,176 infectious gastroenteritis cases were reported during the 9-year study period. The weekly number of infectious gastroenteritis cases increased by 7.7% (95% CI 4.6-10.8) for every 1 degrees C increase in the average temperature and by 2.3% (95% CI 1.4-3.1) for every 1% decrease in relative humidity. From 1999 to 2007, infectious gastroenteritis cases increased significantly with increased average temperature and decreased relative humidity in Fukuoka, Japan. PMID- 19678973 TI - Measles outbreak linked to a minority group in Austria, 2008. AB - We report on a measles outbreak originating in an anthroposophic community in Austria, 2008. A total of 394 (94.9%) cases fulfilled the outbreak case definition including 168 cases affiliated to the anthroposophic community. The source case was a school pupil from Switzerland. The Austrian outbreak strain was genotype D5, indistinguishable from the Swiss outbreak strain. A school-based retrospective cohort study in the anthroposophic school demonstrated a vaccine effectiveness of 97.3% in pupils who had received a single dose of measles containing vaccine and 100% in those who had received two doses. The vaccination coverage of the cases in the anthroposophic community was 0.6%. Of the 226 outbreak cases not belonging to the anthroposophic community, the 10-24 years age group was the most affected. Our findings underline the epidemiological significance of suboptimal vaccination coverage in anthroposophic communities and in older age groups of the general population in facilitating measles virus circulation. The findings of this outbreak investigation suggest that the WHO European Region is unlikely to achieve its 2010 target for measles and rubella elimination. PMID- 19678974 TI - Anaemia among pregnant Palestinian women in the Occupied Palestinian Territory. AB - OBJECTIVE: To assess anaemia prevalence and correlated social and biological determinants among pregnant women in the Occupied Palestinian Territory (oPt). DESIGN: A cross-sectional survey conducted among pregnant women attending/accessing UNRWA (United Nations Relief and Works Agency for Palestine Refugees in the Near East) health centres in the Gaza Strip and the West Bank in September and October 2006. SETTING: Fifty-five UNRWA health centres in the oPt (eighteen in the Gaza Strip and thirty-seven in the West Bank). SUBJECTS: A random sample of 1740 pregnant women. RESULTS: Overall anaemia prevalence was 38.6 % (95 % CI 36.3, 40.9 %). A substantial difference in anaemia prevalence was observed between the Gaza Strip and the West Bank (44.9 % v. 31.1 %, respectively), as well as a significant increase in anaemia prevalence in the Gaza Strip compared with an Agency-wide survey conducted in 2004 (44.9 % v. 35.7 %, respectively). Anaemia prevalence was found to increase with age, parity and trimester of gestation. CONCLUSIONS: Anaemia still appears to be a public health problem among pregnant women in spite of UNRWA interventions. The West Bank shows prevalence rates similar to those observed in neighbouring countries, while the Gaza Strip has higher rates. Prevalence rates of anaemia among pregnant Palestinian women are more than two times higher than those observed in Europe. PMID- 19678975 TI - The effect of the time interval between injection and parthenogenetic activation on the spindle formation and the in vitro developmental potential of somatic cell nuclear-transferred rat oocytes. AB - We examined the optimal conditions for somatic cell nuclear transfer (SCNT) in the rat. First, we examined the effect of preincubation time before activation on SCNT rat oocytes produced in the presence of MG132 with regard to spindle formation and the potential to develop into blastocysts. The spindles of SCNT oocytes continued to elongate with an increase in the culture duration and, in approximately half of oocytes, the chromosomes were distributed along the spindles at 120 min after incubation. Such abnormal spindle formation in SCNT oocytes is a possible reason for the low developmental potential of SCNT rat oocytes. To inhibit the formation of abnormal spindle formation, we examined secondly the developmental potential of rat SCNT oocytes that had been preincubated with nocodazole and demecolcine instead of MG132. The developmental rates in SCNT oocytes, however, were decreased. For successful rat somatic cell cloning, two steps might be required: (1) to culture the somatic cell nuclei for a sufficient time in MII oocyte cytoplasm to enhance nuclear reprogramming; and (2) to induce normal spindle formation with normal chromosomal construction. PMID- 19678976 TI - Double activation improves rabbit freeze-thawed oocytes developmental potential. AB - OBJECTIVE: To investigate the effects of various activation methods on freeze thawed rabbit oocytes developmental potential. METHODS: Rabbit oocytes were vitrified by cryoleafs and cryoprotected with ethylene glycol and propanediol. After thawing, the oocytes were fertilized by intracytoplasmic sperm injection (ICSI). Surviving oocytes after ICSI were divided into five groups at random. Group 1: Oocytes (n = 30) activated 1 h after ICSI by calcium ionomycin (I0634); Group 2: Oocytes (n = 26) activated by strontium chloride an hour after ICSI; Group 3: Oocytes (n = 33) activated by I0634 twice; Group 4: Oocytes (n = 28) were activated by strontium chloride twice; CONTROL GROUP: Inactivated oocytes (n = 39). Blastocysts derived from each group were transplanted to recipient rabbits. RESULTS: Rates of fertilization, cleavage and blastocyst formation of Group 3 were higher than those of Group 1 and Group 2 (81.8% vs 33.3% vs 53.8%, 54.5% vs 16.7% vs 26.9%, p < 0.05; 15.2% vs 3.3% vs 7.7%, p > 0.05). The rabbit transplanted with embryos derived from Group 3 became pregnant. Embryos derived from double activation could implant into endometrium. CONCLUSION: Double activation may increase freeze-thawed oocytes developmental potential. After activation, oocytes cleavage velocity may be faster than that of oocytes without activation. PMID- 19678981 TI - Influence of different metal matrices on manganese signal response in laser ablation inductively coupled plasma-mass spectrometry. AB - Laser ablation inductively coupled plasma-mass spectrometry (LA-ICP-MS) is applied for the quantitative determination of manganese concentration in metal samples. A set of Certified Reference Materials, with different matrix elements (Fe, Cu, Ni, Co, Al) and with certified Mn values, were analyzed. Absorption and plasma break-down depend on the solid phase; as a result, there are differences in mass ablation rates that influence the Mn signal response. In order to be able to compare Mn signals, the relative element-dependent response of tested metal matrices was determined from the ablation of Fe, Cu, Ni, Co, and Al pure metal targets. Relative sensitivity factors (RSFs), calculated as the ratio of the signal intensity of 59Co, 58Ni, 63Cu, and 27Al to the signal intensity of 56Fe, were: 0.79, 0.71, 0.74, and 0.51, respectively. All the isotope signals were matched to the value that should correspond to the relative abundance of 56Fe (91.75%). These experimentally determined RSFs were applied for compensating the observed differences. For quantitative measurements a set of CRM carbon steel samples (SS-451/1 to SS-460/1), with Fe contents of approximately 98%, was used for calibration. The samples with different matrices were measured using the main matrix element as internal standard; for this purpose, the measured intensity was firstly extrapolated, from their content in the sample, to the value that should correspond to a content of 98% and, after, was corrected with the calculated RSF. The developed quantification approach provides manganese results with deviations of 5-10% from the certified values, with relative standard deviations of 3-8%. PMID- 19678977 TI - Pharmacological characterization, localization, and regulation of ionotropic glutamate receptors in skate horizontal cells. AB - Glutamate is believed to be the primary excitatory neurotransmitter in the vertebrate retina, and its fast postsynaptic effects are elicited by activating NMDA-, kainate-, or AMPA-type glutamate receptors. We have characterized the ionotropic glutamate receptors present on retinal horizontal cells of the skate, which possess a unique all-rod retina simplifying synaptic circuitry within the outer plexiform layer (OPL). Isolated external horizontal cells were examined using whole-cell voltage-clamp techniques. Glutamate and its analogues kainate and AMPA, but not NMDA, elicited dose-dependent currents. The AMPA receptor antagonist GYKI 52466 at 100 microm abolished glutamate-elicited currents. Desensitization of glutamate currents was removed upon coapplication of cyclothiazide, known to potentiate AMPA receptor responses, but not by concanavalin A, which potentiates kainate receptor responses. The dose-response curve to glutamate was significantly broader in the presence of the desensitization inhibitor cyclothiazide. Polyclonal antibodies directed against AMPA receptor subunits revealed prominent labeling of isolated external horizontal cells with the GluR2/3 and GluR4 antibodies. 1-Naphthylacetyl spermine, known to block calcium-permeable AMPA receptors, significantly reduced glutamate-gated currents of horizontal cells. Downregulation of glutamate responses was induced by increasing extracellular ion concentrations of Zn2+ and H+. The present study suggests that Ca2+-permeable AMPA receptors likely play an important role in shaping the synaptic responses of skate horizontal cells and that alterations in extracellular concentrations of calcium, zinc, and hydrogen ions have the potential to regulate the strength of postsynaptic signals mediated by AMPA receptors within the OPL. PMID- 19678982 TI - Design of a laser-induced breakdown spectroscopy system for on-line quality analysis of pulverized coal in power plants. AB - It is vitally important for a power plant to determine the chemical composition of coal prior to combustion in order to obtain optimal boiler control. In this work, a fully software-controlled laser-induced breakdown spectroscopy (LIBS) system comprising a LIBS apparatus and sampling equipment has been designed for possible application to power plants for on-line quality analysis of pulverized coal. Special attention was given to the LIBS system, the data processing methods (especially the normalization with Bode Rule/DC Level) and the specific settings (the software-controlled triggering source, high-pressure gas cleaning device, sample-preparation module, sampling module, etc.), which gave the best direct measurement for C, H, Si, Na, Mg, Fe, Al, and Ti with measurement errors less than 10% for pulverized coal. Therefore, the apparatus is accurate enough to be applied to industries for on-line monitoring of pulverized coal. The method of proximate analysis was also introduced and the experimental error of A(ad) (Ash, 'ad' is an abbreviation for 'air dried') was shown in the range of 2.29 to 13.47%. The programmable logic controller (PLC) controlled on-line coal sampling equipment, which is designed based upon aerodynamics, and is capable of performing multipoint sampling and sample-preparation operation. PMID- 19678983 TI - Determination of the hydroxy and carboxylic acid groups in natural complex mixtures of hydroxy fatty acids by 1H nuclear magnetic resonance spectroscopy. AB - The use of trichloroacetyl isocyanate (TAI) to mark both hydroxyl and carboxyl groups borne by the hydrolysis or methanolysis of suberin fragments (a complex mixture of hydroxy fatty acids), allowed the quantitative assessment of the ratio between carboxyl and hydroxy groups, as well as the ratio between primary and secondary hydroxy groups, to be carried out reliably by 1H nuclear magnetic resonance (NMR) spectroscopy. All the samples thus analyzed displayed an excess of CO2H (or CO2CH3) functions with respect to the OH counterparts, albeit to a variable extent, depending on the procedure adopted to isolate the suberin fragments. The precise knowledge of the molar ratio of these two reactive moieties is fundamental for the correct utilization of suberin monomers in polymerization reactions leading to aliphatic polyesters. PMID- 19678984 TI - Analysis of reflectance and transmittance measurements on absorbing and scattering small samples using a modified integrating sphere setup. AB - The aim of the present paper is to analyze reflectance and transmittance measurements on small scattering and absorbing samples. The long term goal is to perform quantitative, spectroscopic in vivo measurements of pigments in small samples of plant material. Small samples such as small leaves constitute a special experimental challenge in cases in which the sample beam has a larger cross-sectional area than the sample. The experimental errors introduced when measuring reflectance and transmittance on small absorbing and scattering samples are investigated theoretically and experimentally by using a blue polyester sample as an appropriate test sample. The experiments are performed with either a mask or a lens setup combined with a mask inserted in the sample beam. In particular, the errors introduced in the reflectance measurements can be very large and larger than 100%. It is shown that any direct illumination of the mask must be avoided. To obtain more accurate values for the reflection coefficient it is necessary to combine the mask with a focusing lens system, adjust the mask and sample very carefully, and choose the ratio between the aperture of the mask and the beam area as large as possible. In the case of transmittance measurements, it is shown that the combination of a special sample fixture and a lens system gives rise to smaller errors compared to the errors introduced by the mask alone or the mask combined with a focusing lens system. PMID- 19678985 TI - Spectroscopic ellipsometry of mucin layers on an amphiphilic diblock copolymer surface. AB - Both visible and infrared (IR) spectroscopic ellipsometry have been employed to study the structure of thin layers of bovine submaxillary mucin (BSM) adsorbed on poly(acrylic acid-block-methyl methacrylate) (PAA-b-PMMA) copolymer and poly(methyl methacrylate) (PMMA) surfaces at three pH values (3, 7, and 10). The adsorbed mucin layer on the copolymer surface had the greatest thickness (17 nm) when adsorbed from a mucin solution at a pH of 3. For the first time, IR ellipsometry was used to identify adhesive interactions and conformational changes in mucin/polymer double layers. After applying the regularized method of deconvolution in the analysis, the formation of hydrogen bonds between the carboxyl groups of the BSM and PAA-b-PMMA copolymer in double layers has been found. The IR ellipsometry data, in agreement with the visible ellipsometry analysis, indicate the pH dependence of adhesion of mucin to the copolymer surface. There is an increase in the amount of hydrogen-bonded carboxyl groups in mucin deposited at a pH of 3. There is no evidence that the amide groups of the mucin participate in this bonding. At the lower pH, the IR ellipsometry spectra after deconvolution reveal an increase in the proportion of beta-sheets in the BSM upon adsorption on the copolymer surface, indicating a more unfolded, aggregated structure. The IR ellipsometry data also indicated some changes in the conformational states of the side groups in the copolymer induced by entanglements and bonding interactions with the mucin macromolecules. Deconvolution provides an unprecedented level of information from the IR ellipsometry spectra and yields important insights. PMID- 19678986 TI - The infrared spectra of Bacillus bacteria part I: vegetative Bacillus versus sporulated cells and the contributions of phospholipids to vegetative infrared spectra. AB - This paper highlights the distinctions between the infrared (IR) absorption spectra of vegetative versus sporulated Bacillus bacteria. It is observed that there are unique signatures clearly associated with either the sporulated or vegetative state and that vegetative cells (or cell debris) can contribute to the spore spectra. A distinct feature at approximately 1739 cm(-1) appears to be unique to vegetative cell spectra and can also be used as an indicator of vegetative cells or cell debris in the spore spectra. The data indicate that the band arises from a lipid-soluble species such as an ester or phospholipid carbonyl bond and are consistent with it being either phosphatidyl glycerol (PG) or phosphatidylethanolamine (PE), two major classes of phospholipids found in vegetative cells of Bacillus species. A companion work discusses bands associated with the sporulated state. PMID- 19678987 TI - The infrared spectra of Bacillus bacteria part II: sporulated Bacillus--the effect of vegetative cells and contributions of calcium dipicolinate trihydrate, CaDP.3H2O. AB - Our previous paper showed that certain infrared (IR) peaks, e.g., the peak at 1739 cm(-1), are due to varying (trace) amounts of vegetative cells amongst the Bacillus spores and that these and other vegetative bands are associated with lipid-soluble compounds, likely an ester or phospholipid. This work investigates the infrared spectra of eight different sporulated Bacillus bacteria. For the endospores it is observed that peaks at 1441, 1277, and 1015 cm(-1) along with a distinct quartet of peaks at 766, 725, 701, and 659 cm(-1) are clearly associated with calcium dipicolinate trihydrate, CaDP.3H2O. It is emphasized that the spore peaks, especially the quartet, arise from the calcium dipicolinate trihydrate and not from dipicolinic acid or other dipicolinate hydrate salts. The CaDP.3H2O infrared peaks and the effects of hydration are studied using quantum chemistry in the PQS software package. The quartet is associated with many modes including contributions from the Ca2+ counterion and hydration waters including Ca-O-H bends, H2O-Ca-O torsions, and O-C-O bends. The 1441 and 1015 cm(-1) modes are planar pyridine modes with the 1441 cm(-1) mode primarily a ring C-N stretch and the 1015 cm(-1) mode primarily a ring C-C stretch. PMID- 19678988 TI - Self-weighted correlation coefficients and their application to measure spectral similarity. AB - A technique for spectral searching with noisy data is described that improves the performance over contemporary approaches. Instead of simply calculating the correlation coefficient between the spectrum of an unknown and a series of reference spectra, greater weight is given to the more intense features in the reference spectra. The weight array, w, is given by |r|/{1 + d}, where the vector r represents the reference spectrum and the difference vector, d, contains the difference between the sample and reference data points, equal to |s-kr|, where k is a scaling factor that eliminates the effect of signal strength. By this approach, a large weight is only given to those points that have relatively high absorbance and are close to their counterparts in the reference spectrum. This technique was shown to give significantly improved performance when applied to noisy spectra of trace atmospheric components obtained by target factor analysis. PMID- 19678989 TI - Quantitative classification of two-dimensional correlation spectra. AB - Two-dimensional (2D) correlation spectroscopy, which takes advantage of the apparent enhancement of spectral resolution, is known to be useful in qualitative discrimination of seemingly similar samples. The possibility of quantitative classification of 2D correlation spectra is even more desirable. Two useful parameters, namely Euclidian distance and correlation coefficient between 2D correlation spectra, are introduced for this purpose. Dry and sweet red wine samples are used to demonstrate the utility of these parameters. The distances between the 2D infrared (IR) spectra of sweet and dry red wines are roughly proportional to the differences of sugar contents in them. The result shows that the two parameters are useful measures for the quantitative evaluation of the similarity among the samples and their corresponding 2D correlation spectra. PMID- 19678990 TI - Study on the crystalline structure transition of syndiotactic polystyrene film during heat treatment by two-dimensional infrared correlation spectroscopy. AB - The crystal structure transition of syndiotactic polystyrene film from the helical conformation to the more stable planar zigzag conformation during a heating process was studied using Fourier transform infrared (FT-IR) spectroscopy in combination with two-dimensional (2D) correlation analysis and perturbation correlation moving-window 2D analysis. The sequence of different conformations during the transition was investigated by analyzing two-dimensional FT-IR correlation spectra in the spectral ranges of 800-700 cm(-1) and 600-500 cm(-1). It was observed that the conformation of delta helical changes prior to gamma helical, and the gamma helical phase is faster than the alpha' planar zigzag phase. By utilizing the 2D asynchronous correlation spectra, the 744 cm(-1) band, which is usually incorporated in the broad 750 cm(-1) band, can now be uniquely attributed as the alpha' zigzag configuration for the first time. Furthermore, by employing thermal perturbation, the shorter helical segments consisting of m = 7 12 and m = 12-20 monomeric units were disturbed in a shorter time than the longer helical segments m = 20-30 during the heating process. PMID- 19678991 TI - Sapphire fiber evanescent wave absorption in turbid media. AB - The influence of particulates on sapphire fiber evanescent wave absorption by water has been studied. Suspensions containing micro-sized graphite flakes and glassy carbon powder were used. Conventional free-space transmittance measurements of these samples showed strong absorption and scattering, which severely screened the absorption by water. However, the absorption on the water band determined from the evanescent wave interaction was unaffected by the presence of the graphite flakes. These results indicate that fiber-optic evanescent wave chemical sensors may be suitable for process control applications involving turbid reactor streams. PMID- 19678992 TI - Fluorescence intensity calibration using the Raman scatter peak of water. AB - Fluorescence data of replicate samples obtained from different fluorescence spectrometers or by the same spectrometer but with different instrument settings can have great intensity differences. In order to compare such data an intensity calibration must be applied. Here we explain a simple calibration method for fluorescence intensity using only the integrated area of a water Raman peak. By applying this method to data from three different instruments, we show that it is possible to remove instrument-dependent intensity factors, and we present results on a unified scale of Raman units. The method presented is a rapid and simple approach suitable for routine measurements with no need for hazardous chemicals. PMID- 19678993 TI - Fourth-order coherent Raman spectroscopy of liquid-solid interfaces: near-surface phonons of TiO2 (110) in liquids. AB - The fourth-order coherent Raman response of a TiO2 (110) surface covered by HCl aqueous solution, neat octanol, acetic acid, or carbon tetrachloride layers is acquired. Four fourth-order optical responses were identified at 837-826, 452 448, 371-362, and 184-183 cm(-1) and assigned to near-surface phonons of TiO2. A third-order response produced in the bulk liquid layer was superimposed on the fourth-order response, when coherent vibrations are efficiently excited in the layer. PMID- 19678994 TI - Fuzzy approach for identifying artistic pigments with Raman spectroscopy. AB - In this work, a fuzzy approach for automatically identifying artistic pigments from their Raman spectra is presented. The uncertainty introduced during the Raman spectrum measurement of pigments is considered in the design of the fuzzy system. The position of the Raman bands in the unknown spectrum can be subject to small displacements due to noise, misalignments in the calibration, etc. Fuzzy logic allows us to work with this uncertainty and to design a system based on the comparison between the Raman band positions in an unknown spectrum recorded from an artwork and the Raman band positions in spectra recorded from reference pigments gathered in databases. The fuzzy system provides the reference pigments whose Raman band positions match those of the unknown pigment analyzed and gives guidance to the decision-making process in the final identification. PMID- 19678995 TI - Fast low frequency (down to 10 cm(-1)) multichannel Raman spectroscopy using an iodine vapor filter. AB - We have constructed a multi-channel Raman spectrometer that is capable of recording the low frequency region down to 5 cm(-1) with a measurement time of a few tenths of a second. An iodine vapor filter, which uses a narrow (approximately 0.03 cm(-1)) absorption line of iodine for Rayleigh scattering elimination, is combined with a multi-channel Raman spectrometer composed of a single polychromator and a charge-coupled device (CCD) camera. Thanks to the high Rayleigh scattering elimination efficiency of the filter, which is over 10(6), Raman spectra of microcrystalline L-cystine from -300 cm(-1) to 1000 cm(-1) are simultaneously measurable with a small gap of 10 cm(-1) (-5 cm(-1) to 5 cm(-1)). Although raw spectra contain many sharp spikes due to the fine structures of iodine absorption, they can be correctly compensated with the use of a transmittance spectrum measured under the same experimental conditions. Many Raman bands including the 9.8 cm(-1) band are measured with a high signal-to noise ratio in both the Stokes and anti-Stokes sides with a measurement time as short as 0.2 s. PMID- 19678997 TI - 6Li atom percentage determination by atomic absorption-emission spectrometry using a natural lithium hollow cathode lamp. AB - A simple method has been developed for the determination of 6Li atom % using combined atomic emission-absorption spectrometry employing a commonly available natural lithium hollow cathode lamp. Unlike in previous practice, there is no need for specially fabricated and high cost 6Li and 7Li monoisotopic lamps in this method. The method requires adjustment of total lithium contents of the sample, i.e., 6Li + 7Li, to 2 microg x mL(-1) based upon atomic emission spectroscopy (AES) (C(aes)) against a 2 microg x mL(-1) natural lithium standard. The concentration of the sample was then analyzed by atomic absorption spectroscopy (AAS) measurements (C(aas)). The difference between the concentration measured by AES and AAS, i.e., C(aes)-C(aas), was calculated. The magnitude of the difference was found to be a function of 6Li fraction in the sample. A calibration curve was constructed by plotting 6Li atom % versus [(C(aes)-C(aas))/C(aes)] x 100. 6Li atom % of an unknown sample can be evaluated by putting its [(C(aes)-C(aas))/C(aes)] x 100 value in the calibration curve. The method is fast, convenient, and precise. PMID- 19678996 TI - Arsenic speciation in biological samples using XAS and mixed oxidation state calibration standards of inorganic arsenic. AB - The speciation of elements without pre-edge features preformed with X-ray absorption near edge structure (XANES) can lead to problems when the energy difference between two species is small. The speciation of arsenic (As) in plant samples was investigated using the mixtures As2S3/As2O5, As2S3/As2O3, or As2O3/As2O5. The data showed that the energy separation (eV) between As2O5 and As2S3 was 5.8, between As2O3 and As2O5 was 3.6, and between As2S3 and As2O3 was 2.1. From the intensity of the white-line feature and the concentration of As species, calibration curves showing a limit of detection of approximately 10% were generated. In addition, an error of +/-10% was determined for the linear combination-XANES (LC-XANES) fitting technique. The difference between the LC XANES fittings and calculations from the calibration curves was <10%. The data also showed that the speciation of As in a sample can be determined using EXAFS (extended X-ray absorption fine structure). Finally, it was also shown that both EXAFS and XANES of the sample should be examined to determine the true speciation of an element. Even though there is a difference of 2 eV between As(III) bound to O and As(III) bound to S, in the EXAFS region the As(III)-S and As(III)-O ligands are clearly visible. However, distinction between the As(III)-O and As(V)-O ligands in the EXAFS spectra was not clearly visible in this study. PMID- 19678998 TI - Near-infrared imaging analysis of cellulose tablets by a band position shift. PMID- 19678999 TI - Bacterial vaccines and the innate immune system: a journey of rediscovery for the allergist-immunologist and all health care providers. PMID- 19679000 TI - Rationale for the clinical use of a ribosome-component immune modulator. AB - Vaccines have long been used to boost the immune system and to confer protection against microbial infections. In contrast, immunotherapy based on ribosomal preparations has been proposed to enhance both specific and nonspecific immune responses (in particular, the mucosal immune defense system) against common respiratory tract pathogens. The aim of this review is to summarize current knowledge on the ability of a combination of four immunogenic ribosomal fractions from different bacteria species and of the extracted fraction from the membrane of Klebsiella pneumoniae to modulate several immune functions. The immunomodulatory activity of the ribosome component is attributed to the presence of highly purified epitopes from cellular fractions obtained by bacterial lysis. The ribosomal preparation is hypothesized to induce a T-dependent immune response consequent network with proliferation of B lymphocytes and production of secretory high-affinity antibodies (in particular, IgA), as well as the creation of a specific immune memory. Indeed, this agent stimulates the activity of macrophages, polymorphonuclear cells, and natural killer cells. The "ideal" vaccine is a highly purified protein bound to a carrier and an adjuvant that enhances the vaccination effect. PMID- 19679001 TI - Ribosome-component immune modulation of respiratory tract infections in adults. AB - The standard therapeutic strategy for bacterial episodes (and bacterial superinfections) is the repeated use of antibiotics. Although antibiotics are fast acting and curative, they do not prevent recurrence or complications. The incidence of recurrent respiratory infections and a substantial increase of costs justify preventive measures based on boosting the patients' immune system by immunomodulation using bacterial products, which, in the long term, stimulates immunity and reduces recurrence of infection. This study was designed to review the epidemiological features of respiratory infections and prevention strategies in adults. A narrative review was performed of the relevant literature. Several products of the immune modulator class are available: bacterial lysates, bacterial membrane component by-products, chemically synthesized compounds, and specific bacterial components (ribosome-component immune modulator [RCIM]). The clinical efficacy of the RCIM has specific and nonspecific immunostimulant properties. Efficacy is mainly documented by decreased recurrent episodes of infection and prescription and need for antibiotics. Benefits are generally observed after 3 months of treatment and are maintained for 6 months. Clinical trial evidence is accruing of the efficacy of RCIM in preventing recurrence of upper and lower airway infections or relapses in adults. PMID- 19679002 TI - Ribosome-component immune modulation of respiratory tract infections in children. AB - More than 25% of infants in their first year of life and 18% of children aged between 1 and 4 years suffer from recurrent respiratory infections (RRIs) in Western countries. Although RRIs are self-limiting as a rule, complications may include otitis media, sinusitis, and bronchial and pulmonary infections. This study was designed to present the available data on immune modulators (defined as drugs that interact with the immune system and modulate immune function by stimulating a more rapid and effective immune response). A ribosome-component immune modulator (RCIM) designed to stimulate both specific and nonspecific immunity in children and thus prevent or alleviate RRI is also described. A narrative review of the literature was performed with a focus on clinical trials. Double-blind, placebo-controlled studies have shown that an RCIM effectively prevents recurrent bronchopulmonary and ear-nose-throat infections; in particular, the number, severity, and duration of infectious episodes and the numbers of antibiotic courses, concomitant medications, and days away from school (children) or the workplace (parents) were reduced. Use of a RCIM is clinically efficacious, incurs minimal risk of adverse events, and, thus, represents a consistent therapeutic approach for RRIs. PMID- 19679003 TI - Safety and tolerability of ribosome-component immune modulator in adults and children. AB - Ribosomal preparations (ribosome-component immune modulators [RCIMs]) do not contain attenuated bacteria and, in contrast to live bacterial extracts, which may induce severe side effects, retain immune stimulating activity without infectious capability. This study was designed to profile the tolerability of RCIM by reviewing narratively all randomized, double-blind, placebo-controlled clinical trials and open-label studies as well as data from postmarketing surveillance studies representing >30 million prescriptions. In the various clinical trials, RCIM tolerability in terms of clinical and laboratory parameters was good. There were no significant differences between patients receiving active treatment or placebo in a survey of tolerability results from randomized, double blind, placebo-controlled studies. Pharmacovigilance analysis does not show a change in the risk profile of RCIM. The only contraindication was correlated with known hypersensitivity to any of the product components. RCIM should not be used in case of acute streptococcal glomerulonephritis, acquired immune deficiency syndrome, severe viral disease, or severe autoimmune disease. Risk associated with the use of RCIM is negligible in recurrent upper and lower airway infections in selected populations, such as children and elderly people. PMID- 19679005 TI - Meta-analysis: making sense of maintenance rituximab? PMID- 19679004 TI - Cost effective analysis of ribosome-component immune modulation. AB - In childhood, recurrent airway infections are the most common cause for medical visits and hospitalizations. Their economic impact is considerable and their treatment accounts for a substantial quota of the health care budget. Despite documented efficacy and safety, ribosomal bacterial vaccine and membrane fraction use is still limited by being prevalently considered for prevention rather than treatment. The objective of this study was to analyze the economic benefit achieved with ribosomal immunotherapy. A review was performed of available literature data on economic parameters. Pharmacoeconomic studies published during the past few years showed how the use of ribosome component immune modulator is capable of reducing expenditure associated with treatment of recurrent respiratory infection and allows achievement of substantial savings in terms of cost, time, and loss of productivity. The initial cost of prevention in patients suffering from chronic respiratory infections is offset by a subsequent saving arising from less recurrence of disease. Direct management costs such as therapy courses, medical visits, medical exams, hospitalizations, etc., and indirect costs such as absence from school and work are not routinely considered by primary care physicians. PMID- 19679006 TI - CCR9:CCL25 in melanoma metastatic to small intestine. PMID- 19679007 TI - Control of experimental melanoma by host CXCR2. PMID- 19679008 TI - Chronic myeloid leukemia in the tyrosine kinase inhibitor era: what is the best therapy? AB - Imatinib mesylate, 400 mg/d, is considered standard therapy for managing patients with chronic myeloid leukemia (CML) in chronic phase, yielding high rates of cytogenetic responses that translate into favorable long-term outcomes. However, some patients do not achieve adequate levels of response, lose a previously acquired response, or are forced to discontinue imatinib therapy because of safety reasons. To avoid these outcomes, several approaches are being tested in the frontline setting, including the use of higher imatinib doses or second generation tyrosine kinase inhibitors such as nilotinib or dasatinib, the latter of which is approved only for managing patients who have imatinib therapy failure. Newer multikinase inhibitors active against multiple ABL1 mutations are also under development for patients in any CML phase who have therapy failure on sequential imatinib and a second-generation tyrosine kinase inhibitor or carry the highly resistant T315I mutation and are not candidates for allogeneic stem cell transplantation. Some of these approaches are expected to improve the outcomes of patients with CML. PMID- 19679009 TI - New agents in acute myeloid leukemia: beyond cytarabine and anthracyclines. AB - The standard therapeutic approaches for acute myeloid leukemia (AML) continue to be based on anthracyclines and cytarabine. However, the prognosis for AML remains poor, especially for patients with high-risk disease. During the past decade, promising novel agents that target DNA replication and repair, as well as cell cycling and apoptosis, have been developed and are being actively investigated in AML. Among these agents is flavopiridol, which interferes with key steps of the cell cycle and effectively promotes cell death, and voreloxin, an intercalating agent that also targets topoisomerase II. Also under clinical study in AML are oligonucleotide antisense constructs, which suppress the translation of proteins essential for leukemic blast survival and proliferation, and agents that target antiapoptotic cascades. In summary, it is hoped that novel therapies such as these will augment and/or supplant our current cytarabine- and anthracycline based approaches, overcome active drug-resistance pathways, and eventually improve outcomes for patients with AML. PMID- 19679010 TI - Incorporating prognostic information into treatment decisions in chronic lymphocytic leukemia. AB - Most patients with chronic lymphocytic leukemia are diagnosed at an early stage, when traditional staging systems fail to distinguish those with an aggressive disease course from those with an excellent prognosis. This failure underscores the need for better prognostic markers. Although many markers have been explored, this review focuses on the newer and most clinically relevant markers: cytogenetic aberrations, zeta-associated protein 70, immunoglobulin mutational status, and CD38 expression. Although these markers have been shown to predict outcomes for groups of patients, individual patients may have a variable course; therefore, the outcome for any one patient remains hard to predict. To date, no study has shown a benefit from early cytotoxic therapy for any subgroup of patients with chronic lymphocytic leukemia. PMID- 19679011 TI - Radioimmunotherapy: strategies for the future in indolent and aggressive lymphoma. AB - The conjugation of radioisotopes to monoclonal antibodies, or radioimmunotherapy (RIT), is a highly active treatment in non-Hodgkin's lymphoma. RIT has demonstrated high response rates and durable remissions in extensively pretreated patients and has proved highly effective as consolidation after induction chemotherapy in the first-line therapy of follicular lymphoma. Early-phase clinical trials have shown highly promising results using RIT as part of conditioning regimens in patients who are to undergo transplantation and as consolidation after chemotherapy in patients with aggressive lymphomas. Recent data suggest that integrating RIT with immunochemotherapy and transplant conditioning regimens may further improve outcomes for patients. PMID- 19679012 TI - Mantle cell lymphoma: are current therapies changing the course of disease? AB - Mantle cell lymphoma (MCL) is a rare entity of non-Hodgkin's lymphoma, although it seems to be increasing in incidence and severity. There is no accepted standard therapy; however, one controlled clinical trial demonstrated that intensive induction immunochemotherapy followed by high-dose radiochemotherapy and autologous stem cell transplantation was superior to conventional treatment. Moreover, uncontrolled studies of intensive immunochemotherapy followed by autologous stem cell transplantation now suggest that MCL may be cured. Insight into the biology of MCL is expanding, opening new avenues of treatment with well defined molecular targets, including CD20, mammalian target of rapamycin, and proteasomes. PMID- 19679013 TI - The future of small molecule inhibitors in lymphoma. AB - For the many patients with lymphoma that has relapsed after and/or has become refractory to existing treatments, the development of novel therapeutics is imperative. Investigation into intracellular processes that are dysregulated during lymphomagenesis has uncovered several new potential targets for anticancer agents. Although monoclonal antibodies and other immunotherapeutics have led to dramatic advances in the treatment of patients with lymphoma, the parallel development of small molecule inhibitors has been equally exciting. These agents, whose small size allows direct entry into tumor cells, can target distinct proteins or complexes, thereby disrupting molecular processes on which neoplastic cells depend for survival and growth. This review surveys the published literature on many of these new targeted molecules, focusing on some of the most promising agents for which phase 2 data currently exist. It also explores the potential for incorporating these agents into broader multidrug regimens. PMID- 19679014 TI - Second-line treatment paradigms for diffuse large B-cell lymphomas. AB - Despite recent major advances in treating diffuse large B-cell lymphoma with dose intense regimens and the addition of the anti-CD20 monoclonal antibody rituximab, a significant proportion of patients will experience early treatment failure, partial response, or relapse after initial chemotherapy. For more than 10 years, the standard treatment for chemosensitive relapses has been based on salvage chemotherapy followed by high-dose therapy and autologous stem cell transplantation in selected patients. However, several important questions remain: What is the best salvage regimen? What is the efficacy of rituximab in an era when R-CHOP (rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone) is accepted as standard care for frontline therapy? What are the risk factors in second-line therapy? What is the best treatment when high-dose therapy and autologous stem cell transplantation are not possible? This article reviews all these issues and discusses new biologic therapies, with the knowledge that improvements in outcome may be achieved through a greater understanding of the biologic parameters associated with poorer prognosis. PMID- 19679015 TI - Betting on immunotherapy for melanoma. AB - Immunotherapy is an effective treatment option for a small percentage of patients with advanced melanoma or at high risk for recurrence after resection of the primary tumor. However, a long period of unsuccessful immune modulation trials involving new cytokines, antibodies, cancer vaccines, adoptive immunotherapy, and combinations generated doubts that benefit could be extended to a larger group of patients. Renewed optimism for the therapeutic potential of immune therapy is currently driven by key advances in tumor immunobiology, including the potential to manipulate and disrupt immune activation checkpoints and tumor defense mechanisms; newer approaches to antigen presentation for immune activation; refinements to procedures for antigen-specific T-cell expansions in vitro and preparative regimens to support their expansion and activity in vivo; gene transfer to alter lymphocyte specificity and function; and the potential for discovery of improved predictive biomarkers to select patients for individual treatments. Proof of concept is provided by durable remissions observed in patients with advanced melanoma enrolled in clinical trials of anti-CTLA-4 and in new studies of adoptively transferred tumor antigen-specific lymphocytes combined with lymphocyte ablation conditioning regimens. Many agents now being developed are predicted to produce broader, more potent, and more effective antitumor immune responses. PMID- 19679017 TI - A survivor's guide for primary care physicians. AB - Building strong relationships among physicians and staff improves the practice's ability to deal with the uncertainties of a rapidly changing environment. Interacting proactively with the economic, social, political, and cultural environment-the practice landscape-provides opportunities for adaptation and ongoing learning. PMID- 19679016 TI - Inside life of melanoma cell signaling, molecular insights, and therapeutic targets. AB - Melanoma is one of the fastest growing tumor types in the United States. Immunotherapy and chemotherapy benefit only a few patients with metastatic disease. Therapy targeting a signaling pathway critical to the cancer's growth can provide dramatic benefit in several other malignancies and may be a valuable strategy for advanced melanoma, if drugs with a favorable therapeutic index are effective against essential molecular pathways. One such target is the V600E "gain-of-function" BRAF mutation found in 60% of melanomas; other mutations or molecular alterations cooperate with V600E BRAF, particularly those that cause loss of function of PTEN, upstream of Akt and mammalian target of rapamycin. Rapid development of new agents, a better understanding of the target pathways and mechanisms of resistance, and carefully designed strategies to optimize combinations and sequences of these agents, potentially with chemotherapy or immunotherapy, may ultimately have the potential to overcome the previously insurmountable obstacle of therapy resistance in melanoma. PMID- 19679018 TI - Use physical therapy to head off this deformity in infants. Consider early PT to prevent severe deformational plagiocephaly. AB - Identify infants with positional preference early and consider referral to pediatric physical therapy at 7 or 8 weeks to prevent severe deformational plagiocephaly. PMID- 19679019 TI - What's growing on your stethoscope? (And what you can do about it). AB - BACKGROUND: Studies have shown that rubbing alcohol pads on stethoscope diaphragms can reduce bacterial colonization, but alcohol pads are used infrequently used and not always available. METHODS: We conducted a prospective, single-blinded study to investigate whether simultaneously scrubbing hands and stethoscope head with alcohol-based hand foam would significantly reduce bacterial counts on the stethoscope. Using their own stethoscope, participants imprinted the stethoscope head onto a chocolate agar plate, then used alcohol based hand foam to cleanse their hands while simultaneously rubbing the stethoscope head. Once the stethoscope heads were dry, the participants imprinted their stethoscope heads onto a second plate. After 48 hours' incubation, we determined the bacterial counts for the prewash and post-wash plates, and compared the 2. RESULTS: We analyzed a total of 184 cultures (from 92 stethoscopes). Both the mean (28 prewash vs 3 post-wash, P=.001) and median (11 prewash vs 1 post-wash, P=.001) colony counts were significantly greater before being cleansed. Three methicillin-resistant Staphylococcus aureus (MRSA) colonies were identified in the prewash period; all were destroyed by the foam. The estimated number of hand washes needed to prevent 1 MRSA colony is 31 (95% confidence interval [CI], 18-89). CONCLUSION: Simultaneously using hand foam to clean hands and stethoscope heads reduces bacterial counts on stethoscopes. Further research is needed to determine whether this intervention can reduce morbidity and mortality associated with bacterial infection. PMID- 19679020 TI - Concussion care: simple strategies, big payoffs. AB - Consider any alteration of mental status that follows a trauma to be a concussion, whether or not there is also a loss of consciousness. Don't order neuroimaging routinely; it is not necessary for diagnosing concussion. Neuroimaging is important, however, for patients who exhibit prolonged unconsciousness, focal neurologic deficits, or worsening symptoms. Treat post concussive headache, a common complaint, with acetaminophen or ibuprofen. PMID- 19679021 TI - Restless legs syndrome: diagnostic time-savers, Tx tips. AB - To diagnose restless legs syndrome (RLS), start with the 4 "essential criteria": (1) a powerful urge to move the legs that is (2) rest-induced, (3) improves with activity, and (4) worsens in the evening. Carefully screen for secondary causes of RLS, including renal failure, pregnancy, iron deficiency, and medications that can cause or exacerbate symptoms. Carbidopa/levodopa is the first-line treatment for patients with intermittent symptoms of RLS; dopamine agonists are recommended for those with daily or refractory symptoms. PMID- 19679022 TI - Glucose control: how low should you go with the critically ill? AB - For hyperglycemic patients admitted to an intensive care unit (ICU), the target blood glucose level should be < or =180 mg/dL, not 81 to 108 mg/dL. More aggressive glucose lowering is associated with a higher mortality rate. PMID- 19679023 TI - Verrucous nodules on the ankle. PMID- 19679024 TI - Clinical inquiries. Are overweight children more likely to be overweight adults? AB - Overweight at any age in childhood increases the risk for overweight in adulthood. The relative risk ranges from 1.9 to 10.1 and increases as children get older. Not all overweight children become overweight adults, however. PMID- 19679025 TI - Clinical inquiries. How useful is a physical exam in diagnosing testicular torsion? AB - A physical exam is useful, but imperfect, in ruling out testicular torsion. The cremasteric reflex or a nontender testicle usually excludes testicular torsion, but case reports have noted the opposite to be true. An abnormal testicular lie can help establish the diagnosis, but occurs in fewer than 50 percent of cases. Other findings are less reliable. The standard of care for diagnosing testicular torsion relies on studies beyond the physical examination. PMID- 19679026 TI - Importance of daily plasma glucose control in type 2 diabetes. PMID- 19679027 TI - When and how to implement basal-bolus therapy: treating to success. PMID- 19679028 TI - Overcoming barriers to insulin replacement. PMID- 19679029 TI - Overview of insulin replacement therapy. PMID- 19679031 TI - EMCDDA publishes updated version of the drug-related infectious diseases testing guidelines in July 2009. PMID- 19679032 TI - Changes in the epidemiology of hepatitis B virus infection following the implementation of immunisation programmes in northeastern Greece. AB - The objective of this study was to investigate changes in the epidemiology of hepatitis B virus infection in the general population and selected groups of immigrants in the region of northeastern Greece over the last decade in relation to the introduction of hepatitis B vaccination programmes. Two population-based seroprevalence surveys were carried out during the years 1992-1994 and 1998-2006. In total, 25,105 individuals were tested for the presence of hepatitis B virus markers: HBsAg, anti-HBs and anti-HBc. Childhood/adolescence immunisation programmes began early in 1994 in selected groups of immigrants and were complemented by the national vaccination programme in 1998. Between 1992-1994 and 1998-2006, the HBsAg carrier rate declined from 5.4% [95% CI: 4.5-5.9] in adults (20-60 years old) and 1.9% [95% CI: 1.6-2.4] in children/adolescents (5-19 years old) of indigenous residents to 3.4% [95% CI: 2.9-3.8] and 0.6% [95% CI: 0.2-1.4] respectively (p<0.05). In spite of a decrease compared with 1992-1994, the percentage of HBsAg carriers was still relatively high in 1998-2006 among the Muslim religious minority group (8.2% [95% CI: 8.0-8.7] in adults and 2% [95% CI: 1.7-2.4] in children/adolescents) and in immigrants from the former Soviet Union (4.3% [95% CI: 3.6-4.7] in adults and 1.1% [95% CI: 0.8-2.4] in children/adolescents) (p<0.05 for both selected groups versus general population). The decline of the prevalence of HBsAg in the general population and selected groups of immigrants in northeastern Greece over the last decade supports the effectiveness of the ongoing immunisation programme although the information on the actual number of cases of acute HBV infection is not available. PMID- 19679033 TI - Vibrio cholerae non-O1 non-O139 infection in an immunocompromised patient returning from Spain, July 2009. AB - We describe a severe gastroenteritis with non-O1, non-O139 Vibrio cholerae in an immunocompromised patient returning from a holiday in Spain in July 2009. Predisposing factors and possible cholera enterotoxin production could explain the unusually grave symptomatology. Patient recovered after doxycyclin treatment. PMID- 19679034 TI - Verocytotoxin-producing Escherichia coli O157 outbreak in Wrexham, North Wales, July 2009. AB - An outbreak of Escherichia coli O157 involving four people in North Wales is currently being investigated. Laboratory typing shows all the isolates belong to phage type 2. All four cases reported eating different products from a fast food outlet in the area. The possibility of other common exposures is being explored. PMID- 19679035 TI - The Swedish new variant of Chlamydia trachomatis (nvCT) remains undetected by many European laboratories as revealed in the recent PCR/NAT ring trial organised by INSTAND e.V., Germany. AB - The May 2009 round of INSTAND's ring trial "Chlamydia trachomatis detection PCR/NAT" included a sample with high amount of the Swedish new variant of C. trachomatis (nvCT). A spectrum of at least 12 different commercial diagnostic nucleic acid amplification tests (NAATs) and many different in house NAATs were applied by the 128 participating laboratories which reported 152 results. Approximately 80% of the results correctly reported the presence of C. trachomatis in the nvCT specimen. The nvCT sample was mainly missed, as expected, by participants using the Roche COBAS Amplicor CT/NG (15.5% of reported results) but also by several participants using in house NAATs. The trend towards using nvCT-detecting NAATs is obvious and in addition to the new dual-target NAATs from Roche and Abbott, and BD ProbeTec ET, also a number of new CE mark-certified commercial tests from smaller diagnostic companies as well as many different in house NAATs were used. Laboratories using commercial or in house NAATs that do not detect the nvCT are encouraged to carefully monitor their C. trachomatis incidence, participate in appropriate external quality assurance and controls schemes, and consider altering their testing system. The reliable detection of low amounts of the wildtype C. trachomatis strain in other samples of the ring trial set indicates a good diagnostic performance of all applied commercial NAATs while also detecting the nvCT strain. PMID- 19679036 TI - What will the next influenza season bring about: seasonal influenza or the new A(H1N1)v? An analysis of German influenza surveillance data. AB - For the next influenza season (winter 2009-10) the relative contributions to virus circulation and influenza-associated morbidity of the seasonal influenza viruses A(H3N2), A(H1N1) and B, and the new influenza A(H1N1)v are still unknown. We estimated the chances of seasonal influenza to circulate during the upcoming season using data of the German influenza sentinel scheme from 1992 to 2009. We calculated type and subtype-specific indices for past exposure and the corresponding morbidity indices for each season. For the upcoming season 2009-10 our model suggests that it is unlikely that influenza A(H3N2) will circulate with more than a low intensity, seasonal A(H1N1) with more than a low to moderate intensity, and influenza B with more than a low to median intensity. The probability of a competitive circulation of seasonal influenza A with the new A(H1N1)v is low, increasing the chance for the latter to dominate the next influenza season in Germany. PMID- 19679037 TI - Epidemiological and transmissibility analysis of influenza A(H1N1)v in a southern hemisphere setting: Peru. AB - We present a preliminary analysis of 1,771 confirmed cases of influenza A(H1N1)v reported in Peru by 17 July including the frequency of the clinical characteristics, the spatial and age distribution of the cases and the estimate of the transmission potential. Age-specific frequency of cases was highest among school age children and young adults, with the lowest frequency of cases among seniors, a pattern that is consistent with reports from other countries. Estimates of the reproduction number lie in the range of 1.2 to 1.7, which is broadly consistent with previous estimates for this pandemic in other regions. Validation of these estimates will be possible as additional data become available. PMID- 19679038 TI - Cases of influenza A(H1N1)v reported in Turkey, May-July 2009. AB - Following the declaration by the World Health Organization (WHO) of human cases of infection with a new influenza A(H1N1)v virus of swine origin, the Turkish Ministry of Health launched a case-based reporting of influenza A(H1N1)v throughout the country on 27 April 2009. The index case was detected on 15 May 2009. As of 17 July 2009 the number of laboratory-confirmed cases of influenza A(H1N1)v totaled 128 of whom 38 were indigenous cases. PMID- 19679039 TI - Cancer Epidemiology: The International Journal of Cancer Epidemiology, Detection and Prevention. PMID- 19679040 TI - Circulating antibodies directed against "polycyclic aromatic hydrocarbon-like" structures in the sera of cancer patients. AB - BACKGROUND: An increase in immunoglobulin (Ig) A isotype directed against benzo(a)pyrene (BP) structure has previously been described in sera of cancer patients. In this study, new polycyclic aromatic hydrocarbon (PAH) conjugates were synthesized in order to more closely mimic the endogenous ligands of the cytosolic aryl hydrocarbon receptor (AhR). PAH [benzo(a)pyrene; 1,2 benzanthracene; dibenz[a,c]anthracene; 7,12-dimethylbenza[a]anthracene; benzo(ghi)perylene] were bound to protein carriers such as bovine serum albumin (BSA) via N-acetyl-cysteine (NAC). METHODS: The levels of circulating antibodies (Abs) directed against PAH-NAC conjugates in the sera of cancer patients were evaluated using an Enzyme-Linked Immunosorbent Assay (ELISA) with these new conjugates. The avidity (IC(50)) and specificity of these circulating Abs were assessed via competition experiments. RESULTS: An increase in Ig directed against these PAH-NAC conjugates was found in the sera of cancer patients, irrespective of the state and stage of the tumors. These Ig were principally of the A isotype. Sera from cancer patients had significantly higher optical density (OD) ranges than the controls, p<0.0001. The ELISA test for breast cancer (n=155) and ovarian cancer (n=62) identified 82% and 92% of positive patients, respectively. The percentage positive in the control group (n=60) was around 5%. Moreover, competition experiments with the different PAH-NAC conjugates and NAC-BSA revealed an estimated avidity of 10(-6)M for the circulating IgA antibodies. CONCLUSIONS: The Abs discriminated between the different PAH-NAC conjugates and NAC-BSA. Therefore, these Abs recognize a carcinogenic PAH-NAC structure and not only a BP structure. These markers may be useful in the future for monitoring cancer evolution and recurrence. PMID- 19679041 TI - Overweight in relation to tumour size and axillary lymph node involvement in postmenopausal breast cancer patients-differences between women invited to vs. not invited to mammography in a randomized screening trial. AB - OBJECTIVES: Overweight is associated with advanced stage at diagnosis in breast cancer patients. This could be explained by specific tumour characteristics or tumour promoting factors in the obese, but a diagnostic delay could also be of importance. Mammographic screening has caused a change towards diagnosis of less advanced tumours. This study investigates invitation to mammographic screening and the association between overweight and tumour size/axillary lymph node involvement at breast cancer diagnosis in postmenopausal women. METHODS: In 1976 a randomized mammographic screening trial, inviting 50% of all women aged 45-69 was set up in Malmo, Sweden. The present analysis examined overweight (body mass index >or=25) as a determinant for large tumours (>20mm) and axillary lymph node involvement in postmenopausal women. These associations were studied separately in patients diagnosed prior to the mammographic screening trial, in invited women and in non-invited subjects (controls). In all, 2478 postmenopausal women were diagnosed with invasive breast cancer in these groups between 1961 and 1991. Logistic regression analysis allowed adjustment for other potential determinants of tumours size and axillary lymph node involvement. RESULTS: In women diagnosed before the onset of the screening trial and in women not invited to mammography in the trial (controls), overweight was positively associated with large tumour size and axillary node involvement. There was no statistically significant association between overweight and these factors in women invited to mammographic screening. CONCLUSION: Invitation to mammographic screening may be particularly important for overweight postmenopausal women in order to detect breast tumours early. PMID- 19679042 TI - Remnant lipoproteins induced proliferation of human prostate cancer cell, PC-3 but not LNCaP, via low density lipoprotein receptor. AB - BACKGROUND: Hypertriglyceridemia has been shown to be one of the risk factors for prostate cancer. In this study, we investigated the effect of remnant lipoproteins on cell growth in prostate cancer cell lines. METHODS: Remnant lipoproteins were isolated as remnant like particles (RLP) from human plasma. We used RLP for TG-rich lipoproteins and low density lipoproteins (LDL) for cholesterol-rich lipoproteins respectively and examined the effect of lipoproteins on proliferation of PC-3 and LNCaP cells using MTS assays. Moreover, we studied the effect of RLP and LDL treatment on the regulation of lipoprotein receptors in prostate cancer cells to investigate the relationship between lipoprotein-induced cell proliferation and lipoprotein receptor expression using real-time PCR, Western blotting assays and siRNA. RESULTS: RLP effectively induced PC-3 cell proliferation more than LDL, whereas both RLP and LDL could not induce LNCaP cell proliferation except at a higher concentration of RLP. LDL receptor (LDLr) was expressed in both prostate cancer cells but there was a sharp difference of sterol regulation between two cells. In PC-3 cells, LDL decreased the LDLr expression in some degree, but RLP did not. Meanwhile LDLr expression in LNCaP was easily downregulated by RLP and LDL. Blocking LDLr function significantly inhibited both RLP- and LDL-induced PC-3 cell proliferation. CONCLUSIONS: This study demonstrated that RLP-induced PC-3 cell proliferation more than LDL; however, both RLP and LDL hardly induced LNCaP cell proliferation. The differences of proliferation by lipoproteins might be involved in the regulation of LDLr expression. PMID- 19679043 TI - African American-preponderant single nucleotide polymorphisms (SNPs) and risk of breast cancer. AB - BACKGROUND: African American women more often present with more aggressive types of breast cancer than Caucasian women, but little is known whether genetic polymorphisms specific to or disproportionate in African Americans are associated with their risk of breast cancer. METHODS: A population-based case-control study was conducted including 194 cases identified through the Metropolitan Detroit Cancer Surveillance System and 189 controls recruited through random digit dialing to examine polymorphisms in genes involved in estrogen metabolism and action. RESULTS: The African American-specific CYP1A1 5639C allele was associated with an increased risk of breast cancer (odds ratio (OR)=2.34, 95% confidence interval (CI) 1.23-4.44) and this association with the CYP1A1 5639 locus was dependent on another polymorphism in the CYP3A4 gene (P=0.043 for the interaction). In addition, African American-predominant CYP1B1 432 Val allele was significantly more often found in the cases than in the controls overall and the HSD17B1 312 Gly allele was specifically associated with premenopausal breast cancer risk (OR=3.00, 95%CI 1.29-6.99). CONCLUSION: These observations need to be confirmed in larger studies due to the limited statistical power of the study based on a small number of cases. PMID- 19679044 TI - A mini-review of familial ovarian germ cell tumors: an additional manifestation of the familial testicular germ cell tumor syndrome. AB - INTRODUCTION: While testicular germ cell tumors (TGCTs) are the most common malignancy in young men, germ cell tumors in women are uncommon. Familial clustering, epidemiologic evidence of increased risk with family or personal history of TGCT, and associations with genitourinary tract anomalies suggest an underlying genetic predisposition to TGCT, but traditional linkage studies have yet to identify a highly penetrant TGCT cancer susceptibility gene. In this paper, we investigate the familial occurrence of testicular and ovarian germ cell tumors. METHODS: We report a family in which a TGCT and an ovarian germ cell tumor (OGCT) occurred in two siblings, summarize the existing literature on familial occurrences of OGCT, either alone or in combination with extragonadal or TGCTs, and compare the incidence of familial and sporadic testicular and ovarian GCTs. Sporadic GCT data were obtained from the US Surveillance Epidemiology and End Results (SEER) registry. RESULTS: We identified 16 reports of OGCT occurring in conjunction with either ovarian, testicular or extragonadal GCT. In these familial cases, the mean age at onset of female dysgerminoma was younger than that noted in the general population (age 17 vs. age 24, p=0.01). In SEER, the incidence of TGCT was 15 times higher than that of OGCT. Histologic distributions in males and females showed distinctly different patterns. DISCUSSION: Although the incidence of OGCTs in the general population is quite low, its occurrence in multiple members of the same family and in families with TGCT suggests that a gene conferring susceptibility to GCTs may exist in some families. PMID- 19679045 TI - A polymorphic variant of the insulin-like growth factor type I receptor gene modifies risk of obesity for esophageal adenocarcinoma. AB - BACKGROUND: To investigate potential biologic mechanisms underlying the association between obesity and risk for esophageal adenocarcinoma (EADC), we studied the frequency of a common polymorphism of the insulin-like growth factor I receptor (IGF-IR) gene in patients with either gastroesophageal reflux disease (GERD), premalignant Barrett esophagus (BE) and or invasive EADC. METHODS: Using a well characterized series of 431 individuals enrolled in a case-control study, we studied the frequency of the IGF-IR gene polymorphism, G1013A. RESULTS: On multivariate analysis controlling for age and gender, in comparison to asymptomatic controls, obese individuals with the polymorphic A-variant (G/A, A/A) were found to have significantly increased risk for EADC (OR 4.81; 95%CI 1.09-21.15), whereas obese individuals with the G/G variant were not at statistically significant increased risk (OR 2.69; 95%CI 0.41-17.62). Similarly, compared to asymptomatic controls, only obese individuals with the A-variant (G/A, A/A) were at increased risk for BE (OR 3.11; 95%CI 1.12-8.63), while obese individuals with the G/G variant were not at increased risk for BE (OR 2.91; 95%CI 0.69-12.15). CONCLUSION: We conclude that the common IGF-IR gene polymorphism G1013A modulates the risk of obesity for EADC, an effect most likely mediated by altered the receptor function by influencing gene transcription or mRNA stability. These findings further implicate the insulin-like growth factor axis in the molecular pathogenesis of EADC, and represent a plausible mechanistic link underlying the association between obesity and malignancy. PMID- 19679046 TI - Antioxidant activity in essential oils of Cnidium officinale makino and Ligusticum chuanxiong Hort and their inhibitory effects on DNA damage and apoptosis induced by ultraviolet B in mammalian cell. AB - BACKGROUND: Owing to their high volatile aroma, the dried rhizomes of Cnidium officinale (C. officinale) and Ligusticum chuanxiong (L. chuanxiong) are used as herbal drugs to treat blood pressure depressant, a deficiency disease of antivitamin, inhibition of small intestine sympathetic nerve and as cosmetics for skin care. However, little has been known about the protective effect of their essential oils against ultraviolet B (UVB)-induced DNA damage. METHODS: In this study, we report antioxidant activity of their essential oils using DPPH and ABTS scavenging assay. In addition, the composition of essential oils was measured by GC/MS. We also investigated whether these essential oils could inhibit UVB induced DNA damage and apoptosis in the mammalian cell using intracellular DNA migration and expression level of phospho-H2A.X. RESULTS: Twenty constituents in the essential oil were identified and they showed good antioxidant properties, in that IC(50) value in DPPH and ABTS showed 6.79 and 7.33microg/ml and 1.58 and 1.58microg/ml in C. officinale and L. chuanxiong. Their treatment inhibited the migration of damaged DNA induced by UV-B; furthermore, they decreased p21 expression and increased cyclin D1 expression as apoptosis-regulatory genes. CONCLUSIONS: These results suggest that essential oils in C. officinale and L. chuanxiong may exert inhibitory effects on DNA damage and apoptosis induced by UVB through their high free radical scavenging ability. PMID- 19679047 TI - Detection of p16 promotor hypermethylation in "Maras powder" and tobacco users. AB - BACKGROUND: A plant powder called "Maras powder" is widely used instead of cigarette smoking in the South-Eastern region of Turkey. It has been confirmed that this powder comprises tobacco Nicotiana rustica L. METHODS: The aim of this study was to assess the effect of Maras powder and cigarette smoking on the P16 promotor hypermethylation. Twenty-two Maras powder users (Group I), 12 cigarette smokers (Group II), and 16 healthy controls who neither smoked nor used Maras powder (Group III) were included in the study. Hypermethylation of the P16 gene was examined using methylation-specific PCR (MSP) method in the blood of the three groups. RESULTS: Aberrant P16 methylation was found in 7 of the 22 (31.8%) in Group I, in 3 of 12 (25%) in Group II, and in 1 of 16 (6.25%) in Group III. CONCLUSION: Maras powder may be as harmful as cigarette smoking, leading to hypermethylation in P16 and warrants detailed studies on this subject. PMID- 19679048 TI - Prepubertal octylphenol exposure up-regulate BRCA1 expression, down-regulate ERalpha expression and reduce rat mammary tumorigenesis. AB - BACKGROUND: Endogenous estrogens play an important role in the development of breast cancer. Octylphenol (OP) and genistein (GEN) are estrogen-like chemicals. Prepubertal estradiol and genistein exposure can up-regulate BRCA1 mRNA in mammary gland and reduce futuer breast cancer risk. In the present study, the effects of prepubertal exposure to high-dose OP and GEN on mammary carcinogenesis and the association with the expression of BRCA1 and ERalpha were investigated. METHODS: Prepubertal female Sprague-Dawley rats were exposed to 20, 40, 80mg/kg OP daily from postnatal day (PND) 22-28, subsequently, the rats were given a single dose of 100mg/kg 7,12-dimethylbenz [a] anthracene (DMBA) on PND42 to induce mammary tumor. RESULTS: The incidence of DMBA-induced mammary tumors significantly decreased when rats were treated with 40mg/kg OP. BRCA1 mRNA and protein expression were found up-regulated and ERalpha expression was down regulated in the mammary tumor when rats were exposed to 40mg/kg octylphenol. CONCLUSION: Exposure 40mg/kg octylphenol can reduce later breast cancer risk in prepubertal Sprague-Dawley rats, the protective effect of OP is associated with persistent up-regulation of BRCA1 and down-regulation of ERalpha in the mammary tumor. PMID- 19679049 TI - Hospital-based prevalence and sensitivity of high-risk human papillomavirus in Thai urban population. AB - BACKGROUND: To study the prevalence of high-risk human papillomavirus (HPV) types 16, 18, 31 and 33 in healthy Thai women using polymerase chain reaction (PCR) technique. MATERIAL AND METHOD: Two hundred and sixty three healthy urban women in Pathumthani, Thailand were recruited. Cervical cancer screening was performed and residual specimen from Pap smears was subjected to PCR to identify the presence of HPV types 16, 18, 31 and 33. Individuals' demographic, health specific and sexual behavior data were also collected. RESULTS: Colposcopic biopsy revealed cases with high-grade squamous intraepithelial lesions (HSIL) and low-grade squamous intraepithelial lesions (LSIL). HPV 16 and 18 positive results were associated with abnormal Pap smears. Genotyping gave a 6.1%, 11.8%, 12.1%, and 14.1%, prevalence for HPV types 16, 18, 31 and 33, while 25% were infected with multiple HPV types. DISCUSSION: High-risk HPV screening, used with abnormal pathology of HSIL gives 100% sensitivity and negative predictive value. Data from patient showed not significant correlation with neither different religion level of education, marital status, age of first sexual experience nor the number of sexual partners. Thus high-risk HPV screening is a recommended procedure with excellent sensitivity for detecting HSIL. PMID- 19679050 TI - Estrogenic activity of bovine milk high or low in equol using immature mouse uterotrophic responses and an estrogen receptor transactivation assay. AB - BACKGROUND: Cow's milk contain phytoestrogens especially equol depending on the composition of the feed ration. However, it is unknown whether milk differing in equol exhibits different estrogenicity in model systems and thereby potentially in humans as milk consumers. METHODS: The estrogenicity of high and low equol milk (HEM and LEM, respectively) and purified equol was investigated in immature female mice including mRNA expression of six estrogen-sensitive genes in uterine tissue. Extracts of HEM and LEM were also tested for estrogenicity in vitro in an estrogen receptor (ER) reporter gene assay with MVLN cells. RESULTS: The total content of phytoestrogens was approximately 10 times higher in HEM compared with LEM, but levels of endogenous milk estrone and 17beta-estradiol were similar in the two milk types (503-566 and 60-64.6pg/ml, respectively). There was no difference in uterine weight between mice receiving LEM and HEM, and no difference from controls. Equol (50 times the concentration in HEM) was not uterotrophic. The ERbeta mRNA expression was down-regulated in the uteri of HEM mice compared with LEM and controls, but there was no difference between milk types for any of the other genes. Extracts of HEM showed a higher estrogenicity than extracts of LEM in MVLN cells, and there was a dose-dependent increase in estrogenicity by equol. CONCLUSION: The higher in vitro estrogenicity of HEM was not reflected as a higher uterine weight in vivo although the down-regulation of ERbeta in uterine tissue of HEM mice could suggest some estrogenic activity of HEM at the gene expression level. PMID- 19679051 TI - Fibrous pseudotumors of tunica albuginea, tunica vaginalis and epididymis: report of two cases. AB - We describe two rare cases of fibrous pseudotumor of the paratesticular region. In the first case, five nodules arising from the tunica albuginea of right testicle causing scrotal, enlargement raising after urinary tract infection were seen. In the second case, multiple nodules arising tunica albuginea, tunica vaginalis and epididymis raising after left varicocelectomy operation were observed. The histology showed a paucicellular fibroblastic proliferation of cells within a hyalinized collagenous fibrous stroma containing numerous thin walled blood vessels accompanied by lymphocytes and plasma cells in tumor tissues in both cases. Tumors in both cases were successfully resected. After operation, both patients had an uneventful recovery without any complications. PMID- 19679052 TI - Immunohistochemical analysis of selected molecular markers in esophagus precancerous, adenocarcinoma and squamous cell carcinoma in Iranian subjects. AB - BACKGROUND: The molecular and cellular mechanisms linking chronic inflammation and gastrointestinal malignancy are not known with certainty. AIM: To investigate changes in potential causative factors during progression of esophagus cancer in a population living in high-risk area in Iran. SUBJECTS: Formalin-fixed, paraffin embedded esophageal specimens (n=87) from patients with gastroesophageal reflux disease (GERD), Barrett's metaplasia, adenocarcinoma (ADC) and squamous cells carcinoma (SCC) were collected based on their pathological diagnosis. METHODS: Immunohistochemical (IHC) technique was used to study tissue accumulation of P53, P21, cyclooxygenase-2 (COX-2), glutathione S-transferase-P (GST-Pi) and nitrotyrosine (NT) in patients and controls. RESULTS: P53 expression was not detected in esophageal tissues from normal and GERD samples, whereas it was found positive in Barrett's, ADC, and SCC samples. P21 positive sample was relatively higher in ADC patients as compared to that in SCC (ADC: 52.6%; SCC: 25%). GST-Pi expression was equally accumulated in all the samples. NT was predominantly expressed in ADC (72.7%). COX-2 expression was significantly higher in Barrett's (60.0%) and ADC (66.6%) as compared to that in GERD, SCC and normal. These data were further confirmed by detecting the scores of immunostainings in all the positive samples. CONCLUSION: The pathological changes in ADC and SCC samples which were associated with increasing frequency of NT and COX-2 provides further evidence for involvement of these inflammatory factors in progression of esophagus cancer. PMID- 19679053 TI - Education and cancer incidence in a rural population in south India. AB - BACKGROUND: Population-based studies describing the association between education and cancer incidence has not yet been reported from India. METHODS: Information on the educational attainment of 4417 cancer cases aged 14 years and above, diagnosed during 2003-2006 in Dindigul district, Tamil Nadu, India, was obtained from the Dindigul Ambilikkai Cancer Registry, which registers invasive cancer cases by active methods from 102 data sources. Population distribution by 5-year age groups and for four educational levels namely no education, education 12 years, was obtained from census data. Standardized rate ratios based on age-standardized rates were calculated to study cancer risks for different educational levels. RESULTS: Men and women with no education had higher overall cancer incidence rates compared to the educated population. The risk of cervix, mouth, esophagus, stomach and lung cancers were inversely associated with higher levels of education whereas a high incidence of breast cancer was observed with increasing educational levels. The standardized rate ratio of cervical cancer 0.32 (95% CI: 0.19-0.52) and of breast cancer was 6.08 (95% CI: 1.81 20.48) for women with more than 12 years of education compared to those with no education. There was paucity of cases in the highest education level for most cancers. CONCLUSION: With more and more women in rural India becoming educated, one could foresee breast cancer becoming more frequent even in rural areas of India in future. PMID- 19679054 TI - Cancer mortality and occupational exposure to aromatic amines and inhalable aerosols in rubber tire manufacturing in Poland. AB - AIM: Most data on carcinogenic risk in the rubber industry are based on data from Western countries. This study assessed cancer risks in a retrospective cohort in a Polish tire manufacturing plant, relying on quantified exposure to inhalable aerosols and aromatic amines instead of job titles or external comparisons. METHODS: Cumulative exposure for all exposures was assigned to cohort members based on estimates from a company-specific JEM. Cancer risks associated with cumulative exposure adjusted for co-exposures, gender and year of birth were calculated. RESULTS: Exposure levels were higher for women than for men. Aromatic amine exposure was significantly associated with increased urinary bladder cancer risk (RR=7.32-8.27), depending on exposure level, and prostate cancer at low levels only (RR=5.86). In women, increased risks were found for all cancers (RR=2.50) and of the digestive organs and peritoneum (RR=4.54) at low level only, while an exposure-response association with breast cancer risk was found. Inhalable aerosol exposure was associated with cancers of the liver and intrahepatic bile ducts in a dose-dependent manner, while dose-dependent reduced risks were found for respiratory cancers (most notably the larynx) and cancer of the colon. CONCLUSIONS: Increased risks for specific cancer sites in this rubber plant were similar to Western Europe and the US. However, several cancer risks were gender-specific which could relate to higher exposure levels in women or to differences in exposures to chemicals not assessed in this study. PMID- 19679055 TI - Vitamin D receptor variants and the malignant melanoma risk: a population-based study. AB - BACKGROUND: There is continuing interest in identifying low-penetrance genes which are associated with an increased susceptibility to common types of cancer, including malignant melanoma. METHODS: We sought to examine the association between four VDR common variants (rs1544410, rs731236, rs10735810, rs4516035) and the risk of melanoma in the Polish population. We also determined the prevalence of compound carriers of VDR and known MM genetic risk factors MC1R and CDKN2A (A148T) variants. We examined 763 unselected melanoma cases, 763 healthy adults matched for sex and age with the melanoma cases and 777 newborns. RESULTS: None of the VDR variants alone or as compound carriers of two or more of the VDR genotypes were associated with MM risk. There were no major differences between the prevalences of the examined variants among patients with MM on UV-exposed and UV-non exposed skin areas, as well as among early-onset and late-onset cases. We found no association between VDR and MC1R or between VDR and CDKN2A common variants. A statistically significant over-representation of one VDR haplotype: rs731236_A+rs1544410_T (OR=3.2, p=0.02) was detected. Linkage disequilibrium of rs1544410 and rs731236 was confirmed. CONCLUSION: To answer the question, whether VDR can be regarded as melanoma susceptibility gene, additional, large multi center association studies have to be performed. PMID- 19679056 TI - Natural history of esophageal and gastric cardia precursor by repetitive endoscope screening with 425 adults in a high-risk area in China. AB - OBJECTIVE: To observe the natural history and to determine appropriate screening interval for esophageal and gastric cardia precursors. METHODS: Repetitive endoscopic screenings were performed among 425 forty to sixty-nine-year-old subjects in a high-risk region in Northern China. RESULTS: We observed 8 subjects develop severe dysplasia (SD), another 8 develop carcinomas in situ (Cis), and 4 develop invasive cancer. The time and baseline diagnosis (BD) for the 8 SD subjects were: 13 months after normal epithelium in one case, 7 months after base cell hyperplasia (BCH) in another case, 3, 4, 4, and 10.5 months after mild dysplasia (mD) in four cases, and 12.5 and 43.4 months after moderate dysplasia (MD) in two cases. The time and BD for the 8 Cis cases were: 18 and 51.7 months after BCH in two cases, 48 months after mD in one case, 4 and 13 months after MD in two cases, and 3.5, 9, and 17.5 months after SD in the other three cases. The time and BD for the four invasive cancer cases were, 13 months after mD in one case, 50 months after MD in another case, and 14 and 19 months after SD in two cases. In addition to natural history observation, we also found sex, age (over 50 vs under 50), family history of upper gastrointestinal cancer, and the presence of multiple esophageal lugol-void lesions to be significant predicators for dysplastic progression; the corresponding OR (95% CI) and P-value were 1.98 (1.14-3.46) and 0.02; 2.32 (1.29-4.19) and 0.004; 1.81 (1.06-3.11) and 0.03; and 4.67 (2.70-8.06) and 0.000 respectively. CONCLUSIONS: A 5-year screening interval for BCH and mD, and a 3-year interval for MD, as suggested in China in 2005, may be too long for rapid developing precursors or individuals at high-risk for dysplastic progression. Therefore, shorter intervals should be considered. PMID- 19679057 TI - Patterns of HER2 testing in the management of primary breast cancer. AB - BACKGROUND: Women with invasive breast cancer should be tested for human epidermal growth factor receptor-2 (HER2) status at the time of diagnosis. To date, no population-based patterns of use studies have examined demographic and clinicopathologic factors associated with decisions by clinicians to test patients. METHODS: We reviewed summary pathology reports submitted to the Connecticut Tumor Registry for all Black/African American (B/AA) women (n=644) and a 7% random sample (n=720) of White women diagnosed in 2000-2003 with primary invasive breast carcinoma. Receipt of a HER2 test (yes vs. no) was examined in relation to patient race, age, socioeconomic status, year of diagnosis, estrogen receptor (ER) status, tumor grade, lymph node status, size and stage at diagnosis. RESULTS: A greater proportion of tumors from B/AA patients were tested compared to those of White women (69.5% vs. 61.9%, p<0.05). Tumors of patients under the age of 60 were 1.50-times more likely than older women to have been tested, and B/AA women were 1.40-times more likely than White patients to be tested. HER2 testing was more likely to be observed when information also was reported about ER status (OR=15.9, p<0.001), tumor grade (OR=2.28, p<0.05), tumor size (OR=2.16, p<0.05), and lymph node status (OR=2.06, p<0.05). CONCLUSIONS: Variation in which breast cancer patients received HER2 testing appears to reflect expectations about a woman's prognosis. Discrepancies in receipt of testing deserve further study as current guidelines call for all tumors to be assessed in order to adequately characterize prognosis and determine eligibility for HER2-targeted therapy. PMID- 19679058 TI - Human papillomavirus in oral leukoplakia is no prognostic indicator of malignant transformation. AB - BACKGROUND: Oral leukoplakia is considered as a premalignant lesion for the development of oral squamous cell carcinoma (OSCC); several risks factors have been reported to contribute to this step-wise carcinogenesis; including human papillomavirus (HPV). Nevertheless, few reports have analyzed both the HPV status and the genotype in a single individual who develops OSCC from pre-existing oral leukoplakia. In this study, we surveyed the HPV status, genotype and clinicopathological risk factors in cases of malignant transformation from pre existing oral leukoplakia. METHODS: HPV genomic DNA was detected by PCR (MY09/MY11 in conjugation with nested primer-GP05+/GP06+) from paraffin sections, and the genotype was determined by direct DNA sequencing. Fisher's exact test and logistic regression were used to analyze risk factors for malignant transformation of oral cavity leukoplakia. RESULTS: One hundred and sixty-seven patients with oral leukoplakia were enrolled; including 12 who had malignant transformation from the pre-existing oral leukoplakia. HPV prevalence was 22.8% in cases with oral leukoplakia. The risk factor associated with malignant transformation was recurrence of leukoplakia after treatment (p=0.03), nevertheless, HPV status was not statistically significant by logistic regression analysis. Among these 12 patients with malignant transformation from pre-existing oral leukoplakia, the status or genotype of HPV was chaotic; the oral habits of these patients might contribute to malignant transformation. CONCLUSIONS: Our data suggest that HPV in oral leukoplakia is no prognostic indicator of malignant transformation. PMID- 19679060 TI - Overexpression of the Interferon regulatory factor 4-binding protein in human colorectal cancer and its clinical significance. AB - BACKGROUND: IFN regulatory factor 4-binding protein (IBP) is a novel type of activator of Rho GTPases. It has been linked with differentiation and apoptosis of lymphocytes, but its function in oncogenesis remains unclear. Here we studied the expression of endogenous IBP in four human colorectal cancer cell lines, normal, adenoma and tumor colorectal tissues. METHODS: Molecular (Western blot and RT-PCR), and confocal analyses were used to investigate IBP expression in human colorectal cancer cell lines. Matched normal and tumor tissue sections of 63 patients and 15 adenoma tissue sections were analyzed for IBP expression by immunohistochemistry (IHC). RESULTS: IBP was ubiquitely expressed in human colorectal cancer cell lines. The expression of IBP can be detected at both the mRNA and protein level in SW480, SW620 and HT29 cells. Clinically, IBP were elevated in human colorectal cancer specimens in comparison to normal colorectal tissues. Substantial high expression of IBP was observed in colorectal cancer tissues (67%), whereas corresponding normal tissues and 15 adenoma tissues showed consistently absent immunoreactivity of IBP. Moreover, IBP expression is correlated with the differentiation level of colorectal cancer cells (p<0.05) and clinical stage of patients (p<0.01). CONCLUSIONS: Our data show, for the first time, a dysregulated expression of IBP in human colorectal cancer, offering new perspectives for its role in cancer development and progression. IBP may be a novel tumor marker and a therapeutic target for colorectal cancer. PMID- 19679059 TI - A gene marker panel covering the Wnt and the Ras-Raf-MEK-MAPK signalling pathways allows to detect gene mutations in 80% of early (UICC I) colon cancer stages in humans. AB - BACKGROUND: Very recently a gene marker panel that allows the mutational analysis of APC, CTNNB1, B-RAF and K-RAS was conceived. The aim of the present study was to use the 4-gene marker panel covering the Wnt and Ras-Raf-MEK-MAPK signalling pathways to determine the percentage of sporadic colorectal carcinomas (CRC) carrying at least one of the four above-mentioned genes in a mutated form alone and/or in combination with microsatellite instability (MSI) and to compare the sensitivity of the gene marker panel used in this study with that of gene marker panels previously reported in the scientific literature. METHODS: CTNNB1 and B RAF were screened by PCR-single-strand conformation polymorphism analysis and K RAS gene mutations by restriction fragment length polymorphism analysis. For the mutational analysis of the APC gene mutation cluster region (codons 1243-1567) direct DNA sequencing was performed. The U.S. National Cancer Institute microsatellite panel (BAT25, BAT26, D2S123, D5S346 and D17S250) was used for MSI analysis. RESULTS: It could be shown that about 80% of early stage CRC (UICC stages I and II) and over 90% of CRC in the UICC stage IV carried at least one mutated gene and/or showed MSI. No significant increase in the gene mutation frequencies could be determined when comparing tumours in the UICC stage I with those in UICC stage IV. CONCLUSIONS: When compared with previously published gene marker panels the 4-gene marker panel used in the present study shows an excellent performance, allowing to detect genetic alterations in 80-90% of human sporadic CRC samples analyzed. PMID- 19679061 TI - Survivin and its spliced isoform gene expression is associated with proliferation of renal cancer cells and clinical stage of renal cancer. AB - BACKGROUND: Survivin has been implicated in inhibition of apoptosis. To date, alternatively spliced isoforms, Survivin-2alpha, -2B, -DeltaEx3, -3B, have been described. We assessed the effect of survivin gene expression on the proliferation of renal cancer (RCC) cells, and studied the association of survivin and its spliced isoform gene expression levels with the clinical stage of RCC. METHODS: Gene expression of survivin and its spliced isoform in RCC cells, Caki-1, were performed by RT-PCR. We knocked down the gene expression of Survivin using small interfering RNA (siRNA), and assessed the cell proliferation by MTS assay. Next, we quantified the gene expression levels of survivin and its isoform in nephrectomy samples using quantitative real-time PCR. RESULTS: In Caki 1 cells, survivin and survivin-2alpha, -2B were expressed higher than survivin DeltaEx3. Decrease of Survivin gene expression by transfection of siRNA was accompanied by inhibition of the proliferation of Caki-1 cell with 36% decrease in comparison with negative control transfected cells (p<0.01). In clinical RCC tissues, survivin expression levels in metastatic stage were significantly higher compared with those in distant metastasis stage (M0:M1=1:4.81, p=0.014); survivin 2B gene expression levels in pT3 tumors were associated significantly higher than those in pT1 (pT1:pT3=1:4.50, p=0.043). No significant differences were found in survivin-2alpha expression levels and the ratio of survivin-2B/survivin gene expression levels among any clinical stages. CONCLUSION: We first demonstrated the gene expression of survivin-2alpha in renal cancer cells, and also showed that survivin and its spliced isoforms had associations with renal cancer cell proliferation and distant metastases. PMID- 19679062 TI - MDM2, P53, P21WAF1 and pAKT protein levels in genesis and behaviour of adenoid cystic carcinoma. AB - BACKGROUND: MDM2, P53, P21(WAF1) and pAKT are proteins associated with the balance between cell death and survival. There are many hypotheses regarding the role of these proteins in salivary gland tumours. However, many molecular events that activate or inactivate regulatory genes remain unknown. The aim of this study was to evaluate and to correlate MDM2, P53, P21(WAF1) and pAKT protein expressions in adenoid cystic carcinomas (ACC). METHODS: Twenty-two cases of ACC were evaluated by immunohistochemistry and one cell line derived from ACC was analyzed by Western Blotting and immunofluorescence techniques. RESULTS: Strong MDM2 and pAKT, variable P53 and null P21 expressions were found in the cases analyzed, but no statistical correlation was established when comparing MDM2 and pAKT expressions in the 3 different ACC subtypes. The ACC cell line showed intense nuclear and cytoplasmatic MDM2 and pAKT expressions and null P53 and P21 expressions. CONCLUSIONS: Results indicate that MDM2 and pAKT are related to the tumorigenesis of ACC, but they might not be directly connected to tumour progression. We also demonstrate that the pAKT pathway is active in ACC and it seems to be activating the MDM2 shuttle from the cytoplasm to the nucleus, where it phosphorylates P53 and carries it to the cytoplasm for degradation. PMID- 19679063 TI - Novel generating protective single nucleotide polymorphism barcode for breast cancer using particle swarm optimization. AB - BACKGROUND: High-throughput single nucleotide polymorphism (SNP) genotyping generates a huge amount of SNP data in genome-wide association studies. Simultaneous analyses for multiple SNP interactions associated with many diseases and cancers are essential; however, these analyses are still computationally challenging. METHODS: In this study, we propose an odds ratio-based binary particle swarm optimization (OR-BPSO) method to evaluate the risk of breast cancer. RESULTS: BPSO provides the combinational SNPs with their corresponding genotype, called SNP barcodes, with the maximal difference of occurrence between the control and breast cancer groups. A specific SNP barcode with an optimized fitness value was identified among seven SNP combinations within the space of one minute. The identified SNP barcodes with the best performance between control and breast cancer groups were found to be control-dominant, suggesting that these SNP barcodes may prove protective against breast cancer. After statistical analysis, these control-dominant SNP barcodes were processed for odds ratio analysis for quantitative measurement with regard to the risk of breast cancer. CONCLUSION: This study proposes an effective high-speed method to analyze the SNP-SNP interactions for breast cancer association study. PMID- 19679064 TI - Apoptosis and cell cycle arrest of human colorectal cancer cell line HT-29 induced by vanillin. AB - BACKGROUND: Vanillin is responsible for the flavor and smell of vanilla, a widely used flavoring agent. Previous studies showed that vanillin could enhance the repair of mutations and thus function as an anti-mutagen. However, its role in cancer, a disease that is closely related to mutation has not yet been fully elucidated. METHODS: Hence, this study investigated the cytolytic and cytostatic properties of vanillin against HT-29, a human colorectal cancer cell line. Methods used including cell viability assay, acridine orange (AO)-ethidium bromide (EB) double staining cell morphological analysis, Cell cycle analysis, annexin V-propidium iodide apoptosis test and 5-bromo-2-deoxyuridine (BrdU) labeling cell proliferation assay. RESULTS: Results showed that apoptosis was induced by vanillin and the IC(50) for HT-29 and NIH/3T3 normal cell lines were 400 microg/ml and 1000 microg/ml, respectively. Different concentrations of vanillin arrest cell cycle at different checkpoints. 5-Bromo-2-deoxyuridine labeling cell proliferation assay showed that G0/G1 arrest was achieved at lower concentration of vanillin (200 microg/ml) while cell cycle analysis by flow cytometer showed that G2/M arrest occurs at higher concentration of vanillin (1000 microg/ml). CONCLUSION: Cytolytic and cytostatic effects shown by vanillin showed that it could be a useful colorectal cancer preventive agent. Further in vivo study should be carried out to confirm that similar effects could happen in animals. PMID- 19679065 TI - Clinical characteristics of tumors derived from colorectal cancer patients who harbor the tumor necrosis factor alpha-1031T/T and NOD2 3020insC polymorphism. AB - BACKGROUND: Genetic predispositions to disease have focused on highly penetrant causative changes in tumor suppressor genes or genes associated with DNA mismatch repair. New investigations are revealing new genetic associations with disease that are more subtle in their association with disease and require characterization. METHODS: In this report we have examined the tumor characteristics in a group of patients who have been shown to harbor two polymorphisms in two genes that are associated with the immune system NOD2 and TNFalpha. RESULTS: Colorectal cancers from patients with NOD2 3020insC and TNFalpha-1031T/T constitutional changes are mostly right-sided disease (OR=2.21, p=0.03) with a tendency to higher stages (OR=2.41, p=0.06), increased number of associated polyps (OR=1.77, p=0.16) and later age of average age of disease onset (p=0.039). CONCLUSION: The results reveal that there appear to be specific characteristics associated with the tumors that may aid in determining management strategies to reduce the risk of disease. PMID- 19679066 TI - IGF-1: elixir for motor neuron diseases. AB - Modulation of testosterone levels is a therapeutic approach for spinal and bulbar muscular atrophy (SBMA), a polyglutamine disorder that affects the motor neurons. The article by Palazzolo et al. in this issue of Neuron provides compelling evidence that the expression of insulin growth hormone is a potential therapeutic for SBMA. PMID- 19679067 TI - Too fat to fly? New brain circuits regulate obesity in Drosophila. AB - In mammals, fat store levels are regulated by brain centers that control food intake and metabolism. A new study by Al-Anzi and colleagues in this issue of Neuron identifies neurons with similar functions in Drosophila, further establishing the fly as a legitimate model to study obesity. PMID- 19679068 TI - The pre/post LTP debate. AB - The pre/post debate involves the question of whether long-term potentiation (LTP) is mediated by enhancement of release, enhancement of postsynaptic receptors, or both. Recent papers have presented evidence for purely postsynaptic or purely presynaptic changes, and a paper by Ahmed and Siegelbam (in this issue of Neuron) suggests a mechanism by which release is enhanced. This debate is increasingly constrained by technical advances that allow central synapses to be studied with increasing precision. A possible of way of reconciling conflicting evidence is suggested. PMID- 19679069 TI - A nonvisual look at the functional organization of visual cortex. AB - In this issue of Neuron, Mahon et al. show that the ventral visual cortex of congenitally blind individuals, who have never experienced the visual world, has an object-category organization similar to that found in sighted individuals. Here, we discuss the implications of this finding for our understanding of the "visual" cortex. PMID- 19679071 TI - Linking genetically defined neurons to behavior through a broadly applicable silencing allele. AB - Tools for suppressing synaptic transmission gain power when able to target highly selective neuron subtypes, thereby sharpening attainable links between neuron type, behavior, and disease; and when able to silence most any neuron subtype, thereby offering broad applicability. Here, we present such a tool, RC::PFtox, that harnesses breadth in scope along with high cell-type selection via combinatorial gene expression to deliver tetanus toxin light chain (tox), an inhibitor of vesicular neurotransmission. When applied in mice, we observed cell type-specific disruption of vesicle exocytosis accompanied by loss of excitatory postsynaptic currents and commensurately perturbed behaviors. Among various test populations, we applied RC::PFtox to silence serotonergic neurons, en masse or a subset defined combinatorially. Of the behavioral phenotypes observed upon en masse serotonergic silencing, only one mapped to the combinatorially defined subset. These findings provide evidence for separability by genetic lineage of serotonin-modulated behaviors; collectively, these findings demonstrate broad utility of RC::PFtox for dissecting neuron functions. PMID- 19679073 TI - Obesity-blocking neurons in Drosophila. AB - In mammals, fat store levels are communicated by leptin and insulin signaling to brain centers that regulate food intake and metabolism. By using transgenic manipulation of neural activity, we report the isolation of two distinct neuronal populations in flies that perform a similar function, the c673a-Gal4 and fruitless-Gal4 neurons. When either of these neuronal groups is silenced, fat store levels increase. This change is mediated through an increase in food intake and altered metabolism in c673a-Gal4-silenced flies, while silencing fruitless Gal4 neurons alters only metabolism. Hyperactivation of either neuronal group causes depletion of fat stores by increasing metabolic rate and decreasing fatty acid synthesis. Altering the activities of these neurons causes changes in expression of genes known to regulate fat utilization. Our results show that the fly brain measures fat store levels and can induce changes in food intake and metabolism to maintain them within normal limits. PMID- 19679070 TI - The role of apolipoprotein E in Alzheimer's disease. AB - The epsilon4 allele of apolipoprotein E (APOE) is the major genetic risk factor for Alzheimer's disease (AD). Although there have been numerous studies attempting to elucidate the underlying mechanism for this increased risk, how apoE4 influences AD onset and progression has yet to be proven. However, prevailing evidence suggests that the differential effects of apoE isoforms on Abeta aggregation and clearance play the major role in AD pathogenesis. Other potential mechanisms, such as the differential modulation of neurotoxicity and tau phosphorylation by apoE isoforms as well as its role in synaptic plasticity and neuroinflammation, have not been ruled out. Inconsistent results among studies have made it difficult to define whether the APOE epsilon4 allele represents a gain of toxic function, a loss of neuroprotective function, or both. Therapeutic strategies based on apoE propose to reduce the toxic effects of apoE4 or to restore the physiological, protective functions of apoE. PMID- 19679072 TI - Overexpression of IGF-1 in muscle attenuates disease in a mouse model of spinal and bulbar muscular atrophy. AB - Expansion of a polyglutamine tract in the androgen receptor (AR) causes spinal and bulbar muscular atrophy (SBMA). We previously showed that Akt-mediated phosphorylation of AR reduces ligand binding and attenuates the mutant AR toxicity. Here, we show that in culture insulin-like growth factor 1 (IGF-1) reduces AR aggregation and increases AR clearance via the ubiquitin-proteasome system through phosphorylation of AR by Akt. In vivo, SBMA transgenic mice overexpressing a muscle-specific isoform of IGF-1 selectively in skeletal muscle show evidence of increased Akt activation and AR phosphorylation and decreased AR aggregation. Augmentation of IGF-1/Akt signaling rescues behavioral and histopathological abnormalities, extends the life span, and reduces both muscle and spinal cord pathology of SBMA mice. This study establishes IGF-1/Akt-mediated inactivation of mutant AR as a strategy to counteract disease in vivo and demonstrates that skeletal muscle is a viable target tissue for therapeutic intervention in SBMA. PMID- 19679074 TI - Downregulation of NR3A-containing NMDARs is required for synapse maturation and memory consolidation. AB - NR3A is the only NMDA receptor (NMDAR) subunit that downregulates sharply prior to the onset of sensitive periods for plasticity, yet the functional importance of this transient expression remains unknown. To investigate whether removal/replacement of juvenile NR3A-containing NMDARs is involved in experience driven synapse maturation, we used a reversible transgenic system that prolonged NR3A expression in the forebrain. We found that removal of NR3A is required to develop strong NMDAR currents, full expression of long-term synaptic plasticity, a mature synaptic organization characterized by more synapses and larger postsynaptic densities, and the ability to form long-term memories. Deficits associated with prolonged NR3A were reversible, as late-onset suppression of transgene expression rescued both synaptic and memory impairments. Our results suggest that NR3A behaves as a molecular brake to prevent the premature strengthening and stabilization of excitatory synapses and that NR3A removal might thereby initiate critical stages of synapse maturation during early postnatal neural development. PMID- 19679075 TI - A role for SNAP25 in internalization of kainate receptors and synaptic plasticity. AB - Regulation of surface insertion and internalization of AMPA and NMDA receptors has emerged as a key mechanism for the control of synaptic strength. Regulatory elements for synaptic kainate receptors (KARs) are, however, largely undetermined. We have found that SNAP25 is critical for the synaptic removal of KARs, acting via GluK5 (i.e., KA2) subunits. SNAP25 coimmunoprecipitates with protein complexes containing PICK1, GRIP1, and GluK5 and colocalizes with GluK5 in both hippocampal neurons and transfected HEK293 cells. In hippocampal slices, purified SNAP25 antibodies and blocking peptides caused a GluK5-dependent run-up of KARs-mediated EPSC (EPSC(KAR)) recorded from CA3 pyramidal neurons when included in the patch pipette and prevented activity-dependent long-term depression of EPSC(KAR). As EPSC(KAR) LTD, SNAP25/PICK1/GluK5 interactions are dynamically regulated by PKC. PMID- 19679076 TI - Recruitment of N-Type Ca(2+) channels during LTP enhances low release efficacy of hippocampal CA1 perforant path synapses. AB - The entorhinal cortex provides both direct and indirect inputs to hippocampal CA1 neurons through the perforant path and Schaffer collateral synapses, respectively. Using both two-photon imaging of synaptic vesicle cycling and electrophysiological recordings, we found that the efficacy of transmitter release at perforant path synapses is lower than at Schaffer collateral inputs. This difference is due to the greater contribution to release by presynaptic N type voltage-gated Ca(2+) channels at the Schaffer collateral than perforant path synapses. Induction of long-term potentiation that depends on activation of NMDA receptors and L-type voltage-gated Ca(2+) channels enhances the low efficacy of release at perforant path synapses by increasing the contribution of N-type channels to exocytosis. This represents a previously uncharacterized presynaptic mechanism for fine-tuning release properties of distinct classes of synapses onto a common postsynaptic neuron and for regulating synaptic function during long term synaptic plasticity. PMID- 19679077 TI - Serial, covert shifts of attention during visual search are reflected by the frontal eye fields and correlated with population oscillations. AB - Attention regulates the flood of sensory information into a manageable stream, and so understanding how attention is controlled is central to understanding cognition. Competing theories suggest visual search involves serial and/or parallel allocation of attention, but there is little direct, neural evidence for either mechanism. Two monkeys were trained to covertly search an array for a target stimulus under visual search (endogenous) and pop-out (exogenous) conditions. Here, we present neural evidence in the frontal eye fields (FEF) for serial, covert shifts of attention during search but not pop-out. Furthermore, attention shifts reflected in FEF spiking activity were correlated with 18-34 Hz oscillations in the local field potential, suggesting a "clocking" signal. This provides direct neural evidence that primates can spontaneously adopt a serial search strategy and that these serial covert shifts of attention are directed by the FEF. It also suggests that neuron population oscillations may regulate the timing of cognitive processing. PMID- 19679079 TI - Motor learning is optimally tuned to the properties of motor noise. AB - In motor learning, our brain uses movement errors to adjust planning of future movements. This process has traditionally been studied by examining how motor planning is adjusted in response to visuomotor or dynamic perturbations. Here, I show that the learning strategy can be better identified from the statistics of movements made in the absence of perturbations. The strategy identified this way differs from the learning mechanism assumed in mainstream models for motor learning. Crucial for this strategy is that motor noise arises partly centrally, in movement planning, and partly peripherally, in movement execution. Corrections are made by modification of central planning signals from the previous movement, which include the effects of planning but not execution noise. The size of the corrections is such that the movement variability is minimized. This physiologically plausible strategy is optimally tuned to the properties of motor noise, and likely underlies learning in many motor tasks. PMID- 19679078 TI - Category-specific organization in the human brain does not require visual experience. AB - Distinct regions within the ventral visual pathway show neural specialization for nonliving and living stimuli (e.g., tools, houses versus animals, faces). The causes of these category preferences are widely debated. Using functional magnetic resonance imaging, we find that the same regions of the ventral stream that show category preferences for nonliving stimuli and animals in sighted adults show the same category preferences in adults who are blind since birth. Both blind and sighted participants had larger blood oxygen-level dependent (BOLD) responses in the medial fusiform gyrus for nonliving stimuli compared to animal stimuli and differential BOLD responses in lateral occipital cortex for animal stimuli compared to nonliving stimuli. These findings demonstrate that the medial-to-lateral bias by conceptual domain in the ventral visual pathway does not require visual experience in order to develop and suggest the operation of innately determined domain-specific constraints on the organization of object knowledge. PMID- 19679080 TI - How do adhesion proteins stick? AB - Cadherin homodimerization mediates cell-cell adhesion, but one stabilizing structural element has inspired questions about assembly mechanisms. Combining single molecule fluorescence and force analyses allowed Sivasankar et al. to provide strong evidence for an induced fit pathway to dimerization. PMID- 19679081 TI - Nup120: one more piece in the NPC puzzle. AB - In this issue of Structure, Schwartz and coworkers present the structure of Nup120, a nucleoporin of the nuclear pore scaffold. The structure shows that, in contrast to earlier predictions, the nucleoporins have a larger fold repertoire than expected. PMID- 19679082 TI - Catalytically incompetent by design. AB - Sondermann and colleagues have characterized FimX, a protein with degenerate GGDEF and EAL domains. The study confirms the expected domain folds lacking conserved catalytic residues for c-di-GMP synthesis/degradation, and also defines domain arrangements, providing insight to regulatory mechanisms. PMID- 19679083 TI - Out of the box binding determines specificity of SH2 domain interaction. AB - SH2 domains are phosphotyrosine specific interaction modules with largely overlapping sequence specificities. A recent structure by Bae et al. revealed that SH2 domain specificity can be mediated by secondary binding sites located outside the phosphotyrosine binding pocket. PMID- 19679084 TI - The origin of allosteric functional modulation: multiple pre-existing pathways. AB - Although allostery draws increasing attention, not much is known about allosteric mechanisms. Here we argue that in all proteins, allosteric signals transmit through multiple, pre-existing pathways; which pathways dominate depend on protein topologies, specific binding events, covalent modifications, and cellular (environmental) conditions. Further, perturbation events at any site on the protein surface (or in the interior) will not create new pathways but only shift the pre-existing ensemble of pathways. Drugs binding at different sites or mutational events in disease shift the ensemble toward the same conformations; however, the relative populations of the different states will change. Consequently the observed functional, conformational, and dynamic effects will be different. This is the origin of allosteric functional modulation in dynamic proteins: allostery does not necessarily need to invoke conformational rearrangements to control protein activity and pre-existing pathways are always defaulted to during allostery regardless of the stimulant and perturbation site in the protein. PMID- 19679085 TI - The CATH hierarchy revisited-structural divergence in domain superfamilies and the continuity of fold space. AB - This paper explores the structural continuum in CATH and the extent to which superfamilies adopt distinct folds. Although most superfamilies are structurally conserved, in some of the most highly populated superfamilies (4% of all superfamilies) there is considerable structural divergence. While relatives share a similar fold in the evolutionary conserved core, diverse elaborations to this core can result in significant differences in the global structures. Applying similar protocols to examine the extent to which structural overlaps occur between different fold groups, it appears this effect is confined to just a few architectures and is largely due to small, recurring super-secondary motifs (e.g., alphabeta-motifs, alpha-hairpins). Although 24% of superfamilies overlap with superfamilies having different folds, only 14% of nonredundant structures in CATH are involved in overlaps. Nevertheless, the existence of these overlaps suggests that, in some regions of structure space, the fold universe should be seen as more continuous. PMID- 19679086 TI - Multimeric options for the auto-activation of the Saccharomyces cerevisiae FAS type I megasynthase. AB - The fungal type I fatty acid synthase (FAS) is a 2.6 MDa multienzyme complex, catalyzing all necessary steps for the synthesis of long acyl chains. To be catalytically competent, the FAS must be activated by a posttranslational modification of the central acyl carrier domain (ACP) by an intrinsic phosphopantetheine transferase (PPT). However, recent X-ray structures of the fungal FAS revealed a barrel-shaped architecture, with PPT located at the outside of the barrel wall, spatially separated from the ACP caged in the inner volume. This separation indicated that the activation has to proceed before the assembly to the mature complex, in a conformation where the ACP and PPT domains can meet. To gain insight into the auto-activation reaction and also into the fungal FAS assembly pathway, we structurally and functionally characterized the Saccharomyces cerevisiae FAS type I PPT as part of the multienzyme protein and as an isolated domain. PMID- 19679087 TI - Structural determinants of transmembrane helical proteins. AB - We identify a structural feature of transmembrane helical proteins that restricts their conformational space and suggests a new way of understanding the construction and stability of their native states. We show that five kinds of well-known specific favorable interhelical interactions (hydrogen bonds, aromatic interactions, salt bridges, and two interactions from packing motifs) precisely determine the packing of the transmembrane helices in 15 diverse proteins. To show this, we iteratively reassemble the helix bundle of each protein using only these interactions, generic interaction geometries, and individual helix backbone conformations. On average, the representative set of rebuilt structures best satisfying the constraints imposed by the five types of interhelical interactions has an average Calpha root-mean-square deviation from the native of 1.03 A. Implications for protein folding, structure and motion prediction, modeling, and design are discussed. PMID- 19679089 TI - A surface loop directs conformational switching of a lipoyl domain between a folded and a novel misfolded structure. AB - A prominent surface loop links the first two beta strands of the lipoyl domain (E2plip) from the pyruvate dehydrogenase multienzyme complex of Escherichia coli. We show here that shortening this loop by two residues generates a protein that populates two structurally distinct stable conformers: an active, native-like monomer (HM) and a functionally compromised misfolded dimer (LM). Conversion of LM to HM was observed after exposure to temperatures above 50 degrees C. Removal of two additional residues from the loop caused the protein to adopt exclusively the misfolded conformation. Detailed NMR structural studies of the misfolded dimer reveal that the N-terminal half of the domain was unfolded and dynamic, whereas the C-terminal halves of two monomers had associated to form a structure with two-fold symmetry and a topology mimicking that of the folded monomer. The surface loop is therefore a hitherto unsuspected determinant in the folding process that leads to a functional protein. PMID- 19679088 TI - Structural analysis of the GGDEF-EAL domain-containing c-di-GMP receptor FimX. AB - Bacterial pathogenesis involves social behavior including biofilm formation and swarming, processes that are regulated by the bacterially unique second messenger cyclic di-GMP (c-di-GMP). Diguanylate cyclases containing GGDEF and phosphodiesterases containing EAL domains have been identified as the enzymes controlling cellular c-di-GMP levels, yet less is known regarding signal transmission and the targets of c-di-GMP. FimX, a protein from Pseudomonas aeruginosa that governs twitching motility, belongs to a large subfamily containing both GGDEF and EAL domains. Biochemical and structural analyses reveals its function as a high-affinity receptor for c-di-GMP. A model for full length FimX was generated combining solution scattering data and crystal structures of the degenerate GGDEF and EAL domains. Although FimX forms a dimer in solution via the N-terminal domains, a crystallographic EAL domain dimer suggests modes for the regulation of FimX by c-di-GMP binding. The results provide the structural basis for c-di-GMP sensing via degenerate phosphodiesterases. PMID- 19679090 TI - Protein-peptide interactions adopt the same structural motifs as monomeric protein folds. AB - We compared the modes of interaction between protein-peptide interfaces and those observed within monomeric proteins and found surprisingly few differences. Over 65% of 731 protein-peptide interfaces could be reconstructed within 1 A RMSD using solely fragment interactions occurring in monomeric proteins. Interestingly, more than 80% of interacting fragments used in reconstructing a protein-peptide binding site were obtained from monomeric proteins of an entirely different structural classification, with an average sequence identity below 15%. Nevertheless, geometric properties perfectly match the interaction patterns observed within monomeric proteins. We show the usefulness of our approach by redesigning the interaction scaffold of nine protein-peptide complexes, for which five of the peptides can be modeled within 1 A RMSD of the original peptide position. These data suggest that the wealth of structural data on monomeric proteins could be harvested to model protein-peptide interactions and, more importantly, that sequence homology is no prerequisite. PMID- 19679092 TI - Epidemiology of human leptospirosis in Trinidad and Tobago, 1996-2007: a retrospective study. AB - A retrospective study to describe the epidemiology of human leptospirosis in Trinidad and Tobago during 1996-2007 was conducted. All confirmed cases of leptospirosis was analyzed according to age, sex, seasonality and geographic distribution. A total of 278 cases were recorded, with an average annual incidence rate of 1.84 per 100,000 population. Seventy-five percent of the cases occurred during the wet season, with the highest number of cases recorded in November. A positive correlation was found between number of cases and rainfall. Males constituted 80% of all cases, and the overall male:female ratio was 4.6:1. The total case fatality rate was 5.8%, with deaths among males four times more common than in females. Clinical leptospirosis was greatest in the 10-19 age group and lowest in the 0-9 age group. The total prevalence was 22 per 100,000 population, with the highest prevalence 41 per 100,000 recorded in the regional corporation of Sangre Grande and the lowest (6 per 100,000) in the city of Port of Spain. The lack of important information and active surveillance showed that the level of awareness of the disease is low in the country. The disease is still under-reported, and is considered to be of significant public health importance. PMID- 19679091 TI - A tetrahedral transition state at the active sites of the 20S proteasome is coupled to opening of the alpha-ring channel. AB - Intrinsic conformational transitions contribute to the catalytic action of many enzymes. Here we use a single-molecule approach to demonstrate how such transitions are linked to the catalytic sites of the eukaryotic proteasome, an essential protease of the ubiquitin pathway. The active sites of the cylindrical proteasomal core particle are located in a central chamber accessible through gated entry channels. By using atomic force microscopy, we found continual alternation between open and closed gate conformations. We analyzed the relative abundance of these conformers in wild-type and mutated yeast core particles upon exposure to substrates or inhibitors. Our data indicate that the dynamic gate can be opened by allosteric coupling to a tetrahedral transition state at any of the working active centers. The results point to the N(alpha)-amine of the N-terminal active site threonyl residue as the major effector group responsible for triggering the essential conformational switch. PMID- 19679093 TI - Species B of Anopheles culicifacies (Diptera: Culicidae) is reproductively less fit than species A and C of the complex. AB - Anopheles culicifacies, the most important malaria vector of peninsular India exist as a complex of five sibling species. The member species of the complex have various biological differences including their susceptibility to malaria parasites. The present attempt is made to study and compare the fecundity of the differentially susceptible members of the An. culicifacies complex. Gravid female mosquitoes of species A, B and C were allowed to lay their eggs individually during first and second gonotrophic cycle. The eggs were counted after hatching and categorized as 'hatched eggs', 'unhatched eggs', 'embryonated eggs', 'unembryonated eggs' and 'non/partially melanized eggs'. The data was analyzed using Student's t test, ANOVA, Chi-square and Pearson's correlation analysis. All females that were visually categorized as 'gravid' did not lay eggs. Species C laid maximum number of eggs per female. The eggs laid per female mosquito of each species were found to be significantly higher during second gonotrophic cycle as compared with the first gonotrophic cycle. The eggs hatched per female in species C were found to be significantly higher than that of species A and B. The poor vector species B mosquitoes were found to be the least fecund among the members of the species complex. The unembryonated eggs constitute the biggest proportion of the unhatched eggs in species A, B and C of the species complex. PMID- 19679094 TI - Peeling as a novel, simple, and effective method for isolation of apical membrane from intact polarized epithelial cells. AB - Apical membrane of polarized epithelial cells is generally isolated by physicochemical methods, that is, precipitation with polyethylene glycol (PEG) or MgCl(2) followed by differential centrifugation or sucrose density gradient centrifugation. However, these protocols are considerably sophisticated and frequently accompanied by impurities (e.g., contaminations of basolateral membrane and intracellular organelles), particularly by inexperienced investigators. We have developed a simple and effective method for isolation of apical membrane from intact polarized renal tubular epithelial cells. On the basis of hydrous affinity and/or ionic interaction, the apical membrane could be efficiently peeled from the cells by four different materials-Whatman filter paper, nitrocellulose membrane, cellophane, and glass coverslip-all of which are available in most research laboratories. Phase-contrast and laser-scanning confocal microscopic examinations using anti-ZO-1 antibody showed that other parts of the cells, particularly tight junction complex, remained intact after peeling by all four of these surfaces. Western blot analyses of gp135 (apical membrane marker) and of Na(+)/K(+)-ATPase, LAMP-2, COX-4, and calpain-1 (markers of basolateral membrane, lysosome, mitochondria, and cytosolic compartment, respectively) revealed that peeling with Whatman filter paper and glass coverslip was most and second-most effective, respectively, without any contaminations from basolateral membrane and other intracellular organelles that could be detected in the samples isolated by peeling with nitrocellulose membrane and cellophane and by conventional methods (i.e., precipitation with PEG or MgCl(2) followed by differential centrifugation or sucrose density gradient centrifugation). Our physical method is very simple, easy to follow (even by inexperienced investigators), time-saving, and cost-effective with a higher efficiency (as compared with conventional methods) for isolation of apical membrane from polarized epithelial cells. PMID- 19679095 TI - Measurement of the molecular masses of hydrophilic and hydrophobic subunits of ATP synthase and complex I in a single experiment. AB - The adenosine triphosphate (ATP) synthase and complex I in mitochondria are membrane-bound multisubunit assemblies of both hydrophilic and hydrophobic proteins. Hitherto, the mass spectrometric measurement of their molecular masses has required that many of the hydrophobic proteins be analyzed separately from the other components in two different experiments. Here we describe a procedure that allows the molecular masses of all, or nearly all, of the subunits of each complex to be measured in a single experiment. The key feature is a mobile phase, in which hydrophilic and hydrophobic components remain soluble, that is compatible with reverse phase chromatography. In this way, the masses of all 17 subunits of bovine ATP synthase, 14 of the 17 subunits of the enzyme from Saccharomyces cerevisiae, 42 of the 45 subunits of bovine complex I, and all 28 of the subunits of bovine subcomplex Ialpha were measured. The method was used to characterize the subunits of ATP synthases and complexes I from a variety of species and to follow the progress of mild trypsinolysis of ATP synthase. It could be applied to other respiratory and photosynthetic complexes and, in general, to any protein complex that contains both hydrophilic and hydrophobic subunits. PMID- 19679096 TI - Quantitative label-free screening for antibodies using scattering biophotonic microarray imaging. AB - A biophotonic array based on gold nanoparticles functionalized with antigen proteins has been used to determine the concentrations of the respective antibodies in solution. Four proteins-fibrinogen, bovine serum albumin, transferrin, and C-reactive protein-were used to construct a test array with the assay repeated a number of times. The antibody-antigen association and dissociation rate constants were determined for the antibody assays from a series of calibration experiments. The label-free determination of the unknown antibody concentrations was performed using two related kinetic analyses. From these results, the current array assay sensitivity is 250 ng ml(-1) with an accuracy of 15% using an 8-min kinetic measurement and a 16-spot averaged assay. PMID- 19679097 TI - Flow cytometry-based assay for the activity of NAD(P)H oxidoreductases of the outer mitochondrial membrane. AB - NAD(P)H oxidoreductases of the outer mitochondrial membrane (OMM) are able to activate various xenobiotics and stimulate the production of reactive oxygen species and the opening of the mitochondrial permeability transition pore. However, the role of these systems in the cell damage by xenobiotics and chemotherapeutic drugs is poorly understood because the methods for the selective assessment of their activity have not been elaborated and specific inhibitors are unknown. Here we propose a method for the semiquantitative assessment of the activity of NAD(P)H oxidoreductases of the OMM in intact and permeabilized cells that is based on the flow cytometry detection of dimethylbiacridene, a fluorescent product of two-electron reduction of lucigenin. The method uses the structural feature of mitochondrial organization: the proximity of the sites of one-electron reduction of lucigenin to cation radical (NAD(P)H oxidoreductases of the OMM) to the sites of its subsequent oxidation (cytochrome c oxidase). The inhibition of cytochrome c oxidase by cyanide selectively activates the dimethylbiacridene formation by oxidoreductases of the OMM but not by other cellular oxidoreductases. The proposed protocol allows one to assess the lucigenin reductase (two-electron) activity of NAD(P)H oxidoreductases of the OMM and to compare it with the activity of other cellular systems that can be used for the analysis of the role of these systems in the cell damage by xenobiotics and antitumor drugs. PMID- 19679098 TI - Supercomplexes of the mitochondrial electron transport chain decline in the aging rat heart. AB - Accumulation of mitochondrial electron transport chain (ETC) defects is a recognized hallmark of the age-associated decline in cardiac bioenergetics; however, the molecular events involved are only poorly understood. In the present work, we hypothesized that age-related ETC deterioration stemmed partly from disassociation of large solid-state macromolecular assemblies termed "supercomplexes". Mitochondrial proteins from young and old rat hearts were separated by blue native-PAGE, protein bands analyzed by LC-MALDI-MS/MS, and protein levels quantified by densitometry. Results showed that supercomplexes comprised of various stoichiometries of complexes I, III and IV were observed, and declined significantly (p<0.05, n=4) with age. Supercomplexes displaying the highest molecular masses were the most severely affected. Considering that certain diseases (e.g. Barth Syndrome) display similar supercomplex destabilization as our results for aging, the deterioration in ETC supercomplexes may be an important underlying factor for both impaired mitochondrial function and loss of cardiac bioenergetics with age. PMID- 19679099 TI - Systematic gene regulation involving miRNAs during neuronal differentiation of mouse P19 embryonic carcinoma cell. AB - MicroRNAs (miRNAs) are small noncoding RNA and play an essential role in gene regulation. In this study, we investigated regulation of gene expression during neuronal differentiation of mouse P19 embryonic carcinoma cells and described a systematic pathway of gene regulation involving miRNAs. In the pathway, downregulation of Lin28 involved in blocking the let-7 maturation and upregulation of let-7 occur following induction of the differentiation, thereby triggering suppression of the downstream High Mobility Group A2 (Hmga2) gene expression via activation of gene silencing mediated by let-7. Our data further suggest that miR-9, as well as miR-125b, participate in the reduction of the Lin28 expression. The gene regulation involving miRNAs likely contributes to a rapid and programmed change in gene expression in neuronal differentiation of P19 cells. PMID- 19679101 TI - Lesser known relatives of miRNA. AB - Since the discovery of RNAi (RNA interference) major attention has focused on studying miRNA (microRNA) and siRNA (small interfering RNA). However, within the last few years, several other small ncRNAs (non-coding RNAs) have been discovered and thus various newer acronyms representing these 'other' classes of small ncRNAs have populated the literature. Of these, piRNA (Piwi-interacting RNA) has been gaining importance because of its role as the guardian of the germline genome. Some of the other newly discovered small ncRNAs have been mostly reported in plants, and they are yet to be studied more comprehensively. Nevertheless, piRNA and the 'other' small ncRNAs deserve some discussion because they are members of the increasingly large family of small ncRNAs. PMID- 19679100 TI - Articular cartilage vesicles contain RNA. AB - Small membrane-bound extracellular organelles known as articular cartilage matrix vesicles (ACVs) participate in pathologic mineralization in osteoarthritic articular cartilage. ACVs are also present in normal cartilage, although they have no known functions other than mineralization. Recently, RNA was identified in extracellular vesicles derived from mast cells, suggesting that such vesicles might carry coding information from cell to cell. We found that ACVs from normal porcine and human articular cartilage and primary chondrocyte conditioned media contained 1 microg RNA/80 microg ACV protein. No DNA could be detected. RT-PCR of ACV RNA demonstrated the presence of full length mRNAs for factor XIIIA, type II transglutaminase, collagen II, aggrecan, ANKH and GAPDH. RNA in intact ACVs was resistant to RNase, despite the fact that ACV preparations contained measurable levels of active RNases. Significantly, radiolabeled RNA in ACVs could be transferred to unlabeled chondrocytes by co-incubation and produced changes in levels of chondrocyte enzymes and proteins. The demonstration that ACVs contain mRNAs suggests that they may function to shuttle genetic information between articular cells and indicate novel functions for these structures in articular cartilage. PMID- 19679102 TI - Anandamide increases swelling and reduces calcium sensitivity of mitochondria. AB - The endocannabinoid anandamide alters mitochondria-dependent signal transduction, thus controlling key cellular events like energy homeostasis and induction of apoptosis. Here, the ability of anandamide to directly affect the integrity of mitochondria was investigated on isolated organelles. We found that anandamide dose-dependently increases mitochondrial swelling, and reduces cytochrome c release induced by calcium ions. The effects of anandamide were independent of its target receptors (e.g., cannabinoid or vanilloid receptors), and were paralleled by decreased membrane potential and increased membrane fluidity. Overall, our data suggest that anandamide can impact mitochondrial physiology, by reducing calcium sensitivity and perturbing membrane properties of these organelles. PMID- 19679103 TI - LRRC19, a novel member of the leucine-rich repeat protein family, activates NF kappaB and induces expression of proinflammatory cytokines. AB - We have identified a new functional transmembrane receptor, LRRC19 (leucine-rich repeat containing 19), that belongs to the LRR protein family. LRRC19's central core has four analogous LRR repeating modules in a juxtaposed array and a casein kinase (CK2) phosphorylation site in the cytoplasmic domain. LRRC19 mRNA was found in the kidney, spleen and intestine of adult mice using both RT-PCR and in situ hybridization. LRRC19 does not contain a cytoplasmic Toll/IL-1 receptor (TIR) domain but was able to activate NF-kappaB and induce production of proinflammatory cytokines. LRRC19 shares a close evolutionary relationship with multiple Toll-like receptors (TLRs), especially TLR3. Importantly, the TLR3 ligand, as well as other TLR ligands, significantly promoted the expression of proinflammatory cytokines and the activation of NF-kappaB by LRRC19. Thus, LRRC19 may play an important role in inducing innate immune responses in certain tissues such as the kidney. PMID- 19679104 TI - Dietary antioxidants interfere with Amplex Red-coupled-fluorescence assays. AB - Oxidation of Amplex Red by hydrogen peroxide in the presence of horseradish peroxidase (HRP) gives rise to an intensely colour product, resorufin. This reaction has been frequently employed for measurements based on enzyme-coupled reactions that detect hydrogen peroxide as a final reaction product. In the current study, we show that the presence of dietary antioxidants at biological concentrations in the reaction medium produced interferences in the Amplex Red/HRP catalyzed reaction that result in an over quantification of the hydrogen peroxide produced. The interference observed showed a dose-dependent manner, and a possible mechanism of interaction of dietary antioxidants with HRP in the Amplex Red-coupled-fluorescent assay is proposed. PMID- 19679105 TI - Mechanism of cytotoxicity mediated by the C31 fragment of the amyloid precursor protein. AB - The cytoplasmic tail of the amyloid precursor protein (APP) contains two putatively cytotoxic peptides, Jcasp and C31, derived by caspase cleavage of APP. Jcasp is a fragment starting from the epsilon-secretase site to position 664, while C31 is a fragment from position 665 to the C-terminus. Our studies now showed that compared to C31, Jcasp appeared to play a minor role in cytotoxicity. In particular, inhibition of Jcasp generation by treatment of gamma-secretase inhibitor did not lead to any attenuation of C31-induced toxicity. Secondly, because C31 toxicity is largely absent in cells lacking endogenous APP, we determined, using a split beta-galactosidase complementary assay to monitor protein-protein interactions, the presence of APP associated complexes. Our results demonstrated that both APP homomeric and C31/APP heteromeric complexes were correlated with cell death, indicating that C31 complexes with APP to recruit the interacting partners that initiate the signals related to cellular toxicity. PMID- 19679106 TI - TCP-FA4: a derivative of tranylcypromine showing improved blood-brain permeability. AB - A variety of approaches have been taken to improve the brain penetration of pharmaceutical agents. The amphipathic character of a compound can improve its interaction with the lipid bilayer within cell membranes, and as a result improve permeability. Fatty acid chains or lipoamino acids of various lengths were attached to tranylcypromine (TCP), in an attempt to improve the blood-brain barrier (BBB) permeability by increasing the lipophilicity as well as the amphiphatic character of the drug. TCP-FA4, one of the derivatives containing a four carbon alkyl acid chain, showed the greatest improvement in permeability. This molecule was slightly neuroprotective in a beta-amyloid-induced neurodegeneration assay and may also be capable of upregulating brain derived neurotrophic factor (BDNF), as indicated by cell culture assays using human umbilical vein endothelial cells. Since decreased levels of BDNF are observed in many CNS disorders, and direct injection of BDNF is not a viable option due to its poor permeability across the BBB, small molecules capable of regulating BDNF that also cross the BBB may be an interesting treatment option. PMID- 19679107 TI - Metabolism and biological production of resolvins derived from docosapentaenoic acid (DPAn-6). AB - 17S-HDPAn-6 (17S-hydroxydocosa-4Z,7Z,10Z,13Z,15E-pentaenoic acid) and 10S,17S HDPAn-6 (10S,17S-dihydroxydocosa-4Z,7Z,11E,13Z,15E-pentaenoic acid) are potent anti-inflammatory resolvins derived from DPAn-6 (docosapentaenoic acid n-6) and are analogous in structure and action to DHA (docosahexaenoic acid)-derived resolvins. These resolvins have proven to be potential drug candidates, albeit with therapeutic profiles that need optimization. The main objectives of this study were to evaluate key features of DPAn-6 derived resolvins that are important for therapeutic efficacy, demonstrate that these DPAn-6 resolvins could be produced naturally, and could therefore have physiological significance. Here we demonstrate biological production, examine pharmacokinetic profiles and identify key routes of metabolic inactivation of DPAn-6 derived resolvins. We compare their metabolic stability to a known resolvin, 17S-HDHA (17S hydroxydocosa-4Z,7Z,10Z,13Z,15E,19Z-hexaenoic acid) and show that order of their stabilities is 10S,17S-HDPAn-6>17S-HDPAn-6>17S-HDHA. We show that both these compounds are not strong inhibitors of cytochrome-P450 enzymes. We evaluate activity of compounds in the delayed-type hypersensitivity model, results of which show that compounds need optimization for enhanced duration and magnitude of action. Analysis of the metabolic stability and identification of metabolites of these compounds could play an important role in the design of better analogs with longer durations of action and hence better efficacy. PMID- 19679108 TI - Identification of the major human hepatic and placental enzymes responsible for the biotransformation of glyburide. AB - One of the factors affecting the pharmacokinetics (PK) of a drug during pregnancy is the activity of hepatic and placental metabolizing enzymes. Recently, we reported on the biotransformation of glyburide by human hepatic and placental microsomes to six metabolites that are structurally identical between the two tissues. Two of the metabolites, 4-trans-(M1) and 3-cis-hydroxycyclohexyl glyburide (M2b), were previously identified in plasma and urine of patients treated with glyburide and are pharmacologically active. The aim of this investigation was to identify the major human hepatic and placental CYP450 isozymes responsible for the formation of each metabolite of glyburide. This was achieved by the use of chemical inhibitors selective for individual CYP isozymes and antibodies raised against them. The identification was confirmed by the kinetic constants for the biotransformation of glyburide by cDNA-expressed enzymes. The data revealed that the major hepatic isozymes responsible for the formation of each metabolite are as follows: CYP3A4 (ethylene-hydroxylated glyburide (M5), 3-trans-(M3) and 2-trans-(M4) cyclohexyl glyburide); CYP2C9 (M1, M2a (4-cis-) and M2b); CYP2C8 (M1 and M2b); and CYP2C19 (M2a). Human placental microsomal CYP19/aromatase was the major isozyme responsible for the biotransformation of glyburide to predominantly M5. The formation of significant amounts of M5 by CYP19 in the placenta could render this metabolite more accessible to the fetal circulation. The multiplicity of enzymes biotransforming glyburide and the metabolites formed underscores the potential for its drug interactions in vivo. PMID- 19679109 TI - Beneficial actions of oleanolic acid in an experimental model of multiple sclerosis: a potential therapeutic role. AB - Multiple sclerosis (MS) is a chronic autoimmune inflammatory disease for which there exist no therapies without undesired side effects. Thus, the establishment of less toxic treatments is an ongoing challenge. Nowadays, research on medicinal plants has been attracting much attention, since screening of its active principles could prove useful in identification of safe and innovative pharmaceutical molecules. In this study we investigated the therapeutic effect of oleanolic acid (OA) a plant-derived triterpene with potent anti-inflammatory and immunomodulatory activities, whose actions on CNS diseases remain far from completely characterized. We focussed on the potential therapeutic effect of oleanolic acid (OA) on an accepted experimental model of MS, the experimental autoimmune encephalomyelitis (EAE). We have found that OA treatment, before or at the early onset of EAE, ameliorates neurological signs of EAE-mice. These beneficial effects of OA seem to be associated with a reduction of blood-brain barrier leakage and lower infiltration of inflammatory cells within the CNS, as well as with its modulatory role in Th1/Th2 polarization: inhibition of proinflammatory cytokines and chemokines, and stimulation of anti-inflammatory ones. Moreover, EAE-animals that were treated with OA had lower levels of anti MOG antibodies than untreated EAE-mice. Our findings show that the administration of the natural triterpenoid OA reduces and limits the severity and development of EAE. Therefore, OA therapy might be of clinical interest for human MS and other Th1 cell-mediated inflammatory diseases. PMID- 19679110 TI - Insulin-stimulated translocation of GLUT4 to the plasma membrane in rat hippocampus is PI3-kinase dependent. AB - In the central nervous system (CNS) insulin mediates a variety of effects including feeding, metabolism and cognition. The cognitive enhancing effects of insulin are proposed to be mediated through activation of insulin receptors in the hippocampus, an important integration center for learning and memory in the mammalian brain. Since less is known regarding insulin signaling events in the hippocampus, the aim of the current study was to determine whether insulin stimulates similar signaling cascades and GLUT4 translocation in the rat hippocampus as has been described in peripheral tissues. Intracerebroventricular administration of insulin increases hippocampal insulin levels and also stimulates the phosphorylation of Akt in a time-dependent manner. Insulin also stimulates the translocation of GLUT4 to hippocampal plasma membranes in a time course that mirrors the increases in glucose uptake observed during the performance of hippocampal-dependent tasks. Insulin stimulated phosphorylation of Akt and translocation of GLUT4 were blocked by pretreatment with the PI3-kinase inhibitor LY294002. Confocal immunofluorescence determined that insulin stimulated phosphorylation of Akt was localized to neurons and colocalized with the insulin receptor and GLUT4 in the rat hippocampus, thereby identifying the functional anatomical substrates of insulin signaling in the hippocampus. These results demonstrate that insulin-stimulated translocation of GLUT4 to the plasma membrane in the rat hippocampus occurs via similar mechanisms as described in peripheral tissues and suggests that insulin-mediated translocation of GLUT4 may provide a mechanism through which hippocampal neurons rapidly increase glucose utilization during increases in neuronal activity associated with hippocampal dependent learning. PMID- 19679111 TI - Forty-hertz triple-pulse stimulation induces motor cortical facilitation in humans. AB - A single pulse of transcranial magnetic stimulation (TMS) can reset the 15- to 30 Hz beta-band oscillations in the motor cortex. These oscillations are known to influence the amplitude of corticospinal activity evoked by TMS. To garner further evidence for this resetting, we tested how electromyographic responses to motor cortex TMS were modulated by a preceding series of TMS pulses. We used a triad of conditioning TMS pulses at various interstimulus intervals (ISIs) in an attempt to drive cortical activity at the corresponding frequency. We then analyzed how the amplitude of motor-evoked potentials (MEPs) to a test pulse varied at different intervals after the conditioning triad. When conditioning pulses were given at an ISI of 25 ms, responses to the fourth (test) pulse were facilitated 25 ms later. Neither a single conditioning pulse nor triad of conditioning pulses separated by other ISIs enhanced responses to the test pulse at the expected timings. Triads of pulses at an ISI of 25 ms did not enhance subsequent MEPs to brainstem stimulation. Based on the intensity of the conditioning stimuli necessary to produce this effect and on the effective interval, we conclude that the facilitation at 25 ms differs from intracortical facilitation at 7-10 ms seen in the paired-pulse experiment originally reported by Kujirai et al. These results suggest that a triad of TMS pulses can enhance an intrinsic oscillatory rhythm of the motor cortex (40 Hz) and facilitate cortical activity at an ISI corresponding to the frequency of that rhythm. PMID- 19679112 TI - Amygdala responses to positively and negatively valenced baby faces in healthy female volunteers: influences of individual differences in harm avoidance. AB - Research regarding the lateralization of processing emotional visual stimuli suggests various roles for affective information-processing by the amygdalae. However, individual differences seem to influence outcome results. In this study we re-investigate this question, paying special attention to the salient nature of the mood inducing stimuli. We presented blocks of happy looking baby faces and sad looking baby faces (disfigured by severe dermatological conditions), as well as blurred isoluminescent neutral pictures to a 'homogeneous' group of 40 healthy female subjects during fMRI. We used the temperament dimension harm avoidance (HA), extracted from the Temperament and Character Inventory (TCI), to evaluate the impact of this personality feature on 'emotional' amygdala responses. HA is related to behavioral inhibition and it implies a genetically determined bias towards being cautious, apprehensive and overly pessimistic. Because emotional valence and arousal may be controlled by different neural systems, the positively and negatively valenced baby faces were selected to be equal in arousal levels. Viewing blocks of negatively valenced baby faces evoked bilateral amygdala activity, whereas viewing positively valenced ones resulted in left amygdala activity only. Globally, we found no evidence of lateralized amygdala specialization. When taking into account individual differences in HA, only in female subjects who score high on this dimension did we find predominantly left amygdala activation when viewing blocks of negatively valenced baby faces. HA did not influence amygdala activity when processing positively valenced images. Our results might indicate that personality features, such as HA, could be of importance in 'emotional' fMRI paradigms examining amygdala lateralization patterns. PMID- 19679114 TI - Speeding before and slowing after errors: is it all just strategy? AB - People are usually faster before and slower after committing an error. This finding has traditionally been explained by strategic changes of response criteria to less or more conservative thresholds. This idea has been implemented in current cognitive control frameworks, where it is proposed that high or low levels of processing conflict can dynamically change these response thresholds to achieve optimal performance. However, recent evidence suggests that evaluation of conflict is time consuming and can potentially interfere with subsequent processing [Jentzsch, I., Dudschig, C., 2009. Why do we slow down after an error? Mechanisms underlying the effects of posterror slowing. Quarterly Journal of Experimental Psychology, 62, 209-218]. The present study aims to extend this finding by investigating whether similar mechanisms underlie effects of pre-error speeding and posterror slowing and whether the amplitude of the Ne/ERN predicts posterror slowing in the current task setting. The response stimulus interval (RSI) was systematically manipulated. Speed-up in pre-error trials was unaffected by RSI, suggesting that this effect is not the result of strategic, time consuming control processes. Posterror slowing dramatically increased and performance became more error prone with decreasing RSI, providing further evidence for the idea that error evaluation can produce substantial interference with subsequent trial processing, particularly when there is insufficient time between the error and the subsequent event. Importantly, we did not find a positive relationship between the RSI-dependent change in posterror slowing and the Ne/ERN amplitude, questioning a direct link between the amplitude of this component and the amount of subsequent performance adjustments. PMID- 19679113 TI - Targeting ADAM10 to lipid rafts in neuroblastoma SH-SY5Y cells impairs amyloidogenic processing of the amyloid precursor protein. AB - ADAM10 (a disintegrin and metalloproteinase 10) is the principal alpha-secretase responsible for the non-amyloidogenic processing of the Alzheimer's disease (AD) associated amyloid precursor protein (APP). A reciprocal relationship exists between non-amyloidogenic and amyloidogenic APP processing such that impaired ADAM10-mediated proteolysis of the protein serves to enhance amyloidogenic processing (by beta- and gamma-secretases) thereby elevating levels of the amyloid beta (A beta)-peptides responsible for the neuronal death observed in the AD-afflicted brain. It has previously been demonstrated that the amyloidogenic processing of APP occurs within specialized regions of the cell membrane known as lipid rafts. Conversely, ADAM10-mediated non-amyloidogenic processing is thought to occur in the non-raft region of the membrane with the majority of ADAM10 being excluded from rafts. In the current study, we hypothesized that the exclusion of ADAM10 from rafts may leave the APP substrate particularly susceptible to alternative beta-secretase cleavage within these microdomains. In order to test this hypothesis, we targeted ADAM10 to rafts by replacing its transmembrane and cytosolic regions with a glycosylphosphatidylinositol (GPI) anchor and examined the associated effects on APP proteolysis. We found that whereas wild-type ADAM10 was exclusively present in the non-raft region of the membrane where it enhanced non-amyloidogenic APP proteolysis, GPI-anchored ADAM10 was effectively targeted to rafts where it competed with beta-secretase thereby reducing amyloidogenic APP processing. These results indicate that it is the exclusion of ADAM10 from rafts rather than simply the raft localization of beta- and gamma-secretases that underlies A beta-peptide generation within these cellular structures. PMID- 19679116 TI - Platelet-derived growth factor-BB reflects clinical, inflammatory and angiogenic disease activity and oxidative stress in inflammatory bowel disease. AB - OBJECTIVES: The goal of the studies was the evaluation of platelet-stored (serum) and circulating (plasma) pools of platelet-derived growth factor (PDGF)-BB in inflammatory bowel disease (IBD) and the assessment of a possible application of PDGF as the disease marker. DESIGN AND METHODS: Serum and plasma PDGF-BB were measured in 134 IBD patients and 81 controls and evaluated with respect to the disease status, endoscopic, inflammatory, and angiogenic activity. The diagnostic utility was evaluated using ROC analysis. RESULTS: PDGF was increased exclusively in active IBD regardless the disease type and associated with its clinical and endoscopic activity. Serum- and plasma-PDGF were poorly interrelated. Plasma-PDGF better reflected oxidative stress whereas serum-PDGF reflected inflammation and angiogenesis. In multivariate analysis, platelets alone explained about 30% in the PDGF variability and seemed to mediate most of the observed relationships. CONCLUSIONS: IBD is associated with the increases in platelet-stored and circulating PDGF, which correspond with the disease clinical, endoscopic, inflammatory, and angiogenic activity and IBD-associated oxidative stress. However, PDGF as an active-IBD marker was not better than currently applied C reactive protein, erythrocyte sedimentation rate and platelets. PMID- 19679115 TI - Crystalline silica Min-U-Sil 5 induces oxidative stress in human bronchial epithelial cells BEAS-2B by reducing the efficiency of antiglycation and antioxidant enzymatic defenses. AB - Reactive oxygen species (ROS) play an important role as mediators of pulmonary damage in mineral dust-induced diseases. Studies carried out to date have largely focused on silica-induced production of ROS by lung phagocytes. In this study we investigated the hypothesis that crystalline silica Min-U-Sil 5 can induce elevations in intracellular ROS in human bronchial epithelial cells BEAS-2B, via an indirect mechanism that involves ROS-inducing intracellular factors, through a reduction of antiglycation (glyoxalase enzymes) and antioxidant (paraoxonase 1 and glutathione-S-transferases) enzymatic defenses. The results show that crystalline silica Min-U-Sil 5 causes a significant reduction in the efficiency of antiglycation and antioxidant enzymatic defenses, paralleled by an early and extensive ROS generation, thus preventing the cells from an efficient scavenging action, and eliciting oxidative damage. These results confirm the importance of ROS in development of crystalline silica-induced oxidative stress and emphasize the pivotal role of antiglycation/antioxidant and detoxifying systems in determining the level of protection from free radicals-induced injury for cells exposed to crystalline silica Min-U-Sil 5. PMID- 19679117 TI - Oxidative stress and DNA damage in older adults that do exercises regularly. AB - OBJECTIVES: Free radicals may damage lipids, proteins and DNA, which may lead to critical diseases in the aging. This work evaluated levels of malondialdehyde (MDA), glutathione peroxidase (GPx) and DNA damage by comet assay (SCGE) in older adults that do exercises regularly. DESIGN AND METHODS: 110 females, aged 66.3+/ 8 years were divided into sedentary (n=54), walking (n=36) and muscle building (n=20) groups. Levels of MDA, GPx and SCGE were measured in venous blood before and after exercise. RESULTS: MDA levels were higher (P<0.005) and GPx levels were lower (P<0.005) in active groups than in sedentary group. SCGE index after physical activity was greater than at baseline (muscle building: P=0.004; walking: P=0.002). CONCLUSIONS: Exercise reduces the diseases risk, but may promote the production of free radicals. It remains unclear whether cell adaptations responsible for health benefits are associated with such events. However we may suggest the existence of a different biochemical pattern for older adults that do exercise regularly. PMID- 19679118 TI - The cis-regulatory system of the tbrain gene: Alternative use of multiple modules to promote skeletogenic expression in the sea urchin embryo. AB - The genomic cis-regulatory systems controlling regulatory gene expression usually include multiple modules. The regulatory output of such systems at any given time depends on which module is directing the function of the basal transcription apparatus, and ultimately on the transcription factor inputs into that module. Here we examine regulation of the Strongylocentrotus purpuratus tbrain gene, a required activator of the skeletogenic specification state in the lineage descendant from the embryo micromeres. Alternate cis-regulatory modules were found to convey skeletogenic expression in reporter constructs. To determine their relative developmental functions in context, we made use of recombineered BAC constructs containing a GFP reporter and of derivatives from which specific modules had been deleted. The outputs of the various constructs were observed spatially by GFP fluorescence and quantitatively over time by QPCR. In the context of the complete genomic locus, early skeletogenic expression is controlled by an intron enhancer plus a proximal region containing a HesC site as predicted from network analysis. From ingression onward, however, a dedicated distal module utilizing positive Ets1/2 inputs contributes to definitive expression in the skeletogenic mesenchyme. This module also mediates a newly discovered negative Erg input which excludes non-skeletogenic mesodermal expression. PMID- 19679119 TI - Protein kinase VRK-1 regulates cell invasion and EGL-17/FGF signaling in Caenorhabditis elegans. AB - The vaccinia-related kinases (VRKs) are highly conserved throughout the animal kingdom and phosphorylate several chromatin proteins and transcription factors. In early Caenorhabditis elegans embryos, VRK-1 is required for proper nuclear envelope formation. In this work, we present the first investigation of the developmental role of VRKs by means of a novel C. elegans vrk-1 mutant allele. We found that VRK-1 is essential in hermaphrodites for formation of the vulva, uterus, and utse and for development and maintenance of the somatic gonad and thus the germ line. VRK-1 regulates anchor cell polarity and the timing of anchor cell invasion through the basement membranes separating vulval and somatic gonadal cells during the L3 larval stage. VRK-1 is also required for proper specification and proliferation of uterine cells and sex myoblasts. Expression of the fibroblast growth factor-like protein EGL-17 and its receptor EGL-15 is reduced in vrk-1 mutants, suggesting that VRK-1 might act at least partially through activation of FGF signaling. Expression of a translational VRK-1Colon, two colonsGFP fusion protein in the ventral nerve cord and vulva precursor cells restores vulva and uterus formation, suggesting both cell autonomous and non autonomous roles of VRK-1. PMID- 19679122 TI - Classic Kaposi sarcoma with sarcoid-like granulomas: a case report and literature review. AB - The occurrence of granulomatous reactions within lymph nodes that drain carcinomas is well known but uncommon. Even more rarely, granulomas may occur within the stroma of tumors. These lesions, called sarcoid-like reactions, commonly accompany carcinomas but are very rare in sarcomas. This study presents a case of classic Kaposi sarcoma that contained sarcoid-like granulomas, with a literature review. A soft tissue lesion of the foot was excised from a 74-year old male. Histopathological examination showed that the tumor tissue consisted of spindle cells, areas of atypical vascular proliferation, and extravasated erythrocytes surrounded by non-caseating granulomas. The patient had no clinical or laboratory findings of sarcoidosis. The case was interpreted as "Kaposi sarcoma containing sarcoid-like granulomas". The association of soft tissue sarcomas with a granulomatous reaction is very rare. A granulomatous reaction is reported to be a good prognostic indicator in several carcinoma types, although its importance in sarcomas is unclear. PMID- 19679120 TI - Arabidopsis actin-related protein ARP5 in multicellular development and DNA repair. AB - Actin-related protein 5 (ARP5) is a conserved subunit of the INO80 chromatin remodeling complex in yeast and mammals. We have characterized the expression and subcellular distribution of Arabidopsis thaliana ARP5 and explored its role in the epigenetic control of multicellular development and DNA repair. ARP5-specific monoclonal antibodies localized ARP5 protein to the nucleoplasm of interphase cells in Arabidopsis and Nicotiana tabacum. ARP5 promoter-reporter fusions and the ARP5 protein are ubiquitously expressed. A null mutant and a severe knockdown allele produced moderately dwarfed plants with all organs smaller than the wild type. The small and slightly deformed organs such as leaves and hypocotyls were composed of small-sized cells. The ratio of leaf stomata to epidermal cells was high in the mutant, which also exhibited a delayed stomatal development compared with the wild type. Mutant plants were hypersensitive to DNA-damaging reagents including hydroxyurea, methylmethane sulfonate, and bleocin, demonstrating a role for ARP5 in DNA repair. Interestingly, the hypersensitivity phenotype of ARP5 null allele arp5-1 is stronger than the severe knockdown allele arp5-2. Moreover, a wild-type transgene fully complemented all developmental and DNA repair mutant phenotypes. Despite the common participation of both ARP4 and ARP5 in the INO80 complex, ARP4- and ARP5-deficient plants displayed only a small subset of common phenotypes and each displayed novel phenotypes, suggesting that in Arabidopsis they have both shared and unique functions. PMID- 19679123 TI - Relationships between increased aqueous outflow facility during washout with the changes in hydrodynamic pattern and morphology in bovine aqueous outflow pathways. AB - Previous studies suggest that the structural correlate for the increased outflow facility (C) during washout in the bovine eye is separation between the inner wall (IW) and underlying juxtacanalicular connective tissue (JCT). However, how these structural changes affect hydrodynamic patterns of outflow during washout has not been studied. We hypothesize that an increase in the outflow facility during washout is associated with an increase in the effective filtration area (EFA) of aqueous outflow, which is regulated by a loss of the connectivity between the IW and JCT. To test this hypothesis, the relationship between C and the hydrodynamic patterns of outflow as well as the morphological changes in JCT and IW during the washout were investigated. Ten bovine eyes were perfused at 15 mmHg with Dulbecco's PBS + 5.5 mM glucose (DPBS) for 30 min to establish stable baseline C. After measuring baseline C, five eyes (short-duration group) were perfused with 0.5 mL DPBS containing 0.002% microspheres (0.5 microm) to trace the hydrodynamic pattern of outflow. Five other eyes (long-duration group) were perfused for 3 h to elicit a significant washout effect followed by subsequent perfusion of the same volume (0.5 mL) of microspheres to map out the outflow pattern after washout. All eyes were then perfusion-fixed. Anterior segments were sectioned and prepared for confocal and light microscopy. Total length (TL) and filtration length (FL) of the IW were measured in > or =15 images/eye to calculate percent effective filtration length (PEFL = FL/TL) while TL and length exhibiting JCT/IW separation (SL) were measured in > or =13 images/eye to calculate percent separation length (PSL = SL/TL). In long-duration eyes, C increased 170.5 +/- 21.3% (mean +/- SEM, 1.55 +/- 0.24 vs 4.13 +/- 0.55 microl/min/mmHg, p = 0.001) above baseline. Pre-fixation C (4.13 +/- 0.55 microl/min/mmHg) in long-duration was 1.6-fold greater than that (2.14 +/- 0.61 microl/min/mmHg; p = 0.042) in short-duration. A more uniform tracer labeling was observed in the JCT/IW of long-duration eyes compared to short-duration. PEFL was 2.3-fold larger (52.82 +/- 6.06 vs. 22.2 +/- 6.0%; p = 0.007) and PSL was 2.6 fold larger (54.2 +/- 6.0 vs. 20.5 +/- 1.3%; p = 0.004) in long-duration eyes compared to short-duration. Data from all eyes revealed a positive correlation between PEFL and PSL (p = 0.02). Both PEFL and PSL demonstrated significant positive correlations with the relative increase in C due to washout (p < or = 0.05). An additional experiment was performed in which unequal volumes of tracer (0.5 and 1.0 mL) were perfused in paired eyes for both short- and long-duration (N = 2 for each condition) to examine the affect on PEFL. No significant change in PEFL was found in eyes perfused with 0.5 and 1.0 mL within the same group. These data support our hypothesis that separations between the IW and JCT result in an increase in the EFA that then influences C. Altogether, these data suggest that outflow hydrodynamics and the tissue structure work together to regulate outflow resistance. PMID- 19679121 TI - Stripy Ftz target genes are coordinately regulated by Ftz-F1. AB - During development, cascades of regulatory genes act in a hierarchical fashion to subdivide the embryo into increasingly specified body regions. This has been best characterized in Drosophila, where genes encoding regulatory transcription factors form a network to direct the development of the basic segmented body plan. The pair-rule genes are pivotal in this process as they are responsible for the first subdivision of the embryo into repeated metameric units. The Drosophila pair-rule gene fushi tarazu (ftz) is a derived Hox gene expressed in and required for the development of alternate parasegments. Previous studies suggested that Ftz achieves its distinct regulatory specificity as a segmentation protein by interacting with a ubiquitously expressed cofactor, the nuclear receptor Ftz-F1. However, the downstream target genes regulated by Ftz and other pair-rule genes to direct segment formation are not known. In this study, we selected candidate Ftz targets by virtue of their early expression in Ftz-like stripes. This identified two new Ftz target genes, drumstick (drm) and no ocelli (noc), and confirmed that Ftz regulates a serotonin receptor (5-HT2). These are the earliest Ftz targets identified to date and all are coordinately regulated by Ftz-F1. Engrailed (En), the best-characterized Ftz/Ftz-F1 downstream target, is not an intermediate in regulation. The drm genomic region harbors two separate seven stripe enhancers, identified by virtue of predicted Ftz-F1 binding sites, and these sites are necessary for stripe expression in vivo. We propose that pair rule genes, exemplified by Ftz/Ftz-F1, promote segmentation by acting at different hierarchical levels, regulating first, other segmentation genes; second, other regulatory genes that in turn control specific cellular processes such as tissue differentiation; and, third, 'segmentation realizator genes' that are directly involved in morphogenesis. PMID- 19679124 TI - A peek inside the Huntington's brain: will functional imaging take us one step closer in solving the puzzle? PMID- 19679125 TI - Pathological role of hypoxia in Alzheimer's disease. AB - The majority cases of Alzheimer's disease (AD) are sporadic late-onset form not being linked to APP and PS1 gene mutations. It is believed that the environmental risk factors play an important role in the onset and development of AD. Patients suffering from cerebral ischemia and stroke in which hypoxic conditions occur are much more susceptible to AD. Increasing evidence suggests that hypoxia facilitates the pathogenesis of AD through accelerating the accumulation of Abeta, increasing the hyperphosphorylation of tau, impairing the normal functions of blood-brain barrier, and promoting the degeneration of neurons. Further investigations into the relationship between hypoxia and AD may open the avenue for effective preservation and pharmacological treatments of this neurodegenerative disease. PMID- 19679126 TI - Anti-inflammatory treatment in AD mice protects against neuronal pathology. AB - Prior studies suggest that non-steroidal anti-inflammatory drugs (NSAIDs) may lower the incidence of Alzheimer's disease (AD) and delay onset or slow progression of symptoms in mouse models of AD. We examined the effects of chronic NSAID treatment in order to determine which elements of the pathological features might be ameliorated. We compared the effects of the NSAIDs ibuprofen and celecoxib on immunohistological and neurochemical markers at two different ages in APPxPS1 mice using measurements of amyloid plaque deposition, Abeta peptide levels, and neurochemical profiles using magnetic resonance spectroscopy (MRS). At 6 months of age, few neurochemical changes were observed between PSAPP mice and WT mice using MRS. Ibuprofen, but not celecoxib, treatment significantly decreased the Abeta(42/40) ratio in frontal cortex at 6 months, but overall amyloid plaque burden was unchanged. Consistent with prior findings in mouse models, at 17 months of age, there was a decrease in the neuronal markers NAA and glutamate and an increase in the astrocytic markers glutamine and myo-inositol in AD mice compared to WT. Ibuprofen provided significant protection against NAA and glutamate loss. Neither of the drugs significantly affected myo-inositol or glutamine levels. Both ibuprofen and celecoxib lowered plaque burden without a significant effect on Abeta(1-42) levels. NAA levels significantly correlated with plaque burden. These results suggest that selective NSAIDs (ibuprofen and possibly celecoxib) treatment can protect against the neuronal pathology. PMID- 19679128 TI - Inhibitory effect of essential oils of Allium sativum and Piper longum on spontaneous muscular activity of liver fluke, Fasciola gigantica. AB - Effects of essential oil of Allium sativum (garlic) and Piper longum (Indian long pepper) were evaluated on muscular activity of whole Fasciola gigantica and its strip preparation. The whole flukes and longitudinal strip preparations of the flukes were isometrically mounted to record the spontaneous muscular activity (SMA) and to evaluate effects of cumulative doses (0.1, 0.3, 1.0 and 3.0mg/ml) of the plant essential oils. Whole flukes and the strip preparations exhibited continuous SMA without any significant difference in its baseline tension, frequency and amplitude for 2h. Essential oil of A. sativum produced significant reduction in the frequency and the amplitude of the SMA of whole fluke at 1 and 3mg/ml concentrations. It caused complete paralysis of the fluke after 15 min of administration of 3mg/ml concentration. Similar to whole fluke, essential oil of A. sativum (3mg/ml) also produced flaccid paralysis in the strip preparations of the flukes. Essential oil of P. longum firstly induced marked excitatory effect and then there was flaccid paralysis of the whole fluke following 15 min exposure at 3mg/ml concentration. Complete flaccid paralysis of the strip preparation was also ensued after 15 min of administration of 3mg/ml concentration of P. longum. In both the essential oils, the whole fluke and strip preparations did not recover from paralysis following 2-3 washes. In conclusion, the observations demonstrated irreversible paralytic effect of essential oils of A. sativum and P. longum on F. giganticain vitro which might possibly help to developing herbal based anthelmintic. PMID- 19679127 TI - Volumetric effects of motor cortex injury on recovery of dexterous movements. AB - Due to the heterogeneous nature of most brain injuries, the contributions of gray and white matter involvement to motor deficits and recovery potential remain obscure. We tested the hypothesis that duration of hand motor impairment and recovery of skilled arm and hand motor function depends on the volume of gray and white matter damage of the frontal lobe. Lesions of the primary motor cortex (M1), M1 + lateral premotor cortex (LPMC), M1 + LPMC + supplementary motor cortex (M2) or multifocal lesions affecting motor areas and medial prefrontal cortex were evaluated in rhesus monkeys. Fine hand motor function was quantitatively assessed pre-lesion and for 3-12 months post-lesion using two motor tests. White and gray matter lesion volumes were determined using histological and quantitative methods. Regression analyses showed that duration of fine hand motor impairment was strongly correlated (R(2)>0.8) with the volume of gray and white matter lesions, with white matter lesion volume being the primary predictor of impairment duration. Level of recovery of fine hand motor skill was also well correlated (R(2)>0.5) with gray and white matter lesion volume. In some monkeys post-lesion skill exceeded pre-lesion skill in one or both motor tasks demonstrating that continued post-injury task practice can improve motor performance after localized loss of frontal motor cortex. These findings will assist in interpreting acute motor deficits, predicting the time course and expected level of functional recovery, and designing therapeutic strategies in patients with localized frontal lobe injury or neurosurgical resection. PMID- 19679129 TI - Determination of LC(50) of permethrin acaricide in semi-engorged females of the tick Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae). AB - Currently, the most efficient and widely used method of tick control still is the treatment with acaricides, especially permethrin (active ingredient of the Advantage, Max3, Bayer), a pyrethroid with neurotoxic action. Due to the wide use of this acaricide in the control of the tick Rhipicephalus sanguineus, this study carried out laboratorial procedures to determine the LC(50) (lethal concentration fifty) of permethrin in semi-engorged females of R. sanguineus. Based on the result of 14 dilutions of permethrin in distilled water and later Probit analysis, the LC(50) of permethrin for R. sanguineus was 2062 ppm (1549-2675 ppm). This work can be used as a protocol with other chemicals, to determinate the LC(50), basic procedure for studies of control, resistance and behavior of ticks treated with acaricides, especially the brown dog tick R. sanguineus. Also, the knowledge of the LC(50) provides information on the potency of chemicals, the sensitivity of Arthropods to them and even estimates on pest control. PMID- 19679130 TI - Proteomic analysis of wild type and arsenite-resistant Leishmania donovani. AB - Leishmania donovani, causative organism for visceral leishmaniasis, is responsible for considerable mortality and morbidity worldwide. Generation of drug-resistant variants continue to challenge the chemotherapy, the mainstay to fight the disease. The aim of current study was proteomic profiling of wild type (Ld-Wt) and arsenite-resistant (Ld-As20) L. donovani. Significant differences in protein profiles were observed between Ld-As20 and its parent Ld-Wt strain. Proteomic analysis of 158 spots from Ld-Wt and 144 spots from, Ld-As20 identified 77 and 74 protein entries, respectively, through MALDI-TOF/TOF based mass spectrometry and database search. A shift in the isoelectric point of few proteins was observed both in Ld-Wt and Ld-As20, which raises the possibility of continuous arsenite stress, resulting in the differences in the protein profiles of drug-resistant strain from its parent wild type strain. The comparative proteomic data holds the key for elucidation of the multifactorial and complex drug resistance mechanism, like arsenite resistance, in the parasite. PMID- 19679131 TI - Similar distribution of simple sequence repeats in diverse completed Human Immunodeficiency Virus Type 1 genomes. AB - The survey of simple sequence repeats (SSRs) has been extensively made in eukaryotes and prokaryotes. However, its still rare in viruses. Thus, we undertook a survey of SSRs in Human Immunodeficiency Virus Type 1 (HIV-1) which is an excellent system to study evolution and roles of SSRs in viruses. Distribution of SSRs was examined in 81 completed HIV-1 genome sequences which come from 34 different countries or districts over 6 continents. In these surveyed sequences, although relative abundance and relative density exhibit very high similarity, some of these sequences show different preference for most common SSRs and longest SSRs. Our results suggest proportion of various repeat types might be related to genome stability. PMID- 19679132 TI - Male sexual behavior contributes to the maintenance of high LH pulsatility in anestrous female goats. AB - The objective of this study was to determine the importance of male sexual behavior in stimulating LH secretion in anovulatory female goats. Two groups of females (n=10 per group) were each exposed to a buck in sexual rest and submitted to natural daylength. In one group, the buck was awake, whereas in the other group, it was sedated to prevent its sexual behavior. Two other groups of goats (n=10 per group) were exposed to sexually active bucks that had been exposed to 2.5 months of long days. In one group, the buck was awake, and in the other group, it was sedated. LH secretion was determined every 15 min from 4 h before introducing the bucks to 8 h after, then every 15 min again from 20 to 24 h after introducing the bucks. The bucks submitted to natural daylength did not stimulate LH secretion (P>0.05), whether they were sedated or not. In contrast, both the awake and the sedated light-treated bucks induced an increase (P<0.05) of LH pulsatility in the first 4 h following their introduction. However, pulsatility remained elevated until 24 h in the females exposed to the light-treated awake buck, whereas in the group with the light-treated sedated buck, pulsatility diminished (P<0.05) after the first 4 h of stimulation by the buck. In conclusion, the sexual behavior of males contributes to the maintenance of a high LH pulsatility up to 24 h after introduction into a group of anovulatory goats. PMID- 19679133 TI - Acid phosphatase activity during the interaction of the nematophagous fungus Duddingtonia flagrans with the nematode Panagrellus sp. AB - The use of Duddingtonia flagrans, a nematode-trapping fungus, has been investigated as a biological control method against free living larvae of gastrointestinal nematodes of livestock animals. This fungus captures and infects the nematode by cuticle penetration, immobilization and digestion of the internal contents. It has been suggested that this sequence of events occurs by a combination of physical and enzymatical activities. This report characterizes the acid phosphatase activity during the interaction of D. flagrans with the free living nematode Panagrellus sp. The optimum pH for the hydrolysis of the acid phosphatase substrate p-nitrophenyl phosphate was 2.2, 2.8 and 5.4 from D. flagrans alone and 2.2 and 5.4 for Panagrellus sp alone, fungus-nematode interaction in liquid medium and fungus-nematode interaction in solid medium. Different acid phosphatase activity bands were detected by SDS-PAGE. Maximum acid phosphatase activity of the fungus or nematode alone and of the fungus-nematode interaction occurred within 70min of incubation in the presence of the substrate 4-methylumbelliferyl phosphate. The activity of this enzyme was significantly higher for the fungus-nematode interaction when compared to the organisms alone, indicating a synergistic response. Furthermore, structures appeared in the hyphae after 30min, nematodes were observed adhered after 40min and many were captured by the typical fungus traps after 70min of interaction. The participation of acid phosphatase activity and its importance during the interaction of the fungus with the nematode were discussed. PMID- 19679134 TI - Ecological characterization of entomopathogenic nematodes isolated in stone fruit orchard soils of Mediterranean areas. AB - Entomopathogenic nematodes in the families Steinernematidae and Heterorhabditidae were isolated from stone-fruit orchards in two Mediterranean regions of Spain. A total of 630 soil samples (210 sites) from Catalonia and 90 soil samples (30 sites) from Murcia were evaluated resulting in 5.2% and 20% of the soils testing positive for nematodes, respectively. Ten steinernematid isolates and three heterorhabditid isolates were recovered using the Galleria mellonella baiting method. Based on morphometric data, molecular data, and cross-breeding experiments the nematode species were identified as Steinernemafeltiae and Heterorhabditis bacteriophora. Environmental tolerance to heat, desiccation and hypoxia, the effect of temperature on infectivity and reproduction and nematode migration in sand columns were compared among isolates and one Steinernema carpocapsae strain. Results showed differences among species and a great variability within species. Beneficial traits for each strain were added up to identify a superior candidate to control Mediterranean flat-headed rootborer, Capnodis tenebrionis. When all analyzed factors were considered, three S. feltiae isolates (Bpa, Sor and M116) obtained the best scores, and when hypoxia was removed, two of the strains (Bpa and Sor) continued ranking superior to other strains. PMID- 19679135 TI - Activation of purinergic receptors by ATP induces ventricular tachycardia by membrane depolarization and modifications of Ca2+ homeostasis. AB - Cardiac myocytes are continuously exposed to extracellular nucleotides secreted by the myocytes themselves, nerve terminals, or platelets and other blood cells during coronary perfusion, and the concentrations of such extracellular nucleotides are known to increase during cardiac ischemia and hypoxia. The effects of the extracellular nucleotides ATP, ADP, UTP, and adenosine on ventricular arrhythmogenic properties were explored in 36 Langendorff-perfused mouse hearts using monophasic action potential recording. Extracellular nucleotides induced arrhythmic phenomena in form of ectopic activity and ventricular tachycardia in a potency order of ATP (n=7) > ADP (n=5) > UTP (n=3) approximately adenosine (n=3). The purinergic receptor antagonists suramin and pyridoxal phosphate-6-azo(benzene-2,4-disulphonic acid) reduced the incidence of ATP-triggered arrhythmias. In isolated ventricular myocytes, ATP induced sustained increases in diastolic Ca2+ and triggered multiple Ca2+ waves, which were inhibited by suramin but not by the L-type Ca2+ channel antagonist nifedipine. In whole-cell patch clamp experiments, extracellular ATP induced two distinct types of inward currents, which were inhibited by suramin and PPADS, suggesting activation of P2X receptors. ATP also induced delayed after depolarizations and ectopic action potentials in current clamped ventricular myocytes. In conclusion, extracellular ATP activates purinergic receptors and induces arrhythmic activity through modifications of Ca2+ homeostasis and an activation of depolarizing membrane currents. PMID- 19679137 TI - TN curve: a novel 3D graphical representation of DNA sequence based on trinucleotides and its applications. AB - In this paper, a novel 3D graphical representation of DNA sequence based on trinucleotides is proposed. This representation allows direct inspection of composition as well as distribution of trinucleotides in DNA sequence for the first time and avoids loss of information, from which one can obtain more information. Based on this novel model, six numerical descriptors of DNA sequence are deduced without complicated calculations, and the applications in similarities/dissimilarities analysis of coding sequences and conserved genes discrimination illustrate their utilities. In addition, two simple methods for similarities/dissimilarities analysis of coding sequences among different species are exploited by using two vectors composed of 64 and six components, respectively, which can provide convenient sequence alignment tools for both computational scientists and molecular biologists. PMID- 19679136 TI - A model of interactions between radiation-induced oxidative stress, protein and DNA damage in Deinococcus radiodurans. AB - Ionizing radiation triggers oxidative stress, which can have a variety of subtle and profound biological effects. Here we focus on mathematical modeling of potential synergistic interactions between radiation damage to DNA and oxidative stress-induced damage to proteins involved in DNA repair/replication. When sensitive sites on these proteins are attacked by radiation-induced radicals, correct repair of dangerous DNA lesions such as double strand breaks (DSBs) can be compromised. In contrast, if oxidation of important proteins is prevented by strong antioxidant defenses, DNA repair may function more efficiently. These processes probably occur to some extent even at low doses of radiation/oxidative stress, but they are easiest to investigate at high doses, where both DNA and protein damage are extensive. As an example, we use data on survival of Deinococcus radiodurans after high doses (thousands of Gy) of acute and chronic irradiation. Our model of radiogenic oxidative stress is consistent with these data and can potentially be generalized to other organisms and lower radiation doses. PMID- 19679138 TI - SubChlo: predicting protein subchloroplast locations with pseudo-amino acid composition and the evidence-theoretic K-nearest neighbor (ET-KNN) algorithm. AB - The chloroplast is a type of plant specific subcellular organelle. It is of central importance in several biological processes like photosynthesis and amino acid biosynthesis. Thus, understanding the function of chloroplast proteins is of significant value. Since the function of chloroplast proteins correlates with their subchloroplast locations, the knowledge of their subchloroplast locations can be very helpful in understanding their role in the biological processes. In the current paper, by introducing the evidence-theoretic K-nearest neighbor (ET KNN) algorithm, we developed a method for predicting the protein subchloroplast locations. This is the first algorithm for predicting the protein subchloroplast locations. We have implemented our algorithm as an online service, SubChlo (http://bioinfo.au.tsinghua.edu.cn/subchlo). This service may be useful to the chloroplast proteome research. PMID- 19679139 TI - Inbreeding, pedigree size, and the most recent common ancestor of humanity. AB - How many generations ago did the common ancestor of all present-day individuals live, and how does inbreeding affect this estimate? The number of ancestors within family trees determines the timing of the most recent common ancestor of humanity. However, mating is often non-random and inbreeding is ubiquitous in natural populations. Rates of pedigree growth are found for multiple types of inbreeding. This data is then combined with models of global population structure to estimate biparental coalescence times. When pedigrees for regular systems of mating are constructed, the growth rates of inbred populations contain Fibonacci n-step constants. The timing of the most recent common ancestor depends on global population structure, the mean rate of pedigree growth, mean fitness, and current population size. Inbreeding reduces the number of ancestors in a pedigree, pushing back global common ancestry times. These results are consistent with the remarkable findings of previous studies: all humanity shares common ancestry in the recent past. PMID- 19679140 TI - Membrane-spanning peptides and the origin of life. AB - An explanation is given as to why membrane-spanning peptides must have been the first "information-rich" molecules in the development of life. These peptides are stabilised in a lipid bilayer membrane environment and they are preferentially made from the simplest, and likewise oldest, of the amino acids that survive today. Transmembrane peptides can exercise functions that are essential for biological systems such as signal transduction and material transport across membranes. More complex peptides possessing catalytic properties could later develop on either side of the membrane as independently folding functional units formed by extension of the protruding ends of the transmembrane peptides within an aqueous environment and thereby give rise to more of the functions that are necessary for life. But the membrane was the cradle for the development of the first information-rich biomolecules. PMID- 19679141 TI - Epidemics with general generation interval distributions. AB - We study the spread of susceptible-infected-recovered (SIR) infectious diseases where an individual's infectiousness and probability of recovery depend on his/her "age" of infection. We focus first on early outbreak stages when stochastic effects dominate and show that epidemics tend to happen faster than deterministic calculations predict. If an outbreak is sufficiently large, stochastic effects are negligible and we modify the standard ordinary differential equation (ODE) model to accommodate age-of-infection effects. We avoid the use of partial differential equations which typically appear in related models. We introduce a "memoryless" ODE system which approximates the true solutions. Finally, we analyze the transition from the stochastic to the deterministic phase. PMID- 19679142 TI - Plant competition and exclusion with optimizing individuals. AB - Most models of plant competition represent competition as taking place between species when realistically competition takes place between individuals. We model individual plants as optimally choosing biomass in order to maximize net energy that is directed into reproduction. Competition is for access to light and a plant that grows more biomass adds to the leaf area index, creating negative feedback in the form of more self shading and shading of its neighbors. In each period and for given species densities, simultaneous maximization by all plants yields an equilibrium characterized by optimum biomasses. Between periods the net energies plants obtain are used to update the densities, and if densities change the equilibrium changes in the subsequent period. A steady state is attained when all plants have net energies that just allow for replacement. Four main predictions of the resource-ratio theory of competition are obtained, providing behavioral underpinnings for species level models. However, if individual plant parameters are not identical across species, then the predictions do not follow. The optimization framework yields many other predictions, including how specific leaf areas and resource stress impact biomass and leaf area indices. PMID- 19679143 TI - Evolution under the multilocus Levene model without epistasis. AB - Evolution under the multilocus Levene model is investigated. The linkage map is arbitrary, but epistasis is absent. The geometric-mean fitness, w(rho), depends only on the vector of gene frequencies, rho; it is nondecreasing, and the single generation change is zero only on the set, Lambda, of gametic frequencies at gene frequency equilibrium. The internal gene-frequency equilibria are the stationary points of w(rho). If the equilibrium points rho of rho(t) (where t denotes time in generations) are isolated, as is generic, then rho(t) converges as t- >infinity to some rho. Generically, rho(t) converges to a local maximum of w(rho). Write the vector of gametic frequencies, p, as (rho,d)(T), where d represents the vector of linkage disequilibria. If rho is a local maximum of w(rho), then the equilibrium point (rho,0)(T) is asymptotically stable. If either there are only two loci or there is no dominance, then d(t)-->0 globally as t- >infinity. In the second case, w(rho) has a unique maximum rho and (rho,0)(T) is globally asymptotically stable. If underdominance and overdominance are excluded, and if at each locus, the degree of dominance is deme independent for every pair of alleles, then the following results also hold. There exists exactly one stable gene-frequency equilibrium (point or manifold), and it is globally attracting. If an internal gene-frequency equilibrium exists, it is globally asymptotically stable. On Lambda, (i) the number of demes, Gamma, is a generic upper bound on the number of alleles present per locus; and (ii) if every locus is diallelic, generically at most Gamma-1 loci can segregate. Finally, if migration and selection are completely arbitrary except that the latter is uniform (i.e., deme independent), then every uniform selection equilibrium is a migration-selection equilibrium and generically has the same stability as under pure selection. PMID- 19679144 TI - Differential pulmonary and cardiac effects of pulmonary exposure to a panel of particulate matter-associated metals. AB - Biological mechanisms underlying the association between particulate matter (PM) exposure and increased cardiovascular health effects are under investigation. Water-soluble metals reaching systemic circulation following pulmonary exposure are likely exerting a direct effect. However, it is unclear whether specific PM associated metals may be driving this. We hypothesized that exposure to equimolar amounts of five individual PM-associated metals would cause differential pulmonary and cardiac effects. We exposed male WKY rats (14 weeks old) via a single intratracheal instillation (IT) to saline or 1 micromol/kg body weight of zinc, nickel, vanadium, copper, or iron in sulfate form. Responses were analyzed 4, 24, 48, or 96 h after exposure. Pulmonary effects were assessed by bronchoalveolar lavage fluid levels of total cells, macrophages, neutrophils, protein, albumin, and activities of lactate dehydrogenase, gamma-glutamyl transferase, and n-acetyl glucosaminidase. Copper induced earlier pulmonary injury/inflammation, while zinc and nickel produced later effects. Vanadium or iron exposure induced minimal pulmonary injury/inflammation. Zinc, nickel, or copper increased serum cholesterol, red blood cells, and white blood cells at different time points. IT of nickel and copper increased expression of metallothionein-1 (MT-1) in the lung. Zinc, nickel, vanadium, and iron increased hepatic MT-1 expression. No significant changes in zinc transporter-1 (ZnT-1) expression were noted in the lung or liver; however, zinc increased cardiac ZnT-1 at 24 h, indicating a possible zinc-specific cardiac effect. Nickel exposure induced an increase in cardiac ferritin 96 h after IT. This data set demonstrating metal-specific cardiotoxicity is important in linking metal enriched anthropogenic PM sources with adverse health effects. PMID- 19679145 TI - Effects of phenol on barrier function of a human intestinal epithelial cell line correlate with altered tight junction protein localization. AB - Phenol contamination of soil and water has raised concerns among people living near phenol-producing factories and hazardous waste sites containing the chemical. Phenol, particularly in high concentrations, is an irritating and corrosive substance, making mucosal membranes targets of toxicity in humans. However, few data on the effects of phenol after oral exposure exist. We used an in vitro model employing human intestinal epithelial cells (SK-CO15) cultured on permeable supports to examine effects of phenol on epithelial barrier function. We hypothesized that phenol disrupts epithelial barrier by altering tight junction (TJ) protein expression. The dose-response effect of phenol on epithelial barrier function was determined using transepithelial electrical resistance (TER) and FITC-dextran permeability measurements. We studied phenol induced changes in cell morphology and expression of several tight junction proteins by immunofluorescence and Western blot analysis. Effects on cell viability were assessed by MTT, Trypan blue, propidium iodide and TUNEL staining. Exposure to phenol resulted in decreased TER and increased paracellular flux of FITC-dextran in a dose-dependent manner. Delocalization of claudin-1 and ZO-1 from TJs to cytosol correlated with the observed increase in permeability after phenol treatment. Additionally, the decrease in TER correlated with changes in the distribution of a membrane raft marker, suggesting phenol-mediated effects on membrane fluidity. Such observations were independent of effects of phenol on cell viability as enhanced permeability occurred at doses of phenol that did not cause cell death. Overall, these findings suggest that phenol may affect transiently the lipid bilayer of the cell membrane, thus destabilizing TJ containing microdomains. PMID- 19679147 TI - Cascaded Bayesian processes: an account of bias in orientation perception. AB - Following adaptation to an oriented (1-d) signal in central vision, the orientation of subsequently viewed test signals may appear repelled away from or attracted towards the adapting orientation. Small angular differences between the adaptor and test yield 'repulsive' shifts, while large angular differences yield 'attractive' shifts. In peripheral vision, however, both small and large angular differences yield repulsive shifts. To account for these tilt after-effects (TAEs), a cascaded model of orientation estimation that is optimized using hierarchical Bayesian methods is proposed. The model accounts for orientation bias through adaptation-induced losses in information that arise because of signal uncertainties and neural constraints placed upon the propagation of visual information. Repulsive (direct) TAEs arise at early stages of visual processing from adaptation of orientation-selective units with peak sensitivity at the orientation of the adaptor (theta). Attractive (indirect) TAEs result from adaptation of second-stage units with peak sensitivity at theta and theta+90 degrees , which arise from an efficient stage of linear compression that pools across the responses of the first-stage orientation-selective units. A spatial orientation vector is estimated from the transformed oriented unit responses. The change from attractive to repulsive TAEs in peripheral vision can be explained by the differing harmonic biases resulting from constraints on signal power (in central vision) versus signal uncertainties in orientation (in peripheral vision). The proposed model is consistent with recent work by computational neuroscientists in supposing that visual bias reflects the adjustment of a rational system in the light of uncertain signals and system constraints. PMID- 19679146 TI - Spatio-temporal tuning of coherent motion evoked responses in 4-6 month old infants and adults. AB - Motion cues provide a rich source of information about translations of the observer through the environment as well as the movements of objects and surfaces. While the direction of motion can be extracted locally these local measurements are, in general, insufficient for determining object and surface motions. To study the development of local and global motion processing mechanisms, we recorded Visual Evoked Potentials (VEPs) in response to dynamic random dot displays that alternated between coherent rotational motion and random motion at 0.8 Hz. We compared the spatio-temporal tuning of the evoked response in 4-6 months old infants to that of adults by recording over a range of dot displacements and temporal update rates. Responses recorded at the frequency of the coherent motion modulation were tuned for displacement at the occipital midline in both adults in infants. Responses at lateral electrodes were tuned for speed in adults, but not in infants. Infant responses were maximal at a larger range of spatial displacement than that of adults. In contrast, responses recorded at the dot-update rate showed a more similar parametric displacement tuning and scalp topography in infants and adults. Taken together, our results suggest that while local motion processing is relatively mature at 4-6 months, global integration mechanisms exhibit significant immaturities at this age. PMID- 19679148 TI - Genetic and environmental contributions to strabismus and phoria: evidence from twins. AB - The causes of manifest (strabismus) and latent (phoria) misalignment of the visual axes are incompletely understood. We calculated genetic and environmental contributions to strabismus based upon a critical review and quantitative meta analysis of previous strabismus twin studies (n=3418 twin pairs) and calculated contributions to phoria based upon a new twin study (n=307 twin pairs). Our results suggest that genetic liability is necessary to develop strabismus, whereas environmental factors are sufficient to cause most phorias. The different etiologies implied by this work suggest that strabismus and phoria should be carefully distinguished in epidemiological work. PMID- 19679150 TI - Real-time PCR and enrichment culture for sensitive detection and enumeration of Escherichia coli. AB - Rapid enumeration of Escherichia coli strains by quantitative real-time PCR targeting the uidA gene was developed and confirmed for minced beef, tuna and raw oyster. Higher sensitivity (1 CFU/g of E. coli in all three food samples) was obtained by incubating for 7 h in TSB. Colony-directed E. coli specific TaqMan PCR assay could effectively distinguish colonies grown on various selective media within 1.5-h. Inspection of E. coli in food testing laboratories is important, and our rapid E. coli detection strategy will contribute to quality control in food industries. PMID- 19679149 TI - Transcriptional profiling of the age-related response to genotoxic stress points to differential DNA damage response with age. AB - The p53 DNA damage response attenuated with age and we have evaluated downstream factors in the DNA damage response. In old animals p21 protein accumulates in the whole cell fraction but significantly declines in the nucleus, which may alter cell cycle and apoptotic programs in response to DNA damage. We evaluated the transcriptional response to DNA damage in young and old and find 2692 genes are differentially regulated in old compared to young in response to oxidative stress (p<0.005). As anticipated, the transcriptional profile of young mice is consistent with DNA damage induced cell cycle arrest while the profile of old mice is consistent with cell cycle progression in the presence of DNA damage, suggesting the potential for catastrophic accumulation of DNA damage at the replication fork. Unique sets of DNA repair genes are induced in response to damage in old and young, suggesting the types of damage accumulating differs between young and old. The DNA repair genes upregulated in old animals point to accumulation of replication-dependent DNA double strand breaks (DSB). Expression data is consistent with loss of apoptosis following DNA damage in old animals. These data suggest DNA damage responses differ greatly in young and old animals. PMID- 19679151 TI - Enzymatic substrates in microbiology. AB - Enzymatic substrates are powerful tools in biochemistry. They are widely used in microbiology to study metabolic pathways, to monitor metabolism and to detect, enumerate and identify microorganisms. Synthetic enzymatic substrates have been customized for various microbial assays, to detect an expanding range of both new enzymatic activities and target microorganisms. Recent developments in synthetic enzymatic substrates with new spectral, chemical and biochemical properties allow improved detection, enumeration and identification of food-borne microorganisms, clinical pathogens and multi-resistant bacteria in various sample types. In the past 20 years, the range of synthetic enzymatic substrates used in microbiology has been markedly extended supporting the development of new multi-test systems (e.g., Microscan, Vitek 2, Phoenix) and chromogenic culture media. The use of such substrates enables an improvement in time to detection and specificity over conventional tests that employ natural substrates. In the era of intense developments in molecular biology, phenotypic tests involving enzymatic substrates remain useful to analyse both simple and complex samples. Such tests are applicable to diagnostic and research laboratories all over the world. PMID- 19679152 TI - Real-time electrochemical monitoring of drug release from therapeutic nanoparticles. AB - An electrochemical protocol for real-time monitoring of drug release kinetics from therapeutic nanoparticles (NPs) is described. The method is illustrated for repetitive square-wave voltammetric measurements of the reduction of doxorubicin released from liposomes at a glassy-carbon electrode. Such operation couples high sensitivity down to 20 nM doxorubicin with high speed and stability. It can thus monitor in real time the drug release from NP carriers, including continuous measurements in diluted serum. Such direct and continuous monitoring of the drug release kinetics from therapeutic NPs holds great promise for designing new drug delivery NPs with optimal drug release properties. These NPs can potentially be used to deliver many novel compounds such as marine-life derived drugs and hydrophobic drugs with limited water solubility that are usually difficult to be characterized by traditional analytical tools. PMID- 19679154 TI - Genetic association study of FOXP3 polymorphisms in allergic rhinitis in a Chinese population. AB - The FOXP3 gene encodes a transcription factor thought to be essential for the development and function of regulatory T cells, which are prevailing mediators of immunological tolerance via suppression/modulation of both T helper (Th)-1 and Th 2 mediated immune responses. Previous studies have demonstrated an association between common polymorphisms in FOXP3 and a number of immune diseases. The aim of this study was to investigate whether genetic polymorphisms at the FOXP3 locus predispose to allergic rhinitis (AR) in a Chinese Han population. Six polymorphisms in promoter and intron areas were genotyped in 193 AR subjects and 191 healthy controls. Twelve exons were also analyzed among cohorts of 157 AR patients and 118 healthy controls. Whole-population and gender strata analyses revealed that no single nucleotide polymorphisms in FOXP3 were identified as significantly associated with AR. Regarding the stratified analysis for heterozygotes and homozygotes, the heterozygous allele in rs3761548 (p = 0.020, OR(Het) = 3.12) appeared significant. Subgroup analysis for the presence of different allergen allergies also demonstrated a significant association for house dust mites (rs3060515, p = 0.010, odds ratio (OR) = 2.18; rs3761547, p = 0.013, OR = 2.00). Additionally, no polymorphisms in coding regions contributing to a susceptibility to AR were noted. Our study provides the first evidence for the association of the FOXP3 polymorphism with AR in a Chinese population. PMID- 19679153 TI - Removal of B cell epitopes as a practical approach for reducing the immunogenicity of foreign protein-based therapeutics. AB - Immunogenicity of non-human proteins with useful therapeutic properties has prevented their development for use in the therapy of disease. However, this class of proteins could be very useful, if their immunogenicity could be markedly reduced so that many treatment cycles could be administered. One approach to reduce the immunogenicity of foreign proteins is to identify B cell epitopes on the protein and eliminate them by mutagenesis. In this article, theoretical aspects and experimental evidence for the feasibility of B cell epitope removal is reviewed. A special focus is given to our results with deimmunization of recombinant immunotoxins in which Fvs are fused to a 38kDa portion of the bacterial protein, Pseudomonas exotoxin A (PE38). Immunotoxins targeting CD22 and CD25 have produced complete remissions in many patients with drug resistant Hairy Cell Leukemia and are being evaluated in other malignancies. Experimental data summarized in this review indicates that removal of B cell epitopes is a practical approach for making less immunogenic protein therapeutics from non human functional proteins. This approach requires grouping of the epitopes to identify targets for deimmunization followed by quantitative analysis of the decrease in affinity produced by the mutations in B cell epitopes. PMID- 19679155 TI - High levels of molecular polymorphism at the KIR2DL4 locus in French and Congolese populations: impact for anthropology and clinical studies. AB - To characterize KIR2DL4 molecular polymorphism, a cloning-sequencing protocol was performed in 49 French and 52 Teke Congolese individuals. These two populations exhibited high levels of genetic diversity for KIR2DL4, possibly under the influence of natural selection. The most frequent alleles in French individuals (i.e., *00801 and *00802 with a cumulated frequency of approximately 43%) were not the same in Congolese individuals (i.e., *00103 at 47%). In the latter population, four new allelic variants were detected, three of them harboring nonsynonymous substitutions leading to amino acid changes in the extracellular and cytoplasmic domains of the protein. Expression patterns of KIR2DL4 were tightly linked with 9 and 10 poly-adenine polymorphism in exon 7 (i.e., 9A and 10A type alleles). French individuals exhibited a majority of 9A alleles (62%), whereas Congolese individuals had a dominant subset of 10A alleles (72%), suggesting that KIR2DL4 polymorphism could be under the influence of various environmental and pathogenic backgrounds. We conclude that KIR2DL4 might be a good candidate to study for anthropology. In addition, the discovery of its intrinsic variability is shedding light on potential differences among human populations in relation to immunologic functions. PMID- 19679156 TI - Comparative study of GH-transgenic and non-transgenic amago salmon (Oncorhynchus masou ishikawae) allergenicity and proteomic analysis of amago salmon allergens. AB - Genetically modified (GM) foods are beneficial from the standpoint of ensuring a constant supply of foodstuffs, but they must be tested for safety before being released on the market, including by allergenicity tests to ensure that they do not contain new allergens or higher concentrations of known allergens than the same non-GM foods. In this study we used GM-amago salmon into which a growth hormone gene had been introduced and compared the allergens contained in the GM and the non-GM-amago salmons. We used a combination of Western blotting with allergen-specific antibodies and a proteomic analysis of their allergens with patients' sera, a so-called allergenome analysis, to analyze allergens. Western blotting with specific antibodies showed no increase in the content of the known allergens fish parvalbumin and fish type-I collagen in GM-amago salmon, in comparison with their content in non-GM-amago salmon. The allergenome analysis of two fish-allergic patients allowed us to identify several IgE-binding proteins in amago salmon, including parvalbumin, triose-phosphate isomerase, fructose bisphosphate aldolase A, and serum albumin, and there were no qualitative differences in these proteins between GM and non-GM-amago salmons. These results indicate that amago salmon endogenous allergen expression does not seem to be altered by genetic modification. PMID- 19679158 TI - Dissimilarity in the channel intrinsic stability among the bacterial KcsA and the inwardly rectifying potassium channel ROMK1. AB - In this study, we compared the channel intrinsic stability of the bacterial K(+) channel KcsA and the inwardly rectifying potassium channel (Kir) ROMK1. ROMK1 was successfully cloned, expressed and purified from Saccharomyces cerevisae. By conventional gel electrophoresis, ROMK1 was detected in monomeric form running exclusively at approximately 45 kDa either in its oxidized or reduced form. By perfluoro-octanoic acid (PFO)-PAGE, the reduced ROMK1 was identified as tetrameric as well as oligomeric complex. However, in its oxidized form ROMK1 was exclusively detected in oligomeric form thus indicating the role of intrinsic cysteine residues and formation of disulfide bonds in stabilizing the oligomeric ROMK1. On the other hand, KcsA purified from E. coli was detected as an extremely stable tetramer both in its oxidized or reduced forms as determined by conventional or PFO-PAGE. Furthermore, in planar lipid bilayer ROMK1 exhibited prominent inward rectification, low single channel conductance and high channel open probability as compared to the KcsA channel which showed typically slight outward rectification and low open probability under similar conditions. Our experiments clearly indicate that KcsA and ROMK1 channels differ with regard to their intrinsic stability which might be related to their structural and functional differences. PMID- 19679157 TI - Revisiting delta-6 desaturase regulation by C18 unsaturated fatty acids, depending on the nutritional status. AB - Dietary polyunsaturated fatty acids (PUFA) play a key role in regulating delta-6 desaturase (D6D), the key enzyme for long-chain PUFA biosynthesis. Nevertheless, the extent of their effects on this enzyme remains controversial and difficult to assess. It has been generally admitted that C18 unsaturated fatty acids (UFAs) regulate negatively delta-6 desaturase (D6D). This inhibition has been evidenced in regard to a high glucose/fat free (HG/FF) diet used in reference. However, several nutritional investigations did not evidence any inhibition of desaturases when feeding fatty acids. Because the choice of the basal diet appeared to be of primary importance in such experiments, our goal was to reconsider the specific role of dietary UFAs on D6D regulation, depending on nutritional conditions. For that, sixteen adult Wistar rats were fed purified linoleic acid, alpha-linolenic acid or oleic acid, included in one of two diets at 4% by weight: an HG/FF or a high starch base (HS) where the pure UFAs replaced a mixed vegetable oil. Our results showed first that D6D specific activity was significantly greater when measured in presence of an HG/FF than with an HS/4% vegetable oil diet. Secondly, we found that linoleic and alpha-linolenic acids added to HG/FF reduced the specific activity of D6D. In contrast, when pure UFAs were added to an HS base, D6D specific activities remained unchanged or increased. Concordant results were obtained on D6D mRNA expression. Altogether, this study evidenced the importance of the nutritional status in D6D regulation by C18 UFAs: when used as control, HG/FF diet stimulates D6D compared with a standard control diet containing starch and 4% fats, leading to an overestimation of the D6D regulation by UFAs. Then, UFAs should be considered as repressors for unsaturated fatty acid biosynthesis only in very specific nutritional conditions. PMID- 19679159 TI - Plasma fatty acid levels may regulate the Zn(2+)-dependent activities of histidine-rich glycoprotein. AB - Histidine-rich glycoprotein (HRG) is a plasma adaptor protein involved in the formation of protein complexes that regulate a number of biological processes in the blood, most notably coagulation and the immune system. Elevated levels of HRG are clinically linked to thrombotic disorders such as blood vessel occlusion. A large body of evidence suggests that Zn(2+) ions stimulate HRG-complex formation; however, under normal conditions the vast majority of Zn(2+) in the blood is bound to human serum albumin (HSA). We have previously demonstrated that high levels of fatty acid act as an allosteric switch which disrupts the major Zn(2+) binding site on HSA. Transient or sustained elevation of plasma fatty acid levels may therefore increase the proportion of plasma Zn(2+) associated with HRG. We speculate that this mechanism may potentiate an increased risk of thrombosis in individuals with elevated fatty acid levels such as those associated with cancer, obesity and diabetes. PMID- 19679160 TI - Characterization of papain-like isoenzymes from latex of Asclepias curassavica by molecular biology validated by proteomic approach. AB - Latices from Asclepias spp are used in wound healing and the treatment of some digestive disorders. These pharmacological actions have been attributed to the presence of cysteine proteases in these milky latices. Asclepias curassavica (Asclepiadaceae), "scarlet milkweed" is a perennial subshrub native to South America. In the current paper we report a new approach directed at the selective biochemical and molecular characterization of asclepain cI (acI) and asclepain cII (acII), the enzymes responsible for the proteolytic activity of the scarlet milkweed latex. SDS-PAGE spots of both purified peptidases were digested with trypsin and Peptide Mass Fingerprints (PMFs) obtained showed no equivalent peptides. No identification was possible by MASCOT search due to the paucity of information concerning Asclepiadaceae latex cysteine proteinases available in databases. From total RNA extracted from latex samples, cDNA of both peptidases was obtained by RT-PCR using degenerate primers encoding Asclepiadaceae cysteine peptidase conserved domains. Theoretical PMFs of partial polypeptide sequences obtained by cloning (186 and 185 amino acids) were compared with empirical PMFs, confirming that the sequences of 186 and 185 amino acids correspond to acI and acII, respectively. N-terminal sequences of acI and acII, characterized by Edman sequencing, were overlapped with those coming from the cDNA to obtain the full length sequence of both mature peptidases (212 and 211 residues respectively). Alignment and phylogenetic analysis confirmed that acI and acII belong to the subfamily C1A forming a new group of papain-like cysteine peptidases together with asclepain f from Asclepias fruticosa. We conclude that PMF could be adopted as an excellent tool to differentiate, in a fast and unequivocal way, peptidases with very similar physicochemical and functional properties, with advantages over other conventional methods (for instance enzyme kinetics) that are time consuming and afford less reliable results. PMID- 19679161 TI - An evolutionary system using development and artificial Genetic Regulatory Networks for electronic circuit design. AB - Biology presents incomparable, but desirable, characteristics compared to engineered systems. Inspired by biological development, we have devised a multi layered design architecture that attempts to capture the favourable characteristics of biological mechanisms for application to design problems. We have identified and implemented essential features of Genetic Regulatory Networks (GRNs) and cell signalling which lead to self-organization and cell differentiation. We have applied this to electronic circuit design. PMID- 19679162 TI - Implicit association between basic tastes and pitch. AB - Our evaluation of food is influenced by a variety of contextual information perceived via the senses. Here we investigated whether there are interactions between auditory stimuli and basic tastes (indicated by the names of typically sour and bitter foods). Participants took part in a version of the Implicit Association Test (IAT) in order to measure the strength of the association between high-pitched sounds and (the names of) foodstuffs having a sour taste, and between low-pitched sounds and (the names of) foodstuffs having a bitter taste. Analysis of the latency and accuracy of participants' responses highlighted the existence of a significant crossmodal association between these different attributes. This result suggests the need for research into the influence of auditory stimuli on food evaluation, an interaction that has typically been overlooked when considering the multisensory perception of food. PMID- 19679163 TI - Regulation of non-coding RNA networks in the nervous system--what's the REST of the story? AB - Recent advances are now providing novel insights into the mechanisms that underlie how cellular complexity, diversity, and connectivity are encoded within the genome. The repressor element-1 silencing transcription factor/neuron restrictive silencing factor (REST/NRSF) and non-coding RNAs (ncRNAs) are emerging as key regulators that seem to orchestrate almost every aspect of nervous system development, homeostasis, and plasticity. REST and its primary cofactor, CoREST, dynamically recruit highly malleable macromolecular complexes to widely distributed genomic regulatory sequences, including the repressor element-1/neuron restrictive silencer element (RE1/NRSE). Through epigenetic mechanisms, such as site-specific targeting and higher-order chromatin remodeling, REST and CoREST can mediate cell type- and developmental stage specific gene repression, gene activation, and long-term gene silencing for protein-coding genes and for several classes of ncRNAs (e.g. microRNAs [miRNAs] and long ncRNAs). In turn, these ncRNAs have similarly been implicated in the regulation of chromatin architecture and dynamics, transcription, post transcriptional processing, and RNA editing and trafficking. In addition, REST and CoREST expression and function are tightly regulated by context-specific transcriptional and post-transcriptional mechanisms including bidirectional feedback loops with various ncRNAs. Not surprisingly, deregulation of REST and ncRNAs are both implicated in the molecular pathophysiology underlying diverse disorders that range from brain cancer and stroke to neurodevelopmental and neurodegenerative diseases. This review summarizes emerging aspects of the complex mechanistic relationships between these intricately interlaced control systems for neural gene expression and function. PMID- 19679165 TI - In vivo down-regulation of mouse brain capillary P-glycoprotein: a preliminary investigation. AB - Over-expression of blood-brain barrier P-glycoprotein is considered as a major hurdle in the treatment of various CNS disorders. A down-regulation strategy is considered as one means to counteract disease- or therapy-associated induction of P-glycoprotein. Here, we evaluated whether a targeting of P-glycoprotein can be achieved in mouse brain capillary endothelial cells using siRNA. A 4-day treatment paradigm with once daily hydrodynamic intravenous injections of siRNA resulted in a significant reduction of the P-glycoprotein-labeled area in the hippocampal hilus and parietal cortex. P-glycoprotein expression proved to be down-regulated in these brain regions by 31 and 16%, respectively. An impact of siRNA administration on density of brain capillaries was excluded by quantification of the endothelial cell marker GLUT-1. In conclusion, the study provides first preliminary evidence that a down-regulation of P-glycoprotein can be achieved in brain capillary endothelial cells by administration of siRNA in vivo. PMID- 19679164 TI - Regulation of Fcgamma receptors and immunoglobulin G-mediated phagocytosis in mouse microglia. AB - As resident macrophages in the CNS, microglia can transform from a surveillance state to an activated phenotype in response to brain injury. During this transition microglia become highly capable phagocytic cells. Invading pathogens undergo opsonization with immunoglobulins and microglia recognize these opsonized pathogens through interaction with their cognate F(c) receptors. In mice, both FcgammaRI and FcgammaRIIb receptors are involved in IgG-mediated phagocytosis of opsonzied pathogens. At sites of inflammation, microglial activity is regulated by T-cell derived cytokines. Here we first investigated the effects of IFN-gamma, IL-4, IL-13 and GM-CSF on expression of FcgammaRI and FcgammaRIIb mRNA levels in both primary microglia and microglial cell line N9. Using quantitative real-time PCR we show that IFN-gamma induced a 4-fold increase in the mRNA level of FcgammaRI but did not induce changes in FcgammaRIIb expression. IL-4 and IL-13 induced approximately 2-fold increases in expression of FcgammaRIIb mRNA, but had no effect on FcgammaRI expression. GM-CSF increased both FcgammaRI and FcgammaRIIb mRNA expression. We then characterized the ability of these same cytokines to regulate phagocytosis of immune complexes composed of IgG and the bacteria Staphylococcus aureus. IFN-gamma and GM-CSF both induced approximately 2 fold increases in IgG-mediated phagocytosis whereas IL-4 and IL-13 both decreased IgG-mediated phagocytosis by about one-third. None of the cytokines influenced basal levels of phagocytosis. These findings demonstrate a highly selective cytokine-induced regulation of both phagocytosis-related Fcgamma receptor subtypes and IgG-mediated phagocytosis itself in microglia. This selective regulation has implications for our understanding of the pathophysiology of CNS infection and autoimmune disease. PMID- 19679166 TI - TPH2 polymorphisms may modify clinical picture in treatment-resistant depression. AB - The association of two tryptophan hydroxylase 2 (TPH2) polymorphisms and treatment response in electroconvulsive therapy (ECT) and the risk of depression was studied. The patient sample consisted of 119 subjects with treatment resistant major depressive disorder who were treated with ECT. Treatment response was assessed by the Montgomery and Asberg Depression Rating Scale (MADRS) scores. Patients who had <8 scores in post-treatment MADRS were considered remitters; scores >15 indicated non-response. The polymorphisms studied (rs1386494 and rs1843809) were not associated with treatment response to ECT. However, TPH2 rs1386494 A/A genotype carrying patients had significantly higher MADRS scores before ECT than A/G+G/G genotype carriers (p<0.001). A/A genotype carriers also had a greater decline in MADRS scores than A/G+G/G genotype carriers during the course of ECT treatment (p=0.03). This polymorphism may be associated with the severity of treatment-resistant depression. ECT may able to counteract a putative genetically driven worse depressive phenotype. PMID- 19679167 TI - Involvement of nitric oxide synthesis in sensitization to the rewarding effects of morphine. AB - Knowledge about the specific brain changes and neural plasticity processes produced by repeated exposure to a drug is essential to progress in the field of neurobiology of addiction and the development of effective medication. In the present study, the influence of nitric oxide synthesis on sensitization to the rewarding effects of morphine has been evaluated. The effects of pre-treatment of mice with saline or 20mg/kg of morphine plus the nitric oxide synthase inhibitor 7-nitroindazole (7NI) (12.5 or 25mg/kg) on the place conditioning induced by a low dose of morphine (2mg/kg) were assessed. The dose of 2mg/kg of morphine was ineffective in animals pre-treated with saline but induced a clear conditioned place preference (CPP) in those pre-treated with morphine. Conversely, animals pre-treated with morphine plus 7NI did not acquire CPP. Our results demonstrate that the nitric oxide pathway is implicated in the development of sensitization to the conditioned rewarding effects of morphine. PMID- 19679168 TI - A monoclonal antibody as a tool to study the subcommissural organ and Reissner's fibre of the sea lamprey: an immunofluorescence study before and after a spinal cord transection. AB - Lampreys are vertebrate animal models in spinal cord regeneration studies. In order to gain knowledge on the mechanisms that provide to the lamprey spinal cord its capacity of regeneration we decided to compare the expression patterns of the growth-associated protein 43 (GAP-43) in the CNS of the sea lamprey before and after a complete spinal transection by immunocytochemical methods using an anti GAP-43 antibody. Surprisingly, in the brain/spinal cord of both normal and injured animals, anti-GAP-43-like labeling was only observed in the subcommissural organ (SCO) and Reissner's fibre (RF). In injured larvae, a dotted labeling was also observed in the meninges and in the blood the vessels of the neighbouring tissues at the site of lesion. The experiments in injured animals showed that after complete spinal cord transection the SCO seems to continue to produce the Reissner's substance (RS), which is accumulated at the proximal site of spinal transection. The dotted labeling observed in the neighbouring tissues could correspond to RS that was released from the site of injury. In Western blot experiments done using protein extracts of the lamprey brain, the anti-GAP-43 antibody did not recognize any protein band of the expected GAP-43 molecular weight, indicating that the secreted material is not this protein. An anti serotonin antibody was also used as a marker of some brain structures. Serotonergic afferent fibres innervated the SCO. Here we show a new tool that can be used as a highly specific marker in further studies of the SCO/RF system of lampreys. PMID- 19679169 TI - Age-related changes in antioxidant enzymes related to hydrogen peroxide metabolism in rat inner ear. AB - Oxidative stress is a pervasive factor in aging and has been implicated in noise induced cochlear pathology. In this study, we measured the activities of two enzymes that catalyze the removal of hydrogen peroxide (H(2)O(2)), catalase and glutathione peroxidase (Gpx), in 3- and 24-month-old Fisher-344 rats, and reduced and oxidized glutathione in 3-, 12-, and 24-month-old rats. There was an increase in Gpx activity in vascular tissue (spiral ligament and stria vascularis), but no change in modiolar, sensory or vestibular tissue of the cochlea. The elevation in vascular tissue was age-related. We observed a significant elevation of catalase activity in vestibular tissue, a tendency for age-related elevation in the modiolus, but no change in vascular or sensory cochlear tissue. These findings suggest that increased Gpx activity in vascular cochlear tissue may be an age related compensation for a decrease in glutathione and a decline in the redox state measured by the ratio of reduced to oxidized glutathione. PMID- 19679170 TI - Isoflurane preconditioning improves short-term and long-term neurological outcome after focal brain ischemia in adult rats. AB - Isoflurane preconditioning improved short-term neurological outcome after focal brain ischemia in adult rats. It is not known whether desflurane induces a delayed phase of preconditioning in the brain and whether isoflurane preconditioning-induced neuroprotection is long-lasting. Two months-old Sprague Dawley male rats were exposed to or were not exposed to isoflurane or desflurane for 30 min and then subjected to a 90 min middle cerebral arterial occlusion (MCAO) at 24 h after the anesthetic exposure. Neurological outcome was evaluated at 24 h or 4 weeks after the MCAO. The density of the terminal deoxynucleotidyl transferase biotinylated UTP nick end labeling (TUNEL) positive cells in the penumbral cerebral cortex were assessed 4 weeks after the MCAO. Also, rats were pretreated with isoflurane or desflurane for 30 min. Their cerebral cortices were harvested for quantifying B-cell lymphoma-2 (Bcl-2) expression 24 h later. Here, we showed that pretreatment with 1.1% or 2.2% isoflurane, but not with 6% or 12% desflurane, increased Bcl-2 expression in the cerebral cortex, improved neurological functions and reduced infarct volumes evaluated at 24 h after the MCAO. Isoflurane preconditioning also improved neurological functions and reduced brain infarct volumes in rats evaluated 4 weeks after the MCAO. Isoflurane preconditioning also decreased the density of TUNEL-positive cells in the penumbral cerebral cortex. We conclude that isoflurane preconditioning improves short-term and long-term neurological outcome and reduces delayed cell death after transient focal brain ischemia in adult rats. Bcl-2 may be involved in the isoflurane preconditioning effect. Desflurane pretreatment did not induce a delayed phase of neuroprotection. PMID- 19679171 TI - Peripheral estradiol induces temporomandibular joint antinociception in rats by activating the nitric oxide/cyclic guanosine monophosphate signaling pathway. AB - Recently, we have reported that high physiological estradiol level during the proestrus phase of the estrous cycle or systemic estradiol administration in ovariectomized rats decreases formalin-induced temporomandibular joint nociception. However, the mechanisms underlying the antinociceptive effect of estradiol are presently unknown. In this study, we used the temporomandibular joint formalin model in rats to investigate whether estradiol decreases nociception by a peripheral non-genomic mechanism, and if so, whether this mechanism is mediated by the activation of the nitric oxide-cyclic guanosine monophosphate signaling pathway and of opioid receptors. The administration of estradiol into the ipsilateral, but not into the contralateral temporomandibular joint significantly reduced formalin-induced temporomandibular joint nociception in ovariectomized and diestrus but not in proestrus females. However, the administration of the estrogen receptor antagonist ICI 182780 into the ipsilateral, but not into the contralateral temporomandibular joint blocked the antinociceptive effect of serum estradiol in proestrus females, suggesting that the physiological effect of estradiol in nociception is mediated, at least in part, by a peripheral mechanism. The administration of estradiol into the ipisilateral temporomandibular joint did not affect formalin-induced nociception in male rats. The antinociceptive effect of temporomandibular joint estradiol administration in ovariectomized and diestrus females was mimicked by estradiol conjugated with bovine serum albumin, which does not diffuse through the plasma membrane, and was blocked by the estrogen receptor antagonist ICI 182780. The administration of the nitric oxide synthase inhibitor (nitro-l-arginine) or of a guanylate cyclase inhibitor (1H-(1,2,4)-oxadiasolo (4,2-a) quinoxalin-1-one) into the ipsilateral, but not into the contralateral temporomandibular joint blocked the antinociceptive effect of estradiol and of estradiol conjugated with bovine serum albumin, while the opioid receptor antagonist naloxone had no effect. These findings suggest that estradiol decreases temporomandibular joint nociception in female rats through a peripheral non-genomic activation of the nitric oxide cyclic guanosine monophosphate signaling pathway. PMID- 19679172 TI - Brown adipose tissue thermogenesis heats brain and body as part of the brain coordinated ultradian basic rest-activity cycle. AB - Brown adipose tissue (BAT), body and brain temperatures, as well as behavioral activity, arterial pressure and heart rate, increase episodically during the waking (dark) phase of the circadian cycle in rats. Phase-linking of combinations of these ultradian (<24 h) events has previously been noted, but no synthesis of their overall interrelationships has emerged. We hypothesized that they are coordinated by brain central command, and that BAT thermogenesis, itself controlled by the brain, contributes to increases in brain and body temperature. We used chronically implanted instruments to measure combinations of bat, brain and body temperatures, behavioral activity, tail artery blood flow, and arterial pressure and heart rate, in conscious freely moving Sprague-Dawley rats during the 12-h dark active period. Ambient temperature was kept constant for any particular 24-h day, varying between 22 and 27 degrees C on different days. Increases in BAT temperature (> or = 0.5 degrees C) occurred in an irregular episodic manner every 94+/-43 min (mean+/-SD). Varying the temperature over a wider range (18-30 degrees C) on different days did not change the periodicity, and neither body nor brain temperature fell before BAT temperature episodic increases. These increases are thus unlikely to reflect thermoregulatory homeostasis. Episodic BAT thermogenesis still occurred in food-deprived rats. Behavioral activity, arterial pressure (18+/-5 mmHg every 98+/-49 min) and heart rate (86+/-31 beats/min) increased approximately 3 min before each increase in BAT temperature. Increases in BAT temperature (1.1+/-0.4 degrees C) were larger than corresponding increases in brain (0.8+/-0.4 degrees C) and body (0.6+/-0.3 degrees C) temperature and the BAT episodes commenced 2-3 min before body and brain episodes, suggesting that BAT thermogenesis warms body and brain. Hippocampal 5-8 Hz theta rhythm, indicating active engagement with the environment, increased before the behavioral and autonomic events, suggesting coordination by brain central command as part of the 1-2 h ultradian basic rest activity cycle (BRAC) proposed by Kleitman. PMID- 19679173 TI - Cerebral metabolism and sleep homeostasis: a comment on Vyazovskiy et al. AB - In this journal, Vyazovskiy et al. reported deoxyglucose data that they interpreted as supporting the Tononi and Cirelli hypothesis that cerebral metabolic rate (CMR) increases across waking. We summarize contradictory PET deoxyglucose data that show CMR is the same in the morning and evening and that it decreases rather than increases after a night of sleep deprivation. We propose an alternative model of CMR-sleep homeostasis relations that points to novel experiments. PMID- 19679174 TI - Targeting retinal and choroid neovascularization using the small molecule inhibitor carboxyamidotriazole. AB - Neovascular ocular diseases as exemplified by proliferative diabetic retinopathy (PDR), exudative age-related macular degeneration (AMD), and retinopathy of prematurity (ROP) are severe diseases affecting all age groups in the US. We asked whether a small molecule, carboxyamidotriazole (CAI) known for its anti angiogenic and anti-tumor effects and its ability to be administered orally in humans, could have anti-angiogenic effects in ocular in vitro and in vivo angiogenesis models. The anti-proliferative effects of CAI were examined by BrdU incorporation using human retinal and dermal endothelial cells and human pigment epithelial cells. The effect of CAI was determined using the Matrigel tube formation assay. The mouse model of choroidal neovascularization (CNV) initiated by laser rupture of Bruch's membrane was used to quantify in vivo effects of aqueous beta-hydroxypropyl cyclodextrin (bHPCD) formulations of CAI on neovascularization. The pharmacokinetics (PK) of CAI after intravitreal administration of bHPCD-CAI was studied in rabbit. The intravitreal toxicology of bHPCD-CAI was also examined in rat ocular tissue. We observed that CAI treatment of human endothelial cells decreased cell proliferation in a dose-dependent manner. In the in vivo tests bHPCD-CAI treatment reduced choroidal neovascular lesion volume, also in a dose-dependent manner. The intravitreal PK of bHPCD-CAI demonstrated that highly efficacious concentrations of CAI are reached in the vitreous compartment. No ocular toxicology was observed with intravitreous injection of CAI. These studies support the potential of developing intravitreal CAI in an bHPCD ocular formulation for treatment of proliferative retinopathies in humans. PMID- 19679175 TI - Challenges in Nano- Micro- and Macro-systems. Preface. PMID- 19679176 TI - In vitro and in vivo anti-photoaging effects of an isoflavone extract from soybean cake. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Soy has been used in traditional Chinese medicine for thousands of years for its health and nutritional benefits, as well as to treat and care for the skin. Advanced skin care research has shown that soy isoflavone and genistein are effective in reducing damage to the skin from the sun. AIM OF THE STUDY: To study the protective effects of isoflavone extract from soybean cake against the UVB-induced skin damage. MATERIALS AND METHODS: The in vitro effects and possible mechanisms of soybean extract on UVB protection were determined in HaCaT cells. In the in vivo study, ICR-Foxn/(nu) mice were irradiated with UVB. The epidermal thickness, catalase and the expressions of cyclooxygenase-2 (COX-2) and proliferating cell nuclear antigen (PCNA) were detected to evaluate the antioxidant and anti-inflammatory activities of the isoflavone extract. RESULTS: Our in vitro studies showed that UVB-induced HaCaT cell death and the phosphorylation of p38, JNK, and ERK1/2 decreased in the presence of isoflavone extract. In the in vivo studies, we found that the topical application of isoflavone extract before UVB irritation decreased the epidermal thickness and the expressions of COX-2 and PCNA and increased catalase concentration. These results showed anti-photoaging effect of isoflavone extract from soybean cake involved the inhibition of UVB-induced apoptosis and inflammation. CONCLUSIONS: It is shown that isoflavone extract from soybean cake may be functional cosmeceutical candidate for skin photoaging. PMID- 19679177 TI - Evolutionary genomics of the Fox genes: origin of gene families and the ancestry of gene clusters. AB - Over the past decade genomic approaches have begun to revolutionise the study of animal diversity. In particular, genome sequencing programmes have spread beyond the traditional model species to encompass an increasing diversity of animals from many different phyla, as well as unicellular eukaryotes that are closely related to the animals. Whole genome sequences allow researchers to establish, with reasonable confidence, the full complement of any particular family of genes in a genome. Comparison of gene complements from appropriate genomes can reveal the evolutionary history of gene families, indicating when both gene diversification and gene loss have occurred. More than that, however, assembled genomes allow the genomic environment in which individual genes are found to be analysed and compared between species. This can reveal how gene diversification occurred. Here, we focus on the Fox genes, drawing from multiple animal genomes to develop an evolutionary framework explaining the timing and mechanism of origin of the diversity of animal Fox genes. Ancient linkages between genes are a prominent feature of the Fox genes, depicting a history of gene clusters, some of which may be relevant to understanding Fox gene function. PMID- 19679178 TI - Pleiotropic effects of RecQ in Deinococcus radiodurans. AB - In Deinococcus radiodurans, there is a unique RecQ homolog (DR1289) with three tandem HRDC domains. Deletion of drrecQ resulted in a low doubling rate and sensitivity to hydrogen peroxide. Here, we used cDNA microarray and biochemical assays to explore the physiological changes in the drrecQ mutant. The expressions of genes with predicted functions involved in iron homeostasis, antioxidant system, electron transport, and energy metabolism were significantly altered in response to drrecQ disruption. More reactive oxygen species (ROS) was accumulated in drrecQ mutant strain when compared to wild type. In addition, ICP-MS results showed that the intracellular level of iron was relatively higher, whereas the concentration of manganese was lower in drrecQ mutant than in wild type. Furthermore, our microarray data and pulsed-field gel results showed that DNA suffered more damage in drrecQ mutant than in wild type under 20 mM hydrogen peroxide stress. These results suggested that drrecQ is a gene of pleiotropic functions and contributes to the extraordinary resistance of D. radiodurans against stresses. PMID- 19679179 TI - Why should an immune response activate the stress response? Insights from the insects (the cricket Gryllus texensis). AB - Mediators of the stress response (e.g. glucocorticoids and norepinephrine) can be immunosuppressive. Nevertheless, immune challenge leads to the release of these compounds in vertebrates. To resolve this paradox, it has been suggested that stress hormones help restore immune homeostasis, preventing self-damage. A comparative approach may provide additional hypotheses as to why an immune challenge induces the release of stress hormones/neurohormones. Octopamine, a neurohormonal mediator of the stress response in the cricket Gryllus texensis, increased in concentration in the hemolymph during an immune challenge. Therefore, the release of stress hormones during an immune response occurs in animals across phyla. Octopamine induced an increase in lipid concentration in the hemolymph. After an acute stress (flying or running) the total number of hemocytes in the hemolymph increased. Injections of octopamine had the same effect, suggesting that it may enhance hemocyte-dependent immune functions. On the other hand, octopamine decreased lysozyme-like activity in vitro, suggesting that it inhibits some immune functions. However, lysozyme-like activity was increased by the presence of heat-killed bacteria in vitro and this increase was significantly augmented by the presence of octopamine. Therefore, the effect of octopamine on immune function differed depending on the presence of pathogens. Stress hormones may help shift immune function into the most optimal configuration depending on the physiological context. PMID- 19679180 TI - Glial cell line-derived neurotrophic factor acutely modulates the excitability of rat small-diameter trigeminal ganglion neurons innervating facial skin. AB - Glial cell line-derived neurotrophic factor (GDNF) plays an important role in adult sensory neuron function. However, the acute effects of GDNF on primary sensory neuron excitability remain to be elucidated. The aim of the present study was to investigate whether GDNF acutely modulates the excitability of adult rat trigeminal ganglion (TRG) neurons that innervate the facial skin by using perforated-patch clamping, retrograde-labeling and immunohistochemistry techniques. Fluorogold (FG) retrograde labeling was used to identify the TRG neurons innervating the facial skin. The FG-labeled small- and medium-diameter GDNF immunoreactive TRG neurons, and most of these neurons also expressed the GDNF family receptor alpha-1 (GFRalpha-1). In whole-cell voltage-clamp mode, GDNF application significantly inhibited voltage-gated K(+) transient (I(A)) and sustained (I(K)) currents in most dissociated FG-labeled small-diameter TRG neurons. This effect was concentration-dependent and was abolished by co application of the protein tyrosine kinase inhibitor, K252b. Under current-clamp conditions, the repetitive firing during a depolarizing pulse were significantly increased by GDNF application. GDNF application also increased the duration of the repolarization phase and decreased the duration of the depolarization phase of the action potential, and these characteristic effects were also abolished by co-application of K252b. These results suggest that acute application of GDNF enhances the neuronal excitability of adult rat small-diameter TRG neurons innervating the facial skin, via activation of GDNF-induced intracellular signaling pathway. We therefore conclude that a local release of GDNF from TRG neuronal soma and/or nerve terminals may regulate normal sensory function, including nociception. PMID- 19679181 TI - Evidence that opioids may have toll-like receptor 4 and MD-2 effects. AB - Opioid-induced proinflammatory glial activation modulates wide-ranging aspects of opioid pharmacology including: opposition of acute and chronic opioid analgesia, opioid analgesic tolerance, opioid-induced hyperalgesia, development of opioid dependence, opioid reward, and opioid respiratory depression. However, the mechanism(s) contributing to opioid-induced proinflammatory actions remains unresolved. The potential involvement of toll-like receptor 4 (TLR4) was examined using in vitro, in vivo, and in silico techniques. Morphine non-stereoselectively induced TLR4 signaling in vitro, blocked by a classical TLR4 antagonist and non stereoselectively by naloxone. Pharmacological blockade of TLR4 signaling in vivo potentiated acute intrathecal morphine analgesia, attenuated development of analgesic tolerance, hyperalgesia, and opioid withdrawal behaviors. TLR4 opposition to opioid actions was supported by morphine treatment of TLR4 knockout mice, which revealed a significant threefold leftward shift in the analgesia dose response function, versus wildtype mice. A range of structurally diverse clinically-employed opioid analgesics was found to be capable of activating TLR4 signaling in vitro. Selectivity in the response was identified since morphine-3 glucuronide, a morphine metabolite with no opioid receptor activity, displayed significant TLR4 activity, whilst the opioid receptor active metabolite, morphine 6-glucuronide, was devoid of such properties. In silico docking simulations revealed ligands bound preferentially to the LPS binding pocket of MD-2 rather than TLR4. An in silico to in vitro prediction model was built and tested with substantial accuracy. These data provide evidence that select opioids may non stereoselectively influence TLR4 signaling and have behavioral consequences resulting, in part, via TLR4 signaling. PMID- 19679183 TI - Approaching Drosophila development through proteomic tools and databases: At the hub of the post-genomic era. AB - The past decade has witnessed an explosion in the growth of proteomics. The completion of numerous genome sequences, the development of powerful protein analytical technologies, as well as the design of innovative bioinformatics tools have marked the beginning of a new post-genomic era. Proteomics, the large-scale analysis of proteins in an organism, organ or organelle encompasses different aspects: (1) the identification, analysis of post-translational modifications and quantification of proteins; (2) the study of protein-protein interactions; and (3) the functional analysis of interactome networks. Here, we briefly summarize the emerging analytical tools and databases that are paving the way for studying Drosophila development by proteomic approaches. PMID- 19679182 TI - Frataxin deficiency induces Schwann cell inflammation and death. AB - Mutations in the frataxin gene cause dorsal root ganglion demyelination and neurodegeneration, which leads to Friedreich's ataxia. However the consequences of frataxin depletion have not been measured in dorsal root ganglia or Schwann cells. We knocked down frataxin in several neural cell lines, including two dorsal root ganglia neural lines, 2 neuronal lines, a human oligodendroglial line (HOG) and multiple Schwann cell lines and measured cell death and proliferation. Only Schwann cells demonstrated a significant decrease in viability. In addition to the death of Schwann cells, frataxin decreased proliferation in Schwann, oligodendroglia, and slightly in one neural cell line. Thus the most severe effects of frataxin deficiency were on Schwann cells, which enwrap dorsal root ganglia neurons. Microarray of frataxin-deficient Schwann cells demonstrated strong activations of inflammatory and cell death genes including interleukin-6 and Tumor Necrosis Factor which were confirmed at the mRNA and protein levels. Frataxin knockdown in Schwann cells also specifically induced inflammatory arachidonate metabolites. Anti-inflammatory and anti-apoptotic drugs significantly rescued frataxin-dependent Schwann cell toxicity. Thus, frataxin deficiency triggers inflammatory changes and death of Schwann cells that is inhibitable by inflammatory and anti-apoptotic drugs. PMID- 19679184 TI - Effects of liquisolid formulations on dissolution of naproxen. AB - The aim of this study was to investigate the use of liquisolid technique in improving the dissolution profiles of naproxen in a solid dosage form. This study was designed to evaluate the effects of different formulation variables, i.e. type of non-volatile liquid vehicles and drug concentrations, on drug dissolution rates. The liquisolid tablets were formulated with three different liquid vehicles, namely Cremophor EL (polyoxyl 35 castor oil), Synperonic PE/L61 (poloxamer 181, polyoxyethylene-polyoxypropylene copolymer) and poly ethylene glycol 400 (PEG400) at two drug concentrations, 20%w/w and 40%w/w. Avicel PH102 was used as a carrier material, Cab-o-sil M-5 as a coating material and maize starch as a disintegrant. The empirical method as introduced by Spireas and Bolton (1999) [1] was applied strictly to calculate the amounts of coating and carrier materials required to prepare naproxen liquisolid tablets. Quality control tests, i.e. uniformity of tablet weight, uniformity of drug content, tablet hardness, friability test, disintegration and dissolution tests were performed to evaluate each batch of prepared tablets. In vitro drug dissolution profiles of the liquisolid formulations were studied and compared with conventional formulation, in simulated gastric fluid (pH 1.2) and simulated intestinal fluid (pH 7.2) without enzyme. Stability studies were carried out to evaluate the stability of the tablets under humid conditions. Differential scanning calorimetry and Fourier transform infrared were used to investigate physicochemical interaction between naproxen and the excipients. It was found that liquisolid tablets formulated with Cremophor EL at drug concentration of 20%w/w produced high dissolution profile with acceptable tablet properties. The stability studies showed that the dissolution profiles of liquisolid tablets prepared with Cremophor EL were not affected by ageing significantly. Furthermore, DSC revealed that drug particles in liquisolid formulations were completely solubilised. PMID- 19679185 TI - Evolutionary relationships between Fusarium oxysporum f. sp. lycopersici and F. oxysporum f. sp. radicis-lycopersici isolates inferred from mating type, elongation factor-1alpha and exopolygalacturonase sequences. AB - Fusarium oxysporum is a ubiquitous species complex of soilborne plant pathogens that comprises many different formae speciales, each characterized by a high degree of host specificity. In this study, the evolutionary relationships between different isolates of the F. oxysporum species complex have been examined, with a special emphasis on the formae speciales lycopersici and radicis-lycopersici, sharing tomato as host while causing different symptoms. Phylogenetic analyses of partial sequences of a housekeeping gene, the elongation factor-1alpha (EF 1alpha) gene, and a gene encoding a pathogenicity trait, the exopolygalacturonase (pgx4) gene, were conducted on a worldwide collection of F. oxysporum strains representing the most frequently observed vegetative compatibility groups of these formae speciales. Based on the reconstructed phylogenies, multiple evolutionary lineages were found for both formae speciales. However, different tree topologies and statistical parameters were obtained for the cladograms as several strains switched from one cluster to another depending on the locus that was used to infer the phylogeny. In addition, mating type analysis showed a mixed distribution of the MAT1-1 and MAT1-2 alleles in the F. oxysporum species complex, irrespective of the geographic origin of the tested isolates. This observation, as well as the topological conflicts that were detected between EF 1alpha and pgx4, are discussed in relation to the evolutionary history of the F. oxysporum species complex. PMID- 19679186 TI - A bicistronic expression strategy for large scale expression and purification of full-length recombinant human parathyroid hormone for osteoporosis therapy. AB - Parathyroid hormone (PTH) contributes to the increase of trabecular connectivity and is a candidate medication for effective treating osteoporosis. PTH is a protein of 84 amino acids and some studies have suggested that the active site lies within the range from amino acid (aa) 1 to 34. However, a few reports have indicated a causal relationship between PTH (aa 1-34) and osteogenic sarcoma in rats, while some less obvious but important roles of the carboxyl-terminus of PTH were also found. Unfortunately, it is difficult to obtain the active integrated PTH (1-84) in vitro, due to the instability of both the protein and its mRNA. Because an alternative translation start site is located at +25 nucleotides downstream of the true start site, a truncated PTH can be translated. We constructed a rhPTH bicistronic expression plasmid (pTrepth) that could highly express non-fusion soluble rhPTH proteins in Escherichia coli. The BL-21(DE3) containing pTrepth was cultured on a small scale until satisfactory expression and purification results were obtained. We then amplified the transformed cells in a 15-L fermentor and harvested 27g/L cells (wet weight). Extensive rhPTH purification was achieved by a three step chromatography process. Activity tests demonstrated that our purified protein could dramatically increase cAMP in osteosarcoma cells in vitro. PMID- 19679187 TI - Secretory expression and characterization of a highly Ca2+-activated thermostable L2 lipase. AB - Thermostable lipases are important biocatalysts, showing many interesting properties with industrial applications. Previously, a thermophilic Bacillus sp. strain L2 that produces a thermostable lipase was isolated. In this study, the gene encoding for mature thermostable L2 lipase was cloned into a Pichia pastoris expression vector. Under the control of the methanol-inducible alcohol oxidase (AOX) promoter, the recombinant L2 lipase was secreted into the culture medium driven by the Saccharomyces cerevisiae alpha-factor signal sequence. After optimization the maximum recombinant lipase activity achieved in shake flasks was 125 U/ml. The recombinant 44.5 kDa L2 lipase was purified 1.8-fold using affinity chromatography with 63.2% yield and a specific activity of 458.1 U/mg. Its activity was maximal at 70 degrees C and pH 8.0. Lipase activity increased 5-fold in the presence of Ca2+. L2 lipase showed a preference for medium to long chain triacylglycerols (C(10)-C(16)), corn oil, olive oil, soybean oil, and palm oil. Stabilization at high temperature and alkaline pH as well as its broad substrate specificity offer great potential for application in various industries that require high temperature operations. PMID- 19679188 TI - Expression and purification of his-tagged recombinant mouse zeta-crystallin. AB - Zeta-crystallin is an NADPH-binding protein consisting of four identical 35kD subunits. The protein possesses quinone oxidoreductase activity, and is present in large amounts in the lenses of camelids, certain hystricomorphic rodents, and the Japanese tree frog, and in lower catalytic amounts in certain tissues of various species. In this study, recombinant methods were used to produce substantial quantities of his-tagged recombinant mouse zeta-crystallin, which was then purified to homogeneity. The yield of pure recombinant mouse zeta-crystallin was five times that obtained previously for purification of recombinant guinea pig zeta-crystallin. The quinone oxidoreductase activity of purified his-tagged recombinant mouse zeta-crystallin was comparable to that of purified native guinea pig lens zeta-crystallin, and to that previously reported for recombinant guinea pig zeta-crystallin. The method permits production of substantial amounts of recombinant zeta-crystallin for conducting studies on the biological role of this interesting protein, which exists in such high concentration in the lenses of certain species. PMID- 19679190 TI - Evidence that juvenile myoclonic epilepsy is a disorder of frontotemporal corticothalamic networks. AB - The purpose of this study is to determine regions of cerebral cortex activated during the onset and propagation of dense array electroencephalographic (dEEG) epileptiform discharges in patients with juvenile myoclonic epilepsy (JME), through the use of 256 channel, dense array scalp EEG recordings. Ten patients (16-58 years old) with the clinical diagnosis of JME comprised the study group. In all cases the MRI and neurological exams were normal, while standard EEG recordings documented typical "generalized" 4-6 Hz epileptiform patterns. Outpatient dEEG recordings captured epileptiform discharges in each patient. Localization of onset and spread of discharges in relation to a standard MRI model was accomplished by applying dipole fits and a distributed linear inverse method of cortically constrained source analysis. All patients showed epileptiform discharges that localized to sources that included orbitofrontal/medial frontopolar cortex, while basal-medial temporal lobe sources were observed in 5/10 subjects. In many ways similar to discharges of typical absence, epileptiform patterns in JME are usually irregular and frequently include temporal lobe structures as the dominant contributors to the discharges. We find that epileptiform discharges in patients with JME are not "generalized" in the sense of bilaterally synchronous diffuse onset. Rather, discharges have both localized onsets and a restricted cortical network during propagation that includes regions of frontal and temporal cortex. PMID- 19679189 TI - Progressive logopenic/phonological aphasia: erosion of the language network. AB - The primary progressive aphasias (PPA) are paradigmatic disorders of language network breakdown associated with focal degeneration of the left cerebral hemisphere. Here we addressed brain correlates of PPA in a detailed neuroanatomical analysis of the third canonical syndrome of PPA, logopenic/phonological aphasia (LPA), in relation to the more widely studied clinico-anatomical syndromes of semantic dementia (SD) and progressive nonfluent aphasia (PNFA). 32 PPA patients (9 SD, 14 PNFA, 9 LPA) and 18 cognitively normal controls had volumetric brain MRI with regional volumetry, cortical thickness, grey and white matter voxel-based morphometry analyses. Five of nine patients with LPA had cerebrospinal fluid biomarkers consistent with Alzheimer (AD) pathology (AD-PPA) and 2/9 patients had progranulin (GRN) mutations (GRN-PPA). The LPA group had tissue loss in a widespread left hemisphere network. Compared with PNFA and SD, the LPA group had more extensive involvement of grey matter in posterior temporal and parietal cortices and long association white matter tracts. Overlapping but distinct networks were involved in the AD-PPA and GRN-PPA subgroups, with more anterior temporal lobe involvement in GRN-PPA. The importance of these findings is threefold: firstly, the clinico-anatomical entity of LPA has a profile of brain damage that is complementary to the network-based disorders of SD and PNFA; secondly, the core phonological processing deficit in LPA is likely to arise from temporo-parietal junction damage but disease spread occurs through the dorsal language network (and in GRN-PPA, also the ventral language network); and finally, GRN mutations provide a specific molecular substrate for language network dysfunction. PMID- 19679191 TI - Longitudinal changes in grey and white matter during adolescence. AB - Brain development continues actively during adolescence. Previous MRI studies have shown complex patterns of apparent loss of grey matter (GM) volume and increases in white matter (WM) volume and fractional anisotropy (FA), an index of WM microstructure. In this longitudinal study (mean follow-up=2.5+/-0.5 years) of 24 adolescents, we used a voxel-based morphometry (VBM)-style analysis with conventional T1-weighted images to test for age-related changes in GM and WM volumes. We also performed tract-based spatial statistics (TBSS) analysis of diffusion tensor imaging (DTI) data to test for age-related WM changes across the whole brain. Probabilistic tractography was used to carry out quantitative comparisons across subjects in measures of WM microstructure in two fiber tracts important for supporting speech and motor functions (arcuate fasciculus [AF] and corticospinal tract [CST]). The whole-brain analyses identified age-related increases in WM volume and FA bilaterally in many fiber tracts, including AF and many parts of the CST. FA changes were mainly driven by increases in parallel diffusivity, probably reflecting increases in the diameter of the axons forming the fiber tracts. FA values of both left and right AF (but not of the CST) were significantly higher at the end of the follow-up than at baseline. Over the same period, widespread reductions in the cortical GM volume were found. These findings provide imaging-based anatomical data suggesting that brain maturation in adolescence is associated with structural changes enhancing long-distance connectivities in different WM tracts, specifically in the AF and CST, at the same time that cortical GM exhibits synaptic "pruning". PMID- 19679192 TI - CNS activation and regional connectivity during pantomime observation: no engagement of the mirror neuron system for deaf signers. AB - Deaf signers have extensive experience using their hands to communicate. Using fMRI, we examined the neural systems engaged during the perception of manual communication in 14 deaf signers and 14 hearing non-signers. Participants passively viewed blocked video clips of pantomimes (e.g., peeling an imaginary banana) and action verbs in American Sign Language (ASL) that were rated as meaningless by non-signers (e.g., TO-DANCE). In contrast to visual fixation, pantomimes strongly activated fronto-parietal regions (the mirror neuron system, MNS) in hearing non-signers, but only bilateral middle temporal regions in deaf signers. When contrasted with ASL verbs, pantomimes selectively engaged inferior and superior parietal regions in hearing non-signers, but right superior temporal cortex in deaf signers. The perception of ASL verbs recruited similar regions as pantomimes for deaf signers, with some evidence of greater involvement of left inferior frontal gyrus for ASL verbs. Functional connectivity analyses with left hemisphere seed voxels (ventral premotor, inferior parietal lobule, fusiform gyrus) revealed robust connectivity with the MNS for the hearing non-signers. Deaf signers exhibited functional connectivity with the right hemisphere that was not observed for the hearing group for the fusiform gyrus seed voxel. We suggest that life-long experience with manual communication, and/or auditory deprivation, may alter regional connectivity and brain activation when viewing pantomimes. We conclude that the lack of activation within the MNS for deaf signers does not support an account of human communication that depends upon automatic sensorimotor resonance between perception and action. PMID- 19679193 TI - Phylogenetic relationships of Ansonia from Southeast Asia inferred from mitochondrial DNA sequences: systematic and biogeographic implications (Anura: Bufonidae). AB - We investigated the phylogenetic relationships and estimated the history of species diversification and biogeography in the bufonid genus Ansonia from Southeast Asia, a unique organism with tadpoles adapted to life in strong currents chiefly in montane regions and also in lowland rainforests. We estimated phylogenetic relationships among 32 named and unnamed taxa using 2461bp sequences of the mitochondrial 12S rRNA, tRNA(val), and 16S rRNA genes with equally weighted parsimony, maximum likelihood, and Bayesian methods of inference. Monophyletic clades of Southeast Asian members of the genus Ansonia are well supported, allowing for the interpretation of general biogeographic conclusions. The genus is divided into two major clades. One of these contains two reciprocally monophyletic subclades, one from the Malay Peninsula and Thailand and the other from Borneo. The other major clade primarily consists of Bornean taxa but also includes a monophyletic group of two Philippine species and a single peninsular Malaysian species. We estimated absolute divergence times using Bayesian methods with external calibration points to reconstruct the relative timing of faunal exchange between the major landmasses of Southeast Asia. PMID- 19679194 TI - Cpc1 mediates cross-pathway control independently of Mbf1 in Fusarium fujikuroi. AB - The deletion of glnA, encoding the glutamine synthetase (GS), had led to the down regulation of genes involved in secondary metabolism and up-regulation of cpc1, the cross-pathway control transcription factor. In the present study, a Deltacpc1 mutant was created and used for transcriptional profiling by macroarray analysis. Most of the Cpc1 target genes were amino acid biosynthesis genes besides a homologue of the multi-protein bridging factor MBF1 that binds to the yeast Cpc1 homologue GCN4. We show that Deltambf1 mutants exhibit no Cpc1-related phenotype and that both proteins do not interact with each other in Fusarium fujikuroi. Moreover, results presented here suggest that Cpc1 is not responsible for the GS dependent down-regulation of secondary metabolism and that its role is focused on the activation of amino acid biosynthesis in response to the amino acid status of the cell. Surprisingly, cross-pathway control is repressed by nitrogen limitation in an AreA-dependent manner. PMID- 19679195 TI - The resveratrol analogue 4,4'-dihydroxy-trans-stilbene inhibits cell proliferation with higher efficiency but different mechanism from resveratrol. AB - Resveratrol (3,4',5-trihydroxy-trans-stilbene) is a natural phytoalexin found in grapes and wine, which shows antiproliferative activity. We previously found that 4-hydroxy group in the trans conformation was absolutely required for the inhibition of cell proliferation. In the present work we have synthesized the resveratrol analogue 4,4'-dihydroxy-trans-stilbene, which contains two OH in 4' and 4 positions, with the aim of developing a compound with an antiproliferative potential higher than that of resveratrol, on the basis of the correlation between structure and activity previously observed. In comparison with resveratrol, 4,4'-dihydroxy-trans-stilbene inhibited cell clonogenic efficiency of fibroblasts nine times more although with a different mechanism. First, 4,4' dihydroxy-trans-stilbene induced predominantly an accumulation of cells in G1 phase, whereas resveratrol perturbed the G1/S phase transition. Second, although both compounds were able to inhibit DNA polymerase (pol) delta in an in vitro assay, 4, 4'-dihydroxy-trans-stilbene did not affect pol alpha activity. Finally, 4,4'-dihydroxy-trans-stilbene increased p21(CDKN1A) and p53 protein levels, whereas resveratrol led to phosphorylation of the S-phase checkpoint protein Chk1. Taken together, our results demonstrated for the first time that the two hydroxyl groups on 4- and 4'- positions of the stilbenic backbone enhance the antiproliferative effect and introduce additional targets in the mechanism of action of resveratrol. In conclusion, 4,4'-dihydroxy-trans-stilbene has potent antiproliferative activities that differ from the effect of resveratrol shown in this system, suggesting that it warrants further development as a potential chemopreventive or therapeutic agent. PMID- 19679196 TI - Proton transfer function of carbonic anhydrase: Insights from QM/MM simulations. AB - Recent QM/MM analyses of proton transfer function of human carbonic anhydrase II (CAII) are briefly reviewed. The topics include a preliminary analysis of nuclear quadrupole coupling constant calculations for the zinc ion and more detailed analyses of microscopic pK(a) of the zinc-bound water and free energy profile for the proton transfer. From a methodological perspective, our results emphasize that performing sufficient sampling is essential to the calculation of all these quantities, which reflects the well solvated nature of CAII active site. From a mechanistic perspective, our analyses highlight the importance of electrostatics in shaping the energetics and kinetics of proton transfer in CAII for its function. We argue that once the pK(a) for the zinc-bound water is modulated to be in the proper range (approximately 7.0), proton transfer through a relatively well solvated cavity towards/from the protein surface (His64) does not require any major acceleration. Therefore, although structural details like the length of the water wire between the donor and acceptor groups still may make a non negligible contribution, our computational results and the framework of analysis suggest that the significance of such "fine-tuning" is likely secondary to the modulation of pK(a) of the zinc-bound water. We encourage further experimental analysis with mutation of (charged) residues not in the immediate neighborhood of the zinc ion to quantitatively test this electrostatics based framework; in particular, Phi analysis based on these mutations may shed further light into the relative importance of the classical Grotthus mechanism and the "proton hole" pathway that we have proposed recently for CAII. PMID- 19679197 TI - Expression and molecular dynamics studies on effect of amino acid substitutions at Arg344 in human cathepsin A on the protein local conformation. AB - Human lysosomal protective protein/cathepsin A (CathA) is a multifunctional protein that exhibits not only protective functions as to lysosomal glycosidases, i.e., neuraminidase 1 (NEU1) and beta-galactosidase (GLB), but also its own serine carboxypeptidase activity, and exhibits conserved structural similarity to yeast and wheat homologs (CPY and CPW). Our previous study revealed that the R344 (Arg344) residue in CathA could contribute to the binding and recognition of the serine peptidase inhibitor chymostatin. We examined here the effects of substitution of R344 with other amino acids, including A, D, E, G, I, K, M, N, P, Q, S, and V, denoted as R344X, including the wild-type CathA, on expression of CathA activity and intracellular processing. Among the mutant gene products, the 54-kDa precursor/zymogen with the R344D substitution was not processed to the 32/20-kDa mature form with CathA activity in a fibroblastic cell line derived from a galactosialidosis patient. Molecular dynamics (MD) simulations on the total twelve R344X mutants and the wild-type revealed that only R344D takes on a significantly different conformation of S293-D295 in the excision peptide (M285 R298) compared to the other R344X mutants; the side chains of S293 and D295 in R344D are exposed on the molecular surface, although those in the other twelve R344X mutants are buried inside the protein. The results of the current work strongly suggest that the distinct conformational change of the S293-D295 region in the R344D protein causes the processing defect of the 54-kDa precursor of the R344D mutant gene product in cultured cells. PMID- 19679198 TI - Sequestration of carbon dioxide by the hydrophobic pocket of the carbonic anhydrases. AB - The interaction between carbon dioxide (CO(2)) and the alpha-class carbonic anhydrase, human CA 2 (HCA2) exists for only a short period due to the rapid catalytic turnover by this enzyme. The fleeting nature of this interaction has led to difficulties in its direct analysis, with previous studies placing the CO(2) in the hydrophobic pocket of HCA2's active site. A more precise location was determined via the crystal structure of CO(2) trapped in both wild-type (holo) and zinc-free (apo) HCA2. This provided a detailed description of the means by which CO(2) is held and orientated for optimal catalysis. This information can be extended to the beta and gamma class enzymes to help elucidate the binding mode of CO(2) in these enzymes. PMID- 19679199 TI - Proton transfer in catalysis and the role of proton shuttles in carbonic anhydrase. AB - The undisputed role of His64 in proton transfer during catalysis by carbonic anhydrases in the alpha class has raised questions concerning the details of its mechanism. The highly conserved residues Tyr7, Asn62, and Asn67 in the active site cavity function to fine tune the properties of proton transfer by human carbonic anhydrase II (HCA II). For example, hydrophobic residues at these positions favor an inward orientation of His64 and a low pK(a) for its imidazole side chain. It appears that the predominant manner in which this fine tuning is achieved in rate constants for proton transfer is through the difference in pK(a) between His64 and the zinc-bound solvent molecule. Other properties of the active site cavity, such as inward and outward conformers of His64, appear associated with the change in DeltapK(a); however, there is no strong evidence to date that the inward and outward orientations of His64 are in themselves requirements for facile proton transfer in carbonic anhydrase. PMID- 19679200 TI - Carbonic anhydrase IX: Biochemical and crystallographic characterization of a novel antitumor target. AB - Isoform IX of the zinc enzyme carbonic anhydrase (CA, EC 4.2.1.1), CA IX, is a transmembrane protein involved in solid tumor acidification through the HIF 1alpha activation cascade. CA IX has a very high catalytic activity for the hydration of carbon dioxide to bicarbonate and protons, even at acidic pH values (of around 6.5), typical of solid, hypoxic tumors, which are largely unresponsive to classical chemo- and radiotherapy. Thus, CA IX is used as a marker of tumor hypoxia and as a prognostic factor for many human cancers. CA IX is involved in tumorigenesis through many pathways, such as pH regulation and cell adhesion control. The X-ray structure of the catalytic domain of CA IX has been recently reported, being shown that CA IX has a typical alpha-CA fold. However, the CA IX structure differs significantly from the other CA isozymes when the protein quaternary structure is considered. Thus, two catalytic domains of CA IX associate to form a dimer, which is stabilized by the formation of an intermolecular disulfide bond. The active site clefts and the proteoglycan (PG) domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. As all mammalian CAs, CA IX is inhibited by several main classes of inhibitors, such as the inorganic anions, the sulfonamides and their bioisosteres (sulfamates, sulfamides, etc.), the phenols, and the coumarins. The mechanism of inhibition with all these classes of compounds is understood at the molecular level, but the sulfonamides and their congeners have important applications. It has been recently shown that both in vitro, in cell cultures, as well as in animals with transplanted tumors, CA IX inhibition with sulfonamides lead to a return of the extracellular pH to more normal values, which leads to a delay in tumor growth. As a consequence, CA IX represents a promising antitumor target for the development of anticancer agents with an alternative mechanism of action. PMID- 19679201 TI - Structure and catalytic mechanism of the beta-carbonic anhydrases. AB - The beta-carbonic anhydrases (beta-CAs) are a diverse but structurally related group of zinc-metalloenzymes found in eubacteria, plant chloroplasts, red and green algae, and in the Archaea. The enzyme catalyzes the rapid interconversion of CO(2) and H(2)O to HCO(3)(-) and H(+), and is believed to be associated with metabolic enzymes that consume or produce CO(2) or HCO(3)(-). For many organisms, beta-CA is essential for growth at atmospheric concentrations of CO(2). Of the five evolutionarily distinct classes of carbonic anhydrase, beta-CA is the only one known to exhibit allosterism. Here we review the structure and catalytic mechanism of beta-CA, including the structural basis for allosteric regulation. PMID- 19679202 TI - Synthesis and structural characterization of strontium- and magnesium-co substituted beta-tricalcium phosphate. AB - The synthesis of five different Sr(2+)- and Mg(2+)-co-substituted beta-tricalcium phosphate (beta-TCP) has been obtained by heating the calcium-deficient apatites above 800 degrees C. With the investigated concentrations of Sr(2+) and Mg(2+) from the present study, no additional phases other than beta-TCP have been detected. The synthesized powders have been characterized by X-ray diffraction, Fourier transform infrared spectrometry, elemental analysis and Rietveld refinement studies. The co-substitution of Sr(2+) and Mg(2+) in the beta-TCP has resulted in the formation of crystalline beta-TCP at hexagonal setting (space group R3c). The reduction of lattice a- and c-axis parameters with the combined substitution of Sr(2+) and Mg(2+) in the beta-TCP has been found evident from the present results. Sr(2+) has been found occupying the Ca(1,2,3,4) sites and Mg(2+) was found at the sixfold coordinated Ca(5) site of beta-TCP structure. PMID- 19679203 TI - Interpreting humanity's genes. AB - Genetic medicine is said to be entering another era. Recent technological developments such as high resolution array techniques and next-generation sequencing have dramatically increased the power of genetic testing. However, the function of the majority of genes remains unknown. The complex interactions underpinning gene expression in humans can be studied only in part by laboratory and animal studies, and will require studies in humans. Consequently, observational studies which systematically record human phenotype data are urgently needed to interpret molecular genetic variation. PMID- 19679204 TI - Biocatalysis and agricultural biotechnology. Preface. PMID- 19679205 TI - Patterns of FGF-23, DMP1, and MEPE expression in patients with chronic kidney disease. AB - Fibroblast growth factor 23 (FGF-23), dentin matrix protein 1 (DMP1), and matrix extracellular phosphoglycoprotein (MEPE) are skeletal proteins involved in the regulation of phosphate homeostasis and bone metabolism. Circulating FGF-23 levels are increased in patients with chronic kidney disease (CKD); however, FGF 23 skeletal expression and its regulation by DMP1 and MEPE have yet to be evaluated. Thus, expression of these three proteins was characterized by immunohistochemistry in 32 pediatric and young adult patients with CKD stages 2 5. When compared to normal controls, bone FGF-23 and DMP1 expression were increased in all stages of CKD; significant differences in bone FGF-23 and DMP1 expression were not detected between pre-dialysis CKD and dialysis patients. Bone MEPE expression in CKD did not differ from controls. FGF-23 was expressed in osteocyte cell bodies located at the trabecular periphery. DMP1 was widely expressed in osteocyte cell bodies and dendrites throughout bone. MEPE was also expressed throughout bone, but only in osteocyte cell bodies. Bone FGF-23 expression correlated directly with plasma levels of the protein (r=0.43, p<0.01) and with bone DMP1 expression (r=0.54, p<0.01) and expression of both proteins were inversely related to osteoid accumulation. Bone MEPE expression was inversely related to bone volume. In conclusion, skeletal FGF-23 and DMP1 expression are increased in CKD and are related to skeletal mineralization. The patterns of expression of FGF-23, MEPE, and DMP1 differ markedly in trabecular bone, suggesting that individual osteocytes may have specialized functions. Increases in bone FGF-23 and DMP1 expression suggest that osteocyte function is altered early in the course of CKD. PMID- 19679206 TI - Increased calcium content and inhomogeneity of mineralization render bone toughness in osteoporosis: mineralization, morphology and biomechanics of human single trabeculae. AB - The differentiation and degree of the effects of mineral content and/or morphology on bone quality remain, to a large extent, unanswered due to several microarchitectural particularities in spatial measuring fields (e.g., force transfer, trajectories, microcalli). Therefore, as the smallest basic component of cancellous bone, we focused on single trabeculae to investigate the effects of mineralization and structure, both independently and in superposition. Transiliac Bordier bone cores and T12 vertebrae were obtained from 20 females at autopsy for specimen preparation, enabling radiographical analyses, histomorphometry, Bone Mineral Density Distribution (BMDD) analyses, and trabecular singularization to be performed. Evaluated contact X-rays and histomorphometric limits from cases with osteoporotic vertebral fractures generated two subdivisions, osteoporotic (n=12, O 78 years) and non-osteoporotic (n=8, O 49 years) cases, based on fracture appearance and bone volume (BV/TV). Measurements of trabecular number (Tb.N.), trabecular separation (Tb.Sp.), trabecular thickness (Tb.Th.), trabecular bone pattern factor (TBPf) and eroded surface (ES/BS) were carried out to provide detailed structural properties of the investigated groups. The mechanical properties of 400 rod-like single vertebral trabeculae, assessed by three-point bending, were matched with mineral properties as quantified by BMDD analyses of cross-sectioned rod-like and plate-like trabeculae, both in superposition and independently. Non-osteoporotic iliac crests and vertebrae displayed linear dependency on structure parameters, whereas osteoporotic compartments proved to be non-correlated with bone structure. Independent of trabecular thickness, osteoporotic rod-like trabeculae showed decreases in Young's modulus, fracture load, yield strength, ultimate stress, work to failure and bending stiffness, along with significantly increased mean calcium content and calcium width. Non-osteoporotic trabeculae showed biomechanically beneficial properties due to a homogeneous mineralization configuration, whereas osteoporotic trabeculae predominantly demonstrated various mineralized bone packets, eroded surfaces, highly mineralized cement lines and microcracks. The Young's moduli of single trabeculae exhibited significantly negative linear correlations with trabecular thickness. Because of increased, but inhomogeneously distributed, calcium content, osteoporotic trabeculae may be subject to shear stresses that render bone fragile beyond structure impairment due to cracks and lacunae. PMID- 19679207 TI - Long-term treatment of postmenopausal osteoporosis with strontium ranelate: results at 8 years. AB - OBJECTIVES: Strontium ranelate 2 g/day has proven efficacy against vertebral and nonvertebral fracture over 5 years in postmenopausal osteoporosis, though many women require longer-term treatment. This article describes the efficacy, safety, and tolerability of this agent over 8 years. METHODS: Postmenopausal osteoporotic women having participated in the 5-year efficacy trials SOTI and TROPOS were invited to enter a 3-year open-label extension study. The results presented here focus on patients who received strontium ranelate for 8 years. RESULTS: At the extension baseline, the population treated for 8 years (n=879; 79.1+/-5.6 years) had femoral neck T-score of -2.61+/-0.71. The cumulative incidences of new vertebral and nonvertebral fractures (13.7% and 12.0%, respectively) over years 6 to 8 were non-statistically different from the cumulative incidences in the first 3 years of the original studies (11.5% and 9.6%). Lumbar spine, femoral neck, and total hip bone mineral density (BMD) increased throughout the 8-year period. Annual relative change in BMD was significant at every visit, except the 8-year visit for femoral neck and total hip BMD. Strontium ranelate was safe and well tolerated over 8 years. CONCLUSIONS: Long-term treatment with strontium ranelate 2 g/day in postmenopausal osteoporotic women leads to continued increases in BMD at all sites. The data also provide some evidence for a sustained antifracture efficacy. PMID- 19679208 TI - Hydroxyapatite particles maintain peri-implant bone mantle during osseointegration in osteoporotic bone. AB - In osteoporotic bones, resorption exceeds formation during the remodelling phase of bone turnover. As a consequence, decreased bone volume and bone contact result in the peri-implant region. This may subsequently lead to loss of fixation. In this study we investigated whether the presence of nonresorbable, osteoconductive hydroxyapatite (HA) particles could help maintain a denser and more functional peri-implant bone structure. Titanium screws were implanted into the proximal tibial metaphysis of four months old, ovariectomized Wistar rats (n=60). In the right tibia, the drill hole was first filled with HA particles, while the left tibia served as a control without HA particles. Histological analysis demonstrated that during the remodelling phase the amount of newly formed bone was significantly higher on the HA over the control side. Micro-CT analysis corroborated the significant changes over time as well as differences in peri implant bone volume density between treatment and control group. Mechanical tests demonstrated that the pull-out force was greater with HA particles. These results indicate that HA particles are able to induce and maintain for a longer time a denser peri-implant bone mantle in osteoporotic bone, which may have important implications in the prevention of implant migration and cut-outs. PMID- 19679209 TI - Effects of polymorphisms of the sex hormone-binding globulin (SHBG) gene on free estradiol and bone mineral density. AB - BACKGROUND: Polymorphisms of the sex hormone-binding globulin (SHBG) gene are associated with differences in SHBG levels, influencing the risk for breast cancer and polycystic ovarian syndrome, but no association has been reported for osteoporosis in postmenopausal women. OBJECTIVE: To determine the effect of G to A substitution in the 5'UTR (rs1799941) and the Asp356Asn (rs6259) polymorphisms of the SHBG gene on bone mineral density (BMD). METHODS: This is a cross sectional study in a university-based research center from May, 2002 to December, 2007. A total of two hundred and thirteen healthy postmenopausal Caucasian women > or = 1 year from last menstrual period participated to this study. Serum estradiol by ultrasensitive radioimmnunoassay, serum sex hormone-binding globulin by immunoradiometric assay, and urinary NTx by enzyme-linked immunoassay were measured. BMD measurements were performed by dual energy X-ray absorptiometry and genotyping by Pyrosequencing. RESULTS: There were no significant differences in SHBG levels associated with either rs1799941 or rs6259. Using a p value of <0.00625 for significance, we found that subjects with the A allele (GA+AA) for the rs1799941, had a trend for lower free estradiol index (FEI) compared to the GG genotype (p=0.04). They also had significantly lower BMD at the intertrochanter (p=0.003) and trend for lower BMD at the total hip (p=0.02). There was no significant difference in FEI levels between the genotypes for the rs6259 polymorphism, but women with the Asn allele (Asp/Asn+Asn/Asn), had significantly lower BMD in the total femur (p=0.004) and intertrochanter (0.002) compared to those with the Asp/Asp genotype. CONCLUSIONS: Our data suggest that polymorphisms of the SHBG gene are associated with significant differences in BMD at the proximal femur sites. Thus, genetic variations in the SHBG gene may influence BMD at the hip in postmenopausal women. PMID- 19679210 TI - Sex-specific compromised bone healing in female rats might be associated with a decrease in mesenchymal stem cell quantity. AB - INTRODUCTION: The clinically known importance of patient sex as a major risk factor for compromised bone healing is poorly reflected in animal models. Consequently, the underlying cellular mechanisms remain elusive. Because mesenchymal stem cells (MSCs) are postulated to regulate tissue regeneration and give rise to essential differentiated cell types, they may contribute to sex specific differences in bone healing outcomes. METHODS: We investigated sex specific variations in bone healing and associated differences in MSC populations. A 1.5 mm osteotomy gap in the femora of 8 male and 8 female 12-month old Sprague-Dawley rats was stabilized by an external fixator. Healing was analyzed in terms of biomechanical testing, bridging and callus size over time (radiography at 2, 4, and 6 weeks after surgery), and callus volume and geometry by microCT at final follow-up. MSCs were obtained from bone marrow samples of an age-matched group of 12 animals (6 per gender) and analyzed for numbers of colony forming units (CFUs) and their capacity to differentiate and proliferate. The proportion of senescent cells was determined by beta-galactosidase staining. RESULTS: Sex-specific differences were indicated by a compromised mechanical competence of the callus in females compared with males (maximum torque at failure, p=0.028). Throughout the follow-up, the cross-sectional area of callus relative to bone was reduced in females (p< or =0.01), and the bridging of callus was delayed (p(2weeks)=0.041). microCT revealed a reduced callus size (p=0.003), mineralization (p=0.003) and polar moment of inertia (p=0.003) in female animals. The female bone marrow contained significantly fewer MSCs, represented by low CFU numbers in both femora and tibiae (p(femur)=0.017, p(tibia)=0.010). Functional characteristics of male and female MSCs were similar. CONCLUSION: Biomechanically compromised and radiographically delayed bone formation were distinctive in female rats. These differences were concomitant with a reduced number of MSCs, which may be causative for the suboptimal bone healing. PMID- 19679211 TI - Rapid and robust response of biochemical markers of bone formation to teriparatide therapy. AB - Teriparatide, a parathyroid hormone analogue, is a potent anabolic treatment for postmenopausal osteoporosis. Studies have shown that teriparatide induces large increases in biochemical markers of bone formation after 1 month of therapy followed by a delayed increase in bone resorption markers. The aims of this study were to (1) describe changes in bone turnover markers during 28 days of treatment with teriparatide; (2) identify the earliest time point by which most subjects showed a biochemical response to teriparatide; (3) identify potential biomarkers of positive bone response; (4) describe changes in bone turnover markers 4 weeks after stopping teriparatide. We recruited 15 osteopenic postmenopausal women, ages 55-69 (mean 62) years. All received 20 microg teriparatide subcutaneously for 28 days. Serum levels of the bone formation markers type I collagen N terminal propeptide (PINP), type I collagen C-terminal propeptide (PICP), osteocalcin (OC), bone alkaline phosphatase (bone ALP), and the bone resorption markers crosslinked C-telopeptide of type I collagen (Sbeta-CTX), crosslinked N telopeptide of type I collagen (S-NTX) and tartrate-resistant acid phosphatase type 5b (TRACP5b) were measured on 11 occasions: three times before dosing (baseline) and on days 3, 7, 10, 14, 19, 24 and 28 and at day 56 (i.e., 28 days after stopping teriparatide ). During the first 2 days of teriparatide treatment, PINP levels increased rapidly, by 8.2% (90% confidence interval (CI) 6.9%, 9.5%) and continued to increase until the end of treatment to 110.8%. PICP and OC showed a similar, but less pronounced, pattern. All three markers increased by at least 75% at day 28. A small, transient decrease in bone resorption markers occurred over the same period. Following cessation of treatment, concentrations of bone formation markers decreased to within 20% of baseline values by day 56. In conclusion, the bone formation markers PINP, PICP and OC show a rapid and robust increase in response to teriparatide, which is noticeable during the first week of therapy. PINP is the most responsive marker. These findings have important implications for monitoring patients treated with teriparatide and may also inform the design of studies of new anabolic agents for osteoporosis. PMID- 19679212 TI - The role of Akt1 in terminal stages of endochondral bone formation: angiogenesis and ossification. AB - Longitudinal bone growth is the result of endochondral bone formation which takes place in the growth plate. The rate of chondrocyte proliferation and hypertrophy, vascular invasion with the formation of primary ossification centers and cartilage replacement by bone tissue are all important processes required for normal growth. We have shown a role for the PI3K signaling pathway in chondrocyte hypertrophy and bone growth in tibia explant cultures. In this current study, we aimed to investigate the role of Akt1, an important target of PI3K, in endochondral ossification. Akt1 KO mice showed reduced size compared to their littermates throughout life, but the largest difference in body size was observed around 1 week of age. Focusing on this specific developmental stage, we discovered delayed secondary ossification in the long bones of Akt1 KO mice. A delay in formation of a structure resembling a secondary ossification center was also seen in tibia organ cultures treated with the PI3K inhibitor LY294002. The expression of matrix metalloproteinase-14 (MMP-14), the main protease responsible for development of secondary ossification centers, was decreased in the epiphysis of Akt1 KO mice, possibly explaining the delay in secondary ossification centers seen in the Akt1 KO mice. Bone mineral density (BMD) and bone mineral content (BMC) measured in the proximal tibia of 1-year-old mice were decreased in Akt1 KO mice, suggesting that the original delay in ossification might affect bone quality in older animals. PMID- 19679214 TI - Enhanced humoral and Type 1 cellular immune responses with Fluzone adjuvanted with a synthetic TLR4 agonist formulated in an emulsion. AB - Impairments in anti-influenza T helper 1 (Th1) responses are associated with greater risk of influenza-related mortality in the elderly. Addition of adjuvants to existing influenza vaccines could improve immune responses in the elderly. In this study, the activity of three adjuvants, an oil-in-water emulsion and a synthetic lipid A adjuvant formulated with or without the emulsion, is compared. Our results show that Fluzone combined with lipid A plus an emulsion effectively leads to greater vaccine-specific IgG2a and IgG titers, enhances hemagglutination inhibition titers and induces Type 1 cytokine responses (IFN-gamma and IL-2) to each of the Fluzone components. PMID- 19679213 TI - Notch signaling and the bone marrow hematopoietic stem cell niche. AB - Recently there has been increased interest in the regulatory interactions between osteoblasts and cells in the surrounding bone marrow microenvironment. The proximity of hematopoietic stem cells (HSCs) with osteoblastic cells first suggested regulatory interactions, and recent data have highlighted the role of osteoblastic cells in providing a HSC niche. Reports have indicated that direct contact is necessary to mediate the osteoblastic effects and that these effects could be mediated through Notch activation. Notch signaling is important throughout development and also appears to play a critical role in cellular maturation and differentiation of osteoblastic cells and hematopoietic cells as disregulation can lead to bone loss and leukemias, respectively. In this review we discuss the current understanding of Notch signaling and how it functions in hematopoiesis, osteoblastic cells, and the interactions between HSC and their osteoblastic niche. PMID- 19679215 TI - Viscerotropic disease following yellow fever vaccination in Peru. AB - Five suspected cases of yellow fever vaccine-associated viscerotropic disease (YEL-AVD) clustered in space and time following a vaccination campaign in Ica, Peru in 2007. All five people received the same lot of 17DD live attenuated yellow fever vaccine before their illness; four of the five died of confirmed YEL AVD. The surviving case was classified as probable YEL-AVD. Intensive investigation yielded no abnormalities of the implicated vaccine lot and no common risk factors. This is the first described space-time cluster of yellow fever viscerotropic disease involving more than two cases. Mass yellow fever vaccination should be avoided in areas that present extremely low risk of yellow fever. PMID- 19679216 TI - Safety and efficacy of human rotavirus vaccine during the first 2 years of life in Asian infants: randomised, double-blind, controlled study. AB - This study evaluates the safety and efficacy against severe rotavirus gastroenteritis of the oral live attenuated human rotavirus vaccine RIX4414 (Rotarix) during the first 2 years of life in Asian infants from high-income countries. Healthy infants were enrolled to receive 2 doses of RIX4414 (N=5,359) or placebo (N=5,349). From 2 weeks post-dose 2 to 2 years of age, vaccine efficacy was 96.1% (95%CI:85.1%; 99.5%) against severe rotavirus gastroenteritis, 100% (95%CI:80.8%; 100%) against wild-type G1P[8] and 93.6% (95%CI:74.7%; 99.3%) against circulating non-G1 rotavirus types. No intussusception cases were reported within 31 days post-vaccination. RIX4414 shows a good safety profile and offers high protection during the first 2 years of life with potentially significant public health impact in this population. PMID- 19679217 TI - Reduction in hepatitis B virus seroprevalence among U.S.-born children of foreign born Asian parents -- benefit of universal infant hepatitis B vaccination. AB - We demonstrate that after implementation of recommendations for universal infant hepatitis B vaccination, HBV infection prevalence among children of foreign-born Asian parents in Georgia declined dramatically; horizontal transmission of infection within households has occurred infrequently; and the vast majority of infants and children have received the recommended hepatitis B vaccinations. These results provide evidence of the success of the hepatitis B infant vaccination program and highlight its potential impact on reducing chronic HBV infection morbidity and mortality among U.S. populations at high risk. PMID- 19679218 TI - Control of pertussis--lessons learnt from a 10-year surveillance programme in Sweden. AB - Sweden was the only country in the world without any general pertussis vaccination when acellular pertussis (aP) vaccines were introduced. Since 1996 aP vaccines are given at the ages of 3, 5 and 12 months, with a 99% coverage, and until 2007 without a later booster. The long-term effects of aP vaccines, monitored within an enhanced surveillance project, were discussed at an international workshop in Stockholm in November 2008. The unique Swedish experience demonstrates that aP vaccines are capable of achieving the primary goal of a national vaccination programme, i.e., to significantly reduce the burden of pertussis in pre-school children. Throughout the 10-year surveillance period the highest age-specific incidence was reported in unvaccinated infants or those who had received only one dose, with most hospitalisations in this age group and eight deaths among unvaccinated infants. Complementary strategies are needed to achieve further reduction in morbidity from circulation of Bordetella pertussis. PMID- 19679219 TI - Does receipt of seasonal influenza vaccine predict intention to receive novel H1N1 vaccine: evidence from a nationally representative survey of U.S. adults. AB - We analyze data on the intention of U.S. adults to receive novel H1N1 vaccine if available this fall, and studies the relationship between the intention to be vaccinated against novel H1N1 and the uptake of seasonal influenza vaccine last year. We surveyed a nationally representative sample of U.S. adults (n=2067) via the Internet between May 26th and June 8th, 2009. Our results imply a vaccination rate for novel H1N1 of 49.6%, which corresponds to roughly 115 million adult vaccinations. Moreover, novel H1N1 vaccination intentions are strongly associated with seasonal influenza vaccinations, suggesting common attitudinal barriers to both vaccines. PMID- 19679220 TI - Development of an ex vivo human skin model for intradermal vaccination: tissue viability and Langerhans cell behaviour. AB - The presence of resident Langerhans cells (LCs) in the epidermis makes the skin an attractive target for DNA vaccination. However, reliable animal models for cutaneous vaccination studies are limited. We demonstrate an ex vivo human skin model for cutaneous DNA vaccination which can potentially bridge the gap between pre-clinical in vivo animal models and clinical studies. Cutaneous transgene expression was utilised to demonstrate epidermal tissue viability in culture. LC response to the culture environment was monitored by immunohistochemistry. Full thickness and split-thickness skin remained genetically viable in culture for at least 72 h in both phosphate-buffered saline (PBS) and full organ culture medium (OCM). The epidermis of explants cultured in OCM remained morphologically intact throughout the culture duration. LCs in full-thickness skin exhibited a delayed response (reduction in cell number and increase in cell size) to the culture conditions compared with split-thickness skin, whose response was immediate. In conclusion, excised human skin can be cultured for a minimum of 72 h for analysis of gene expression and immune cell activation. However, the use of split thickness skin for vaccine formulation studies may not be appropriate because of the nature of the activation. Full-thickness skin explants are a more suitable model to assess cutaneous vaccination ex vivo. PMID- 19679223 TI - Complementary and alternative medicine (CAM) and cancer: the ugly face of alternative medicine. AB - This essay describes my thoughts on the role of CAM in cancer and was provoked by some ugly scenes when I was debating the subject at Kings College London. I conclude that when one analyzes what CAM truly has to offer compared with the best of conventional medicine, then CAM does more harm than good. However it is up to modern medical practitioners to raise their game in order to prevent practitioners of alternative medicine slipping into the gaps left unattended by our profession. PMID- 19679221 TI - Glucosamine but not ibuprofen alters cartilage turnover in osteoarthritis patients in response to physical training. AB - OBJECTIVE: To investigate changes in levels of serum cartilage oligomeric matrix protein (COMP) and urine c-telopeptide of type-2 collagen (CTX-II) as markers for cartilage turnover in patients with osteoarthritis (OA) of the knee, in response to muscle strength training in combination with treatment with glucosamine, ibuprofen or placebo. DESIGN: A 12-week double blind, placebo controlled, randomized study. METHOD: Thirty-six elderly patients with bilateral tibiofemoral knee OA determined by radiography were randomly assigned to treatment with glucosamine (n=12), ibuprofen (n=12) or placebo (n=12) during 12 weeks of strength training of both legs with focus on the quadriceps muscle. Strength tests (5 repetition maximum), blood and urine sampling were performed before and after the training period. Serum COMP and urinary CTX-II were measured by enzyme linked immunosorbent assay (ELISA). RESULTS: All three groups increased their muscle strength following 12 weeks of strength training (P<0.001). Serum COMP levels were reduced in the glucosamine-treated group after the training period (P=0.012), whereas they did not change in the two other groups. Glucosamine reduced COMP statistically significant compared to both placebo and ibuprofen; the mean reduction with glucosamine was 13% vs placebo (P=0.0378) and 17% vs ibuprofen (P=0.0122). Urinary CTX-II levels did not change significantly in any of the three experimental groups. CONCLUSION: Serum COMP decreased significantly over the 12-week training period when treatment with glucosamine was added to the training regimen. This suggests an effect by glucosamine on the response of the OA cartilage to a period of joint loading in humans with knee OA. PMID- 19679224 TI - Massively parallel sequencing: the next big thing in genetic medicine. AB - Massively parallel sequencing has reduced the cost and increased the throughput of genomic sequencing by more than three orders of magnitude, and it seems likely that costs will fall and throughput improve even more in the next few years. Clinical use of massively parallel sequencing will provide a way to identify the cause of many diseases of unknown etiology through simultaneous screening of thousands of loci for pathogenic mutations and by sequencing biological specimens for the genomic signatures of novel infectious agents. In addition to providing these entirely new diagnostic capabilities, massively parallel sequencing may also replace arrays and Sanger sequencing in clinical applications where they are currently being used. Routine clinical use of massively parallel sequencing will require higher accuracy, better ways to select genomic subsets of interest, and improvements in the functionality, speed, and ease of use of data analysis software. In addition, substantial enhancements in laboratory computer infrastructure, data storage, and data transfer capacity will be needed to handle the extremely large data sets produced. Clinicians and laboratory personnel will require training to use the sequence data effectively, and appropriate methods will need to be developed to deal with the incidental discovery of pathogenic mutations and variants of uncertain clinical significance. Massively parallel sequencing has the potential to transform the practice of medical genetics and related fields, but the vast amount of personal genomic data produced will increase the responsibility of geneticists to ensure that the information obtained is used in a medically and socially responsible manner. PMID- 19679225 TI - X chromosomal variation is associated with slow progression to AIDS in HIV-1 infected women. AB - AIDS has changed from a mostly male-specific health problem to one that predominantly affects females. Although sex differences in HIV-1 susceptibility are beyond doubt, the extent to which sex affects the onset and progression of AIDS has remained elusive. Here, we provide evidence for an influence of X chromosomal variation on the course of retroviral infection, both in HIV-1 infected patients and in the rhesus macaque model of AIDS. A two-stage, microsatellite-based GWAS of SIV-infected monkeys revealed MHC class I markers and a hitherto-unknown X chromosomal locus as being associated with a nominal score measuring progression to AIDS (Fisher's exact p < 10(-6)). The X chromosomal association was subsequently confirmed in HIV-1-infected patients with published SNP genotype data. SNP rs5968255, located at human Xq21.1 in a conserved sequence element near the RPS6KA6 and CYLC1 genes, was identified as a significant genetic determinant of disease progression in females (ANOVA p = 8.8 x 10(-5)), but not in males (p = 0.19). Heterozygous female carriers of the C allele showed significantly slower CD4 cell decline and a lower viral load at set point than TT homozygous females and than males. Inspection of HapMap revealed that the CT genotype is significantly more frequent among Asians than among Europeans or Africans. Our results suggest that, in addition to the individual innate and adaptive immunity status, sex-linked genetic variation impacts upon the rate of progression to AIDS. Elucidating the mechanisms underlying this sex specific effect will promote the development of antiretroviral therapies with high efficacy in both sexes. PMID- 19679226 TI - IDS crossing of the blood-brain barrier corrects CNS defects in MPSII mice. AB - Mucopolysaccharidosis type II (MPSII), or Hunter syndrome, arises from a deficiency in iduronate 2-sulfatase (IDS), and it is characterized by progressive somatic and neurological involvement. The MPSII mouse model reproduces the features of MPSII patients. Systemic administration of the AAV2/5CMV-hIDS vector in MPSII mouse pups results in the full correction of glycosaminoglycan (GAG) accumulation in visceral organs and in the rescue of the defects and GAG accumulation in the central nervous system (CNS). Remarkably, in treated MPSII animals, this CNS correction arises from the crossing of the blood-brain barrier by the IDS enzyme itself, not from the brain transduction. Thus, we show here that early treatment of MPSII mice with one systemic injection of AAV2/5CMV-hIDS results in prolonged and high levels of circulating IDS that can efficiently and simultaneously rescue both visceral and CNS defects for up to 18 months after therapy. PMID- 19679227 TI - Synthesis of monodeoxy and mono-O-methyl congeners of methyl beta-D mannopyranosyl-(1-->2)-beta-D-mannopyranoside for epitope mapping of anti-Candida albicans antibodies. AB - A panel of six complementary monodeoxy and mono-O-methyl congeners of methyl beta d-mannopyranosyl-(1-->2)-beta-d-mannopyranoside (1) were synthesized by stereoselective glycosylation of monodeoxy and mono-O-methyl monosaccharide acceptors with a 2-O-acetyl-glucosyl trichloroacetimidate donor, followed by a two-step oxidation-reduction sequence at C-2'. The beta-manno configurations of the final deprotected congeners 2-7 were confirmed by measurement of (1)J(C1,H1) heteronuclear and (3)J(1',2') homonuclear coupling constants. These disaccharide derivatives will be used to map the protective epitope recognized by a protective anti-Candida albicans monoclonal antibody C3.1 (IgG3) and to determine its key polar contacts with the binding site. PMID- 19679228 TI - Characterization of Acinetobacter baumannii genotypes recovered from patients with repeated colonization or infection. AB - Acinetobacter baumannii has become an important nosocomial pathogen associated with increasing prevalence of multidrug resistance. Effective strategies to prevent emerging and spreading multidrug-resistant A. baumannii have not been well developed because of limited understanding of the organism epidemiology. The aim of this study was to determine if infection/colonization with A. baumannii after the initial isolation of the organism was due to the persistence of identical strain in patients with repeated positive clinical cultures. Forty-one patients with two or more clinical isolations of A. baumannii separated by at least 30 days were identified. The genotype of the first isolate from each patient was compared with the pulsed-field gel electrophoresis types of subsequent isolates of the same patient. Persistence of a single strain was detected in 51% patients. In the remaining 49% patients, subsequent isolates were not identical to the first isolate. Patients with isolates initially identified as carbapenem (imipenem or meropenem) resistant were 5.2 times more likely to have closely related isolates obtained for all subsequent cultures than patients with initial carbapenem-intermediate or carbapenem-susceptible isolates. PMID- 19679229 TI - A rapid allele variant discrimination method for Yersinia pestis strains based on high-resolution melting curve analysis. AB - Yersinia pestis isolates were genotyped analyzing the polymorphic DNA regions named variable number tandem repeats (VNTR). Allele variants were studied by high resolution melting analysis (HRMA) of polymerase chain reaction fragments obtained for 25 VNTR loci. After comparison with previous results, 14 loci gave distinguishable normalized melting curves and allowed to correctly assign alleles. This HRMA typing technique permits to differentiate Y. pestis isolates and turned out to be robust, reproducible, and cheap. PMID- 19679230 TI - The evaluation of a user-friendly lateral flow assay for the serodiagnosis of human brucellosis in Kazakhstan. AB - Serum samples from all patients with culture-confirmed brucellosis including those with chronic disease from Kazakhstan tested positive in the serum agglutination test for titers > or = 1:25 and reacted in the Brucella immunoglobulin M/immunoglobulin G lateral flow assay (LFA) confirming the high sensitivity of these assays. The strong reactivity in the LFA observed for the majority (92.1%) of the samples from the patients with culture-confirmed brucellosis together with the user-friendliness of the assay procedure makes the LFA ideal for the confirmation of brucellosis in endemic areas in Kazakhstan. The Rose Bengal test lacked sensitivity in particular for patients with chronic brucellosis therefore limiting its value as a quick screening assay. The study emphasizes the importance of the LFA as a useful, rapid, and easy-to-perform tool in the diagnostic testing of brucellosis. PMID- 19679231 TI - Preliminary evaluation of a procedure for improved detection of Shiga toxin producing Escherichia coli in fecal specimens. AB - Culture confirmation of Shiga toxin-producing Escherichia coli (STEC) is very important for epidemiologic analysis. However, isolation of non-O157 STEC on conventional selective media such as sorbitol-MacConkey agar (SMAC) can be difficult because of heavy growth of competing bacteria and its phenotypical similarity to commensal nonpathogenic E. coli. An acid enrichment procedure was introduced in this study to facilitate detection of STEC from patients who were symptomatic. Forty-seven clinical fecal broths, which tested positive for Shiga toxin by commercial immunoassay, were processed for the isolation of STEC by both conventional and the acid enrichment methods. The acid enrichment method and conventional culture recovered STEC from 91% (43/47) and 70% (33/47) of the fecal broths, respectively. Neither method retrieved STEC in 3 specimens. Thirty-six STEC were successfully serogrouped, which included O26 (n = 11), O157 (n = 9), O103 (n = 7), O121 (n = 3), O111 (n = 2 each), O28AC, O146, O76, and O undetermined (n = 1 each). The analysis of STEC isolates by real-time PCR indicated that all 9 E. coli O157 contained stx2 gene alone or in combination with stx1. Non-O157 STEC more frequently contained stx1 only, and about one-third possessed stx2. The novel acid enrichment protocol greatly reduced the growth of competitor colonies on RTN and TCSMAC. The study demonstrated that incorporation of an acid enrichment procedure in clinical testing improved the isolation of STEC in fecal specimens. PMID- 19679233 TI - Factors affecting QuickVue Influenza A + B rapid test performance in the community setting. AB - Rapid diagnosis of influenza can facilitate timely clinical management. We evaluated the performance of the QuickVue Influenza A + B test (Quidel, San Diego, CA) in a community setting and investigated the factors affecting test sensitivity. We recruited 1008 subjects from 30 outpatient clinics in Hong Kong between February and September 2007. Each subject provided 2 pooled pairs of nose and throat swabs; 1 pair was tested by the QuickVue rapid test on site, and the other pair was sent to a laboratory for reference tests. Among 998 enrolled subjects with valid results, the rapid test had overall sensitivity of 0.68 and specificity of 0.96 compared with viral culture. Sensitivity for both influenza A and B was significantly higher for specimens with viral loads greater than 5 log(10) copies/mL. The QuickVue Influenza A + B test has similar sensitivity in point-of-care community settings to more controlled conditions. PMID- 19679234 TI - Molecular and serologic markers of acute dengue infection in naive and flavivirus vaccinated travelers. AB - A total of 520 European travelers with suspected dengue fever were examined, and acute dengue virus infection was confirmed in 127 of them. Molecular and serological tests for dengue diagnosis, according to their usefulness in the different stages of the disease, were performed. The accuracy of the IgM/IgG ratio and the IgG avidity index was confirmed during the early phase of the illness to discriminate the serologic status (primary versus secondary immune response) of patients who were either naive or previously vaccinated against other flaviviruses. Virologic markers of secondary infection were detected in 11.8% of nonvaccinated infected patients and in 92.6% of yellow fever vaccinated patients. The proper use of these simple methodologies could help in the identification of patients affected with secondary infection, who may be at a higher risk of developing a hemorrhagic disorder. PMID- 19679235 TI - Emergence of Streptococcus pneumoniae of serotype 19A in France: molecular capsular serotyping, antimicrobial susceptibilities, and epidemiology. AB - We have studied 457 Streptococcus pneumoniae isolated in 2007 from adults and children. For all isolates, both latex agglutination and molecular capsular typing were performed. Antibiotic resistance patterns were determined. S. pneumoniae 19A was the most frequently isolated serotype (34.7%) both in children and adults. It represented 12.8% of the strains isolated from invasive infections in adults and 27.0% in children and 63.6% (110/173) of strains isolated from acute otitis media. Between children and adults, no difference concerning antibiotic susceptibility was observed for penicillin, amoxicillin, cefotaxime, and erythromycin in strains isolated from invasive diseases. Comparing antibiotic susceptibilities according to the serotype, the 19A isolates appeared to be the least susceptible to penicillin (3.2%) and erythromycin (4.5%), followed by serotypes 19F and 14. We confirm the predominance of serotype 19A among S. pneumoniae responsible for invasive and noninvasive diseases either in children or adults in France. PMID- 19679236 TI - Molecular diagnosis and treatment monitoring of congenital transmission of Trypanosoma cruzi to twins of a triplet delivery. AB - Congenital transmission of Trypanosoma cruzi was diagnosed in 2 triplets born to a triamniotic bichorionic delivery. Only the sisters sharing the placenta became infected, as diagnosed by microhematocrit and/or polymerase chain reaction of 3 parasite targets. The neonates' parasitologic response to benznidazole was monitored. Molecular strategies allowed genotyping lineage IId and identical minicircle signatures in both triplets, showing they harbored the same maternal parasite populations. PMID- 19679237 TI - Lobomycosis: diagnosis and management of relapsed and multifocal lesions. AB - We report a case of lobomycosis in the left leg of a patient that had traveled to the endemic Brazilian Amazon region. After surgical resection of the lesion, there was relapse with local dissemination of the disease and the treatment was successfully performed by oral itraconazole and cryosurgery. PMID- 19679238 TI - Unrecognized cervical spinal epidural abscess associated with metastatic Klebsiella pneumoniae bacteremia and liver abscess in nondiabetic patients. AB - We describe a Klebsiella pneumoniae-related liver abscess and unrecognized epidural abscess with or without endophthalmitis in 2 nondiabetic patients, both of whom suffered neurologic complications after invasive procedure. Although rare, we should keep these types of cases in mind when making a diagnosis in patients with both liver abscess and complaint of neck pain. PMID- 19679239 TI - Real-time polymerase chain reaction detection of Cryptococcus neoformans and Cryptococcus gattii in human samples. AB - We report a TaqMan real-time polymerase chain reaction (PCR) protocol for diagnosing cryptococcosis. Specificity was tested with 33 fungal strains. The 7 clinical samples found positive by culture also tested positive by real-time PCR. No false negatives were found among the 94 clinical samples that were negative by culture for Cryptococcus spp. The sensitivity threshold was about 10 plasmid copies per assay. PMID- 19679232 TI - Comparison of results of fluconazole and voriconazole disk diffusion testing for Candida spp. with results from a central reference laboratory in the ARTEMIS DISK Global Antifungal Surveillance Program. AB - The accuracy of antifungal susceptibility testing is important for reliable resistance surveillance and for the clinical management of patients with serious infections due to Candida spp. Our primary objective was to compare the results of fluconazole and voriconazole disk diffusion testing of 3227 Candida spp. performed by 47 centers participating in the ARTEMIS program with disk diffusion and MIC results obtained by the central reference laboratory. The overall categoric agreement between participant disk diffusion test results and reference MIC results was 87% for fluconazole and 95.2% for voriconazole. Likewise good agreement was observed between participant disk diffusion test results and reference laboratory disk diffusion test results, with an agreement of 90.5%, 1% very major error (VME), and 3.4% major error (ME) for fluconazole and 94.2%, 1.1% VME, and 2.5% ME for voriconazole. The disk diffusion test was reliable for detecting those isolates of Candida spp. that were characterized as resistant to fluconazole and voriconazole by MIC testing. External quality assurance data obtained by surveillance programs such as the ARTEMIS Global Antifungal Surveillance Program ensure the generation of useful surveillance data and result in the continued improvement of antifungal susceptibility testing protocols. PMID- 19679240 TI - A diagnosis of Burkholderia pseudomallei directly in a bronchoalveolar lavage by polymerase chain reaction. AB - Melioidosis is an infectious disease caused by the Gram-negative bacterium Burkholderia pseudomallei. Interest in the molecular identification of B. pseudomallei has increased after its classification as a category B agent by the US Centers for Disease Control and Prevention. The present article reports a diagnosis of B. pseudomallei directly in a bronchoalveolar lavage by polymerase chain reaction amplification. The results obtained show that direct detection of the 16-23s spacer sequence in bronchoalveolar lavage is a quick and specific test to diagnose melioidosis. PMID- 19679242 TI - The dispersin-encoding gene (aap) is not restricted to enteroaggregative Escherichia coli. AB - The presence of the enteroaggregative Escherichia coli (EAEC) virulence genes aatA, aap, and aggR was assayed in strains of different diarrheagenic E. coli pathotypes and nonpathogenic E. coli. The dispersin-encoding gene (aap) was detected in EAEC, diffusely adherent E. coli, and nonpathogenic E. coli, demonstrating that molecular diagnostics of EAEC based on aap detection may identify non-EAEC strains. PMID- 19679241 TI - Susceptibility and bactericidal activity of 8 oral quinolones against conventional-fluoroquinolone-resistant Streptococcus pneumoniae clinical isolates. AB - We evaluated the pharmacokinetic/pharmacodynamic parameters of 8 oral quinolones (ciprofloxacin, garenoxacin [GRNX], gatifloxacin, levofloxacin, moxifloxacin, prulifloxacin, sitafloxacin, and sparfloxacin) on 11 fluoroquinolone-resistant Streptococcus pneumoniae strains, screened from 780 strains isolated from various clinical sources in Japan. GRNX showed the highest area under the blood concentration time curve/MIC ratios, which exceeded the target values for bacterial eradication against all fluoroquinolone-resistant S. pneumoniae strains. PMID- 19679243 TI - Bicuspid pulmonic valve. PMID- 19679244 TI - The CD40/CD40 ligand system: linking inflammation with atherothrombosis. AB - The role of CD40/CD40 ligand (CD40L) in atherothrombosis is now widely accepted. However, the exact mechanisms linking the CD40/CD40L system and the soluble form of CD40 ligand (sCD40L) with atherothrombosis are currently a topic of intensive research. CD40L and sCD40L belong to the tumor necrosis factor superfamily, and they are molecules with a dual prothrombotic and proinflammatory role. They are expressed in a variety of tissues such as the immune system (in both B and T cells), the vascular wall, and activated platelets. Soluble CD40L has multiple autocrine, paracrine, and endocrine actions, and it may trigger key mechanisms participating in atherothrombosis. CD40/CD40L may participate in the development of coronary atherosclerosis and the triggering of acute coronary syndromes, while sCD40L seems to have a prognostic role not only in subjects with advanced atherosclerosis but also in the general population. Although conventional cardiovascular medication such as antiplatelet therapy, statins, angiotensin converting enzyme inhibitors, and many others have been shown to reduce both sCD40L and cardiovascular risk, it is still unclear whether specific treatments targeting the CD40/CD40L system will prove to be beneficial against atherothrombosis in the near future. PMID- 19679245 TI - The efficacy and safety of prasugrel with and without a glycoprotein IIb/IIIa inhibitor in patients with acute coronary syndromes undergoing percutaneous intervention: a TRITON-TIMI 38 (Trial to Assess Improvement in Therapeutic Outcomes by Optimizing Platelet Inhibition With Prasugrel-Thrombolysis In Myocardial Infarction 38) analysis. AB - OBJECTIVES: We evaluated the efficacy and safety of prasugrel and clopidogrel in the setting of a glycoprotein (GP) IIb/IIIa inhibitor. BACKGROUND: Prasugrel reduced cardiovascular events as compared with clopidogrel in TRITON-TIMI 38 (Trial to Assess Improvement in Therapeutic Outcomes by Optimizing Platelet Inhibition with Prasugrel-Thrombolysis in Myocardial Infarction 38) but with increased bleeding. METHODS: Researchers in the TRITON-TIMI 38 randomized 13,608 subjects with acute coronary syndrome undergoing percutaneous coronary intervention to prasugrel versus clopidogrel. The use of a GP IIb/IIIa inhibitor was at the physician's discretion. For the current analysis, end points were examined at 30 days and were stratified by use of a GP IIb/IIIa inhibitor. RESULTS: A total of 7,414 subjects (54.5%) received a GP IIb/IIIa inhibitor during their index hospitalization. There was a consistent benefit of prasugrel over clopidogrel for reducing cardiovascular death, myocardial infarction, or stroke in patients who did (hazard ratio: 0.76; 95% confidence interval: 0.64 to 0.90) or did not receive a GP IIb/IIIa inhibitor (hazard ratio: 0.78; 95% confidence interval: 0.63 to 0.97, p(interaction) = 0.83). Prasugrel significantly reduced myocardial infarction, urgent revascularization, and stent thrombosis irrespective of GP IIb/IIIa inhibitor use. Although subjects treated with a GP IIb/IIIa inhibitor had greater rates of bleeding, the risk of Thrombolysis in Myocardial Infarction major or minor bleeding with prasugrel versus clopidogrel was not significantly different in patients who were or were not treated with GP IIb/IIIa inhibitor (p(interaction) = 0.19). CONCLUSIONS: Prasugrel significantly reduces the risk of cardiovascular events in patients with acute coronary syndromes after percutaneous coronary intervention regardless of whether or not a GP IIb/IIIa inhibitor is used. The use of a GP IIb/IIIa inhibitor does not accentuate the relative risk of bleeding with prasugrel as compared with clopidogrel. PMID- 19679246 TI - Percutaneous mitral repair with the MitraClip system: safety and midterm durability in the initial EVEREST (Endovascular Valve Edge-to-Edge REpair Study) cohort. AB - OBJECTIVES: We undertook a prospective multicenter single-arm study to evaluate the feasibility, safety, and efficacy of the MitraClip system (Evalve Inc., Menlo Park, California). BACKGROUND: Mitral valve repair for mitral regurgitation (MR) has been performed by the use of a surgically created double orifice. Percutaneous repair based on this surgical approach has been developed by use of the Evalve MitraClip device to secure the mitral leaflets. METHODS: Patients with 3 to 4+ MR were selected in accordance with the American Heart Association/American College of Cardiology guidelines for intervention and a core echocardiographic laboratory. RESULTS: A total of 107 patients were treated. Ten (9%) had a major adverse event, including 1 nonprocedural death. Freedom from clip embolization was 100%. Partial clip detachment occurred in 10 (9%) patients. Overall, 79 of 107 (74%) patients achieved acute procedural success, and 51 (64%) were discharged with MR of < or =1+. Thirty-two patients (30%) had mitral valve surgery during the 3.2 years after clip procedures. When repair was planned, 84% (21 of 25) were successful. Thus, surgical options were preserved. A total of 50 of 76 (66%) successfully treated patients were free from death, mitral valve surgery, or MR >2+ at 12 months (primary efficacy end point). Kaplan-Meier freedom from death was 95.9%, 94.0%, and 90.1%, and Kaplan-Meier freedom from surgery was 88.5%, 83.2%, and 76.3% at 1, 2, and 3 years, respectively. The 23 patients with functional MR had similar acute results and durability. CONCLUSIONS: Percutaneous repair with the MitraClip system can be accomplished with low rates of morbidity and mortality and with acute MR reduction to < 2+ in the majority of patients, and with sustained freedom from death, surgery, or recurrent MR in a substantial proportion (EVEREST I; NCT00209339. EVEREST II; NCT00209274). PMID- 19679247 TI - Increased wave reflection rather than central arterial stiffness is the main determinant of raised pulse pressure in women and relates to mismatch in arterial dimensions: a twin study. AB - OBJECTIVES: Our aim was to examine the relative contributions of the first systolic shoulder (P1) and augmentation pressure (DeltaP(aug)) to central pulse pressure (cPP), their relation to central arterial stiffness (pulse wave velocity [PWV]) and arterial diameters, and their respective heritability estimates. BACKGROUND: cPP is augmented above P1 by DeltaP(aug) due to pressure waves reflected from the periphery of the circulation. METHODS: Women (n = 496) from the Twins UK adult twin registry (112 monozygotic, 135 dizygotic pairs) age 21 to 81 years were studied. cPP, P1, and DeltaP(aug) were estimated using the SphygmoCor system (Atcor, West Ryde, Australia) from transformed radial waveforms. Carotid-femoral PWV was measured using the same system. Aortic and femoral artery diameters were measured by ultrasonography. Heritability was estimated using structural equation modeling. RESULTS: P1 and DeltaP(aug) accounted for 22% and 76%, respectively, of the variance in cPP. After adjustment for mean arterial pressure and heart rate, P1 strongly independently positively correlated with PWV (standardized regression coefficient, beta = 0.4, p < 0.0001), whereas DeltaP(aug) did not independently correlate with PWV but independently negatively correlated with the ratio of the diameter of the femoral to that of the abdominal aorta (beta = -0.12, p < 0.001). Estimates of heritability (h(2)) of cPP, PWV, P1, and DeltaP(aug) were 0.43, 0.34, 0.31, and 0.62, respectively, after adjustment for mean arterial pressure and heart rate. CONCLUSIONS: These results suggest that, in women, DeltaP(aug) is highly heritable, is associated with the ratio of distal to proximal arterial diameters, and, independent of PWV, is a major determinant of cPP. PMID- 19679248 TI - Impact of heart rate on central aortic pressures and hemodynamics: analysis from the CAFE (Conduit Artery Function Evaluation) study: CAFE-Heart Rate. AB - OBJECTIVES: The CAFE (Conduit Artery Function Evaluation) study showed less effective central aortic pressure lowering with atenolol-based therapy versus amlodipine-based therapy in people with hypertension. The present study examined the importance of heart rate (HR) as a determinant of this effect. BACKGROUND: Recent analyses have suggested that beta-blockers are less effective at reducing cardiovascular events than alternative blood pressure (BP)-lowering therapies. There has been much debate about the mechanism for this shortfall in benefit and specifically the role of HR lowering by beta-blockers. METHODS: Central pressures were derived from brachial pressure and radial pulse wave analysis in 2,073 patients, and 7,146 measurements were recorded and analyzed over follow-up for up to 4 years. RESULTS: There was no impact of HR on brachial systolic or pulse pressures; however, there was a highly significant inverse relationship between HR and central aortic systolic and pulse pressures (p < 0.001). This was dependent on a strong inverse relationship between HR and augmentation index, indicative of increased wave reflection at lower HRs. Multiple regression, adjusted for brachial BP, showed HR to be the major determinant of central pressures. Moreover, HR and brachial BP accounted for 92% of the variability in central systolic and pulse pressures. Consequently, drug-related differences in central aortic pressures were markedly attenuated after adjustment for HR. CONCLUSIONS: When comparing beta-blocker-based treatments with other BP-lowering strategies, HR reduction with beta-blockers is a major mechanism accounting for less effective central aortic pressure reduction per unit change in brachial pressure. PMID- 19679249 TI - The pulse of cardiology: quo vadis? PMID- 19679250 TI - Beneficial cardiovascular effects of bariatric surgical and dietary weight loss in obesity. AB - OBJECTIVES: We hypothesized that, in obese persons without comorbidities, cardiovascular responses to excess weight are reversible during weight loss by either bariatric surgery or diet. BACKGROUND: Obesity is associated with cardiac hypertrophy, diastolic dysfunction, and increased aortic stiffness, which are independent predictors of cardiovascular risk. METHODS: Thirty-seven obese (body mass index 40 +/- 8 kg/m(2)) and 20 normal-weight subjects (body mass index 21 +/ 2 kg/m(2)) without identifiable cardiac risk factors underwent cardiac magnetic resonance imaging for the assessment of the left and right ventricles and of indexes of aortic function. Thirty of the obese subjects underwent repeat imaging after 1 year of significant weight loss, achieved in 17 subjects by diet and in 13 subjects by bariatric surgery. Seven obese subjects underwent repeat imaging after 1 year of continued obesity. RESULTS: Left and right ventricular masses were significantly increased, left ventricular diastolic function impaired, and aortic distensibility reduced in the obese. Both diet and bariatric surgery led to comparable, significant decreases in left and right ventricular masses, end diastolic volume, and diastolic dysfunction, and an increase in aortic distensibility at all levels of the aorta, most pronounced distally (e.g., distal descending aorta 5.1 +/- 1.8 mm Hg(-1) x 10(-3) before weight loss and 6.8 +/- 2.5 mm Hg(-1) x 10(-3) after weight loss; p < 0.001). No improvements were observed in continued obesity. CONCLUSIONS: Irrespective of method, 1 year of weight loss leads to partial regression of cardiac hypertrophy and to reversal of both diastolic dysfunction and aortic distensibility impairment. These findings provide a potential mechanism for the reduction in mortality seen with weight loss. PMID- 19679251 TI - Obesity and cardiovascular disease: how can cardiac magnetic resonance help? PMID- 19679252 TI - Outcome of extracorporeal membrane oxygenation for early primary graft failure after pediatric heart transplantation. AB - OBJECTIVES: We sought to analyze the indications and outcome of extracorporeal membrane oxygenation (ECMO) for early primary graft failure and determine its impact on long-term graft function and rejection risk. BACKGROUND: Early post operative graft failure requiring ECMO can complicate heart transplantation. METHODS: A retrospective review of all children requiring ECMO in the early period after transplantation from 1990 to 2007 was undertaken. RESULTS: Twenty eight (9%) of 310 children who underwent transplantation for cardiomyopathy (n = 5) or congenital heart disease (n = 23) required ECMO support. The total ischemic time was significantly longer for ECMO-rescued recipients compared with our overall transplantation population (276 +/- 86 min vs. 242 +/- 70 min, p < 0.01). The indication for transplantation, for ECMO support, and the timing of cannulation had no impact on survival. Hyperacute rejection was uncommon. Fifteen children were successfully weaned off ECMO and discharged alive (54%). Mean duration of ECMO was 2.8 days for survivors (median 3 days) compared with 4.8 days for nonsurvivors (median 5 days). There was 100% 3-year survival in the ECMO survivor group, with 13 patients (46%) currently alive at a mean follow-up of 8.1 +/- 3.8 years. The graft function was preserved (shortening fraction 36 +/- 7%), despite an increased number of early rejection episodes (1.7 +/- 1.6 vs. 0.7 +/- 1.3, overall transplant population, p < 0.05) and hemodynamically comprising rejection episodes (1.3 +/- 1.9 vs. 0.7 +/- 1.3, overall transplant population, p < 0.05). CONCLUSIONS: Overall survival was 54%, with all patients surviving to at least 3 years after undergoing transplantation. None of the children requiring >4 days of ECMO support survived. Despite an increased number of early and hemodynamically compromising rejections, the long-term graft function is similar to our overall transplantation population. PMID- 19679253 TI - Re: Outcome of extracorporeal membrane oxygenation for early primary graft failure after pediatric heart transplantation. PMID- 19679254 TI - Left ventricular pseudoaneurysm. PMID- 19679255 TI - Greatness. PMID- 19679256 TI - Advanced heart failure and transplant cardiology as a single subspecialty may restrict access to care. PMID- 19679258 TI - Most would fail to benefit from JUPITER Intervention. PMID- 19679260 TI - Coronary calcium scoring: calcium location needs to be integrated! PMID- 19679262 TI - Dietary fructose and intestinal barrier: potential risk factor in the pathogenesis of nonalcoholic fatty liver disease. AB - Worldwide, not only the prevalence of obesity has increased dramatically throughout the last three decades but also the incidences of co-morbid conditions such as diabetes type 2 and liver disease have increased. The 'hepatic manifestation of the metabolic syndrome' is called nonalcoholic fatty liver disease (NAFLD) and comprises a wide spectrum of stages of liver disease ranging from simple steatosis to liver cirrhosis. NAFLD of different stages is found in approximately 30% of adults and approximately 20% in the US population. Not just a general overnutrition but also an elevated intake of certain macronutrients such as fat and carbohydrates and herein particularly fructose has been claimed to be risk factors for the development for NAFLD; however, the etiology of this disease is still unknown. The present review outlines some of the potential mechanisms associated with the development of NAFLD and fructose intake with a particular focus on the role of the intestinal barrier functions. PMID- 19679264 TI - Macrophage migration inhibitory factor: a noncanonical chemokine important in atherosclerosis. AB - In the recent years, atherogenesis has increasingly been linked to inflammatory processes in the injured vessel wall. Recruitment and arrest of monocytes, T cells, and neutrophils via the concerted actions of multiple chemokines and their chemokine receptors have been the subject of intense research and are being appreciated as key events underlying atherosclerotic lesion formation and progression. The evolutionary conserved cytokine macrophage migration inhibitory factor (MIF) exhibits prominent proinflammatory and proatherogenic functions, and the latest findings on its chemotactic and chemokine-like properties imply MIF as a crucial drug target for the treatment of inflammatory diseases. In this review, the role of MIF in atherosclerosis and injury-induced neointima formation is discussed. We place an emphasis on its proinflammatory and chemokine-like functions in the context of underlying extra- and intracellular signaling mechanisms. These findings clearly distinguish MIF from other cytokines in atherosclerosis and justify the intensive search for inhibitors targeting MIF in the treatment of inflammatory diseases, including advanced atherosclerosis. PMID- 19679265 TI - Will the real plaque vasculature please stand up? Why we need to distinguish the vasa plaquorum from the vasa vasorum. AB - Many studies of experimental atherosclerosis and pathologic observations of human specimens have provided evidence supporting a correlation between vascularization of the atherosclerotic plaque and its natural growth and progression toward acute failure, associated with clinical events. The growing interest in the topic is illustrated by several excellent recent reviews discussing the molecular mechanisms that might play a role in the formation of plaque vasculature and that could explain some of the observed associations with pathologic features of experimental and human atherosclerotic lesions. At the same time, these reviews also emphasize that the field is still largely in uncharted territory. Hoping to spark some new investigations, we are taking this opportunity to question some of the common assumptions and to highlight less explored mechanisms. Finally, we are proposing to adopt the term vasa plaquorum to refer to the neovasculature located within the atherosclerotic plaque to distinguish it clearly from vasa vasorum, the native, supporting vasculature of the artery. We suggest that this new nomenclature offers a potential solution to eliminate ambiguity regarding implicit, but frequently neglected, differences between these structures. We think these points are relevant for future efforts to tailor diagnostic tools and therapeutic interventions targeting plaque neovascularization for the clinical management of atherosclerosis. PMID- 19679263 TI - Using new tools to define the genetic underpinnings of risky traits associated with coronary artery disease: the SardiNIA study. AB - Genomewide association studies are increasingly being applied to search for novel genes that might underlie cardiovascular diseases. In this article, we briefly review the principles that underlie modern genetic analyses and provide several illustrations from the SardiNIA study of genomewide association studies for cardiovascular risk factor traits. PMID- 19679266 TI - The clinical implications of the CHICAGO study for the management of cardiovascular risk in patients with type 2 diabetes mellitus. AB - The CHICAGO study (Carotid Intima-Media Thickness in Atherosclerosis Using Pioglitazone) tested the hypothesis that pioglitazone would have a beneficial effect for reducing CIMT progression, compared with glimepiride. Treatment with pioglitazone produced improvement in several parameters, such as systolic blood pressure and lipid levels, including a 14% increase in HDL cholesterol, and reduced CIMT progression, compared with glimepiride. However, only the beneficial effect on HDL cholesterol predicted its beneficial effect for reducing CIMT progression. The following review examines data from additional studies evaluating how pioglitazone and HDL levels modify cardiovascular risk. PMID- 19679267 TI - Novel insights of secretory phospholipase a(2) action in cardiology. AB - Phospholipase A(2)s (PLA(2)s) produce free fatty acid and lysophospholipid from membrane phospholipid, and these products can be converted into various types of proinflammatory lipid mediators by specific enzymes. Among several types of PLA(2), secretory PLA(2)s (sPLA(2)s) have crucial roles in the development of cardiovascular diseases. Circulating sPLA(2) is increasing in patients with coronary artery disease (CAD), and it can be a risk factor for CAD and a prognostic factor in those patients. Secretory PLA(2)s amplify the inflammatory responses in myocardial ischemia/reperfusion injury and fetal acute respiratory distress. In some animal experiments, sPLA(2)s can hydrolyze low-density lipoprotein and high-density lipoprotein and lead to progress of atherosclerotic plaques. Some inhibitor studies for sPLA(2) revealed that inhibition of sPLA(2) reduced the myocardial impairment after ischemia/reperfusion injury and progression of atherosclerotic plaque areas in animal models. Secretory PLA(2)s might be a new target for cardiovascular medicine. PMID- 19679268 TI - Hemoglobin-based red blood cell substitutes and nitric oxide. AB - Hemoglobin-based oxygen carriers (HBOCs) have been studied for decades as red blood cell substitutes. Profound vasoconstrictor effects have limited the clinical utility of HBOCs and are attributable to avid scavenging of nitric oxide (NO). Inhaling NO can charge the body's stores of NO metabolites without producing hypotension and can prevent systemic hypertension induced when HBOCs are subsequently infused. Concurrent breathing of low NO doses can prevent pulmonary vasoconstriction after HBOC infusion without augmenting plasma methemoglobinemia. PMID- 19679269 TI - Genetic testing for disease susceptibility: a risky business. PMID- 19679270 TI - Clinical trials and methodological problems in prion diseases. PMID- 19679272 TI - Bringing arts-based therapies in from the scientific cold. PMID- 19679274 TI - Emerging concepts in neural stem cell research: autologous repair and cell-based disease modelling. AB - The increasing availability of human pluripotent stem cells provides new prospects for neural-replacement strategies and disease-related basic research. With almost unlimited potential for self-renewal, the use of human embryonic stem cells (ESCs) bypasses the restricted supply and expandability of primary cells that has been a major bottleneck in previous neural transplantation approaches. Translation of developmental patterning and cell-type specification techniques to human ESC cultures enables in vitro generation of various neuronal and glial cell types. The derivation of stably proliferating neural stem cells from human ESCs further facilitates standardisation and circumvents the problem of batch-to-batch variations commonly encountered in "run-through" protocols, which promote terminal differentiation of pluripotent stem cells into somatic cell types without defined intermediate precursor stages. The advent of cell reprogramming offers an opportunity to translate these advances to induced pluripotent stem cells, thereby enabling the generation of neurons and glia from individual patients. Eventually, reprogramming could provide a supply of autologous neural cells for transplantation, and could lead to the establishment of cellular model systems of neurological diseases. PMID- 19679275 TI - Focal cortical dysplasia type II: biological features and clinical perspectives. AB - Focal cortical dysplasia (FCD) type II is an important cause of drug-resistant epilepsy. Clinical presentation is variable, and depends on age of onset of seizures and the location and size of lesion. As FCD type II cannot be diagnosed with certainty in the clinic, in vivo identification by use of MRI is important. Diagnosis will have a major effect on management of this pathology as it should prompt referral for specialist assessment. Drug treatment commonly proves ineffective, whereas appropriate surgical treatment can be curative in many cases. The dramatic cellular anomalies of FCD seen at histopathology indicate a widespread pattern of molecular disruption underpinning the structural disorganisation of the cortex. The cause for FCD has not been firmly established, and there are no explanations for its potent intrinsic ability to cause seizures. There seem to be both neurodevelopmental abnormalities and possible premature neurodegeneration in FCD. Understanding the coordination of the abnormal processes in FCD type II might help to promote improved detection in vivo, direct treatment strategies, and perhaps help explain the development, differentiation, and loss of brain cells, with broad implications for the epilepsies and other neurological disorders. PMID- 19679276 TI - Treatment of hyperkinetic movement disorders. AB - Parkinson's disease, the most common hypokinetic movement disorder, has received much attention from the clinical and scientific community, but there has been a relative paucity of comprehensive reviews of hyperkinetic disorders, even though they are equally or even more disabling. Hyperkinetic movement disorders include tremors, dystonia, chorea, tics, myoclonus, stereotypies, restless legs syndrome, and various other disorders with abnormal involuntary movements. Substantial progress has been made in the understanding of the role of the basal ganglia in the pathophysiology of these hyperkinesia disorders and in motor control, muscle tone, posture, and cognitive processes. Although therapies that target pathogenesis are still lacking, effective management of hyperkinetic movement disorders demands that physicians are knowledgeable about current and novel pharmacological and surgical approaches. In addition to tetrabenazine, a monoamine-depleting drug, new formulations of botulinum toxin are being increasingly used in the treatment of these movement disorders. Finally, success with surgical approaches, particularly deep brain stimulation in patients with Parkinson's disease who have levodopa-induced dyskinesias, has been extended to the treatment of many hyperkinetic movement disorders. PMID- 19679277 TI - Central post-stroke pain: clinical characteristics, pathophysiology, and management. AB - Central post-stroke pain (CPSP) is a neuropathic pain syndrome that can occur after a cerebrovascular accident. This syndrome is characterised by pain and sensory abnormalities in the body parts that correspond to the brain territory that has been injured by the cerebrovascular lesion. The presence of sensory loss and signs of hypersensitivity in the painful area in patients with CPSP might indicate the dual combination of deafferentation and the subsequent development of neuronal hyperexcitability. The exact prevalence of CPSP is not known, partly owing to the difficulty in distinguishing this syndrome from other pain types that can occur after stroke (such as shoulder pain, painful spasticity, persistent headache, and other musculoskeletal pain conditions). Future prospective studies with clear diagnostic criteria are essential for the proper collection and processing of epidemiological data. Although treatment of CPSP is difficult, the most effective approaches are those that target the increased neuronal hyperexcitability. PMID- 19679278 TI - Determinants of occurrence of aortic sclerosis in an aging population. AB - OBJECTIVES: We sought to identify clinical, physiological, and biochemical correlates, including markers of endothelial dysfunction and of tissue nitric oxide (NO) responsiveness, of the presence of aortic sclerosis (ASc) in an aging population. BACKGROUND: Aortic sclerosis has been regarded predominantly as a precursor of hemodynamically significant aortic stenosis. However, ASc also represents an independent correlate of increased risk of cardiovascular morbidity and mortality; the basis of this association is incompletely understood. The assumption that the pathogenesis of aortic valve disease is similar to that of atherosclerosis has not been supported by recent studies; rather there has been increasing evidence of a pathogenetic role of inflammation and endothelial dysfunction. Furthermore, we have recently developed methodology for echocardiographic quantitation of early aortic valve disease. METHODS: Randomly selected subjects (n = 253) ages 51 to 77 years underwent transthoracic echocardiography; aortic valve ultrasonic backscatter score (AV(BS)) was used to quantitate echogenicity of the aortic valve. Conventional coronary risk factors were identified. Integrity of NO generation/response was assessed via: 1) plasma asymmetric dimethylarginine concentrations, as a marker of endothelial dysfunction; 2) inhibition of platelet aggregation by the NO donor sodium nitroprusside, as a measure of tissue NO responsiveness and also a coronary prognostic marker; and 3) aortic augmentation index, as a measure of arterial stiffness/wave reflection. All putative correlations with AV(BS) were examined by univariate and multiple linear regression analyses. RESULTS: On the basis of AV(BS) scores, ASc was present in 19.4% of subjects. The AV(BS) directly correlated with patients' age but inversely correlated with high-sensitivity C reactive protein, creatinine clearance, and platelet NO responsiveness. On multiple linear regression, ASc was associated with impaired platelet NO responsiveness (beta = -0.16, p = 0.02), advancing age (beta = 0.21, p = 0.003), and low body mass index (beta = -0.23, p = 0.001). CONCLUSIONS: Aortic sclerosis is associated with platelet NO resistance rather than conventional coronary risk factors: this might explain the increased thrombotic risk in ASc. PMID- 19679279 TI - Is it time for a new paradigm in calcific aortic valve disease? PMID- 19679280 TI - Mechanisms of recurrent aortic regurgitation after aortic valve repair: predictive value of intraoperative transesophageal echocardiography. AB - OBJECTIVES: The aim of the present study was to examine the intraoperative echocardiographic features associated with recurrent severe aortic regurgitation (AR) after an aortic valve repair surgery. BACKGROUND: Surgical valve repair for AR has significant advantages over valve replacement, but little is known about the predictors and mechanisms of its failure. METHODS: We blindly reviewed all clinical, pre-operative, intraoperative, and follow-up transesophageal echocardiographic data of 186 consecutive patients who underwent valve repair for AR during a 10-year period and in whom intraoperative and follow-up echo data were available. After a median follow-up duration of 18 months, 41 patients had recurrent 3+ AR, 23 patients presented with residual 1+ to 2+ AR, and 122 had no or trivial AR. In patients with recurrent 3+ AR, the cause of recurrent AR was the rupture of a pericardial patch in 3 patients, a residual cusp prolapse in 26 patients, a restrictive cusp motion in 9 patients, an aortic dissection in 2 patients, and an infective endocarditis in 1 patient. RESULTS: Pre-operatively, all 3 groups were similar for aortic root dimensions and prevalence of bicuspid valve (overall 37%). Patients with recurrent AR were more likely to display Marfan syndrome or type 3 dysfunction pre-operatively. At the opposite end, patients with continent AR repair at follow-up were more likely to have type 2 dysfunction pre-operatively. After cardiopulmonary bypass, a shorter coaptation length, the degree of cusp billowing, a lower level of coaptation (relative to the annulus), a larger diameter of the aortic annulus and the sino-tubular junction, the presence of a residual AR, and the width of its vena contracta were associated with the presence of AR at follow-up. Multivariate Cox analysis identified a shorter coaptation length (odds ratio [OR]: 0.8, p = 0.05), a coaptation occurring below the level of the aortic annulus (OR: 7.9, p < 0.01), a larger aortic annulus (OR: 1.2, p = 0.01), and residual aortic regurgitation (OR: 5.3, p = 0.01) as risk factors of repair failure. CONCLUSIONS: Our results demonstrate that intraoperative transesophageal echocardiography can be used to identify patients undergoing AR repair who are at increased risk for late repair failure. PMID- 19679281 TI - Impact of a reduced tube voltage on CT angiography and radiation dose: results of the PROTECTION I study. AB - OBJECTIVES: The aim of this study was to determine the impact of a reduced 100-kV tube voltage on image quality and radiation exposure in a pre-defined subgroup analysis of the international, multicenter radiation dose survey PROTECTION I (Prospective Multicenter Study on RadiaTion Dose Estimates Of Cardiac CT AngIOgraphy I) study. BACKGROUND: Cardiac computed tomography angiography (CCTA) has become a frequently used diagnostic tool in clinical practice. Despite continually improving CT technology, there remain concerns regarding the associated radiation exposure. A reduced tube voltage of 100 kV has been proposed as an effective means for dose reduction in nonobese patients. METHODS: The study assessed the relevant radiation dose parameters as well as quantitative and qualitative diagnostic image quality data in a subgroup of 321 patients (100 kV: 82 patients; 120 kV: 239 patients), who were scanned at study sites that applied a 100-kV tube voltage in at least 1 patient. Diagnostic image quality was assessed by an experienced CCTA investigator with a 4-point score (1: nondiagnostic to 4: excellent image quality). Effective radiation dose was estimated from the dose-length-product of each CCTA study. RESULTS: The use of the 100-kV scan protocol was associated with 53% reduction in CCTA median radiation dose estimates, when compared with the conventional 120-kV scan protocol (p < 0.001). Although image noise significantly increased by 26.3% with 100 kV, signal- as well as contrast-to-noise ratios also increased by 7.9% (p = 0.254) and 10.8% (p = 0.027), respectively. Reduction of tube voltage did not impair diagnostic image quality (median diagnostic score: 3.5 [3.25 to 3.75] vs. 3.5 [3.0 to 3.75] for 100 kV vs. 120 kV; p = 0.22). CONCLUSIONS: In this nonrandomized PROTECTION I dose survey, reducing the CCTA tube voltage to 100 kV in nonobese patients is associated with a significant reduction in radiation exposure while maintaining diagnostic image quality. Thus, the 100-kV scan technique should be considered for CCTA dose reduction in adequately selected patients. PMID- 19679282 TI - CT Angiography by reduced tube voltage more than a single step. PMID- 19679283 TI - Characterization of complex coronary artery stenosis morphology by coronary computed tomographic angiography. AB - OBJECTIVES: This study sought to assess the ability of coronary computed tomography angiography (CTA) in identifying complex coronary stenosis morphology before invasive coronary angiography (ICA) and percutaneous coronary intervention (PCI). BACKGROUND: Complexity of stenosis morphology affects PCI success. Whether CTA can detect the entire spectrum of recognized complex stenosis morphologies has not been investigated. METHODS: All nonbypassed, nonstented, >or=2-mm diameter native coronary arterial segments in 85 consecutive patients who underwent ICA or=70% stenotic by visual inspection and characterized each as type C or nontype C, according to the modified American College of Cardiology morphology criteria for estimating PCI risk. Results were compared with ICA data similarly analyzed by 2 blinded interventional cardiologists. The PCI procedure duration and contrast use were compared between type C and nontype C lesions identified on both ICA and CTA. RESULTS: CTA detected 84 of 93 lesions (90%) causing >or=70% stenosis on ICA and correctly characterized 42 of 53 lesions (79%) found to concurrently show type C morphology on ICA. Type C features most frequently missed by CTA were ostial involvement (5 cases) and lesion length >20 mm (7 cases). Major branch involvement was the most frequent false-positive type C feature (12 cases). Mean PCI duration in patients with and without type C lesions on CTA were 42.4 +/- 24.7 min and 21.5 +/- 13.3 min (p = 0.009), respectively; mean total contrast used were 263 +/- 150 ml and 140 +/- 47 ml (p = 0.007), respectively. CONCLUSIONS: In vessels segments >or=2 mm in diameter, CTA can predict lesions likely to reach >or=70% stenosis on ICA and provide added value in discerning complex morphologies associated with these lesions. Presence of complex, severely obstructive lesions on CTA is associated with higher contrast use and greater procedure length during PCI. PMID- 19679284 TI - Regadenoson induces comparable left ventricular perfusion defects as adenosine: a quantitative analysis from the ADVANCE MPI 2 trial. AB - OBJECTIVES: This study sought to determine whether regadenoson induces left ventricular perfusion defects of similar size and severity as seen with adenosine stress. BACKGROUND: Total and ischemic left ventricular perfusion defect size predict patient outcome. Therefore, it is important to show that newer stressor agents induce similar perfusion abnormalities as observed with currently available ones. METHODS: The ADVANCE MPI 2 (Adenosine versus Regadenoson Comparative Evaluation for Myocardial Perfusion Imaging) study was a prospective, double-blind, randomized trial comparing image results in patients undergoing standard gated adenosine single-photon emission computed tomography (SPECT) myocardial perfusion imaging who were then randomized in a 2:1 ratio to either regadenoson (N = 495) or a second adenosine SPECT (N = 260). Quantitative SPECT analysis was used to determine total left ventricular perfusion defect size and the extent of ischemia. Quantification was performed by a single observer who was blinded to randomization and image sequence. RESULTS: Baseline gated perfusion results were similar in patients randomized to adenosine or regadenoson. No significant differences in total (11.5 +/- 15.7 vs. 11.4 +/- 15.8, p = 0.88) or ischemic (4.8 +/- 9.2 vs. 4.6 +/- 8.9, p = 0.43) perfusion defect sizes were observed between the regadenoson and adenosine groups, respectively. Linear regression showed a close correlation between adenosine and regadenoson for total (r = 0.97, p < 0.001) and ischemic (r = 0.95, p < 0.001) left ventricular perfusion defects. Serial differences in total (-0.03 +/- 3.89 vs. -0.13 +/- 4.16, p = 0.73) and ischemic (0.15 +/- 4.08 vs. 0.25 +/- 3.81, p = 0.74) perfusion defect size and left ventricular ejection fraction (0.12 +/- 0.32 vs. 0.15 +/- 0.35, p = 0.27) from study 1 to study 2 were virtually identical in patients randomized to regadenoson versus adenosine, respectively. The good correlation between serial adenosine and regadenoson studies regarding total (0.41 +/- 5.43 vs. 0.21 +/- 5.23, p = 0.76) and ischemic (0.17 +/- 5.31 vs. 0.23 +/- 6.08, p = 0.94) perfusion defects persisted in the subgroup of 308 patients with an abnormal baseline SPECT. CONCLUSIONS: Applying quantitative analysis, regadenoson induces virtually identical scintigraphic results as adenosine regarding the size and severity of left ventricular perfusion defects and the extent of scintigraphic ischemia. PMID- 19679285 TI - Contrast-enhanced anatomic imaging as compared to contrast-enhanced tissue characterization for detection of left ventricular thrombus. AB - OBJECTIVES: This study sought to compare contrast-enhanced anatomic imaging and contrast-enhanced tissue characterization (delayed-enhancement cardiac magnetic resonance [DE-CMR]) for left ventricular (LV) thrombus detection. BACKGROUND: Contrast echocardiography (echo) detects LV thrombus based on anatomic appearance, whereas DE-CMR imaging detects thrombus based on tissue characteristics. Although DE-CMR has been validated as an accurate technique for thrombus, its utility compared with contrast echo is unknown. METHODS: Multimodality imaging was performed in 121 patients at high risk for thrombus due to myocardial infarction or heart failure. Imaging included 3 anatomic imaging techniques for thrombus detection (contrast echo, noncontrast echo, cine-CMR) and a reference of DE-CMR tissue characterization. LV structural parameters were quantified to identify markers for thrombus and predictors of additive utility of contrast-enhanced thrombus imaging. RESULTS: Twenty-four patients had thrombus by DE-CMR. Patients with thrombus had larger infarcts (by DE-CMR), more aneurysms, and lower LV ejection fraction (by CMR and echo) than those without thrombus. Contrast echo nearly doubled sensitivity (61% vs. 33%, p < 0.05) and yielded improved accuracy (92% vs. 82%, p < 0.01) versus noncontrast echo. Patients who derived incremental diagnostic utility from DE-CMR had lower LV ejection fraction versus those in whom noncontrast echo alone accurately assessed thrombus (35 +/- 9% vs. 42 +/- 14%, p < 0.01), with a similar trend for patients who derived incremental benefit from contrast echo (p = 0.08). Contrast echo and cine-CMR closely agreed on the diagnosis of thrombus (kappa = 0.79, p < 0.001). Thrombus prevalence was lower by contrast echo than DE-CMR (p < 0.05). Thrombus detected by DE-CMR but not by contrast echo was more likely to be mural in shape or, when apical, small in volume (p < 0.05). CONCLUSIONS: Echo contrast in high-risk patients markedly improves detection of LV thrombus, but does not detect a substantial number of thrombi identified by DE-CMR tissue characterization. Thrombi detected by DE-CMR but not by contrast echo are typically mural in shape or small in volume. PMID- 19679286 TI - Noncardiac pathology on clinical cardiac magnetic resonance imaging. AB - OBJECTIVES: We sought to determine the prevalence of noncardiac pathology in a large consecutive series of patients referred for clinical cardiac magnetic resonance (CMR) studies. BACKGROUND: The imaging field for many CMR sequences extends outside of the heart border. As a result, noncardiac pathology may be identified. These noncardiac findings have clinical significance because they often lead to subsequent imaging/testing and intervention. The prevalence of noncardiac findings on clinical CMR studies has not been well described. METHODS: The reports of all 1,534 (62% male, age 50 +/- 15 years) clinical CMR studies performed at an academic medical center during calendar years 2002 to 2006 were reviewed. All studies had been interpreted by both a staff cardiologist (level III trained in CMR) and a board-certified radiologist (with fellowship training in CMR). For each study, sex, age, indication for CMR study, and reported noncardiac pathology were extracted. Follow-up for each major noncardiac pathology was evaluated by reviewing the patient's medical center electronic medical record. These noncardiac pathologies were then categorized as significant if an intervention or change in the patient's management ensued. RESULTS: A total of 116 (7.6%) studies had at least one noncardiac finding. These findings included 55 major findings (e.g., lymphadenopathy, lung abnormalities, mediastinal masses) in 48 distinct reports (prevalence of 3.1%) and 74 minor findings (e.g., small pleural effusions, liver cysts, renal cysts) in 70 distinct reports (prevalence of 4.6%). The majority (62%) of major findings were previously known, with only 8 findings in 6 (0.4%) of 1,534 reports ultimately deemed to be new and clinically important/significant. The age of those with noncardiac pathology was greater (54 +/- 16 years vs. 49 +/- 16 years, p < 0.001). CONCLUSIONS: In this large series of consecutive clinical CMR studies interpreted by both staff cardiologists and radiologists, noncardiac pathology is uncommonly reported. When reported, the majority of major findings are previously known. New major findings were detected in <0.5% of reports. PMID- 19679287 TI - Molecular MRI of early thrombus formation using a bimodal alpha2-antiplasmin based contrast agent. AB - OBJECTIVES: We aimed to investigate whether early thrombus formation can be visualized with in vivo magnetic resonance imaging (MRI) by the use of a novel bimodal alpha(2)-antiplasmin-based contrast agent (CA). BACKGROUND: Thrombus formation plays a central role in several vascular diseases. During the early phases of thrombus formation, activated factor XIII (FXIIIa) covalently cross links alpha(2)-antiplasmin to fibrin, indicating the potential of alpha(2) antiplasmin-based CAs in the detection of early thrombus formation. METHODS: A bimodal CA was synthesized by coupling gadolinium-diethylene triamine pentaacetic acid and rhodamine to an alpha(2)-antiplasmin-based peptide. For the control CA, a glutamine residue essential for cross-linking was replaced by alanine. In vitro generated thrombi were exposed to both CAs and imaged by MRI and 2-photon laser scanning microscopy. Immunohistochemistry was performed on human pulmonary thromboemboli sections to determine the presence of alpha(2)-antiplasmin and FXIII in different thrombus remodeling phases. In vivo feasibility of the CA in detecting early thrombus formation specifically was investigated with MRI. RESULTS: In vitro-generated thrombi exposed to the alpha(2)-antiplasmin-based CA showed hyperintense magnetic resonance signal intensities at the thrombus edge. No hyperintense signal was observed when we used the alpha(2)-antiplasmin-based CA in the presence of FXIII inhibitor dansylcadaverine nor when we used the control CA. Two-photon laser-scanning microscopy demonstrated that the alpha(2) antiplasmin-based CA bound to fibrin. Immunohistochemistry demonstrated substantial alpha(2)-antiplasmin staining in fresh compared with lytic and organized thrombi. The administration of CA in vivo within seconds after inducing thrombus formation increased contrast-to-noise ratios (CNRs 2.28 +/- 0.39, n=6) at the site of thrombus formation compared with the control CA (CNRs -0.14 +/- 0.55, p = 0.003, n = 6) and alpha(2)-antiplasmin-based CA administration 24 to 48 h after thrombus formation (CNRs 0.11 +/- 0.23, p = 0.006, n = 6). CONCLUSIONS: A bimodal CA was developed, characterized, and validated. Our results showed that this bimodal CA enabled noninvasive in vivo magnetic resonance visualization of early thrombus formation. PMID- 19679288 TI - Specific targeting of human inflamed endothelium and in situ vascular tissue transfection by the use of ultrasound contrast agents. AB - OBJECTIVES: We used human umbilical cord segments as an ex vivo model to investigate the possible clinical diagnostic and therapeutic applications of microbubbles (MBs). BACKGROUND: Microbubbles are commonly used in clinical practice as ultrasound contrast agents. Several studies have addressed the in vivo applications of MBs for specific targeting of vascular dysfunction or sonoporation in animal models, but to date no human tissue model has been established. METHODS: Primary venular endothelial cell monolayers were targeted with MBs conjugated to an antibody against a highly expressed endothelial marker (tetraspanin CD9), and binding was assessed under increasing flow rates (0.5 to 5 dynes/cm(2)). Furthermore, CD9-coupled MB endothelial targeting was measured under flow conditions by contrast-enhanced ultrasound analysis in an ex vivo human macrovascular model (umbilical cord vein), and the same tissue model was used for the detection of inflamed vasculature with anti-intercellular adhesion molecule (ICAM)-1-coated MBs. Finally, plasmids encoding fluorescent proteins were sonoporated into umbilical cord vessels. RESULTS: Specific endothelial targeting in the in vitro and ex vivo models described previously was achieved by the use of MBs covered with an anti-CD9. Furthermore, we managed to induce inflammation in umbilical cord veins and detect it with real-time echography imaging using anti-ICAM-1-coupled MBs. Moreover, expression and correct localization of green fluorescent protein and green fluorescent protein-tagged ICAM-1 were assessed in this human ex vivo model without causing vascular damage. CONCLUSIONS: In the absence of clinical trials to test the benefits and possible applications of ultrasound contrast agents for molecular imaging and therapy, we have developed a novel ex vivo human model using umbilical cords that is valid for the detection of inflammation and for exogenous expression of proteins by sonoporation. PMID- 19679289 TI - The umbilical cord: an ally in targeted imaging research? PMID- 19679290 TI - Does coronary flow trump coronary anatomy? AB - Coronary function versus anatomy, flow versus stenosis: which optimizes coronary artery disease (CAD) management? In patients, coronary flow is poorly related to stenosis severity, and revascularization fails to improve mortality over medical treatment in randomized trials. Yet percutaneous intervention (PCI) guided by fractional flow reserve reduces coronary events more than PCI guided by arteriographic stenosis. These paradoxes are explained by the poor relation between coronary flow reserve (CFR) and stenosis severity due to diffuse CAD, with surprising clinical implications. Should the concept of anatomically "critical" coronary stenosis be replaced by the concept of "critical" CFR reduction for managing CAD? PMID- 19679291 TI - Correction of pulmonary arteriovenous malformation using image-based surgical planning. AB - The objectives of this study were to develop an image-based surgical planning framework that 1) allows for in-depth analysis of pre-operative hemodynamics by the use of cardiac magnetic resonance and 2) enables surgeons to determine the optimum surgical scenarios before the operation. This framework is tailored for applications in which post-operative hemodynamics are important. In particular, it is exemplified here for a Fontan patient with severe left pulmonary arteriovenous malformations due to abnormal hepatic flow distribution to the lungs. Patients first undergo cardiac magnetic resonance for 3-dimensional anatomy and flow reconstruction. After analysis of the pre-operative flow fields, the 3-dimensional anatomy is imported into an interactive surgical planning interface for the surgeon to virtually perform multiple surgical scenarios. Associated hemodynamics are predicted by the use of a fully validated computational fluid dynamic solver. Finally, efficiency metrics (e.g., pressure decrease and hepatic flow distribution) are weighted against surgical feasibility to determine the optimal surgical option. PMID- 19679292 TI - Real-time 3-dimensional transesophageal echocardiography during double percutaneous mitral edge-to-edge procedure. PMID- 19679293 TI - 3D TEE for evaluation of commissural opening before and during percutaneous mitral commissurotomy. PMID- 19679295 TI - Qualitative and quantitative scrutiny by regulatory process: is the truth subjective or objective? PMID- 19679296 TI - Methylation frequencies of cell-cycle associated genes in epithelial odontogenic tumours. AB - OBJECTIVE: The benign epithelial odontogenic tumours constitute a group of lesions derived from epithelial elements of the tooth-forming apparatus. This group includes lesions of different biological behaviour, such as ameloblastoma, calcifying cystic odontogenic tumour (CCOT) and adenomatoid odontogenic tumour (AOT). The pathogenesis of these neoplasms remains uncertain and the occurrence of methylation in cell-cycle related genes may be involved in their development. The aim of this study was to investigate the methylation status of P16, P21, P27, P53 and RB1 genes in epithelial odontogenic tumours. DESIGN: Methylation-specific polymerase chain reaction (MSP) was used to evaluate the presence of methylation in 13 samples of ameloblastoma, six samples of CCOT, three samples of AOT and 14 samples of dental follicles, included as control. RESULTS: Our results showed a distinct methylation profile in each group. In ameloblastoma, the highest methylated genes were P16 and P21, while in CCOT the P21 and RB1 genes were the most commonly methylated genes. Only the P16 and P21 genes were methylated in the AOT samples. In the dental follicle samples, P16, P27 and RB1 genes were commonly methylated. A high percentage of the odontogenic tumours analysed showed methylation of the P21 gene, in contrast to dental follicles. CONCLUSIONS: Epithelial odontogenic tumours show a distinct methylation profile in cell-cycle associated genes. In addition to this, the current findings show that epigenetic alterations are common events in epithelial odontogenic tumours. PMID- 19679297 TI - Cyclin D1 expression and its possible regulation in chewing tobacco mediated oral squamous cell carcinoma progression. AB - OBJECTIVE: Proto-oncogene cyclin D1 is a G1 phase specific cell cycle regulator and known for its role in various cancers. The aim of the study was to understand oral cancer progression by observing the mRNA and protein expression of cyclin D1. METHODS: Different oral tissue samples were selected as a model to study oral cancer progression. Those include healthy oral mucosa, premalignant lesions (Leukoplakia, Erythroplakia, Oral SubMucous Fibrosis) and oral cancer (OSCC) samples. Cyclin D1 mRNA and protein expression were detected by slot-blot and by immunohistochemical methods, respectively. RESULTS: Premalignant lesions (PML) showed average 3-fold increase in the mRNA expression than normal oral mucosa (p = 0.001) whereas only 1.3-fold increase in mRNA has been observed in OSCC samples over the PML. On the other hand OSCC showed average 4-fold increase in mRNA expression than normal oral mucosa (p < 0.001). Cyclin D1 protein accumulation has been observed in 31.3% (16/51) of the OSCC samples whereas the normal oral mucosa and the PML showed no immunoreactivity. Oral cancer samples showing positive cyclin D1 immunoreactivity has increased from 15.0% (3/20) well differentiated SCC to 31.2% (5/16) moderately differentiated SCC to 53.3% (8/15) poorly differentiated SCC, found statistically significant (p = 0.05). CONCLUSION: By observing the expression of cyclin D1 in different stages, we have noticed two major transitions that occur in normal oral mucosa that leads to oral cancer. The first transitional event transforms the normal oral mucosa to PML whereas the second transition drives the PML to OSCC. These findings give evidence that the first transition induces cyclin D1 mRNA with no detectable cyclin D1 protein. The induction of mRNA is maintained with increased cyclin D1 protein accumulation in the second transition. PMID- 19679298 TI - Comparing visual and auditory presentation for the modification of interpretation bias. AB - This experiment compares the effects of visual vs. auditory presentation of cognitive bias modification (CBM) training scenarios upon interpretation style and emotional vulnerability. For both modalities, negative, but not positive interpretation biases were successfully induced relative to a baseline. Mood declined for the auditory but not the visual group throughout the CBM procedure, irrespective of the valence of the CBM condition. This deterioration in mood raises an important methodological issue and indicates that the increased testing time brought about by auditory compared to visual presentation needs to be addressed. The CBM procedures did not influence emotional vulnerability as assessed by behavioural measures, but counter-to-prediction, CBM procedures did increase self-reported depression vulnerability for the positive but not negative CBM condition. PMID- 19679299 TI - Experimental modification of interpretation bias in socially anxious children: Changes in interpretation, anticipated interpersonal anxiety, and social anxiety symptoms. AB - We report on an experimental manipulation of interpretation bias in socially anxious youths. A non-clinical sample of 10-11-year-olds selected for high social anxiety was trained over three sessions to endorse benign rather than negative interpretations of potentially threatening social scenarios. This group was subsequently less likely to endorse negative interpretations of new ambiguous social situations than children in a test-retest condition. Children who received interpretation training also showed reduced trait social anxiety and reported significantly less anxiety about an anticipated interpersonal encounter, compared with the control group. PMID- 19679300 TI - Ost's (2008) methodological comparison of clinical trials of acceptance and commitment therapy versus cognitive behavior therapy: Matching apples with oranges? AB - Ost (2008) recently compared the methodological rigor of studies of acceptance and commitment therapy (ACT) and traditional cognitive behavior therapy (CBT). He concluded that the ACT studies had more methodological deficiencies, and thus the treatment did not qualify as an "empirically supported treatment." Although Ost noted several important limitations that should be carefully considered when evaluating early ACT research, his attempt to devise an empirical matching strategy by creating a comparison sample of CBT studies to bolster his conclusions was itself problematic. The samples were clearly mismatched in terms of the populations being treated, leading to differences in study design and methodology. Furthermore, reanalysis showed clear differences in grant support favoring CBT compared with ACT studies that were not reported in the original article. Given the actual mismatch between the samples, Ost's methodological ratings are difficult to interpret and provide little useful information beyond what could already be gathered by a qualitative review of ACT study limitations. Such limitations are characteristic of the earlier randomized controlled trials of any emerging psychotherapeutic approach. PMID- 19679301 TI - Understanding the relationship between posttraumatic stress disorder and trauma cognitions: The impact of thought control strategies. AB - Several models of PTSD suggest that dysfunctional beliefs play an important role in the maintenance of PTSD. This study examined whether thought control strategies intermediated in the relationship between PTSD and dysfunctional cognitions. It was hypothesized that PTSD would be positively associated with dysfunctional cognitions and that negative thought control strategies (worry and self-punishment) would partially account for this relationship. These maladaptive strategies were predicted to be associated with increased levels of PTSD and more trauma-related beliefs. Additionally, it was predicted that positive thought control strategies (social control and reappraisal) would be associated with decreased levels of PTSD and fewer trauma-related beliefs. Finally, because the literature supports distraction as both an adaptive and a maladaptive thought control strategy, no a priori hypothesis was made. Results support worry and self punishment as maladaptive intervening variables in the association between PTSD and dysfunctional cognitions, resulting in greater levels of PTSD and trauma cognitions. Social control and distraction emerged as adaptive strategies, resulting in lower levels of PTSD and trauma cognitions, while reappraisal showed no relationship with PTSD severity. Although the results are cross-sectional, continued focus on the effects of thought control strategies as meditational maintenance variables over time appears warranted. PMID- 19679302 TI - Predicting anger in social anxiety: The mediating role of rumination. AB - Anger and the way that anger is expressed have been linked to attrition and poorer treatment outcomes in patients with Social Anxiety Disorder (e.g., Erwin, B. A., Heimberg, R. G., Schneier, F. R. & Liebowitz, M. R. (2003). Anger experience and expression in social anxiety disorder: Pretreatment profile and predictors of attrition and response to cognitive-behavioral treatment. Behavior Therapy, 34, 331-350). Understanding the connection between social anxiety and anger may be one way to improve outcomes in this population. A cross-sectional regression design was used in a sample of 363 undergraduates to examine the suggestion that ruminative thought is a critical factor linking social anxiety to anger. In support of this hypothesis, brooding fully mediated the relationship between social anxiety and trait anger and partially mediated the relationship between social anxiety and outward anger expression. The relationship between brooding and anger suppression became non-significant after depression was controlled. In contrast, reflective pondering partially mediated the relationship between social anxiety and anger suppression. These results suggest that addressing rumination may be useful in the treatment of socially anxious patients who struggle with anger. They also support the utility of considering multiple forms of rumination and multiple anger outcomes in a single study. PMID- 19679303 TI - An apigenin-induced decrease in K-antigen production by Sinorhizobium sp. NGR234 is y4gM- and nodD1-dependent. AB - Cultured cells of Sinorhizobium sp. NGR234 produce an abundance of capsular polysaccharides, or K antigens; however, cells that are cultured in the presence of apigenin, a nod gene inducer, exhibited a significant reduction in K-antigen production. The flavonoid-induced modulation in capsule production appeared to be related to the phase-shift changes associated with bacteroid differentiation. Therefore, the polysaccharides were extracted from Sinorhizobium sp. NGR234 bacteroids recovered from Vigna unguiculata cv Red Caloona root nodules, and subsequent analyses showed that the bacteroid extracts were virtually devoid of K antigen. Polysaccharide extracts from two nodulation mutants cultured in the presence of apigenin were then analyzed, and the results showed that the flavonoid-inducible decrease in K-antigen production is y4gM- and nodD1 dependent. PMID- 19679304 TI - Reliability and validity of the Japanese version of the Recovery Assessment Scale (RAS) for people with chronic mental illness: scale development. AB - BACKGROUND: Recovery is defined as a complex process of developing new meaning and purpose in life as one grows beyond the catastrophic effects of mental illness. To promote this process, the necessity of recovery assessment using psychometric measures has been emphasized; however, no measure to assess the individual recovery process is available in Japan. OBJECTIVE: To develop a Japanese version of the Recovery Assessment Scale (RAS) and to examine its reliability and validity. DESIGN: The study was a cross-sectional questionnaire survey. SETTINGS: Participants came from 1 daycare, 1 outpatient clinic of a psychiatric hospital, 5 sheltered workshops used for social rehabilitation, 3 peer support groups, and 6 inpatient wards of 2 psychiatric hospitals. PARTICIPANTS: The survey included 237 participants who had chronic mental illness and were aged 20 or older. For analysis, we used data from 209 participants who had no missing values on the RAS, with 58.9% male and a mean age of 48.3 years. METHODS: The questionnaire consisted of the Japanese version of the 24-item RAS, developed by the authors with focus group cognitive interviews and the translation-back-translation procedure. Also included were the Herth Hope Index, Empowerment scale, Resilience scale, SF-8, and BASIS-32. Internal consistency reliability was assessed by Cronbach's alpha coefficients, and test-retest reliability was assessed by the intraclass correlation coefficient (ICC) and weighted kappa in a randomly selected subsample (n=24). Exploratory and confirmatory factor analyses and correlations with other scales were used to examine the factor-based validity, concurrent and construct validity of the RAS. RESULTS: Cronbach's alpha coefficient was 0.89 for the overall RAS. ICC and weighted kappa generally indicated good test-retest reliability. Factor analysis of the RAS items yielded five factors: (a) goal/success orientation and hope, (b) reliance on others, (c) personal confidence, (d) no domination by symptoms, and (e) willingness to ask for help. The item "Coping with mental illness is no longer the main focus of my life" showed an inverse factor loading. The overall RAS score significantly and positively correlated with the Herth Hope Index, Empowerment scale, Resilience scale, and SF-8 mental component summary; there was a significant negative correlation with BASIS-32 psychiatric symptoms and functional impairment (p<0.01). CONCLUSION: This study confirmed the reliability and validity of the Japanese version of the 24-item RAS among people with chronic mental illness currently living in communities and inpatient ward settings in Japan. PMID- 19679305 TI - Mast cell-dependent proteolytic modification of HDL particles during anaphylactic shock in the mouse reduces their ability to induce cholesterol efflux from macrophage foam cells ex vivo. AB - OBJECTIVE: We have found previously that proteolytic modification of HDL by mast cell chymase in vitro reduces cholesterol efflux from cultured macrophage foam cells. Here, we evaluated whether mast cell-dependent proteolysis of HDL particles may occur in vivo, and whether such modification would impair their function in inducing cellular cholesterol efflux ex vivo. METHODS: Systemic activation of mast cells in the mouse was achieved by intraperitoneal injection of a high dose of the mast cell-specific noncytotoxic degranulating agent, compound 48/80. Serum and intraperitoneal fluid were then evaluated for degradation of HDL apolipoproteins and for their potential to act as cholesterol acceptors from cultured mouse macrophage foam cells. RESULTS: Lysates of isolated mouse peritoneal mast cells containing active chymase partially proteolyzed apoA I in alpha- and prebeta-HDL particles in mouse serum in vitro, and, when injected into the mouse peritoneal cavity, the lysates also degraded endogenous apoA-I in peritoneal fluid in vivo. Systemic activation of mast cells in mast cell competent mice, but not in mast cell-deficient (W-sash c-kit mutant) mice, reduced the ability of serum and intraperitoneal fluid derived from these animals to promote efflux of cellular cholesterol. This inhibitory effect was related to mast cell-dependent proteolytic degradation of apoA-I, apoA-IV, and apoE, i.e., the HDL-associated apolipoproteins that are efficient inducers of cholesterol efflux. CONCLUSION: The present results document a role for extracellular mast cell-dependent proteolysis in the generation of dysfunctional HDL, and suggest an inhibitory role for mast cells in the initial step of reverse cholesterol transport in vivo. PMID- 19679306 TI - The effects of sterol structure upon sterol esterification. AB - Cholesterol is esterified in mammals by two enzymes: LCAT (lecithin cholesterol acyltransferase) in plasma and ACAT(1) and ACAT(2) (acyl-CoA cholesterol acyltransferases) in the tissues. We hypothesized that the sterol structure may have significant effects on the outcome of esterification by these enzymes. To test this hypothesis, we analyzed sterol esters in plasma and tissues in patients having non-cholesterol sterols (sitosterolemia and Smith-Lemli-Opitz syndrome). The esterification of a given sterol was defined as the sterol ester percentage of total sterols. The esterification of cholesterol in plasma by LCAT was 67% and in tissues by ACAT was 64%. Esterification of nine sterols (cholesterol, cholestanol, campesterol, stigmasterol, sitosterol, campestanol, sitostanol, 7 dehydrocholesterol and 8-dehydrocholesterol) was examined. The relative esterification (cholesterol being 1.0) of these sterols by the plasma LCAT was 1.00, 0.95, 0.89, 0.40, 0.85, 0.82 and 0.80, 0.69 and 0.82, respectively. The esterification by the tissue ACAT was 1.00, 1.29, 0.75, 0.49, 0.45, 1.21 and 0.74, respectively. The predominant fatty acid of the sterol esters was linoleic acid for LCAT and oleic acid for ACAT. We compared the esterification of two sterols differing by only one functional group (a chemical group attached to sterol nucleus) and were able to quantify the effects of individual functional groups on sterol esterification. The saturation of the A ring of cholesterol increased ester formation by ACAT by 29% and decreased the esterification by LCAT by 5.9%. Esterification by ACAT and LCAT was reduced, respectively, by 25 and 11% by the presence of an additional methyl group on the side chain of cholesterol at the C-24 position. This data supports our hypothesis that the structure of the sterol substrate has a significant effect on its esterification by ACAT or LCAT. PMID- 19679307 TI - A mechanical model of the gating spring mechanism of stereocilia. AB - The stereocilium is the basic sensory unit of nature's mechanotransducers, which include the cochlear and vestibular organs. In noisy environments, stereocilia display high sensitivity to miniscule stimuli, effectively dealing with a situation that is a design challenge in micro systems. The gating spring hypothesis suggests that the mechanical stiffness of stereocilia bundle is softened by tip-link gating in combination with active bundle movement, contributing to the nonlinear amplification of miniscule stimuli. To demonstrate that the amplification is induced mechanically by the gating as hypothesized, we developed a biomimetic model of stereocilia and fabricated the model at the macro scale. The model consists of an inverted pendulum array with bistable buckled springs at its tips, which represent the mechanically gated ion channel. Model simulations showed that at the moment of gating, instantaneous stiffness softening generates an increase in response magnitude, which then sequentially occurs as the number of gating increases. This amplification mechanism appeared to be robust to the change of model parameters. Experimental data from the fabricated macro model also showed a significant increase in the open probability and pendulum deflection at the region having a smaller input magnitude. The results demonstrate that the nonlinear amplification of miniscule stimuli is mechanically produced by stiffness softening from channel gating. PMID- 19679308 TI - Can sulci protect the brain from traumatic injury? AB - The influence of sulci in dynamic finite element simulations of the human head has been investigated. First, a detailed 3D FE model was constructed based on an MRI scan of a human head. A second model with a smoothed brain surface was created based on the same MRI scan as the first FE model. These models were validated against experimental data to confirm their human-like dynamic responses during impact. The validated FE models were subjected to several acceleration impulses and the maximum principle strain and strain rate in the brain were analyzed. The results suggested that the inclusion of sulci should be considered for future FE head models as it alters the strain and strain distribution in an FE model. PMID- 19679309 TI - Bilateral knee osteoarthritis does not affect inter-joint coordination in older adults with gait deviations during obstacle-crossing. AB - Fifteen elderly subjects with bilateral medial knee osteoarthritis (OA) and 15 healthy elderly subjects walked and crossed obstacles with heights of 10%, 20%, and 30% of their leg lengths while sagittal angles and angular velocities of each joint were measured and their phase angles (phi) calculated. Continuous relative phase (CRP) were also obtained, i.e., phi(hip-knee) and phi(knee-ankle). The standard deviations of the CRP curve points were averaged to obtain deviation phase (DP) values for the stance and swing phases. Significant differences between the OA and control groups were found in several of the peak and crossing angles, and angular velocities at the knee and ankle. Both groups had similar CRP patterns, and the DP values of the hip-knee and knee-ankle CRP curves were not significantly different between the two groups. Despite significant changes in the joint kinematics, knee OA did not significantly change the way the motions of the lower limb joints are coordinated during obstacle-crossing. It appears that the OA groups adopted a particular biomechanical strategy among all possible strategies that can accommodate the OA-induced changes of the knee mechanics using unaltered inter-joint coordination control. This enabled the OA subjects to accommodate reliably the mechanical demands related to bilateral knee OA in the sagittal plane during obstacle-crossing. Maintaining normal and reliable inter joint coordination may be considered a goal of therapeutic intervention, and the patterns and variability of inter-joint coordination can be used for the evaluation of treatment effects. PMID- 19679310 TI - A study of the viability of obtaining a generic animation of the foot while walking for the virtual testing of footwear using dorsal pressures. AB - Establishing the appropriate pressure exerted by the shoe upper over the foot surface is fundamental for the design of specific footwear, although measuring the dorsal pressures can also provide important additional information. In previous works, a virtual simulator to perform studies of comfort and functionality in CAD footwear design was presented. This paper describes the procedure carried out to obtain the foot animations used in this simulator. The virtual feet used in the simulator are feet without a standard form scanned in a static way. Their movements are rebuilt from the register of movements of several foot anatomical points during a complete step. The dorsal pressures exerted by some shoe uppers on these anatomical points were measured for several subjects and used to establish the viability of the use of these animations in a virtual simulator for footwear. PMID- 19679311 TI - Synergetic microorganismic convection generated by Opercularia asymmetrica ciliates living in a colony as effective fluid transport on the micro-scale. AB - Ciliates, being one of the main substrates in granular activated sludge (GAS) formation, are treated as a major factor in granulation process. Cilia beats of Opercularia asymmetrica provide a continuous nutrient flux, enhancing the colonization of bacteria on Peritrichia stalks. Given that the ciliates tend to live in colonies, the main focus of the present work was an analysis and comparison of the flow effects induced by a single ciliate and by a colony. Investigations of the flow generated by Opercularia asymmetrica were carried out using micro-particle image velocimetry with biocompatible seeding. The results obtained showed different flow structures for a single ciliate and a colony. Moreover, the synergetic work of Opercularia asymmetrica living in a colony is considered as effective fluid transport. Additionally, analysis of the shear and normal strain rates provided information on mixing phenomena within the fluid on the micro-scale. The influence of seeding substance concentration on the flow pattern was also studied. PMID- 19679312 TI - Quantification of organic acids in particulate matter by coupling of thermally assisted hydrolysis and methylation with thermodesorption-gas chromatography-mass spectrometry. AB - A quantitative method for the determination of organic acids in atmospheric particles is developed. The method couples a derivatisation step (thermally assisted hydrolysis and methylation) and a Curie point pyrolyser as a thermal desorption technique and gas chromatography-mass spectrometry (CPP-GC-MS). Among the reagents tested (tetramethylammonium hydroxide (TMAH), tetramethylammonium acetate (TMAAc) and phenyltrimethylammonium hydroxide (TMPAH)), the best performance was found using TMAAc as a derivatisation reagent for the reaction time of 4s at 510 degrees C as heating temperature. Calibration was performed for a series of fatty acids (FA), dicarboxylic acids (DCA) and terpenoic acids (TA) under these conditions. Coefficients of determination (R(2)) were between 0.94 and 0.98. Limits of detection (LOD) were in the nanogram-range between 0.1 and 3.6 ng. The method is applied on atmospheric particle samples to obtain the quantification reproducibility and quantification limits. Reproducibility was determined in terms of relative standard deviations (RSD) for ambient aerosol samples collected by a high-volume-sampler (HVS, RSD=6-45%, n=10) and a Berner impactor (BI, RSD=5-34%, n=10). Based on 24h sampling time the developed method enables quantification of all three classes of acids for both sampling techniques. Calibration data and presented volume concentrations are compared with literature data. A comparison with an off-line methylation-GC-MS using BF(3) as a derivatisation reagent and capillary electrophoresis coupled mass spectrometry (CE-MS) showed a good agreement. Minimal sample preparation is the main advantage of the developed method. Depending on the sensitivity requirements the present method can be a fast and simple alternative to GC-MS techniques with conventional sample preparation steps for semi-volatile organic acids. PMID- 19679313 TI - Computational method for modeling of gradient separation in ion-exchange chromatography. AB - A model for the simulation of the gradient separation in ion-exchange chromatography is presented. It is based on discontinuous plate model and simulates the distribution of analytes in the ion-exchange column during the separation process. It enables calculations of chromatograms for the analytes with integer and non-integer effective charges under complex gradient profiles. Equilibrium concentrations of all analytes are calculated using the same mathematical equations and expressions regardless of the effective charge on the analyte. The main parameters required for the simulations have to be determined under isocratic elution. The suitability of the model was tested with different types of gradients. A comparison of retention times and chromatograms shows that reliable predictions for all tested gradients are achieved. The observed average of the absolute values of the relative errors of the retention times obtained for any analyte in the present study from the calculated chromatograms is below 4%, while the average error considering all analytes in the study is below 2%. PMID- 19679314 TI - Steady and dynamic rheological behaviors of sodium carboxymethyl cellulose entangled semi-dilute solution with opposite charged surfactant dodecyl trimethylammonium bromide. AB - The steady and dynamic rheological behaviors of sodium carboxymethyl cellulose (NaCMC) entangled semi-dilute solution filled with different concentrations of dodecyl-trimethylammonium bromide (C(12)TAB) were investigated. The results reveal that the zero shear rate viscosity (eta(0)) and dynamic modules (G'and G'') increase with C(12)TAB concentration (C(s)), and there exist three scaling regions divided by two critical C(12)TAB concentrations (C(1), C(2) and C(1)('),C(2)('), respectively, from steady and dynamic tests). The increase of viscosity and modules with C(s) is ascribed to formation of network due to C(12)TAB micelles bridging NaCMC chains. The two critical C(12)TAB concentrations implies that the structure evolution of NaCMC-C(12)TAB complex is exposed to three states with increasing C(s), i.e., no network formation, network extent progressive formation and perfect network formation, respectively. Moreover, C(1)('),C(2)(') are a little lower than C(1), C(2), indicating that the dynamic test is more sensitive to detect the structure change of the complex as compared with steady test. Furthermore, it is found that as NaCMC concentration increases, C(1)(C(1)(')),C(2)(C(2)(')), C(1)(C(1)('))-CAC and C(2)(C(2)('))-CAC increase. PMID- 19679315 TI - Dissipation mechanisms in ionic liquids. AB - The spreading of ionic liquids on molecularly smooth solid surfaces has been little studied in the past. We show that the spreading behaviors of the two ionic liquids, [EMIM] ethyl sulfate and ECOENG 500, are well described by the combined molecular kinetic and hydrodynamic model of de Ruijter, de Coninck, and Oshanin [M.J. de Ruijter, J. de Coninck, G. Oshanin, Langmuir 15 (1999) 2209] with reasonable values for the molecular friction coefficient zeta, molecular displacement lambda, and frequency K0 associated with contact line motion, as well as reasonable values for the microscopic cutoff a associated with hydrodynamic dissipation. PMID- 19679316 TI - Ratiometric pH-nanosensors based on rhodamine-doped silica nanoparticles functionalized with a naphthalimide derivative. AB - This paper describes the preparation of two-dye-doped silica nanoparticles for ratiometric pH measurements in the biologically relevant pH-range. While a rhodamine derivative is embedded in a silica core and used as the reference, a pH sensitive naphthalimide dye is immobilized on the previously amino-functionalized core through two different approaches. Either the naphthalimide's carboxylic group is activated to a succinimidyl-ester to form an amide bond or the system can be built up via solid-phase organic synthesis in only two steps. Both types of nanosensors are characterized in terms of morphology (dynamic light scattering, transmission electron microscopy) and optical properties (steady state fluorescence spectroscopy). In terms of application, e.g. reproducibility and handling of the synthesis, the first approach gave very good results with respect to size and size distribution and a pK(a) value of 6.55 was found that is comparable to the free indicator dye in solution. The solid-phase organic synthesis method proves the possibility of covalent immobilization of naphthalimides to amino-functionalized surfaces, showing the stability of the polymeric substrate and achieving comparable results for pH sensing. PMID- 19679317 TI - Phenol removal from aqueous solution by adsorption and ion exchange mechanisms onto polymeric resins. AB - The removal of phenol from aqueous solution was evaluated by using a nonfunctionalized hyper-cross-linked polymer Macronet MN200 and two ion exchange resins, Dowex XZ (strong anion exchange resin) and AuRIX 100 (weak anion exchange). Equilibrium experimental data were fitted to the Langmuir and Freundlich isotherms at different pHs. The Langmuir model describes successfully the phenol removal onto the three resins. The extent of the phenol adsorption was affected by the pH of the solution; thus, the nonfunctionalized resin reported the maximum loading adsorption under acidic conditions, where the molecular phenol form predominates. In contrast both ion exchange resins reported the maximum removal under alkaline conditions where the phenolate may be removed by a combined effect of both adsorption and ion exchange mechanisms. A theoretical model proposed in the literature was used to fit the experimental data and a double contribution was observed from the parameters obtained by the model. Kinetic experiments under different initial phenol concentrations and under the best pH conditions observed in the equilibrium experiments were performed. Two different models were used to define the controlling mechanism of the overall adsorption process: the homogeneous particle diffusion model and the shell progressive model fit the kinetic experimental data and determined the resin phase mechanism as the rate-limiting diffusion for the phenol removal. Resins charged after the kinetic experiments were further eluted by different methods. Desorption of nonfunctionalized resin was achieved by using the solution (50% v/v) of methanol/water with a recovery close to 90%. In the case of the ion exchange resins the desorption process was performed at different pHs and considering the effect of the competitive ion Cl-. The desorption processes were controlled by the ion exchange mechanism for Dowex XZ and AuRIX 100 resins; thus, no significant effect for the addition of Cl- under acidic conditions was observed, while under alkaline conditions the total recovery increased, specially for Dowex XZ resin. PMID- 19679318 TI - Cell transplantation in the treatment of acute liver injury: effect but no clear mechanism. PMID- 19679319 TI - Alkylphenols in the core sediment of a waste dumpsite in the East Sea (Sea of Japan), Korea. PMID- 19679320 TI - [Overestimation of the capillary glucose readings]. PMID- 19679321 TI - [Imported malaria in a county hospital: the new-old diseases]. PMID- 19679322 TI - [Assessment of nutritional quality in healthy pregnant women of the Canary Islands, Spain]. AB - BACKGROUND AND OBJECTIVE: To describe the composition of the diet of healthy pregnant women of the Canary Islands and to estimate the nutritional quality using the Healthy Eating Index (HEI). PATIENTS AND METHOD: Cross-sectional study based on 103 women aged 18-40 years, who gave birth at the University Hospital Materno-Infantil of Gran Canaria. Food consumption and macro and micronutrient intake were estimated using a food frequency questionnaire used in the Canary Island Nutrition Survey (ENCA) and the HEI was calculated. This index includes 10 components and the maximum possible score of the index is 100 points. RESULTS: The score of the index was 54.9. This result remains below the optimum score of > or =80, which is considered a diet of good quality of pregnant women in our study population. The average score of the first 5 components of the index showed that cereal consumption was below the daily portions recommended for pregnant women, whereas vegetables, fruit, milk and meat consumption surpassed the recommendations. A significant number of pregnant women did not reach the 50% of the recommendations for iron, folate and vitamin D intake (36.9, 26.2 and 38.8% respectively). At least 30% of the population exceeded 200% of the recommendations for proteins, thiamine, niacin, riboflavin, vitamin C and vitamin A. CONCLUSIONS: Dietary advice for improving the diet quality during pregnancy and the supplementation of mainly iron and folate are necessary. PMID- 19679323 TI - Hydroperoxy-cycloartane triterpenoids from the leaves of Markhamia lutea, a plant ingested by wild chimpanzees. AB - In the framework of the phytochemical investigation of plant species eaten by wild chimpanzees in their natural environment in Uganda, leaf samples of Markhamia lutea were selected and collected. The crude ethyl acetate extract of M. lutea leaves exhibited significant in vitro anti-parasitic activity and low cytotoxicity against MRC5 and KB cells. Fractionation of this extract led to six cycloartane triterpenoids, musambins A-C and their 3-O-xyloside derivatives musambiosides A-C. The structures were elucidated on the basis of spectral studies including mass spectroscopy and extensive 2D NMR. Most of the compounds exhibited mild anti-leishmanial and anti-trypanosomal activities. PMID- 19679324 TI - Cigarette-related injuries to young children in the USA, 2002-2007. PMID- 19679325 TI - The effect of low levels of dietary cobalt on the chemiluminescence response of polymorphonuclear leukocytes of goats. AB - Twenty ten-week-old newly weaned male Batinah goats were randomly assigned to a control (n=10) and a treated (n=10) group and were fed a diet containing 0.1mg/kg DM cobalt (Co). Goats in the treated group received bi-monthly subcutaneous injections of 2000 microg of hydroxycobalamin. The phagocytic function of the polymorphonuclear leukocytes (PMN) were tested using a luminol-dependent chemiluminescence assay with opsonized zymosan as the phagocytic target. One month after the onset of the experiment PMN from the control group exhibited a significantly (p<0.05) lower CL response, which continued for the second month. The results of the present study demonstrated that low levels of dietary cobalt leads to an early impairment of phagocytic function. This may at least in part, be an explanation as to why at the field level in Oman young goats fed diets containing low levels of Co appear to be more susceptible to infections. PMID- 19679326 TI - [Maxillary and mandibular carcinogenesis: research and prospects]. AB - Development and growth of odontogenic tumours depend on impairment of numerous genes and molecules. In recent years, most of the genes involved in dental development were identified. This produced a new basis for the study of oral pathology and maxillofacial carcinogenesis. A better understanding of these molecular phenomena should allow to better determine the evolution of such lesions. Research breakthroughs should facilitate the development of new molecular and genetic therapeutic perspectives. PMID- 19679328 TI - Diethanolamine Pd(II) complexes in bioorganic modeling as model systems of metallopeptidases and soybean lipoxygenase inhibitors. AB - The reaction of PdCl(2) with diethanolammonium chloride (DEAxHCl), in the molar ratio 1:2, affords the [HDEA](2)[PdCl(4)] complex (1). The hydrolytic activity of the novel Pd(II) complex 1 was tested in reaction with N-acetylated L histidylglycine dipeptide (AcHis-Gly). Complex 1, as well as earlier prepared trans-[PdCl(2)(DEA)(2)] complex (2), and DEA, as their precursor, were tested for their in vitro free radical scavenging activity. UV absorbance-based enzyme assays were done in order to evaluate their inhibitory activity of soybean lipoxygenase (LOX). Also, assays with superoxide anion radical were done. The scavenging activities of the complexes were measured and compared with those of their precursors and caffeic acid. Complex 2 exhibits the highest antioxidant activity and the highest inhibitory effect against the soybean LOX. PMID- 19679327 TI - Epilepsy surgery for insular lesions. AB - OBJECTIVE: Due to the proximity of eloquent areas of the brain, the surgical treatment of insular lesions causing refractory epilepsy is considered difficult. We report here on our experience in this field. METHODS: We identified 24 patients (age: 1-62 years, mean 27) who underwent epilepsy surgery for an insular lesion from the epilepsy surgery data bank. We analyzed the preoperative diagnostics, surgical strategy and postoperative follow-up (duration: 12-168 months, mean 37.5) for functional morbidity and seizure outcome. RESULTS: Eight patients had strictly insular lesions while, in 16 cases, the lesion extended into the frontal (n=3) or temporal (n=8) lobe, or was multilobar (n=5). Sixteen resections (66.7%) were right-sided. Six patients required invasive EEG with implanted electrodes, while three had the aid of intraoperative electrocorticography. In 12 patients, continuous electrophysiological monitoring was used intraoperatively (phase reversal, motor evoked potentials) and, in seven, neuronavigation. In seven patients, only subtotal resection of the insular lesion was possible due to involvement of eloquent areas, and two patients required repeat surgery to complete the resection. Thirteen patients had glial/glioneural tumours (WHO grades I-III), 11 from non-neoplastic lesions. Postoperatively, two patients (8.3%) had a transient neurological deficit (hemiparesis and dysphasia, respectively). One patient had permanent hemihypaesthesia, another had permanent deterioration of preexistent hemiparesis and two had hemianopia as calculated deficit (16.6% rate of mild permanent morbidity). According to the International League against Epilepsy (ILAE) classification, 15 patients were totally seizure-free (62.5%, ILAE 1) and 79.2% had a satisfactory seizure outcome (ILAE 1-3). CONCLUSION: In selected patients, an individually tailored lesionectomy of insular lesions can be performed, with acceptable safety, to provide a high rate of satisfactory seizure relief. Indeed, even subtotal resection can result in effective seizure control. PMID- 19679329 TI - Spatial analysis of human health risk associated with ingesting manganese in Huangxing Town, Middle China. AB - This paper spatially analyzed human health risk associated with ingesting manganese (Mn) contents in groundwater and vegetables irrigated with contaminated pond water in Huangxing Town, Middle China. The combination of monitoring data and sequential indicator simulation (SIS) was used to determine Mn exposure distributions in pond water and groundwater. Hazard quotient (HQ) associated with ingesting Mn was calculated to evaluate the risk to human health. Many HQs determined from risks exceed 1 in the region, indicating that the use of groundwater and pond water poses potential risk to human health. Lower risk areas are located in the northwest and partly southeast of the region. The probabilistic risk assessment formulated suitable references for pollution remedy and control in Huangxing Town. Safe areas in 75th percentile of HQ map are suggested to be safe for use and, the manganese residues in the unsafe areas of the 25th percentile of HQ map is to be treated firstly. PMID- 19679330 TI - Effect of humate on biological treatment of wastewater containing heavy metals. AB - This paper presents results of investigations on the influence of humic substances (humate, HS) on the biological treatment of wastewater containing heavy metals (Cr,Cu,Fe,Mn,Ni, and Zn). Respirometric studies indicated that the investigated system complied with the Haldane model for inhibitory wastes. Chemical analyses showed that, while the soluble COD removal was high (82%), only 7% of ammonia was oxidized to nitrate. An addition of HS (500mg L(-1)) mitigated the inhibitory effect of the wastewater on the returned activated sludge. The system with HS complied with the Monod model for non-inhibitory wastes, and the removal of ammonia and metals was 99% and over 90%, respectively. It is suggested that an application of HS could be beneficial for treatment plants receiving wastewater streams containing heavy metals. PMID- 19679331 TI - Removal of toxic ions (chromate, arsenate, and perchlorate) using reverse osmosis, nanofiltration, and ultrafiltration membranes. AB - Rejection characteristics of chromate, arsenate, and perchlorate were examined for one reverse osmosis (RO, LFC-1), two nanofiltration (NF, ESNA, and MX07), and one ultrafiltration (UF and GM) membranes that are commercially available. A bench-scale cross-flow flat-sheet filtration system was employed to determine the toxic ion rejection and the membrane flux. Both model and natural waters were used to prepare chromate, arsenate, and perchlorate solutions (approximately 100microgL(-1) for each anion) in mixtures in the presence of other salts (KCl, K(2)SO(4), and CaCl(2)); and at varying pH conditions (4, 6, 8, and 10) and solution conductivities (30, 60, and 115mSm(-1)). The rejection of target ions by the membranes increases with increasing solution pH due to the increasingly negative membrane charge with synthetic model waters. Cr(VI), As(V), and ClO(4)( ) rejection follows the order LFC-1 (>90%) > MX07 (25-95%) congruent withESNA (30 90%)>GM (3-47%) at all pH conditions. In contrast, the rejection of target ions by the membranes decreases with increasing solution conductivity due to the decreasingly negative membrane charge. Cr(VI), As(V), and ClO(4)(-) rejection follows the order CaCl(2)90%) excluding NO(3)(-) (71-74%) than the ESNA NF membrane (11-56%) with a relatively large pore size (0.44nm), indicating that size exclusion is at least partially responsible for the rejection. The ratio of solute radius (r(i,s)) to effective membrane pore radius (r(p)) was employed to compare ion rejection. For all of the ions, the rejection is higher than 70% when the r(i,s)/r(p) ratio is greater than 0.4 for the LFC-1 membrane, while for di-valent ions (CrO(4)(2-), SO(4)(2-), and HAsSO(4)(2-)) the rejection (38-56%) is fairly proportional to the r(i,s)/r(p) ratio (0.32-0.62) for the ESNA membrane. PMID- 19679332 TI - Fractionation of heavy metals in liquefied chromated copper arsenate 9-treated wood sludge using a modified BCR-sequential extraction procedure. AB - Chromated copper arsenate (CCA)-treated wood was liquefied with polyethylene glycol/glycerin and sulfuric acid. After liquefaction, most CCA metals (98% As, 92% Cr, and 83% Cu) were removed from liquefied CCA-treated wood by precipitation with calcium hydroxide. The original CCA-treated wood and liquefied CCA-treated wood sludge were fractionated by a modified Community Bureau of Reference (BCR) sequential extraction procedure. The purpose of the BCR-sequential extraction used in this study was to examine the availability of CCA metals in treated wood for reuse. Both As and Cr had a slightly higher concentration in the sludge sample than in original CCA-treated wood. The sequential extraction showed that As and Cr were principally existed in an oxidizable fraction (As, 67%; Cr, 88%) in original CCA-treated wood. Only 1% of both As and Cr were extracted by hot nitric acid with the last extraction step. The distribution of As and Cr changed markedly in liquefied CCA-treated wood sludge. The amount of As in the exchangeable/acid extractable fraction increased from 16% to 85% while the amount of Cr increased from 3% to 54%. Only about 3% of As was present in the oxidizable fraction. However, there was still about 34% of Cr in the same fraction. Based on these results from sequential extraction procedures, it can be concluded that the accessibilities of CCA metals increase markedly by the liquefaction-precipitation process. PMID- 19679333 TI - Monomorphism, male-male competition, and mechanisms of sexual dimorphism. PMID- 19679335 TI - Kidney internal splint/stent (KISS) catheter revisited for pediatric pyeloplasty. AB - OBJECTIVES: To review our experience using the Kidney Internal Splint/Stent (KISS) (Cook Medical, Bloomington, IN) to provide drainage after pyeloplasty. The KISS combines the attributes of nephrostomy tube diversion and anastomotic stent in a single tube. Since its initial description in 1993, additional reports on its use are limited. METHODS: We retrospectively reviewed charts of patients who underwent pyeloplasty by single surgeon, with simultaneous placement of KISS stent from 2003 to 2008. Preoperative and postoperative renal function and t((1/2)) times of the affected renal unit were determined by nuclear renography. Nephrostograms were performed 10 days after surgery. Complications from the use of KISS catheter including premature dislodgement or infection were noted. RESULTS: We performed 59 pyeloplasties with KISS stent placement in 57 patients. Mean age of patients was 23 months with follow-up of 32 months (range 6-69). Median preoperative and postoperative renal function was 47% and 49%, respectively. Median preoperative and postoperative t(1/2) times were 80 and 12 minutes, respectively (P = .001). Nephrostograms did not reveal leak in any patient. Unintentional removal of KISS stent did not occur in any patient. Postoperative febrile UTI occurred in 2 patients while the KISS stent was in place. CONCLUSIONS: KISS stents can be used safely and effectively in a wide variety of clinical situations surrounding pyeloplasty. The KISS catheter was well tolerated by the patients in our series. The KISS stent offers the combined advantages of nephrostomy tube and internal stent while obviating the second anesthetic that would be necessary with an internal stent. PMID- 19679336 TI - The case of a poorly differentiated gastric adenocarcinoma disguised as a testicular germ cell tumor in a 34-year-old male. PMID- 19679337 TI - Enlargement of accessory spleen after splenectomy can mimic a solitary adrenal tumor. AB - We report on a 72-year-old woman who had previously undergone splenectomy and subsequently presented with an incidental 5-cm adrenal mass. Laparoscopic adrenalectomy was performed, and the mass was identified to be an accessory spleen. Remaining accessory splenic tissue may undergo compensatory hypertrophy after splenectomy. When a biochemically inactive, well-marginated ovoid adrenal mass is identified in a postsplenectomy patient, consideration should be given to the presence of accessory spleen. In such cases, radionuclide imaging with technetium sulfur colloid may provide information that would confirm the presence of accessory normal tissue and would therefore support observation rather than surgical resection. PMID- 19679338 TI - Minimizing the cost of surgical correction of stress urinary incontinence and prolapse. AB - OBJECTIVES: To compare the operative costs associated with the use of incontinence kits, with or without biomaterials, with surgeon-tailored prolene mesh (STPM) in the treatment of stress urinary incontinence (SUI) with or without pelvic organ prolapse. METHODS: All operations for uncomplicated SUI with or without pelvic organ prolapse were reviewed from 2007-2008. Operative billing sheets including operative time, hospital cost, and the insurance billing statement were obtained and reviewed. Surgeon payment was not included in the analysis. Hospital stay was also compared. RESULTS: For patients with SUI alone, there was a significant difference in the hospital cost and the insurance billing statement between STPM and commercial kits (CK). On average, the insurance billing statement for STPM was $2220 less per case as compared with CK. For patients with SUI and anterior compartment prolapse, there was a significant difference in the hospital cost and the insurance billing statement between STPM and CK. On average, the insurance billing statement for STPM was $4770 less per case as compared with CK. For patients with SUI and anterior and posterior compartment prolapse, the difference in hospital cost and insurance billing statement approached statistical significance. The insurance billing statement for STPM on an average was $5600 less per case as compared with CK. There was no significant difference in operative time or hospital stay. CONCLUSIONS: The use of STPM for the treatment of incontinence with or without prolapse is significantly less costly for the hospital and the patient and/or insurance as compared with CK. The use of STPM did not increase operative time or postoperative hospital stay when compared with prefashioned kits. PMID- 19679339 TI - Prognostic relevance of prothymosin-alpha expression in human upper urinary tract transitional cell carcinoma. AB - OBJECTIVES: To investigate the prognostic role of prothymosin-alpha (PTMA) expression in human upper urinary tract transitional cell carcinoma (UUT-TCC). METHODS: Paraffin-embedded tissues were collected from 91 patients with UUT-TCC and from 15 paired normal renal cortex and 13 paired urothelial walls. The primary antibody for PTMA (2F11) used was validated in 4 human urothelial cancer cell lines before assessing the surgical specimen. Immunohistochemistry was then conducted to determine the expression intensity of PTMA, the calculation of immunostaining density using imaging analysis, and for immunostaining localization. The correlates with clinicopathologic characteristics and patient survival were explored. RESULTS: The expression intensity of PTMA demonstrated a significant enhancement of PTMA expression in UUT-TCCs compared with both paired normal tissues (P = .0002 and P = .0004 for UUT-TCC vs the urothelial wall and vs the renal cortex, respectively). As for the localization of PTMA immunoreactivity, of the 91 tumor specimens, 33 (36.3%) were cytoplasmic PTMA expressing, 51 (56.0%) were nuclear PTMA-expressing, and 7 (7.7%) were PTMA negative tumors. On univariate and multivariate analyses, PTMA expression localization was the sole independent prognostic indicator for recurrence-free survival (hazard ratio 4.90, 95% confidence interval 1.73-13.9; P = .003), although pathologic staging was an independent prognostic indicator for both progression-free survival (hazard ratio 22.6, 95% confidence interval 2.56-198; P = .005) and disease-specific overall survival (hazard ratio 5.60, 95% confidence interval 1.48-21.2; P = .011). The limitations of our study included small patient numbers and short follow-up. CONCLUSIONS: The results of our study have shown that PTMA is overexpressed in UUT-TCCs and that cytoplasmic PTMA expression can provide significant prognostic information for subsequent tumor recurrence in the residual urinary tract after nephroureterectomy. PMID- 19679340 TI - Retroperitoneoscopic nephrectomy for pyonephrotic nonfunctioning kidney. AB - OBJECTIVES: To review the feasibility and outcome of retroperitoneoscopic nephrectomy (RPN) for pyonephrotic nonfunctioning kidneys. METHODS: RPN for pyonephrotic nonfunctioning kidneys was used in 52 patients from July 2001 to May 2007. Percutaneous nephrostomy drainage was instituted in 46 patients because of sepsis before being referred to our institute. However, the remaining 6 patients underwent RPN without previous diversion. The mean patient age was 46.4 years (range 22-72). The etiology was stone disease in 29 patients, ureteropelvic junction obstruction in 18, and genitourinary tuberculosis in 5; 32 patients had diabetes mellitus as a comorbid condition. RESULTS: RPN was performed successfully in 46 patients (88.5%); 6 patients required conversion to open surgery (1 emergently because of colonic injury and 5 electively because of nonprogression of the procedure). In 6 patients, subcapsular nephrectomy was required. The mean operating time was 110 minutes (range 90-180). The mean blood loss was 95 mL (range 80-300), and the mean analgesic requirement was 150 mg (range 50-400) of tramadol. Five patients had Clavien grade I, 7 had grade II, and 2 had grade IIIb complications. One patient required blood transfusion. The mean hospital stay was 3.6 days (range 2-8), and the mean return to normal activity was 14.2 days (range 11-21). CONCLUSIONS: RPN, although challenging, is safe, reliable, and successful for treatment of pyonephrotic nonfunctioning kidneys. PMID- 19679341 TI - Adjuvant radiation therapy in stage III node-positive uterine cancer. AB - OBJECTIVES: To determine the association of adjuvant radiotherapy and outcomes of women with stage III node-positive uterine cancer. METHODS: All patients with surgically-staged stage III node-positive uterine cancer from the Surveillance Epidemiology and End Results database of the US National Cancer Institute from 1988 to 2001 were identified. Data were analyzed using Kaplan-Meier and logistic regression methods. RESULTS: Of 943 women, the median age was 64 years (range: 28 93). 82.1%, 8.6%, and 6.8% were White, Black, and Asian respectively. The median number of removed nodes was 11. 54.9% had a single positive node and 45.1% had 2 5 positive nodes. Endometrioid, papillary serous, sarcomas, and clear cell carcinomas comprised of 69.7%, 16.3%, 9.9%, and 4.1% of histologies, respectively. 67.3% of the women underwent adjuvant radiotherapy with a 5-year disease-specific survival of 67.9% compared to 53.4% in those without radiotherapy (p<0.001). Adjuvant radiotherapy improved the survival from 54.4% to 74.3% (p<0.001) in those with a single positive node and from 52.4% to 59.7% (p=0.089) in those with 2-5 positive nodes. On multivariate analysis, older age, non-endometrioid histology, and lack of adjuvant radiotherapy remained as significant independent prognostic factors for worsened survival. CONCLUSIONS: Our data suggest that adjuvant radiotherapy is associated with a significant survival benefit in women with single-positive node endometrioid uterine cancers. Prospective clinical trials are warranted to confirm these findings. PMID- 19679342 TI - Fatigue limits of enamel bonds with moist and dry techniques. AB - OBJECTIVE: Shear fatigue limit (SFL) testing, coupled with shear bond strength (SBS) measurements can provide valuable information regarding the ability of adhesive systems to bond to mineralized tooth structures. The clinical technique for enamel bonding with adhesive resins has shifted from bonding to a thoroughly dried acid conditioned surface to a moist surface to facilitate dentin bonding. The purpose of this study was to compare the performance of ethanol-containing etch-and-rinse adhesive (ERA) systems on moist and dry enamel by determining the resin composite to enamel SBS and SFL, and examining the relationship of SBS and SFL. METHODS: Twelve specimens each were used to determine 24-h resin composite (Z100 - 3M ESPE) to enamel SBS to moist and dry surfaces with two ERA systems, Adper Single Bond Plus (SBP) and OptiBond Solo Plus (OBP). A staircase method of fatigue testing was used in a four-station fatigue cycler to determine the SFL of resin composite to enamel bonds (moist and dry) with the two ERA systems (20 specimens for each test condition) at 0.25Hz for 40,000 cycles. ANOVA and Tukey's post hoc test were used for the SBS data and a modified t-test with Bonferroni correction was used for comparisons of SFL. RESULTS: The two ERA systems each generated statistically similar SBS (p>0.05) to moist and dry enamel and the SBS of SBP was significantly higher than OBP on dry enamel (p<0.05). The SFL of SBP was significantly greater to dry enamel when compared to moist enamel and there was not a significant difference in the SFL of OBP on dry and moist enamel. There were no significant differences in SFL values between SBP on either moist or dry enamel and OBP on both moist and dry enamel. SIGNIFICANCE: Fatigue testing may provide more useful information than SBS tests regarding the performance of dental adhesive systems. The chemical composition, solvents and filler components of ERA systems may influence their ability to develop long-term durable bonds to both moist and dry enamel surfaces. PMID- 19679344 TI - Public stewardship of mixed health systems. PMID- 19679343 TI - N-Acetyl cysteine (NAC) inhibits proliferation, collagen gene transcription, and redox stress in rat palatal mucosal cells. AB - OBJECTIVES: Control of hyperplastic and invasively growing gingival tissue is crucial for maintaining normal oral function and for successful bone regenerative therapy. We tested the hypothesis that materials containing N-acetyl cysteine (NAC), an antioxidant cysteine derivative, can control proliferation and function of oral mucosal cells. METHODS: Oral mucosal cells derived from the rat palatal tissue were cultured with or without NAC at different concentrations (2.5 10.0mM). To simulate inflammatory conditions, cultures were treated with hydrogen peroxide. NAC was also applied via collagen materials in membrane and sponge forms to explore the clinical applicability. The redox balance inside the cells was evaluated by measuring the concentration of intracellular glutathione (GSH). RESULTS: Adding NAC into cultures of oral mucosal cells reduced their proliferation, transcriptional expression, and collagen production in an NAC concentration-dependent manner without cytotoxic effects. Furthermore, NAC substantially reduced the hydrogen peroxide-induced elevation of cellular proliferation and collagen production. The controlling effects of NAC were also demonstrated in cells cultured on NAC-containing collagen materials and were associated with an increase in intracellular glutathione (GSH) reserves and a decrease in the oxidized form of glutathione (GSSG). SIGNIFICANCE: These results indicate that NAC may abrogate inflammation- or oxidative-stress-induced hyperfunction of oral mucosal cells and that it can be delivered effectively via biodegradable materials. This study provides a basis to explore NAC-containing biomaterials that are functionalized to control oral soft tissue growth and function without cytotoxicity. PMID- 19679346 TI - The ecological status of Karavasta Lagoon (Albania): Closing the stable door before the horse has bolted? AB - Karavasta is the widest and most important lagoon in Albania. This study aimed to assess the ecological quality status of the lagoon, acquire knowledge of a natural environment which might be exploited for aquaculture, and give management hints on the basis of anthropogenic impact and ecological conditions. A sampling campaign was carried out in 2008: at six stations, benthic fauna, water, and sediment parameters were considered. Statistical analyses were carried out through multivariate procedures (PCA, classification-clustering, SIMPER, RDA, DISTLM, PERMANOVA). Ecological quality was assessed through the AZTI Marine Biotic Index (AMBI), the multivariate AMBI (M-AMBI) and the Benthic Index based on Taxonomic Sufficiency (BITS). Sediment characteristics (percent organic matter, %OM; redox potential discontinuity layer depth, RPDL; particle size composition) and salinity represented contributory influences on lagoon communities. It was possible to distinguish and characterise a confined area, and benthic communities, from a marine-influenced area and its biota. The number of species was quite low when compared with other open Adriatic lagoons. The M-AMBI and BITS classifications gave quite similar results, which seemed consistent with the ecological conditions of the lagoon, that is a distinction in the ecological quality between the seaward and landward stations, with higher ecological quality (EcoQ) at the seaward stations. Given the pressures and the ecological condition of Karavasta, an intensification of aquaculture activities must be considered with caution, since the lagoon seems at significant risk of serious hypereutrophication. This situation is made worse by the limited water exchange with the marine environment due to the irregular dredging of the communication channels. PMID- 19679345 TI - How to maintain surveillance for novel influenza A H1N1 when there are too many cases to count. PMID- 19679347 TI - Activation of beta-catenin signalling increases StarD7 gene expression in JEG-3 cells. AB - StarD7 gene encodes a protein that belongs to the StAR-related lipid transfer proteins involved in intracellular transport and metabolism of lipids. It has been previously documented that StarD7 has a wide-spread mRNA expression in trophoblastic tissues and several tumour cell lines with highest levels in both choriocarcinoma JEG-3 and JAR cells, hepatocellular carcinoma HepG2, and colorectal adenocarcinoma HT-29 cells. To understand the molecular mechanisms that regulate the expression of the human StarD7 gene, we have cloned and characterized the 5'-flanking region of the gene. Transient transfections of several 5'deleted StarD7-promoter-firefly luciferase constructs into JEG-3 cells indicated that the -312/+157 region contains the gene minimal promoter. In addition, sequence analysis of a 1.6kb gene fragment revealed the presence of a TATA-less promoter as well as multiple regulatory motifs, including one regulatory element corresponding to the T-cell factor 4 (TCF4) binding site. Inhibition of glycogen synthase kinase-3beta (GSK3beta), a component of Wnt/beta catenin signalling, increased both StarD7 mRNA and protein expression as well as its promoter activity. Co-transfection experiments in JEG-3 cell line revealed that the StarD7 promoter is activated by TCF4 transcription factor and by its beta-catenin coactivator. Moreover, site-directed mutagenesis of the TCF4 site located -614/-608bp relative to the transcription start site markedly diminished StarD7 promoter activity. Chromatin immunoprecipitation analysis demonstrated that beta-catenin and TCF4 are bound in vivo to the StarD7 gene promoter in JEG-3 cells treated with lithium chloride. Collectively, these studies show that beta catenin and TCF4 activate the human StarD7 gene interacting with its promoter region through Wnt/beta-catenin signalling. PMID- 19679348 TI - Unearthing the roles of imprinted genes in the placenta. AB - Mammalian fetal survival and growth are dependent on a well-established and functional placenta. Although transient, the placenta is the first organ to be formed during pregnancy and is responsible for important functions during development, such as the control of metabolism and fetal nutrition, gas and metabolite exchange, and endocrine control. Epigenetic marks and gene expression patterns in early development play an essential role in embryo and fetal development. Specifically, the epigenetic phenomenon known as genomic imprinting, represented by the non-equivalence of the paternal and maternal genome, may be one of the most important regulatory pathways involved in the development and function of the placenta in eutherian mammals. A lack of pattern or an imprecise pattern of genomic imprinting can lead to either embryonic losses or a disruption in fetal and placental development. Genetically modified animals present a powerful approach for revealing the interplay between gene expression and placental function in vivo and allow a single gene disruption to be analyzed, particularly focusing on its role in placenta function. In this paper, we review the recent transgenic strategies that have been successfully created in order to provide a better understanding of the epigenetic patterns of the placenta, with a special focus on imprinted genes. We summarize a number of phenotypes derived from the genetic manipulation of imprinted genes and other epigenetic modulators in an attempt to demonstrate that gene-targeting studies have contributed considerably to the knowledge of placentation and conceptus development. PMID- 19679349 TI - The presence of kinesin superfamily motor proteins KIFC1 and KIF17 in normal and pathological human placenta. AB - Kinesin superfamily proteins (KIFs) are motor proteins that participate in chromosomal and spindle movements during mitosis and meiosis, and transport membranous organelles and macromolecules fundamental for cellular functions. Although the roles of KIFs in axonal and dendritic transports have been studied extensively, their role in intracellular transport in general is less well known. The diversity of kinesins suggests that each kinesin may have a specific function. Therefore, in this study we aimed to investigate the presence and cellular localization of KIFC1 and KIF17 in normal and pathological human placentas. First-trimester (22-56 days) and normal, preeclamptic (PE), and diabetic-term placental tissues were obtained and further studied by immunohistochemistry (IHC) and Western blot methods. KIFC1 was mainly localized to the syncytiotrophoblast both in early and term placental samples. However, a stronger immunoreactivity was observed both in PE and diabetic placentas compared to normal-term placentas. KIF17 was most intensively localized in developing vascular endothelium in early pregnancy. Even though KIF17 was moderately stained in the endothelium of villi from normal human-term placentas, stronger immunoreactivity was observed in all types of villi of both PE and diabetic placentas. Western blotting of tissue extracts confirmed the IHC results. Here, we demonstrate the presence of KIFC1 and KIF17 in human placenta for the first time. The intense expression of KIFC1 in syncytiotrophoblast and KIF17 in vascular endothelium suggests that both the proteins might be important in a cargo-transport system. An increased expression pattern of both KIFC1 and KIF17 in PE and diabetes might suggest that these proteins may be involved in complex trophoblast functions and placental pathologies. Further studies will clarify the physiological role of KIFs in human placental transport and development. PMID- 19679350 TI - Variable luminal sarcoplasmic reticulum Ca(2+) buffer capacity in smooth muscle cells. AB - Sarcoplasmic reticulum contains the internal Ca(2+) store in smooth muscle cells and its lumen appears to be a continuum that lacks diffusion barriers. Accordingly, the free luminal Ca(2+) level is the same all throughout the SR; however, whether the Ca(2+) buffer capacity is the same in all the SR is unknown. We have estimated indirectly the luminal Ca(2+) buffer capacity of the SR by comparing the reduction in SR Ca(2+) levels with the corresponding increase in [Ca(2+)](i) during activation of either IP(3)Rs with carbachol or RyRs with caffeine, in smooth muscle cells from guinea pig urinary bladder. We have determined that carbachol-sensitive SR has a 2.4 times larger Ca(2+) buffer capacity than caffeine-sensitive SR. Rapid inhibition of SERCA pumps with thapsigargin revealed that this pump activity accounts for 80% and 60% of the Ca(2+) buffer capacities of carbachol- and caffeine-sensitive SR, respectively. Moreover, the Ca(2+) buffer capacity of carbachol-sensitive SR was similar to caffeine-sensitive SR when SERCA pumps were inhibited. Similar rates of Ca(2+) replenishments suggest similar levels of SERCA pump activities for either carbachol- or caffeine-sensitive SR. Paired pulses of caffeine, in conditions of low Ca(2+) influx, indicate the relevance of luminal SR Ca(2+) buffer capacity in the [Ca(2+)](i) response. To further study the importance of luminal SR Ca(2+) buffer capacity in the release process we used low levels of heparin to partially inhibit IP(3)Rs. This condition revealed carbachol-induced transient increase of luminal SR Ca(2+) levels provided that SERCA pumps were active. It thus appears that SERCA pump activity keeps the luminal SR Ca(2+)-binding proteins in the high capacity, low-affinity conformation, particularly for IP(3)R-mediated Ca(2+) release. PMID- 19679352 TI - Primary hepatic MALT lymphoma associated with primary biliary cirrhosis. PMID- 19679351 TI - Circulating Ki-67 protein in plasma as a biomarker and prognostic indicator of acute lymphoblastic leukemia. AB - Tissue-based determination of Ki-67, a marker of cellular proliferation, has shown prognostic value in solid tumors and hematological malignancies. We developed and validated an electrochemiluminescence-based method for sensitive measurement of circulating Ki-67 in plasma (cKi-67). This assay demonstrated significantly higher levels of cKi-67 in patients with newly diagnosed acute lymphoblastic leukemia (ALL) (n=27; median, 762; range, 0-4574U/100 microL) than in healthy control subjects (n=114; median, 399; range, 36-2830U/100 microL). Moreover, elevated plasma cKi-67 was associated with significantly shorter survival in ALL patients (P=0.05). These findings suggest that Ki-67 can be detected in circulation and has potential for use as a biomarker for predicting clinical behavior in ALL. PMID- 19679353 TI - Constitutional RUNX1 deletion presenting as non-syndromic thrombocytopenia with myelodysplasia: 21q22 ITSN1 as a candidate gene in mental retardation. PMID- 19679354 TI - Erdheim-Chester disease: multisystem involvement and management with interferon alpha. PMID- 19679355 TI - An international meta-analysis of values of travel time savings. AB - Values of travel time savings are often used in cost-benefit analysis of transport projects and policies, and also to compute generalised travel costs. There has been considerable debate as to whether different research methods (e.g. stated versus revealed preference) will lead to different values of travel time savings, and which segmentations (e.g. by income or mode) are most important to capture the heterogeneity in these values. In addition there are many countries where no specific valuation studies have been done. In this paper new equations are estimated on the outcomes of value of travel time savings studies from various countries. In the data set, several countries appear more than once, which is taken into account by estimating random effects panel models. The meta analysis sheds some new light on the variation of the value of travel time savings by income, country, travel purpose, mode, distance and by survey method. Furthermore, the resulting meta-models are applied to produce new values of travel time savings for business travel, commuting and for other purposes in passenger transport, for 25 European Union Member states. Similar methods could be used to statistically analyse studies carried out on other non-monetary effects, both for transport and non-transport projects, and for inclusion in cost benefit analysis. PMID- 19679356 TI - Atypical fungal granuloma of the sphenoid wing. AB - A 29-year-old immunocompetent patient presented with a 3-month history of headache and vomiting. Computed tomography (CT) and conventional magnetic resonance imaging (MRI) revealed a mass lesion in the right sphenoid wing. The conventional imaging findings were typical of meningioma. However, diffusion weighted imaging (DWI), susceptibility-weighted imaging (SWI) and perfusion weighted imaging (PWI) all revealed details that were unusual for a meningioma. DWI showed diffusion blackout, perfusion was not raised in PWI, and susceptibility effects were noted in SWI. Based on these findings, the possibility of granuloma was kept as the differential diagnosis. Histopathological examination of the lesion was suggestive of fungal granuloma. This case report highlights the importance of advanced neuroimaging techniques in differentiating meningioma and granuloma. PMID- 19679357 TI - Superoxide protects Escherichia coli from bleomycin mediated lethality. AB - Superoxide and its products, especially hydroxyl radical, were recently proposed to be instrumental in cell death following treatment with a wide range of antimicrobials. Surprisingly, bleomycin lethality to Escherichia coli was ameliorated by a genetic deficiency of superoxide dismutase or by furnishing the superoxide generator plumbagin. Rescue by plumbagin was similar in strains containing or lacking recA or with inactive, inducible, or constitutive soxRS regulons. Thus, superoxide interferes with bleomycin cytotoxicity in ways not readily explained by genetic pathways expected to protect from oxidative damage. PMID- 19679358 TI - Brief self-rated screening for depression on the Internet. AB - BACKGROUND: The Internet offers promising possibilities for the quick screening of depression for treatment and research purposes. This paper aims to validate three self-rated measures to screen for depression on the Internet: SID (single item depression scale), CES-D (Center for Epidemiological Studies Depression scale) and K10 (Kessler psychological distress scale). METHODS: Of the 502 subjects aged 18-80 who rated the SID, CES-D and K10 measures on the Internet, 157 (31%) subjects were also interviewed by telephone using the WHO Composite International Diagnostic Interview (C)IDI) for DSM-IV-disorders. RESULTS: Cronbach's alpha for both web self-rated measures CES-D and K10 was 0.90. The SID correlated 0.68 (P<0.001) with the CES-D and with the K10. The CES-D correlated 0.84 with the K10 (P<0.001). Subjects with a DSM-IV diagnosis for any depressive disorder had significantly higher means (P<0.001) on the three self-rated measures for depressive symptoms than subjects without a diagnosis of any depressive disorder. Using any depressive disorder as the gold standard, the area under the curve (AUC) of the SID was 0.71 (95% CI: 0.63-0.79), which was significantly lower than the AUC of the CES-D (AUC: 0.84; 95% CI: 0.77-0.90, P=0.003) and of the K10 (AUC: 0.81; 95% CI: 0.73-0.88, P=0.0024). The AUCs for the K10 and CES-D did not differ significantly from each other. LIMITATIONS: The CIDI interviews were not recorded, so inter-rater reliability could not be calculated. CONCLUSIONS: The CES-D and K10 are reliable, valid tools for care providers to quickly screen depressive patients on the Internet and for researchers to collect data. PMID- 19679359 TI - Duration of sexual relationship and its effect on preeclampsia and small for gestational age perinatal outcome. AB - The aim of this study was to determine if women with preeclampsia or delivering small for gestational age (SGA) babies are more likely to have a short duration of sexual relationship compared with those who have uncomplicated pregnancies. In a prospective cohort study, 2507 nulliparous women with singleton pregnancies were interviewed at 15+/-1 weeks gestation about the duration of their sexual relationship with the biological father. Short duration of sexual relationship (< or =6 months, < or =3 months, or first intercourse) was compared between women with preeclampsia (N=131) or SGA babies (N=263) and those with uncomplicated pregnancies (N=1462). Short duration of sexual relationship was more common in women with preeclampsia compared with uncomplicated pregnancies (< or =6 months 14.5% versus 6.9%, adjusted odds ratio [adjOR] 1.88, 95% CI 1.05-3.36; < or =3 months 6.9% versus 2.5%, adjOR 2.32, 95% CI 1.03-5.25; first intercourse 1.5% versus 0.5%, adjOR 5.75, 95% CI 1.13-29.3). Although the total number of semen exposures was lower in SGA, SGA was not associated with a shorter duration of sexual relationship. On post hoc analysis, the subgroup of SGA with abnormal uterine artery Doppler at 20 weeks (N=58) were more likely to have had a short sexual relationship compared with controls (< or =6 months adjOR 2.33, 95% CI 1.09-4.98; < or =3 months adjOR 3.22, 95% CI 1.18-8.79; first intercourse adjOR 8.02, 95% CI 1.58-40.7). We conclude that compared to uncomplicated pregnancies, short duration of sexual relationship is more common in women who develop preeclampsia and women with abnormal uterine artery Doppler waveforms who deliver an SGA baby. PMID- 19679360 TI - Cell proliferation alterations in Chlorella cells under stress conditions. AB - Very little is known about growth and proliferation in relation to the cell cycle regulation of algae. The lack of knowledge is even greater when referring to the potential toxic effects of pollutants on microalgal cell division. To assess the effect of terbutryn, a triazine herbicide, on the proliferation of the freshwater microalga Chlorella vulgaris three flow cytometric approaches were used: (1) in vivo cell division using 5-,6-carboxyfluorescein diacetate succinimidyl ester (CFSE) staining was measured, (2) the growth kinetics were determined by cytometric cell counting and (3) cell viability was evaluated with the membrane impermeable double-stranded nucleic acid stain propidium iodide (PI). The results obtained in the growth kinetics study using CFSE to identify the microalgal cell progeny were consistent with those determined by cytometric cell counting. In all C. vulgaris cultures, each mother cell had undergone only one round of division through the 96 h of assay and the cell division occurred during the dark period. Cell division of the cultures exposed to the herbicide was asynchronous. Terbutryn altered the normal number of daughter cells (4 autospores) obtained from each mother cell. The number was only two in the cultures treated with 250 nM. The duration of the lag phase after the exposure to terbutryn could be dependent on the existence of a critical cell size to activate cytoplasmic division. Cell size, complexity and fluorescence of chlorophyll a of the microalgal cells presented a marked light/dark (day/night) cycle, except in the non-dividing 500 nM cultures, where terbutryn arrested cell division at the beginning of the cycle. Viability results showed that terbutryn has an algastatic effect in C. vulgaris cells at this concentration. The rapid and precise determination of cell proliferation by CFSE staining has allowed us to develop a model for assessing both the cell cycle of C. vulgaris and the in vivo effects of pollutants on growth and reproduction at microalgal cell level. PMID- 19679361 TI - A network against failing hearts--introducing the German "Competence Network Heart Failure". AB - Heart failure (HF) has been identified as one of the most threatening diseases for the western civilisation, posing a risk to health for a rising number of patients. Acknowledging the medical problem of HF to be both economically and socially threatening the German Federal Ministry of Research and Education (BMBF) initiated a nationwide research network aiming to find new ways in prevention, alleviation and treatment of the widespread disease. The "Competence Network Heart Failure" (CNHF), initiated in 2003, bundles the scientific expertise in a large-scale research network; its aims are the coordination of basic and applied clinical research as well as dissemination of findings into clinical practice in order to consolidate and perpetuate the achieved improvements. The scope of this paper is to introduce the CNHF and to provide an overview of the tasks and hitherto attained achievements to a broad spectrum of health care providers. PMID- 19679362 TI - Next-generation sequencing technologies and their implications for crop genetics and breeding. AB - Using next-generation sequencing technologies it is possible to resequence entire plant genomes or sample entire transcriptomes more efficiently and economically and in greater depth than ever before. Rather than sequencing individual genomes, we envision the sequencing of hundreds or even thousands of related genomes to sample genetic diversity within and between germplasm pools. Identification and tracking of genetic variation are now so efficient and precise that thousands of variants can be tracked within large populations. In this review, we outline some important areas such as the large-scale development of molecular markers for linkage mapping, association mapping, wide crosses and alien introgression, epigenetic modifications, transcript profiling, population genetics and de novo genome/organellar genome assembly for which these technologies are expected to advance crop genetics and breeding, leading to crop improvement. PMID- 19679363 TI - Dynamic template-assisted strategies in fragment-based drug discovery. AB - Fragment-based methods for drug discovery are increasingly popular because they provide drug leads with greater ligand efficiency than conventional high throughput screening. However, established methods for fragment detection do not address the central question in fragment-based ligand discovery: how can a primary ligand be optimally extended by a secondary fragment? Dynamic screening methods solve this issue by using a protein target as a template for ligand assembly, thus yielding high-affinity binders from low-affinity fragments. This review summarizes recent work on dynamic screening methodology, which resulted in the development of several high-affinity binders for various targets. Strengths and limitations of the published approaches are discussed and possible contributions of dynamic screening methodology to the drug discovery process are highlighted. PMID- 19679365 TI - Dose-effect relationship for local control of cervical cancer by magnetic resonance image-guided brachytherapy. AB - BACKGROUND AND PURPOSE: To analyse dose-response relationships for local control of cervical cancer after MR image-guided brachytherapy (IGBT) based on dose volume histogram parameters. METHODS AND MATERIALS: The analysis includes 141 patients with cervix cancer (stages IB-IVA) treated with 45-50.4 Gy EBRT+/ cisplatin plus 4 x 7 Gy IGBT. Gross tumour volume (GTV), high risk clinical target volume (HR CTV) and intermediate risk CTV (IR CTV) were delineated and DVH parameters (D90, D100) were assessed. Doses were converted to the equivalent dose in 2 Gy (EQD2) using linear-quadratic model (alpha/beta=10 Gy). Groups of patients were formed according to tumour size at diagnosis (GTV(D)) of 2-5 cm (group 1) or >5 cm (2), with subgroups of the latter for HR CTV size at first IGBT 2-5 cm (2a) or >5 cm (2b). Dose-response dependence for local recurrence was evaluated by logit analysis. RESULTS: Eighteen local recurrences in the true pelvis were observed. Dose-response analyses revealed a significant effect of HR CTV D100 (p=0.02) and D90 (p=0.005). The ED50-values for tumour control were 33+/ 15 Gy (D100) and 45+/-19 Gy (D90). ED90-values were 67 Gy (95% confidence interval [50;104]) and 86 Gy [77;113], respectively. CONCLUSIONS: A significant dependence of local control on D100 and D90 for HR CTV was found. Tumour control rates of >90% can be expected at doses >67 Gy and 86 Gy, respectively. PMID- 19679364 TI - Enhancement of the surface expression of G protein-coupled receptors. AB - G protein-coupled receptors (GPCRs) mediate physiological responses to a diverse array of stimuli and are the molecular targets for numerous therapeutic drugs. GPCRs primarily signal from the plasma membrane, but when expressed in heterologous cells many GPCRs exhibit poor trafficking to the cell surface. Multiple approaches have been taken to enhance GPCR surface expression in heterologous cells, including addition/deletion of receptor sequences, co expression with interacting proteins, and treatment with pharmacological chaperones. In addition to providing enhanced surface expression of certain GPCRs in heterologous cells, these approaches have also shed light on the control of GPCR trafficking in vivo and in some cases have led to new therapeutic approaches for treating human diseases that result from defects in GPCR trafficking. PMID- 19679366 TI - Weekly paclitaxel with concurrent radiotherapy followed by adjuvant chemotherapy in locally advanced nasopharyngeal carcinoma. AB - PURPOSE: To evaluate the efficacy and toxicity of weekly paclitaxel with concurrent radiotherapy followed by adjuvant chemotherapy (AC) in patients with locally advanced nasopharyngeal carcinoma (NPC). METHODS AND MATERIALS: Between 2004 and 2007, 54 patients with locally advanced NPC were included in this protocol. PATIENT CHARACTERISTICS: median age 48; 69% male; 52% World Health Organization (WHO) III; 50% stage III, 50% stage IV. The patients underwent a course of definitive conventional radiotherapy (70 Gy in 7 weeks with 2 Gy/fraction), with concurrent weekly paclitaxel 35 mg/m(2) from the first to the sixth week of radiation. AC was started 4 weeks after the end of the radiotherapy (RT), paclitaxel 135 mg/m(2) on day 1 and cisplatin 30 mg/m(2) on days 1-3 were administered every 4 weeks for two cycles. RESULTS: Median follow-up was 32 months. Eighty-five percentage of complete response and 15% partial response were achieved at the time of one month after AC. The 3-year actuarial rate of local regional control was 86%; distant metastases-free survival, progression-free survival and overall survival at 3 years were 81%, 69% and 76%, respectively. Forty-nine (91%) patients completed six courses of concurrent chemotherapy with weekly paclitaxel, and 4 (7%) patients delayed at the second cycle of AC. No patient developed severe acute toxicities. CONCLUSIONS: Weekly paclitaxel with concurrent RT followed by AC is a potentially effective and toxicity tolerable method for locally advanced NPC. Further studies are needed to identify the optimal dose of weekly paclitaxel in this strategy. PMID- 19679367 TI - Biological risks associated with consumption of reptile products. AB - The consumption of a wide variety of species of reptiles caught from the wild has been an important source of protein for humans world-wide for millennia. Terrapins, snakes, lizards, crocodiles and iguanas are now farmed and the consumption and trade of their meat and other edible products have recently increased in some areas of the world. Biological risks associated with the consumption of products from both farmed and wild reptile meat and eggs include infections caused by bacteria (Salmonella spp., Vibrio spp.), parasites (Spirometra, Trichinella, Gnathostoma, pentastomids), as well as intoxications by biotoxins. For crocodiles, Salmonella spp. constitute a significant public health risk due to the high intestinal carrier rate which is reflected in an equally high contamination rate in their fresh and frozen meat. There is a lack of information about the presence of Salmonella spp. in meat from other edible reptilians, though captive reptiles used as pets (lizards or turtles) are frequently carriers of these bacteria in Europe. Parasitic protozoa in reptiles represent a negligible risk for public health compared to parasitic metazoans, of which trichinellosis, pentastomiasis, gnathostomiasis and sparganosis can be acquired through consumption of contaminated crocodile, monitor lizard, turtle and snake meat, respectively. Other reptiles, although found to harbour the above parasites, have not been implicated with their transmission to humans. Freezing treatment inactivates Spirometra and Trichinella in crocodile meat, while the effectiveness of freezing of other reptilian meat is unknown. Biotoxins that accumulate in the flesh of sea turtles may cause chelonitoxism, a type of food poisoning with a high mortality rate in humans. Infections by fungi, including yeasts, and viruses widely occur in reptiles but have not been linked to a human health risk through the contamination of their meat. Currently there are no indications that natural transmissible spongiform encephalopathies (TSEs) occur in reptilians. The feeding of farmed reptiles with non-processed and recycled animal products is likely to increase the occurrence of biological hazards in reptile meat. Application of GHP, GMP and HACCP procedures, respectively at farm and slaughterhouse level, is crucial for controlling the hazards. PMID- 19679368 TI - Excellent effect of three-dimensional culture condition on pancreatic islets. AB - AIM: Culture of cells in simulated microgravity may be potentially beneficial to the fields of cell biology and somatic cell therapy. We aimed to examine three dimensional culture condition on pancreatic islets. METHODS: Islets of Langerhans were cultured in conditions of stasis, microgravity, and microgravity with a polyglycolic acid (PGA) fibrous scaffold. After 5 days in culture, islets were transplanted into the leg muscles of streptozotocin-treated diabetic Wistar rats. The blood glucose and insulin content were determined from the tail vein blood of recipients. The grafts were then frozen, dried, and coated for analysis by scanning electron microscopy. RESULTS: Grafts cultured in the three-dimensional conditions (simulated microgravity in the presence or absence of a PGA fibrous scaffold) were capable of significantly normalizing insulin production and blood glucose concentration when compared to control grafts (p<0.017). Scanning electron microscopy showed that the transplanted islets from three-dimensional culture groups demonstrated normal morphology with extracellular matrix on the surface. Islets in the PGA group exhibited well adhesion to PGA scaffolds. CONCLUSIONS: The three-dimensional culture conditions significantly improved the function and morphology of the grafts. The function and morphology of the grafts in the microgravity with a scaffold group was the excellent one. PMID- 19679369 TI - Effect of tight control of HbA1c and blood pressure on cardiovascular diseases in type 2 diabetes: an observational study from the Swedish National Diabetes Register (NDR). AB - AIM: To estimate hazard ratio (HR) of first incident fatal/non-fatal cardiovascular diseases (CVD) in female/male type 2 diabetic patients, with tight versus adverse control of HbA1c and blood pressure (BP) at baseline, age 30-70 years, no baseline CVD, followed for mean 5.7 years. METHODS: 2593 patients with tight control of HbA1c <7.5% and BP < or = 140/90 mmHg (median 6.5%/130/80 mmHg), and 2160 patients with adverse control 7.5-9.0%/141-190/91-110 mmHg (median 8.1%/155/85 mmHg). RESULTS: The hazard ratio (HR) for CVD with tight/adverse control was 0.67 (0.55-0.80; p<0.001), adjusting for age, sex, duration, hypoglycaemic treatment, smoking, BMI, lipid-lowering drugs, antihypertensive drugs, microalbuminuria. Adjusted HR for myocardial infarction, coronary heart disease, stroke and total mortality were 0.72 (0.56-0.92; p=0.01), 0.69 (0.55 0.86; p<0.001), 0.62 (0.45-0.84; p<0.001), 1.00 (0.72-1.39). The partial population-attributable risk percent for myocardial infarction, stroke and CVD was 23%, 33%, 29% if adverse HbA1c/BP control could be avoided, while 43%, 38%, 39% with overweight and smoking also avoided. Baseline lower BMI and absence of microalbuminuria were associated with tight control. CONCLUSION: Median difference of HbA1c/BP 1.6%/25/5 mmHg between tight and adverse control considerably reduced the risk of cardiovascular diseases. The findings call for a multi-factorial approach to improve HbA1c, BP, obesity, smoking, and microalbuminuria. PMID- 19679370 TI - Abandoned babies and absent policies. AB - OBJECT: Although infant abandonment is a historical problem, we know remarkably little about the conditions or effects of abandonment to guide evidence driven policies. This paper briefly reviews the existing international evidence base with reference to potential mental health considerations before mapping current UK guidelines and procedures, and available incidence data. Limitations arising from these findings are discussed with reference to international practice, and interpreted in terms of future pathways for UK policy. METHOD: A systematic approach was utilized to gather available data on policy information and statistics on abandoned babies in the UK. RESULTS: A review of the limited literature indicates that baby abandonment continues to occur, with potentially wide-ranging mental health ramifications for those involved. However, research into such consequences is lacking, and evidence with which to understand risk factors or motives for abandonment is scarce. International approaches to the issue remain controversial with outcomes unclear. Our systematic search identified that no specific UK policy relating to baby abandonment exists, either nationally or institutionally. This is compounded by a lack of accurate of UK abandonment statistics. Data that does exist is not comprehensive and sources are incompatible, resulting in an ambiguous picture of UK baby abandonment. CONCLUSIONS: Available literature indicates an absence of clear provision, policy and research on baby abandonment. Based on current understanding of maternal and child mental health issues likely to be involved in abandonment, existing UK strategy could be easily adapted to avoid the 'learning from scratch' approach. National policies on recording and handling of baby abandonments are urgently needed, and future efforts should be concentrated on establishing clear data collection frameworks to inform understanding, guide competent practice and enable successfully targeted interventions. PMID- 19679371 TI - Do baseline glucocorticoids predict fitness? AB - Baseline glucocorticoid (cort) levels are increasingly employed as physiological indices of the relative condition or health of individuals and populations. Often, high cort levels are assumed to indicate an individual or population in poor condition and with low relative fitness (the Cort-Fitness Hypothesis). We review empirical support for this assumption, and find that variation in levels of baseline cort is positively, negatively, or non-significantly related to estimates of fitness. These relationships between levels of baseline cort and fitness can vary within populations and can even shift within individuals at different times in their life history. Overall, baseline cort can predict the relative fitness of individuals and populations, but the relationship is not always consistent or present. PMID- 19679372 TI - Urgent preservation of boreal carbon stocks and biodiversity. AB - Containing approximately one-third of all remaining global forests, the boreal ecosystem is a crucial store of carbon and a haven for diverse biological communities. Historically, fire and insects primarily drove the natural dynamics of this biome. However, human-mediated disturbances have increased in these forests during recent years, resulting in extensive forest loss for some regions, whereas others face heavy forest fragmentation or threat of exploitation. Current management practices are not likely to maintain the attendant boreal forest communities, nor are they adequate to mitigate climate change effects. There is an urgent need to preserve existing boreal forests and restore degraded areas if we are to avoid losing this relatively intact biodiversity haven and major global carbon sink. PMID- 19679374 TI - The beating pattern of the flagellum of Euglena gracilis under altered gravity during parabolic flights. AB - The unicellular freshwater flagellate Euglena gracilis shows negative gravitactic behavior. Previous experiments have revealed that the orientation is most likely an active physiological process in which the beating pattern of the flagellum is controlled by gravity and mediated by a change in the calcium concentration inside the cell. In a signal transduction chain, the calcium signal activates a calmodulin, which in turn raises the concentration of cAMP. This alters the beating pattern of the flagellum; reorientation is therefore not a passive process driven by buoyancy. In a recent parabolic flight experiment (ESA 45th parabolic flight campaign), we observed the beating of the flagellum with a high resolution light microscope. Transition from hyper g to microg as well as from microg to hyper g caused a change of the beating pattern of the flagellum, which confirmed the physiological nature of the process. In microg cells stopped moving the flagellum or tried to reorient, while in hyper g, the cells realigned consecutively. The reaction times for the flagellar responses in previous experiments are confirmed. PMID- 19679373 TI - A mutation within the transmembrane domain of melanosomal protein Silver (Pmel17) changes lumenal fragment interactions. AB - Melanocytes synthesize and store melanin within tissue-specific organelles, the melanosomes. Melanin deposition takes place along fibrils found within these organelles and fibril formation is known to depend on trafficking of the membrane glycoprotein Silver/Pmel17. However, correctly targeted, full-length Silver/Pmel17 cannot form fibers. Proteolytic processing in endosomal compartments and the generation of a lumenal Malpha fragment that is incorporated into amyloid-like structures is also essential. Dominant White (DWhite), a mutant form of Silver/Pmel17 first described in chicken, causes disorganized fibers and severe hypopigmentation due to melanocyte death. Surprisingly, the DWhite mutation is an insertion of three amino acids into the transmembrane domain; the DWhite-Malpha fragment is unaffected. To determine the functional importance of the transmembrane domain in organized fibril assembly, we investigated membrane trafficking and multimerization of Silver/Pmel17/DWhite proteins. We demonstrate that the DWhite mutation changes lipid interactions and disulfide bond-mediated associations of lumenal domains. Thus, partitioning into membrane microdomains and effects on conformation explain how the transmembrane region may contribute to the structural integrity of Silver/Pmel17 oligomers or influence toxic, amyloidogenic properties. PMID- 19679376 TI - A case of epidermolysis bullosa acquisita with clinical features of Brunsting Perry pemphigoid showing an excellent response to colchicine. AB - BACKGROUND: Brunsting-Perry pemphigoid is a rare subepidermal blistering disease characterized by scarring blisters on the head and neck. However, the identity of the responsible autoantigens is still unresolved. METHODS: We reported a patient with epidermolysis bullosa acquisita who had clinical features typical of Brunsting-Perry pemphigoid and investigated the involved type VII collagen epitopes. The patient was a 65-year-old Japanese woman with a 20-month history of recurrent subepidermal bullae on her head, face, and neck. RESULTS: Immunoblot studies revealed that the serum of this patient reacted with type VII collagen, specifically with the noncollagenous domain 1 and the triple-helical domain. The patient responded completely to colchicine monotherapy. LIMITATIONS: This study was performed on only one case. CONCLUSION: This study suggests that Brunsting Perry pemphigoid may be a clinical variant of epidermolysis bullosa acquisita. PMID- 19679375 TI - Accuracy of teledermatology for pigmented neoplasms. AB - BACKGROUND: Accurate diagnosis and management of pigmented lesions is critical because of the morbidity and mortality associated with melanoma. OBJECTIVE: We sought to compare accuracy of store-and-forward teledermatology for pigmented neoplasms with standard, in-person clinic dermatology. METHODS: We conducted a repeated measures equivalence trial involving veterans with pigmented skin neoplasms. Each lesion was evaluated by a clinic dermatologist and a teledermatologist; both generated a primary diagnosis, up to two differential diagnoses, and a management plan. The primary outcome was aggregated diagnostic accuracy (match of any chosen diagnosis with histopathology). We also compared the severity of inappropriately managed lesions and, for teledermatology, evaluated the incremental change in accuracy when polarized light dermatoscopy or contact immersion dermatoscopy images were viewed. RESULTS: We enrolled 542 patients with pigmented lesions, most were male (96%) and Caucasian (97%). The aggregated diagnostic accuracy rates for teledermatology (macro images, polarized light dermatoscopy, and contact immersion dermatoscopy) were not equivalent (95% confidence interval for difference within +/-10%) and were inferior (95% confidence interval lower bound <10%) to clinic dermatology. In general, the addition of dermatoscopic images did not significantly change teledermatology diagnostic accuracy rates. In contrast to diagnostic accuracy, rates of appropriate management plans for teledermatology were superior and/or equivalent to clinic dermatology (all image types: all lesions, and benign lesions). However, for the subgroup of malignant lesions (n = 124), the rate of appropriate management was significantly worse for teledermatology than for clinic dermatology (all image types). Up to 7 of 36 index melanomas would have been mismanaged via teledermatology. LIMITATIONS: Nondiverse study population and relatively small number of melanomas were limitations. CONCLUSIONS: In general, the diagnostic accuracy of teledermatology was inferior whereas management was equivalent to clinic dermatology. However, for the important subgroup of malignant pigmented lesions, both diagnostic and management accuracy of teledermatology was generally inferior to clinic dermatology and up to 7 of 36 index melanomas would have been mismanaged via teledermatology. Teledermatology and teledermatoscopy should be used with caution for patients with suspected malignant pigmented lesions. PMID- 19679377 TI - Oxidative stress activates the pro-survival TrkA pathway through membrane cholesterol loss. AB - Neuronal activity is a highly demanding energetic process, resulting in the gradual accumulation of reactive oxygen species (ROS). Despite comparatively weak anti-oxidant defence systems, neurons outlive the pressure of ROS by activating most robust anti-stress mechanisms. We recently showed that one such mechanism is the activation of the TrkB receptor pathway, in turn determined by the loss of membrane cholesterol. It is not known however what causes the loss of membrane cholesterol. We here show that in differentiated PC12 cells induction of ROS is paralleled by a moderate loss of membrane cholesterol and the activation of the pro-survival TrkA receptor. Pharmacological reduction of cholesterol in non stressed cells triggers TrkA activation while cholesterol replenishment inhibits receptor activation induced by stress. Moreover, addition of a ROS inhibitor prevented cholesterol loss and receptor activation under stress. These results highlight cholesterol loss as a compensatory protective mechanism against acute stress. PMID- 19679378 TI - Pentacyclo-undecane derived cyclic tetra-amines: synthesis and evaluation as potent anti-tuberculosis agents. AB - As part of an ongoing effort to develop highly potent anti-tuberculosis agents, fourteen pentacyclo-undecane (PCU) tetra-amine compounds were synthesized and screened for their in vitro anti-mycobacterial activity against two TB strains, H37Rv and XDR 194 [an extensively drug-resistant strain of tuberculosis]. Using the broth macrodilution method, nitrofuranylamide based compounds (6a and 6b) showed almost similar activities against the H37Rv strain of Mycobacterium tuberculosis when compared with the control drug, ethambutol. N-Geranyl piperazine PCU (8a) and trans-trans farnesyl piperazine PCU (8b) were 3.2 and 3.7 times more potent than commercially available ethambutol. Both isoprenyl PCU tetra-amine derivatives and N-decyl piperazine PCU (9a) were highly active against the XDR 194 strain of tuberculosis with MICs in the range of 0.63-3.02 microM. Cytotoxicities (IC(50)) of isoprenyl based compounds (8a, 8b) and compound 9a were tested on a mammalian cell line [MDBK (Madin Darby bovine kidney epithelium)] with values of 30, 24 and 25 microM respectively. PMID- 19679379 TI - Synthesis and preliminary pharmacological evaluation of imidazo[2,1-f]purine-2,4 dione derivatives. AB - A series of N-8-arylpiperazinylpropyl derivatives of 1,3-dimethyl-(1H,8H) imidazo[2,1-f]purine-2,4-dione (2-10) and amide derivatives of 1,3-dimethyl-6,7 dihydroimidazo[2,1-f]purine-2,4-(1H,3H)-dione-7-carboxylic acid (11-13) were synthesized. Compounds (2-10) evaluated in vitro were potent 5-HT(1A) receptor ligands. Preclinical studies indicated that 8-[3-(N4-phenyl)-piperazin-N1-yl propyl]-1,3-dimethyl-(1H,8H)-imidazo[2,1-f]purine-2,4-dione (2) exerts anxiolytic like activity in the four-plate test in mice; however its effect was weaker, than that produced by Diazepam. This compound and 8-[3-(N4-2'-metoxyphenyl)-piperazin N1-yl-propyl]-1,3-dimethyl-(1H,8H)-imidazo[2,1-f]purine-2,4-dione (3) behaved like antidepressants in the forced swimming test in mice; and their activity in that model was comparable with the effect of Imipramine. The obtained results suggested that the long-chain arylpiperazines (LCAPs) linked to tricyclic derivatives of the theophylline remain a worthy of future research for obtaining new derivatives with potential anxiolytic/antidepressant activity. PMID- 19679381 TI - Variability of motion in individuals with mechanical or functional ankle instability during a stop jump maneuver. AB - BACKGROUND: Movement variability may influence episodes of instability following lateral ankle sprain. METHODS: Sixty-three recreational athletes with a history of moderate-severe ankle sprain were recruited. Mechanically and functionally unstable ankle groups had 2 episodes of instability in the last year. Mechanically unstable had clinically lax lateral ankle ligaments; functionally unstable and copers did not. Copers had a history of sprain but no residual instability. Lower extremity 3-dimensional kinematics and ground reaction forces were measured during a 2-legged stop jump. Average ensemble curves of eight trials normalized to 100% of stance phase were created. The coefficient of variation and average standard deviation of the ensemble curves of each variable were identified. A log(e) (ln) transformation was performed on the data. One-way ANOVAs with Tukey post hoc testing were utilized with alpha=0.05. FINDINGS: The functionally unstable group demonstrated greater mean (standard deviation) ln coefficient of variation ankle inversion/eversion 3.56 (1.19) than the mechanically unstable 2.77 (0.95) and copers 2.74 (1.05) (P=0.05 and P=0.04; eta(p)(2)=0.12), and greater ln standard deviation ankle inversion/eversion 1.07 (0.78) than copers 0.61 (0.31) (eta(p)(2)=0.13). The mechanically unstable group demonstrated greater ln coefficient of variation anterior-posterior ground reaction force 3.69 (0.27) than functionally unstable 3.43 (0.25) (P=0.02; eta(p)(2)=0.13). INTERPRETATION: Functionally unstable individuals demonstrated greater ankle frontal plane movement variability during a stop jump, which may increase risk of instability. Mechanically unstable participants demonstrated greater anterior-posterior ground reaction force variability, which may indicate difficulty mitigating landing forces with lax ligaments. Movement variability may influence episodes of ankle instability. PMID- 19679380 TI - Design and one-pot synthesis of alpha-aminophosphonates and bis(alpha aminophosphonates) by iron(III) chloride and cytotoxic activity. AB - In this study, we used a solution of FeCl(3) in THF to facilitate the Mannich type reaction of aldehyde, amine and phosphite compounds to form corresponding alpha-aminophosphonates in a one-pot, three-component reaction. Selected alpha aminophosphonates were entered into a biological assay test and were studied by docking methods, using Autodock 3.0. The results showed that the reactions were carried out mildly and eco-friendly to form alpha-aminophosphonates in high yields. Some were found to have cytotoxic activity on the cell lines RAJI, JURKAT and MCF-7. An indole derived bis(alpha-aminophosphonates) showed maximum cytotoxic effect comparable to doxorubicin. Although the FDE (Final Docking Energy) for the most cytotoxic compound was of the most negative value, there is no correlation between FDE and cytotoxicity. PMID- 19679382 TI - Validation of regression models for nitrate concentrations in the upper groundwater in sandy soils. AB - For Dutch sandy regions, linear regression models have been developed that predict nitrate concentrations in the upper groundwater on the basis of residual nitrate contents in the soil in autumn. The objective of our study was to validate these regression models for one particular sandy region dominated by dairy farming. No data from this area were used for calibrating the regression models. The model was validated by additional probability sampling. This sample was used to estimate errors in 1) the predicted areal fractions where the EU standard of 50 mg l(-1) is exceeded for farms with low N surpluses (ALT) and farms with higher N surpluses (REF); 2) predicted cumulative frequency distributions of nitrate concentration for both groups of farms. Both the errors in the predicted areal fractions as well as the errors in the predicted cumulative frequency distributions indicate that the regression models are invalid for the sandy soils of this study area. PMID- 19679383 TI - Combined effects of elevated CO2 and natural climatic variation on leaf spot diseases of redbud and sweetgum trees. AB - Atmospheric CO(2) concentrations are predicted to double within the next century and alter climate regimes, yet the extent that these changes will affect plant diseases remains unclear. In this study conducted over five years, we assessed how elevated CO(2) and interannual climatic variability affect Cercospora leaf spot diseases of two deciduous trees. Climatic data varied considerably between the five years and altered disease expression. Disease incidence and severity for both species were greater in years with above average rainfall. In years with above average temperatures, disease incidence for Liquidambar styraciflua was decreased significantly. When significant changes did occur, disease incidence and severity always increased under elevated CO(2). Chlorophyll fluorescence imaging of leaves revealed that any visible increase in disease severity induced by elevated CO(2) was mitigated by higher photosynthetic efficiency in the remaining undamaged leaf tissue and in a halo surrounding lesions. PMID- 19679384 TI - Neural correlates of traditional Chinese medicine induced advantageous risk taking decision making. AB - This fMRI study examined the neural correlates of the observed improvement in advantageous risk-taking behavior, as measured by the number of adjusted pumps in the Balloon Analogue Risk Task (BART), following a 60-day course of a Traditional Chinese Medicine (TCM) recipe, specifically designed to regulate impulsiveness in order to modulate risk-taking behavior. The 14 participants recruited for this study were randomly assigned to the experimental and control groups and the TCM recipe (Panax, 520 mg; Astragalus membranaceous Bunge, 520 mg; Masnetitum, 840 mg; Ostrea gigas Thumb, 470 mg; Thinleaf Milkwort Root Radix Polygalae, 450 mg; and Os Draconis, 470 mg) was administered, as a diet supplement, to the seven participants in the experimental group. The neural activity of the two groups was monitored by a 3T MRI scanner, before and after the 60-day treatment. Associated with the improved advantageous risk-taking behavior seen in the experimental group, significantly stronger blood oxygenation level dependent (BOLD) responses were observed in the bilateral dorsolateral prefrontal cortex (DLPFC), left putamen, left thalamus, right insula, and right anterior cingulate cortex (ACC), regions which have previously been reported as being involved in risk-taking decision making. The effect of the TCM in improving advantageous risk-taking decision making appears to have been related to the enhanced efficiency of the cognitive affective system, the PFC-ACC-insula-striatum network, which functions to inhibit impulsiveness, to sensitize reward-related information, and to allow the opportunity, during risk estimation, to evaluate potential gains and losses. The findings of this study suggest that interventions acting on factors modulating risk-taking decision making could have a beneficial effect in terms of optimizing risk-taking behavior. PMID- 19679386 TI - [Treatment by fat tissue transfer for radiation injury in childhood facial cancer]. AB - The radiation treatment of malignant facial tumors in children may induce major functional and cosmetic sequelae, mainly due to uneven growth of the bones and soft tissues, resulting in facial asymmetry and hemihypotrophy at adult age. Although fat transfer has proven effective for facial cosmetic treatment, few studies have demonstrated the benefit of the technique in heavily irradiated tissues. The techniques generally used for the treatment of facial asymmetry or hypotrophy are ill-adapted to irradiated patients. Indeed, procedures such as skin detachment, osteotomy and vascular suture are risky because of radiation induced damage. The aim of the present study was to investigate the potential benefits of fat transfer for the correction of sequelae of facial irradiation. Four patients (two males and two females) aged 27, 25, 16 and 13 years underwent fat grafting for the correction of facial asymmetry or hypotrophy induced by cancer radiation treatment during childhood (radiation dose of more than 50Gy). One to three grafting sessions were required, depending on the cases. After a median follow-up of 3.9 years, cosmetic results were considered satisfactory by both the patient and the surgeon in all four cases. Fat transfer remarkably improved the cosmetic appearance of the patients, without deleterious consequences for the vitality of tissues. In addition, a restoration of skin trophicity was observed, thus confirming the benefit of grafting adipocytes into the irradiated integument. In conclusion, fat grafting appears to be a simple and easily reusable technique which makes it possible to obtain the best morphological and cosmetic results in irradiated patients, whereas avoiding complex and potentially hazardous procedures. PMID- 19679385 TI - [Prevention of seroma by quilting suture after harvesting latissimus dorsi flap. The "Chippendale" technic]. AB - Seroma is the most frequent minor complication after harvesting latissimus dorsi flap for breast reconstruction. It induces patient's discomfort and multiple consultations for punctions. The dead space resulting from the harvest has to be closed by the "quilting suture" in order to prevent the seroma. Our aim is to evaluate the efficiency and the tolerance of the quilting suture by comparing two groups of 100 patients who had a breast reconstruction by the same technic of extended latissimus dorsi flap, performed by the same surgeon, from 2004 to 2007. Half of patients had the classic way of dorsal closure, and the other half of patients had the dorsal quilting suture. PATIENTS AND METHODS: In order to compare the two groups we have collected data concerning age, body mass index (BMI), tobacco use, postoperative complications, number and volume of punctions, draining time and postoperative pain. The efficiency of the quilting suture lies on a rigorous repartition of at least six sutures on the upper skin flap, 12 on the lower skin flap and under the skin suture line. The suture model is based on the one used for the Chippendale-designed sofa. We suture the skin flap while pushing down the shoulder, in order to split the skin tension and avoid traction on the final skin suture line. The procedure takes 15 minutes. RESULTS: The "Chippendale" technic allows to reduce draining time from 12 days to 6 days. The incidence of chronic seroma is reduced by 50%. The dorsal wound healing seems also better thanks to tension reduction resulting from the quilting suture. CONCLUSION: The "Chippendale" technic is a quick, cheap and easy learned procedure, efficient for preventing chronic seroma after the latissimus dorsi flap. The postoperative recovery is eased and the patients comforted. PMID- 19679387 TI - Excess risk of stillbirth during the 1918-1920 influenza pandemic in Japan. PMID- 19679388 TI - Extracorporeal shock waves stimulate osteoblast activities. AB - The extracorporeal shock wave therapy (ESWT) is an extensively applied treatment for musculoskeletal disorders because it promotes bone repair. The aim of this study was to evaluate the direct effect of ESWT on murine osteoblasts to clarify the cellular mechanism that leads to the induction of osteogenesis. Osteoblasts in culture flasks were treated with ESWT pulses (500 impulses of 0.05 mJ/mm(2)) generated by an electromagnetic device. Using western blot analysis 3h after ESWT, an increased expression of Bax was found, indicating a fast pro-apoptotic effect of treatment on some of the osteoblasts. Activation of the cyclin E2/CDK2 is the complex that regulates the G1-S transition and is essential for cell proliferation. It was evident 24 to 72h after treatment, indicating a proliferative stimulus. A decreased expression of osteoprotegerin (OPG) and receptor activator NF kappa B ligand (RANKL) 24 and 48h after ESW, followed by a later increase of OPG, paired with a much smaller increase of RANKL, was evident by real-time polymerase chain reaction (PCR). The decreased RANKL/OPG ratio suggests inhibition of osteoclastogenesis. We can conclude that ESWT induces bone repair through the proliferation and differentiation of osteoblasts and the reduction of their secretion of pro-osteoclastogenic factors. PMID- 19679389 TI - The effect of back reflections on the acoustic power delivered by physiotherapy ultrasound machines. AB - Physiotherapy ultrasound is used widely for the treatment of soft tissue injuries. The ultrasonic treatment heads are all highly resonant devices and may therefore be sensitive to the levels of acoustic back reflection that they experience. However, the extent to which reflections affect acoustic power during clinical use has not been reported in the literature and is not addressed in current technical standards. This study investigated the effect of physiological levels of acoustic reflection on 29 physiotherapy treatment heads from a total of 21 machines and 6 manufacturers. A range of membranes were constructed and used to mimic the levels of acoustic reflections that occur during treatment. The results obtained showed that almost half of the heads tested (45%) had deviations in acoustic power of more than 15% compared with free-field measurements. Four heads (17%) had deviations in power of more than 25%. We recommend that the susceptibility of physiotherapy ultrasound machines to acoustic reflections be addressed in the relevant technical standards. Also, it is appropriate for tests to be carried out during design and manufacture, and by the purchaser during their acceptance testing. PMID- 19679390 TI - High-frequency ex vivo ultrasound imaging of the auditory system. AB - A 50MHz array-based imaging system was used to obtain high-resolution images of the ear and auditory system. This previously described custom built imaging system (Brown et al. 2004a, 2004b; Brown and Lockwood 2005) is capable of 50 microm axial resolution, and lateral resolution varying from 80 microm to 130 microm over a 5.12 mm scan depth. The imaging system is based on a 2mm diameter, seven-element equal-area annular array, and a digital beamformer that uses high speed field programmable gate arrays (FPGAs). The images produced by this system have shown far superior depth of field compared with commercially available single-element systems. Ex vivo, three-dimensional (3-D) images were obtained of human cadaveric tissues including the ossicles (stapes, incus, malleus) and the tympanic membrane. In addition, two-dimensional (2-D) images were obtained of an intact cochlea by imaging through the round window membrane. The basilar membrane inside the cochlea could clearly be visualized. These images demonstrate that high-frequency ultrasound imaging of the middle and inner ear can provide valuable diagnostic information using minimally invasive techniques that could potentially be implemented in vivo. PMID- 19679391 TI - Lanthanum citrate induces anoikis of Hela cells. AB - Some reports show that lanthanum, a rare earth element, induces apoptosis in certain cancer cells. In the present paper, we report the first observation of anoikis induced by lanthanum citrate (LaCit) in Hela cells at a concentration of 0.001-0.1 mmol/L after 48h-treatment. Before cell treatment, Hela cells were subjected to anoikis-resistant selection to remove anoikis-sensitive cells and ensure specificity of LaCit-induced anoikis. Anoikis was determined by Annexin/PI, AO/EB staining, cleavage of PARP and soft-agar colony forming assay. Further, findings of decreased mitochondrial membrane potential, the cleavage of caspase-9 and a dose-dependent increase expression of Bax were detected, suggesting that the intrinsic caspase pathway was involved in the anoikis induced by LaCit. In addition, activation of caspase-8 occurred later than that of caspase-9. LaCit also caused reorganization of actin cytoskeleton, and was accompanied by an increase in co-localization of F-actin with mitochondria, implying that both actin cytoskeleton and mitochondria may play important roles in LaCit -induced anoikis. PMID- 19679392 TI - Partitioning characteristics and particle size distributions of heavy metals in the O2/RFG waste incineration system. AB - This study applies the oxygen/recycled flue gas (O(2)/RFG) combustion technology for waste incineration and investigates the effects of different RFG rates on the concentrations of gas pollutants as well as the partitioning characteristics and particle size distributions of five heavy metals (Cr, Cu, Pb, Zn, and Cd). Experimental results show that the combustion efficiency can be improved and the concentration of CO(2) is increased by appropriately controlling the RFG rate in the O(2)/RFG incineration system. The partitioning characteristics and size distributions of heavy metals in O(2)/RFG combustion system are different from those in general air combustion system. Under O(2)/RFG combustion system, the partitioning percentages of heavy metals in sand bed, bottom ash, and collected ash are increased. The particle size distributions of heavy metals display the triple-peak curves. As the RFG rate rises, the concentrations of volatile heavy metals are increased in small-size (<1 microm) fly ash, decreased in medium-size (1-10 microm) fly ash and increased in large-size (>10 microm) fly ash. These phenomena will benefit to increase the overall control efficiency of heavy metals in the incineration system. PMID- 19679393 TI - More than a decade of conflict between hazardous waste management and public resistance: a case study of NIMBY syndrome in Souselas (Portugal). AB - Portugal's export amount of hazardous waste is increasing. More than 10 years ago, waste co-incineration in cement kilns was proposed in Portugal for technical and economic reasons amid administrative willingness to manage hazardous waste domestically. However, this waste project has still not been realized owing to local public resistance (the so-called NIMBY syndrome). We focus attention on the long-term resistance, and the following points are established through analysis: (i) public participation was left out of the project at the initial stage, so public confidence in government ability has been declining, (ii) public antipathy results from emotive stimulation and/or mental fears rather than scientific evidence, and (iii) indirect socio-economic factors in the region proposed for a hazardous waste facility are completely excluded from the project scope. The presented case study suggests that public acceptability is quite important in implementing hazardous waste management without delay. Therefore, engineers, researchers and planners in hazardous waste management should be aware that a report addressing only technical performance is less beneficial than a comprehensive report for putting a management tool into industrial practice. PMID- 19679394 TI - Simple and rapid spectrophotometric determination of trace titanium (IV) enriched by nanometer size zirconium dioxide in natural water. AB - A novel method for preconcentration of Ti(IV) with nanometer size ZrO(2) and determination by spectrophotometry has been developed. Ti(IV) was selectively adsorbed on 300 mg ZrO(2) from 500 mL solution at pH 6.0, then eluted by 5 mL 11.3 mol L(-1) HF. The eluent added was diantipyrylmethane (DAPM, as chromogenic reagent) and ascorbic acid (as masking agent), used for the analysis of Ti(IV) by measuring the absorbance at 390 nm with spectrophotometry, based on the chromogenic reaction between the Ti(IV) and DAPM. This method gave a concentration enhancement of 100 for 500 mL sample, eliminated the sizable interferences on direct determination with spectrophotometry. Detection limit (3 sigma, n=11) of 0.1 microg L(-1) was obtained. The method was applied to determine the concentration of Ti(IV) in river water and seawater and the analytical recoveries of Ti(IV) added to samples were 97.6-101.3%. PMID- 19679395 TI - Are we getting anywhere in human pain genetics? PMID- 19679396 TI - Hypothalamic deep-brain stimulation modulates thermal sensitivity and pain thresholds in cluster headache. AB - Deep-brain stimulation (DBS) of the posterior hypothalamus has been shown to be clinically effective for drug-resistant chronic cluster headache, but the underlying mechanism is still not understood. The hypothalamus as an important centre of homeostasis is connected among others to the trigeminal system via the trigeminohypothalamic tract. We aimed to elucidate whether hypothalamic stimulation affects thermal sensation and pain perception only in the clinically affected region (the first trigeminal branch) or in other regions as well. Thus, we examined three groups: chronic cluster headache patients with unilateral DBS of the posterior hypothalamus (n = 11), chronic cluster headache patients without DBS (n = 15) and healthy controls (n = 29). Perception and pain thresholds for hot and cold stimuli were determined bilaterally in all subjects supraorbitally, at the forearm, and in the lower leg. In DBS patients, thresholds were determined with the stimulator activated and inactivated. Cold pain thresholds at the first trigeminal branch were increased on the stimulated side in the DBS group compared to healthy subjects (p = .015). The DBS group also had higher cold detection thresholds compared to non-implanted cluster headache patients (p < .05). Short term interruption of stimulation did not induce any changes in DBS patients. Clinically relevant differences were found neither between non-stimulated cluster headache patients and healthy controls nor between the affected and the non affected sides in the chronic cluster headache patients without DBS. These results support the notion that neurostimulation of the posterior hypothalamus is specific for cluster headache and only affects certain aspects of pain sensation. PMID- 19679397 TI - Clinicopathological findings, molecular detection and characterization of Babesia gibsoni infection in a sick dog from Italy. AB - A 4-year-old intact female American Pit Bull Terrier from Italy descendant of an American-born bitch was evaluated for anorexia, lethargy, weakness, and intermittent vomiting. On physical examination, the dog was dehydrated, had pale mucous membranes, hunched posture and abdominal pain. A moderate anemia was observed. Splenomegaly and hyperechoic regions suspected as infarcts in the spleen were seen on abdominal ultrasound. Based on the suspicion of splenic torsion, splenectomy was performed. After surgery, the clinical condition deteriorated. A follow-up complete blood count demonstrated severe macrocytic normochromic anemia with evidence of marked regeneration, left shift neutrophilia, monocytosis and marked thrombocytopenia. Blood smear evaluation revealed single to multiple, variable sized (1-3 microm in diameter), and round to oval to band-like piroplasms within many red blood cells consistent with small form Babesia spp. or Theileria spp. A partial segment of the 18S rRNA gene was amplified and the PCR product was analyzed by direct sequencing. The nucleotide sequence was completely identical to that of Babesia gibsoni present in GenBank. This is the first molecular detection and characterization of B. gibsoni infection in a sick dog from Italy. PMID- 19679398 TI - Sources of potentially infectious human microsporidia: molecular characterisation of microsporidia isolates from exotic birds in the Czech Republic, prevalence study and importance of birds in epidemiology of the human microsporidial infections. AB - A total of 287 faecal specimens of captive exotic birds from the orders Psittaciformes, Passeriformes and Columbiformes were randomly collected from Bohemian pet stores, avian breeders and avian keepers and were screened for the presence of human pathogenic microsporidia by polymerase chain reaction (PCR). Microsporidial DNA was identified in 115 faecal samples (40.1%). Single-species infection was detected in 36 birds (12.5%) for Enterocytozoon bieneusi, 36 birds (12.5%) for Encephalitozoon cuniculi and 18 birds (6.3%) for Encephalitozoon hellem. No Encephalitozoon intestinalis positive samples were identified. Moreover, co-infections were detected in 25 birds: E. bieneusi together with E. cuniculi in 14 animals (4.9%) or E. hellem in 11 cases (3.8%). E. hellem was present in 1A (5.2%) and three (0.3%) genotypes, E. cuniculi in I (2.4%), II (8.0%) and III (0.7%) genotypes and E. bieneusi in A (8.4%) and EbpA (10.8%) genotypes. Several of these genotypes have never been recorded in birds before. The results of this report suggest the low host specificity of E. bieneusi, E. hellem and E. cuniculi and describe 44 new avian hosts. PMID- 19679399 TI - Is recurrent epistaxis from Kiesselbach's area (REKAS) in any relationship to the hemorrhoidal disease? AB - Vascular diseases are a major threat to human health nowadays. Hypertension, cardiovascular disease and varicose vain disease including hemorrhoids, are now increasingly recognized as inflammatory diseases. The role of inflammation cytokines in the pathogenesis of these diseases is very important. The lamina propria in the nasal mucosa is rich in blood vessels and humoral mediators. Recurrent epistaxis from Kiesselbach's area syndrome (REKAS) was first mentioned as early as 1985. It has been found that 90% of patients suffering from recurrent epistaxis from Kiesselbach area syndrome simultaneously suffered from hemorrhoids. Clinical observations suggest a possible mutual pathophysiologic relationship between Kiesselbach's and anorectal venous plexus. This relationship is also suggested in the reverse direction: significantly more than two thirds of primarily hemorrhoidal patients (83.01%) showed simultaneous vascular dilatations within their Kiesselbach plexuses, but none of these patients had ever have recurrent nose bleeds. There is one more thing they did not have (contrary to REKAS group)--anterior septal deformity. Furthermore, REKAS and hemorrhoidal disease, despite being different clinical entities, frequently appear in the primarily REKAS patients or their closest relatives (more than 90% out of all!). At the same time, all of REKAS patients did have a certain degree of the anterior septal deformity, which primarily hemorrhoidal patients did not have at all. Therefore we presume that Kiesselbach's vascular plexus in the Little's area of the nasal septum belongs to the same group as anorectal venous plexus does (others of this group are brain, esophagus, and lower leg venous system). We also presume that the anterior septal deformity is a crucial factor for the onset of the inflammation of the nasal vestibule skin (vestibulitis nasi), while vestibulitis nasi precipitates the onset of typical recurrent nose bleeds from the Kiesselbach's plexus. PMID- 19679401 TI - Pulmonary hypertensive necrotizing arteritis: an unusual case of unilateral involvement in an infant with congenital heart disease. AB - Pulmonary hypertension is clinically defined as a pulmonary arterial pressure greater than 25 mmHg at rest or greater than 30 mmHg during exercise. We report a unique case of an infant with congenital heart disease causing pulmonary hypertension and necrotizing pulmonary arteritis but affecting only one lung. In conclusion, in pulmonary valve atresia, the lungs may be supplied via aortopulmonary collaterals. Necrotizing pulmonary arteritis is a severe lesion seen in irreversible pulmonary hypertension. PMID- 19679400 TI - Frequency and type of KRAS mutations in routine diagnostic analysis of metastatic colorectal cancer. AB - Mutation analysis of the KRAS oncogene is now established as a predictive biomarker in colorectal cancer (CRC). Large prospective clinical trials have shown that only CRCs with wild-type KRAS respond to anti-epidermal growth factor receptor (EGFR) treatment. Therefore, mutation analysis is mandatory before treatment, and reliable benchmarks for the frequency and types of KRAS mutations have to be established for routinely testing large numbers of metastatic CRCs. A thousand and eighteen cases (879 primary tumors and 139 metastases) of metastatic colorectal cancer were analyzed for the KRAS mutational status of codons 12 and 13 of the KRAS gene by genomic sequencing in a routine setting. Results were analyzed separately for specimens derived from primary tumors and metastases. KRAS mutations in codons 12 and 13 were present in 39.3% of all analyzed CRCs. The most frequent types of mutations were glycine to aspartate on codon 12 (p.G12D, 36.0%), glycine to valine on codon 12 (pG12V, 21.8%), and glycine to aspartate on codon 13 (p.G13D, 18.8%). They account for 76.6% of all mutations and prevail in primary tumors and distant metastases, indicating a robustness of the KRAS mutational status during neoplastic dissemination. The frequency of KRAS mutations and the preponderance of three types of mutations in codons 12 and 13 in a large, unselected cohort of metastatic CRC confirm the previous data of small and selected CRC samples. Thus, a mutation frequency of 40% and a cluster of three mutation types (p.G12D, pG12V, and p.G13D) in primaries and metastases can be defined as benchmarks for routine KRAS analyses. PMID- 19679402 TI - In vitro cytotoxicity of low-dose-rate radioimmunotherapy by the alpha-emitting radioimmunoconjugate Thorium-227-DOTA-rituximab. AB - PURPOSE: To determine whether the low-dose-rate alpha-particle-emitting radioimmunoconjugate (227)Th-1,4,7,10-p-isothiocyanato-benzyl tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA)-rituximab can be used to inactivate lymphoma cells growing as single cells and small colonies. METHODS AND MATERIALS: CD20-positive lymphoma cell lines were treated with (227)Th-DOTA rituximab for 1-5 weeks. To simulate the in vivo situation with continuous but decreasing supply of radioimmunoconjugates from the blood pool, the cells were not washed after incubation with (227)Th-DOTA-rituximab, but half of the medium was replaced with fresh medium, and cell concentration and cell-bound activity were determined every other day after start of incubation. A microdosimetric model was established to estimate the average number of hits in the nucleus for different localizations of activity. RESULTS: There was a specific targeted effect on cell growth of the (227)Th-DOTA-rituximab treatment. Although the cells were not washed after incubation with (227)Th-DOTA-rituximab, the average contribution of activity in the medium to the mean dose was only 6%, whereas the average contribution from activity on the cells' own surface was 78%. The mean dose rates after incubation with 800 Bq/mL (227)Th-DOTA-rituximab varied from 0.01 to 0.03 cGy/min. The average delay in growing from 10(5) to 10(7) cells/mL was 15 days when the cells were treated with a mean absorbed radiation dose of 2 Gy alpha-particle radiation from (227)Th-DOTA-rituximab, whereas it was 11 days when the cells were irradiated with 6 Gy of X-radiation. The relative biologic effect of the treatment was estimated to be 2.9-3.4. CONCLUSIONS: The low-dose rate radioimmunoconjugate (227)Th-DOTA-rituximab is suitable for inactivation of single lymphoma cells and small colonies of lymphoma cells. PMID- 19679403 TI - Improved survival with radiation therapy in high-grade soft tissue sarcomas of the extremities: a SEER analysis. AB - PURPOSE: The benefit of radiation therapy in extremity soft tissue sarcomas remains controversial. The purpose of this study was to determine the effect of radiation therapy on overall survival among patients with primary soft tissue sarcomas of the extremity who underwent limb-sparing surgery. METHODS AND MATERIALS: A retrospective study from the Surveillance, Epidemiology, and End Results (SEER) database that included data from January 1, 1988, to December 31, 2005. A total of 6,960 patients constituted the study population. Overall survival curves were constructed using the Kaplan-Meir method and for patients with low- and high-grade tumors. Hazard ratios were calculated based on multivariable Cox proportional hazards models. RESULTS: Of the cohort, 47% received radiation therapy. There was no significant difference in overall survival among patients with low-grade tumors by radiation therapy. In high-grade tumors, the 3-year overall survival was 73% in patients who received radiation therapy vs. 63% for those who did not receive radiation therapy (p < 0.001). On multivariate analysis, patients with high-grade tumors who received radiation therapy had an improved overall survival (hazard ratio 0.67, 95% confidence interval 0.57-0.79). In patients receiving radiation therapy, 13.5% received it in a neoadjuvant setting. The incidence of patients receiving neoadjuvant radiation did not change significantly between 1988 and 2005. CONCLUSIONS: To our knowledge, this is the largest population-based study reported in patients undergoing limb-sparing surgery for soft tissue sarcomas of the extremities. It reports that radiation was associated with improved survival in patients with high-grade tumors. PMID- 19679404 TI - Xerostomia after treatment for oral and oropharyngeal cancer using the University of Washington saliva domain and a Xerostomia-Related Quality-of-Life Scale. AB - PURPOSE: The first aim of this study was to identify which clinical factors are associated with xerostomia in patients after treatment for oral and oropharyngeal squamous cell carcinoma, using the Xerostomia-Related Quality-of-Life Scale (XeQoLS) and the University of Washington Quality-of-Life Questionnaire Version 4 dry mouth item (UW-QOL v4). The second aim was to compare these two questionnaires and postulate a cutoff in the UW-QOL below which patients are doing sufficient badly to warrant further evaluation and support. METHODS AND MATERIALS: In all, 371 patients alive and disease free treated between 1992 and 2005 were sent the survey, of whom 250 (67%) responded. Various clinical factors correlated with xerostomia, particularly adjuvant radiotherapy and Pstage. RESULTS: In logistic regression analyses to predict three or more problems on the XeQoLS, only adjuvant radiotherapy (p < 0.001) was significant at the 5% level. There were significant (p < 0.001) correlations between the XeQoLS scores (total average and domain) with all the UW-QOL domain scores, the strongest with swallowing (-0.69), taste (-0.64), chewing (-0.64), mood (-0.60), and saliva ( 0.59) domains. Patients scoring <70 (i.e., 0 or 30) on the UW-QOL could be used as a screening cutoff because it formed 1 in 5 of all patients (49/242) but accounted for half (299/566) of the significant problems generated by the XeQoLS. This also identified 13/21 patients with 10 or more problems. CONCLUSION: The UW QOL saliva domain seems to be a suitable means of screening for dry mouth in head and-neck clinics and could be used to trigger interventions. PMID- 19679405 TI - A prospective study of intrafraction prostate motion in the prone vs. supine position. AB - PURPOSE: To prospectively analyze prostate intrafraction motion in the prone vs. supine position and to assess patient satisfaction with these two positions. METHODS AND MATERIALS: Fifteen prostate cancer patients underwent implantation of five fiducial gold seeds in their prostate for localization. Patients were treated with high-dose-rate brachytherapy to 2,200 cGy followed by intensity modulated radiation therapy (IMRT) to 5,040 cGy. Patients underwent computed tomography simulation and IMRT in the prone position. For the first five IMRT treatments, an electronic portal imaging system was used to acquire anteroposterior (AP) and lateral images pretreatment and posttreatment. We then repositioned each patient supine and repeated the process, resulting in 600 images. RESULTS: Mean +/- standard deviation intrafraction prostate motion was 2.1 +/- 1.2 mm and 1.7 +/- 1.4 mm (AP, p = 0.47), 2.2 +/- 2.0 mm and 1.6 +/- 1.8 mm (superoinferior, p = 0.16), and 1.0 +/- 1.2 mm and 0.6 +/- 0.9 mm (left-right, p = 0.03) in the prone and supine positions, respectively. Eighty percent of patients stated that they were more comfortable in the supine position (p = 0.02). CONCLUSIONS: Prone and supine positions resulted in a similar magnitude of AP and superoinferior intrafraction prostate motion (2 mm). Because there was no significant difference in the magnitude of AP and superoinferior prostate motion prone vs. supine and patients were more comfortable in the supine position, patients now undergo IMRT to the prostate and seminal vesicles at our center in the supine position. PMID- 19679406 TI - Simple carotid-sparing intensity-modulated radiotherapy technique and preliminary experience for T1-2 glottic cancer. AB - PURPOSE: To investigate the dosimetry and feasibility of carotid-sparing intensity-modulated radiotherapy (IMRT) for early glottic cancer and to report preliminary clinical experience. METHODS AND MATERIALS: Digital Imaging and Communications in Medicine radiotherapy (DICOM-RT) datasets from 6 T1-2 conventionally treated glottic cancer patients were used to create both conventional IMRT plans. We developed a simplified IMRT planning algorithm with three fields and limited segments. Conventional and IMRT plans were compared using generalized equivalent uniform dose and dose-volume parameters for in-field carotid arteries, target volumes, and organs at risk. We have treated 11 patients with this simplified IMRT technique. RESULTS: Intensity-modulated radiotherapy consistently reduced radiation dose to the carotid arteries (p < 0.05) while maintaining the clinical target volume coverage. With conventional planning, median carotid V35, V50, and V63 were 100%, 100%, and 69.0%, respectively. With IMRT planning these decreased to 2%, 0%, and 0%, respectively (p < 0.01). Radiation planning and treatment times were similar for conventional radiotherapy and IMRT. Treatment results have been excellent thus far. CONCLUSIONS: Intensity modulated radiotherapy significantly reduced unnecessary radiation dose to the carotid arteries compared with conventional lateral fields while maintaining clinical target volume coverage. Further experience and longer follow-up will be required to demonstrate outcomes for cancer control and carotid artery effects. PMID- 19679407 TI - Influence of irradiated lung volumes on perioperative morbidity and mortality in patients after neoadjuvant radiochemotherapy for esophageal cancer. AB - PURPOSE: In some randomized trials, the treatment outcome of locally advanced esophageal cancer has been significantly improved by neoadjuvant radiochemotherapy (RCT). However, increased perioperative pulmonary toxicity in terms of acute respiratory distress syndrome (ARDS) has been linked to radiation exposure of the lungs. In our study we evaluated perioperative morbidity and mortality in patients with cancer Stages IIA-IVA treated with curative intent either with surgery alone (S) or with neoadjuvant RCT followed by surgery (RCTS). PATIENTS AND METHODS: Between 1996 and 2003, 55 patients received S, and 98 received RCTS. In the RCTS group, most patients received two cycles of 5 fluorouracil plus cisplatinum simultaneously with normofractionated radiotherapy (40Gy). Four weeks later they underwent surgery. Endpoints were the incidence of acute lung injury (ALI), ARDS, other postoperative complications, and mortality within 31 days. RESULTS: Between both groups there were no significant differences between the incidence and severity of ALI and ARDS (RCTS: 42.9%, 42.9%; S: 45.5%, 38.2%). Furthermore, there were no significant differences in the incidences of pneumonia, pleural effusion, and pneumothorax (RCTS 29.6% vs. S 16.4%, p = 0.07). Perioperative complication rates and mortality did not vary significantly (mortality after RCTS 5.1% vs. S 3.6%). A detailed analysis of 54 RCTS patients according to lung dose-volume histograms did not show any correlation between ARDS and pulmonary exposure. In univariate analysis, only respiratory comorbidity correlated with ARDS. CONCLUSION: Neoadjuvant cisplatinum and 5-fluorouracil-based RCT apparently has no detrimental impact on the postoperative course. PMID- 19679409 TI - [Amniocentesis and viral risk (hepatitis B, C virus and HIV)]. AB - Very few studies have properly addressed to the risk of fetal hepatitis B (HBV), hepatitis C (HCV) or human immunodeficiency virus (HIV) infection through amniocentesis. For HBV, this risk is low. However, knowledge of the maternal hepatitis B e antigen status is valuable in the counselling of risks associated with amniocentesis. For HCV, the risk is not well known but cannot be excluded. For HIV, it seems rational to propose a viral test before amniocentesis for patients with contamination's risk and to postpone the sampling in cases with positive results in order to obtain an undetectable HIV-1 RNA viral load. For these reasons, it can be useful to analyse for each virus the benefit of amniocentesis and the risk of mother-to-infant transmission, and to inform the patient. PMID- 19679408 TI - Locoregional tumor progression after radiation therapy influences overall survival in pediatric patients with neuroblastoma. AB - PURPOSE: There is renewed attention to primary site irradiation and local control for patients with high-risk neuroblastoma (NB). We conducted a retrospective review to identify factors that might predict for locoregional tumor control and its impact on overall survival. METHODS AND MATERIALS: Between July 2000 through August 2006, a total of 44 pediatric patients with NB received radiation therapy (RT) with curative intent using computed tomography (CT)-based treatment planning. The median age was 3.4 years and the median cumulative dose was 23.4 Gy. Overall survival and locoregional tumor control were measured from the start of RT to the date of death or event as determined by CT/magnetic resonance imaging/meta-iodobenzylguanidine. The influence of age at irradiation, gender, race, cumulative radiation dose, International Neuroblastoma Staging System stage, treatment protocol and resection status was determined with respect to locoregional tumor control. RESULTS: With a median follow-up of 34 months +/- 21 months, locoregional tumor progression was observed in 11 (25%) and was evenly divided between primary site and adjacent nodal/visceral site failure. The influence of locoregional control reached borderline statistical significance (p = 0.06). Age (p = 0.5), dose (p = 0.6), resection status (p = 0.7), and International Neuroblastoma Staging System stage (p = 0.08) did not influence overall survival. CONCLUSIONS: Overall survival in high-risk neuroblastoma is influenced by locoregional tumor control. Despite CT-based planning, progression in adjacent nodal/visceral sites appears to be common; this requires further investigation regarding target volume definitions, dose, and the effects of systemic therapy. PMID- 19679410 TI - The CB1 antagonist rimonabant (SR141716) blocks cue-induced reinstatement of cocaine seeking and other context and extinction phenomena predictive of relapse. AB - Cannabinoid CB1 antagonists decrease self-administration of palatable food and several abused drugs in animals and modulate extinction of conditioned fear responses. Less is known, however, about whether and how CB1 antagonists might modulate the extinction of appetitive behavior. Therefore, this study examined the effects of the CB1 receptor antagonist rimonabant (SR141716) during extinction of responding maintained either by cocaine or by palatable foods (corn oil or Ensure), as well as responding elicited by stimulus cues that had been paired with the presentation of cocaine (i.e., cue-induced reinstatement) or a prime (presentation of cocaine or food). The effect of rimonabant on high rate responding in water-deprived mice trained to self-administer water was also examined. In mice self-administering cocaine, rimonabant attenuated cue-induced reinstatement of cocaine self-administration, the initial burst of responding during cocaine extinction and responding during spontaneous recovery. In mice self-administering corn oil, rimonabant decreased responding during extinction and also attenuated responding that had been reinstated by a priming presentation of corn oil. Moreover, mice treated with rimonabant required fewer daily sessions to reach criterion for extinction of cocaine-maintained responding than vehicle treated mice. Also, rimonabant had no effect on the rate of operant responding in mice trained to respond for water under an FR5 schedule of reinforcement. Taken together, these data suggest that in addition to attenuating the primary reinforcing effects of both palatable foods and drugs of abuse, CB1 receptor antagonism can attenuate context and cue reactivity during extinction learning and potentially enhance extinction learning in this way. PMID- 19679412 TI - Out-of-season sperm cryopreserved in different media of the Amazonian freshwater fish pirapitinga (Piaractus brachypomus). AB - The pirapitinga (Piaractus brachypomus) is a freshwater fish that inhabits the Amazon and Orinoco River basins. The use of cryopreserved sperm has been considered to facilitate procedures during the artificial reproduction. The aim of the present study was to develop a freezing protocol for pirapitinga sperm collected outside the spawning season. Sperm samples were diluted in four freezing media prepared by a combination of two extenders (glucose and BTS Beltsville Thawing Solution) and two cryoprotectant agents (DMSO and methylglycol) loaded into 0.5-mL straws, frozen in a nitrogen-vapor shipping dewar (dry-shipper) and stored in liquid nitrogen at -196 degrees C. Post-thaw sperm motility was evaluated both subjectively using a light microscope and by a computer-assisted sperm analyzer (CASA). Curvilinear, average path and straight line velocities were also determined. There were no differences (P>0.05) in post thaw sperm motility between evaluations performed subjectively and using the CASA. Sperm samples cryopreserved in glucose-methylglycol yielded the greatest post-thaw sperm motility (81%) and fastest sperm velocities when compared to the samples frozen in the other three media (P<0.05). Out-of-season sperm cryopreserved in glucose and methylglycol under the conditions described above is of high quality and can therefore be used to facilitate artificial reproduction procedures, as only females will need handling for hormonal induction and gamete collection during the spawning season. Although the CASA system provides precise data on sperm motility, the subjective evaluation is practical and can be conducted by well-trained personnel at commercial fish farms as an acceptable evaluation of sperm quality. PMID- 19679414 TI - The evaluation of age-related histomorphometric variables in a cadaver sample of lower socioeconomic status: implications for estimating age at death. AB - Estimating age at death from adult skeletal remains is a daunting task for human osteologists. For this reason, the evaluation of micro-structural changes in bone with advancing age has become a popular method. However, factors such as nutrition, chronic disease, population group and sex have been suggested to influence the rate of bone turnover, and thus the use of histological methods in providing an accurate age at death has been questioned. The purpose of this study was to evaluate the accuracy and repeatability of 10 histomorphometric traits used to estimate age. The sample comprised of 146 dissection room cadavers of known sex, age and ancestry (105 males and 41 females). A 0.2cmx1.0cm sample was removed from the anterior surface of the mid-shaft of the femur (opposite the linea aspera), and slides were prepared according to standard methodology. The total osteon count (r=0.50), the percentage unremodelled bone (r=-0.50), the total number of non-Haversian canals (r=-0.50) and the average percentage of fragmental bone (r=0.55) had moderate correlations with age, while the total number of measurable osteons (r=0.43), the total number of osteonal fragments (r=0.40), the percentage of fragmentary bone (r=0.37) the average number of lamellae per osteon (r=0.29), the minimum diameter of the Haversian canals (r=0.14) and resorption spaces (r=0.11) had little to no relationship with age. Despite poor correlations with age, eight variables were shown to be highly repeatable (r=0.74-0.93). Moderate to low correlations with age may be attributed to these variables not being related to age in a progressive and predictable fashion, activity patterns or possible chronic disease in the sample. A databank needs to be compiled from larger samples from various populations in order to more holistically assess the relationship between these variables and age as well as other mitigating factors such as disease, nutrition and population group. PMID- 19679411 TI - Evaluation of prevalent phytocannabinoids in the acetic acid model of visceral nociception. AB - Considerable preclinical research has demonstrated the efficacy of Delta(9) tetrahydrocannabinol (Delta(9)-THC), the primary psychoactive constituent of Cannabis sativa, in a wide variety of animal models of pain, but few studies have examined other phytocannabinoids. Indeed, other plant-derived cannabinoids, including cannabidiol (CBD), cannabinol (CBN), and cannabichromene (CBC) elicit antinociceptive effects in some assays. In contrast, tetrahydrocannabivarin (THCV), another component of cannabis, antagonizes the pharmacological effects of Delta(9)-THC. These results suggest that various constituents of this plant may interact in a complex manner to modulate pain. The primary purpose of the present study was to assess the antinociceptive effects of these other prevalent phytocannabinoids in the acetic acid stretching test, a rodent visceral pain model. Of the cannabinoid compounds tested, Delta(9)-THC and CBN bound to the CB(1) receptor and produced antinociceptive effects. The CB(1) receptor antagonist, rimonabant, but not the CB(2) receptor antagonist, SR144528, blocked the antinociceptive effects of both compounds. Although THCV bound to the CB(1) receptor with similar affinity as Delta(9)-THC, it had no effects when administered alone, but antagonized the antinociceptive effects of Delta(9)-THC when both drugs were given in combination. Importantly, the antinociceptive effects of Delta(9)-THC and CBN occurred at lower doses than those necessary to produce locomotor suppression, suggesting motor dysfunction did not account for the decreases in acetic acid-induced abdominal stretching. These data raise the intriguing possibility that other constituents of cannabis can be used to modify the pharmacological effects of Delta(9)-THC by either eliciting antinociceptive effects (i.e., CBN) or antagonizing (i.e., THCV) the actions of Delta(9)-THC. PMID- 19679413 TI - Control of rat tail skin temperature regulation by estrogen receptor-beta selective ligand. AB - OBJECTIVE: To test the role of ERbeta in the control of estrogen-dependent thermoregulation in rats. METHODS: Test the ability of an ERbeta-selective ligand to suppress the elevation in basal rat tail skin temperature (TST) caused by ovariectomy (OVX). RESULTS: ERbeta-19 is a tetrahydrofluorenone ERbeta-selective ligand that displaces 0.1 nM estradiol from ERbeta with an IC50 of 1.8 nM compared to an IC50 of 141 nM for ERalpha. Like estradiol, it acts as an agonist on ERbeta-mediated transactivation and transrepression with 25- and 60-fold selectivity, respectively, over ERalpha-controlled transcription. Administration of estradiol to estrogen-depleted rats suppresses the ovariectomy-induced elevation of TST. Similar treatment of OVX rats with ERbeta-19 also results in suppression of elevated TST. However, in contrast to estradiol, ERbeta-19 does not suppress body weight, does not increase uterine weight, nor does it stimulate uterocalin biomarker expression which is under the control of ERalpha. Thus, the ERbeta-19 suppression of rat TST is mediated by ERbeta without eliciting the activity of ERalpha. CONCLUSION: Estrogen-sensitive thermoregulation in ovariectomized rats can be controlled by an ERbeta-selective ligand. PMID- 19679415 TI - The axonal damage marker tau protein in the cerebrospinal fluid is increased in patients with acute encephalopathy with biphasic seizures and late reduced diffusion. AB - Acute encephalopathy with biphasic seizures and late reduced diffusion (AESD) is a recently clinicoradiologically-established encephalopathy syndrome. In the present study, we examined the levels of cerebrospinal fluid (CSF) tau protein, a marker of axonal damage, in 11 patients with AESD. CSF tau levels were normal on day 1 and increased from day 3 of the disease between the initial and the secondary seizures. Magnetic resonance imaging (MRI) reveals reduced diffusion in the subcortical white matter during days 3-7. Two patients showed elevated tau protein prior to the diffusion abnormality of subcortical white matter on MRI. Levels of CSF neuron specific enolase (NSE), a neuronal marker, were elevated in only two out of seven patients with AESD, and CSF tau levels were also increased in these patients. Our results indicated that tau protein is a more sensitive marker than NSE and axonal damage causes the conspicuous MRI findings in AESD patients. A therapeutic strategy for axonal protection should be developed to prevent severe neurological impairment of AESD patients. PMID- 19679416 TI - [Future prospects in digestive endoscopy: wireless capsule endoscopy]. AB - Since the beginning of the millennium, the development of wireless capsule endoscopy has represented a major technological advance. The capsule is ingested by the patient and images are transmitted via several sensors positioned on the skin of the patient and downloaded in a computer system. The first applications were focused on the exploration of the small bowel which was previously considered as an obscure area for conventional endoscopy. Wireless capsule endoscopy of the small bowel is now an established technique with many acknowledged indications for the diagnosis of obscure bleeding, anemia of presumed digestive origin, Crohn's disease and small bowel tumors. Recently, thanks to technological progresses, novel capsules have been developed for specific segments of the gut namely the oesophagus and the colon. Recent data suggest that these new capsules could have potential applications for the diagnosis of oesophageal varices, Barrett's oesophagus and for the screening and/or surveillance of polyps of the colon. However, further studies are required before such strategies could be approved for clinical use or even replace conventional endoscopic modalities. In the long-term, progresses in signal processing as well as in the miniaturisation of sensors or markers may lead to a new generation of endoscopic robots. This technological breakthrough may ultimately result in new concepts and change current practice of digestive endoscopy. PMID- 19679417 TI - Contrast-enhanced MRA of the renal and aorto-iliac-femoral arteries: comparison of gadobenate dimeglumine and gadofosveset trisodium. AB - RATIONALE AND OBJECTIVES: Dedicated contrast agents are now available for contrast-enhanced magnetic resonance angiography (CE-MRA). This study retrospectively compares the safety and diagnostic performance data from Phase III regulatory trials performed to evaluate gadobenate dimeglumine (MultiHance((r))) and gadofosveset trisodium (Vasovist(r))) for renal and peripheral CE-MRA. MATERIALS AND METHODS: Similar examination and blinded assessment methodology was utilized in all studies to determine the safety and diagnostic performance of the agents for detection of significant (>50%) steno occlusive disease. Digital Subtraction Angiography (DSA) was used as the standard of truth. Diagnostic performance data (sensitivity, specificity, predictive values [PVs], and likelihood ratios [LRs]) were compared (Chi-square test). RESULTS: CE-MRA with gadobenate dimeglumine was more specific (92.4% vs. 80.5%, p < 0.0001) and accurate (83.6% vs. 77.1%, p = 0.022) than CE-MRA with gadofosveset in the detection of significant renal artery stenosis. The average sensitivity was higher for gadofosveset (74.4% vs. 67.3%, p = 0.011) in peripheral vessels although gadobenate dimeglumine was more specific (93.0% vs. 88.2%, p < 0.0001) with no difference in accuracy (86.6% vs. 86.3%, p = 0.66). PPVs were higher (p < 0.0001) for gadobenate dimeglumine in both vascular territories. Pre- to post test shifts in the probability of detecting significant disease were greater after gadobenate dimeglumine. Adverse events in the renal and peripheral studies were reported by 9.2% and 7.7% of patients after gadobenate dimeglumine compared with 30.3% and 22.1% of patients after gadofosveset. CONCLUSION: The diagnostic performance of CE-MRA for the detection of significant steno-occlusive disease is similar with gadofosveset and gadobenate dimeglumine although the rate of adverse events appears higher with gadofosveset. PMID- 19679418 TI - "Candidatus Paraholospora nucleivisitans", an intracellular bacterium in Paramecium sexaurelia shuttles between the cytoplasm and the nucleus of its host. AB - An intracellular bacterium was discovered in two isolates of Paramecium sexaurelia from an aquarium with tropical fish in Munster (Germany) and from a pond in the Wilhelma zoological-botanical garden, Stuttgart (Germany). The bacteria were regularly observed in the cytoplasm of the host, but on some occasions they were found in the macronucleus of the host cell. In these cases, only a few, if any, bacteria were observed remaining in the cytoplasm. The bacterium was not infectious to P. sexaurelia or other species of Paramecium and appeared to be an obligate intracellular bacterium, while bacteria-free host cells were completely viable. The fluorescence in situ hybridisation (FISH) and comparative 16SrDNA sequence analyses showed that the bacterium belonged to a new genus, and was most closely, yet quite distantly, related to Holospora obtusa. In spite of this relationship, the new bacteria differed from Holospora by at least two biological features. Whereas all Holospora species reside exclusively in the nuclei of various species of Paramecium and show a life cycle with a morphologically distinct infectious form, for the new bacterium no infectious form and no life cycle have been observed. For the new bacterium, the name Candidatus Paraholospora nucleivisitans is suggested. The host P. sexaurelia is usually known from tropical and subtropical areas and is not a species typically found in Germany and central Europe. Possibly, it had been taken to Germany with fish or plants from tropical or subtropical waters. Candidatus Paraholospora nucleivisitans may therefore be regarded as an intracellular neobacterium for Germany. PMID- 19679419 TI - Candidatus Monilibacter spp., common bulking filaments in activated sludge, are members of Cluster III Defluviicoccus. AB - Two alphaproteobacterial Neisser negative 'Nostocoida limicola' morphotypes differing slightly in their trichome diameter and filament regularity were dominant populations in the Bendigo, Victoria, Australia activated sludge community removing phosphorus (P). Neither responded to the FISH probes available for any of the other alphaproteobacterial 'N. limicola' morphotypes. Instead both fluoresced with the DF988 FISH probe designed originally to target alphaproteobacterial cluster II Defluviicoccus tetrad forming organisms. A 16S rRNA based clone library from this biomass revealed that the alphaproteobacterial clones grouped closely with Candidatus 'Monilibacter batavus' and Defluviicoccus clones in a cluster separate from the existing cluster I and II Defluviicoccus. When a FISH probe was designed against these, it only hybridized to the thinner and less abundant 'N. limicola' morphotype. Micromanipulation-RT-PCR was used to selectively recover the main 'N. limicola' morphotype and a FISH probe designed against the 16S rRNA clones generated from it showed only this filament fluoresced. From FISH based surveys, both 'N. limicola' variants occurred frequently in phosphorus removal activated sludge systems in Australia treating domestic waste. The data suggest that they represent two new strains of Candidatus 'Monilibacter', which on this evidence are filamentous members of the genus Defluviicoccus, a potential competitor for the polyphosphate accumulating organisms in these communities. PMID- 19679420 TI - On-chip assay for determining the inhibitory effects and modes of action of drugs against xanthine oxidase. AB - Xanthine oxidase (XO) is a key enzyme that can catalyze the conversion of xanthine to uric acid, causing various diseases in humans. We have developed a high-throughput chip-based assay that uses a photodiode array (PDA) microchip system to explore the inhibitory effects of drug analogs on XO. Inhibitory activities of cyclosporin A, aminoglutethimide, dithranol and naringenin against XO were assessed using this chip-based xanthine assay in the presence or absence of the antioxidant enzyme, superoxide dismutase (SOD). In addition, the mechanism of drug action was also disclosed by monitoring the combined effect of respective drug analogs and SOD on XO in the assay. The assessment was based on the red light absorption property of nitroblue tetrazolium (NBT) formazan, formed by free radical-mediated NBT reduction. Compared to naringenin (50 and 100 microM; a known XO inhibitor), cyclosporin A (5 and 10 microM) exhibited similar XO inhibitory activity, whereas dithranol (1 and 3 microM) and aminoglutethimide (2.5 and 5mM) showed minimum XO inhibition. Low standard deviation obtained during the assay demonstrates the preciseness and accuracy of the developed approach. Compared to the existing methods, the developed approach is advantageous due to its simplicity and compatibility with high-throughput screening procedures. Furthermore, this approach can be applied to the early phase of drug discovery screening to explore various drug analogs for their XO inhibitory activities. PMID- 19679421 TI - HAHA--nothing to laugh about. Measuring the immunogenicity (human anti-human antibody response) induced by humanized monoclonal antibodies applying ELISA and SPR technology. AB - Immunogenicity induced by passively applied proteins is a serious issue because it is directly related to the patient's safety. The out-come of an immune reaction to a therapeutic protein can range from transient appearance of antibodies without any clinical significance to severe life threatening conditions. Within this article, enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR) methodology to measure immunogenicity are compared and the pros and cons are discussed. PMID- 19679422 TI - What parents and carers think medical students should be learning about communication with children and families. AB - OBJECTIVE: Our objective was to dynamically engage with parents and carers of children treated in a large children's hospital as experts by experience, to find out what they thought medical students should be learning about communicating with children and families in order to inform our communication teaching. METHODS: We used focus groups to facilitate parents and carers in articulating the communication issues they felt were important for medical students to learn. These data were analysed, using qualitative and quantitative methods, to produce a survey for a Delphi consultation. The results of this stage were mapped onto the Calgary-Cambridge framework for the medical interview. RESULTS: There was considerable overlap of the data on the Calgary-Cambridge framework. There was, however, an emphasis by respondents on perceptual skills, self-awareness and partnership. Within the main tasks of the Calgary-Cambridge framework, new objectives emerged, including giving information in the right place, and the importance of the family in the multidisciplinary team. A new main task, Preparation, was highlighted. CONCLUSION: Active collaboration with parents and carers revealed areas for enhancement in our communication skill teaching, particularly with regard to relational and perceptual skills. PRACTICE IMPLICATIONS: Ways of including effective teaching of perceptual skills and the relational and humanistic aspects of communication must be explored in order to meet the needs of child patients and their families. PMID- 19679423 TI - Current evidence and clinical implications of aspirin resistance. AB - Atherothrombosis, characterized by atherosclerotic plaque rupture and subsequent occlusive or subocclusive thrombus formation is the primary cause of acute ischemic syndromes involving all vascular beds and accounts for more than one third of all deaths in the developed world. Platelet activation and aggregation constitute the most critical component in the pathophysiology of atherothrombotic disease. Aspirin is currently the most commonly used antiplatelet agent and one of the most frequently prescribed drugs, with as many as 30 million Americans on chronic aspirin regimens. Multiple well-designed prospective randomized clinical trials have demonstrated aspirin's efficacy in both primary and secondary prevention of a wide variety of entities that the atherothrombotic disease spectrum encompasses, such as cerebrovascular, coronary artery, and peripheral vascular disease. Despite its proven benefit, however, a growing body of evidence suggests that up to 70% of aspirin-takers may still be at risk for atherothrombotic complications due to resistance. Patients with laboratory confirmed aspirin resistance seem to have an almost fourfold increase in their risk for acute thrombotic episodes, which underlines the magnitude of the problem for the vascular specialist. In this article, we review the physiology of platelet activation and the role of aspirin as an antiplatelet agent; the various laboratory assays used in assessing aspirin effectiveness; and current data on aspirin resistance and its clinical implications in patients with cardiovascular disease. We also review the studies that explore this phenomenon in patients with peripheral arterial disease and discuss the optimal management options in aspirin resistant individuals. Suggestions are advanced for the direction of future trials evaluating aspirin resistance in patients with vascular disease. PMID- 19679425 TI - Secure fixation following EVAR with the Powerlink XL System in wide aortic necks: results of a prospective, multicenter trial. AB - PURPOSE: Endovascular stent graft repair of abdominal aortic aneurysms (AAA) with the Endologix Powerlink System (Endologix, Inc, Irvine, Calif) has been shown to be a safe and effective alternative to open surgery in patients having an aortic neck diameter of up to 26 mm. We assessed the safety and effectiveness of AAA repair in patients with wide aortic necks (up to 32 mm in diameter) using the Powerlink XL System. METHODS: Between September 2005 and June 2008, a prospective, multicenter, pivotal US Food and Drug Administration trial of the Powerlink XL System for endovascular aneurysm repair was conducted at 13 centers. Using a sizing algorithm based on computed tomography scan (CT)-based measurements, a total of 78 patients (N = 60 [pivotal trial]; N = 18 [continued access]) presenting with AAA and an infrarenal aortic neck up to 32 mm in diameter received a bifurcated stent graft via anatomical fixation at the aortoiliac bifurcation and proximal sealing with a Powerlink XL infrarenal proximal extension stent graft. Postoperatively, results were assessed with contrast-enhanced CT scans and abdominal x-rays at one, six, and 12 months, with continued annual follow-up to five years. RESULTS: Predominantly male (91%), patients presented at a mean age of 73 +/- 8.6 years with mean maximum aortic neck and AAA diameters of 31 +/- 1.9 mm (range, 25 to 32 mm) and 5.7 +/- 1.0 cm (range, 4.3 to 10 cm), respectively. Challenging infrarenal aortic neck anatomy, defined as the presence of severe thrombus and/or reverse taper, was present in 85% of patients. Technical success was achieved in 98.7% of patients, with one patient requiring femoral-femoral bypass intraoperatively. Aneurysm exclusion was achieved in 100% of patients over a mean procedure time of 129 +/- 66 minutes. Patients were discharged at a mean of 2.2 days postoperatively. At the one-month CT scan, the independent core lab identified a Type II endoleak in 13 patients, distal Type I and Type II endoleak in one patient, and unknown endoleak in three patients. At 30 days, there were no deaths, conversions, ruptures, or migrations. Through one year follow-up, Type II endoleak predominated (9/10 patients with endoleak), with one proximal Type I and no Type III, IV, or unknown endoleak; no conversions, ruptures, or migrations have been observed. The one-year all-cause mortality rate was 6.4%, with 100% freedom from aneurysm-related mortality. Secondary procedures were performed within one year in five patients (6.4%) for treatment of proximal Type I endoleak (n = 2), proximal Type I/Type II endoleak (n = 1), and distal Type I endoleak (n = 2). Reduced or stable aneurysm sac diameter at one year is observed in 96% of patients. CONCLUSIONS: The combination of an anatomically-fixed Powerlink bifurcated stent graft and a Powerlink XL infrarenal proximal extension appears safe and effectively excludes aneurysms in patients with wide aortic necks. These results suggest that fixation at the aortic bifurcation can provide secure fixation for patients with large diameter diseased proximal aortic necks. PMID- 19679426 TI - Intraoperative salvage of a renal artery occlusion during fenestrated stent grafting. AB - A 79-year-old man with a 6-cm juxtarenal abdominal aortic aneurysm was treated by endovascular means with a fenestrated stent graft. The completion angiogram revealed a left renal artery occlusion. A retroperitoneal surgical approach allowed for retrograde catheterization of the occluded covered stent through the left renal artery. The covered stent was reopened by balloon angioplasty. After 2 months, the left renal artery was patent and renal function normal. At 6 months, both renal arteries were fully open on duplex imaging. The open retroperitoneal approach with retrograde catheterization is a bailout technique to avoid loss of a kidney in fenestrated stent grafting. PMID- 19679424 TI - In vivo human lower extremity saphenous vein bypass grafts manifest flow mediated vasodilation. AB - OBJECTIVE: As in arteries, venous endothelium modulates vessel homeostasis and tone. The effect of an arterialized environment on venous endothelial function remains poorly understood. In particular, regulation of saphenous vein graft (SVG) blood flow and lumen caliber remains undefined. We hypothesized that mature SVGs would exhibit endothelium-dependent, flow-mediated vasodilation (FMD). We further hypothesized that endothelium-derived nitric oxide (NO) was an important mediator. METHODS: Patients with femoral to popliteal artery SVGs that had maintained primary patency and were at least 1 year from surgery were enrolled. High-resolution, B-mode ultrasound scans were used to measure vein graft diameter before and 1 minute after reactive hyperemia (RH) to determine FMD. RH was created through application of 220 mm Hg to the calf for 5 minutes with a sphygmomanometric cuff. After a 10-minute recovery period, nitroglycerin mediated, endothelium-independent vasodilation was measured 3 minutes after administration of nitroglycerin 0.4 mg sublingually. Brachial artery FMD was determined by validated techniques. L-N(G)monomethyl arginine (L-NMMA; 1 mg/kg infusion over 10 minutes) was used in a subset of patients (n = 6) to competitively inhibit endothelial NO synthase. RESULTS: Nineteen subjects were enrolled. The median age of the SVGs was 34.6 (21.0-49.7) months. SVG flow mediated, endothelium-dependent vasodilation was measured at 5.28% +/- 3.1% mean change in lumen diameter (range, 1.99%-9.36%; P < .0001 for diameter change). Nitroglycerin-mediated vasodilation was 3.7% +/- 1.0%, (range, 16%-10.04%; P < .005). Intravenous administration of L-NMMA abolished SVG FMD (5.7 +/- 1.4% before L-NMMA vs 0.01 +/- 0.01% during L-NMMA infusion; P = .0088) and attenuated brachial artery FMD (7.54% +/- 1.0% vs 5.7 +/- 1.4; P = .05). CONCLUSION: SVGs manifest flow-mediated, endothelium-dependent, and nitroglycerin-mediated endothelium-independent vasodilation. Vein graft endothelium-dependent FMD is likely mediated by NO. Further investigation will be required to determine the role of endothelial function in vein graft patency. PMID- 19679427 TI - Angiogenic properties of sustained release platelet-rich plasma: characterization in-vitro and in the ischemic hind limb of the mouse. AB - BACKGROUND: While single growth factor has limitation to induce optimal neovascularization, platelet-rich plasma (PRP) is an autologous reserver of various growth factors. However, little is known about the mechanism of PRP related neovascularization.The objective of this investigation was to characterize the angiogenic and growth factor content of PRP and to determine, in vitro, its effect on endothelial cell proliferation. Additionally, this experiment sought to determine the effectiveness of different compositions of PRP (solution versus sustained release) on perfusion and neovascularization in a murine model of hind limb ischemia. METHODS: Different growth factors were measured by enzyme-linked immunosorbent assay (ELISA). In vivo study, we used gelatin hydrogel as a sustained release carrier for growth factors in PRP. We induced hind limb ischemia by excising right femoral artery in wild type C57BL6 mice. After surgery, mice were randomly assigned to four experimental groups; control (C), 100 muL of sustained release form of platelet-poor plasma (PPP), 100 muL of solution form of PRP (PRP-sol), 100 muL of sustained release form of PRP (PRP-sr); each formulation was administered via an intramuscular injection to the ischemic hind limb. Endpoint evaluations were blood perfusion by laser Doppler perfusion image, vascular density by anti Von Willebrand factor (vWF), and mature vessel density by anti smooth muscle actin (SMA) antibody. Green fluorescent protein (GFP+) transgenic mice were generated by transplantation of bone marrow derived mononuclear cells to wild type C57BL6 mice, and finally CD34+ cell in the ischemic site of transgenic mice was detected by staining with anti-CD34 antibody. RESULTS: In vitro study showed that PRP containing different growth factors induces endothelial cell proliferation and capillary tube formation. In vivo study demonstrated that sustained release of PRP increased perfusion of ischemic tissue as measured by laser Doppler perfusion imaging (LDPI) (57 +/- 12, 56 +/- 9, 72 +/- 7, 98 +/- 4 for groups C, PPP, PRP-sol, and PRP-sr, respectively; P < .05); capillary density (151 +/- 16, 158 +/- 12, 189 +/- 39, 276 +/- 39 for groups C, PPP, PRP-sol, and PRP-sr, respectively, P < .05) and mature vessel density (28 +/- 2, 31 +/- 3, 52 +/- 10, 85 +/- 13 for groups C, PPP, PRP-sol, and PRP-sr, respectively, P < .05) . Sustained release PRP also increases CD34+ cells in the ischemic site of transgenic mice (6 +/- 3 vs 18 +/- 5/mm(2) for groups control and PRP-sr respectively, P < .05). CONCLUSION: Sustained release of PRP containing potent angiogenic growth factors restores blood perfusion presumably by stimulating angiogenesis, arteriogenesis, as well as vasculogenesis in the mouse hind limb ischemia. CLINICAL RELEVANCE: PRP is a natural reserver of various growth factors that can be collected autologously and is costeffective. Thus for clinical use, no special considerations concerning antibody formation and infection risk are needed.Some clinical devices to automatically prepare PRP are available at present. PRP are consistently being used clinically inthe department of orthopedics and plastic surgery (oral, maxillary facial) for a long time. On the basis of researchevidence, some publications have reported positive results in either bone or soft tissue healing. However, some researchconcludes that there is no or little benefit from PRP. This is likely due to faster degradation of growth factors in PRP sincesome authors suggest using sustained release form of PRP to deliver optimal effect of PRP. Gelatin hydrogel is also beingused clinically as a slow, sustained release of carrier for growth factors in our center recently. PMID- 19679428 TI - [Availability of antidotes in emergency]. PMID- 19679429 TI - [Multiple pulmonary embolism of cement after vertebroplasty]. PMID- 19679430 TI - [Internet-based continuing medical education: as effective as live continuing medical education]. AB - E-learning consists in using new multimedia and Internet technologies to improve the quality of learning activities by facilitating access to resources and services, as well as exchanges and remote collaboration. The Internet is used for adult education in most professional domains, but its use for continuing medical education is less developed. Advantages are observed for teachers (e.g., permanent updating, interactive links, illustrations, archiving, and collective intelligence) and for the learners (e.g., accessibility, autonomy, flexibility, and adaptable pace). Research and meta-analyses have shown that e-CME is as effective as live events for immediate and retained learning. English-language educational medical websites that grant CME credits are numerous; few such French language sites can currently grant credits. Accreditation of websites for CME, in its infancy in Europe, is common in North America. PMID- 19679431 TI - Cementless total hip arthroplasty specifically designed for asians: clinical and radiologic results at a mean of 10 years. AB - It is known that the proximal femoral osteometry is different between Asians and whites. An uncemented femoral stem was therefore designed by us specifically for Asians, and we now report the medium-term clinical results. Results of a proximally porous-coated femoral component anatomically matched for Asians were analyzed in a consecutive cohort of 48 hips. The mean follow-up was 10.3 years. The mean Harris hip score was 39.4 preoperatively and 94.2 at last follow-up (P < .001). All stems had bone ingrowth. Stem survival was 100% up to 14 years using revision as the end point. Eight acetabular component revisions were performed for bearing surface failures and osteolysis. This femoral stem, specifically designed for better fit in an Asian population, had excellent clinical and radiologic outcome. PMID- 19679432 TI - Does ischial screw fixation improve mechanical stability in revision total hip arthroplasty? AB - Ischial screw fixation, albeit technically challenging, is postulated to provide additional mechanical stability in revision total hip arthroplasty (THA). Hemipelvis specimens were prepared to simulate revision THA, and an acetabular component with supplemental screw fixation was implanted. Three configurations were tested: 2 dome screws alone, 2 dome screws plus an additional screw within the dome, and 2 dome screws plus an ischial screw. Force displacement data were acquired during mechanical testing. An increase in mechanical stability was observed in acetabular components with supplemental screw fixation into either the posterior column or ischium (P<=.031) compared to isolated dome fixation. In addition, supplemental ischial screw fixation may provide a modest advantage over a screw placed posteroinferiorly within the acetabular dome during revision THA. PMID- 19679433 TI - Hip and knee arthroplasty in hypophosphatemic rickets. AB - There is very limited published information about the technical aspects and durability of lower extremity arthroplasty in hypophosphatemic rickets. Between 1972 and 2006, 8 total hip arthroplasties (THAs) and 6 total knee arthroplasties (TKAs) were performed in 8 patients with degenerative arthritis and bone deformity secondary to hypophosphatemic rickets. Two hips required osteotomies at the time of arthroplasty, and 1 TKA patient required postoperative osteotomies. Specialized implants were required in 3 hips and 1 knee. At average follow-up of 7 years, mean Harris hip scores improved to 21 points, and mean Knee Society pain and function scores improved to 48 points and 27 points, respectively. One cemented THA failed due to femoral aseptic loosening at 13 years postoperatively; all other implants remained well fixed. Lower extremity arthroplasty is effective and durable for patients with arthritis associated with hypophosphatemic rickets, but corrective osteotomies and use of special implants should be anticipated with more severe deformities. PMID- 19679435 TI - Evaluation of phase stability in zirconia femoral heads from different manufacturers after in vitro testing or in vivo retrieval. AB - Yttria-stabilized zirconia femoral heads from 3 different manufacturers were tested in vitro with respect to their phase stability and compared with retrieved zirconia heads. The monoclinic content on the surface of unused heads was analyzed by confocal Raman spectroscopy after exposure for increasing times to moist atmosphere. The increase in monoclinic content was then plotted as a function of geometric location on the head surface of the head and compared with that measured at similar locations after in vivo exposure. Profiles of residual stress associated to polymorphic transformation were also measured from the collected Raman spectra. A striking finding was that, in some samples, polymorphic transformation occurred since the very early stage of the environmental exposure even if those samples belonged to new-generation products. PMID- 19679434 TI - Does aspirin have a role in venous thromboembolism prophylaxis in total knee arthroplasty patients? AB - The objectives of this study were to compare the risk of venous thromboembolism (VTE), bleeding, surgical site infection, and mortality in patients receiving aspirin or guideline-approved VTE prophylactic therapies (warfarin, low-molecular weight heparins, synthetic pentasaccharides) in total knee arthroplasty (TKA). We analyzed clinical and administrative data from 93,840 patients who underwent primary TKA at 307 US hospitals over a 24-month period. Fifty-one thousand nine hundred twenty-three (55%) patients received warfarin, 37,198 (40%) received injectable agents, and 4719 (5%) received aspirin. After adjustment for patient and hospital factors, patients who received aspirin VTE prophylaxis (VTEP) had lower odds for thromboembolism compared to warfarin patients but with similar odds compared with injectable VTEP; there were no differences in risk of bleeding, infection, or mortality after adjustment. Our results suggest that aspirin, when used in conjunction with other clinical care protocols, may be effective VTEP for certain TKA patients. PMID- 19679436 TI - Lateral epicondylar osteotomy using computer navigation in total knee arthroplasty for rigid valgus deformities. AB - Extensive release of posterolateral structures may be required to correct rigid and severe valgus deformities during total knee arthroplasty. Current techniques are technically difficult, may not accurately restore soft tissue balance, and are associated with postoperative complications. Computer navigation while performing lateral femoral epicondylar osteotomy allows precise, controlled, quantitative lengthening of lateral structures and restoration of optimum soft tissue balance and alignment. PMID- 19679437 TI - Predicting short-term outcome of primary total hip arthroplasty:a prospective multivariate regression analysis of 12 independent factors. AB - The purpose of this study was to investigate factors affecting the short-term outcome of primary total hip arthroplasty (THA) and develop a multivariate regression equation to predict the short-term outcome of primary THA. Prospectively collected data for 101 primary THAs performed at a single institution were used in this study. Twelve independent variables were analyzed via correlation and multivariate regression analyses. Correlation analyses showed that three variables significantly influenced Western Ontario and McMaster Universities Osteoarthritis (WOMAC) physical function (PF) score at minimum follow-up of 1 year: preoperative WOMAC PF score (P < .0001), sex (GN, P = .0159), and the presence of preoperative comorbidities (CMB, P = .0246). Multivariate regression analysis yielded the following equation: Outcome = PF0.45 - GN9 + CMB8 + 62, which can be used to predict the general short-term outcome of primary THA. PMID- 19679438 TI - The estimated magnitude and direct hospital costs of prosthetic joint infections in the United States, 1997 to 2004. AB - To estimate the number and cost of prosthetic joint infection hospitalizations in civilian US hospitals, we analyzed the 1997 to 2004 National Hospital Discharge Survey for the 996.66 International Classification of Diseases discharge code (infection or inflammatory reaction secondary to internal joint prosthesis). The annual number of such hospitalizations averaged 17 589 from 1997 to 2000 and 29 225 from 2001 to 2004. The annual adjusted diagnostic-related group cost for such infection increased from $195 million to $283 million (1997-2004). The mean diagnostic-related group reimbursement ($9034 per hospitalization) did not vary over time or by comorbidity. The nearly doubled number of prosthetic joint infection-related hospitalizations may have been caused by an increased implant rate, changes in patient population, implant procedures, or causative organisms. PMID- 19679439 TI - Is minimally invasive surgery-total knee arthroplasty truly less invasive than standard total knee arthroplasty? A quantitative evaluation. AB - With recent technical advancements, the number of operative manipulations in the knee joint by minimally invasive surgery-total knee arthroplasty (MIS-TKA) is now considered to be the same as that using standard TKA (S-TKA). The question still remains, however, if MIS-TKA improves recovery compared to S-TKA. We compared MIS TKA and S-TKA patients' physical activity as measured by an accelerometer. Physical activity expressed as cumulative acceleration was significantly higher in the MIS-TKA than in the S-TKA group on postoperative days (POD1, 2, 3, 4, 5, 10, 11) (P < .05). The recovery time, defined as the number of days required to achieve cumulative acceleration of 80% of the preoperative level, was significantly shorter (P < .05) in the MIS-TKA (3.0 +/- 3.3 days) group than in the S-TKA (7.0 +/- 3.5 days) group. Minimally invasive surgery-total knee arthroplasty appears to allow an earlier recovery after the operation than S-TKA. Less invasion to muscle during the surgery appears to contribute to shorter convalescence. PMID- 19679440 TI - Results of unicompartmental knee arthroplasty with cemented, fixed-bearing prosthesis using minimally invasive surgery. AB - The aim of this study was to evaluate the intermediate-term survivorship of cemented fixed-bearing unicompartmental knee arthroplasty (UKA). One hundred twenty-eight consecutive UKAs in 98 patients were performed by a single surgeon with "Allegretto" UKA prostheses. Six patients died and 5 were lost to follow-up, leaving 87 patients (116 knees) with an average follow-up of 5.7 years (range, 3 8 years). At the latest follow-up, the survival rate was 92.2%; average clinical and functional knee society scores improved from 50 and 64 points preoperatively to 89 and 88 points, respectively; and average total knee score was 171 (range, 94-200). Radiographic follow-up demonstrated average alignment of 3 degrees valgus (range, 6 degrees varus to 7 degrees valgus). Ninety-two percent of the patients indicated satisfaction with the procedure. Nine knees failed requiring revision at an average of 40.4 months. We are encouraged with the satisfactory results, considering the broader spectrum of selected patients. PMID- 19679441 TI - Adjustable cutting blocks for computer-navigated total knee arthroplasty: a cadaver study. AB - Computer-navigation in total knee arthroplasty has been reported to increase accuracy but also procedure duration. We compared surgical time and precision using a novel adjustable cutting block vs freehand navigation with conventional blocks on 12 bilateral cadaver tibiae. The mean time required was significantly less to position the adjustable block than the conventional block (2 minutes 10 seconds vs 6 minutes 35 seconds, P = .006). Guide positioning precision (standard deviation) for the adjustable block vs conventional block was as follows: varus/valgus, 0.24 degrees vs 1.16 degrees (P = .015); posterior slope, 0.35 degrees vs 0.74 degrees (P = .13); and cut height, 0.37 vs 1.41 mm (P = .010). There were no significant differences in the final bone cut accuracy between the 2 groups. The use of adjustable cutting blocks simplifies navigated procedures and may reduce the time required to perform a navigated total knee arthroplasty. PMID- 19679443 TI - Compression strength and porosity of single-antibiotic cement vacuum-mixed with vancomycin. AB - We evaluated the ultimate compression strength (UCS), porosity, and fracture surface roughness of 2 commercially available single-antibiotic bone cements vacuum-mixed with additional amounts of vancomycin (2, 4, 6, and 8 g). At least 8 g could be added to Palacos R + 0.5 g gentamicin (UCS = 75.04 +/- 6.64 MPa) and no more than 6 g to Simplex P + 1 g tobramycin (UCS = 78.93 +/- 4.98 MPa) to maintain a UCS above the International Organization for Standardization minimum standard (70 MPa). Increasing vancomycin concentration correlated with a decrease in porosity but showed a trend towards greater fracture surface roughness. PMID- 19679442 TI - Transvaginal emergence of a broken cup marker ring 21 years after total hip arthroplasty. AB - Broken cups, wires, and cable systems have the potential to cause complications, and they can migrate into joints, abdominal and thoracic organs. We describe a case of migration and transvaginal emergence of a marker wire from a polyethylene cup. PMID- 19679445 TI - Two forms of feedback inhibition determine the dynamical state of a small hippocampal network. AB - Pyramidal cells in the hippocampus are part of a small neuronal network that performs computations on external input. The network consists of principal cells and various forms of feedback inhibition. Experimental evidence indicates at least two functionally distinct inhibitory feedback loops in the CA3 area of the hippocampus: (1) a loop in which O-LM interneurons project to the distal dendrites of pyramidal cells with synapses that have slow kinetics, and (2) a loop in which basket interneurons project to the somata of pyramidal cells with synapses that have fast kinetics. There is an interconnection between the two loops in the form of O-LM to basket interneuron inhibition and the configuration is further complicated by the presence of distinct propagation delays and short term facilitation and depression of certain synapses in the two basic loops. In this study we investigated the consequences of various configurations of the circuit and modulations of the components of inhibition for the computation that the network can perform on its input. Gaussian noise was used as the input to the dendrite of the pyramidal cell and evoked two types of events: spikes or bursts. The event-triggered average (ETA) and the event-triggered covariance (ETC) were determined and the inter-event-intervals between spikes and bursts were analyzed. The ETA and ETC on the pyramidal cell show that this model behaves in first approximation as an activity integrator: with sufficient positive input, bursts as well as spikes are evoked. Which of the two is determined by the input just after the (first) spike: positive input results in a burst; negative input results in a spike. Stronger feedback inhibition, in the slow as well as in the fast loop, increases the event rate of the pyramidal cell. For a single input and large propagation delays, the interaction between the two feedback loops is not of great importance. The consequences of the presence of the slow and/or fast feedback inhibitory loop, with or without facilitation and depression, were analyzed in relation to synapse strength. Facilitation and depression are most relevant when their recovery time constant is of the same order as the mean inter event interval. Short-term depression can stop activity in the fast loop after several fast spikes and can switch the network to a different state, thus functioning as a kind of 'brake' on the fast inhibitory feedback loop. Thus inhibition and the details of the microcircuit organization play an important role in the information processing of the small neuronal circuit. PMID- 19679444 TI - Clinical results of the wear performance of cross-linked polyethylene in total hip arthroplasty: prospective randomized trial. AB - To investigate the clinical results of cross-linked polyethylene (CLPE) and to compare the CLPE wear against zirconia and stainless steel heads, we studied the radiographic wear after a minimum 3-year follow-up in total hip arthroplasty (THA). Ninety-four hips were randomly implanted with a 22.225-mm head cemented THA-the group of non-CLPE against zirconia and CLPE against 2 different zirconias and stainless steel. The linear wear rate was significantly lower in the group of CLPE against zirconia (0.067, 0.059 mm/y) and against stainless steel (0.068 mm/y) compared with non-CLPE against zirconia (0.170 mm/y). In the short-term results, the wear performance of CLPE against zirconia was superior to that of non-CLPE; however, it did not show a better wear rate than CLPE against stainless steel. Furthermore, long-term investigations will be necessary for understanding CLPE wear in vivo. PMID- 19679446 TI - Response shift and outcome assessment in orthopedic surgery: is there a difference between complete and partial treatment? PMID- 19679447 TI - Autoimmune myocarditis: past, present and future. AB - Heart failure has become an increasingly prevalent disorder with considerable morbidity and mortality. While many causal mechanisms such as inherited cardiomyopathies, ischemic cardiomyopathy or muscular overload are easily identified in clinical practice, the molecular mechanisms that determine the progression of heart failure or ventricular remodelling are largely unknown. Autoimmune responses and inflammation are involved in the pathogenesis of many cardiovascular diseases. There is compelling evidence that inflammatory mechanisms may contribute to progressive heart failure. Thus, myocardial infiltration of lymphocytes and mononuclear cells, increased expression of pro inflammatory chemokines and cytokines and circulating autoantibodies are frequently observed in myocarditis and dilated cardiomyopathy. In this review we give an overview on myocarditis and describe why diagnosis and treatment of myocarditis in the clinic can be difficult. We present current animal models and describe possible experimental approaches to improve diagnosis. Finally, we give an outlook on possible drug targets by describing the latest findings in the animal models focussing on chemokines and cytokines, T cell responses and interactions, tolerance induction and the development of autoantibodies. PMID- 19679448 TI - Preliminary study of indirect CT lymphography-guided sentinel lymph node biopsy in a tongue VX2 carcinoma model. AB - The feasibility of using indirect CT lymphography (CT-LG) to guide sentinel lymph node (SLN) biopsy in a rabbit model of tongue VX2 carcinoma with cervical lymph node metastasis was studied. Tongue VX2 carcinoma with cervical lymph node metastasis was induced in 19 rabbits by injecting VX2 carcinoma suspension into the tongue submucosa. SLN biopsy was performed under the guidance of indirect CT LG. SLN identification was performed by indirect CT-LG combined with blue dye injection in 2 rabbits. Tongue SLNs were identified preoperatively by indirect CT LG and blue-stained SLNs were visualized intraoperatively. Only one SLN was enhanced in each side of the neck, lateral to the larynx-tracheal region. CT attenuation values of the enhanced SLNs were 782.4+/-46.6, 443.1+/-68.5, 180.3+/ 20.6 and 80.5+/-10.7 HU at 1, 5, 15 and 20 min after contrast injection. Overall, ipsilateral SLN identification rate was 97.4% and contralateral SLN identification rate was 100%. Ipsilateral SLN metastasis was verified in all rabbits (100%), bilateral SLN metastasis occurred in 8 rabbits (42%), and micrometastasis was found in 3 rabbits (16%). Indirect CT-LG may be useful for guiding SLN biopsies in tongue cancer. Combining indirect CT-LG with blue dye injection may improve preoperative and intraoperative SLN identification. PMID- 19679449 TI - Antiepileptic drugs in epilepsy and other disorders--a population-based study of prescriptions. AB - PURPOSE: The aim of the study was to quantify the use of antiepileptic drugs (AEDs) in epilepsy and other indications in a nation-wide population using a prescription database. MATERIALS AND METHODS: Prescription data of AEDs were collected from the Norwegian Prescription Database for the period 2004-2007, including 5.1 million prescriptions from 144,653 patients, all having at least one prescribed and dispensed AED. Diagnosis-related reimbursement codes were used as indicators for clinical use. RESULTS: Of the AEDs used, 71% was in epilepsy, 15% in psychiatry, 13% in neuropathic pain, and <1% in migraine, trigeminus neuralgia and cancer adjuvance. The use in epilepsy increased steadily from 7.0 to 7.5 DDDs/1000 inhabitants/day (7%) from 2004 to 2007. The use of AEDs in other indications increased considerably more from 2004 to 2007: Neuropathic pain to 1.40 DDDs/1000 inhabitants/day (360%), psychiatry to 1.59 (200%), and migraine to 0.005 (642%). The prevalence of AED users increased and in 2007 it was 0.97% in epilepsy, 0.8% in neuropathic pain, 0.33% in psychiatry and 0.001% in migraine. In 2007, 49% of the AEDs used in epilepsy were a new drug, carbamazepine, lamotrigine and valproate being most frequently used. New AEDs were used in 96% in migraine prophylaxis, 94% in neuropathic pain, and 64% in psychiatry. CONCLUSIONS: Based on a nation-wide prescription database, this study quantifies the wide and increasing use of AEDs, in particular the newer drugs. PMID- 19679450 TI - Effects of bilateral repetitive transcranial magnetic stimulation on treatment resistant auditory-verbal hallucinations in schizophrenia: a randomized controlled trial. AB - BACKGROUND: Neuroimaging findings implicate bilateral superior temporal regions in the genesis of auditory-verbal hallucinations (AVH). This study aimed to investigate whether 1 Hz repetitive transcranial magnetic stimulation (rTMS) of the bilateral temporo-parietal region would lead to increased effectiveness in the management of AVH, compared to left rTMS or placebo. METHODS: 38 patients with schizophrenia (DSM-IV) and medication-resistant AVH were randomly assigned to 1 Hz rTMS treatment of the left temporo-parietal region, bilateral temporo parietal regions, or placebo. Stimulation was conducted over 6 days, twice daily for 20 min, at 90% of the motor threshold. Effect measures included the Auditory Hallucination Rating Scale (AHRS), Positive and Negative Affect Scale (PANAS), and a score for hallucination severity obtained from the Positive and Negative Syndrome Scale (PANSS). RESULTS: All groups showed some improvement on the total AHRS. Hallucination frequency was significantly reduced in the left rTMS group only. The bilateral rTMS group demonstrated the most remarkable reduction in self reported affective responsiveness to AVH. A modest, but significant decrease on the PANSS hallucination item was observed in the combined rTMS treatment group, whereas no change occurred in the placebo group. The left rTMS group showed a significant reduction on the general psychopathology subscale. CONCLUSION: Compared to bilateral or sham stimulation, rTMS of the left temporo-parietal region appears most effective in reducing auditory hallucinations, and additionally may have an effect on general psychopathology. Placebo effects should however not be ruled out, since sham stimulation also led to improvement on a number of AVH parameters. PMID- 19679451 TI - Clozapine and quetiapine acutely reduce glucagon-like peptide-1 production and increase glucagon release in obese rats: implications for glucose metabolism and food choice behaviour. AB - OBJECTIVE: Second generation antipsychotic drug (SGA) treatment is associated with detrimental effects on glucose metabolism which is often attributed to the development of obesity and insulin resistance. However, we have recently demonstrated that clozapine and quetiapine also have direct effects of glucose metabolism in animals. This study compares clozapine and quetiapine and investigates the effects of these on the development of obesity and the direct effects of these drugs on glucose metabolism compared with those caused by the obesity per se. RESEARCH DESIGN AND METHODS: Three groups of male Sprague-Dawley rats were fed a high fat/high sugar diet to induce obesity while another three groups were fed a chow diet. One group on each diet was injected daily with vehicle, clozapine or quetiapine and effects on glucose metabolism were monitored. RESULTS: Clozapine and quetiapine treatment did not directly cause obesity or potentiate diet induced obesity but did induce a preference for the high fat/high sugar diet. Neither drug caused a impairment in insulin tolerance over that caused by obesity but both drugs acutely induced impairments in glucose tolerance that were additive with the effects induced by the diet induced obesity. Both drugs caused increases in glucagon levels and a suppression of GLP 1. We investigated two strategies for restoring GLP-1 signalling. The DPP-IV inhibitor sitagliptin only partially restored GLP-1 levels and did not overcome the deleterious effects on glucose tolerance whereas the GLP-1 receptor agonist exendin-4 normalised both glucagon levels and glucose metabolism. CONCLUSIONS: Our findings indicate that the clozapine and quetiapine induced impairments in glucose tolerance in rats are independent of insulin resistance caused by obesity and that these defects are linked with a suppression of GLP-1 levels. These studies suggest the need to perform follow up studies in humans to determine whether clozapine and quetiapine induce acute derangements in glucose metabolism and whether GLP-1 replacement therapy might be the most appropriate therapeutic strategy for treating derangements in glucose metabolism in subjects taking these drugs. PMID- 19679452 TI - Olopatadine hydrochloride inhibits scratching behavior induced by a proteinase activated receptor 2 agonist in mice. PMID- 19679453 TI - Improving mechanical and biological properties of macroporous HA scaffolds through composite coatings. AB - Interconnected porous hydroxyapatite (HA) scaffolds are widely used for bone repair and replacement, owing to their ability to support the adhesion, transfer, proliferation and differentiation of cells. In the present study, the polymer impregnation approach was adopted to produce porous HA scaffolds with three dimensional (3D) porous structures. These scaffolds have an advantage of highly interconnected porosity (approximately 85%) but a drawback of poor mechanical strength. Therefore, the as-prepared HA scaffolds were lined with composite polymer coatings in order to improve the mechanical properties and retain its good bioactivity and biocompatibility at the same time. The composite coatings were based on poly(D,L-lactide) (PDLLA) polymer solutions, and contained single component or combination of HA, calcium sulfate (CS) and chondroitin sulfate (ChS) powders. The effects of composite coatings on scaffold porosity, microstructure, mechanical property, in vitro mineralizing behavior, and cell attachment of the resultant scaffolds were investigated. The results showed that the scaffolds with composite coatings resulted in significant improvement in both mechanical and biological properties while retaining the 3D interconnected porous structure. The in vitro mineralizing behaviors were mainly related to the compositions of CS and ChS powders in the composite coatings. Excellent cell attachments were observed on the pure HA scaffold as well as the three types of composite scaffolds. These composite scaffolds with improved mechanical properties and bioactivities are promising bone substitutes in tissue engineering fields. PMID- 19679454 TI - Accessory extrahepatic arteries: Blood supply of a human liver by three arteries A case report with brief literature review. AB - The coexistence of three hepatic arteries - accessory left hepatic artery (aLHA), proper hepatic artery (pHA), and accessory right hepatic artery (aRHA) - was demonstrated during a routine dissection of an 85-year-old male cadaver. The aLHA arose from the left gastric artery and ran in the hepatogastric ligament, whereas the aRHA took its origin from the superior mesenteric artery and ran in the hepatoduodenal ligament. The anatomy of the origins and the course of the arteries in the liver are described. Knowledge of these variations is of importance for surgical and radiological procedures in the upper abdominal region, in order to avoid complications during invasive treatment. The embryological background of the variations is discussed. PMID- 19679455 TI - Pharmacokinetic interaction studies of tanshinones with tolbutamide, a model CYP2C11 probe substrate, using liver microsomes, primary hepatocytes and in vivo in the rat. AB - The effects of Danshen and its active components (tanshinone I, tanshinone IIA, dihydrotanshinone and cryptotanshinone) on tolbutamide 4-hydroxylation was investigated in the rat. Danshen (0.125-2mg/ml) decreased 4-hydroxy-tolbutamide formation in vitro and in vivo. Enzyme kinetics studies showed that inhibition of tolbutamide 4-hydroxylase activity was competitive and concentration-dependent. The K(i) values of the tanshinones were: dihydrotanshinone (8.92microM), cryptotanshinone (24.5microM), tanshinone I (80.3microM) and tanshinone IIA (242.9microM). In freshly prepared primary rat hepatocytes, tanshinones inhibited tolbutamide 4-hydroxylation in a concentration-dependent manner, with EC(40) values in the order: cryptotanshinone (15.8microM), tanshinone IIA (16.2microM), dihydrotanshinone (20.1microM) and tanshinone I (48.2microM). In whole animal studies, single dose Danshen treatment (50 or 200mg/kg, i.p.) increased tolbutamide clearance (17-26.9%), decreased AUC (14.4-20.9%) and increased the Vd (7.26%). Three-day Danshen treatment (200mg/kg/day, i.p.) decreased the C(initial), increased T(1/2) and Vd but did not affect tolbutamide clearance and AUC. Tolbutamide-4-hydroxylation in vivo was decreased by Danshen after acute and after 3-day treatment, with decreases in the AUC of 4-hydroxy-tolbutamide (15 28%) over the time period studied. Despite competitive inhibition of rat CYP2C11 in vitro and in vivo, as shown by the decrease in tolbutamide 4-hydroxylation, only minor changes in tolbutamide pharmacokinetics was observed. This study illustrated that the herb-drug interaction potential should be monitored by both in vitro and in vivo biotransformation/ pharmacokinetic parameters. PMID- 19679456 TI - Anti-leukemic activity of Dillenia indica L. fruit extract and quantification of betulinic acid by HPLC. AB - The methanolic extract of Dillenia indica L. fruits showed significant anti leukemic activity in human leukemic cell lines U937, HL60 and K562. This finding led to fractionation of the methanolic extract, on the basis of polarity, in which the ethyl acetate fraction showed the highest anti-leukemic activity. A major compound, betulinic acid, was isolated from the ethyl acetate fraction by silica gel column chromatography and was identified and characterized. Betulinic acid could explain the anti-leukemic activity of the methanolic extract and the ethyl acetate fraction. Hence the quantitative estimation of betulinic acid was approached in methanolic extract and fractions using HPLC. PMID- 19679457 TI - In vivo genotoxicity evaluation of a plant based antiarthritic and anticancer therapeutic agent Boswelic acids in rodents. AB - The genotoxic potential of anti-inflammatory/anti-arthritic and anticancer plant based drug molecule Boswelic acids (BA) was studied by in vivo system. Systematic literature survey revealed that studies on the genotoxicity of BA are not available. Although reports on genotoxicity of Boswellia serrata dry extract and modified 3-O-acetyl-11-keto-beta-boswelic acid are available and these studies were conducted in in vitro systems. The earlier general toxicity study of BA has been conducted by us, revealed it to be non toxic. The genotoxicity was carried out in Wistar rats using different cytogenetic assay system-abnormalities viz. chromosomal aberrations; sperm morphology, micronuclei and comet assays. Six groups of animals, each comprised of five rats, were taken for each study. Group1 4 received BA at 125, 250, 500 and 1000 mg/kg p.o., respectively prepared as 2% gum acacia suspension, fifth group received a positive control cyclophosphamide (CP) 40 mg/kg p.o. or metronedazole (MTZ) 130 mg/kg p.o. or mercuric chloride (HgCl(2)) 0.864 mg/kg p.o. (as per the experiment requirement) whereas the sixth group kept as vehicle control. The results on the bases of the data obtained revealed that BA is quite safe as it did not show any genotoxicity at any dose level up to 1000 mg/kg. The positive controls used in different experiments showed highly significant abnormal cytogenetic changes in comparison to the control group. PMID- 19679458 TI - A simple and portable breathing circuit designed for ventilatory muscle endurance training (VMET). AB - BACKGROUND: Ventilatory muscle endurance training (VMET) involves increasing minute ventilation (V (E)) against a low flow resistance at rest to simulate the hyperpnea of exercise. Ideally, VMET must maintain normocapnia over a wide range of V (E). This can be achieved by providing a constant fresh gas flow to a sequential rebreathing circuit. The challenge to make VMET suitable for home use is to provide a source of constant fresh gas flow to the circuit without resorting to compressed gas. METHODS: Our VMET circuit was based on a commercial sequential gas delivery breathing circuit (Pulmanex Hi-Ox, Viasys Healthcare, Yorba Linda, CA USA). Airflow was provided either by a small battery-driven aquarium air pump or by the entrainment of air down a pressure gradient created by the recoil of a hanging bellows that was charged during each inhalation. In each case, fresh gas flow was adjusted to be just less than resting V (E). Eight subjects then breathed from the circuit for three 10min periods consisting of relaxed breathing, breathing at 20 and then at 40L/min. We monitored V (E), end tidal PCO2 (PetCO2) and hemoglobin O2 saturation (SpO2). RESULTS: During hyperpnea at 20 and 40L/min, PetCO2 did not differ significantly from resting levels with either method of supplying fresh gas. SpO2 remained greater than 96% during all tests. CONCLUSION: Isocapnic VMET can be reliably accomplished with a simple self-regulating, sequential rebreathing circuit without the use of compressed gas. PMID- 19679459 TI - The skinny on Fat: an enormous cadherin that regulates cell adhesion, tissue growth, and planar cell polarity. AB - Fat is an extremely large atypical cadherin involved in the regulation of cell adhesion, tissue growth, and planar cell polarity (PCP). Recent studies have begun to illuminate the mechanisms by which Fat performs these functions during development. Fat relays signals to the Hippo pathway to regulate tissue growth, and to PCP proteins to regulate tissue patterning. In this review we briefly cover the historical data demonstrating that Fat regulates tissue growth and tissue patterning, and then focus on advances in the past three years illuminating the mechanisms by which Fat controls growth and planar polarity in flies and mammals. PMID- 19679460 TI - Effectiveness of unit-based pricing of waste in the Netherlands: applying a general equilibrium model. AB - Differential and variable rates (DVR) in waste collection charging give a price incentive to households to reduce their waste and increase recycling. International empirical evidence confirms the effectiveness of DVR schemes, with limited unwanted side effects. In the Netherlands, currently some 20% of the population is charged at DVR. This is less than in several other countries. Taking into account differences between types of households and dwellings, this study analyses various scenarios for extended use of DVR in the Netherlands. The analysis shows that further penetration of DVR is a cost-effective instrument for waste reduction and more recycling. Moreover, DVR can itself be seen as a necessary condition for the successful implementation of other economic instruments, such as waste taxes. It is therefore recommended to stimulate municipalities to adopt DVR schemes in the Netherlands, accompanied by the provision of adequate facilities for waste separation by households. Before introducing DVR in 'very strongly urbanized' municipalities (i.e. the 12 largest cities in the Netherlands) a pilot experiment in one of them might be useful to test the behavioral response in this category. PMID- 19679461 TI - An improved glucose/O2 membrane-less biofuel cell through glucose oxidase purification. AB - A key objective in any bioelectrochemical systems is to improve the current densities and mass transport limitation. Most of the work is focused on increasing the specific surface of the electrodes or improving the electron transfer between enzymes and electrodes. However, nothing is said about the comparison of purified and non-purified enzyme and their effects on the biosensor efficiency. To illustrate the effect of the enzyme purity, we studied the widely used commercial Glucose Oxidase (GOx) from Aspergillus niger that we are using in our miniature membrane-less biofuel cell. Our results indicate that even if additional compounds contained in the lyophilized enzyme powder do not interfere with its intrinsic catalytic properties, they could prevent a good electron transfer between the enzyme and the electrode surface. By introducing a purified glucose oxidase into a bioelectrocatalyst immobilized on an electrode surface, we show that we can increase the interaction between the enzyme and the redox polymer, forming a better homogenous, leather like gel. At 5mM glucose concentration and under oxygen atmosphere, the current is three-fold higher when using a purified enzyme than it is when using a non-purified enzyme. Built with this novel anode, we showed that a miniature implantable membrane-less glucose O(2) biofuel cell could produce, under air, twice the power density that is usually obtained when using a non-purified GOx. PMID- 19679462 TI - Lipid-protein interactions probed by electron crystallography. AB - Electron crystallography is arguably the only electron cryomicroscopy (cryoEM) technique able to deliver an atomic-resolution structure of membrane proteins embedded in the lipid bilayer. In the electron crystallographic structures of the light driven ion pump, bacteriorhodopsin, and the water channel, aquaporin-0, sufficiently high resolution was obtained and both lipid and protein were visualized, modeled, and described in detail. An extensive network of lipid protein interactions mimicking native membranes is established and maintained in two-dimensional (2D) crystalline vesicles used for structural analysis by electron crystallography. Lipids are tightly integrated into the protein's architecture where they can affect the function, structure, quaternary assembly, and the stability of the membrane protein. PMID- 19679464 TI - Early primary breast cancer in the elderly - pattern of presentation and treatment. AB - INTRODUCTION: The incidence of primary breast cancer in elderly patients is increasing. However, little is known about their biological profile and most appropriate clinical management, as most studies have been conducted in the younger population. This study aimed to identify a profile of characteristics in elderly women with operable primary breast cancer and investigate the dynamics influencing the treatment decision-making process. METHODS: A review of 268 consecutive female patients >70 years of age, diagnosed with early operable primary breast cancer (<5 cm) over a 30-month period at the Nottingham Breast Institute, was conducted. Age, co-morbidity, cancer characteristics, treatment offered and undertaken, and reason for patient choice were recorded and analysed. RESULTS: The median age was 78 (range 70-100) years. In our study, 82% of the patients had one or more co-morbidities, with 34% of them having three or more co morbidities. The commonest pathological diagnosis (from needle core biopsies) was invasive ductal carcinoma of no special type (76%) with histological grade 2 (64%). Majority of them were oestrogen receptor (ER)-positive (84%) and had a high histochemical (H)-score (83% with H-score >200). Most of the patients (60%) underwent primary surgical management, of which 45.4% received breast-conserving surgery. Among the patients who had breast-conserving surgery, 68% of them received adjuvant radiotherapy. When offered genuine choice in treatment options, most patients chose non-operative treatment. Patients who underwent non-operative treatment were on average seven years older and had significantly more co morbidities than those who had surgery. CONCLUSION: The elderly population evidently have demographic and cancer characteristics distinct from their younger counterparts, with less patients receiving surgical management. Further work is underway to correlate this with their clinical outcomes and to examine the factors behind the treatment decision-making process. PMID- 19679463 TI - Screening of anti-glioma effects induced by sigma-1 receptor ligands: potential new use for old anti-psychiatric medicines. AB - The prognosis of glioblastoma (GBM) remains poor. Diffuse invasion of distant brain tissue by migrating cells from the primary tumour mass has already occurred at time of diagnosis. Anti-cancer effects of a selective sigma-1 agonist, 4-(N benzylpiperidin-4-yl)-4-iodobenzamide (4-IBP), in glioblastoma were shown previously, leading to the present work where the effects on glioblastoma cells of 17 agonists or antagonists of sigma-1 receptors were studied, including currently marketed drugs fluvoxamine, dextromethorphan, donepezil, memantine and haloperidol. We first showed that established GBM cell lines, primary cultures and surgical specimens express sigma-1 receptors. In vitro analyses then focused on anti-proliferation and anti-migratory effects on human glioblastoma cell lines using quantitative videomicroscopy analyses. These cell monitoring assays revealed specific impacts on the mitotic cell process. Using an aggressive glioma model orthotopically grafted into the brains of immunocompromised mice, we showed that combining donepezil and temozolomide gave additive benefit in terms of long survivors as compared to temozolomide or donepezil alone. Clinical study is planned if further rodent dose-ranging studies of donepezil with temozolomide continue to show evidence of benefit in this model. PMID- 19679465 TI - Rotary drum composting of vegetable waste and tree leaves. AB - High rate composting studies on institutional waste, i.e. vegetable wastes, tree leaves, etc., were conducted on a demonstration-scale (3.5 m(3)) rotary drum composter by evaluating changes in some physico-chemical and biological parameters. During composting, higher temperature (60-70 degrees C) at inlet zone and (50-60 degrees C) at middle zone were achieved which resulted in high degradation in the drum. As a result, all parameters including TOC, C/N ratio, CO(2) evolution and coliforms were decreased significantly within few days of composting. Within a week period, quality compost with total nitrogen (2.6%) and final total phosphorus (6 g/kg) was achieved; but relatively higher final values of fecal coliforms and CO(2) evolution, suggested further maturation. Thus, two conventional composting methods namely windrow (M1) and vermicomposting (M2) tried for maturation of primary stabilized compost. By examining these methods, it was suggested that M2 was found suitable in delivering fine grained, better quality matured compost within 20 days of maturation period. PMID- 19679466 TI - Start-up procedures and analysis of heavy metals inhibition on methanogenic activity in EGSB reactor. AB - The effectiveness of operating an industrial UASB reactor, treating wastewater from the beer industry, with flows containing heavy metals was evaluated. A pilot scale UASB reactor, already used to simulate the industrial reactor, was unsuccessfully employed. An easy start-up was obtained arranging it as an EGSB reactor. Considerations about this modification are reported. The effects of Cu(II), Ni(II) and Cr(III) ions on the anaerobic activity were analyzed by measurements of methane production rate and COD removal. The employed biomass was the sludge of the industrial UASB reactor, while a solution of ethanol and sodium acetate with COD of 3000 mg/L and a heavy metal concentration of 50 mg/L were continuously fed. Experimental results proved higher biomass sensitivity for copper and much slighter for nickel and chromium. Moreover, copper inhibition has been demonstrated to be less significant if a metal-free feed was provided to the system before copper addition. PMID- 19679467 TI - Comparative study of liquefaction process and liquefied products from bamboo using different organic solvents. AB - The effects of various solvents, including phenol, ethylene glycol (EG) and ethylene carbonate (EC), and different liquid ratios on the liquefaction of bamboo, have been studied systematically in this paper. The processes were catalyzed by hydrochloride acid at 180 degrees C in autoclaves for different reaction times. The results show that phenol is the optimum solvent for bamboo liquefaction with a yield up to 99%. The Fourier transform-infrared (FT-IR) analyses of the residues show that cellulose, hemicelluloses and lignin are almost decomposed when using phenol as solvent. The gel permeation chromatography (GPC) results of the liquid products show that the high molecular weight of bamboo decreases significantly to around 1800 g mol(-1) after liquefaction. The gas chromatography and mass spectrometry (GC-MS) analysis shows that low boiling point products of liquefied bamboo are similar regardless of the type of solvent used. PMID- 19679468 TI - Unusual intramolecular N-->O acyl group migration occurring during conjugation of (-)-DHMEQ with cysteine. AB - Previously we found that (-)-DHMEQ, a specific NF-kappaB inhibitor, covalently bound to a specific cysteine of NF-kappaB component proteins. In the course of formation of the (-)-DHMEQ and protected cysteine conjugate, we observed an unusual intramolecular N-->O acyl group migration. PMID- 19679469 TI - Identification of positron emission tomography ligands for NPY Y5 receptors in the brain. AB - A series of trans-3-oxospiro[(aza)isobenzofuran-1(3H),1'-cyclohexane]-4' carboxamide derivatives were synthesized and profiled for NPY Y5 binding affinity, brain and CSF penetrability in rats, and susceptibility to human and mouse P-glycoprotein transporters in order to develop a PET ligand. Compound 12b exhibited an acceptable profile for a PET ligand, and [(11)C]12b was successfully utilized in clinical settings as a Y5 PET ligand. PMID- 19679470 TI - Synthesis of gibberellin derivatives with anti-tumor bioactivities. AB - A series of gibberellin based molecules were designed and synthesized. Gibberellin derivatives bearing two alpha,beta-unsaturated ketone units showed strong anticancer activities in MTT assay towards a number of human cancer cell lines including HT29, A549, HepG2 and MKN28. The most potent gibberellin derivative (compound 10, IC(50)=2.9 microM against HT29) inhibited completely the topoisomerase I activity at 8 microg/mL level. PMID- 19679472 TI - Prenyloxyphenylpropanoids as a novel class of anticonvulsive agents. AB - In this study, we synthesized some natural and semi-synthetic prenyloxyphenylpropanoids (e.g., acetophenones, benzoic and cinnamic acids, chalcones, and coumarins), and we assessed their in vivo neuroprotective activity, using the mouse maximal electroshock-induced seizure model (MES test). 7-Isopentenyloxycoumarin and (2E)-3-{4-[(3-methylbut-2-enyl)oxy]phenyl}prop-2 enoic acid, administered ip at a dose of 300 mg/kg, suppressed MES-induced seizures in mice in a time- and dose-dependent manner. PMID- 19679471 TI - Chemical methods for the synthesis and modification of neoclerodane diterpenes. AB - Diterpenes are a structural class of molecules that are derived from four isoprene subunits and are widespread throughout nature. A number of neoclerodane diterpenes have been found to have biological activity but a limited number of chemical investigations have been conducted. Recently, the neoclerodane diterpene, salvinorin A (12) has been investigated due to its unique pharmacological profile. This review will discuss the chemical methods used to chemically modify and synthesize 12. PMID- 19679473 TI - WITHDRAWN: Design and synthesis of pivalic acid linked tacrine-NO donor hybrids as novel anti-Alzheimer drug candidates. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19679474 TI - Molecular modeling studies to predict the possible binding modes of endomorphin analogs in mu opioid receptor. AB - The molecular docking of a series of endomorphin analog with the mu opioid receptor was performed. The successive molecular dynamics of several proposed ligand-receptor complexes inserted into the phospholipid bilayer were carried out to optimize the complex and explore the conformational changes. Meaningful differences of their binding modes were detected and the involvement of some essential residues in ligand binding was also identified. Our proposed ligand receptor model is in good agreement with previous site-directed mutagenesis experiments. PMID- 19679475 TI - A novel class of highly potent multidrug resistance reversal agents: disubstituted adamantyl derivatives. AB - Novel disubstituted adamantyl derivatives were synthesized and evaluated in a P glycoprotein dependent multidrug resistance cancer cell line. The hit to lead optimization provided potent MDR reversal agents. Some potent adamantyl derivatives were more than 10-fold more potent than verapamil without considerable intrinsic cytotoxicity. The 3-trifluorophenyl derivative 14f did not affect the metabolism of CYP450 3A4, whereas most of MDR revertants had a weak inhibitory effect. PMID- 19679476 TI - Design and synthesis of bile acid-based amino sterols as antimicrobial agents. AB - New bile acid-based amino sterols were synthesized in good yields from C-3beta oxiranes as key intermediates. These derivatives were evaluated for their in vitro antimicrobial properties against human pathogens. These compounds showed better antibacterial activity as compared to antifungal activity. Compounds 21 and 22 showed comparable antibacterial activity to gentamicin against Staphylococcus aureus with IC50 values of 5.14 and 4.46 microg/mL. This is the first report for the synthesis of C-3beta-oxiranes on the steroids having A/B cis ring junction and these oxiranes have been used for the synthesis of amino sterols 17, 18, 21, and 22. PMID- 19679477 TI - Synthesis and evaluation of inhibitors of cytochrome P450 3A (CYP3A) for pharmacokinetic enhancement of drugs. AB - The HIV protease inhibitor ritonavir (RTV) is also a potent inhibitor of the metabolizing enzyme cytochrome P450 3A (CYP3A) and is clinically useful in HIV therapy in its ability to enhance human plasma levels of other HIV protease inhibitors (PIs). A novel series of CYP3A inhibitors was designed around the structural elements of RTV believed to be important to CYP3A inhibition, with general design features being the attachment of groups that mimic the P2-P3 segment of RTV to a soluble core. Several analogs were found to strongly enhance plasma levels of lopinavir (LPV), including 8, which compares favorably with RTV in the same model. Interestingly, an inverse correlation between in vitro inhibition of CYP3A and elevation of LPV was observed. The compounds described in this study may be useful for enhancing the pharmacokinetics of drugs that are metabolized by CYP3A. PMID- 19679478 TI - An unusual presentation of muscle-eye-brain disease: severe eye abnormalities with mild muscle and brain involvement. AB - Muscle-eye-brain disease (MEB) is characterised by congenital muscular dystrophy, structural brain malformations and eye abnormalities. We report a MEB case whose presenting sign was congenital blindness. She was investigated primarily for eye abnormalities at onset. She had bilateral retinal detachment and microphthalmia. Mild axial hypotonia and motor retardation were attributed to cerebral disorder in another center. Muscle biopsy showed mild myopathic changes and significant alpha-dystroglycan deficiency. Analysis of the POMGnT1 showed a novel homozygous mutation 1814G>C, causing p.Arg605Pro change. This case expands the clinical spectrum of MEB with unusually severe eye abnormalities compared to mild skeletal muscle and brain involvement. PMID- 19679479 TI - Review of quantitative measurements of upper limb movements in hemiplegic cerebral palsy. AB - This review provides an overview of results found in literature on objective measurements of upper limb movements in children with hemiplegic cerebral palsy (HCP). Seventeen articles were selected following a systematic search. Analysed tasks varied from simple reaching and gross motor functions to complex, fine motor tasks. Spatiotemporal characteristics have been extensively studied and longer movement durations, slower movement speed and reduced trajectory straightness at the affected upper limb, compared to the non-affected side or healthy children, were most frequently reported. Joint kinematics has been far less studied. The limited data confirm the clinical impression of children with HCP using less elbow extension and supination to reach for an object, which is compensated by increased trunk flexion. Increased trunk involvement was also reported during gross motor functions. Although three-dimensional (3D) movement analysis seems promising to provide additional insights in the pathological upper limb movements observed in HCP, future standardisation of the entire protocol is crucial. No consensus exists on the procedures for data collection, processing, analysing and reporting of results, or what upper limb tasks should be assessed. The International Society of Biomechanics recently proposed recommendations on the definition of upper limb joint coordinate systems and rotation sequences. These guidelines were not yet applied in these studies. Although the diverse methodological approaches used in the studies complicate the comparison of published results, some general conclusions could be drawn. A further standardisation of the protocol for 3D upper limb movement analysis will provide the foundation for comparable and repeatable results and eventually facilitate the selection and planning of treatment interventions. PMID- 19679480 TI - What did Darwin say about microbes, and how did microbiology respond? AB - Although it is commonly assumed that Darwin had nothing to say about microbes, he did in fact say quite a lot. He included microbes in his Beagle studies of the geographical distribution of organisms and used microscopic organisms as explicit exemplars of how adaptation did not imply increasing complexity. Darwin often discussed microorganismal classification, origins and experimentation in his correspondence. But despite his interests in microbial phenomena, Darwin's impact on microbiological thinking of the late nineteenth century was negligible. This limited response may be connected to today's assumptions about Darwin's neglect of microbes. PMID- 19679481 TI - Fluorinated indazoles as novel selective inhibitors of nitric oxide synthase (NOS): synthesis and biological evaluation. AB - In order to find new compounds with neuroprotective activity and NOS-I/NOS-II selectivity, we have designed, synthesized, and characterized 14 new NOS inhibitors with an indazole structure. The first group corresponds to 4,5,6,7 tetrahydroindazoles (4-8), the second to the N-methyl derivatives (9-12) of 7 nitro-1H-indazole (1) and 3-bromo-7-nitro-1H-indazole (2), and the latter to 4,5,6,7-tetrafluoroindazoles (13-17). Compound 13 (4,5,6,7-tetrafluoro-3-methyl 1H-indazole) inhibited NOS-I by 63% and NOS-II by 83%. Interestingly, compound 16 (4,5,6,7-tetrafluoro-3-perfluorophenyl-1H-indazole) inhibited NOS-II activity by 80%, but it did not affect to NOS-I activity. Structural comparison between these new indazoles further supports the importance of the aromatic indazole skeleton for NOS inhibition and indicate that bulky groups or N-methylation of 1 and 2 diminish their effect on NOS activity. The fluorination of the aromatic ring increased the inhibitory potency and NOS-II selectivity, suggesting that this is a promising strategy for NOS selective inhibitors. PMID- 19679482 TI - N-[2-chloro-9H-purin-6-yl]-N-cyclopropylglycylamino acids and derivatives: synthesis, evaluation as a class of novel analgesics, and 3D QSAR analysis. AB - Via a five-step-reaction procedure for the preparation of 19 known N-[2-chloro-9 (tetrahydropyran-2-yl)-9H-purin-6-yl]-N-cyclopropylglycylamino acid benzylesters (6a-s) and successive removal of 9-(tetrahydropyran-2-yl) and benzylester groups 19 novel N-[2-chloro-9H-purin-6-yl]-N-cyclopropylglycylamino acid benzylesters (7a-s) and 19 novel N-[2-chloro-9H-purin-6-yl]-N-cyclopropylglycylamino acids (8a s) were provided. On tail-flick mouse model the in vivo analgesic activities of these 38 novel compounds were measured and most of them were defined as good analgesics. Based on Molecular Field Analysis of the pain threshold variations of the mice receiving 48 compounds in terms of the descriptors proton and methyl an equation was established. The data points (n), correlation coefficient (r), and square correlation coefficient (r(2)) of this equation were 48, 0.923, and 0.852, respectively. Using this equation pain threshold variations of 9 compounds were predicted and the errors ranged from 1.71 to 8.92. PMID- 19679483 TI - Synthesis and evaluation of anticancer activity of 2-arylamino-6-trifluoromethyl 3-(hydrazonocarbonyl)pyridines. AB - The synthesis and anticancer activity of 2-arylamino-6-trifluoromethyl-3 (hydrazonocarbonyl)pyridines is described. The new trifluoromethylpyridine derivatives were evaluated for their anticancer activity toward human cancer cell lines by the National Cancer Institute (NCI). Most of them had excellent growth inhibition activity, having GI(50) values in the low micromolar to nanomolar concentration range. The most potent 2,6-dichlorobenzaldehydehydrazone 29 inhibited the growth of all tested cancer cell lines with nanomolar potency, and did not show animal toxicity. Hydrazone 29 has been selected by the Biological Evaluation Committee of NCI for testing in vivo Hollow Fiber Assay. PMID- 19679484 TI - Preparation of conophylline affinity nano-beads and identification of a target protein. AB - Conophylline, a vinca alkaloid extracted from the tropical plant Ervatamia microphylla, has been shown to induce the differentiation of insulin-producing beta-cells in cultured cells and in animals. However, its mechanism of action and the molecular target have remained unclear. Therefore, we prepared a fishing probe with conophylline to identify the target protein by using latex nano-beads, which are newly innovated tools for affinity-purification. With these conophylline-linked nano-beads, we found that conophylline directly interacted with ARL6IP. ARL6IP may thus be involved in the mechanism of cellular differentiation of beta-cells, and this probe should be useful to find other target proteins. PMID- 19679485 TI - Structural and biological evaluation of some loloatin C analogues. AB - Loloatin C is a cyclic cationic antimicrobial peptide which is active against gram positive as well as certain gram negative bacteria. Unfortunately, it is equally potent against human erythrocytes. To probe the structure-activity relationship of this promising antibiotic peptide, amino acid substitution and/or incorporation of a constraint sugar amino acid dipeptide isoster has been applied. Six new derivatives have been synthesized using SPPS and their solution structure investigated using NMR studies. Finally, the antimicrobial and the hemolytic activities have been determined. PMID- 19679486 TI - [(18)F]-labeled 2-methoxyphenylpiperazine derivative as a potential brain positron emission tomography imaging agent. AB - This study reports the synthesis and characterization of N-(3-(4-(2 methoxyphenyl)piperazin-1-yl)propyl-4-[(18)F]fluorobenzamide ([(18)F]MPP3F). The total reaction time for [(18)F]MPP3F, including final high-performance liquid chromatography purification, was about 3h. Typical decay-corrected radiochemical yield was 18.4+/-3.1%. The radiochemical purity was >98%. Biodistribution in mice showed that [(18)F]MPP3F is a potential brain imaging agent for positron emission tomography. The brain uptake of [(18)F]MPP3F was 6.59+/-0.77% Injected Dose/g at 2 min post-injection time. A brain-to-blood ratio of 3.67 was reached at 15 min after injection. PMID- 19679487 TI - Crystal rounding and the efficiency transfer method in gamma-ray spectrometry. AB - The effect of failing to properly model the rounding of the crystal edges in the application of the efficiency transfer method to the measurement of environmental samples is investigated for coaxial and thick planar (BeGe) HPGe detectors. It is found that for lower energies proper modelling of the crystal rounding is necessary to achieve the accuracy of the computed efficiency usually required in this type of measurements. PMID- 19679489 TI - Synthesis, photophysical properties and sugar-dependent in vitro photocytotoxicity of pyrrolidine-fused chlorins bearing S-glycosides. AB - Eight S-glycosylated 5,10,15,20-tetrakis(tetrafluorophenyl)porphyrins (1a', 1b', 1a and 1b (a: S-glucosylated, b: S-galactosylated)) and their 1,3-dipolar cycloadducts, i.e. chlorins 2a', 2b', 2a and 2b were prepared by nucleophilic substitution of the pentafluorophenyl groups with S-glycoside. These photosensitizers were characterized by (1)H, (13)C and (19)F NMR spectroscopies and elemental analysis. The photocytotoxicity of the S-glycosylated photosensitizers and the parent porphyrin (1) and chlorin (2) was examined in HeLa cells. Photosensitizers 1, 2, 1a', 1b', 2a' and 2b' showed no significant photocytotoxicity at the concentration of 0.5microM, while the deprotected photosensitizers 1a, 1b, 2a and 2b were photocytotoxic. The strong inhibition by sodium azide of the photocytotoxicity of these photosensitizers suggested that (1)O(2) is the main mediator. The S-glucosylated photosensitizers 1a and 2a showed higher photocytotoxicity than S-galactosylated 1b and 2b, respectively. The cellular uptake of 1a and 2a increased up to 24h, while that of 1b and 2b was saturated by 12h. PMID- 19679488 TI - Trends of bacterial colonisation and the risk of postoperative pneumonia in lung cancer patients with chronic obstructive pulmonary disease. AB - BACKGROUND: Lung cancer patients with chronic obstructive pulmonary disease (COPD) have a high risk of developing postoperative pneumonia (POP). This study aims to investigate the impact of COPD on POP and the trends for perioperative bronchial colonisation by micro-organisms. METHODS: A retrospective chart review was made for 626 patients who underwent lung cancer surgeries at the Chiba University Hospital between 1996 and 2005. The patients were categorised as non COPD (n=475) and COPD (forced expiratory volume in 1s/forced vital capacity (FEV1/FVC) <70%; n=151). All the patients had sputum and bronchial bacterial cultures examined for potentially pathogenic micro-organisms (PPMs). Risk factors for POP and mortality were analysed. RESULTS: Patients with COPD had a significantly higher incidence of POP (23/151, 15.2%) than those without COPD (17/475, 3.6%) (p<0.0001). Preoperative bronchial bacterial examinations showed that 50 of 475 patients without COPD (10.5%) had positive cultures, while the results for 30 of 151 patients with COPD (19.9%) were positive (p=0.0111). Only 31 of 548 patients (5.7%) who did not show any preoperative PPMs had POP, while nine of 78 patients (11.5%) who presented preoperative PPMs had POP (p=0.0469). The PPMs that emerged postoperatively were primarily Staphylococcus aureus (and Gram-negative bacilli (94.4% of PPMs), while they were seen less frequently preoperatively (46.5% of PPMs). Multivariate analysis demonstrated that advanced age and FEV1/FVC were independent risk factors for POP. Patients with POP had significantly worse long-term survivals than those without POP (p=0.0004). CONCLUSION: COPD was a risk factor for POP. Staphylococcus aureus and Gram negative bacilli should be targets for postoperative prophylactic antibiotic selection. Patients with POP had poor long-term survivals. PMID- 19679490 TI - Lung recruitment--a guide for clinicians. AB - Recruitment manoeuvres play an important role in minimising ventilator associated lung injury (VALI) particularly when lung protective ventilation strategies are employed and as such clinicians should consider their application. This paper provides evidence-based recommendations for clinical practice with regard to alveolar recruitment. It includes recommendations for timing of recruitment, strategies of recruitment and methods of measuring the efficacy of recruitment manoeuvres and contributes to knowledge about the risks associated with recruitment manoeuvres. There are a range of methods for recruiting alveoli, most notably by manipulating positive end expiratory pressure (PEEP) and peak inspiratory pressure (PIP) with consensus as to the most effective not yet determined. A number of studies have demonstrated that improvement in oxygenation is rarely sustained following a recruitment manoeuvre and it is questionable whether improved oxygenation should be the clinician's goal. Transient haemodynamic compromise has been noted in a number of studies with a few studies reporting persistent, harmful sequelae to recruitment manoeuvres. No studies have been located that assess the impact of recruitment manoeuvres on length of ventilation, length of stay, morbidity or mortality. Recruitment manoeuvres restore end expiratory lung volume by overcoming threshold opening pressures and are most effective when applied after circuit disconnection and airway suction. Whether this ultimately improves outcomes in adult or paediatric populations is unknown. PMID- 19679491 TI - Acid hydrolysis of proteins in matrix assisted laser desorption ionization matrices. AB - Sample preparation is crucial to the success of experiments in biological mass spectrometry. In proteomics, digestion of the proteins into peptides is a key step for "bottom-up" approaches. Often, the use of enzymes requires physiological conditions, producing peptides that must be extracted or further purified before mass analysis. Chemical cleavage reagents offer more flexibility and can be more compatible with downstream mass analysis. Expanding on prior work using acid hydrolysis, proteolysis with matrix-assisted laser desorption ionization (MALDI) matrices is presented. This sample preparation can be performed rapidly with a minimum of reagents and sample handling, but it must first be evaluated in terms of digestion efficiency, missed cleavages, and side reactions before implementation for in-gel digestion and in-solution digestion using minimal volumes of protein. Time courses of acid hydrolysis are shown for protein standards, illustrating the sensitivity of this type of sample preparation, minimization of side reactions, and performance for proteins in mixtures. To illustrate the potential for sensitive detection of a specific protein, MALDI matrix hydrolysis is used to digest a protein immunoprecipitated from cell lysate. PMID- 19679492 TI - Exposure to high-risk genital human papillomavirus and its association with risky sexual practices and laboratory-confirmed chlamydia among African-American women. AB - BACKGROUND: Genital human papillomavirus (HPV) is the most common sexually transmitted infection (STI) in the United States and African-American women have the highest prevalence of high-risk HPV. This study examined exposure to high risk HPV in African-American women and its relation to risky sexual practices and laboratory-confirmed chlamydia. METHODS: A sample of 665 African-American women between 18 and 29 years old, recruited from October 2002 to March 2006 in Atlanta, Georgia, completed an Audio Computer-Assisted Survey Interview assessing sociodemographics, health practices, and risky sexual practices. Participants also provided vaginal swab specimens assayed for STIs and high-risk HPV. RESULTS: The overall prevalence of high-risk HPV was 38.9%. Among women 18 to 24 years old, it was 42.4%; it was 31% among women 25 to 29 years old. Age-stratified logistic regression analyses indicated that women between the ages of 18 and 29 and 18 and 24 who had multiple male sexual partners did not use a condom during their last casual sexual encounter and tested positive for chlamydia were significantly more likely to test positive for high-risk HPV. Women 18 to 24 years old who reported having a casual or risky sexual partner were significantly more likely to test positive for high-risk HPV. No significant correlates were identified among women 25 to 29 years old. CONCLUSIONS: Programs should aim to educate, decrease risky sexual practices, and increase screening and treatment for STIs among women with high-risk HPV infections. HPV vaccination recommendations for young adult African-American women warrant special consideration. PMID- 19679493 TI - Time-related changes of motor unit properties in the rat medial gastrocnemius muscle after spinal cord injury. I. Effects of total spinal cord transection. AB - The contractile properties of motor units (MUs) were electrophysiologically investigated in the medial gastrocnemius (MG) muscle in 17 Wistar three-month-old female rats: 14, 30, 90 and 180 days after the total transection of the thoracic spinal cord and compared to those in intact (control) rats. A sag phenomenon, regularly observed in unfused tetani of fast units in intact animals at 40 Hz stimulation, almost completely disappeared in spinal rats. Therefore, the MUs of intact and spinal rats were classified as fast or slow types basing on 20 Hz tetanus index, the value of which was lower or equal 2.0 for fast and higher than 2.0 for slow MUs. The MUs composition of MG muscle changed with time after the spinal cord transection: an increasing proportion of fast fatigable (FF) units starting one month after injury and a disappearance of slow (S) units within the three months were observed. In all MUs investigated the twitch contraction and half-relaxation time were significantly prolonged after injury (p<0.01, Mann Whitney U-test). Moreover, a decrease of the fatigue index for fast resistant (FR) and slow MUs was observed in subsequent groups of spinal rats. No significant changes were found between twitch forces in all MU types of spinal animals (p>0.05). However, due to a decrease of the maximal tetanic force, a significant rise of the twitch-to-tetanus ratio of all MUs in spinal rats was detected (p<0.01). The considerable reduction of ability to potentiate the force was noticed for fast, especially FF type MUs. In conclusion, the spinal cord transection leads to changes in the proportion of the three MU types in rat MG muscle. The majority of changes in MUs' contractile properties were developed progressively with time after the spinal cord injury. However, the most intensive alterations of twitch-time parameters were observed in rats one month after the transection. PMID- 19679494 TI - The effect of provocative tests on electrodiagnosis criteria in clinical carpal tunnel syndrome. AB - INTRODUCTION AND OBJECTIVE: Nerve conduction study is the most sensitive test for diagnosis of carpal tunnel syndrome (CTS). This test is normal in some patients with mild CTS. Median nerve conduction study evaluation after a provocative test (e.g. wrist flexion) may be helpful for diagnosis of mild CTS. This study aimed to determine the effect of wrist flexion on median nerve conduction in patients suspected to CTS and in healthy subjects. MATERIALS AND METHODS: In this case control study, 20 patients (20 hands) with clinical signs of CTS and normal routine electrodiagnosis test results and 20 healthy subjects were investigated. Measured parameters included: median nerve distal sensory latency (DSL), nerve conduction velocity (NCV) across wrist, compound nerve action potential (CNAP), distal motor latency (DML) and compound muscle action potential amplitude (CAMPAMP). The above noted parameters were measured before and after 5min of full wrist flexion. Data were analyzed using paired T-test. RESULTS: Distal sensory latency increment and NCV decrimental after 5 min of wrist flexion in the patients group were statistically significant (p<0.01). The same parameters did not show significant incremental or detrimental changes in the control group. CONCLUSION: Median nerve DSL and NCV measurement after 5 min of wrist flexion may be helpful in determining more sensitive parameters in the electrodiagnosis of CTS. PMID- 19679495 TI - Time-related changes of motor unit properties in the rat medial gastrocnemius muscle after the spinal cord injury. II. Effects of a spinal cord hemisection. AB - The contractile properties of motor units (MUs) were investigated in the medial gastrocnemius (MG) muscle in rats after the spinal cord hemisection at a low thoracic level. Hemisected animals were divided into 4 groups: 14, 30, 90 and 180 days after injury. Intact rats formed a control group. The mass of the MG muscle did not change significantly after spinal cord hemisection, hind limb locomotor pattern was almost unchanged starting from two weeks after injury, but contractile properties of MUs were however altered. Contraction time (CT) and half-relaxation time (HRT) of MUs were prolonged in all investigated groups of hemisected rats. The twitch-to-tetanus ratio (Tw/Tet) of fast MUs after the spinal cord hemisection increased. For slow MUs Tw/Tet values did not change in the early stage after the injury, but significantly decreased in rats 90 and 180 days after hemisection. As a result of hemisection the fatigue resistance especially of slow and fast resistant MU types was reduced, as well as fatigue index (Fat I) calculated for the whole examined population of MUs decreased progressively with the time. After spinal cord hemisection a reduced number of fast MUs presented the sag at frequencies 30 and 40 Hz, however more of them revealed sag in 20 Hz tetanus in comparison to control group. Due to considerable changes in twitch contraction time and disappearance of sag effect in unfused tetani of some MUs in hemisected animals, the classification of MUs in all groups of rats was based on the 20 Hz tetanus index (20 Hz Tet I) but not on the standard criteria usually applied for MUs classification. MU type differentiations demonstrated some clear changes in MG muscle composition in hemisected animals consisting of an increase in the proportion of slow MUs (likely due to an increased participation of the studied muscle in tonic antigravity activity) together with an increase in the percentage of fast fatigable MUs. PMID- 19679497 TI - Extracranial blood flow distribution during carotid surgery. AB - OBJECTIVE: The collateral function of the external carotid artery (ECA) for cerebral perfusion in cases of atherosclerotic occlusive disease of the internal carotid artery (ICA) is difficult to assess; for this reason, blood flow measurements were taken during carotid endarterectomy (CEA). METHODS: Blood flow was measured before and after CEA using a transit-time flow meter at the carotid artery in 1000 patients who underwent CEA for high-degree (>70%) ICA stenosis. The data were collected prospectively and analysed retrospectively. RESULTS: Median ICA blood flow increased significantly, up 46% from 160 ml min(-1) (IQR: 100-234 ml min(-1)) before CEA to 240 ml min(-1) (IQR: 187-309 ml min(1)) after CEA (P<0.001). Median ECA blood flow dropped by 4%, from 152 ml min(-1) (IQR: 108 220 ml min(-1)) to 150 ml min(-1) (IQR: 103-200 ml min(-1)) (P=0.001). Relative ICA blood flow volumes related to common carotid artery (CCA) flow increased from 58% before CEA to 73% after CEA, whereas relative ECA flow decreased from 54% to 44%. CONCLUSIONS: Increased blood flow in the ICA after CEA is accompanied by decreased ECA flow whereupon the absolute amount of this redistribution is relatively limited. A more profound evaluation of these haemodynamic conditions demands further study. PMID- 19679496 TI - Microscopic mild focal cortical dysplasia in temporal lobe dual pathology: an electrocorticography study. AB - BACKGROUND: Associations between electrophysiological and histological findings might provide an insight into the epileptogenicity of mild focal cortical dysplasia (FCD) in patients with temporal lobe epilepsy (TLE) and a dual pathology. SUBJECTS AND METHODS: A total of 22 patients with pharmacoresistant TLE were included in the study, 16 of them with histologically confirmed hippocampal sclerosis (HS) associated with neocortical temporal mild Palmini Type I FCD subtypes and 6 with HS. Intraoperative electrocorticography (ECoG) recordings were analysed for epileptiform discharge frequency and morphology. Associations between histological, and electrocorticography pattern findings in these patients were analysed. Electroclinical outcomes in these patients were also evaluated. RESULTS: Neocortical areas with mild Palmini Type-I FCD showed a significantly higher spike frequency (SF) recorded in the inferior temporal gyrus than those neocortical areas in patients with HS. There was a tendency to higher spike frequency and lower amplitude in neocortical areas with histopathologic subtype IB FCD in relation with IA during intraoperative ECoG. Post-SF excision and amplitude were significantly lower during neocortical post-excision intraoperative ECoG than during neocortical pre-excision recording. There was no difference found in the clinical outcome between patients with and without FCD. CONCLUSIONS: Intraoperative electrocorticographic interictal spike frequency recorded in the neocortical inferior temporal gyrus may help to characterize the histopathologic subtypes of mild Palmini Type-I FCD in patients with temporal lobe epilepsy (TLE) and a dual pathology. Our data support the epileptogenicity of neocortical mild FCD in TLE and assessments of ECoG patterns are relevant to determine the extent of the resection in these patients which can influence the electroclinical outcome. PMID- 19679498 TI - Depressive symptoms and birth outcomes among pregnant teenagers. AB - STUDY OBJECTIVE: Few studies have examined the effects of maternal depressive symptoms among adolescent women. The purpose of this study was to investigate the impact of depressive symptoms on birth outcomes of infants born to adolescent mothers. DESIGN: The medical records of pregnant adolescent patients were examined. Information about maternal depressive symptoms and birth outcomes was collected. SETTING: Data were collected at Washington Hospital Center, a nonprofit, community-based hospital that serves residents throughout the Washington, DC area. PARTICIPANTS: Participants were 294 African-American and Latina adolescent mothers. Mean age was 16.2 years (standard deviation [SD] 1.4). Based on self-reports of depressive symptoms, adolescents were categorized by the following: no reported symptoms, depressive symptoms without SI/SA (suicidal ideation or attempt), and depressive symptoms with SI/SA. MAIN OUTCOME MEASURES: Infant birth weight and gestational age at delivery. RESULTS: Over one-quarter of pregnant adolescents in this study reported symptoms of depression. Adolescents reporting depressive symptoms with SI/SA delivered babies that weighed 239.5 grams (98.3% confidence interval [CI] 3.9 to 475.1) less than babies born to mothers reporting depressive symptoms without SI/SA. There was no association between reported symptoms and gestational age. CONCLUSIONS: Results suggest that compared to nonpregnant teens and adults, pregnant teens may have an increased risk for depression. Additionally, pregnant adolescents with suicidal ideation are at greater risk for delivering infants of lower birth weight compared with teens reporting depressive symptoms without SI/SA and teens reporting no symptoms. This study supports the need for early screening and treatment of depression for young pregnant women. PMID- 19679499 TI - Age at menarche and menstrual irregularities of adolescents with type 1 diabetes. AB - STUDY OBJECTIVE: The aim of the study was to investigate the age at menarche and menstrual irregularities in adolescents with type 1 diabetes. DESIGN: This was an aged-matched, controlled study. SETTING: The study was done in the diabetes center of a children's hospital in Athens and in high schools in major regions of central Greece. PARTICIPANTS: The study involved two groups: a group of 100 female adolescents, 12 to 18 years of age, with type 1 diabetes; and a control group of 225 healthy adolescents, matched for age. MAIN OUTCOME MEASURES: The relationship between age at menarche and menstrual irregularities was studied. RESULTS: The mean age at menarche differed significantly in the two groups (P=0.000). In particular, the mean age at menarche in the study group was 12.2 years (+/- 1.4) and in the control group it was 11.7 years old (+/- 1.2). The mean age of menarche was older in the subgroup of adolescents who had received a diagnosis of diabetes before the age of 10 than it was in those diagnosed after the age of 10. CONCLUSIONS: Menarche was observed to be delayed in adolescents with type 1 diabetes who had been diagnosed before the age of 10 in comparison with those diagnosed after the age of 10. Menstrual irregularities were found to be more common in adolescents with type 1 diabetes than in healthy ones. PMID- 19679500 TI - Impact of chronic inflammation on the pharmacokinetic-pharmacodynamic relationship of naproxen. AB - OBJECTIVES: The use of biomarkers for predicting the clinical doses of analgesic drugs relies on the understanding of the relationship between drug exposure and response under disease conditions. In this study, we demonstrate the relevance of such a relationship for COX-inhibitors by modelling the effect of naproxen on prostaglandin E2 (PGE(2)) and thromboxane B2 (TXB(2)) in a chronic inflammation model in rats. METHODS: Rats were treated with Freund's complete adjuvant (FCA) by intraplantar injection. On post-inoculation days (PID) 7-21, animals received single or chronic (qd until day 21) doses of naproxen (10mg/kg). Blood samples were collected at various intervals after dosing to characterise naproxen pharmacokinetics and its effects on PGE(2) and TXB(2) production. PK-PD modelling was performed using nonlinear mixed effects in NONMEM. RESULTS: The inhibition of PGE(2) and TXB(2) could be described by a sigmoid E(max) model. A decrease in the potency estimates of both biomarkers was observed under chronic inflammation, as compared to healthy animals. IC(50) values for PGE(2) inhibition showed a shift from 2840+/-510 to 4000+/-677ng/ml(mean+/-SD), whilst IC(50) values for TXB(2) inhibition increased from 1180+/-323 to 3360+/-453ng/ml in healthy and FCA inoculated animals, respectively. CONCLUSIONS: Our results show that chronic inflammation causes a significant change in the potency estimates for COX inhibition. These findings illustrate the implications of pathophysiological processes on pharmacodynamics and consequently on the required exposure levels for achieving response during chronic treatment. PMID- 19679501 TI - Comparison of plasma clearance of exogenous creatinine, exo-iohexol, and endo iohexol over a range of glomerular filtration rates expected in cats. AB - The study investigated plasma clearance of exogenous creatinine (PECCT), exo iohexol (PexICT) and endo-iohexol (PenICT) in six healthy cats, four cats with chronic kidney disease (CKD) and six hyperthyroid (HT) cats to assess potential differences in glomerular filtration rate (GFR) measurement over a wide range of GFR values. The PECCT, PexICT and PenICT were performed in a combined protocol. There was a significant difference between PexICT and PenICT and PECCT in healthy cats. Differences between clearance techniques are suggested to be correlated to range in GFRs and should be taken into account when GFR is measured. PMID- 19679502 TI - Rapid and efficient synthesis of fused heterocyclic pyrimidines under ultrasonic irradiation. AB - Some fused heterocyclic pyrimidines have been synthesized in high yields using ultrasound irradiation in a one-pot, three-component and efficient process by condensation reaction of barbituric acids, aldehydes and a series of enamines in water. Prominent among the advantages of this new method are operational simplicity, good yields in short reaction times and easy work-up procedures employed. PMID- 19679503 TI - A descriptive study of the force and displacement profiles of the toggle-recoil spinal manipulative procedure (adjustment) as performed by chiropractors. AB - The aim of this study was to determine the variability of the thrust parameters produced by practitioners performing a high velocity spinal manipulative therapy technique (toggle-recoil) normally applied to the neck. Fourteen participants performed three thrust trials, separated by >30minutes, on a patient simulation device. Force and displacement generated during the thrusts were simultaneously recorded and analysed off line. Peak thrust force ranged from 18.2 to 246N with a mean of 111.2N (SD 48.8). Time to peak thrust force ranged from 20 to 100ms, mean 67.5 ms (SD 13.1). Peak thrust displacement ranged from 6.1 to 28.9mm, mean 24.1mm (SD 4.9) and time to peak thrust displacement ranged from 22.5 to 105ms, mean 59.4ms (SD 13.8). This study demonstrates that the force and displacement induced by any individual practitioner on a simulator can vary by up to 50% during a toggle-recoil thrust. Furthermore, different practitioners may vary in their force by as much 100% and in displacement by 50% when the toggle-recoil spinal manipulative procedure is performed. PMID- 19679504 TI - Clinical indicators of 'nociceptive', 'peripheral neuropathic' and 'central' mechanisms of musculoskeletal pain. A Delphi survey of expert clinicians. AB - The clinical criteria by which clinicians determine mechanisms-based classifications of pain are not known. The aim of this study was to generate expert consensus-derived lists of clinical criteria suggestive of a clinical dominance of 'nociceptive', 'peripheral neuropathic' and 'central' mechanisms of musculoskeletal pain. A web-based 3 round Delphi survey method was employed as an expert consensus building technique. One hundred and three clinical experts (31 Pain consultants, 72 musculoskeletal physiotherapists) were surveyed. Participants were asked to suggest clinical indicators of three separate categories of pain mechanisms (Round 1), then rate (Round 2) and re-rate their level of agreement/disagreement (Round 3) with those clinical indicators. Consensus was defined by a >or=80% level of agreement. Sixty-two (Response rate, 60%), 60 (58%) and 59 (57%) respondents replied to Rounds 1, 2 and 3 respectively. Twelve 'nociceptive', 14 'peripheral neuropathic' and 17 'central' clinical indicators reached consensus. These expert consensus-derived lists of clinical indicators of 'nociceptive', 'peripheral neuropathic' and 'central' mechanisms of musculoskeletal pain provide some indication of the criteria upon which clinicians may base such mechanistic classifications. Further empirical testing is required in order to evaluate the discriminative validity of these clinical criteria in particular and of mechanisms-based approaches in general. PMID- 19679506 TI - Prone hip extension with lower abdominal hollowing improves the relative timing of gluteus maximus activation in relation to biceps femoris. AB - Hamstring injuries are common. The hamstrings and gluteus maximus (GM) work as synergists during hip extension. When contraction of GM is delayed relative to Biceps Femoris (BF) the hamstrings may be predisposed to injury due to increased demand. This study investigated whether specific training affected neuromuscular control of BF and GM during Prone Hip Extension (PHE). Twenty healthy volunteers were randomly allocated to the intervention or the control group. Mean onset times for BF and GM during PHE were recorded via surface electromyography (sEMG). The intervention group performed a 10min exercise with focus on proximal to distal muscle activation involving abdominal hollowing and active GM contraction prior to PHE. The control group undertook an exercise which included only PHE. Post exercise descriptive analysis indicated that the intervention exercise reduced the delay of GM firing relative to BF, however, when baseline differences between the two groups are taken into account as a covariant, the difference attributable to the intervention is not statistically significant (P=0.166). The results suggest that a short duration exercise intervention has the potential to alter the timing of activation of GM relative to BF during PHE but this needs validation through future research. PMID- 19679505 TI - Exercise-induced mechanical hypoalgesia in musculotendinous tissues of the lateral elbow. AB - The aim of this study was to investigate mechanical sensitivity responses at the lateral elbow to repeated weekly bouts of low load exercise in healthy subjects. Thirteen young men (n=6) and women participated in 4 weeks of exercise. Arms were randomly allocated to an eccentric-only exercise protocol (ECC: 5 sets of 20 contractions) or to a concentric-eccentric protocol (CON-ECC: 5 sets of 10 eccentric/10 concentric contractions) performed at 30% maximal wrist extension force. Arms were exercised consecutively within each supervised weekly session. Quantitative measures of pressure pain threshold (PPT) recorded at three sites and maximal force for grip and wrist extension were assessed at baseline, and immediately pre/post exercise at each session. Muscle endurance during 100 maximal grip contractions force was assessed at baseline and one week following the final exercise session. Results showed that regardless of protocol, repeated low load exercise resulted in a time-dependent increase in PPT at all sites post exercise Weeks 3 and 4 and persisting at follow up Week 5 (P<0.02). No significant difference between protocols was evident for any measure. Muscle force and endurance were not significantly augmented compared with baseline. In conclusion mechanical hypoalgesia is induced by repeated low load exercises regardless of exercise mode, and this may prove beneficial if replicated clinically. PMID- 19679507 TI - Statistical models of sets of curves and surfaces based on currents. AB - Computing, visualizing and interpreting statistics on shapes like curves or surfaces is a real challenge with many applications ranging from medical image analysis to computer graphics. Modeling such geometrical primitives with currents avoids to base the comparison between primitives either on a selection of geometrical measures (like length, area or curvature) or on the assumption of point-correspondence. This framework has been used relevantly to register brain surfaces or to measure geometrical invariants. However, while the state-of-the art methods efficiently perform pairwise registrations, new numerical schemes are required to process groupwise statistics due to an increasing complexity when the size of the database is growing. In this paper, we propose a Matching Pursuit Algorithm for currents, which allows us to approximate, at any desired accuracy, the mean and modes of a population of geometrical primitives modeled as currents. This leads to a sparse representation of the currents, which offers a way to visualize, and hence to interpret, such statistics. More importantly, this tool allows us to build atlases from a population of currents, based on a rigorous statistical model. In this model, data are seen as deformations of an unknown template perturbed by random currents. A Maximum A Posteriori approach is used to estimate consistently the template, the deformations of this template to each data and the residual perturbations. Statistics on both the deformations and the residual currents provide a complete description of the geometrical variability of the structures. Eventually, this framework is generic and can be applied to a large range of anatomical data. We show the relevance of our approach by describing the variability of population of sulcal lines, surfaces of internal structures of the brain and white matter fiber bundles. Complementary experiments on simulated data show the potential of the method to give anatomical characterization of pathologies in the context of supervised learning. PMID- 19679508 TI - Synthesis, spectral characterization, catalytic and antibacterial studies of new Ru(III) Schiff base complexes containing chloride/bromide and triphenylphosphine/arsine as co-ligands. AB - A new Ru(III) Schiff base complexes of the type [RuX(EPh(3))L] (X = Cl/Br; E = P/As; L = dianion of the Schiff bases were derived by the condensation of 1,4 diformylbenzene with o-aminobenzoic acid/o-aminophenol/o-aminothiophenol in the 1:2 stoichiometric ratio) have been synthesized from the reactions of [RuX(3)(EPh(3))(3)] with appropriate Schiff base ligands in benzene in the 2:1 stoichiometric ratio. The new complexes have been characterized by analytical, spectral (IR, electronic, (1)H, (13)C NMR and ESR), magnetic moment and electrochemical studies. An octahedral structure has been tentatively proposed for all these new complexes. All the new complexes have been found to be better catalyst for the oxidation of alcohols using molecular oxygen as co-oxidant at ambient temperature and aryl-aryl coupling reactions. These complexes were also subjected to antibacterial activity studies against Escherichia coli, Aeromonas hydrophilla and Salmonella typhi. PMID- 19679509 TI - Study on the interaction between methyl violet and bovine serum albumin by spectral analyses. AB - In this article the interaction between methyl violet (MV) and bovine serum albumin (BSA) was studied with spectroscopy. The results indicated that the fluorescence intensity of BSA was quenched strongly by MV through a static quenching procedure. The association constants, the number of binding sites and basic thermodynamic parameters were obtained based on fluorescence quenching data. The effect of MV on the conformation of BSA had been investigated with synchronous fluorescence spectroscopy and circular dichroism (CD) spectrum. PMID- 19679511 TI - Environmental adaptation of proteins: regression models with simple physicochemical properties. AB - Bio-sequences from ortholog proteins are well suited for statistical inference when the sequences can be divided into ordinal groups based on known environmental features or traits of the host organisms. In this paper two new regression models are described for extracting proteomic trends of extreme environments. The approach is based on physicochemical properties of the amino acids, and may also utilise stratification of the data. We are especially looking for connections of temperature adaptation between the organism and its molecular level. To show the applicability of the methods, we present analyses of genomic data from proteobacteria orders, where we examine the cold adaptation of membrane proteins, intracellular proteins, and the enzyme endonuclease I. Our results confirm earlier findings that redistribution of charge and increase of surface hydrophobicity might be some of the most important signatures for cold adaptation. PMID- 19679510 TI - Poor sleep and impaired self-care: towards a comprehensive model linking sleep, cognition, and heart failure outcomes. AB - BACKGROUND: Persons with heart failure (HF) have significantly lower sleep quantity and quality than persons without HF. The purpose of this article is to propose a conceptual model describing how poor sleep may contribute to inadequate self-care and untoward outcomes in persons with HF. AIMS: Our overarching hypothesis is that sleep affects self-care and outcomes through its effects on cognition. Building on the work of others, we outline a conceptual model that illustrates that even transient sleep disruption prevents sleep-related restorative processes and contributes to cognitive dysfunction-especially in the 25-50% of HF patients with existing cognitive impairment. Poor sleep may be sufficient to impair cognition to a level that interferes with higher order functions involved in effective HF self-care practices. Through these mechanisms, inadequate sleep may contribute to poor outcomes such as low health-related quality of life and greater risk of unplanned hospitalization. CONCLUSION: The proposed model (1) bridges physical, neuropsychological and behavioral phenomena, (2) suggests a mechanism by which poor sleep affects daytime behavior, and (3) is empirically testable. Exploring factors that interfere with sleep may improve self-care and outcomes in persons with HF. PMID- 19679512 TI - Computational analyses of mammalian lactate dehydrogenases: human, mouse, opossum and platypus LDHs. AB - Computational methods were used to predict the amino acid sequences and gene locations for mammalian lactate dehydrogenase (LDH) genes and proteins using genome sequence databanks. Human LDHA, LDHC and LDH6A genes were located in tandem on chromosome 11, while LDH6B and LDH6C genes were on chromosomes 15 and 12, respectively. Opossum LDHC and LDH6B genes were located in tandem with the opossum LDHA gene on chromosome 5 and contained 7 (LDHA and LDHC) or 8 (LDH6B) exons. An amino acid sequence prediction for the opossum LDH6B subunit gave an extended N-terminal sequence, similar to the human and mouse LDH6B sequences, which may support the export of this enzyme into mitochondria. The platypus genome contained at least 3 LDH genes encoding LDHA, LDHB and LDH6B subunits. Phylogenetic studies and sequence analyses indicated that LDHA, LDHB and LDH6B genes are present in all mammalian genomes examined, including a monotreme species (platypus), whereas the LDHC gene may have arisen more recently in marsupial mammals. PMID- 19679513 TI - ML or NJ-MCL? A comparison between two robust phylogenetic methods. AB - Large-scale gene sequencing gives an opportunity to reconstruct the tree of life and histories of multigene species phylogenies from very large datasets. A primary need for reconstructing large-scale phylogenies is a computationally efficient and accurate method. Current efforts to achieve such a goal include NJ MCL2 described by Tamura et al. (2004; 2007), an algorithm based on maximum likelihood (ML) and neighbor joining (NJ) algorithms. Although it has been reported that the NJ-MCL method performs better than the NJ method, studies comparing the accuracy of the ML and NJ-MCL methods are lacking. Here, accuracy of the NJ-MCL and the ML methods are examined. The concatenation approach (by progressive addition of genes) is used in a biologically realistic computer simulation to infer the accuracy of the methods. Simulation results clearly show that although NJ-MCL is computationally efficient and outperforms NJ method, the ML method is clearly much more accurate than the NJ-MCL method. The results encourage the use of the ML algorithm where datasets include up to several hundred species, but for reconstructing grand-scale phylogenies (i.e., where several thousand of taxa are included) NJ-MCL is preferred. PMID- 19679514 TI - Long-term upper and lower urinary tract functions in children with posterior urethral valves. AB - OBJECTIVE: Several factors have been identified as predictive of future renal function in children with posterior urethral valves (PUV). Our aim was to analyse upper and lower urinary tract outcome in patients with PUV, and determine any factors from the period of early management that were predictive of future renal function. METHODS: The charts of 67 boys (mean age 2.4 years) diagnosed with PUV were reviewed. The most common presenting symptom was dribbling in 43.2% and UTI in 28.3%. Twenty-three (34.8%) patients developed end-stage renal disease aged 1 15 years. The mean time of renal survival was calculated as 7.8 (SEM=0.73) years. RESULTS: Incontinence in patients over 5 years old, lower urinary tract dysfunction, serum creatinine level in first year or at the time of diagnosis, and presence of vesicoureteral reflux and high-grade bilateral reflux were significant risk factors for occurrence of renal failure in the future. Lower tract dysfunction was seen in 58.6% of patients. Postnatal diagnosis and presence of high-grade reflux were significant risk factors for the future occurrence of lower urinary tract dysfunction. CONCLUSION: It is important to recognize that PUV have consequences not only during childhood or before treatment, but also during or after the treatment period. Long-term assessment and follow-up of upper and lower urinary tract functions after valve ablation is necessary. PMID- 19679515 TI - Heminephroureterectomy for duplex kidney: laparoscopy versus open surgery. AB - OBJECTIVE: To report our experience of laparoscopic heminephroureterectomy (Hnu) in pediatric patients with duplex anomalies, in comparison to open surgery. PATIENTS AND METHODS: Retrospective review of data from patients who underwent Hnu from 2005 to 2008 was performed. The patients were divided into two groups: laparoscopic (LHnu) and open surgery (OHnu). Laparoscopic surgery was performed by transperitoneal approach in majority of cases. Open surgery was performed by retroperitoneal approach in all cases. RESULTS: Group LHnu: nine patients (8 females, 1 male) with median age of 14 months (range 3-205). Transperitoneal approach was performed in eight patients. Mean operative time was 182 min (CI 95% 146-217). No conversion to open surgery was necessary and there were no complications. Mean hospital stay was 2.44 days (CI 95% 1.37-3.52). Group OHnu: eight patients (3 females, 5 males) underwent nine heminephrectomies at median age of 6.9 months (range 1-12). Mean operating time was 152 min (CI 95% 121-183). There were no complications and mean hospital stay was 4.38 (CI 95% 2.59-6.16) days. Statistical analysis showed no statistically significant difference (P>0.05) in operating time between groups while mean hospital stay was significant (P=0.021). CONCLUSION: The laparoscopic approach is feasible, safe, reduces hospital stay, does not increase operating time and has better cosmetic results. We believe this should be the first option for heminephrectomy. PMID- 19679516 TI - Reliability and validity of a Chinese version of the Impact of Pediatric Epilepsy Scale. AB - The Impact of Pediatric Epilepsy Scale (IPES) is an accurate, acceptable, and quick tool that assesses the impact of epilepsy on the child with epilepsy and his or her family. The aim of this study was to investigate its applicability in China. After multistage translation and cultural adaptation, the final Chinese version was administered to 110 parents of children with epilepsy to evaluate its validity, reliability, and sensitivity. All items contributed significantly to the summary measure. With respect to validity, all items were substantially correlated with the criterion questionnaire subscales, and principal component analysis indicated that three factors accounted for 72% of the variance of the scale. The internal consistency coefficients of the first and second tests for the total were 0.916 and 0.930, respectively, and test-retest reliability ranged from 0.891 to 0.992. Additionally, the IPES can detect differences in health related quality of life (HRQOL) between subjects according to epilepsy severity. In conclusion, this study indicates that the Chinese IPES has good validity, reliability, and sensitivity, and is an epilepsy-specific HRQOL questionnaire that is a brief, accurate, and valid assessment of the influence of epilepsy on the child and family. PMID- 19679517 TI - Differential control of collagen synthesis by the sympathetic and renin angiotensin systems in the rat left ventricle. AB - In the present study, we tested the hypothesis of the indirect (via the sympathetic nervous system (SNS)) and direct (via AT1 receptors) contributions of Angiotensin II (Ang II) on the synthesis of collagen types I and III in the left ventricle (LV) in vivo. Sympathectomy and blockade of the Ang II receptor AT1 were performed alone or in combination in normotensive rats. The mRNA and protein synthesis of collagen types I and III were examined by Q-RT-PCR and immunoblotting in the LV. Collagen types I and III mRNA were decreased respectively by 53% and 22% after sympathectomy and only collagen type I mRNA was increased by 52% after AT1 receptor blockade. mRNA was not changed for collagen type I but was decreased by 25% for collagen type III after double treatment. Only collagen protein type III was decreased after sympathectomy by 12%, but collagen proteins were increased respectively for types I and III by 145% and 52% after AT1 receptor blockade and by 45% and 60% after double treatment. Deducted interpretations from our experimental approach suggest that Ang II stimulates indirectly (via SNS) and inhibits directly (via AT1 receptors) the collagen type I at transcriptional and protein levels. For collagen type III, it stimulates indirectly the transcription and inhibited directly the protein level. Therefore, the Ang II regulates collagen synthesis differently through indirect and direct pathways. PMID- 19679519 TI - Secular trend in height in Al Ain-United Arab Emirates. AB - Correlation between cycles in human stature and those in economic variables is well established. A recent review of international trends in this area provided information from most parts of the world but none on Arabs in the Middle East or more specifically the gulf region. The United Arab Emirates experienced a transformation in economic and social life followed the discovery of oil in the late 1960s and the wealth that it generated. No data is available on human growth at this period of time because its population never had health services prior to the 1970s. A study on conventional cardiovascular risk factors in 2004-2005 included 817 randomly selected national adults (> or =18 years) from both genders. The relationship between height and age in this study showed both men and women have increased in height with time demonstrating the secular change in height most likely a result of changing socioeconomic factors. PMID- 19679520 TI - Dynamics of celiac disease-specific serology after initiation of a gluten-free diet and use in the assessment of compliance with treatment. AB - BACKGROUND: The usefulness of celiac disease-related serology in monitoring patients on a gluten-free diet has been debated. AIM: To describe serologic changes over time and assess whether serology tests can predict compliance with the gluten-free diet. METHODS: Sera obtained at baseline and every 3 months thereafter for 1 year in 82 adult celiac disease patients were assayed for: (1) IgA antigliadin, (2) IgA anti-tissue transglutaminase, (3) IgA endomysial, (4) IgA, and (5) IgG anti-deamidated gliadin peptides, (6) dual detection of IgA and IgG anti-deamidated gliadin peptides, (7) a single assay for IgA and IgG of both anti-deamidated gliadin peptide and anti-tissue transglutaminase, and (8) IgA antiactin antibodies. RESULTS: At 3 months after diagnosis, most antibody assays significant decrease in mean concentrations (p<0.0001) and the percentage of positive samples (p<0.0001) with further improvement in subsequent determinations. Strictly adherents had significantly lower concentrations of antibodies (p<0.01 to p<0.00001) and smaller proportion of positive samples for IgA endomysial, IgA antiactin antibodies and IgA antigliadin (15.6%, 17.4% and 23.9%, respectively) than partially compliant. At 1 year, IgA endomysial (p<0.02), IgA antiactin antibodies (p<0.05) and anti-tissue transglutaminase (p<0.02) predicted the degree of compliance. CONCLUSIONS: Gluten-free diet treatment produced rapid and significant qualitative and quantitative changes in celiac disease-related antibodies which may be useful for monitoring dietary compliance. PMID- 19679521 TI - A comparison of actual-ideal weight discrepancy, body appreciation, and media influence between street-dancers and non-dancers. AB - Previous work on body image has tended to treat dancers as a relatively homogenous group, despite the existence of different dance styles and genres. In the current study, we examined body image among individuals involved in street dancing (genres that typically evolved outside formal settings and are often improvisational in nature) and an age-matched comparison of non-dancers. A total of 83 street-dancers and 84 non-dancers completed scales measuring their actual ideal weight discrepancy, body appreciation, sociocultural attitudes toward appearance, and demographics. Controlling for participant body mass index (BMI), results showed no significant between-group difference in actual-ideal weight discrepancy, although street-dancers had significantly higher body appreciation than non-dancers. In addition, media influences were implicated in body image concepts for both groups, although internalisation of athletic ideals was more important for street-dancers. These results are discussed in relation to the extant research on body image among dancers. PMID- 19679522 TI - Outcome analysis of cleft palate surgery--can static 2D MRI replace videofluroscopy? PMID- 19679518 TI - Visceral organ cross-sensitization - an integrated perspective. AB - Viscero-somatic referral and sensitization has been well documented clinically and widely investigated, whereas viscero-visceral referral and sensitization (termed cross-organ sensitization) has only recently received attention as important to visceral disease states. Because second order neurons in the CNS have been extensively shown to receive convergent input from different visceral organs, it has been assumed that cross-organ sensitization arises by the same convergence-projection mechanism as advanced for viscero-somatic referral and sensitization. However, increasing evidence also suggests participation of peripheral mechanisms to explain referral and sensitization. We briefly summarize behavioral, morphological and physiological support of and focus on potential mechanisms underlying cross-organ sensitization. PMID- 19679523 TI - Influence of decantation, washing and centrifugation on adipocyte and mesenchymal stem cell content of aspirated adipose tissue: a comparative study. AB - BACKGROUND: In the last decade, controversy has arisen regarding the influence of fat harvesting, processing and injection techniques on adipose tissue graft. The aim of this study is to compare the influence of three widely used fat processing techniques in plastic surgery on the viability and number of adipocytes and mesenchymal stem cells (MSCs) of aspirated fat. METHODS: A prospective cross sectional study was conducted in 20 adult healthy female patients in whom material obtained by liposuction of the lower abdomen was separated and processed by decantation, washing or centrifugation. The morphology and quantity of adipocytes were determined by histological analysis. The viability and number of MSCs in the middle layer of each lipoaspirate and the pellet derived from centrifuged samples were obtained by multi-colour flow cytometry. RESULTS: Cell count per high-powered field of intact nucleated adipocytes was significantly greater in decanted lipoaspirates, whereas centrifuged samples showed a greater majority of altered adipocytes. MSC concentration was significantly higher in washed lipoaspirates compared to decanted and centrifuged samples. However, the pellet collected at the bottom of the centrifuged samples showed the highest concentration of MSCs. CONCLUSION: Based on the theory of cell survival stating the importance of adipocytes' integrity for graft survival and the theory claiming the importance of regenerative MSCs in the maintenance and stabilisation of fat transplant, washing may turn out to be the best processing technique for adipose tissue graft take. While eliminating most contaminants during the process, it preserved and maintained the quantity, integrity and viability of the most important components of aspirated adipose tissue. PMID- 19679525 TI - [Janos Balassa memorial lecture]. PMID- 19679524 TI - Color-coding improves parental understanding of body mass index charting. AB - OBJECTIVE: To assess parental understanding of body mass index (BMI) and BMI percentiles by using standard versus color-coded charts; to investigate how parental literacy and/or numeracy (quantitative skills) affects that understanding. METHODS: A convenience sample of 163 parents of children aged 2 to 8 years at 2 academic pediatric centers completed a demographics questionnaire, the mathematics portion of the Wide Range Achievement Test (WRAT-3R), the Short Test of Functional Health Literacy in Adults (S-TOFHLA), and an "Understanding BMI" questionnaire, which included parallel BMI charting questions to compare understanding of standard versus color-coded BMI charting. Outcomes included parental-reported versus actual understanding of BMI, the odds (obtained by generalized estimating equations) of answering parallel questions correctly by using standard versus color-coded charting, and odds of answering questions correctly on the basis of numeracy and literacy. RESULTS: Many parents (60%) reported knowing what BMI was, but only 30% could define it even roughly correctly. When parents used color-coded charts, they had greater odds of answering parallel BMI charting questions correctly than when they used standard charts (mean, 88% vs 65% correct; pooled adjusted odds ratio, 4.32; 95% confidence interval, 3.14-5.95; P < .01). Additionally, parents with lower numeracy (K-5 level) benefited more from color-coded charts (increased from 51% to 81% correct) than did higher numeracy parents (high school level or greater), who performed well with both charts (89% vs 99% correct). CONCLUSIONS: Parents consistently performed better with color-coded than standard BMI charts. Color coding was particularly helpful for lower numeracy parents. Future studies should investigate whether these results translate into the office setting and whether understanding motivates parents to implement important lifestyle changes. PMID- 19679526 TI - [Effect of postconditioning in major vascular operations on rats]. AB - BACKGROUND: Postconditioning - using alternating brief cycles of reperfusion/reocclusion applied just at the very beginning of reperfusion - has recently been described as a potent therapeutic technique, attenuating ischaemia reperfusion injury. In vascular surgery, certain elective interventions involve cross-clamping of major arteries, resulting in temporary ischaemia in large peripheral organs, which thus suffer ischaemia-reperfusion injury. Patients undergoing these operations may develop also serious systemic complications such as multiple distant organ dysfunctions, SIRS, detrimental redistribution of the circulation or even shock, a phenomenon called reperfusion-syndrome. We studied the effects of postconditioning on reperfusion-syndrome in a rodent experimental model. MATERIAL AND METHODS: Anaesthetized male Wistar rats underwent 180 minutes of bilateral lower limb ischaemia and 4 hours of reperfusion using an infrarenal cross-clamping of the abdominal aorta. Control animals underwent no additional intervention. Postconditioning consisted of 6 cycles of 10-second aortic occlusion/10-second declamping starting at the beginning of reperfusion. Haemodynamic parameters were observed with invasive arterial manometer, microcirculation of the lower limb was detected with laser-Doppler-flowmeter. After 4 hours of reperfusion serum, urine, and histological samples were collected. RESULTS: 180-minute ischaemia resulted in significant haemodynamic changes after reperfusion. Postconditioning affected the character of the microcirculatory flow curves, the limb circulation stabilized with hyperaemia after reperfusion. Postconditioning caused a significant reduction in systemic inflammatory response (TNF-alpha, oxygen-derived free radicals). The laboratory and histological samples implied a significant decrease in remote organ (lung and renal) dysfunctions after postconditioning. CONCLUSION: Postconditioning proves to be capable in conferring protection against different organ injuries caused by longer circulatory occlusions during elective major vascular surgeries. PMID- 19679528 TI - [Fewer German surgeons--new trends in surgical training]. PMID- 19679527 TI - [Clinical value of "zero-hour biopsy"]. AB - In spite of the improving results the long-term benefits of renal transplantation fell behind the expectable potential possibilities. The main cause of kidney graft loss is chronic allograft nephropathy following cardiovascular deaths. This is such a multiple etiologic clinical picture which may occur at any time. When adequate treatment is not available in time repeated development of chronic renal failure is unavoidable. The aim of my study is, how can we rise the number of transplanted kidney and the quality of them. PMID- 19679529 TI - [Surgery of oesophagus]. PMID- 19679530 TI - [Gastric surgery]. PMID- 19679531 TI - [Current surgical treatment of IBD]. PMID- 19679532 TI - [Surgical problems of anal incontinence]. PMID- 19679533 TI - [Oncologic surgery of the intestines]. PMID- 19679534 TI - [Liver surgery]. PMID- 19679535 TI - [Liver transplantation]. PMID- 19679536 TI - [Biliary surgery]. PMID- 19679537 TI - [Surgery of the pancreas]. PMID- 19679538 TI - [Abdominal catastrophe]. PMID- 19679539 TI - [NOTES (Natural Orifice Translumenal Endoscopic Surgery)]. PMID- 19679542 TI - Tracking lexical access in speech production: electrophysiological correlates of word frequency and cognate effects. AB - The present study establishes an electrophysiological index of lexical access in speech production by exploring the locus of the frequency and cognate effects during overt naming. We conducted 2 event-related potential (ERP) studies with 16 Spanish-Catalan bilinguals performing a picture naming task in Spanish (L1) and 16 Catalan-Spanish bilinguals performing a picture naming task in Spanish (L2). Behavioral results showed a clear frequency effect and an interaction between frequency and cognate status. The ERP elicited during the production of high frequency words diverged from the low-frequency ERP between 150 and 200 ms post target presentation and kept diverging until voice onset. The same results were obtained when comparing cognate and noncognate conditions. Positive correlations were observed between naming latencies and mean amplitude of the P2 component following the divergence, for both the lexical frequency and the cognate effects. We conclude that lexical access during picture naming begins approximately 180 ms after picture presentation. Furthermore, these results offer direct electrophysiological evidence for an early influence of frequency and cognate status in speech production. The theoretical implications of these findings for models of speech production are discussed. PMID- 19679543 TI - Uncertainty during anticipation modulates neural responses to aversion in human insula and amygdala. AB - Uncertainty about potential negative future outcomes can cause stress and is a central feature of anxiety disorders. The stress and anxiety associated with uncertain situations may lead individuals to overestimate the frequency with which uncertain cues are followed by negative outcomes, an example of covariation bias. Using functional magnetic resonance imaging, we found that uncertainty related expectations modulated neural responses to aversion. Insula and amygdala responses to aversive pictures were larger after an uncertain cue (that preceded aversive or neutral pictures) than a certain cue (that always preceded aversive pictures). Anticipatory anterior cingulate cortex (ACC) activity elicited by the cues was inversely associated with the insula and amygdala responses to aversive pictures following the cues. Nearly 75% of subjects overestimated the frequency of aversive pictures following uncertain cues, and ACC and insula activity predicted this uncertainty-related covariation bias. Findings provide the first evidence of the brain mechanisms of covariation bias and highlight the temporal dynamics of ACC, insula, and amygdala recruitment for processing aversion in the context of uncertainty. PMID- 19679544 TI - The effect of variation in expression of the candidate dyslexia susceptibility gene homolog Kiaa0319 on neuronal migration and dendritic morphology in the rat. AB - We investigated the postnatal effects of embryonic knockdown and overexpression of the candidate dyslexia gene homolog Kiaa0319. We used in utero electroporation to transfect cells in E15/16 rat neocortical ventricular zone with either 1) small hairpin RNA (shRNA) vectors targeting Kiaa0319, 2) a KIAA0319 expression construct, 3) Kiaa0319 shRNA along with KIAA0319 expression construct ("rescue"), or 4) a scrambled version of Kiaa0319 shRNA. Knockdown, but not overexpression, of Kiaa0319 resulted in periventricular heterotopias that contained large numbers of both transfected and non-transfected neurons. This suggested that Kiaa0319 shRNA disrupts neuronal migration by cell autonomous as well as non-cell autonomous mechanisms. Of the Kiaa0319 shRNA-transfected neurons that migrated into the cortical plate, most migrated to their appropriate lamina. In contrast, neurons transfected with the KIAA0319 expression vector attained laminar positions subjacent to their expected positions. Neurons transfected with Kiaa0319 shRNA exhibited apical, but not basal, dendrite hypertrophy, which was rescued by overexpression of KIAA0319. The results provide additional supportive evidence linking candidate dyslexia susceptibility genes to migrational disturbances during brain development, and extends the role of Kiaa0319 to include growth and differentiation of dendrites. PMID- 19679545 TI - Melanoma proliferation and chemoresistance controlled by the DEK oncogene. AB - Gain of chromosome 6p is a consistent feature of advanced melanomas. However, the identity of putative oncogene(s) associated with this amplification has remained elusive. The chromatin remodeling factor DEK is an attractive candidate as it maps to 6p (within common melanoma-amplified loci). Moreover, DEK expression is increased in metastatic melanomas, although the functional relevance of this induction remains unclear. Importantly, in other tumor types, DEK can display various tumorigenic effects in part through its ability to promote proliferation and inhibit p53-dependent apoptosis. Here, we report a generalized up-regulation of DEK protein in aggressive melanoma cells and tumors. In addition, we provide genetic and mechanistic evidence to support a key role of DEK in the maintenance of malignant phenotypes of melanoma cells. Specifically, we show that long-term DEK down-regulation by independent short hairpin RNAs resulted in premature senescence of a variety of melanoma cell lines. Short-term abrogation of DEK expression was also functionally relevant, as it attenuated the traditional resistance of melanomas to DNA-damaging agents. Unexpectedly, DEK short hairpin RNA had no effect on p53 levels or p53-dependent apoptosis. Instead, we identified a new role for DEK in the transcriptional activation of the antiapoptotic MCL-1. Other MCL-1-related factors such as BCL-2 or BCL-xL were unaffected by changes in the endogenous levels of DEK, indicating a selective effect of this gene on the apoptotic machinery of melanoma cells. These results provide support for DEK as a long sought-after oncogene mapping at chromosome 6, with novel functions in melanoma proliferation and chemoresistance. PMID- 19679547 TI - Premenopausal mammographic density in relation to cyclic variations in endogenous sex hormone levels, prolactin, and insulin-like growth factors. AB - Mammographic density is strongly associated with breast cancer risk, and endogenous hormones, which are risk factors for breast cancer, may be involved in the mechanism. This cross-sectional study of 494 premenopausal women is the first to account for cyclic variations in estrogen levels, by measuring urinary estrone glucuronide (E1G) in the periovulatory and luteal phases of the menstrual cycle, and to assess the role of androgens. Computer-assisted density readings were obtained from digitized mammograms. Mean ovulatory E1G level and daily E1G load were both positively associated with percent density before adjustment for body mass index (BMI), with women in the top fourth having 10.2% (95% CI: 2.9%, 18.1%) and 8.9% (1.7%, 16.7%), respectively, higher density than those in the bottom fourth (Ptrend before/after BMI adjustment=0.006/0.11 and 0.01/0.13, respectively). Neither the peak nor luteal E1G levels were predictive of density after adjustment for E1G levels at other points in the cycle. The plasma androgens testosterone, androstenedione, and dehydroepiandrosterone sulfate were negatively associated with density. In mutually adjusted analyses, density was positively associated with insulin-like growth factor (IGF)-I and negatively with IGF-II (Ptrend=0.006 for both) but not with IGF binding protein-3. There was also weak evidence of a positive association of prolactin with density. The study supports the hypothesis that endogenous hormones affect density in premenopausal women; in particular, it shows a positive association between estrogen levels and density and suggests that the mean level throughout the cycle is the most biologically relevant measure. Most of these hormone-density associations were attenuated with further adjustment for BMI. PMID- 19679546 TI - Cell type-specific targeted mutations of Kras and Pten document proliferation arrest in granulosa cells versus oncogenic insult to ovarian surface epithelial cells. AB - The small G-protein KRAS is crucial for mediating gonadotropin-induced events associated with ovulation. However, constitutive expression of KrasG12D in granulosa cells disrupted normal follicle development leading to the persistence of abnormal follicle-like structures containing nonmitotic cells. To determine what factors mediate this potent effect of KrasG12D, gene profiling analyses were done. We also analyzed KrasG12D;Cyp19-Cre and KrasG12;Pgr-Cre mutant mouse models that express Cre prior to or after the initiation of granulosa cell differentiation, respectively. KrasG12D induced cell cycle arrest in granulosa cells of the KrasG12D;Cyp19-Cre mice but not in the KrasG12D;Pgr-Cre mice, documenting the cell context-specific effect of KrasG12D. Expression of KrasG12D silenced the Kras gene, reduced cell cycle activator genes, and impaired the expression of granulosa cell and oocyte-specific genes. Conversely, levels of PTEN and phosphorylated p38 mitogen-activated protein kinase (MAPK) increased markedly in the mutant granulosa cells. Because disrupting Pten in granulosa cells leads to increased proliferation and survival, Pten was disrupted in the KrasG12D mutant mice. The Pten/Kras mutant mice were infertile but lacked granulosa cell tumors. By contrast, the Ptenfl/fl;KrasG12D;Amhr2-Cre mice developed aggressive ovarian surface epithelial cell tumors that did not occur in the Ptenfl/fl;KrasG12D;Cyp19-Cre or Ptenfl/fl;KrasG12D;Pgr-Cre mouse strains. These data document unequivocally that Amhr2-Cre is expressed in and mediates allelic recombination of oncogenic genes in ovarian surface epithelial cells. That KrasG12D/Pten mutant granulosa cells do not transform but rather undergo cell cycle arrest indicates that they resist the oncogenic insults of Kras/Pten by robust self-protecting mechanisms that silence the Kras gene and elevate PTEN and phosphorylated p38 MAPK. PMID- 19679548 TI - The decline in U.S. cancer mortality in people born since 1925. AB - The conventional practice of analyzing overall age-adjusted cancer mortality rates heavily emphasizes the experience of older, higher mortality age groups. This may conceal shifts in lifetime cancer mortality experience emerging first in younger age groups. We examined age-specific cancer mortality rates and birth cohort-specific cancer mortality rates in U.S. mortality data recorded since 1955 to assess the effects of age, period, and cohort in secular mortality trends. Cancer mortality and population data were obtained from WHO Statistical Information System. Age-specific cancer mortality rates have been steadily declining in the United States since the early 1950s, beginning with children and young adults and now including all age groups. During the second half of the 20th century, each successive decade of births from 1925 to 1995 experienced a lower risk of cancer death than its predecessor at virtually every age for which such a comparison can be made. A major decline in cancer mortality has been occurring in the United States for the past 50 years, affecting birth cohorts born as long as 80 years ago. Excepting lung cancer, much of this decline has occurred despite relatively stable cancer incidence. These findings suggest that improvements in cancer detection, treatment, and/or prevention have reduced the risk of cancer death across the life span for individuals born in the last three quarters of the 20th century. PMID- 19679549 TI - Metformin disrupts crosstalk between G protein-coupled receptor and insulin receptor signaling systems and inhibits pancreatic cancer growth. AB - Recently, we identified a novel crosstalk between insulin and G protein-coupled receptor (GPCR) signaling pathways in human pancreatic cancer cells. Insulin enhanced GPCR signaling through a rapamycin-sensitive mTOR-dependent pathway. Metformin, the most widely used drug in the treatment of type 2 diabetes, activates AMP kinase (AMPK), which negatively regulates mTOR. Here, we determined whether metformin disrupts the crosstalk between insulin receptor and GPCR signaling in pancreatic cancer cells. Treatment of human pancreatic cancer cells (PANC-1, MIAPaCa-2, and BxPC-3) with insulin (10 ng/mL) for 5 minutes markedly enhanced the increase in intracellular [Ca(2+)] induced by GPCR agonists (e.g., neurotensin, bradykinin, and angiotensin II). Metformin pretreatment completely abrogated insulin-induced potentiation of Ca(2+) signaling but did not interfere with the effect of GPCR agonists alone. Insulin also enhanced GPCR agonist induced growth, measured by DNA synthesis, and the number of cells cultured in adherent or nonadherent conditions. Low doses of metformin (0.1-0.5 mmol/L) blocked the stimulation of DNA synthesis, and the anchorage-dependent and anchorage-independent growth induced by insulin and GPCR agonists. Treatment with metformin induced striking and sustained increase in the phosphorylation of AMPK at Thr(172) and a selective AMPK inhibitor (compound C, at 5 micromol/L) reversed the effects of metformin on [Ca(2+)](i) and DNA synthesis, indicating that metformin acts through AMPK activation. In view of these results, we tested whether metformin inhibits pancreatic cancer growth. Administration of metformin significantly decreased the growth of MIAPaCa-2 and PANC-1 cells xenografted on the flank of nude mice. These results raise the possibility that metformin could be a potential candidate in novel treatment strategies for human pancreatic cancer. PMID- 19679551 TI - The dendritic cell-like functions of IFN-producing killer dendritic cells reside in the CD11b+ subset and are licensed by tumor cells. AB - IFN producing killer dendritic cells (IKDC) were originally defined as CD11c(int) B220(+)NK1.1(+) (or CD49b(+)) cells that exert a potent tumoricidal activity in animals lacking B, T, and conventional natural killer effectors. MHC class II expression on tumor infiltrating IKDC prompted us to investigate their putative antigen presenting function. Here, we show that tumor cells license IKDC to acquire the properties of antigen presenting cells, i.e., expression of MHC class II and costimulatory CD86 molecules. We show that the CD11b(+) subset of IKDC are able to prime naive CD4(+) T cells and cross-prime naive CD8(+) T lymphocytes. Licensing of IKDC by tumor cells was mandatory for the full differentiation of T cells into polarized effectors. IKDC could engulf and process soluble Ova protein in a CD206-dependent manner. Finally, we show that CD11b(+)IKDC is selectively endowed with CTLA4Ig-inhibitable antigen presenting capacities and that targeting this subset with the detoxified adenylate cyclase toxin of Bordetella pertussis fused to antigen resulted in efficient cross-presentation of antigen by IKDC to specific TCR transgenic CD8(+)T cells in vivo. Collectively, our data indicate that upon exposure to tumor cells, IKDC subserve DC-like functions. PMID- 19679550 TI - Cardiac glycosides inhibit p53 synthesis by a mechanism relieved by Src or MAPK inhibition. AB - p53 is regulated at multiple levels. We report here that p53, in multiple lines of human cancer cells, is down-regulated by cardiac glycoside drugs digoxin and ouabain, potent inhibitors of Na(+)/K(+)-ATPase. These drugs reduced the basal levels of p53 protein at nanomolar concentrations in a dose-, time-, and cancer cell line-dependent manner, but independent of p53 status of wild-type or mutant. The drugs also reduced the levels of p53 induced by its activators as well as p53 transfected into human cancer cells, regardless of its status. Interestingly, the drugs had no effect on endogenous p53 in two immortalized human cell lines. Mechanistically, p53 reduction occurred not at the mRNA levels but at the protein levels, as a result of reduced protein synthesis rather than enhanced degradation. The cellular sensitivity to drug-induced p53 reduction was not associated with the levels of alphasubunits of Na(+)/K(+)-ATPase in different cell lines. Although lowering extracellular K(+) did not reduce p53 as did ouabain and digoxin, it did potentiate both digoxin- and ouabain-induced p53 reduction in sensitive lines. Finally, p53 reduction seems to be triggered by activation of Src/mitogen-activated protein kinase (MAPK) signaling pathways upon drug binding to the Na(+)/K(+)-ATPase and can be completely blocked by the inhibitors of Src or MAP/ERK kinase. This is the first report that cardiac glycoside drugs, by initiating the Src/MAPK signaling pathways, reduce the p53 levels via inhibition of p53 protein synthesis. The drugs may be useful in the treatment of human cancers with a gain-of-function p53 mutation. PMID- 19679552 TI - IFN-producing killer dendritic cells are antigen-presenting cells endowed with T cell cross-priming capacity. AB - IFN-producing killer dendritic cells (IKDC) represent a recently discovered cell type in the immune system that possesses a number of functions contributing to innate and adaptive immunity, including production of type 1 and 2 IFNs, interleukin (IL)-12, natural killing, and ultimately antigen presentation to naive T cells. Here, we compared in vitro and in vivo responses of mouse IKDC, conventional dendritic cells (DC), and natural killer (NK) cells to murine cytomegalovirus infection and found distinct functions among these cell subsets. Upon recognition of infected fibroblasts, IKDC, as well as NK, produced high level of IFN-gamma, but unlike NK, IKDC simultaneously produced IL-12p40 and up regulated MHC class II (MHC-II) and costimulatory molecules. Using MHC-II molecule expression as a phenotypic marker to distinguish activated IKDC from activated NK, we further showed that highly purified MHC-II(+) IKDC but not NK cross-present MHC class I-restricted antigens derived from MCMV-infected targets to CD8(+) T cells in vitro and in vivo. Our findings emphasize the unique nature of IKDC as a killer antigen-presenting cell directly linking innate and adaptive immunity. PMID- 19679553 TI - Cullin 1 functions as a centrosomal suppressor of centriole multiplication by regulating polo-like kinase 4 protein levels. AB - Abnormal centrosome and centriole numbers are frequently detected in tumor cells where they can contribute to mitotic aberrations that cause chromosome missegregation and aneuploidy. The molecular mechanisms of centriole overduplication in malignant cells, however, are poorly characterized. Here, we show that the core SKP1-cullin-F-box component cullin 1 (CUL1) localizes to maternal centrioles and that CUL1 is critical for suppressing centriole overduplication through multiplication, a recently discovered mechanism whereby multiple daughter centrioles form concurrently at single maternal centrioles. We found that this activity of CUL1 involves the degradation of Polo-like kinase 4 (PLK4) at maternal centrioles. PLK4 is required for centriole duplication and strongly stimulates centriole multiplication when aberrantly expressed. We found that CUL1 is critical for the degradation of active PLK4 following deregulation of cyclin E/cyclin-dependent kinase 2 activity, as is frequently observed in human cancer cells, as well as for baseline PLK4 protein stability. Collectively, our results suggest that CUL1 may function as a tumor suppressor by regulating PLK4 protein levels and thereby restraining excessive daughter centriole formation at maternal centrioles. PMID- 19679554 TI - Identification of nectin-4 oncoprotein as a diagnostic and therapeutic target for lung cancer. AB - Gene expression profile analysis of lung cancers revealed the transactivation of an immunoglobulin-like molecule Nectin-4 in the majority of non-small cell lung cancers (NSCLC). Immunohistochemical staining of 422 NSCLCs showed that a high level of Nectin-4 expression was associated with poor prognosis for NSCLC patients (P < 0.0001), and multivariate analysis confirmed its independent prognostic value (P < 0.0001). We established an ELISA to measure serum Nectin-4 and found that serum Nectin-4 levels were significantly higher in NSCLC patients than in healthy volunteers. The proportion of the serum Nectin-4-positive cases was 88 of 164 (53.7%) NSCLCs, whereas only 3 of 131 (2.3%) healthy volunteers were falsely diagnosed as positive, which was superior to carcinoembryonic antigen (CEA) and cytokeratin 19-fragment (CYFRA21-1) in sensitivity and specificity. A combined ELISA for both Nectin-4 and CEA increased sensitivity and classified 65.0% of lung adenocarcinomas as positive with false-positive rate of 4.6%. The use of both Nectin-4 and CYFRA21-1 classified 68.3% of lung squamous cell carcinomas as positive with false-positive rate of 6.1%. Treatment of lung cancer cells with small interfering RNAs against Nectin-4 suppressed its expression and cell growth. In addition, exogenous expression of Nectin-4 increased the lamellipodia formation and the invasive ability of mammalian cells through activation of small GTPase Rac1. Nectin-4 might play a significant role in lung carcinogenesis, and it should be a new candidate serum and tissue biomarker, as well as a therapeutic target. PMID- 19679556 TI - Osteoclast-derived matrix metalloproteinase-7, but not matrix metalloproteinase 9, contributes to tumor-induced osteolysis. AB - The matrix metalloproteinases MMP-2, MMP-3, MMP-7, MMP-9, and MMP-13 are highly expressed in the tumor-bone microenvironment, and, of these, MMP-7 and MMP-9 were found to be localized to bone-resorbing osteoclasts in human breast-to-bone metastases. In a bid to define the roles of host-derived MMP-7 and MMP-9 in the tumor-bone microenvironment, the tibias of MMP-7 and MMP-9 null mice were injected with osteolytic luciferase-tagged mammary tumor cell lines. Our data show that osteoclast-derived MMP-7 significantly contributes to tumor growth and tumor-induced osteolysis whereas osteoclast-derived MMP-9 had no effect on these processes. MMP-7 is capable of processing a number of nonmatrix molecules to soluble active forms that have profound effects on cell-cell communication, such as RANKL, a crucial mediator of osteoclast precursor recruitment and maturation. Therefore, the ability of osteoclast-derived MMP-7 to promote RANKL solubilization in the tumor-bone microenvironment was explored. Results revealed that levels of soluble RANKL were significantly lower in the MMP-7 null mice compared with wild-type (WT) controls. In keeping with this observation, MMP-7 null mice had significantly fewer osteoclast numbers at the tumor-bone interface compared with the WT controls. In summary, we propose that the solubilization of RANKL by MMP-7 is a potential mechanism through which MMP-7 mediates mammary tumor-induced osteolysis. Our studies indicate that the selective inhibition of MMP-7 in the tumor-bone microenvironment may be of benefit for the treatment of lytic breast-to-bone metastases. PMID- 19679555 TI - Overexpression of protease-activated receptor-1 contributes to melanoma metastasis via regulation of connexin 43. AB - Protease-activated receptor-1 (PAR-1) is a key player in melanoma metastasis with higher expression seen in metastatic melanoma cell lines and tissue specimens. cDNA microarray and Western blot analyses reveal that the gap junctional intracellular communication molecule connexin 43 (Cx-43), known to be involved in tumor cell diapedesis and attachment to endothelial cells, is significantly decreased after PAR-1 silencing in metastatic melanoma cell lines. Furthermore, Cx-43 promoter activity was significantly inhibited in PAR-1-silenced cells, suggesting that PAR-1 regulates Cx-43 at the transcriptional level. Chromatin immunoprecipitation studies showed a reduction in the binding of SP-1 and AP-1 transcription factors to the promoter of Cx-43. Both transcription factors have been shown previously to be required for maximal Cx-43 promoter activity. These results were corroborated by mutating the AP-1 and SP-1 binding sites resulting in decreased Cx-43 promoter activity in PAR-1-positive cells. Moreover, as Cx-43 has been shown to facilitate arrest of circulating tumor cells at the vascular endothelium, melanoma cell attachment to endothelial cells was significantly decreased in PAR-1-silenced cells, with this effect being abrogated after PAR-1 rescue. Herein, we report that up-regulation of PAR-1 expression, seen in melanoma progression, mediates high levels of Cx-43 expression. As both SP-1 and AP-1 transcription factors act as positive regulators of Cx-43, our data provide a novel mechanism for the regulation of Cx-43 expression by PAR-1. Indeed, Cx-43 expression was restored following PAR-1 rescue in PAR-1-silenced cells. Taken together, our data support the tumor promoting function of Cx-43 in melanoma. PMID- 19679557 TI - Mupirocin for preventing exit-site infection and peritonitis in patients undergoing peritoneal dialysis. AB - OBJECTIVES: Recently, there have been increasing concerns about the emergence of mupirocin resistance and increased infections due to lowered inhibition of Staphylococcus aureus. We conducted this systemic analysis to find out whether the application of mupirocin was effective for the prevention of exit-site infection (ESI) and peritonitis in patients undergoing peritoneal dialysis (PD). METHODS: Recruited studies met the following criteria: they were randomized controlled trials or historical cohort studies; subjects consisted of adults (age, >or= 18 years) undergoing PD; mupirocin treatment was administered to the therapy group and placebo or no treatment was administered to the control group. The primary extracted data were the difference in the episodes of ESI and peritonitis S. aureus or other organisms among treatment and control groups. Results. Fourteen studies described in 13 articles and a total of 1,233 patients versus 1,217 controls were included in the analysis. Of the 13 articles, 6 were newly published articles that had not been analysed previously and 3 were randomized controlled trials. The application of mupirocin decreased the risk by 72% [95% confidence interval (CI): 0.60-0.81] in ESI and by 70% (95% CI 0.52 0.81) in peritonitis due to S. aureus among all patients undergoing PD. Treatment of mupirocin reduced the risks of ESI and peritonitis due to all organisms by 57% (95% CI: 0.46-0.66) and 41% (95% CI: 0.24-0.54), respectively. Based on the six newly published articles, the reduced risk rate for mupirocin therapy was found to be 80% (95% CI: 0.39-0.93, P = 0.004) in ESI and 91% (95% CI: 0.72-0.97, P < 0.0001) in peritonitis due to S. aureus; 70% (95% CI: 0.47-0.82, P < 0.0001) in ESI and 42% (95% CI: 0.25-0.55, P < 0.0001) in peritonitis due to all organisms among mupirocin-treated and -untreated subjects. Based on the three randomized controlled trials, ESI and peritonitis due to S. aureus were found to be reduced by 73% (95% CI: 0.63-0.80, P < 0.0001) and 40% (95% CI: 0.17-0.56, P = 0.002), respectively. Interestingly, although mupirocin treatment can reduce the risk rate of ESI by 46% (95% CI: 0.35-0.55, P < 0.00001), it cannot decrease the risk rate of peritonitis due to all organisms (P = 0.56). CONCLUSIONS: Mupirocin prophylaxis was effective on preventing ESI and peritonitis due to S. aureus and other organisms in PD patients. PMID- 19679559 TI - NT-proBNP, fluid volume overload and dialysis modality are independent predictors of mortality in ESRD patients. AB - BACKGROUND: N-terminal fragment of B-type natriuretic peptide (NT-proBNP) is a marker of both fluid volume overload and myocardial damage, and it has been useful as a predictor of mortality in patients with end-stage renal disease (ESRD). It has been suggested that continuous ambulatory peritoneal dialysis (CAPD), automated peritoneal dialysis (APD) and haemodialysis (HD) may have different effects on fluid volume and blood pressure control; however, whether the independent predictive value of NT-proBNP for mortality is preserved when analysed in conjunction with fluid overload and dialysis modality is not clear. METHODS: A prospective multicentre cohort of 753 prevalent adult patients on CAPD, APD and HD was followed up for 16 months. Plasmatic levels of NT-proBNP, extracellular fluid volume/total body water ratio (ECFv/TBW) and traditional clinical and biochemical markers for cardiovascular damage risk were measured, and their role as predictors of all-cause and cardiovascular mortality was analysed. RESULTS: NT-proBNP level, ECFv/TBW and other cardiovascular damage risk factors were not evenly distributed among the different dialysis modalities. NT proBNP levels and ECFv/TBW were correlated with several inflammation, malnutrition and myocardial damage markers. Multivariate analysis showed that NT proBNP levels and ECFv/TBW were predictors of both all-cause and cardiovascular mortality, independently of dialysis modality and the presence of other known clinical and biochemical risk factors. CONCLUSIONS: NT-proBNP is a reliable predictor of death risk independently of the effect of dialysis modality on fluid volume control, and the presence of other clinical and biochemical markers recognized as risk factors for all-cause and cardiovascular mortality. NT-pro-BNP is a good predictor of mortality independently of fluid volume overload and dialysis modality. PMID- 19679560 TI - The eye--a window on the kidney. PMID- 19679558 TI - Comparison of methods for estimating glomerular filtration rate in critically ill patients with acute kidney injury. AB - BACKGROUND: In critically ill patients with acute kidney injury, estimates of kidney function are used to modify drug dosing, adjust nutritional therapy and provide dialytic support. However, estimating glomerular filtration rate is challenging due to fluctuations in kidney function, creatinine production and fluid balance. We hypothesized that commonly used glomerular filtration rate prediction equations overestimate kidney function in patients with acute kidney injury and that improved estimates could be obtained by methods incorporating changes in creatinine generation and fluid balance. METHODS: We analysed data from a multicentre observational study of acute kidney injury in critically ill patients. We identified 12 non-dialysed, non-oliguric patients with consecutive increases in creatinine for at least 3 and up to 7 days who had measurements of urinary creatinine clearance. Glomerular filtration rate was estimated by Cockcroft-Gault, Modification of Diet in Renal Disease, Jelliffe equation and Jelliffe equation with creatinine adjusted for fluid balance (Modified Jelliffe) and compared to measured urinary creatinine clearance. RESULTS: Glomerular filtration rate estimated by Jelliffe and Modification of Diet in Renal Disease equation correlated best with urinary creatinine clearances. Estimated glomerular filtration rate by Cockcroft-Gault, Modification of Diet in Renal Disease and Jelliffe overestimated urinary creatinine clearance was 80%, 33%, 10%, respectively, and Modified Jelliffe underestimated GFR by 2%. CONCLUSION: In patients with acute kidney injury, glomerular filtration rate estimating equations can be improved by incorporating data on creatinine generation and fluid balance. A better assessment of glomerular filtration rate in acute kidney injury could improve evaluation and management and guide interventions. PMID- 19679561 TI - RPGRIP1 is essential for normal rod photoreceptor outer segment elaboration and morphogenesis. AB - The function of the retinitis pigmentosa GTPase regulator interacting protein 1 (RPGRIP1) gene is currently not known. However, mutations within the gene lead to Leber Congenital Amaurosis and autosomal recessive retinitis pigmentosa in human patients. In a previously described knockout mouse model of the long splice variant of Rpgrip1, herein referred to as Rpgrip1(tm1Tili) mice, mislocalization of key outer segment proteins and dysmorphogenesis of outer segment discs preceded subsequent photoreceptor degeneration. In this report, we describe a new mouse model carrying a splice acceptor site mutation in Rpgrip1, herein referred to as Rpgrip1(nmf247) that is phenotypically distinct from Rpgrip1(tm1Tili) mice. Photoreceptor degeneration in homozygous Rpgrip1(nmf247) mice is earlier in onset and more severe when compared with Rpgrip1(tm1Tili) mice. Also, ultrastructural studies reveal that whereas Rpgrip1(nmf247) mutants have a normal structure and number of connecting cilia, unlike Rpgrip1(tm1Tili) mice, they do not elaborate rod outer segments (OS). Therefore, in addition to its role in OS disc morphogenesis, RPGRIP1 is essential for rod OS formation. Our study indicates the absence of multiple Rpgrip1 isoforms in Rpgrip1(nmf247) mice, suggesting different isoforms may play different roles in photoreceptors and underscores the importance of considering splice variants when generating targeted null mutations. PMID- 19679562 TI - Parkin selectively alters the intrinsic threshold for mitochondrial cytochrome c release. AB - Autosomal-recessive mutations in the Parkin gene are the second most common cause of familial Parkinson's disease (PD). Parkin deficiency leads to the premature demise of the catecholaminergic neurons of the ventral midbrain in familial PD. Thus, a better understanding of parkin function may elucidate molecular aspects of their selective vulnerability in idiopathic PD. Numerous lines of evidence suggest a mitochondrial function for parkin and a protective effect of ectopic parkin expression. Since mitochondria play a critical role in cell survival/cell death through regulated cytochrome c release and control of apoptosis, we sought direct evidence of parkin function in this pathway. Mitochondria were isolated from cells expressing either excess levels of human parkin or shRNA directed against endogenous parkin and then treated with peptides corresponding to the active Bcl-2 homology 3 (BH3) domains of pro-apoptotic proteins and the threshold for cytochrome c release was analyzed. Data obtained from both rodent and human neuroblastoma cell lines showed that the expression levels of parkin were inversely correlated with cytochrome c release. Parkin was found associated with isolated mitochondria, but its binding per se was not sufficient to inhibit cytochrome c release. In addition, pathogenic parkin mutants failed to influence cytochrome c release. Furthermore, PINK1 expression had no effect on cytochrome c release, suggesting a divergent function for this autosomal recessive PD-linked gene. In summary, these data demonstrate a specific autonomous effect of parkin on mitochondrial mechanisms governing cytochrome c release and apoptosis, which may be relevant to the selective vulnerability of certain neuronal populations in PD. PMID- 19679563 TI - Pluripotency can be rapidly and efficiently induced in human amniotic fluid derived cells. AB - Direct reprogramming of human somatic cells into pluripotency has broad implications in generating patient-specific induced pluripotent stem (iPS) cells for disease modeling and cellular replacement therapies. However, the low efficiency and safety issues associated with generation of human iPS cells have limited their usage in clinical settings. Cell types can significantly influence reprogramming efficiency and kinetics. To date, human iPS cells have been obtained only from a few cell types. Here, we report for the first time rapid and efficient generation of iPS cells from human amniotic fluid-derived cells (hAFDCs) via ectopic expression of four human factors: OCT4/SOX2/KLF4/C-MYC. Significantly, typical single iPS cell colonies can be picked up 6 days after viral infection with high efficiency. Eight iPS cell lines have been derived. They can be continuously propagated in vitro and express pluripotency markers such as AKP, OCT4, SOX2, SSEA4, TRA-1-60 and TRA-1-81, maintaining the normal karyotype. Transgenes are completely inactivated and the endogenous OCT4 promoter is adequately demethylated in the established iPS cell lines. Moreover, various cells and tissues from all three germ layers are found in embryoid bodies and teratomas, respectively. In addition, microarray analysis demonstrates a high correlation coefficient between hAFDC-iPS cells and human embryonic stem cells, but a low correlation coefficient between hAFDCs and hAFDC-iPS cells. Taken together, these data identify an ideal human somatic cell resource for rapid and efficient generation of iPS cells, allowing us to establish human iPS cells using more advanced approaches and possibly to establish disease- or patient-specific iPS cells. PMID- 19679564 TI - Stra13 regulates oxidative stress mediated skeletal muscle degeneration. AB - Duchenne Muscular Dystrophy (DMD), caused by loss of dystrophin is characterized by progressive muscle cell necrosis. However, the mechanisms leading to muscle degeneration in DMD are poorly understood. Here, we demonstrate that Stra13 protects muscle cells from oxidative damage, and its absence leads to muscle necrosis in response to injury in Stra13-deficient mice. Interestingly, Stra13-/- mutants express elevated levels of TNFalpha, reduced levels of heme-oxygenase-1, and display apparent signs of oxidative stress prior to muscle death. Moreover, Stra13-/- muscle cells exhibit an increased sensitivity to pro-oxidants, and conversely, Stra13 overexpression provides resistance to oxidative damage. Consistently, treatment with anti-oxidant N-acetylcysteine ameliorates muscle necrosis in Stra13-/- mice. We also demonstrate that Stra13 expression is elevated in muscles from dystrophin-deficient (mdx) mice, and mdx/Stra13-/- double mutants exhibit an early onset of muscle degeneration. Our studies underscore the importance of oxidative stress-mediated muscle degeneration in muscular dystrophy, and reveal the contribution of Stra13 in maintenance of muscle integrity. PMID- 19679565 TI - The DNA methylome of pediatric acute lymphoblastic leukemia. AB - Acute lymphoblastic leukemia (ALL) is the most common childhood malignancy, with high hyperdiploidy [51-67 chromosomes] and the t(12;21)(p13;q22) [ETV6/RUNX1 fusion] representing the most frequent abnormalities. Although these arise in utero, there is long latency before overt ALL, showing that additional changes are needed. Gene dysregulation through hypermethylation may be such an event; however, this has not previously been investigated in a detailed fashion. We performed genome-wide methylation profiling using bacterial artificial chromosome arrays and promoter-specific analyses of high hyperdiploid and ETV6/RUNX1 positive ALLs. In addition, global gene expression analyses were performed to identify associated expression patterns. Unsupervised cluster and principal component analyses of the chromosome-wide methylome profiles could successfully subgroup the two genetic ALL types. Analysis of all currently known promoter specific CpG islands demonstrated that several B-cell- and neoplasia-associated genes were hypermethylated and underexpressed, indicating that aberrant methylation plays a significant leukemogenic role. Interestingly, methylation hotspots were associated with chromosome bands predicted to harbor imprinted genes and the tri-/tetrasomic chromosomes in the high hyperdiploid ALLs were less methylated than their disomic counterparts. Decreased methylation of gained chromosomes is a previously unknown phenomenon that may have ramifications not only for the pathogenesis of high hyperdiploid ALL but also for other disorders with acquired or constitutional numerical chromosome anomalies. PMID- 19679571 TI - Corneal tattooing (keratopigmentation) with new mineral micronised pigments to restore cosmetic appearance in severely impaired eyes. AB - AIM: To investigate keratopigmentation (KTP) with new mineral micronised pigments as a surgical alternative to improve cosmetic appearance in severely impaired eyes. METHODS: 40 eyes underwent KTP alternatively to invasive cosmetic reconstructive surgery. Corneal staining with mineral micronised pigments was performed using an intralamellar or superficial technique. RESULTS: One year postoperatively, all but two patients (95%) were satisfied. Pigmented eyes were improving patient's appearance. Eight cases needed a second KTP. Two patients with preoperative corneal oedema did not obtain an adequate cosmetic appearance due to progressive pigment clearance observed from 6 months postoperatively. Three eyes with traumatic aniridia observed good cosmetic outcome and a significant reduction in glare. CONCLUSIONS: KTP achieves good cosmetic results and is associated with high patient satisfaction, avoiding extensive and mutilating reconstructive surgery. PMID- 19679572 TI - Optical coherence tomography: new (near-infrared) light on stent implantation? PMID- 19679566 TI - Depletion of zebrafish Tcap leads to muscular dystrophy via disrupting sarcomere membrane interaction, not sarcomere assembly. AB - Tcap/telethonin encodes a Z-disc protein that plays important roles in sarcomere assembly, sarcomere-membrane interaction and stretch sensing. It remains unclear why mutations in Tcap lead to limb-girdle muscular dystrophy 2G (LGMD2G) in human patients. Here, we cloned tcap in zebrafish and conducted genetic studies. We show that tcap is functionally conserved, as the Tcap protein appears in the sarcomeric Z-disc and reduction of Tcap resulted in muscular dystrophy-like phenotypes including deformed muscle structure and impaired swimming ability. However, the observations that Tcap integrates into the sarcomere at a stage after the Z-disc becomes periodic, and that the sarcomere remains intact in tcap morphants, suggest that defective sarcomere assembly does not contribute to this particular type of muscular dystrophy. Instead, a defective interaction between the sarcomere and plasma membrane was detected, which was further underscored by the disrupted development of the T-tubule system. Pertinent to a potential function in stretch sensor signaling, zebrafish tcap exhibits a variable expression pattern during somitogenesis. The variable expression is inducible by stretch force, and the expression level of Tcap is negatively regulated by integrin-link kinase (ILK), a protein kinase that is involved in stretch sensing signaling. Together, our genetic studies of tcap in zebrafish suggested that pathogenesis in LGMD2G is due to a disruption of sarcomere-T-tubular interaction, but not of sarcomere assembly per se. In addition, our data prompted a novel hypothesis that predicts that the transcription level of Tcap can be regulated by the stretch force to ensure proper sarcomere-membrane interaction in striated muscles. PMID- 19679573 TI - Comparing childhood leukaemia treatment before and after the introduction of a parental education programme in Indonesia. AB - SETTING: Previously, treatment and the results of treatment for childhood acute lymphoblastic leukaemia (ALL) in Indonesia differed significantly between poor and prosperous patients. Poor patients received less individual attention from oncologists and access to parental education and donated chemotherapy was lacking. INTERVENTION: A structured parental education programme for both poor and prosperous parents was introduced in January 2004 to improve access to parental education and donated chemotherapy. The programme consisted of a video presentation, an information booklet, DVD, audiocassette, a statement-of understanding for donated chemotherapy, and a complaints procedure. Informed consent was also sought. OBJECTIVE: Our study compared childhood ALL treatment outcome before and after the introduction of the parental education programme. DESIGN: The medical records of 283 children with ALL diagnosed before (1997-2002; n = 164) and after (2004-2006; n = 119) the introduction of the education programme were reviewed. Data on treatment results and parental socioeconomic status were collected. RESULTS: After the introduction of the education programme, treatment refusal decreased (from 14% to 2%) and event-free survival increased (from 13% to 29%) significantly among poor patients. Treatment dropout increased (from 0% to 13%) significantly among prosperous patients. Overall, toxic death (from 23% to 36%) increased significantly, but there was no significant difference in event-free survival. CONCLUSIONS: After introduction of the programme, treatment refusal decreased and event-free survival increased significantly among poor families. However, improved knowledge, skills and communication are still required to combat the high rates of toxic death and treatment dropout. Treatment intensity should be accompanied by improved supportive care. PMID- 19679574 TI - Difficulties in selecting an appropriate neonatal thyroid stimulating hormone (TSH) screening threshold. AB - BACKGROUND: The UK Newborn Screening Programme Centre recommends that a blood spot thyroid stimulating hormone (TSH) cut-off of 10 mU/l is used to detect congenital hypothyroidism (CHT). As the value used varies from 5 to 10 mU/l, we examined the implications of altering this threshold. METHODS: Our regional blood spot TSH cut-off is 6 mU/l. Positive or suspected cases were defined as a TSH >6 mU/l throughout the study period (1 April 2005 to 1 March 2007). All term infants (>35 weeks) whose first TSH was 6-20 mU/l had a second TSH measured. The biochemical details of infants with a TSH between 6.1 and 10.0 mU/l and then >6 mU/l on second sampling were sent to paediatric endocrinologists to determine approaches to management. RESULTS: 148 of 65 446 infants (0.23%) had a first blood spot TSH >6.0 mU/l. 120 were term infants with 67 of these (0.1% of all infants tested) having a TSH between 6.1 and 10.0 mU/l and 53 a TSH >10.0 mU/l. Of the 67 term infants with a TSH between 6.1 and 10.0 mU/l on initial testing, four continued to have a TSH >6 mU/l. One with a TSH >10 mU/l and one infant with a TSH <10 mU/l on the second blood spot have been diagnosed with CHT. The survey of endocrinologists highlighted significant differences in practice. CONCLUSIONS: A reduced threshold of 6 mU/l will increase the number of false positive term infants by 126%, but abnormalities of thyroid function requiring treatment will be detected. We suspect that the additional expense involved in setting a lower threshold is justified. PMID- 19679575 TI - Clinical effectiveness of customised sport shoe orthoses for overuse injuries in runners: a randomised controlled study. AB - BACKGROUND AND OBJECTIVES: Treatment of chronic running-related overuse injuries by orthopaedic shoe orthoses is very common but not evidence-based to date. HYPOTHESIS: Polyurethane foam orthoses adapted to a participant's barefoot plantar pressure distribution are an effective treatment option for chronic overuse injuries in runners. DESIGN: Prospective, randomised, controlled clinical trial. INTERVENTION: 51 patients with running injuries were treated with custom made, semirigid running shoe orthoses for 8 weeks. 48 served as a randomised control group that continued regular training activity without any treatment. MAIN OUTCOME MEASURES: Evaluation was made by the validated pain questionnaire Subjective Pain Experience Scale, the pain disability index and a comfort index in the orthoses group (ICI). RESULTS: There were statistically significant differences between the orthoses and control groups at 8 weeks for the pain disability index (mean difference 3.2; 95% CI 0.9 to 5.5) and the Subjective Pain Experience Scale (6.6; 2.6 to 10.6). The patients with orthoses reported a rising wearing comfort (pre-treatment ICI 69/100; post-treatment ICI 83/100) that was most pronounced in the first 4 weeks (ICI 80.4/100). CONCLUSION: Customised polyurethane running shoe orthoses are an effective conservative therapy strategy for chronic running injuries with high comfort and acceptance of injured runners. PMID- 19679576 TI - Teleoanticipation in all-out short-duration cycling. AB - OBJECTIVE: to investigate the effect of all-out cycling test duration on indices of power, anaerobic lactic energy metabolism, perceived exertion and mood. METHODS: nine physically active men undertook four all-out cycling tests of 5, 15, 30 or 45 s from seated stationary start on an ergometer fit with power cranks. The participants completed a Profile of Mood States questionnaire before each test and indicated perceived exertion immediately post-test (Borg 6-20 scale). Indices of anaerobic lactic metabolism were determined from blood lactate concentrations. RESULTS: pacing strategy was apparent in the 45-s tests with lower peak (p<0.01) and mean power in the initial 10 s compared to the 5- and 15 s tests (p<0.05). The first 15 s of the 30- and 45-s tests revealed lower fatigue indices compared to the 15-s tests (p<0.05) indicating some pacing in the 30-s tests. Perceived exertion increased with duration, with no difference between the 15- and 30-s tests (p>0.05). Extravascal lactate generation (reflecting exercising muscle lactate production) explained 59% of the variance in perceived exertion. There was no effect of knowledge of test duration on mood states or total mood disturbance (p>0.05). CONCLUSIONS: an all-out pacing strategy was apparent for at least up to 15 s, with indicators of dampened power in both 30 and 45 s sprints. Reduced power at the start of all-out long-duration sprints support a central control of at least initial pacing strategy. PMID- 19679577 TI - Establishing maximal oxygen uptake in young people during a ramp cycle test to exhaustion. AB - OBJECTIVES: This study tested the hypotheses that (1) secondary criteria (respiratory exchange ratio (RER), heart rate, blood [lactate]) traditionally used to verify the determination of maximum oxygen uptake (VO2(max)) in children can result in the acceptance of a 'submaximal' VO2(max) or falsely reject a 'true' VO(2max) and (2) the VO2(peak) recorded during a ramp test in children is comparable to the VO2(peak) achieved during supramaximal testing. METHODS: Thirteen children (9-10 years) completed a ramp cycle test to exhaustion to determine their VO2(peak). After 15 min of recovery, the participants performed a supramaximal cycle test to exhaustion at 105% of their ramp test peak power. RESULTS: Compared with the VO2(peak) during the ramp test, a significantly lower VO2 was recorded at a RER of 1.00 (1.293 litre/min (SD 0.265) vs 1.681 litre/min (SD 0.295), p < 0.001, n = 12), at a heart rate of 195 beats/min (1.556 litre/min (SD 0.265) vs 1.721 litre/min (SD 0.318), p < 0.001, n = 10) and at 85% of age predicted maximum (1.345 litre/min (SD 0.228) vs 1.690 litre/min (SD 0.284), p < 0.001, n = 13). Supramaximal testing yielded a VO2(peak) that was not significantly different from the ramp test (1.615 litre/min (SD 0.307) vs 1.690 litre/min (SD 0.284), p = 0.090, respectively). CONCLUSIONS: The use of secondary criteria to verify a maximal effort in young people during ramp cycling exercise may result in the acceptance of a submaximal VO2(max). As supramaximal testing elicits a VO2(peak) similar to the ramp protocol, thus satisfying the plateau criterion, the use of such tests is recommended as the appropriate method of confirming a 'true' VO2(max) with children. PMID- 19679578 TI - The effect of mindfulness meditation on cough reflex sensitivity. AB - BACKGROUND: Chronic cough is common, and medical treatment can be ineffective. Mindfulness is a psychological intervention that aims to teach moment-to-moment non-judgemental awareness of thoughts, feelings and sensations. METHOD: 30 healthy subjects and 30 patients with chronic cough were studied in two sequential trials. For both studies, cough reflex sensitivity to citric acid (C5) was measured on two occasions, with urge to cough rated following each inhalation; between challenges subjects were randomised to (1) no intervention, (2) mindfulness or (3) no intervention but modified cough challenge (subjects suppress coughing). For the healthy volunteers, measures were 1 h apart and mindfulness was practised for 15 min. For the patients with chronic cough measures were 1 week apart and mindfulness was practised daily for 30 min. RESULTS: In healthy volunteers, median change (interquartile range (IQR)) in cough reflex sensitivity (logC5) for no intervention, mindfulness and suppression was +1.0 (0.0 to +1.3), +2.0 (+1.0 to +3.0) and +3.0 (+2.8 to +3.0) doubling concentrations (p = 0.003); there were significant reductions for both mindfulness (p = 0.043) and suppression (p = 0.002) over no intervention. In patients with cough, median change (IQR) in logC5 for no intervention, mindfulness training and voluntary suppression was 0.0 (-1.0 to +1.0), +1.0 (-0.3 to +1.0) and +1.0 (+1.0 to +2.0) doubling concentrations (p = 0.046); there was a significant reduction for suppression (p = 0.02) but not mindfulness (p = 0.35). Urge to cough did not change after mindfulness compared with control in either healthy subjects (p = 0.33) or those with chronic cough (p = 0.47). CONCLUSION: Compared with control, mindfulness decreased cough reflex sensitivity in healthy volunteers, but did not alter cough threshold in patients with chronic cough. Both groups were able to suppress cough responses to citric acid inhalation. PMID- 19679579 TI - Obstructive sleep apnoea is associated with diabetes in sleepy subjects. AB - BACKGROUND: Although obstructive sleep apnoea (OSA) has been linked to insulin resistance and glucose intolerance, it is unclear whether there is an independent association between OSA and diabetes mellitus (DM) and whether all patients with OSA are at risk. The objective of this study was to determine the association between OSA and DM in a large cohort of patients referred for sleep diagnostic testing. METHODS: A cross-sectional analysis of participants in a clinic-based study was conducted between July 2005 and August 2007. DM was defined by self report and concurrent use of diabetic medications (oral hypoglycaemics and/or insulin). Sensitivity analysis was performed using a validated administrative definition of diabetes. OSA was defined by the respiratory disturbance index (RDI) using polysomnography or ambulatory monitoring. Severe OSA was defined as an RDI > or = 30/h. Subjective sleepiness was defined as an Epworth Sleepiness Scale score > or = 10. RESULTS: Complete data were available for 2149 patients. The prevalence of DM increased with increasing OSA severity (p<0.001). Severe OSA was associated with DM following adjustment for patient demographics, weight and neck circumference (odds ratio (OR) 2.18; 95% CI 1.22 to 3.89; p<0.01). Following a stratified analysis, this relationship was observed exclusively in sleepy patients (OR 2.59 (95% CI 1.35 to 4.97) vs 1.16 (95% CI 0.31 to 4.37) in non sleepy patients). CONCLUSIONS: Severe OSA is independently associated with DM in patients who report excessive sleepiness. Future studies investigating the impact of OSA treatment on DM may wish to focus on this patient population. PMID- 19679580 TI - Cardiovascular reflex responses to temporal reduction in arterial pressure during dexmedetomidine infusion: a double-blind, randomized, and placebo-controlled study. AB - BACKGROUND: The low and moderate doses of dexmedetomidine reduce arterial pressure and heart rate (HR), suggesting attenuation of sympathetic activity and dominance of cardiac-vagal activity. These autonomic responses under dexmedetomidine sedation may attenuate cardiovascular reflex responses to temporal reduction in arterial pressure, inducing a severe hypotension. We therefore investigated the effects of dexmedetomidine on cardiovascular reflex responses to temporal reduction in arterial pressure induced by the thigh cuff method. METHODS: Twelve healthy men received placebo, low-dose (loading 3 microg kg(-1) h(-1) for 10 min; maintenance 0.2 microg kg(-1) h(-1) for 60 min), and moderate-dose (loading 6 microg kg(-1) h(-1) for 10 min; maintenance 0.4 microg kg(-1) h(-1) for 60 min) dexmedetomidine infusions in a randomized, double-blind, crossover study. After 70 min of drug infusion, systolic arterial pressure (SAP) and HR responses after thigh cuff deflation were evaluated as indices of cardiovascular reflex. RESULTS: Reduction in SAP (Delta SAP) [placebo 8 (4), low 12 (4), moderate 19 (5) mm Hg] after thigh cuff deflation was significantly greater in dexmedetomidine than placebo infusions, in a dose-dependent manner. The change in HR (Delta HR), Delta HR/Delta SAP, and the percentage restoration of SAP were lower with dexmedetomidine compared with placebo. CONCLUSIONS: The present results indicated that dexmedetomidine weakens arterial pressure preservation and HR responses after thigh cuff deflation, suggesting attenuated cardiovascular reflexes. Therefore, it must be cautioned that dexmedetomidine can lead to further and sustained reduction in arterial pressure during transient hypotension induced by postural changes, haemorrhage, and/or other stresses. PMID- 19679582 TI - Does strict glucose control improve outcome? PMID- 19679583 TI - Antiseptic solutions for central neuraxial blockade: which concentration of chlorhexidine in alcohol should we use? PMID- 19679584 TI - Management of hypotension in obstetric spinal anaesthesia. PMID- 19679585 TI - Securing tracheal tubes in facial burns. PMID- 19679586 TI - Use of Luer connection syringes for spinal anaesthesia. PMID- 19679587 TI - Anatomical causes of failed spinal anaesthesia may be commoner than thought. PMID- 19679588 TI - Anaesthesia for a patient with Isaac's syndrome and myasthenia gravis. PMID- 19679589 TI - Apparent blind spot with the GlideScope video laryngoscope. PMID- 19679590 TI - Dabigatran etexilate: future directions in anticoagulant treatment. AB - Dabigatran etexilate is a novel, oral reversible direct thrombin inhibitor in the clinical development for the treatment and prevention of thromboembolic diseases. Clinical data indicate that dabigatran etexilate has immediate onset of effect, no need for monitoring, predictable and consistent pharmacokinetics and pharmacodynamics-all features that differentiate it from oral vitamin K antagonists (VKAs). Completed phase III studies demonstrated a comparable efficacy and safety profile to enoxaparin in the prevention of venous thromboembolism (VTE) after orthopedic surgery. Ongoing phase III trials are now evaluating the long-term use of dabigatran etexilate for the treatment and secondary prevention of VTE and for prevention of stroke in patients with atrial fibrillation, as a replacement for VKAs. With an immediate, reliable, and predictable anticoagulant effect without the need for coagulation monitoring and the lack of long-term safety concerns, dabigatran etexilate may be a prospective candidate that offers additional benefit over VKAs and parenteral anticoagulants in these settings. PMID- 19679591 TI - The effects of hypothyroidism on the mechanical properties and histomorphological structure of the thoracic aorta. AB - This experimental study investigates the effects of hypothyroidism on the descending thoracic aorta. Hypothyroidism was induced in 20 male Wistar rats by administering 0.05% of 6-n propyl 2-thiouracil (PTU) in their drinking water for 8 weeks. Euthyroid rats were used as controls. Animals were sacrificed and longitudinal strips of the descending aorta were subjected to various preselected levels of stress in a uniaxial tensile-testing device. Analysis of stress-strain, elastic modulus-strain curves disclosed significant differences between groups, indicative of stiffer aortas in hypothyroid animals at the upper physiologic and higher levels of pressure. Remodeling of the aortic wall of hypothyroid animals revealed significant histological changes. The thoracic aorta of hypothyroid rats compared with that of euthyroid ones became stiffer at high strains, including the upper physiologic range, loosing part of its distensibility. Hypothyroidism was also associated with diameter enlargement and substantial lengthening of the aorta. PMID- 19679592 TI - Demographic adjustments for the Spanish version of the WAIS-III. AB - The Spanish version of the third edition of the Wechsler Adult Intelligence Scale (WAIS-III) by TEA Ediciones is an excellent addition to available instruments for Spanish speakers. The Spanish norms function similarly to US norms for individuals aged 16-35. The norms become increasingly different for individuals 35 and older, seemingly because of the lower levels of formal education of the older Spanish cohorts. Using data from a random half of the Spanish sample, the authors developed regression equations to adjust the scaled scores for individuals with a low level of education. The adjustment is made to the level that would have been expected if the individual had 12 years of education, the median level of education of the US norms. The article includes the methodology and values necessary to make the adjustments. The scaled scores were then adjusted for individuals on the second random half of the Spanish sample and compared to the United States norms. The results showed the adjustments succeed in bringing the Spanish norms closer to the US norms. PMID- 19679593 TI - The Indiana faces in places test: preliminary findings on a new visuospatial memory test in patients with mild cognitive impairment. AB - Memory assessment is an important component of a neuropsychological evaluation, but far fewer visual than verbal memory instruments are available. We examined the preliminary psychometric properties and clinical utility of a novel, motor free paper and pencil visuospatial memory test, the Indiana faces in places test (IFIPT). The IFIPT and general neuropsychological performance were assessed in 36 adults with amnestic mild cognitive impairment (aMCI) and 113 older adults with no cognitive impairment at baseline, 1 week, and 1 year. The IFIPT is a visual memory test with 10 faces paired with spatial locations (three learning trials and non-cued delayed recall). Results showed that MCI participants scored lower than controls on several variables, most notably total learning (p < .001 at all three time points), delayed recall (baseline p = .03, 1 week p < .001, 1 year p < .001), and false-positive errors (range p = .03 to <0.001). The IFIPT showed similar test-retest reliability at 1-week and 1-year follow-up to other neuropsychological tests (r = 0.71-0.84 for MCI and 0.53-0.72 for controls). Diagnostic accuracy was modest for this sample (areas under the receiver operating characteristic curve between 0.64 and 0.66). Preliminary psychometric analyses support further study of the IFIPT. The measure showed evidence of clinical utility by demonstrating group differences between this sample of healthy adults and those with MCI. PMID- 19679595 TI - FV100 as a new approach for the possible treatment of varicella-zoster virus infection. AB - FV100 is a promising new agent with extreme potency and specificity for varicella zoster virus (VZV). It is the valyl ester pro-drug of Cf1743, the lead clinical candidate among the highly lipophilic bicyclic nucleoside analogue (BCNA) family discovered in Cardiff/Leuven. Cf1743 is unique amongst antivirals in terms of its structure and lipophilicity. It is exquisitely potent and selective for human VZV. FV100 has recently entered a randomized, controlled Phase II clinical trial for the treatment of shingles, sponsored by Inhibitex. PMID- 19679594 TI - Development of set-shifting ability from late childhood through early adulthood. AB - This cross-sectional study examined the development of set-shifting ability from childhood into early adulthood. Six hundred and forty-nine participants (aged 8 30) were assessed on the verbal fluency, design fluency, trail making, color-word interference, and card sorting subtests of the Delis-Kaplan Executive Function System (D-KEFS). Multiple regression analyses revealed modest effects of age and gender on set-shifting tasks, after controlling for IQ and component skills. The current study provides evidence for generally increased performance of set shifting abilities through adolescence. Women overall had statistically better performance than men on all executive functioning tasks. There were significant age by gender interactions suggesting differential age-related improvements between men and women. On color-word interference and verbal fluency switching tasks, men tended to show larger improvements than women, whereas on a design fluency switching task, women showed larger improvements than men. PMID- 19679596 TI - Organic solvent tolerance and fluoroquinolone resistance in Klebsiella pneumoniae clinical isolates. PMID- 19679598 TI - Medication errors: how reliable are the severity ratings reported to the national reporting and learning system? AB - OBJECTIVE: To examine: (1) the reliability of the severity rating scale used by the National Reporting and Learning System (NRLS) in England and Wales for medication errors; and (2) the likelihood of reporting medication errors among healthcare professionals. SETTING: A 900-bed acute university teaching hospital in the North West of England. PARTICIPANTS: Forty healthcare professionals (10 doctors, 10 nurses, 10 pharmacists and 10 pharmacy technicians). METHODS: Participants were asked to complete a self-administered questionnaire containing nine medication error scenarios on two separate occasions. They were asked to rate the severity of each incident using the NRLS severity rating scale and also the likelihood of reporting the incident via the hospital incident reporting system. The MAIN OUTCOME MEASURES: included comparisons of severity ratings and likelihood of reporting by the four health professional groups. Test-retest reliability of the severity ratings was also examined within and between professional groups. RESULTS: Pharmacists and nurses were significantly more likely to report the errors if they had witnessed them (mean scores 36.3 and 36.2, respectively, compared with 27.9 for doctors, P < 0.001). Nurses and pharmacy technicians assigned higher severity ratings for medication errors (mean scores 23.6 and 25, respectively) than pharmacists or doctors (both 19.4). Both within and between healthcare professional groups, there was wide variation in the assignment of medication error severity ratings. CONCLUSIONS: There are marked differences in the severity ratings for medication errors graded against the NRLS severity criteria between different health professional groups and at different time points rated by the same individuals. PMID- 19679597 TI - Antibacterial activities of a fosfomycin/tobramycin combination: a novel inhaled antibiotic for bronchiectasis. AB - OBJECTIVES: To compare the in vitro and in vivo activities of a 4:1 (w/w) fosfomycin/tobramycin combination (FTI) with those of fosfomycin and tobramycin alone against cystic fibrosis (CF) and non-CF bronchiectasis pathogens. METHODS: Clinical isolates of CF Pseudomonas aeruginosa, Staphylococcus aureus, Haemophilus influenzae, Stenotrophomonas maltophilia, Burkholderia cepacia complex, Escherichia coli and Klebsiellia spp. were evaluated by MIC, MBC, post antibiotic effect (PAE), synergy, time-kill, a rat pneumonia model and spontaneous mutation frequency (SMF). RESULTS: FTI showed high activity against E. coli, H. influenzae, S. aureus and Klebsiella spp. For the S. aureus strains, 75% of which were methicillin resistant (MRSA), FTI had a lower MIC(90) than tobramycin. For P. aeruginosa, FTI had a lower MIC(90) than fosfomycin, but tobramycin was more active than either. Synergy studies showed no antagonism between fosfomycin and tobramycin, and 93% of the isolates demonstrated no interaction. FTI was rapidly bactericidal and exhibited concentration-dependent killing in time-kill studies. In the rat pneumonia model, FTI and tobramycin demonstrated bactericidal killing of P. aeruginosa; both were more active than fosfomycin alone. The SMF for S. aureus resistance to FTI was 2-4 log(10) lower than that for tobramycin and 2-7 log(10) lower than that for fosfomycin. For P. aeruginosa, the FTI SMF was 2-3 log(10) lower than that for fosfomycin and 1-2 log(10) lower than that for tobramycin. CONCLUSIONS: FTI is a broad-spectrum antibiotic combination with high activity in vitro and in vivo. These data suggest FTI could be a potential treatment for respiratory infections caused by gram-positive and gram-negative aerobic bacteria. PMID- 19679599 TI - Myeloid RelA regulates pulmonary host defense networks. AB - The pulmonary innate immune system in the respiratory tract eliminates inhaled pathogens. Several cell types contribute to host defense within a complex network. The aim of this study was to evaluate the role of macrophages during pneumonia and in the regulation of the epithelial response to microorganisms. We performed lung infection models in mice lacking myeloid RelA/p65. To study the mechanistic relationships between individual cell types, we applied co-culture models composed of airway epithelial cells (AECs) and macrophages. Mice lacking myeloid RelA/p65 showed significantly decreased bacterial clearance, cytokine expression and neutrophil influx. In addition, the induction of epithelial keratinocyte chemoattractant expression was blunted in these animals. In vitro, AECs were largely insensitive to ligands of Toll-like receptor (TLR)2 or TLR5. Exposure to secretory products of macrophages results in an increased release of pro-inflammatory cytokines and augmented antimicrobial activity. This was associated with increased expression of TLR genes and surface expression of the proteins. Experiments with blocking antibodies showed that the effect of macrophages depends on secreted mediators, including tumour necrosis factor alpha. In conclusion, the present data show that myeloid RelA is critical for pulmonary host defense. One important mechanism is that macrophages induce the sensitivity of AEC's to microbial patterns. PMID- 19679601 TI - Community-acquired pneumonia through Enterobacteriaceae and Pseudomonas aeruginosa: Diagnosis, incidence and predictors. AB - The aim of the present study was to determine the relevance of the presence of Enterobacteriaceae (EB) and Pseudomonas aeruginosa (PA) in patients with community-acquired pneumonia (CAP) and how the true incidence of these pathogens can be assessed. Based on prospective data from 5,130 patients with CAP included in the German Competence Network for Community-Acquired Pneumonia (CAPNETZ), the incidence, clinical characteristics, outcome and predictors of patients with CAP due to EB and PA were studied applying strict case definitions. The incidence of EB was 67 (1.3%) out of 5,130, including 27 patients with bacteraemia. PA was found in 22 (0.4%) out of 5,130 patients. These microorganisms were judged to be indeterminate pathogens in an additional 172 and 27 isolates, respectively. Patients with indeterminate pathogens differed considerably from those with definite isolates in terms of clinical presentation, comorbidity, pneumonia severity and outcome. Independent risk factors for EB included cardiac and cerebrovascular disease, and for PA chronic respiratory disease and enteral tube feeding. The 30-day mortality was significantly higher in patients with definite pathogens. In the present large population, the incidence of CAP due to EB/ PA was low. The risk of the presence of these pathogens can be assessed using several predictors, which may identify those patients in need of an extended diagnostic work-up and initial antimicrobial treatment. PMID- 19679600 TI - Quality of life and dyspnoea in patients treated with bosentan for idiopathic pulmonary fibrosis (BUILD-1). AB - No therapy is known to improve health-related quality of life (HRQoL) or dyspnoea in patients with idiopathic pulmonary fibrosis. The present study investigated longitudinal changes in HRQoL and dyspnoea and explored the effects of bosentan on these end-points during the Bosentan Use in Interstitial Lung Disease (BUILD) 1 trial. In total, 154 subjects received oral bosentan (n = 71) or placebo (n = 83). Changes in HRQoL and dyspnoea from baseline to month (M) 6 and up to M12 were measured using the St George's Respiratory Questionnaire (SGRQ), 36-item short-form health survey (SF-36), Transition Dyspnoea Index and Borg dyspnoea index. Overall, minimal changes occurred in measures of HRQoL and dyspnoea among placebo-treated subjects during the study. The effects of bosentan treatment on HRQoL and dyspnoea in the all-treated population were minimal. However, in the subset of subjects who had undergone surgical lung biopsy for diagnosis of idiopathic pulmonary fibrosis, treatment effects were observed up to M12 in the impact domain of the SGRQ and the physical functioning, general health and role emotional domains of the SF-36. HRQoL and dyspnoea changed minimally during the course of the present study. Observations from exploratory analyses suggested benefits of bosentan on HRQoL among patients who had undergone surgical lung biopsy for diagnosis, and they merit further investigation. PMID- 19679602 TI - Effects of cannabis on lung function: a population-based cohort study. AB - The effects of cannabis on lung function remain unclear and may be different from those of tobacco. We compared the associations between use of these substances and lung function in a population-based cohort (n = 1,037). Cannabis and tobacco use were reported at ages 18, 21, 26 and 32 yrs. Spirometry, plethysmography and carbon monoxide transfer factor were measured at 32 yrs. Associations between lung function and exposure to each substance were adjusted for exposure to the other substance. Cumulative cannabis use was associated with higher forced vital capacity, total lung capacity, functional residual capacity and residual volume. Cannabis was also associated with higher airway resistance but not with forced expiratory volume in 1 s, forced expiratory ratio or transfer factor. These findings were similar among those who did not smoke tobacco. In contrast, tobacco use was associated with lower forced expiratory volume in 1 s, lower forced expiratory ratio, lower transfer factor and higher static lung volumes, but not with airway resistance. Cannabis appears to have different effects on lung function from those of tobacco. Cannabis use was associated with higher lung volumes, suggesting hyperinflation and increased large-airways resistance, but there was little evidence for airflow obstruction or impairment of gas transfer. PMID- 19679603 TI - Lung function predicts lung cancer risk in smokers: a tool for targeting screening programmes. AB - The relationship between smoking, lung cancer and airflow obstruction is recognised but it is unclear whether the presence of minimal lung function damage constitutes an independent risk factor for the development of lung cancer. In order to identify those individuals at higher risk of lung cancer on the basis of functional impairment, we evaluated baseline pulmonary function tests of 3,806 heavy smokers undergoing annual chest computed tomography screening, and compared the forced expiratory volume in 1 s (FEV(1)) % predicted of 57 lung cancer cases and that of 3,749 subjects without cancer. We obtained odds ratios (ORs) of lung cancer and the corresponding 95% confidence intervals (CIs) using unconditional logistic regression, adjusting for age, sex, study and smoking variables. Compared with subjects with FEV(1) >or=90% pred, the OR of lung cancer was 2.45 (95% CI 1.39-4.33) for subjects with FEV(1) <90% pred and 2.90 (95% CI 1.34-6.27) for subjects with FEV(1) <70% pred. These data show that even a relatively small reduction in FEV(1) % pred is a significant predictor of increased lung cancer risk. Test screening for lung cancer using airflow obstruction with FEV(1) <90% is a strategy worth future consideration. PMID- 19679604 TI - Carbon monoxide pollution is associated with decreased lung function in asthmatic adults. AB - The aim of the present study was to test the effects of exposure to air pollutants on lung function. A panel of 19 adult asthmatics living in Padua (Italy) was followed for five 30-day periods during 2 yrs consecutively (1,492 morning and 1,434 evening measures analysed). Peak expiratory flow (PEF) and forced expiratory volume in 1 s (FEV(1)) were measured using a pocket electronic meter. Daily levels of air pollutants and meteorological variables were collected at outdoor city monitoring sites. Significant inverse associations were observed between morning and evening PEF and carbon monoxide level (p = 0.01-0.03), without clear differences between lags (0-3 days). An increment of 1 mg.m(-3) CO was associated with a PEF variation ranging -2.6- -2.8%. All effect estimates on PEF for CO remained significant and even increased after controlling for particles with a 50% cut-off aerodynamic diameter of 10 microm (PM(10)), nitrogen dioxide and sulphur dioxide in single and multi-pollutant models. A similar trend was observed for FEV(1), but the associations were nonsignificant. A nonsignificant inverse relationship between evening PEF and SO(2) was also detected. PEF and FEV(1) were not related to PM(10) and NO(2) concentrations. The present results indicate that, in this panel of adult asthmatics, the worsening of lung function is associated with exposure to gaseous pollutants and occurs at levels of CO and SO(2) lower than current European standards. PMID- 19679605 TI - Acute inflammatory response secondary to intrapleural administration of two types of talc. AB - Intrapleural instillation of talc has been used in the treatment of recurrent pleural effusions but can, in rare instances, result in respiratory failure. Side effects seem to be related to composition, size and inflammatory power of talc particles. The aim of this study was to evaluate the inflammatory response to intrapleural injection of talc containing small particles (ST) or talc containing particles of mixed size (MT). 100 rabbits received intrapleural talc, 50 with ST (median 6.41 mum) and 50 with MT (median 21.15 mum); the control group was composed of 35 rabbits. Cells, lactate dehydrogenase, C-reactive protein (CRP), interleukin (IL)-8 and vascular endothelial growth factor were evaluated in serum and bronchoalveolar lavage at 6, 24, 48, 72 and 96 h. Lung histology and the presence of talc were also analysed. Statistics were performed using ANOVA and an unpaired t-test. Most of the parameters showed greater levels in the animals injected with talc than in the controls, suggesting a systemic and pulmonary response. Higher serum levels of CRP and IL-8 were observed in the animals injected with ST. Talc particles were observed in both lungs with no differences between groups. Lung cell infiltrate was more evident in the ST group. In conclusion, talc with larger particles should be the preferred choice in clinical practice in order to induce safer pleurodesis. PMID- 19679606 TI - Beta(2)-agonists block tussive responses in guinea pigs via an atypical cAMP dependent pathway. AB - beta(2)-Adrenoceptor agonists are the most effective bronchodilators currently available, and are used for symptom management in asthmatics. However, whether beta(2)-agonists are also antitussive is controversial. Identifying an antitussive role for beta(2)-agonists and dissecting the possible mechanism of action may help to explain the inconsistencies in the clinical literature and lead to the development of novel therapeutic agents. The aim of the present study was to determine whether or not beta(2)-agonists attenuate the tussive response in guinea pig and human models, and, if so, to identify the mechanism(s) involved. Depolarisation of vagal sensory nerves (human and guinea pig) was assessed as an indicator of sensory nerve activity. Cough was measured in a conscious guinea pig model. A beta(2)-agonist, terbutaline, dose-dependently inhibited the cough response to tussive agents in conscious guinea pigs. Terbutaline and another beta(2)-agonist, fenoterol, blocked sensory nerve activation in vitro. Using these mechanistic models, it was established that beta(2)-agonists suppress the tussive response via a nonclassical cyclic adenosine monosphosphate-dependent pathway that involves the activation of protein kinase G and, subsequently, the opening of large-conductance calcium activated potassium channels. In conclusion, beta(2)-adrenoceptor agonists are antitussive, and this property occurs due to a direct inhibition of sensory nerve activation. These findings may help to explain the confusion that exists in the clinical literature, and could be exploited to identify novel therapies for the treatment of cough, which is a significant unmet medical need. PMID- 19679607 TI - Airway proteins involved in bacterial clearance susceptible to cathepsin G proteolysis. AB - Serine proteases released from neutrophils are central to the pathogenesis of cystic fibrosis lung disease and are considered to be obvious therapeutic targets. Neutrophil elastase digests key opsonins present in the lung and disrupts phagocytosis, allowing bacteria to persist despite established pulmonary inflammation. We have found that cathepsin G, an abundant serine protease found in human and murine neutrophils, has other roles in the development of suppurative lung diseases. Murine models of endobronchial inflammation indicate that cathepsin G inhibits airway defences and interferes with the host's ability to clear Pseudomonas aeruginosa from the lung with effects distinct from neutrophil elastase. We hypothesise that differences in bacterial killing are due to defects in innate defences created by proteolysis. Protein profiles of bronchoalveolar lavage of infected wild-type and cathepsin G-deficient mice were compared using two-dimensional polyacrylamide gel electrophoresis and tandem mass spectrometry. Four proteins in bronchoalveolar lavage were cleaved by cathepsin G. Serum amyloid P component leaked into the lung during acute infection and was digested by cathepsin G. Its cleavage products had greater binding to lipopolysaccharide and interfered with phagocytosis. These results indicate that cleaved serum amyloid P component acts as an anti-opsonin and interferes with bacterial clearance from the lung. PMID- 19679608 TI - CARD15/NOD2 polymorphisms are associated with severe pulmonary sarcoidosis. AB - Sarcoidosis and Crohn's disease are heterogeneous systemic diseases characterised by granulomatous inflammation. Caspase recruitment domain (CARD)15 is a major susceptibility gene for Crohn's disease, and specifically for ileal and fibrostenotic subtypes. The C-C chemokine receptor (CCR)5 gene has been associated with both parenchymal pulmonary sarcoidosis and perianal Crohn's disease. This study explored associations between CARD15 polymorphisms, CCR5 haplotype and distinct pulmonary sarcoidosis subtypes. 185 Caucasian sarcoidosis patients were genotyped for CARD15 and CCR5 polymorphisms. The genetic data were compared with 347 healthy controls and were examined for associations with serial pulmonary function tests and chest radiographs. CARD15 genotypes did not differ between the unselected sarcoidosis cohort and controls. However, patients carrying the functional 2104T (702W) polymorphism were more likely to have radiographic stage IV disease at 4-yr follow-up. All patients possessing both CARD15 2104T and CCR5 HHC haplotype had stage IV disease at presentation. Carriage of 2104T was associated with worse forced expiratory volume in 1 s, whereas carriage of the CARD15 1761G (587R) polymorphism was associated with better lung function. For the first time, an association between two CARD15 polymorphisms and specific sarcoidosis phenotypes has been demonstrated, as well as an additive effect of possessing CARD15 2104T and CCR5 HHC haplotype. PMID- 19679609 TI - Leukotriene D4 activates {beta}2-integrin adhesion in human polymorphonuclear leukocytes. AB - We examined the functional role and mechanisms by which activation of cysteinyl leukotriene-1 receptor (cysLT(1)R) regulates beta(2)-integrin adhesion to intercellular adhesion molecule (ICAM)-1 in human polymorphonuclear leukocytes (PMNs) in vitro. Human peripheral blood PMNs and eosinophils were isolated separately from the same mildly atopic donors. Surface expression of cysLT(1)R was identified both in PMNs and in eosinophils by immunofluorescence analysis. Total cysLT(1)R protein was substantially greater in eosinophils than in PMNs as determined by Western blot analysis. However, leukotriene D(4) (LTD(4)) upregulated beta(2)-integrin adhesion of PMNs to ICAM-1 with high efficacy in a time- and concentration-dependent manner. Upregulated beta(2)-integrin adhesion of PMNs was related temporally and quantitatively to phosphorylation of 85-kDa cytosolic group IVa phospholipase A2 (gIVaPLA2). Augmented LTD(4)-induced adhesion was blocked significantly by montelukast, a cysLT(1)R antagonist. Trifluoromethylketone (a gIVaPLA2 inhibitor) blocked beta(2)-integrin adhesion caused by LTD(4) activation, as did anti-CD18 monoclonal antibody directed against beta(2)-integrin on the PMN surface. Our data demonstrate that LTD(4) causes phosphorylation of gIVaPLA2 and upregulation of beta(2)-integrin adhesion to ICAM-1 or ICAM-1 surrogate through cysLT(1)R activation. Activation of gIVaPLA2 is a critical step through which beta(2)-integrin adhesion is upregulated by the cysLT(1)R expressed on the surface membrane of human PMN. PMID- 19679610 TI - Anomalous left coronary artery from the pulmonary artery: intermediate results of coronary elongation. AB - A two coronary system is preferred for correcting anomalous left coronary artery from the pulmonary artery (ALCAPA); however, translocation is not always possible. In countries where neonatal arterial switch operations have not been perfected coronary transfer can be difficult. The purpose of this report is to describe the intermediate results using the coronary elongation and translocation technique in developing countries. Records of patients undergoing operation by the International Children's Heart Foundation team were reviewed (April 1993 October 2008) for those undergoing ALCAPA repair. All patients received a 2-D echocardiographic-color Doppler examination prior to discharge and at follow-up. A total of 13 patients were identified, age ranged from 9 days to 41 years. All but one patient were operated upon at one of our affiliate hospitals in Croatia, Belarus, China and Colombia. All patients presented with moderate to severe mitral regurgitation and cardiac failure. Follow-up ranged from six months to 9.5 years postoperatively. Color Doppler showed a patent left coronary artery; echocardiography estimated a normal left ventricular ejection fraction and improved mitral regurgitation in all patients. The technique provides an alternative approach to translocation for ALCAPA in countries where routine neonatal coronary transfer techniques may not be perfected. Intermediate results are comparable to translocation. PMID- 19679611 TI - Early calcification of the aortic Mitroflow pericardial bioprosthesis in the elderly. AB - BACKGROUND: We report our experience in the elderly with aortic valve replacement using the Mitroflow A12 pericardial bioprosthesis. METHODS: From January 1993 to January 2006, 491 patients over the age of 70 years received an aortic Mitroflow A12 bioprosthesis implantation. Concomitant procedures included coronary artery bypass grafting in 20% of patients. All patients had routine postoperative Echo Doppler studies at discharge, one month and a mean of 11.1 months after surgery and annually thereafter. RESULTS: Twenty (4%) patients underwent a second aortic valve replacement due to bioprosthetic valve dysfunction (Group 2). Calcified stenosis was the most common finding at reoperation (98%). Median time to valve reoperation was 76 months. Of patients requiring reoperation, median age at first and second implantation was 73 (70-78) and 79 (76-83) years, respectively. For all patients, freedom from structural valve dysfunction (SVD) was 95+/-3% at 5 years and 55.8+/-2% at 10 years. Bioprosthetic valve deterioration was identified in 27 patients (Group 1). Median age of these patients at first operation and at diagnosis of deterioration by echo was 75 (70-84) and 77 (70-82) years, respectively. The median interval between operation and detection of bioprosthesis valve deterioration was 46 months. Among the total patient population, freedom from bioprosthetic deterioration was 85.7+/-2% at 5 years and 33.5+/-4% at 10 years. CONCLUSION: The Mitroflow A12 pericardial bioprosthesis provides less than optimal performance in elderly patients. PMID- 19679612 TI - Herpes zoster ophthalmicus. PMID- 19679613 TI - Hormonal contraception and risk of venous thromboembolism: national follow-up study. AB - OBJECTIVE: To assess the risk of venous thrombosis in current users of different types of hormonal contraception, focusing on regimen, oestrogen dose, type of progestogen, and route of administration. DESIGN: National cohort study. SETTING: Denmark, 1995-2005. PARTICIPANTS: Danish women aged 15-49 with no history of cardiovascular or malignant disease. MAIN OUTCOME MEASURES: Adjusted rate ratios for all first time deep venous thrombosis, portal thrombosis, thrombosis of caval vein, thrombosis of renal vein, unspecified deep vein thrombosis, and pulmonary embolism during the study period. RESULTS: 10.4 million woman years were recorded, 3.3 million woman years in receipt of oral contraceptives. In total, 4213 venous thrombotic events were observed, 2045 in current users of oral contraceptives. The overall absolute risk of venous thrombosis per 10 000 woman years in non-users of oral contraceptives was 3.01 and in current users was 6.29. Compared with non-users of combined oral contraceptives the rate ratio of venous thrombembolism in current users decreased with duration of use (<1 year 4.17, 95% confidence interval 3.73 to 4.66, 1-4 years 2.98, 2.73 to 3.26, and >4 years 2.76, 2.53 to 3.02; P<0.001) and with decreasing dose of oestrogen. Compared with oral contraceptives containing levonorgestrel and with the same dose of oestrogen and length of use, the rate ratio for oral contraceptives with norethisterone was 0.98 (0.71 to 1.37), with norgestimate 1.19 (0.96 to 1.47), with desogestrel 1.82 (1.49 to 2.22), with gestodene 1.86 (1.59 to 2.18), with drospirenone 1.64 (1.27 to 2.10), and with cyproterone 1.88 (1.47 to 2.42). Compared with non-users of oral contraceptives, the rate ratio for venous thromboembolism in users of progestogen only oral contraceptives with levonorgestrel or norethisterone was 0.59 (0.33 to 1.03) or with 75 mug desogestrel was 1.12 (0.36 to 3.49), and for hormone releasing intrauterine devices was 0.90 (0.64 to 1.26). CONCLUSION: The risk of venous thrombosis in current users of combined oral contraceptives decreases with duration of use and decreasing oestrogen dose. For the same dose of oestrogen and the same length of use, oral contraceptives with desogestrel, gestodene, or drospirenone were associated with a significantly higher risk of venous thrombosis than oral contraceptives with levonorgestrel. Progestogen only pills and hormone releasing intrauterine devices were not associated with any increased risk of venous thrombosis. PMID- 19679614 TI - The venous thrombotic risk of oral contraceptives, effects of oestrogen dose and progestogen type: results of the MEGA case-control study. AB - OBJECTIVE: To assess the thrombotic risk associated with oral contraceptive use with a focus on dose of oestrogen and type of progestogen of oral contraceptives available in the Netherlands. DESIGN: Population based case-control study. SETTING: Six participating anticoagulation clinics in the Netherlands (Amersfoort, Amsterdam, The Hague, Leiden, Rotterdam, and Utrecht). PARTICIPANTS: Premenopausal women <50 years old who were not pregnant, not within four weeks postpartum, and not using a hormone excreting intrauterine device or depot contraceptive. Analysis included 1524 patients and 1760 controls. MAIN OUTCOME MEASURES: First objectively diagnosed episodes of deep venous thrombosis of the leg or pulmonary embolism. Odds ratios calculated by cross-tabulation with a 95% confidence interval according to Woolf's method; adjusted odds ratios estimated by unconditional logistic regression, standard errors derived from the model. RESULTS: Currently available oral contraceptives increased the risk of venous thrombosis fivefold compared with non-use (odds ratio 5.0, 95% CI 4.2 to 5.8). The risk clearly differed by type of progestogen and dose of oestrogen. The use of oral contraceptives containing levonorgestrel was associated with an almost fourfold increased risk of venous thrombosis (odds ratio 3.6, 2.9 to 4.6) relative to non-users, whereas the risk of venous thrombosis compared with non use was increased 5.6-fold for gestodene (5.6, 3.7 to 8.4), 7.3-fold for desogestrel (7.3, 5.3 to 10.0), 6.8-fold for cyproterone acetate (6.8, 4.7 to 10.0), and 6.3-fold for drospirenone (6.3, 2.9 to 13.7). The risk of venous thrombosis was positively associated with oestrogen dose. We confirmed a high risk of venous thrombosis during the first months of oral contraceptive use irrespective of the type of oral contraceptives. CONCLUSIONS: Currently available oral contraceptives still have a major impact on thrombosis occurrence and many women do not use the safest brands with regard to risk of venous thrombosis. PMID- 19679615 TI - Alarm symptoms and identification of non-cancer diagnoses in primary care: cohort study. AB - OBJECTIVE: To evaluate the predictive value of alarm symptoms for specified non cancer diagnoses and cancer diagnoses in primary care. DESIGN: Cohort study using the general practice research database. SETTING: 128 general practices in the UK contributing data, 1994-2000. PARTICIPANTS: 762 325 patients aged 15 or older. MAIN OUTCOME MEASURES: Up to 15 pre-specified, non-cancer diagnoses associated with four alarm symptoms (haematuria, haemoptysis, dysphagia, rectal bleeding) at 90 days and three years after the first recorded alarm symptom. For each outcome analyses were implemented separately in a time to event framework. Data were censored if patients died, left the practice, or reached the end of the study period. RESULTS: We analysed data on first episodes of haematuria (11 108), haemoptysis (4812), dysphagia (5999), or rectal bleeding (15 289). Non-cancer diagnoses were common in patients who presented with alarm symptoms. The proportion diagnosed with either cancer or non-cancer diagnoses generally increased with age. In patients presenting with haematuria, the proportions diagnosed with either cancer or non-cancer diagnoses within 90 days were 17.5% (95% confidence interval 16.4% to 18.6%) in women and 18.3% (17.4% to 19.3%) in men. For the other symptoms the proportions were 25.7% (23.8% to 27.8%) and 24% (22.5% to 25.6%) for haemoptysis, 17.2% (16% to 18.5%) and 22.6% (21% to 24.3%) for dysphagia, and 14.5% (13.7% to 15.3%) and 16.7% (15.8% to 17.5%) for rectal bleeding. CONCLUSION: Clinically relevant diagnoses are made in a high proportion of patients presenting with alarm symptoms. For every four to seven patients evaluated for haematuria, haemoptysis, dysphagia, or rectal bleeding, relevant diagnoses will be identified in one patient within 90 days. PMID- 19679616 TI - Disagreements in meta-analyses using outcomes measured on continuous or rating scales: observer agreement study. AB - OBJECTIVE: To study the inter-observer variation related to extraction of continuous and numerical rating scale data from trial reports for use in meta analyses. DESIGN: Observer agreement study. DATA SOURCES: A random sample of 10 Cochrane reviews that presented a result as a standardised mean difference (SMD), the protocols for the reviews and the trial reports (n=45) were retrieved. DATA EXTRACTION: Five experienced methodologists and five PhD students independently extracted data from the trial reports for calculation of the first SMD result in each review. The observers did not have access to the reviews but to the protocols, where the relevant outcome was highlighted. The agreement was analysed at both trial and meta-analysis level, pairing the observers in all possible ways (45 pairs, yielding 2025 pairs of trials and 450 pairs of meta-analyses). Agreement was defined as SMDs that differed less than 0.1 in their point estimates or confidence intervals. RESULTS: The agreement was 53% at trial level and 31% at meta-analysis level. Including all pairs, the median disagreement was SMD=0.22 (interquartile range 0.07-0.61). The experts agreed somewhat more than the PhD students at trial level (61% v 46%), but not at meta-analysis level. Important reasons for disagreement were differences in selection of time points, scales, control groups, and type of calculations; whether to include a trial in the meta-analysis; and data extraction errors made by the observers. In 14 out of the 100 SMDs calculated at the meta-analysis level, individual observers reached different conclusions than the originally published review. CONCLUSIONS: Disagreements were common and often larger than the effect of commonly used treatments. Meta-analyses using SMDs are prone to observer variation and should be interpreted with caution. The reliability of meta-analyses might be improved by having more detailed review protocols, more than one observer, and statistical expertise. PMID- 19679617 TI - Oral contraceptives and venous thromboembolism. PMID- 19679618 TI - Prostate specific antigen screening. PMID- 19679619 TI - Aging and cancer: converging routes to disease prevention. AB - An integrative approach to the combined challenges of aging, cancer and stress is a necessary part of a global vision of wellness. Recent research into the mechanisms of aging, cancer and stress has established the biological links between these processes. Understanding these links is an important stepping-stone for developing approaches and therapies that ensure wellness throughout all stages of aging. This paper will consider the most recent developments in research into the molecular mechanisms common to aging and cancer and will discuss the effectiveness of natural approaches for preventing disease. Metabolic regulators as well as nutrient and energy sensors are involved in the processes of aging and cancer, and these are open to external manipulation and control. It is now becoming possible to demonstrate how nutrition, physical activity and stress control can lead to disease-free aging. PMID- 19679621 TI - The effects of an exercise program in leukemia patients. AB - PURPOSE: To examine the feasibility of administering an in-hospital exercise program to acute leukemia patients undergoing chemotherapy. A secondary purpose explored the impact of exercise on selected physiological, psychological, and inflammatory markers. METHODS: Ten patients, aged 18 to 50 years, diagnosed with acute leukemia or newly relapsed were assessed for body weight, height, body composition (skinfolds), cardiorespiratory endurance (total minutes on bicycle ergometer at 60% heart rate reserve), dynamic muscular endurance (Rocky Mountain Cancer Rehabilitation Institute protocol), fatigue (Revised Piper Fatigue Scale), depression (Center for Epidemiologic Studies Depression scale, National Institute of Mental Health questionnaire), and quality of life (Functional Assessment of Cancer Therapy-General) at baseline (within 3 days of diagnosis) and at the end of induction phase of treatment. Blood draws were taken at baseline, midpoint, and at the end of induction for analyses of inflammatory markers (Linco Luminex assay). Combined aerobic and strength training exercises were administered 3 times per week, twice daily, for 30 minutes. Paired-samples t-tests were used for the analyses of physiological and psychological parameters. One-way repeated measures analysis of variance was used for the analyses of inflammatory markers. RESULTS: Significant improvements in cardiorespiratory endurance (P = .009, baseline 8.9 +/- 8.8 minutes, postexercise intervention 17 +/- 14.3 minutes) with significant reductions in total fatigue scores (P = .009, baseline 4.6 +/- 1.7, postexercise intervention 1.8 +/- 1.6) and depression scores (P = .023, baseline 19 +/- 11.5, postexercise intervention 12 +/- 8.2) were observed. Marginally significant decrease in interleukin-6 (IL-6; P = .059) with no significant changes in IL-10 (P = .223) or interferon-gamma (P = .882) were observed. CONCLUSION: Administration of exercise to acute leukemia patients undergoing treatment is feasible. The exercise protocol used increased cardiovascular endurance, reduced fatigue and depression scores, and maintained quality of life. Although no significant change in inflammation was observed, a trend demonstrating a reduction in IL-6 and an increase in IL-10 warrants further investigation. PMID- 19679620 TI - Feasibility trial of electroacupuncture for aromatase inhibitor--related arthralgia in breast cancer survivors. AB - BACKGROUND: Arthralgia affects postmenopausal women receiving aromatase inhibitors (AIs) for breast cancer. Given the existing evidence for electroacupuncture (EA) for treatment of osteoarthritis in the general population, this study aims to establish the feasibility of studying EA for treating AI-related arthralgia. PATIENTS AND METHODS: Postmenopausal women with stage I-III breast cancer who reported AI-related arthralgia were enrolled in a single-arm feasibility trial. EA was provided twice a week for 2 weeks followed by 6 weekly treatments. The protocol was based on Chinese medicine diagnosis of "Bi" syndrome with electrostimulation of needles around the painful joint(s). Pain severity of the modified Brief Pain Inventory was used as the primary outcome. Joint stiffness, joint interference, and Patient Global Impression of Change (PGIC) were secondary outcomes. Paired t tests were used for analysis. RESULTS: Twelve women were enrolled and all provided data for analysis. From baseline to the end of intervention, patients reported reduction in pain severity (from 5.3 to 1.9), stiffness (from 6.9 to 2.4), and joint symptom interference (from 4.7 to 0.8), all P < .001; 11/12 considered joint symptoms "very much better" based on the PGIC. Subjects also reported significant decrease in fatigue (from 4.4 to 1.9, P = .005) and anxiety (from 7.1 to 4.8, P = .01). No infection or development or worsening of lymphedema was observed. CONCLUSION: Preliminary data establish the feasibility of recruitment and acceptance as well as promising preliminary safety and effectiveness. A randomized controlled trial is warranted to establish the efficacy of EA for AI-related arthralgia in breast cancer survivors. PMID- 19679622 TI - Aerobic exercise intensity in breast cancer patients: a preliminary investigation. AB - PURPOSE: This study compared the heart rate (HR), rate of perceived exertion (RPE), and blood lactate (BL) responses to aerobic exercise between posttreated breast cancer patients and apparently healthy, age-matched controls. METHODS: Seven patients and 7 control subjects underwent a submaximal treadmill test for the estimation of Vo(2max). Exercise intensities of 40%, 60%, and 70% of Vo(2max) were calculated from the treadmill test and randomly examined between groups during three 9-minute exercise bouts on 3 different days. Independent samples t tests were used to examine the HR, RPE, and BL responses at each intensity between groups. RESULTS: No significant differences were observed between the control and patient groups for HR, RPE, and BL at 40% (101 +/- 9 vs 101 +/- 11 bpm, P = .979; 8 +/- 1 vs 9 +/- 3, P = .237; and 1.11 +/- 0.73 vs 1.26 +/- 0.64 mmol/L, P = .188, respectively) and 60% (127 +/- 17 vs 117 +/- 13 bpm, P = .523; 12 +/- 2 vs 11 +/- 3, P = .267, and 3.83 +/- 2.48 vs 2.23 +/- 1.65 mmol/L, P = .237, respectively) of Vo(2max) . At 70% of Vo(2max), no significant differences were found for HR (151 +/- 27 vs 135 +/- 13 bpm, P = .704) and RPE (14 +/- 1 vs 13 +/- 3, P = .181), but lower BL responses were observed in the patient group (7.70 +/- 1.62 vs 3.29 +/- 1.08 mmol/L, P < .0005). CONCLUSIONS: The results suggest similar HR, RPE, and BL responses between patients and control subjects at 40%, 60%, and 70% of Vo(2max) , except for BL at 70% of Vo(2max). The lower BL response in the patient group at 70% of Vo(2max) was somewhat unexpected. Further research is needed to confirm or refute the results of this study to allow for a clearer understanding of the physiological responses of breast cancer patients to aerobic exercise at moderate or higher intensities so that safer aerobic exercise prescriptions can be developed for this population. PMID- 19679623 TI - Impact of standardized Rhus verniciflua stokes extract as complementary therapy on metastatic colorectal cancer: a Korean single-center experience. AB - BACKGROUND: To investigate the clinical feasibility of the standardized Rhus verniciflua Stokes (RVS) extract for the metastatic colorectal cancer (mCRC), experimentally proven to have anticancer activities. PATIENTS AND METHODS: From July 2006 to November 2007, patients with conventional chemotherapy refractory mCRC were checked. After fulfilling inclusion/exclusion criteria, 36 patients were eligible for the final analysis. Overall survival and adverse events of patients treated with RVS in the aftercare period were determined. RESULTS: On October 21, 2008, a total of 26 patients died while the remaining 10 patients were alive with evidence of disease. The median RVS administration period was 2.7 months (95% confidence interval, 1.9-3.5). The median overall survival for the entire population was 10.9 months (95% confidence interval, 5.6-16.1) and 1-year survival rate was 44.4%, which is compatible with external controls. By survival analysis using Cox proportional hazards model, the performance status and the prior chemotherapy regimen number significantly affected overall survival. Adverse reactions to the RVS treatment were mostly mild and self-limiting. CONCLUSION: Complementary treatment with the standardized RVS extract might be beneficial for patients with mCRC, since it positively affected overall survival without significant side effects. This study suggests that RVS could be a natural anticancer agent candidate for the treatment of colorectal adenocarcinoma. PMID- 19679624 TI - What is an exceptional cancer trajectory?: Multiple stakeholder perspectives on cancer trajectories in relation to complementary and alternative medicine use. AB - BACKGROUND: Although links between exceptional cancer trajectories (ECTs) and complementary and alternative medicines (CAMs) have been suggested, most research on ECT uses predefined criteria for ECTs without necessarily including CAM use. Little knowledge can be found about subjective perspectives of ECTs in relation to CAM. OBJECTIVES: This Swedish study explores how patients, significant others, and CAM and biomedical health care (BHC) providers reason about ECT, including the role of CAM. METHODS: Using a case study approach, open interviews were conducted with multiple stakeholders about cancer trajectories reported as exceptional. RESULTS: In-depth analysis of 5 cases indicated that although all stakeholders agreed that the reported cases were positive, in no case was the same trajectory considered exceptional by all stakeholders. Different stakeholders shared neither conceptualizations of what constituted an ECT nor attributions for positive trajectories. Patients, significant others, and CAM providers emphasized unexpectedly long survival and well-being in ECTs, whereas BHC providers only referred to survival. CAM and BHC providers normalized reported trajectories, although the forms for this differed between provider groups. CONCLUDING DISCUSSION: Stakeholders' accounts are discussed relative to different health care domains, legitimization processes, and legal constraints, which have led to the lack of a public sphere for CAM use in cancer. PMID- 19679625 TI - An alternative treatment for cervical intraepithelial neoplasia II, III. AB - BACKGROUND: This report describes a case of a woman with progressive and recurrent cervical dysplasia 4 years after cervical conization for severe dysplasia. PATIENT AND METHODS: A 20-year-old female was referred for colposcopy and biopsy following results of moderate to severe atypia of cervical cells on her Papanicolaou (Pap) test. Her colposcopy was satisfactory and her biopsy revealed cervical intraepithelial neoplasia (CIN) II, III. She refused the conventional recommendation of loop electrosurgical excision procedure (LEEP) and, as an alternative, elected to receive escharotic treatment at a frequency of 2 treatments per week for 5 weeks. In addition to the escharotic treatment she followed an oral vitamin and botanical protocol. She was followed for 5 years. RESULTS: The patient's 4-month and 10-month follow-up Pap smears revealed negative cervical cytology for intraepithelial lesion or malignancy. Her 10-month colposcopy was satisfactory and no lesions were noted on the colposcopic exam. Liquid based Pap results continued to remain normal for 5 years after the initiation of treatment. DISCUSSION: Escharotic treatment of high-grade cervical neoplasias with satisfactory colposcopy holds promise as an effective and low risk alternative therapy to LEEP and other excisional procedures. PMID- 19679626 TI - Marked inhibition of growth and invasive parameters of head and neck squamous carcinoma FaDu by a nutrient mixture. AB - Head and neck squamous cell carcinomas (HNSCCs) are known for their aggressive growth and propensity to metastasize. The authors investigated the effect of a novel nutrient mixture (NM) containing ascorbic acid, lysine, proline, and green tea extract on human HNSCC cell line FaDu in vivo and in vitro. Athymic male nude mice (n = 12) were inoculated with 3 x 10(6) FaDu cells subcutaneously and randomly divided into 2 groups: group A was fed a regular diet and group B a regular diet supplemented with 0.5% NM. Four weeks later, the mice were sacrificed and their tumors were excised, weighted, and processed for histology. In vitro, FaDu cells were cultured in Dulbecco's modified Eagle's medium and exposed to NM at 0 to 1000 microg/mL in triplicate. Cell proliferation was assessed by MTT assay, matrix metalloproteinase (MMP) secretion by gelatinase zymography, invasion through Matrigel, apoptosis by live-green caspases, and cell morphology by hematoxylin-eosin staining. NM inhibited the growth of tumors by 55% (P = .0002) and exhibited dose-dependent toxicity on FaDu cells in vitro, with 53% (P = .0003) at 1000 microg/mL NM. Zymography revealed MMP-2 and phorbol 12-myristate 13-acetate-induced MMP-9 secretion. NM inhibited secretion of both MMPs in a dose-dependent manner, with virtual total inhibition at 1000 microg/mL. NM significantly inhibited FaDu invasion through Matrigel with total block at 1000 microg/mL. NM induced dose-dependent apoptosis. In conclusion, NM has therapeutic potential in the treatment of HNSCC by significantly suppressing tumor growth and significantly inhibiting MMP secretion and invasion of HNSCC cells in vitro. PMID- 19679627 TI - Potency of andrographolide as an antitumor compound in BHC-induced liver damage. AB - RELEVANCE: The present investigation relates to the influence of andrographolide, an active compound of Andrographis paniculata Nees. It reverses an experimental liver carcinogenic condition of mice to normal and might be a potential therapeutic/preventive agent for human liver cancer. OBJECTIVE: A. paniculata (Kalmegh) is extensively used in the Indian traditional system of medicine as a hepatoprotective and hepatostimulative agent and has been reported to have protective effect against different hepatotoxins. MATERIALS AND METHODS: Histomorphological, ultrastructural, and biochemical studies were performed for the effect of the andrographolide on control mice, mice treated with hexachlorocyclohexane (BHC) only and BHC + andrographolide. Enzymes for liver function tests were analyzed by spectrophotometric method. RESULTS: The BHC experimental model forms an irreversible liver tumor in male mice. The histological and ultrastructural changes observed in andrographolide supplementation emphasize the recovery of the damaged liver. This recovery was also reflected in the neoplastic nodule formation. The activity of phosphorylase and glucose-6-phosphatase in the liver of the andrographolide-supplemented group suggests improved glycogenolysis in liver. Serum glutamate pyruvate transaminase, serum glutamate oxalate transaminase, alkaline phosphatase, acid phosphatase, and gamma-glutamyl transpeptidase showed a significant decrease in andrographolide supplemented animals as compared with BHC-treated animals, suggesting regenerative effects elicited by andrographolide. CONCLUSION: The study indicates that the regenerative capability elicited by andrographolide is possibly due to its ability to reactivate liver function enzymes that catalyze the reaction of several biochemical and synthetic processes and that it may be useful for severe liver damage conditions. PMID- 19679628 TI - False-negative results from point-of-care qualitative human chorionic gonadotropin (hCG) devices caused by excess hCGbeta core fragment vary with device lot number. PMID- 19679629 TI - Simple and reliable measurement of nitric oxide metabolites in plasma. PMID- 19679630 TI - Short- and long-term risk stratification using a next-generation, high sensitivity research cardiac troponin I (hs-cTnI) assay in an emergency department chest pain population. AB - BACKGROUND: The next-generation, high-sensitivity cardiac troponin assays can measure quantifiable concentrations of cTn in a majority of individuals, but there are few studies assessing these assays for risk stratification. The present study was undertaken to determine if a research hs-cTnI assay can be useful for predicting death/myocardial infarction (MI), both short- and long-term, in an emergency department acute coronary syndrome (ACS) population. METHODS: In a cohort of 383 subjects, originally recruited in 1996, presenting to the emergency department with symptoms suggestive of ACS, the heparin plasma obtained at initial presentation was thawed and measured in 2007 with a research hs-cTnI assay. AccuTnI (Beckman Coulter) measurements were made on these same samples in 2003. The population was divided into 4 groups by hs-cTnI: <5.00, 5.00-9.99, 10.00-40.00, and >40.00 ng/L. Kaplan-Meier, Cox proportional hazards, ROC curves, and logistic regression analyses were used to identify which hs-cTnI concentrations were predictive of death/MI within 10 years after presentation. RESULTS: There were significant differences between the hs-cTnI groups for the probability of death/MI up to 10 years after presentation (P < 0.05). At 6 months, patients with hs-cTnI > or =10.00 ng/L were at higher risk for death/MI (hazard ratio >3.7; P < 0.05) compared with those having hs-cTnI <5.00 ng/L. ROC curve analysis for death/MI at 30 days with the hs-cTnI assay had an area under the curve of 0.74 (95% CI 0.65-0.82), with logistic models yielding an optimal assay threshold of 12.68 ng/L. CONCLUSIONS: This research hs-cTnI assay appears useful for risk stratification for death/MI in an ACS population. PMID- 19679631 TI - Interactive modeling for ongoing utility of pharmacogenetic diagnostic testing: application for warfarin therapy. AB - BACKGROUND: The application of pharmacogenetic results requires demonstrable correlations between a test result and an indicated specific course of action. We developed a computational decision-support tool that combines patient-specific genotype and phenotype information to provide strategic dosage guidance. This tool, through estimating quantitative and temporal parameters associated with the metabolism- and concentration-dependent response to warfarin, provides the necessary patient-specific context for interpreting international normalized ratio (INR) measurements. METHODS: We analyzed clinical information, plasma S warfarin concentration, and CYP2C9 (cytochrome P450, family 2, subfamily C, polypeptide 9) and VKORC1 (vitamin K epoxide reductase complex, subunit 1) genotypes for 137 patients with stable INRs. Plasma S-warfarin concentrations were evaluated by VKORC1 genotype (-1639G>A). The steady-state plasma S-warfarin concentration was calculated with CYP2C9 genotype-based clearance rates and compared with actual measurements. RESULTS: The plasma S-warfarin concentration required to yield the target INR response is significantly (P < 0.05) associated with VKORC1 -1639G>A genotype (GG, 0.68 mg/L; AG, 0.48 mg/L; AA, 0.27 mg/L). Modeling of the plasma S-warfarin concentration according to CYP2C9 genotype predicted 58% of the variation in measured S-warfarin concentration: Measured [S warfarin] = 0.67(Estimated [S-warfarin]) + 0.16 mg/L. CONCLUSIONS: The target interval of plasma S-warfarin concentration required to yield a therapeutic INR can be predicted from the VKORC1 genotype (pharmacodynamics), and the progressive changes in S-warfarin concentration after repeated daily dosing can be predicted from the CYP2C9 genotype (pharmacokinetics). Combining the application of multivariate equations for estimating the maintenance dose with genotype-guided pharmacokinetics/pharmacodynamics modeling provides a powerful tool for maximizing the value of CYP2C9 and VKORC1 test results for ongoing application to patient care. PMID- 19679632 TI - Methylation status and neurodegenerative markers in Parkinson disease. AB - BACKGROUND: Increased concentrations of plasma total homocysteine (tHcy) have been associated with age-related diseases, including dementia, stroke, and Parkinson disease (PD). Methylation status might link Hcy metabolism to neurodegenerative proteins in patients with PD. METHODS: We tested blood samples from 87 patients with PD (median age 68 years; 35 men) for tHcy, methylmalonic acid (MMA), vitamin B(12), vitamin B(6), folate, S-adenosyl methionine (SAM), S adenosyl homocysteine (SAH), and amyloid-beta(1-42). We collected citrate blood from a subset of 45 patients to prepare platelet-rich plasma, and we used washed platelets to prepare cell extracts for amyloid precursor protein (APP) and alpha synuclein assays. We used brain parenchyma sonography to estimate the substantia nigra echogenic area in a subset of 59 patients. RESULTS: Serum concentrations of tHcy were increased in PD patients (median 14.8 micromol/L). tHcy (beta coefficient = -0.276) and serum creatinine (beta = -0.422) were significant predictors of the ratio of SAM/SAH in plasma (P < 0.01). The plasma SAM/SAH ratio was a significant determinant for DemTect scores (beta = 0.612, P = 0.004). Significant negative correlations were found between concentrations of SAH in plasma and platelet APP and between SAM and platelet alpha-synuclein. A larger echogenic area of the substantia nigra was related to higher serum concentrations of MMA (P = 0.016). CONCLUSIONS: Markers of neurodegeneration (APP, alpha synuclein) are related to markers of methylation (SAM, SAH) in patients with PD. Better cognitive function was related to higher methylation potential (SAM/SAH ratio). PMID- 19679634 TI - Analyzing longitudinal data with the linear mixed models procedure in SPSS. AB - Many applied researchers analyzing longitudinal data share a common misconception: that specialized statistical software is necessary to fit hierarchical linear models (also known as linear mixed models [LMMs], or multilevel models) to longitudinal data sets. Although several specialized statistical software programs of high quality are available that allow researchers to fit these models to longitudinal data sets (e.g., HLM), rapid advances in general purpose statistical software packages have recently enabled analysts to fit these same models when using preferred packages that also enable other more common analyses. One of these general purpose statistical packages is SPSS, which includes a very flexible and powerful procedure for fitting LMMs to longitudinal data sets with continuous outcomes. This article aims to present readers with a practical discussion of how to analyze longitudinal data using the LMMs procedure in the SPSS statistical software package. PMID- 19679633 TI - Pediatric brain natriuretic peptide concentrations vary with age and sex and appear to be modulated by testosterone. AB - BACKGROUND: Natriuretic peptide concentrations in adults require age- and sex specific reference intervals for optimal interpretation. Females have higher natriuretic peptide concentrations, and hypotheses suggest that estrogen may be responsible. This study sought to determine the influence of hormone modulation on N-terminal probrain natriuretic peptide (NT-proBNP) by using a pediatric cohort. Children/adolescents typically have rapid hormone changes during puberty, making them an ideal group to study. METHODS: We selected 759 specimens (303 male, 456 female; ages 2 months to 18 years, mean 13 years) obtained from the Mayo Clinic Pediatric Residual Specimen Bank. We measured NT-proBNP, sex hormone binding globulin (SHBG), estradiol, and testosterone by immunoassays or LC-MS/MS and calculated free testosterone. We performed univariate and multivariate analyses to investigate the significance of NT-proBNP with each hormone. RESULTS: Reference values demonstrated a sex difference and sequential age differences in females. Univariate modeling of the hormones with NT-proBNP revealed an independent inverse association of NT-proBNP with testosterone, a direct association with SHBG, and no significant association with estradiol. Multivariate modeling confirmed a strong association of testosterone and SHBG with NT-proBNP. Correlation of hormones with NT-proBNP retained greater significance than either age or sex. CONCLUSIONS: In pediatric patients, NT proBNP is independently associated with both testosterone and SHBG hormone concentrations. Measurements of testosterone are inversely associated with NT proBNP, and estrogens are marginally associated with NT-proBNP in males but not females, suggesting that androgens and not estrogens modulate sex differences notable in natriuretic peptides. Children and adolescents may require an objective assessment of hormones if optimal interpretation of natriuretic peptide concentrations is desired or the concentrations are confounded. . PMID- 19679635 TI - The role of education in improving physicians' professional use of economic evaluations of health interventions: some evidence from a cross-sectional survey in Italy. AB - A cross-sectional survey was carried out on a random sample of Italian physicians through a self-administered questionnaire to describe knowledge, attitudes, and professional behavior toward economic evaluations of health interventions. A response rate of 74.1% was achieved (760 questionnaires). Although many physicians show a positive attitude toward cost-minimization and, to a lesser extent, to cost-effectiveness analysis, they rated their methodological knowledge as unsatisfactory, and the professional use of the economic evaluations of the health interventions in clinical practice is quite low. Multiple logistic regression analysis showed that adequate knowledge and positive attitudes are associated with increased physicians' use of health economic evaluations, as well as time dedicated to continuing medical education and previous training experience about health economics and management. Education and specific training may play an important role in promoting a more cost-conscious behavior of physicians. PMID- 19679636 TI - Medical record review conduction model for improving interrater reliability of abstracting medical-related information. AB - Medical record review (MRR) is often used in clinical research and evaluation, yet there is limited literature regarding best practices in conducting a MRR, and there are few studies reporting interrater reliability (IRR) from MRR data. The aim of this research was twofold: (a) to develop a MRR abstraction tool and standardize the MRR process and (b) to examine the IRR from MRR data. This study introduces the MRR-Conduction Model, which was used to implement a MRR, and examines the IRR between two abstractors who collected preinjury medical and psychiatric, incident-related medical and postinjury head symptom information from the medical records of 47 neurologically injured workers. Results showed that the percentage agreement was > or =85% and the unweighted kappa statistic was > or =.60 for most variables, indicating substantial IRR. An effective and reliable MRR to abstract medical-related information requires planning and time. The MRR-Conduction Model is proposed to guide the process of creating a MRR. PMID- 19679637 TI - Nebulin plays a direct role in promoting strong actin-myosin interactions. AB - The role of the actin filament-associated protein nebulin on mechanical and kinetic properties of the actomyosin motor was investigated in skeletal muscle of wild-type (wt) and nebulin-deficient (nebulin(-)(/)(-)) mice that were 1 d old, an age at which sarcomeric structure is still well preserved. In Ca2+-activated skinned fibers from psoas muscle, we determined the Ca2+ dependence of isometric force and stiffness, the rate of force redevelopment after unloaded shortening (k(TR)), the power during isotonic shortening, and the unloaded shortening velocity (V(0)). Our results show a 65% reduction in isometric force in nebulin( )(/)(-) fibers at saturating [Ca2+], whereas neither thin-filament length nor the Ca2+ sensitivity of the contractile system is affected. Stiffness measurements indicate that the reduction in isometric force is due to a reduction in the number of actin-attached myosin motors, whereas the force of the motor is unchanged. Furthermore, in nebulin(-)(/)(-) fibers, k(TR) is decreased by 57%, V(0) is increased by 63%, and the maximum power is decreased by 80%. These results indicate that, in the absence of nebulin, the attachment probability of the myosin motors to actin is decreased, revealing a direct role for nebulin in promoting strong actomyosin interactions responsible for force and power production. PMID- 19679638 TI - Divalent metal transporter 1 is involved in amyloid precursor protein processing and Abeta generation. AB - The amyloid-beta precursor protein (APP) and its pathogenic byproduct beta amyloid peptide (Abeta) play central roles in the pathogenesis of Alzheimer's disease (AD). Reduction in levels of the potentially toxic Abeta is one of the most important therapeutic goals in AD. Recent studies have shown that bivalent metals such as iron, copper, and zinc are involved in APP expression, Abeta deposition, and senile plaque formation in the AD brain. However, the underlying mechanisms involved in abnormal homeostasis of bivalent metals in AD brain remain unclear. In the present study, we found that two isoforms of the divalent metal transporter 1 (DMT1), DMT1-IRE, and DMT1-nonIRE, were colocalized with Abeta in the plaques of postmortem AD brain. Using the APP/PS1 transgenic mouse model, we found that the levels of both DMT1-IRE and DMT1-nonIRE were significantly increased in the cortex and hippocampus compared with wild type-control. We further verified the proposed mechanisms by which DMT1 might be involved in APP processing and Abeta secretion by using the SH-SY5Y cell line stably overexpressing human APP Swedish mutation (APPsw) as a cell model. We found that overexpression of APPsw resulted in increased expression levels of both DMT1-IRE and DMT1-nonIRE in SH-SY5Y cells. Interestingly, silencing of endogenous DMT1 by RNA interference, which reduced bivalent ion influx, led to reductions of APP expression and Abeta secretion. These findings suggest both that DMT1 plays a critical role in ion-mediated neuropathogenesis in AD and that pharmacological blockage of DMT1 may provide novel therapeutic strategies against AD. PMID- 19679640 TI - Dichloroacetate treatment partially regresses established pulmonary hypertension in mice with SM22alpha-targeted overexpression of the serotonin transporter. AB - Voltage-gated potassium (Kv)1.5 is decreased in pulmonary arteries (PAs) of patients with idiopathic pulmonary arterial hypertension (IPAH) and in experimental models including mice with SM22alpha-targeted overexpression of the serotonin transporter (5-HTT). The mechanisms underlying these abnormalities, however, remain unknown. Dichloroacetate (DCA) inhibits chronic hypoxia- or monocrotaline-induced PAH by inhibiting nuclear factor of activated T-cells (NFAT)c2 and increasing Kv1.5. Therefore, we hypothesized that DCA could regress established PAH in SM22-5-HTT+ mice. We evaluated pulmonary hemodynamics, vascular remodeling, NFATc2, and Kv1.5 protein in 20-wk-old SM22-5-HTT+ or wild type mice treated for 1, 7, and 21 d with DCA, cyclosporine-A (NFAT inhibitor), or vehicle. DCA partially reversed PAH in SM22-5-HTT+ mice by decreasing proliferation and increasing apoptosis in muscularized PAs. Furthermore, serotonin (10(-8)-10(-6) M) dose-dependently increased PA-smooth muscle cell (PA SMC) proliferation in culture (EC(50)=0.97 x 10(-7) M) and DCA (5 x 10(-4) M) vs. PBS markedly reduced the growth of PA-SMC from IPAH and control patients treated with the highest dose of serotonin by 50 and 30%, respectively. Finally, although serotonin induces NFATc2 activation in PA-SMCs, inhibition of NFATc2 alone with cyclosporine-A was not sufficient for reversing PAH in this model. Our results support the possibility that DCA may be an interesting agent for investigation in patients with PAH. PMID- 19679639 TI - Biomarkers of morphine tolerance and dependence are prevented by morphine-induced endocytosis of a mutant mu-opioid receptor. AB - Growing evidence shows that trafficking of the mu-opioid receptor (MOR) is a critical process in functional recovery from desensitization following activation and plays important roles in morphine tolerance and dependence largely because of the failure of morphine to promote such trafficking. However, morphine tolerance and dependence are believed to be mediated by multiple mechanisms, including well documented biochemical changes in cAMP activity, N-methyl-D-aspartate receptors (NMDARs), glucocorticoid receptors (GRs), and c-fos. Here, we assess the consequences of promoting morphine-induced endocytosis on these biochemical changes utilizing a knock-in mouse model, RMOR, in which MORs undergo morphine induced endocytosis. Chronic morphine treatment of wild-type (WT) mice promoted superactivation of adenylyl cyclase, alterations in NMDARs, and up-regulation of GR and c-fos in distinct brain regions. Notably, none of these biochemical changes occurred in the RMOR-knock-in mice. Together, these data demonstrate that morphine tolerance and dependence are mediated by multiple biochemical mechanisms and that MOR endocytosis plays a critical role in each of these mechanisms. PMID- 19679641 TI - Long-term follow-up of 1036 cerebral aneurysms treated by bare coils: a multicentric cohort treated between 1998 and 2003. AB - BACKGROUND AND PURPOSE: The endovascular treatment (EVT) of cerebral aneurysms has experienced a revolution since 1991 with the introduction of platinum coil technology. During the past 10 years, there has been significant study of the feasibility of this technique, and clinical results of EVT have been published. The long-term durability of Guglielmi detachable coil (GDC) embolization of cerebral aneurysms still remains unknown. The purpose of this study was to evaluate the stability of anatomic occlusion of aneurysms and to assess the rate of recanalization and retreatment of these aneurysms. MATERIALS AND METHODS: Between January 1998 and December 2003, 1036 aneurysms (804 ruptured and 232 nonruptured) were treated consecutively with GDC coils in 5 neuroradiology centers. Procedural feasibility, acute angiographic occlusion results, morbidity, and mortality associated with this technique were assessed. All patients were regularly followed by digital subtraction angiography and MR imaging each year after treatment. RESULTS: Initial acute angiographic results in 1036 aneurysms demonstrated total occlusion in 731 patients (70.5%), subtotal occlusion in 252 (24.3%), incomplete occlusion in 20 (1.9%), and failures in 33% (3.3%) aneurysms. A remodeling technique was used in 10%. A second procedure was performed for 72 aneurysms (7%). The total aneurysm follow-up time was 49,923 months (4160.25 aneurysm-years). The retreatment period was either in the months following initial treatment in aneurysms incompletely occluded or in years due to recanalization or de novo aneurysms. Fewer than 5 patients rebled during 10 years of follow-up. Long-term follow-up angiograms were obtained in 899 aneurysms, with 646 total, 230 subtotal, and 23 incomplete results. CONCLUSIONS: Long-term follow up of cerebral aneurysms is necessary to depict recanalization. Only 7% of the aneurysms were retreated. Use of bare coils gives a good long-term level of occlusion. PMID- 19679643 TI - RIP kinases initiate programmed necrosis. AB - Some lethal stimuli can induce either apoptosis or necrosis, depending on the cell type and/or experimental setting. Until recently, the molecular bases of this phenomenon were largely unknown. Now, two members of the receptor interacting serine-threonine kinase (RIP) family, RIP1 and RIP3, have been demonstrated to control the switch between apoptotic and necrotic cell death. Some mechanistic details, however, remain controversial. PMID- 19679644 TI - Macrophage metalloelastase: stretching therapeutic opportunities. AB - While tissue macrophages are at the first line of microbial host defense, they are also convenient hideouts for pathogens escaping immune attack. Houghton et al. discovered that alveolar macrophage mobilizes macrophage metalloelastase to destroy bacteria present inside the cell. PMID- 19679642 TI - RNA polymerase III can drive polycistronic expression of functional interfering RNAs designed to resemble microRNAs. AB - In both research and therapeutic applications of RNA interference, it is often advantageous to silence several targets simultaneously. Toward this end, several groups have developed vectors that utilize the model of endogenously encoded micro (mi) RNAs, where a single RNA polymerase II promoter can drive the expression of multiple interfering RNAs. Stronger pol III promoters have been used to drive individual short hairpin (sh) RNAs, but to date, it has been necessary to repeat the promoter in each silencing cassette to achieve multiplexed expression from a single vector. Here, we show that it is possible to drive polycistronic expression from a single pol III promoter when the interfering RNAs are formatted to resemble miRNAs rather than shRNAs. As many as four miRNAs designed to target hepatitis B virus (HBV) transcripts are shown to be processed and functional in reporter assays as well as in the context of replicating virus in cell culture systems. Although it has been observed that high levels of expression of shRNAs can lead to cytotoxicity, we find no significant evidence in transient transfection assays that the HBV-miRNAs produced by our vectors compete for the activity of endogenously produced miR-122 or for processing of an exogenously expressed miR-EGFP. PMID- 19679645 TI - Electroacupuncture Zusanli (ST36) on Release of Nitric Oxide in the Gracile Nucleus and Improvement of Sensory Neuropathies in Zucker Diabetic Fatty Rats. AB - The purpose of these studies was to examine the effects of electroacupuncture (EA) Zusanli (ST36) on release of nitric oxide (NO) in the gracile nucleus (GN) and determine if functional neuropathic changes were modified by EA ST36-induced NO in the nucleus in Zucker diabetic fatty (ZDF) rats. The foot withdrawal responses to mechanical, thermal and cold stimuli were measured before and after EA stimulation. A microdialysis probe was implanted in the GN and dialysate samples were collected 20 min before, during and after EA ST36. Total nitrate and nitrite (NO(x) (-)) concentrations in the samples were quantified by using chemiluminescence. The baseline dialysate NO(x) (-) concentrations in the GN were decreased in ZDF rats compared to lean control (LC) rats (P < .05). In ZDF rats, dialysate NO(x) (-) releases in the GN were markedly increased during EA ST36, whereas in LC rats, the releases were moderately enhanced at 20-40 min after EA ST36. The withdrawal latencies to mechanical, cold and thermal stimuli were significantly improved 20 min after EA ST36 both in LC and ZDF rats, but not altered by non-acupoint stimulation. The withdrawal latencies to EA ST36 were further potentiated by 3-morpholinyl-sydnoneimine and inhibited by N(G)-Propyl-l arginine infused into the GN in ZDF rats (P < .05). These results show that EA ST36 increases NO release in the GN, and NO in the nucleus modifies withdrawal latencies to mechanical, cold, and thermal nociception stimuli. Data suggest that EA ST36 induces NO release in the GN, which contributes to improvement of sensory neuropathies in rats. PMID- 19679646 TI - Compensation for vertical dysplasia and its clinical application. AB - The purpose of this study was to quantitatively evaluate skeletal and dental compensation in patients with vertical skeletal dysplasias and to determine which dentoalveolar parameters compensate for vertical jaw discrepancies. Cephalometric analyses were performed on pre-treatment lateral cephalographs of 186 orthodontic patients (120 females and 66 males; mean age 15 years 11 months) who met the selection criteria. SN-MP angle was used to classify the facial patterns as: hyperdivergent > 36 degrees, normo = 28-36 degrees, and hypo < 28 degrees. Analysis of variance (ANOVA) was used to determine statistical differences between the means in the three vertical facial types. To evaluate dental compensation quantitatively, correlation analyses were performed to find associations between skeletal and dental parameters. To further elucidate the compensatory nature of the lower incisors, regression analyses and scattergrams were obtained, with SN-MP as a measure of the vertical skeletal discrepancy. ANOVA showed statistically significant differences for most of the skeletal variables, but only for lower incisor height and inclination among the dentoalveolar parameters. Correlation analyses demonstrated significant relationships between various skeletal variables. LI-MP showed a negative relationship with SN-MP, whereas LAMdH demonstrated a positive linear relationship with SN-MP. Among all dentoalveolar heights, UAMxH showed the weakest, and LAMdH the strongest, associations with skeletal parameters. The variability in dentoalveolar compensation therefore demands individualized diagnosis and treatment planning. LAMdH and LI-MP parameters were the most likely, whereas UAMxH was the least likely parameter to compensate for vertical dysplasia. PMID- 19679648 TI - Physical fitness in children with high motor competence is different from that in children with low motor competence. AB - BACKGROUND: Physical therapists often treat children with low motor competence. Earlier studies have demonstrated poor physical fitness outcomes and a reduced level of physical activity for these children compared with their peers with normal motor skills. OBJECTIVE: The aim of this study was to examine how physical fitness developed over time in 2 groups of children: those with a low level of competence in motor skills (low motor competence [LMC]), and those with a high level of competence in motor skills (high motor competence [HMC]). DESIGN AND METHODS: From an initial sample of 67 children, a group of 18 was identified as having HMC or LMC on the Movement Assessment Battery for Children and was selected for the present study. Eight children (3 girls and 5 boys) comprised the LMC group, and 10 children (4 girls and 6 boys) made up the HMC group. A longitudinal design was implemented, and physical fitness in the 2 groups was evaluated by measuring different fitness components over a period of 32 months. RESULTS: A mixed-effects analysis of variance revealed significant main effects for group and for time but no group x time interaction effect. The LMC group performed less well on all physical fitness measures than the HMC group, and both groups scored significantly higher on the physical fitness test after a period of 32 months. The lack of a significant interaction effect indicated that the relative differences in physical fitness outcomes between the groups were relatively constant over time. LIMITATIONS: This study was limited by the small sample size and lack of assessment of anthropometric variables and children's perceived self-efficacy. CONCLUSIONS: Children with LMC are likely to have poor physical fitness compared with children with HMC. The differences in physical fitness outcomes between the groups were relatively constant over time. Given that various physical fitness components are linked to different health outcomes, these consequences are matters of concern for both current health status and later health status in children with LMC. PMID- 19679647 TI - Increased telomere fragility and fusions resulting from TRF1 deficiency lead to degenerative pathologies and increased cancer in mice. AB - The telomere repeat-binding factor 1 (TERF1, referred to hereafter as TRF1) is a component of mammalian telomeres whose role in telomere biology and disease has remained elusive. Here, we report on cells and mice conditionally deleted for TRF1. TRF1-deleted mouse embryonic fibroblasts (MEFs) show rapid induction of senescence, which is concomitant with abundant telomeric gamma-H2AX foci and activation of the ATM/ATR downstream checkpoint kinases CHK1 and CHK2. DNA damage foci are rescued by both ATM and ATM/ATR inhibitors, further indicating that both signaling pathways are activated upon TRF1 deletion. Abrogation of the p53 and RB pathways bypasses senescence but leads to chromosomal instability including sister chromatid fusions, chromosome concatenation, and occurrence of multitelomeric signals (MTS). MTS are also elevated in ATR-deficient MEFs or upon treatment with aphidicolin, two conditions known to induce breakage at fragile sites, suggesting that TRF1-depleted telomeres are prone to breakage. To address the impact of these molecular defects in the organism, we deleted TRF1 in stratified epithelia of TRF1(Delta/Delta)K5-Cre mice. These mice die perinatally and show skin hyperpigmentation and epithelial dysplasia, which are associated with induction of telomere-instigated DNA damage, activation of the p53/p21 and p16 pathways, and cell cycle arrest in vivo. p53 deficiency rescues mouse survival but leads to development of squamous cell carcinomas, demonstrating that TRF1 suppresses tumorigenesis. Together, these results demonstrate that dysfunction of a telomere-binding protein is sufficient to produce severe telomeric damage in the absence of telomere shortening, resulting in premature tissue degeneration and development of neoplastic lesions. PMID- 19679650 TI - An appraisal of the performance of the economic and financial crimes commission in Nigeria. AB - This article examines how an anti-graft body, the Economic and Financial Crime Commission (EFCC), has fared in reducing the incidence of corruption in Nigeria, in particular, bank fraud, Internet scam, and bad governance. It first discusses the corruption situation in Nigeria by highlighting public office holders who have been associated with corruption charges. A Likert-type scale is used in designing the questionnaire for data collection. Descriptive and chi-square analyses are used, and results reveal that the performance of the EFCC has been affected by government interference (p < .05). However, although the anti-graft body has not been able to reduce the incidence of bank fraud (p > .05), bad governance and advance fee fraud have recorded appreciable reduction (p < .05). Areas of success as well as challenges that need to be addressed are identified. Specifically, it is recommended that the bill that established EFCC should be amended to reduce government interference and improve its manpower development, especially in the areas of fraud and Internet scam detection. PMID- 19679649 TI - Responses to the cannabinoid receptor-1 antagonist, AM251, are more robust with age and with high-fat feeding. AB - Endocannabinoids (EC) are involved in regulating energy homeostasis, particularly in promoting hyperphagia and the consumption of a palatable diet. We have previously shown that rats given a high-fat (HF) diet display a transient hyperphagia that is normalized by a process partially dependent on leptin. We now propose that the induction of this hyperphagia is mediated, at least partially, by the EC signaling system. Obesity, including diet-induced and age-related, is associated with dysregulation of the EC system, and obese rodent models are hypersensitive to a cannabinoid-1 (CB1) receptor antagonist. This suggests that aged rats will be more responsive to the anorectic effects of a CB1 receptor antagonist. To test this, we examined the responsiveness to CB1 receptor antagonist, AM251, in young and aged rats during two experimental paradigms. First, we administered AM251 simultaneously with the introduction of an HF diet. Second, AM251 treatment began after the establishment of diet-induced obesity. Responses were measured by changes in body weight and composition, calorie intake, serum leptin, and biochemical indicators. The results demonstrated three key findings. 1) CB1 receptor activity contributes to the hyperphagia seen with the introduction of an HF diet. 2) Increased AM251 sensitivity and efficacy is increased with age and HF feeding, with the greatest responsiveness observed in HF-fed, aged rats. 3) AM251 sensitivity is elevated to a greater extent with HF diet than with established obesity. Thus, both age and an HF diet are associated with enhanced anorectic responses to AM251, but the underlying mechanism of these responses remains speculative. PMID- 19679651 TI - Evaluation of tooth-click triggering and speech recognition in assistive technology for computer access. AB - BACKGROUND: Computer access can play an important role in employment and leisure activities following spinal cord injury. The authors' prior work has shown that a tooth-click detecting device, when paired with an optical head mouse, may be used by people with tetraplegia for controlling cursor movement and mouse button clicks. OBJECTIVE: To compare the efficacy of tooth clicks to speech recognition and that of an optical head mouse to a gyrometer head mouse for cursor and mouse button control of a computer. METHODS: Six able-bodied and 3 tetraplegic subjects used the devices listed above to produce cursor movements and mouse clicks in response to a series of prompts displayed on a computer. The time taken to move to and click on each target was recorded. RESULTS: The use of tooth clicks in combination with either an optical head mouse or a gyrometer head mouse can provide hands-free cursor movement and mouse button control at a speed of up to 22% of that of a standard mouse. Tooth clicks were significantly faster at generating mouse button clicks than speech recognition when paired with either type of head mouse device. CONCLUSIONS: Tooth-click detection performed better than speech recognition when paired with both the optical head mouse and the gyrometer head mouse. Such a system may improve computer access for people with tetraplegia. PMID- 19679652 TI - N-myristoylated c-Abl tyrosine kinase localizes to the endoplasmic reticulum upon binding to an allosteric inhibitor. AB - Allosteric kinase inhibitors hold promise for revealing unique features of kinases that may not be apparent using conventional ATP-competitive inhibitors. Here we explore the activity of a previously reported allosteric inhibitor of BCR Abl kinase, GNF-2, against two cellular isoforms of Abl tyrosine kinase: one that carries a myristate in the N terminus and the other that is deficient in N myristoylation. Our results show that GNF-2 inhibits the kinase activity of non myristoylated c-Abl more potently than that of myristoylated c-Abl by binding to the myristate-binding pocket in the C-lobe of the kinase domain. Unexpectedly, indirect immunofluorescence reveals a translocation of myristoylated c-Abl to the endoplasmic reticulum in GNF-2-treated cells, whereas GNF-2 has no detectable effect on the localization of non-myristoylated c-Abl. These results indicate that GNF-2 competes with the NH(2)-terminal myristate for binding to the c-Abl kinase myristate-binding pocket and that the exposed myristoyl group accounts for the localization to the endoplasmic reticulum. We also demonstrate that GNF-2 can inhibit enzymatic and cellular kinase activity of Arg, a kinase highly homologous to c-Abl, which is also likely to be regulated through intramolecular binding of an NH(2)-terminal myristate lipid. These results suggest that non-ATP-competitive inhibitors, such as GNF-2, can serve as chemical tools that can discriminate between c-Abl isoform-specific behaviors. PMID- 19679653 TI - Inhibition of cyclooxygenase-2 down-regulates aromatase activity and decreases proliferation of Leydig tumor cells. AB - Our recent studies have revealed that estrogens stimulate an autocrine mechanism determining Leydig tumor cell proliferation. Estrogen overproduction is due to an elevated steroidogenic factor-1 (SF-1) expression and cAMP-response element binding protein (CREB) phosphorylation, both inducing aromatase overexpression. Although we have shown that increased SF-1 expression depends mainly on higher local insulin-like growth factor I production, the mechanisms and factors determining increased CREB activation in Leydig tumor cells are not completely understood. In this study, we investigated the role of cyclooxygenase-2 (COX-2) in CREB dependent-aromatase expression in Leydig tumor cells. We found that COX-2 is expressed in rat and human Leydigiomas as well as in the rat Leydig tumor cell line R2C, but not in normal testis. Our data indicate that in R2C cells the COX-2 derived prostaglandin E2 (PGE2) binds the PGE2 receptor EP4 and activates protein kinase A (PKA) and ultimately CREB. Inhibitors for COX-2 (NS398), EP4 (AH23848), and PKA (H89) decreased aromatase expression and activity as a consequence of a decreased phosphorylated CREB recruitment to the PII promoter of the aromatase gene. The COX-2/PGE2/PKA pathway also seems to be involved in aromatase post translational activation, an observation that requires further studies. The reduction in aromatase activity was responsible for a drop in estrogen production and subsequent reduction in cyclin E expression resulting in a decrease in tumor Leydig cell proliferation. Furthermore, COX-2 silencing caused a significant decrease in CREB phosphorylation, aromatase expression, and R2C cell proliferation. These novel findings clarify the mechanisms involved in the growth of Leydig cell tumors and should be taken into account in determining new therapeutic approaches. PMID- 19679654 TI - Escherichia coli unsaturated fatty acid synthesis: complex transcription of the fabA gene and in vivo identification of the essential reaction catalyzed by FabB. AB - Although the unsaturated fatty acid (UFA) synthetic pathway of Escherichia coli is the prototype of such pathways, several unresolved issues have accumulated over the years. The key players are the fabA and fabB genes. Earlier studies of fabA transcription showed that the gene was transcribed from two promoters, with one being positively regulated by the FadR protein. The other weaker promoter (which could not be mapped with the technology then available) was considered constitutive because its function was independent of FadR. However, the FabR negative regulator was recently shown to represses fabA transcription. We report that the weak promoter overlaps the FadR-dependent promoter and is regulated by FabR. This promoter is strictly conserved in all E. coli and Salmonella enterica genomes sequenced to date and is thought to provide insurance against inappropriate regulation of fabA transcription by exogenous saturated fatty acids. Also, the fabAup promoter, a mutant promoter previously isolated by selection for increased FabA activity, was shown to be a promoter created de novo by a four-base deletion within the gene located immediately upstream of fabA. Demonstration of the key UFA synthetic reaction catalyzed by FabB has been elusive, although it was known to catalyze an elongation reaction. Strains lacking FabB are UFA auxotrophs indicating that the enzyme catalyzes an essential step in UFA synthesis. Using thioesterases specific for hydrolysis of short chain acyl-ACPs, the intermediates of the UFA synthetic pathway have been followed in vivo for the first time. These experiments showed that a fabB mutant strain accumulated less cis-5-dodecenoic acid than the parental wild-type strain. These data indicate that the key reaction in UFA synthesis catalyzed by FabB is elongation of the cis-3-decenoyl-ACP produced by FabA. PMID- 19679655 TI - Deciphering structural and functional roles of individual disulfide bonds of the mitochondrial sulfhydryl oxidase Erv1p. AB - Erv1p is a FAD-dependent sulfhydryl oxidase of the mitochondrial intermembrane space. It contains three conserved disulfide bonds arranged in two CXXC motifs and one CX(16)C motif. Experimental evidence for the specific roles of the individual disulfide bonds is lacking. In this study, structural and functional roles of the disulfides were dissected systematically using a wide range of biochemical and biophysical methods. Three double cysteine mutants with each pair of cysteines mutated to serines were generated. All of the mutants were purified with the normal FAD binding properties as the wild type Erv1p, showing that none of the three disulfides are essential for FAD binding. Thermal denaturation and trypsin digestion studies showed that the CX(16)C disulfide plays an important role in stabilizing the folding of Erv1p. To understand the functional role of each disulfide, small molecules and the physiological substrate protein Mia40 were used as electron donors in oxygen consumption assays. We show that both CXXC disulfides are required for Erv1 oxidase activity. The active site disulfide is well protected thus requires the shuttle disulfide for its function. Although both mutants of the CXXC motifs were individually inactive, Erv1p activity was partially recovered by mixing these two mutants together, and the recovery was rapid. Thus, we provided the first experimental evidence of electron transfer between the shuttle and active site disulfides of Erv1p, and we propose that both intersubunit and intermolecular electron transfer can occur. PMID- 19679657 TI - The yeast RNA polymerase II-associated factor Iwr1p is involved in the basal and regulated transcription of specific genes. AB - RNA polymerase II (RNA pol II) is a multisubunit enzyme that requires many auxiliary factors for its activity. Over the years, these factors have been identified using both biochemical and genetic approaches. Recently, the systematic characterization of protein complexes by tandem affinity purification and mass spectroscopy has allowed the identification of new components of well established complexes, including the RNA pol II holoenzyme. Using this approach, a novel and highly conserved factor, Iwr1p, that physically interacts with most of the RNA pol II subunits has been described in yeast. Here we show that Iwr1p genetically interacts with components of the basal transcription machinery and plays a role in both basal and regulated transcription. We report that mutation of the IWR1 gene is able to bypass the otherwise essential requirement for the transcriptional regulator negative cofactor 2, which occurs with different components of the basal transcription machinery, including TFIIA and subunits of the mediator complex. Deletion of the IWR1 gene leads to an altered expression of specific genes, including phosphate-responsive genes and SUC2. Our results show that Iwr1p is a nucleocytoplasmic shuttling protein and suggest that Iwr1p acts early in the formation of the pre-initiation complex by mediating the interaction of certain activators with the basal transcription apparatus. PMID- 19679656 TI - Inhibition of prolyl hydroxylase protects against 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine-induced neurotoxicity: model for the potential involvement of the hypoxia-inducible factor pathway in Parkinson disease. AB - Hypoxia-inducible factor (HIF) plays an important role in cell survival by regulating iron, antioxidant defense, and mitochondrial function. Pharmacological inhibitors of the iron-dependent enzyme class prolyl hydroxylases (PHD), which target alpha subunits of HIF proteins for degradation, have recently been demonstrated to alleviate neurodegeneration associated with stroke and hypoxic ischemic injuries. Here we report that inhibition of PHD by 3,4-dihydroxybenzoate (DHB) protects against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) induced nigral dopaminergic cell loss and up-regulates HIF-1alpha within these neurons. Elevations in mRNA and protein levels of HIF-dependent genes heme oxygenase-1 (Ho-1) and manganese superoxide dismutase (Mnsod) following DHB pretreatment alone are also maintained in the presence of MPTP. MPTP-induced reductions in ferroportin and elevations in nigral and striatal iron levels were reverted to levels comparable with that of untreated controls with DHB pretreatment. Reductions in pyruvate dehydrogenase mRNA and activity resulting from MPTP were also found to be attenuated by DHB. In vitro, the HIF pathway was activated in N27 cells grown at 3% oxygen treated with either PHD inhibitors or an iron chelator. Concordant with our in vivo data, the MPP(+)-elicited increase in total iron as well as decreases in cell viability were attenuated in the presence of DHB. Taken together, these data suggest that protection against MPTP neurotoxicity may be mediated by alterations in iron homeostasis and defense against oxidative stress and mitochondrial dysfunction brought about by cellular HIF-1alpha induction. This study provides novel data extending the possible therapeutic utility of HIF induction to a Parkinson disease model of neurodegeneration, which may prove beneficial not only in this disorder itself but also in other diseases associated with metal-induced oxidative stress. PMID- 19679658 TI - Nucleophosmin/B23 regulates ubiquitin dynamics in nucleoli by recruiting deubiquitylating enzyme USP36. AB - The nucleolus is a subnuclear compartment with multiple cellular functions, including ribosome biogenesis. USP36 is a deubiquitylating enzyme that localizes to nucleoli and plays an essential role in regulating the structure and function of the organelle. However, how the localization of USP36 is regulated remains unknown. Here, we identified a short stretch of basic amino acids (RGKEKKIKKFKREKRR) that resides in the C-terminal region of USP36 and serves as a nucleolar localization signal for the protein. We found that this motif interacts with a central acidic region of nucleophosmin/B23, a major nucleolar protein involved in various nucleolar functions. Knockdown of nucleophosmin/B23 resulted in a significant reduction in the amount of USP36 in nucleoli, without affecting the cellular USP36 level. This was associated with elevated ubiquitylation levels of fibrillarin, a USP36 substrate protein in nucleoli. We conclude that nucleophosmin/B23 recruits USP36 to nucleoli, thereby serving as a platform for the regulation of nucleolar protein functions through ubiquitylation/deubiquitylation. PMID- 19679659 TI - Depletion of vitamin E increases amyloid beta accumulation by decreasing its clearances from brain and blood in a mouse model of Alzheimer disease. AB - Increased oxidative damage is a prominent and early feature in Alzheimer disease. We previously crossed Alzheimer disease transgenic (APPsw) model mice with alpha tocopherol transfer protein knock-out (Ttpa(-/-)) mice in which lipid peroxidation in the brain was significantly increased. The resulting double mutant (Ttpa(-/-)APPsw) mice showed increased amyloid beta (Abeta) deposits in the brain, which was ameliorated with alpha-tocopherol supplementation. To investigate the mechanism of the increased Abeta accumulation, we here studied generation, degradation, aggregation, and efflux of Abeta in the mice. The clearance of intracerebral-microinjected (125)I-Abeta(1-40) from brain was decreased in Ttpa(-/-) mice to be compared with wild-type mice, whereas the generation of Abeta was not increased in Ttpa(-/-)APPsw mice. The activity of an Abeta-degrading enzyme, neprilysin, did not decrease, but the expression level of insulin-degrading enzyme was markedly decreased in Ttpa(-/-) mouse brain. In contrast, Abeta aggregation was accelerated in Ttpa(-/-) mouse brains compared with wild-type brains, and well known molecules involved in Abeta transport from brain to blood, low density lipoprotein receptor-related protein-1 (LRP-1) and p glycoprotein, were up-regulated in the small vascular fraction of Ttpa(-/-) mouse brains. Moreover, the disappearance of intravenously administered (125)I-Abeta(1 40) was decreased in Ttpa(-/-) mice with reduced translocation of LRP-1 in the hepatocytes. These results suggest that lipid peroxidation due to depletion of alpha-tocopherol impairs Abeta clearances from the brain and from the blood, possibly causing increased Abeta accumulation in Ttpa(-/-)APPsw mouse brain and plasma. PMID- 19679661 TI - Loss of TMEM16A causes a defect in epithelial Ca2+-dependent chloride transport. AB - Molecular identification of the Ca(2+)-dependent chloride channel TMEM16A (ANO1) provided a fundamental step in understanding Ca(2+)-dependent Cl(-) secretion in epithelia. TMEM16A is an intrinsic constituent of Ca(2+)-dependent Cl(-) channels in cultured epithelia and may control salivary output, but its physiological role in native epithelial tissues remains largely obscure. Here, we demonstrate that Cl(-) secretion in native epithelia activated by Ca(2+)-dependent agonists is missing in mice lacking expression of TMEM16A. Ca(2+)-dependent Cl(-) transport was missing or largely reduced in isolated tracheal and colonic epithelia, as well as hepatocytes and acinar cells from pancreatic and submandibular glands of TMEM16A(-/-) animals. Measurement of particle transport on the surface of tracheas ex vivo indicated largely reduced mucociliary clearance in TMEM16A(-/-) mice. These results clearly demonstrate the broad physiological role of TMEM16A( /-) for Ca(2+)-dependent Cl(-) secretion and provide the basis for novel treatments in cystic fibrosis, infectious diarrhea, and Sjoegren syndrome. PMID- 19679660 TI - N(alpha)-tosyl-L-phenylalanine chloromethyl ketone induces caspase-dependent apoptosis in transformed human B cell lines with transcriptional down-regulation of anti-apoptotic HS1-associated protein X-1. AB - N(alpha)-tosyl-L-phenylalanine chloromethylketone (TPCK) has been widely used to investigate signal transduction pathways that are involved in gene expression and cell survival/cell death. However, contradictory effects of TPCK on apoptosis have been reported, and the underlying signaling events leading to TPCK-induced promotion or prevention of apoptosis are not fully understood. Here, we show that TPCK induces caspase-dependent apoptosis in Epstein-Barr virus (EBV)-transformed human B cell lines with release of pro-apoptotic proteins from mitochondria. TPCK treatment also results in down-regulation of the anti-apoptotic proteins, cIAP1, cIAP2, and HAX-1, and caspase-dependent cleavage of the anti-apoptotic proteins, Bcl-2 and XIAP. Quantitative PCR analysis confirmed that the TPCK-induced down regulation of HAX-1 occurred at the transcriptional level, and experiments using the specific pharmacological inhibitor, Bay 11-7082, suggested that HAX-1 expression is subject to regulation by the transcription factor, NF-kappaB. B cell lines derived from patients with homozygous HAX1 mutations were more sensitive to TPCK-induced apoptosis when compared with normal donor cell lines. Furthermore, N-acetylcysteine effectively blocked TPCK-induced apoptosis in EBV transformed B cell lines and prevented the down-regulation or cleavage of anti apoptotic proteins. Taken together, our studies demonstrate that TPCK induces apoptosis in human B cell lines and exerts multiple effects on pro- and anti apoptotic factors. PMID- 19679662 TI - Identification of critical residues of the MyD88 death domain involved in the recruitment of downstream kinases. AB - MyD88 couples the activation of the Toll-like receptors and interleukin-1 receptor superfamily with intracellular signaling pathways. Upon ligand binding, activated receptors recruit MyD88 via its Toll-interleukin-1 receptor domain. MyD88 then allows the recruitment of the interleukin-1 receptor-associated kinases (IRAKs). We performed a site-directed mutagenesis of MyD88 residues, conserved in death domains of the homologous FADD and Pelle proteins, and analyzed the effect of the mutations on MyD88 signaling. Our studies revealed that mutation of residues 52 (MyD88(E52A)) and 58 (MyD88(Y58A)) impaired recruitment of both IRAK1 and IRAK4, whereas mutation of residue 95 (MyD88(K95A)) only affected IRAK4 recruitment. Since all MyD88 mutants were defective in signaling, recruitment of both IRAKs appeared necessary for activation of the pathway. Moreover, overexpression of a green fluorescent protein (GFP)-tagged mini-MyD88 protein (GFP-MyD88-(27-72)), comprising the Glu(52) and Tyr(58) residues, interfered with recruitment of both IRAK1 and IRAK4 by MyD88 and suppressed NF-kappaB activation by the interleukin-1 receptor but not by the MyD88-independent TLR3. GFP-MyD88-(27-72) exerted its effect by titrating IRAK1 and suppressing IRAK1-dependent NF-kappaB activation. These experiments identify novel residues of MyD88 that are crucially involved in the recruitment of IRAK1 and IRAK4 and in downstream propagation of MyD88 signaling. PMID- 19679663 TI - Clustering of neuronal K+-Cl- cotransporters in lipid rafts by tyrosine phosphorylation. AB - The neuronal K(+)-Cl(-) cotransporter (KCC2) is a membrane transport protein that extrudes Cl(-) from neurons and helps maintain low intracellular [Cl(-)] and hyperpolarizing GABAergic synaptic potentials. Depolarizing gamma-aminobutyric acid (GABA) responses in neonatal neurons and following various forms of neuronal injury are associated with reduced levels of KCC2 expression. Despite the importance for plasticity of inhibitory transmission, less is known about cellular mechanisms involved in more dynamic changes in KCC2 function. In this study, we investigated the role of tyrosine phosphorylation in KCC2 localization and function in hippocampal neurons and in cultured GT1-7 cells. Mutation to the putative tyrosine phosphorylation site within the long intracellular carboxyl terminus of KCC2(Y1087D) or application of the tyrosine kinase inhibitor genistein shifted the GABA reversal potential (E(GABA)) to more depolarized values, indicating reduced KCC2 function. This was associated with a change in the expression pattern of KCC2 from a punctate distribution to a more uniform distribution, suggesting that functional tyrosine-phosphorylated KCC2 forms clusters in restricted membrane domains. Sodium vanadate, a tyrosine phosphatase inhibitor, increased the proportion of KCC2 associated with lipid rafts membrane domains. Loss of tyrosine phosphorylation also reduced oligomerization of KCC2. A loss of the punctuate distribution and oligomerization of KCC2 and a more depolarized E(GABA) were seen when the 28-amino-acid carboxyl terminus of KCC2 was deleted. These results indicate that direct tyrosine phosphorylation of KCC2 results in membrane clusters and functional transport activity, suggesting a mechanism by which intracellular Cl(-) concentrations and GABA responses can be rapidly modulated. PMID- 19679665 TI - Phosphorylation of the yeast Rpb1 C-terminal domain at serines 2, 5, and 7. AB - The C-terminal domain (CTD) of Rpb1, the largest subunit of RNA polymerase II, acts as a binding platform for various mRNA processing and histone-modifying enzymes that act co-transcriptionally. These factors are targeted to specific phosphorylation states of the CTD that predominate at different stages of transcription. Within the repeating sequence YSPTSPS, serines 2 and 5 are major phosphorylation sites, but serine 7 phosphorylation was recently discovered in mammalian cells. Here we show that CTD serine 7 is also phosphorylated in yeast and that Ser-7(P) chromatin immunoprecipitation patterns resemble those of Ser 5(P). The basal factor TFIIH can phosphorylate Ser-7 in vitro and is necessary for Ser-7(P) in vivo. Interestingly, deletion of the CTD Ser-5(P) phosphatase Rtr1 leads to an increase in Ser-5(P) but not Ser-7(P). PMID- 19679664 TI - Adenovirus E1A inhibits SCF(Fbw7) ubiquitin ligase. AB - The SCF(Fbw7) ubiquitin ligase complex plays important roles in cell growth, survival, and differentiation via the ubiquitin-proteasome-mediated regulation of protein stability. Fbw7 (also known as Fbxw7, Sel-10, hCdc4, or hAgo), a substrate recognition subunit of SCF(Fbw7) ubiquitin ligase, facilitates the degradation of several proto-oncogene products by the proteasome. Given that mutations in Fbw7 are found in various types of human cancers, Fbw7 is considered to be a potent tumor suppressor. In the present study, we show that E1A, an oncogene product derived from adenovirus, interferes with the activity of the SCF(Fbw7) ubiquitin ligase. E1A interacted with SCF(Fbw7) and attenuated the ubiquitylation of its target proteins in vivo. Furthermore, using in vitro purified SCF(Fbw7) component proteins, we found that E1A directly bound to Roc1/Rbx1 and CUL1 and that E1A inhibited the ubiquitin ligase activity of the Roc1/Rbx1-CUL1 complex but not that of another RING-type ubiquitin ligase, Mdm2. Ectopically expressed E1A interacted with cellular endogenous Roc1/Rbx1 and CUL1 and decelerated the degradation of several protooncogene products that were degraded by SCF(Fbw7) ubiquitin ligase. Moreover, after wild-type adenovirus infection, adenovirus-derived E1A interacted with endogenous Roc1/Rbx1 and decelerated degradation of the endogenous target protein of SCF(Fbw7). These observations demonstrated that E1A perturbs protein turnover regulated by SCF(Fbw7) through the inhibition of SCF(Fbw7) ubiquitin ligase. Our findings may help to explain the mechanism whereby adenovirus infection induces unregulated proliferation. PMID- 19679667 TI - Peripheral vascular disease-related procedures in dialysis patients: predictors and prognosis. AB - BACKGROUND AND OBJECTIVES: Peripheral vascular disease (PVD) is prevalent among dialysis patients, and many dialysis patients undergo PVD-related procedures. We aimed to examine the risk factors for and prognosis after such procedures in the dialysis setting. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: In a national prospective cohort study of 1041 incident dialysis patients, we examined the factors that are associated with PVD procedures (lower extremity amputations and bypasses) after the start of dialysis. Adjusted risk for PVD procedures of various factors was estimated using multivariable Cox proportional hazards models. Incidence rates of subsequent cardiovascular events, infectious hospitalizations, PVD- and cardiovascular disease-related mortality, and all cause mortality were compared for those with and without a PVD procedure. RESULTS: Overall, 217 (21%) patients underwent a PVD procedure after the start of dialysis. For those without diabetes, only PVD history (relative hazard [RH] 2.9; 95% confidence interval [CI] 1.3 to 6.6) and increased fibrinogen (RH 1.2; 95% CI 1.0 to 1.5) predicted PVD procedures. For those with diabetes, increased serum phosphate (RH 1.2; 95% CI 1.1 to 1.4), along with decreased albumin, increased C reactive protein and fibrinogen, and lower SBP, was associated with risk for PVD procedures. Of those who had a procedure compared with those who did not, 68 versus 30% experienced a subsequent cardiovascular event, 85 versus 66% an infectious hospitalization, 11 versus 2% a PVD-related death, and 81 versus 59% all-cause death (mean follow-up 3.0 yr). CONCLUSIONS: Prognosis after PVD procedures is poor, and providers should be aware that risk factors for PVD procedures may differ by diabetes status. PMID- 19679666 TI - Cytomegalovirus promoter up-regulation is the major cause of increased protein levels of unstable reporter proteins after treatment of living cells with proteasome inhibitors. AB - Fluorescent unstable proteins obtained by the fusion of a fluorescent protein coding sequence with specific amino acid sequences that promote its fast degradation have become popular to gauge the activity of the ubiquitin/proteasome system in living cells. The steady-state levels of expression of these unstable proteins is low in agreement with their short half-lives, and they accumulate in the cell upon treatment with proteasome inhibitors. We show here that this accumulation is mainly due to transcriptional up-regulation of the cytomegalovirus promoter by proteasome inhibitors and mediated, at least in part, by AP1 transactivation. These simple facts put under quarantine conclusions reached about the activity of the ubiquitin/proteasome pathway in animal cells in culture or in transgenic mice, where popular cytomegalovirus-driven constructs are used, as transcriptional regulation of the expression of the reporter protein construct and not degradation of the unstable protein by the ubiquitin/proteasome system may contribute significantly to the interpretation of the results observed. PMID- 19679668 TI - Bacteremia associated with tunneled hemodialysis catheters: outcome after attempted salvage. AB - BACKGROUND AND OBJECTIVES: Treatment without catheter replacement (catheter salvage) has been described for bacteremia associated with tunneled venous catheters in hemodialysis patients, but few data are available on which to base an estimation of the likelihood of treatment success. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: In a prospective cohort study, all cases of catheter-associated bacteremia that occurred in a large dialysis center were identified during a 12-mo period. Catheter salvage was attempted according to a standard protocol in all cases in which a favorable early response to antibiotic therapy was seen, and patients were followed for at least 6 mo. Bacteremias, catheter changes, and all major clinical events were recorded. RESULTS: During a period covering 252,986 catheter days, 208 episodes were identified involving 133 patients, 74% of which were selected for attempted salvage. Salvage was successful in 66.1% of incident bacteremias with a very low complication risk (0.9%). Some bacteremias, however, recurred as late as 6 mo after the initial infection; salvage was less likely to be successful in treating recurrences. CONCLUSIONS: Appropriately used catheter salvage can be successful in approximately two thirds of cases; however, recurrences continue to occur up to 6 mo later and are unlikely to be cured without catheter replacement. PMID- 19679669 TI - Renal involvement in primary Sjogren's syndrome: a clinicopathologic study. AB - BACKGROUND & OBJECTIVES: Renal pathology and clinical outcomes in patients with primary Sjogren's syndrome (pSS) who underwent kidney biopsy (KB) because of renal impairment are reported. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Twenty-four of 7276 patients with pSS underwent KB over 40 years. Patient cases were reviewed by a renal pathologist, nephrologist, and rheumatologist. Presentation, laboratory findings, renal pathology, initial treatment, and therapeutic response were noted. RESULTS: Seventeen patients (17 of 24; 71%) had acute or chronic tubulointerstitial nephritis (TIN) as the primary lesion, with chronic TIN (11 of 17; 65%) the most common presentation. Two had cryoglobulinemic GN. Two had focal segmental glomerulosclerosis. Twenty patients (83%) were initially treated with corticosteroids. In addition, three received rituximab during follow-up. Sixteen were followed after biopsy for more than 12 mo (median 76 mo; range 17 to 192), and 14 of 16 maintained or improved renal function through follow-up. Of the seven patients presenting in stage IV chronic kidney disease, none progressed to stage V with treatment. CONCLUSIONS: This case series supports chronic TIN as the predominant KB finding in patients with renal involvement from pSS and illustrates diverse glomerular lesions. KB should be considered in the clinical evaluation of kidney dysfunction in pSS. Treatment with glucocorticoids or other immunosuppressive agents appears to slow progression of renal disease. Screening for renal involvement in pSS should include urinalysis, serum creatinine, and KB where indicated. KB with characteristic findings (TIN) should be considered as an additional supportive criterion to the classification criteria for pSS because it may affect management and renal outcome. PMID- 19679670 TI - Measuring patient survival on hemodialysis. PMID- 19679671 TI - Is calcitriol life-protective for patients with chronic kidney disease? PMID- 19679672 TI - DASH-style diet associates with reduced risk for kidney stones. AB - The impact of the Dietary Approaches to Stop Hypertension (DASH) diet on kidney stone formation is unknown. We prospectively examined the relation between a DASH style diet and incident kidney stones in the Health Professionals Follow-up Study (n = 45,821 men; 18 yr of follow-up), Nurses' Health Study I (n = 94,108 older women; 18 yr of follow-up), and Nurses' Health Study II (n = 101,837 younger women; 14 yr of follow-up). We constructed a DASH score based on eight components: high intake of fruits, vegetables, nuts and legumes, low-fat dairy products, and whole grains and low intake of sodium, sweetened beverages, and red and processed meats. We used Cox hazards regression to adjust for factors that included age, BMI, and fluid intake. Over a combined 50 yr of follow-up, we documented 5645 incident kidney stones. Participants with higher DASH scores had higher intakes of calcium, potassium, magnesium, oxalate, and vitamin C and had lower intakes of sodium. For participants in the highest compared with the lowest quintile of DASH score, the multivariate relative risks for kidney stones were 0.55 (95% CI, 0.46 to 0.65) for men, 0.58 (95% CI, 0.49 to 0.68) for older women, and 0.60 (95% CI, 0.52 to 0.70) for younger women. Higher DASH scores were associated with reduced risk even in participants with lower calcium intake. Exclusion of participants with hypertension did not change the results. In conclusion, consumption of a DASH-style diet is associated with a marked decrease in kidney stone risk. PMID- 19679673 TI - TbetaRI independently activates Smad- and CD2AP-dependent pathways in podocytes. AB - TGF-beta regulates differentiation, growth, and apoptosis of podocytes and mediates podocyte depletion in glomerulosclerosis. TGF-beta promotes proapoptotic signaling mediated by Smad3 but also activates prosurvival pathways such as phosphoinositide-3 kinase (PI3K)/AKT; the latter requires the CD2-associated adaptor protein (CD2AP) in podocytes. Whether the opposing activities mediated by Smad proteins and CD2AP involve molecular cross-talk is unknown. Here, we report that CD2AP-dependent early activation of the antiapoptotic PI3K/AKT pathway does not require TGF-beta receptor-regulated Smad2 and Smad3. We found that the C terminal region of CD2AP interacts directly with the cytoplasmic tail of the TGF beta receptor type I (TbetaRI) in a kinase-dependent manner and that the interaction between the TbetaRI and the p85 subunit of PI3K requires CD2AP. Consistent with the proapoptotic function of Smad signaling, Smad2/3-deficient podocytes were hyperproliferative and resistant to TGF-beta-induced growth inhibition and apoptosis. In contrast, CD2AP-deficient cells were hypoproliferative and hypersensitive to TGF-beta-induced apoptosis. In vivo, to determine the effects of reduced Smad3 or CD2AP gene dosage on podocyte apoptosis and proteinuria characteristic of TGF-beta1 transgenic mice, we generated TGF beta1 transgenic mice deficient for Smad3 or heterozygous for CD2AP. Smad3 deficiency ameliorated podocyte apoptosis, and CD2AP heterozygosity increased both podocyte apoptosis and proteinuria. These data define distinct canonical (Smad) and noncanonical (CD2AP/PI3K/AKT) pathways that arise from direct, independent interactions with the TbetaRI and that mediate opposing signals for podocyte death or survival. PMID- 19679674 TI - Hemoglobin vesicles, polyethylene glycol (PEG)ylated liposomes developed as a red blood cell substitute, do not induce the accelerated blood clearance phenomenon in mice. AB - The hemoglobin vesicle (HbV) is an artificial oxygen carrier encapsulating a concentrated hemoglobin solution in a liposome of which the surface is covered with polyethylene glycol (PEG). It was recently reported that repeated injections of PEGylated liposomes induce the accelerated blood clearance (ABC) phenomenon, in which serum anti-PEG IgM plays an essential role. To examine this issue, we investigated whether HbV induces the ABC phenomenon in mice at a dose of 0.1 mg Hb/kg, a dose that is generally known to induce the ABC phenomenon, or at 1400 mg Hb/kg, which is proposed for clinical use. At 7 days after the first injection of nonlabeled HbV (0.1 mg Hb/kg), the mice received HbV in which the Hb had been labeled with (125)I. After a second injection, HbV was rapidly cleared from the circulation, and uptake clearances in liver and spleen were significantly increased. In contrast, at a dose of 1400 mg Hb/kg, the pharmacokinetics of HbV was negligibly affected by repeated injection. It is interesting to note that IgM against HbV was produced 7 days postinjection at both of the above doses, and their recognition site was determined to be 1,2-distearoyl-sn-glycero-3 phosphatidylethanolamine-N-PEG in HbV. These results suggest that a clinical dose of HbV does not induce the ABC phenomenon, and that suppression of ABC phenomenon is caused by the saturation of phagocytic processing by the mononuclear phagocyte system. Thus, we conclude that induction of the ABC phenomenon would not be an issue in the dose regimen used in clinical settings. PMID- 19679675 TI - Effects of dose and route of administration on pharmacokinetics of (+ or -)-3,4 methylenedioxymethamphetamine in the rat. AB - Based on animal data, there is speculation that (+ or -)-3,4 methylenedioxymethamphetamine (MDMA) is neurotoxic to humans. Extrapolation of MDMA findings from animals to humans requires assessment of pharmacokinetics in various species, and low-dose administration data from rats are lacking. In this study, we examine MDMA pharmacokinetics in rats given low (2 mg/kg) and high (10 mg/kg) doses of racemic MDMA via intraperitoneal, subcutaneous, and oral routes. Repeated blood specimens were collected from venous catheters, and plasma was assayed for MDMA and its metabolites, 4-hydroxy-3-methoxymethamphetamine (HMMA) and 3,4-methylenedioxyamphetamine (MDA), by gas chromatography-mass spectrometry. After 2 mg/kg, maximum MDMA concentrations (C(max)) were approximately 200 ng/ml for intraperitoneal and subcutaneous routes, but less for the oral route. MDMA plasma half-lives were <1 h for low-dose groups, whereas HMMA and MDA half-lives were >2 h. After 10 mg/kg, MDMA areas under the curve (AUCs) were 21-fold (intraperitoneal), 10-fold (subcutaneous), and 36-fold (oral) greater than those at 2 mg/kg. In contrast, HMMA AUC values in high-dose groups were <3-fold above those at 2 mg/kg. Several new findings emerge from this report of low-dose MDMA pharmacokinetics in rats. First, 2 mg/kg MDMA in rats can produce MDMA C(max) values similar to those in humans, perhaps explaining why both species discriminate 1.5 mg/kg MDMA in laboratory paradigms. Second, our data provide additional support for nonlinear kinetics of MDMA in rats, and, analogous to humans, this phenomenon appears to involve impaired drug metabolism. Finally, given key similarities between MDMA pharmacokinetics in rats and humans, data from rats may be clinically relevant when appropriate dosing conditions are used. PMID- 19679678 TI - PopABC: a program to infer historical demographic parameters. AB - PopABC is a computer package for inferring the pattern of demographic divergence of closely related populations and species. The software performs coalescent simulation in the framework of approximate Bayesian computation (ABC). PopABC can also be used to perform Bayesian model choice to discriminate between different demographic scenarios. The program can be used either for research or for education and teaching purposes. AVAILABILITY AND IMPLEMENTATION: Source code and binaries are freely available at http://www.reading.ac.uk/ approximately sar05sal/software.htm. The program was implemented in C and can run on UNIX, MacOSX and Windows operating systems. PMID- 19679676 TI - Quantitative evaluation of the expression and activity of five major sulfotransferases (SULTs) in human tissues: the SULT "pie". AB - Expression levels of the major human sulfotransferases (SULTs) involved in xenobiotic detoxification in a range of human tissues (i.e., SULT "pies") are not available in a form allowing comparison between tissues and individuals. Here we have determined, by quantitative immunoblotting, expression levels for the five principal human SULTs-SULT1A1, SULT1A3/4, SULT1B1, SULT1E1, and SULT2A1-and determined the kinetic properties toward probe substrates, where available, for these enzymes in cytosol samples from a bank of adult human liver, small intestine, kidney, and lung. We produced new isoform-selective antibodies against SULT1B1 and SULT2A1, which were used alongside antibodies against SULT1A3 and SULT1A1 previously produced in our laboratory or available commercially (SULT1E1). Expression levels were derived using purified recombinant enzymes to construct standard curves for each individual isoform and immunoblot. Substantial intertissue and interindividual differences in expression were observed. SULT1A1 was the major enzyme (>50% of total, range 420-4900 ng/mg cytosol protein) in the liver, followed by SULT2A1, SULT1B1, and SULT1E1. SULT1A3 was completely absent from this tissue. In contrast, the small intestine contained the largest overall amount of SULT of any of the tissues, with SULT1B1 the major enzyme (36%), closely followed by SULT1A3 (31%), and SULT1A1, SULT1E1, and SULT2A1 more minor forms (19, 8, and 6% of total, respectively). The kidney and lung contained low levels of SULT. We provide a unique data set that will add value to the study of the role and contribution of sulfation to drug and xenobiotic metabolism in humans. PMID- 19679677 TI - Tissue distribution, ontogeny, and hormonal regulation of xenobiotic transporters in mouse kidneys. AB - Kidneys play important roles in the elimination of numerous endogenous and exogenous chemicals. In recent years, at least 37 xenobiotic transporters have been identified in mammalian kidneys. Although much progress has been made, information on 14 of these transporters (ATP-binding cassette [Abc] a1, apical sodium bile acid transporter [Asbt], breast cancer resistance protein, concentrative nucleoside transporter 1, equilibrative nucleoside transporter [Ent] 2, Ent3, sodium-phosphate cotransporter [Npt] 1, Npt2a, Npt2b, Npt2c, organic anion transporter [Oat] 5, organic anion-transporting polypeptide [Oatp] 4c1, peptide transporter 2, and uric acid transporter [Urat] 1) in kidneys is quite limited. Therefore, the purpose of the present study was to examine the tissue distribution, ontogeny, and hormonal regulation of these 14 transporters in kidneys of mice. Other than in kidneys, Npt2b is also highly expressed in liver and lung, Npt2c in liver and colon, Asbt in ileum, and Abca1 in liver, lung, testis, ovary, and placenta of mice. Most of these (13 of 14) transporters are lowly expressed in mouse kidneys until 15 days of age, which in part contributes to the immaturity of excretory function in fetal and newborn kidneys. One exception is Ent2, which is highly expressed before birth and gradually decreases after birth until reaching adult levels at 15 days of age. Gender divergent expression of male-predominant (Urat1 and Oatp4c1) and female predominant (Oat5) transporters in mouse kidneys is primarily due to stimulatory effects of androgens and estrogens, respectively. In conclusion, the mRNA expression of xenobiotic transporters in kidneys is determined by tissue, age, and sex hormones. PMID- 19679679 TI - Hypoxia-induced pulmonary hypertension: comparison of soluble epoxide hydrolase deletion vs. inhibition. AB - AIMS: The C-terminal domain of the soluble epoxide hydrolase (sEH) metabolizes epoxyeicosatrienoic acids (EETs) to their less active diols, while the N-terminal domain demonstrates lipid phosphatase activity. As EETs are potent vasoconstrictors in the pulmonary circulation, we assessed the development of pulmonary hypertension induced by exposure to hypoxia (10% O(2)) for 21 days in wild-type (WT) and sEH(-/-) mice and compared the effects with chronic (4 months) sEH inhibition. METHODS AND RESULTS: In isolated lungs from WT mice, acute hypoxic vasoconstriction (HPV) was potentiated by sEH inhibition and attenuated by an EET antagonist. After prolonged hypoxia, the acute HPV and sensitivity to the EET antagonist were increased, but potentiation of vasoconstriction following sEH inhibition was not evident. Chronic hypoxia also stimulated the muscularization of pulmonary arteries and decreased sEH expression in WT mice. In normoxic sEH(-/-) mice, acute HPV and small artery muscularization were greater than that in WT lungs and enhanced muscularization was accompanied with decreased voluntary exercise capacity. Acute HPV in sEH(-/-) mice was insensitive to sEH inhibition but inhibited by the EET antagonist and chronic hypoxia induced an exaggerated pulmonary vascular remodelling. In WT mice, chronic sEH inhibition increased serum EET levels but failed to affect acute HPV, right ventricle weight, pulmonary artery muscularization, or voluntary running distance. In human donor lungs, the sEH was expressed in the wall of pulmonary arteries, however, sEH expression was absent in samples from patients with pulmonary hypertension. CONCLUSION: These data suggest that a decrease in sEH expression is intimately linked to pathophysiology of hypoxia-induced pulmonary remodelling and hypertension. However, as sEH inhibitors do not promote the development of pulmonary hypertension it seems likely that the N-terminal lipid phosphatase may play a role in the development of this disease. PMID- 19679680 TI - Connexin 30 is expressed in the mouse sino-atrial node and modulates heart rate. AB - AIMS: This study aimed at characterizing expression and the functional role of the Gjb6 gene, encoding for connexin 30 (Cx30) protein, in the adult mouse heart. METHODS AND RESULTS: The expression of the Gjb6 gene in the mouse heart was investigated by RT-PCR and sequencing of amplified cDNA fragments. The sites of Gjb6 expression were identified in the adult heart using transgenic mice with reporter genes (Cx30(LacZ/LacZ) and Cx30(LacZ/LacZ)/Cx40(EGFP/EGFP) mice), as well as anti-HCN4 (hyperpolarization activated cyclic nucleotide-gated potassium channel 4) or anti-connexin antibodies. Cine-magnetic resonance imaging and telemetric ECG recordings were used to evaluate the impact of Cx30 deficiency on cardiac physiology. Gjb6 was shown to be expressed in the sinoatrial (SA) node of the adult mouse heart. Eighty from 100 nuclei on average were LacZ-positive in the SA node of Cx30(LacZ/LacZ) mice. No significant LacZ expression was seen in other cardiac tissues. Cx30 protein was identified in low abundance in the SA node of wild-type mice, as indicated by immunofluorescence experiments. Telemetric ECG recordings indicated that Cx30-deficient mice displayed a mean daily heart rate (HR) that was 9% faster than that measured in control mice (572 +/- 38 b.p.m. vs. 524 +/- 23, P < 0.05). This moderate tachycardia was still observed after inhibition of the autonomic nervous system, demonstrating that Cx30 deficiency resulted in changes in the intrinsic electrical properties of the SA node. Consistent with this hypothesis, Cx30(LacZ/LacZ) displayed a significant reduction of SDNN (standard deviation of the interbeat interval) compared with control mice. Increase of both the cardiac index (20%) and the end-diastolic volume to body weight ratio (16%) with no deficiency in ejection fraction or stroke volume were observed in mutant mice. An increase in cardiac index was interpreted as being a direct consequence of high HR, whereas large end-diastolic volume may be an indirect consequence of prolonged high HR. CONCLUSION: Cx30 is functionally expressed, in low abundance, in the SA node of the adult mouse heart where it participates in HR regulation. PMID- 19679681 TI - Protection of vascular cells from oxidative stress by proteasome inhibition depends on Nrf2. AB - AIMS: Increased levels of reactive oxygen species cause oxidative stress and severely damage lipids, proteins, and DNA. We have previously shown that partial proteasome inhibition induces an antioxidative gene pattern in endothelial cells. Here, we elucidate the mechanisms of proteasome inhibitor-mediated upregulation of antioxidative enzymes and cytoprotection. METHODS AND RESULTS: Non-toxic proteasome inhibition upregulated mRNA and protein expression of superoxide dismutase 1 (SOD1) and haem oxygenase 1 (HO1) in several human endothelial and vascular smooth muscle cell types. Transcriptional activation of these enzymes was shown by inhibition of RNA polymerase II and nuclear run-on assays. Transfection of endothelial cells with luciferase reporter constructs revealed that upregulation can be largely confined to an antioxidant response element (ARE), which proved to be sufficient for transcriptional activation of SOD1 and HO1. Co-transfection studies and bandshift analyses confirmed binding of the antioxidative transcription factor NF-E2-related factor 2 (Nrf2)-which was stabilized by proteasome inhibition as shown by immunoblots-to the ARE site of HO1. Experiments with aortic endothelial and smooth muscle cells from Nrf2 wild type and knockout mice revealed an essential role of Nrf2: in wild-type cells, proteasome inhibitor-mediated induction of SOD1 and HO1 was accompanied by protection of vascular cells against oxidative stress as determined by lactate dehydrogenase release assays. In contrast, proteasome inhibitor-mediated induction of antioxidative enzymes and cytoprotection were completely lost in cells from Nrf2 knockout mice. CONCLUSION: Nrf2-dependent transcriptional activation of antioxidative enzymes is crucial for proteasome inhibitor-mediated protection of vascular cells against oxidative stress. PMID- 19679682 TI - Quantitative detection of periodontopathic bacteria in atherosclerotic plaques from coronary arteries. AB - Oral pathogens, including periodontopathic bacteria, are thought to be aetiological factors in the development of cardiovascular disease. In this study, the presence of Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum periodonticum-simiae group, Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens and Tannerella forsythia in atheromatous plaques from coronary arteries was determined by real-time PCR. Forty-four patients displaying cardiovascular disease were submitted to periodontal examination and endarterectomy of coronary arteries. Approximately 60-100 mg atherosclerotic tissue was removed surgically and DNA was obtained. Quantitative detection of periodontopathic bacteria was performed using universal and species-specific TaqMan probe/primer sets. Total bacterial and periodontopathic bacterial DNA were found in 94.9 and 92.3 %, respectively, of the atheromatous plaques from periodontitis patients, and in 80.0 and 20.0 %, respectively, of atherosclerotic tissues from periodontally healthy subjects. All periodontal bacteria except for the F. nucleatum-periodonticum-simiae group were detected, and their DNA represented 47.3 % of the total bacterial DNA obtained from periodontitis patients. Porphyromonas gingivalis, A. actinomycetemcomitans and Prevotella intermedia were detected most often. The presence of two or more periodontal species could be observed in 64.1 % of the samples. In addition, even in samples in which a single periodontal species was detected, additional unidentified microbial DNA could be observed. The significant number of periodontopathic bacterial DNA species in atherosclerotic tissue samples from patients with periodontitis suggests that the presence of these micro-organisms in coronary lesions is not coincidental and that they may in fact contribute to the development of vascular diseases. PMID- 19679683 TI - Diversity of intestinal Escherichia coli populations in Nicaraguan children with and without diarrhoea. AB - Escherichia coli remains an important aetiological agent of infantile diarrhoea in Nicaragua. However, little is known about whether there is a high prevalence of endemic strains or whether infection is due to the epidemic spread of virulent clones. This study was undertaken to determine the diversity and distribution of clonal groups in a population of intestinal E. coli isolated from the faeces of children from Leon, Nicaragua, with (n=381) and without (n=145) diarrhoea, between March 2005 and September 2006. All samples had been screened previously for the presence of diarrhoeagenic E. coli (DEC) markers by multiplex PCR. From each sample, 8 E. coli colonies (where available) were analysed by biochemical fingerprinting (PhP-RE system), yielding a total of 4009 tested isolates. On average, three different biochemical phenotypes (BPTs) were found among the eight colonies analysed from each sample. The total diversity, measured as Simpson's diversity index (Di), was 0.97 among all 4009 isolates studied. Cluster analysis of data from all 4009 isolates revealed 24 common BPTs (identified in at least 1 % of the isolates) and 234 less common BPTs. Similar Di values were obtained among isolates from infants with and without diarrhoea, indicating that no widespread outbreak of DEC had occurred. Moreover, among samples that were positive for the DEC types enteroaggregative E. coli, enteropathogenic E. coli and enterotoxigenic E. coli (ETEC) carrying the eltB gene, the diversities were almost as high as among non-DEC samples, whereas samples positive for ETEC carrying estA, enteroinvasive E. coli and enterohaemorrhagic E. coli showed lower diversities, indicating the prevalence of virulent clonal groups among these samples. The PhenePlate patterns of the 24 common BPTs identified here were compared with those obtained from E. coli isolated in a cohort infant study performed in 1991-1992 in the same area. Only 4 % of the isolates from the 1990s were similar to any of the common BPTs found in the present study. PMID- 19679684 TI - Molecular characterization of a rare G1P[19] rotavirus strain from India: evidence of reassortment between human and porcine rotavirus strains. AB - This study pertains to the characterization of a human rotavirus strain (NIV929893) with a rare specificity of G1P[19]. Three structural genes (VP4, VP6 and VP7) and one non-structural gene (NSP4) of strain NIV929893 were subjected to RT-PCR for amplification of entire coding regions. All of the amplicons were sequenced to carry out phylogenetic analysis. The complete amino acid sequences of the VP7 and VP4 gene products showed clustering of the VP7 gene with G1 strains of human origin and the VP4 gene with P[19] strains of porcine origin. The two viral proteins VP6 and NSP4, described previously as genetically linked proteins, were shown to be subgroup II and genotype B of human and porcine origins, respectively. The findings of this study provide evidence of reassortment between VP7/VP6 genes of humans and VP4/NSP4 genes of porcine species and an independent segregation of VP6 and NSP4 genes in a group A human rotavirus strain with G1P[19] specificity. PMID- 19679687 TI - MICs and minimum fungicidal concentrations of amphotericin B, itraconazole, posaconazole and terbinafine in Sporothrix schenckii. AB - The in vitro susceptibility of 62 isolates of Sporothrix schenckii in its mycelial form, from Latin-American countries (Peru, Venezuela, Brazil and Uruguay) and Spain, to amphotericin B (AB), itraconazole (IZ), posaconazole (PZ) and terbinafine (TB) was determined by measuring the MICs and minimum fungicidal concentrations (MFCs) using a standardized Clinical and Laboratory Standards Institute method. In general, TB was the most active drug, with the lowest geometric mean (GM) MIC and MFC values amongst isolates from the five countries tested. IZ and PZ showed almost the same activity against all strains tested, except for isolates from Uruguay where IZ gave the highest GM MIC (10.68 mg l( 1)). AB showed the widest MIC range (0.03-16.0 mg l(-1)); however, this drug was less active against 79 % of isolates (MICs above 1 mg l(-1)). MFCs were 5 to 20 times higher than the MICs, but the lowest GM MFC and range values were found for TB. IZ and PZ gave the highest GM MFC. MFC may be a better predictor of therapeutic response than MIC, especially in immunosuppressed patients, making the use of IZ and PZ an inappropriate treatment. There were some differences in susceptibility according to the geographical source of the isolates, with the MIC being lower for TB in Venezuelan strains (P=0.066) and the MFC higher for PZ in Peruvian strains (P=0.02). Thus, geographical origin may be important for appropriate treatment, and may relate to the identification of species of the S. schenckii complex. PMID- 19679686 TI - Constitutive expression of the ileS-2 gene responsible for high-level mupirocin resistance in Staphylococcus aureus. AB - Mupirocin is a topical antimicrobial agent that is used for the treatment of skin and postoperative wound infections, and the prevention of nasal carriage of methicillin-resistant Staphylococcus aureus (MRSA). However, the prevalence of mupirocin resistance in S. aureus, particularly in MRSA, has increased. High level mupirocin resistance, with MICs >or=512 microg ml(-1), is mediated by the ileS-2 gene, which is located on conjugative plasmids. In the present study, we investigated whether mupirocin influences the expression of the ileS-2 gene responsible for high-level mupirocin resistance, and we present some evidence that this gene is not upregulated but constitutively expressed in S. aureus. PMID- 19679688 TI - Expert consensus on the management of erlotinib-associated cutaneous toxicity in the u.k. AB - Rash has been reported in up to 76% of patients with lung cancer who have received the epidermal growth factor receptor inhibitor (EGFRI) erlotinib. It has also been observed in patients treated with other agents that have a similar mode of action. Erlotinib-associated skin toxicity typically presents as a papulopustular, follicular, acneiform rash. In most cases, it is mild, transient, and well tolerated, but in 8%-12% of patients, it may be sufficiently severe and persistent to necessitate intervention. Increasingly strong data suggest that the incidence and severity of skin toxicity may be predictive of response and survival in patients treated with erlotinib. This has prompted some clinicians to consider "treatment to rash" (i.e., increasing the dosage until a rash appears) as a rational management strategy. In 2007, an international consensus was developed for the management of EGFRI-associated skin toxicity. Subsequently, a multidisciplinary group (the U.K. Erlotinib Skin Toxicity Management Consensus Group) met to validate and modify the international recommendations for U.K. use, with specific reference to erlotinib. Although many aspects of the international consensus were approved by the group as being relevant for the U.K., certain parts were modified. The resulting expert opinion is a practical and workable version of the international proposal that considers all applicable national issues regarding the management of erlotinib-associated skin toxicity. PMID- 19679685 TI - An imprint method for detecting leptospires in the hamster model of vaccine mediated immunity for leptospirosis. AB - In determining the efficacy of new vaccine candidates for leptospirosis, the primary end point is death and an important secondary end point is sterilizing immunity. However, evaluation of this end point is often hampered by the time consuming demands and complexity of methods such as culture isolation (CI). In this study, we evaluated the use of an imprint (or touch preparation) method (IM) in detecting the presence of leptospires in tissues of hamsters infected with Leptospira interrogans serovar Copenhageni. In a dissemination study, compared to CI, the IM led to equal or improved detection of leptospires in kidney, liver, lung and blood samples collected post-infection and overall concordance was good (kappa=0.61). Furthermore, in an evaluation of hamsters immunized with a recombinant leptospiral protein-based vaccine candidate and subsequently challenged, the agreement between the CI and IM was very good (kappa=0.84). These findings indicate that the IM is a rapid method for the direct observation of Leptospira spp. that can be readily applied to evaluating infection in experimental animals and determining sterilizing immunity when screening potential vaccine candidates. PMID- 19679689 TI - The future burden of cancer in London compared with England. AB - BACKGROUND: The future burden of cancer in England is predicted to increase by 33% by 2020. Those planning health services are often interested in predictions at a local level. This study aimed to estimate the future burden of cancer in London and compare this with estimates for England. METHODS: Predictions for London were based on cancer registration data (1985-2003) and population projections up until 2024. The number of future cases and age-standardized incidence rates (ASRs) were projected using an age-period-cohort model developed for the analysis of cancer trends and projections in the Nordic countries. Estimates for England were taken from an earlier publication. RESULTS: In London, ASRs for all cancers (excluding non-melanoma skin cancer) decreased for both sexes, whereas in England, ASRs decreased in males and increased slightly in females. In London, the number of cases for both sexes is predicted to increase from approximately 27,000 in 2002 to approximately 28,500 in 2022, an increase of 5%. In England, a greater increase is predicted, from approximately 224,000 in 2001 to approximately 299,000 in 2020, an increase of 33%. CONCLUSIONS: Projections of the future burden of cancer are particularly sensitive to demographic population trends. They are likely to be different for cities compared with rural areas or entire countries. PMID- 19679690 TI - Review of preoperative magnetic resonance imaging (MRI) in breast cancer: should MRI be performed on all women with newly diagnosed, early stage breast cancer? AB - Randomized controlled trials have shown equivalent survival for women with early stage breast cancer who are treated with breast-conservation therapy (local excision and radiotherapy) or mastectomy. Decades of experience have demonstrated that breast-conservation therapy provides excellent local control based on defined standards of care. Magnetic resonance imaging (MRI) has been introduced in preoperative staging of the affected breast in women with newly diagnosed breast cancer because it detects additional foci of cancer that are occult on conventional imaging. The median incremental (additional) detection for MRI has been estimated as 16% in meta-analysis. In the absence of consensus on the role of preoperative MRI, we review data on its detection capability and its impact on treatment. We outline that the assumptions behind the adoption of MRI, namely that it will improve surgical planning and will lead to a reduction in re excision surgery and in local recurrences, have not been substantiated by trials. Evidence consistently shows that MRI changes surgical management, usually from breast conservation to more radical surgery; however, there is no evidence that it improves surgical care or prognosis. Emerging data indicate that MRI does not reduce re-excision rates and that it causes false positives in terms of detection and unnecessary surgery; overall there is little high-quality evidence at present to support the routine use of preoperative MRI. Randomized controlled trials are needed to establish the clinical, psychosocial, and long-term effects of MRI and to show a related change in treatment from standard care in women newly affected by breast cancer. PMID- 19679691 TI - Impaired IL-10-dependent induction of tolerogenic dendritic cells by CD4+CD25hiCD127lo/- natural regulatory T cells in human allergic asthma. AB - RATIONALE: Tolerogenic dendritic cells and natural regulatory T cells have been implicated in the process of infectious tolerance in human allergic asthma. However, the significance of the influence of natural regulatory T cells on tolerogenic dendritic cells in the disease has not been investigated. OBJECTIVES: We aimed to characterize the mechanism of induction of the tolerogenic phenotype in circulating blood dendritic cells by allergic asthmatic natural regulatory T cells. METHODS: The study was performed in a cohort of 21 subjects with allergic asthma, 21 healthy control subjects, and 21 subjects with nonallergic asthma. We cultured blood dendritic cells with natural regulatory T cells to study the induction of tolerogenic dendritic cells. Flow cytometry and proliferation assays were employed to analyze phenotype and function of dendritic cells as well as IL 10 production from natural regulatory T cells. MEASUREMENTS AND MAIN RESULTS: Dendritic cells cultured with natural regulatory T cells up-regulated IL-10, down regulated costimulatory molecules, and stimulated the proliferation of CD4(+)CD25(-) effector T cells less potently. Allergic asthmatic natural regulatory T cells were significantly less efficient in inducing this tolerogenic phenotype of dendritic cells compared with healthy control and nonallergic asthmatic counterparts. Furthermore, this defective function of natural regulatory T cells was associated with their decreased IL-10 expression, disease severity, and could be reversed by oral corticosteroid therapy. CONCLUSIONS: These results provided the first evidences of impaired induction of tolerogenic dendritic cells mediated by natural regulatory T cells in human allergic asthma. PMID- 19679692 TI - The triterpenoid CDDO-imidazolide confers potent protection against hyperoxic acute lung injury in mice. AB - RATIONALE: Oxygen supplementation (e.g., hyperoxia) is used to support critically ill patients with noninfectious and infectious acute lung injury (ALI); however, hyperoxia exposure can potentially further contribute to and/or perpetuate preexisting ALI. Thus, developing novel therapeutic agents to minimize the side effects of hyperoxia is essential to improve the health of patients with severe ALI and respiratory dysfunction. We have previously shown that mice with a genetic disruption of the Nrf2 transcription factor, which squelches cellular stress by up-regulating the induction of several antioxidant enzymes and proteins, have greater susceptibility to hyperoxic lung injury. Moreover, we have recently demonstrated that Nrf2-deficiency impairs the resolution of lung injury and inflammation after nonlethal hyperoxia exposure. OBJECTIVES: To test the hypothesis that amplification of endogenous Nrf2 activity would prevent or dampen ALI induced by hyperoxia. METHODS: Here, we tested our hypothesis using a synthetic triterpenoid compound CDDO-imidazole (CDDO-Im) (1-[2-cyano-3-,12 dioxooleana-1,9(11)-dien-28-oyl] imidazole) in Nrf2-sufficient and Nrf2-deficient mice subjected to hyperoxia-induced ALI. MEASUREMENTS AND MAIN RESULTS: We demonstrate that oral administration of CDDO-Im at a dose of 30 micromol/kg body weight during the hyperoxic exposure is sufficient to markedly attenuate hyperoxia-induced ALI in Nrf2-sufficient but not Nrf2-deficient mice. This protection by the CDDO-Im against hyperoxic insult was accompanied by increased levels of Nrf2-regulated cytoprotective gene expression and reduced levels of DNA damage in the lung. CONCLUSIONS: These results suggest that up-regulation of Nrf2 signaling by CDDO-Im or its analogs may provide a novel therapeutic strategy to minimize the adverse effects of hyperoxia. PMID- 19679693 TI - Borderline pulmonary arterial pressure is associated with decreased exercise capacity in scleroderma. AB - RATIONALE: Pulmonary arterial hypertension is associated with impaired exercise capacity and decreased survival in patients with scleroderma. Randomized controlled studies showed significant benefit of targeted therapies in patients with a resting mean pulmonary arterial pressure (MPAP) greater than 25 mm Hg. The clinical relevance of pulmonary arterial pressure values in the upper normal range is unknown. OBJECTIVES: To examine the clinical relevance of pulmonary arterial pressure in scleroderma patients. METHODS: After a noninvasive screening program, 29 patients with systemic sclerosis without significant lung fibrosis and without known pulmonary arterial hypertension underwent right heart catheterization and simultaneous cardiopulmonary exercise test. A six-minute walk distance (6MWD) was determined within 48 hours. MEASUREMENTS AND MAIN RESULTS: A resting MPAP above the median (17 mm Hg) was associated with decreased 6MWD (396 +/- 71 vs. 488 +/- 76 m; P < 0.005) and peak Vo(2) (76 +/- 11% vs. 90 +/- 24%; P = 0.05). Resting pulmonary vascular resistance was inversely correlated with 6MWD (r = 0.45; P < 0.05). At 25 and 50W, MPAP above the median (23 and 28 mm Hg) was associated with decreased 6MWD (P < 0.005; P < 0.0005). At peak exercise, MPAP showed no association with 6MWD or peak Vo(2); however, cardiac index was positively (r = 0.45; P < 0.05) and pulmonary vascular resistance was negatively correlated with 6MWD (r = -0.38; P < 0.05). CONCLUSIONS: MPAP and resistance in the upper normal range at rest and moderate exercise are associated with decreased exercise capacity and may indicate early pulmonary vasculopathy in patients with systemic sclerosis. Investigations on the prognostic and therapeutic implications of such borderline findings are warranted. Clinical trial registered with http://www.clinicaltrials.gov (NCT00609349). PMID- 19679695 TI - HSC mobilization: new incites and insights. PMID- 19679694 TI - Increasing proportions of advanced pulmonary tuberculosis reported in the United States: are delays in diagnosis on the rise? AB - RATIONALE: Delays in the diagnosis of tuberculosis (TB) can result in progression to advanced disease. Patients with pulmonary TB and advanced disease are more likely to transmit disease and fail treatment. OBJECTIVES: To examine clinical, epidemiological, and geographic factors associated with advanced pulmonary TB to further understanding of delayed diagnosis and transmission. METHODS: Pulmonary tuberculosis cases in persons older than 15 years of age reported to the U.S. National Tuberculosis Surveillance System with advanced disease (cavitation on chest radiograph and acid-fast bacilli smear-positive sputum result) were compared with those without advanced disease using trend and binomial regression analysis. MEASUREMENTS AND MAIN RESULTS: There were 35,584 cases of advanced pulmonary tuberculosis (APT) and 125,077 cases of non-APT reported from 1993 through 2006. Proportions of pulmonary TB cases with APT increased from 18.5% in 1993 to 26.1% in 2006, and the increase in the proportion of APT was most notable for national TB rates below 6.6 per 100,000. At the county level, the association between APT and low TB incidence has grown incrementally since 2000. The proportion of APT increased greatest among whites (65.4%), the employed (63.3%), and the U.S. born (59.2%). The prevalence of APT was 44% greater among persons with multidrug-resistant TB compared with those without it. CONCLUSIONS: This study highlights the need for TB diagnosis at early stages of the disease to minimize APT and decrease the risk of transmission. Additional efforts should concentrate on reducing time to treatment initiation in low-incidence areas and among groups traditionally seen as being at low risk for TB disease. PMID- 19679696 TI - Stem cell transplantation in AIDS-lymphoma. PMID- 19679697 TI - Better positioned in stem cells. PMID- 19679698 TI - Platelet genomics beats the catch-22. PMID- 19679699 TI - TRECing long-term success in SCID. PMID- 19679700 TI - Long-term maintenance of retest learning in young old and oldest old adults. AB - This study examined the maintenance of retest learning benefits in young old and oldest old adults over an 8-month period in 3 cognitive abilities: reasoning, perceptual-motor speed, and visual attention. Twenty-four young old (aged 70-79 years, M = 74.2) and 23 oldest old adults (aged 80-90 years, M = 83.6) who participated in a previously published study (Yang, L., Krampe, R. T., & Baltes, P. B. [2006]. Basic forms of cognitive plasticity extended into the oldest-old: Retest learning, age, and cognitive functioning. Psychology and Aging, 21, 372 378) returned after an 8-month delay to complete 2 follow-up retest sessions. The results demonstrated that both young old and oldest old groups maintained about 50% of the original retest learning benefits. This extends the earlier findings of substantial long-term cognitive training maintenance in young old adults to a context of retest learning with oldest old adults, and thus portrays a positive message for cognitive plasticity of the oldest old. PMID- 19679701 TI - Women and their partners' perceptions of the key roles of the labor and delivery nurse. AB - This descriptive qualitative study examined the perspectives of women and their partners regarding the key roles of the labor and delivery nurse during labor and birth.Ten couples were interviewed separately.The data analysis, conducted through independent and team analysis, was both iterative and interpretive. Participants identified four key roles of the labor and delivery nurse: support person, educator, patient advocate, and provider of continuity. Nurses provided both physical and emotional support.As an educator, they normalized the birth experience and served as a coach for the couple. Nurses advocated on behalf of the woman in labor, particularly when there was an adverse event. The continuity of care provided by the nurses wove the above roles into a cohesive whole. Findings provide important information for nursing educators, supervisors, and hospital administrators to reinforce the meaningful roles nurses serve in the labor and birth experiences of women and their partners. PMID- 19679702 TI - Job stressors and long-term sick leave due to depressive disorders among Japanese male employees: findings from the Japan Work Stress and Health Cohort study. AB - BACKGROUND: Research on the association between job strain or other job stressors and depressive disorders is still limited. The purpose of the present study was to investigate the prospective association of job strain, role stressors and job insecurity with long-term sick leave due to depressive disorders. METHODS: A prospective study was conducted of a total of 15 256 men aged 18-67 years with no previous history of mental disorders employed in six manufacturing factories located in several regions of Japan. At baseline, they were surveyed using a self administered questionnaire, including self-reported measures of job strain, as well as its components (job overload and job control), role stressors (role ambiguity and role conflict), social support at work, job insecurity and other demographic and psychological covariates. During the follow-up, a long-term sick leave of 30 days or more due to depressive disorders was recorded. RESULTS: During 5.14 years of follow-up on average, 47 incident cases of sick leave of 30 days or more due to depressive disorders were observed. High job control at baseline was associated with a lower risk of long-term sick leave due to depressive disorders, after adjusting for demographic variables, depressive symptoms and neuroticism at baseline (hazard ratio 0.28, 95% CI 0.11 to 0.71); high role ambiguity was associated with the higher risk (hazard ratio 3.49, 95% CI 1.43 to 8.49). CONCLUSION: Job control and role ambiguity may be important predictors of long-term sick leave due to depressive disorders among male employees, independent of depressive symptoms and neuroticism. PMID- 19679703 TI - Psychopathology among children and adolescents in child welfare: a comparison across different types of placement in Flanders, Belgium. AB - BACKGROUND: Children and adolescents in child welfare are characterised by a high rate of psychopathology. However, prevalence estimates vary greatly, and comparisons between studies are limited owing to different target populations, measurement tools and how results are presented. In addition, little is known about the situation of children under child welfare care still living with their parents. METHODS: The Achenbach System of Empirically Based Assessment and the Strengths and Difficulties Questionnaire used to measure psychopathology, were administered to parent, agency carer and adolescent (if older than 11). Data of 292 children aged 3-17 years (mean age, 12.13 years) with at least one respondent of four types of placement, namely foster care, residential care, day care and home-based care, were included (response rate, 78%). Socioeconomic characteristics and psychopathology were examined across these four types of placement. RESULTS: Overall, the proportion of children scoring within the clinical range according to at least one informant was 56% according to the Strengths and Difficulties Questionnaire and 54% according to the Achenbach System of Empirically Based Assessment. Rates were highest among children living at home and significantly lower among children in out-of-home placement, even after correcting for age, income and duration of the ongoing placement. Adolescents scored themselves lower on psychopathology in comparison with their parents and the agency carer. CONCLUSIONS: The prevalence of psychopathology in this child welfare population was very high; however, the findings were consistent with results obtained in previous studies. The prevalence estimates differed depending on the type of placement: the highest rates were found among children in home-based care, and children in foster care suffered less psychopathology. The findings stress the vulnerable mental status of children in child welfare and the need for additional support for child welfare professionals and children, as well as their parents, especially for those living with their parents. PMID- 19679704 TI - Uptake of treatment for osteoporosis and compliance after bone density measurement in the community. AB - BACKGROUND: Management of osteoporosis is imperfect because patients may not start, persist or comply with treatment. This study was aimed to identify baseline variables associated with women failing to start, persist or comply with bisphosphonate treatment. METHODS: 254 women >50 years old were selected 5 years after having a bone densitometry (bone mineral density (BMD)) diagnosis of osteoporosis. At the outset, information about osteoporosis was sent to the general practitioner (GP). Women were not under pressure at the outset to start or comply, and they and their GP were unaware that follow-up studies would take place. Patient survival was identified from the National Health Service Strategic Tracing Service, prescription data from GP records and baseline data from the initial questionnaire. Persistence was defined as at least one prescription issued per year and compliance as having a medicine possession ratio of >or=80% for each of 5 years. RESULTS: 38% failed to start treatment. Failure was associated with higher BMD Z score and residence in a nursing/residential home. Half of those starting and alive at 5 years persisted with treatment, whereas only 23% achieved a medicine possession ratio of >or=80%. Persistence was associated with a lower comorbidity index and compliance with a lower BMD Z score and a fall before starting treatment. CONCLUSIONS: Treatment was low, especially in nursing/residential homes where known low treatment prevalence appears to be associated with non-initiation. The degree of depression of BMD (not just low BMD) was associated with better initiation and compliance. The association of falls with compliance suggests that fall clinics may be able to play a part in improving osteoporosis management. PMID- 19679705 TI - Association of traffic-related air pollution with cognitive development in children. AB - BACKGROUND: Air pollution from traffic has been associated with cardiorespiratory diseases in children and adults, but there is little information on its potential neurotoxic effects. This study aimed to investigate the association between exposure to nitrogen dioxide (NO(2)), as a marker of traffic-related air pollution, and cognitive development in children. METHODS: A population-based birth cohort from southern Spain was followed from the age of 4 years for 1 year. Complete data for analyses were gathered on 210 children living in urban and rural areas. NO(2) exposure was predicted by means of land use regression models. A standardised version of the McCarthy Scales of Children's Abilities (MSCA) was used to assess children's motor and cognitive abilities. Multivariate analyses were performed to evaluate the relation between exposure to NO(2) and MSCA outcomes, adjusting for potential confounders. RESULTS: A negative effect of NO(2) was found across all MSCA subscales, despite low predicted NO(2) exposure levels (5-36 microg/m(3)). Children exposed to higher NO(2) (>24.75 microg/m(3)) showed a decrease of 4.19 points in the general cognitive score and decreases of 6.71, 7.37 and 8.61 points in quantitative, working memory and gross motor areas, respectively. However, except for gross motor function, associations were not statistically significant. CONCLUSION: Although results were not statistically significant, the associations found between exposure to NO(2) and cognitive functions suggest that traffic-related air pollution may have an adverse effect on neurodevelopment, especially early in life, even at low exposure levels. PMID- 19679706 TI - All-cause and diagnosis-specific sickness absence as a predictor of sustained suboptimal health: a 14-year follow-up in the GAZEL cohort. AB - background: Previous studies show that sickness absence predicts health, but it is unclear whether this association is persistent over time and whether specific diseases underlie long-term associations. The aim of this study was to investigate overall and diagnosis-specific sickness absences as predictors of sustained suboptimal health. METHODS: Prospective occupational cohort study of 15 320 employees (73% men) aged 37-51. Sickness absence records in 1990-1992, including 13 diagnostic categories, were examined in relation to self-rated health measured annually for the years 1993-2006. RESULTS: 3385 employees (22%) had >30 days of sickness absence and 5564 (36%) 1-30 days during the 3-year exposure window. Repeated-measures logistic regression analysis adjusted for age, sex, occupational status and chronic diseases show that employees with >30 absence days, compared with those with no absences, had 2.14 (95% CI 2.00 to 2.29) times higher odds for suboptimal health over the 14 years of follow-up. Retirement did not dilute this association. Nine sickness absence diagnostic categories, such as diseases of the nervous, circulatory, metabolic, musculoskeletal, sensory and gastrointestinal systems, cancer, mental disorders and external causes, independently predicted increased risk of sustained suboptimal health. CONCLUSIONS: There is a remarkably persistent association between sickness absence and future long-term self-rated health status for the majority of diagnostic categories for sickness absence. This suggests that the association between sickness absence and health is ubiquitous and not driven by a limited number of rare and severe diseases. PMID- 19679707 TI - Suicide in young Norwegians in a life course perspective: population-based cohort study. AB - BACKGROUND: Suicide is a leading cause of death in young adults. Several risk factors are well known, especially those related to adult mental health. However, some risk factors may have their origin in the very beginning of life. This study examines suicide in the general Norwegian population in a life course perspective, with a main focus on early life factors. METHODS: In this study, several national registers were linked, supplying personal data on biological and social variables from childhood to young adult age. Participants were all Norwegians live born during the period 1967-1976, followed up through 2004. Persons who died or emigrated before the year of their 19th birthday, at which age follow-up started, were excluded. Thus, the study population comprised 610 359 persons, and the study outcome was completed suicide. RESULTS: 1406 suicides (0.23%) were recorded, the risk being four times higher in men than in women. Suicide risk factors included not being firstborn (adjusted HR in men and women (95% CIs): 1.19 (1.05 to 1.36) and 1.42 (1.08 to 1.88)), instability of maternal marital status during childhood, parental suicide (mainly in women), low body mass index (only investigated in men), low education and indications of severe mental illness. CONCLUSIONS: Suicide in young adults may be rooted in early childhood, and the effect is likely to act through several mechanisms, some of which may be linked to the composition and stability of the parental home. A life course perspective may add to our understanding of suicide. PMID- 19679708 TI - Association of diarrhoea, poor hygiene and poor social conditions in childhood with blood pressure in adulthood. AB - BACKGROUND: Previous research has suggested that dehydration in infancy may lead to high blood pressure in later life because of sodium retention. The purpose of this study was to examine the effect of poor hygiene of the child, poor social and poor housing conditions at home and diarrhoea in childhood as proxies for dehydration on high blood pressure in later life. METHODS: Data were from a subset of participants in the Kuopio Ischaemic Heart Disease Risk Factor Study, a population-based cohort study in eastern Finland. Information on childhood factors was collected from school health records (n=952), from the 1930s to the 1950s. Adult data were obtained from baseline examinations of the Kuopio Ischaemic Heart Disease Risk Factor Study cohort (n=2682) in 1984-1989. RESULTS: Men who had poor hygiene in childhood had on average 4.07 mm Hg (95% CI 0.53 to 7.61) higher systolic blood pressure than men who had good or satisfactory hygiene in childhood in the age-adjusted analysis. Reports of diarrhoea were not associated with adult blood pressure. CONCLUSIONS: The authors' findings suggest that poor hygiene and living in poor social conditions in childhood are associated with higher systolic blood pressure in adulthood. Reported childhood diarrhoea did not explain the link between hygiene and high blood pressure in adulthood. PMID- 19679709 TI - Socioeconomic differences in second-hand smoke exposure among children in Scotland after introduction of the smoke-free legislation. AB - BACKGROUND: To examine the impact of the Scottish smoke-free legislation on social inequalities in secondhand smoke (SHS) exposure among primary school children. METHODS: Comparison of nationally representative, cross-sectional, class-based surveys carried out in the same schools before and after legislation. Participants were 2532 primary school children (primary 7; aged around 11 y) surveyed in January 2006 (before legislation) and 2389 in January 2007 (after legislation). Outcome measures were salivary cotinine concentrations, self reported family socioeconomic classification (family SEC) and family affluence scale (FAS). RESULTS: After adjusting for number of smoking parents, mean cotinine concentration varied significantly across both family SEC and FAS groups, and increased significantly stepwise from high to low family SEC/FAS. Mean cotinine fell in all family SEC/FAS groups after legislation. The relative drop in mean cotinine was equal across all family SEC/FAS groups. Adding an interaction term between survey-year and family SEC/FAS to the model showed an increase in inequalities over time, but was only significant at the 93% level using FAS and 73% using family SEC. CONCLUSION: Inequalities in SHS exposure exist among 11-year-old children in Scotland. Smoke-free legislation has reduced exposure to SHS among all children. Although the greatest absolute reduction in cotinine is observed in the lowest SEC/FAS group, cotinine levels remain highest for this group and there is a suggestion of possible increases in inequalities, which may warrant longer-term monitoring. PMID- 19679710 TI - Socioeconomic status and the risk of major depression: the Canadian National Population Health Survey. AB - BACKGROUND: There are few longitudinal studies investigating the risk of major depression by socioeconomic status (SES). In this study, data from the longitudinal cohort of Canadian National Population Health Survey were used to estimate the risk of major depressive episode (MDE) over 6 years by SES levels. METHODS: The National Population Health Survey used a nationally representative sample of the Canadian general population. In this analysis, participants (n=9589) were followed from 2000/2001 (baseline) to 2006/2007. MDE was assessed using the Composite International Diagnostic Interview--Short Form for Major Depression. RESULTS: Low education level (OR=1.86, 95% CI 1.28 to 2.69) and financial strain (OR=1.65, 95% CI 1.19 to 2.28) were associated with an increased risk of MDE in participants who worked in the past 12 months. In those who did not work in the past 12 months, participants with low education were at a lower risk of MDE (OR=0.43, 95% CI 0.25 to 0.76), compared with those with high education. Financial strain was not associated with MDE in participants who did not work. Working men who reported low household income (12.9%) and participants who did not work and reported low personal income (5.4%) had a higher incidence of MDE than others. CONCLUSIONS: SES inequalities in the risk of MDE exist in the general population. However, the inequalities may depend on measures of SES, sex and employment status. These should be considered in interventions of reducing inequalities in MDE. MDE history is an important factor in studies examining inequalities in MDE. PMID- 19679711 TI - Socioeconomic variations in the prevalence and incidence of Parkinson's disease: a population-based analysis. AB - BACKGROUND: For many chronic conditions, lower socioeconomic status is associated with higher rates of disease. Previous research has not investigated whether this inverse relationship exists for Parkinson's disease (PD). The purpose was to investigate the association between socioeconomic status and prevalence and incidence of PD. METHODS: The study was conducted using population-based administrative data from Manitoba, Canada. PD cases were identified from diagnoses in hospital and physician records. Area-level census data on average household income and residential postal codes in health insurance registration files were used to assign PD cases to urban and rural income quintiles. Annual adjusted prevalence and incidence estimates were calculated for fiscal years 1987/88-2006/07. Hypotheses about differences between quintiles in average estimates and average rates of change were tested using generalised linear models with generalised estimating equations. RESULTS: The estimated prevalence of PD increased over the 20-year-period but incidence remained unchanged. In urban regions, average prevalence and incidence estimates were significantly higher for the lowest income quintile than the highest quintile. In rural regions, average prevalence estimates were significantly higher for the lowest quintile than for the highest quintile but incidence estimates were not significantly different. The annual rate of increase in the PD prevalence was significantly different for the lowest urban and rural income quintiles. CONCLUSIONS: There is a greater burden of PD in low-income areas. An understanding of socioeconomic inequalities is useful when formulating hypotheses about factors associated with disease onset and developing equity-oriented policies about access to healthcare resources. PMID- 19679712 TI - Does organisational justice protect from sickness absence following a major life event? A Finnish public sector study. AB - BACKGROUND: It has been shown that fairness perceptions have a strong impact on health, especially under conditions of great work stress. The aim of this study was to extend previous research in studying whether working in high justice workplace would protect from health effects following environmental stressors outside work. METHODS: Using a prospective longitudinal design, the relationships between organisational justice and sickness-related absences both before and after a major life event among 25 459 public sector employees working in 2551 work units were studied. Sickness absences covered the period from 36 months before the event until 30 months after the event. RESULTS: The increase in sickness absences after the event was larger and stayed at a higher level even 30 months after the event, among those who perceived the management practices in their work unit to be relatively unfair. Similar patterns were found for each of the distributive, procedural and interactional dimensions of organisational justice. CONCLUSIONS: Fair organisational and managerial procedures may buffer the negative health effects of psychosocial health risks outside work. PMID- 19679713 TI - The European Perinatal Health Report: comparing the health and care of pregnant women and newborn babies in Europe. PMID- 19679714 TI - Imprint regulatory elements as epigenetic biosensors of exposure in epidemiological studies. PMID- 19679715 TI - The effect of neighbourhood income and deprivation on pregnancy outcomes in Amsterdam, The Netherlands. AB - BACKGROUND: Studies suggest that the neighbourhoods in which people live influence their health. The main objective of this study was to investigate the associations of neighbourhood-level income and unemployment/social security benefit on pregnancy outcomes: preterm delivery, small for gestational age (SGA), pregnancy-induced hypertension (PIH) and miscarriage/perinatal death in Amsterdam. METHODS: A random sample of 7883 from 82 neighbourhoods in Amsterdam. Individual-level data from the Amsterdam Born Children and their Development (ABCD) study were linked to data on neighbourhood-level factors. Multilevel logistic regression was used to estimate odds ratios and neighbourhood-level variance. RESULTS: After adjustment for individual-level factors, women living in low-income neighbourhoods (third, second and first quartiles) were more likely than women living in high-income neighbourhoods (fourth quartile) to have SGA births: OR 1.32 (95% CI 1.04 to 1.68), 1.42 (1.11 to 1.82) and 1.62 (1.25 to 2.08) respectively. Women living in the quartile of neighbourhoods with the highest unemployment/social security benefit were more likely than those living in the quartile with the lowest unemployment/social security benefit to have SGA births 1.36 (1.08 to 1.72). The neighbourhood-level variance was significant only for SGA births. No significant associations were found between neighbourhood level factors and other pregnancy outcomes. CONCLUSION: The findings suggest that neighbourhood income and deprivation are related to SGA births. More research is needed to explore possible mechanisms underlying poor neighbourhood environment and pregnancy outcomes, in particular through stress mechanisms. Such information might be necessary to help improve maternal and fetal health. PMID- 19679717 TI - A plea for basic science. PMID- 19679716 TI - First-time primary caregivers' experience of caring for young adults with first episode psychosis. AB - Becoming a career is associated with physical, emotional, and financial hardship, with caregivers often experiencing a maelstrom of emotions as they struggle to understand what has happened to their loved one. While the burden of caring for young people with first-episode psychosis (FEP) has been well documented, much less is known about how carers develop the strength and resilience to continue caring. This qualitative study aimed to understand the experience of 20 first time primary caregivers of young adults with FEP. Most caregivers were female (85%, n = 17) and parents (85%, n = 17). The average length of involvement as a caregiver at an FEP service was 14.5 months. Six main themes were identified in the data, highlighting the carers' experience in supporting young adults with FEP. Caregiving is a burdensome responsibility and is characterized as a roller coaster and unpredictable experience. Caregivers often feel responsible for the young person's illness; however, eventually most come to terms with the changes that have occurred in the young person with FEP. As a consequence of the illness, the relationship between caregiver and care recipient frequently becomes closer and deeper, although it is important that they both maintain hope for the future. These findings provide important insights into the experiences of first-time caregivers of young people with FEP, with direct implications for improving the information and support given to caregivers by FEP services, as well as the development of interventions that effectively address the unique challenges caregivers face following the onset of FEP. PMID- 19679718 TI - Cycliophoran dwarf males break the rule: high complexity with low cell numbers. AB - Complexity of metazoan bodyplans is commonly assumed to be correlated to the absolute number of cells and the number of cell types present in a species (1). Sexually mature individuals of the smallest free-living animals have a minimum of several hundred somatic cells, and only secondarily simplified parasitic or commensal species range below this threshold. Males of the two hitherto described representatives of the phylum Cycliophora (2), with a body length of about 40 microm, are among the smallest existing free-living metazoans, yet they exhibit an amazingly complex bodyplan. Herein, we show that only a few dozen cells account for this complexity. We conclude therefore that metazoan complexity is not obligatorily correlated with body size or with the overall cell number of an individual. Accordingly, the earliest multicellular animals on Earth, which most probably were small individuals, may have had more complex bodyplans than commonly assumed. PMID- 19679719 TI - Nutrient uptake by marine invertebrates: cloning and functional analysis of amino acid transporter genes in developing sea urchins (Strongylocentrotus purpuratus). AB - Transport of amino acids from low concentrations in seawater by marine invertebrates has been extensively studied, but few of the genes involved in this physiological process have been identified. We have characterized three amino acid transporter genes cloned from embryos of the sea urchin Strongylocentrotus purpuratus. These genes show phylogenetic proximity to classical amino acid transport systems, including Gly and B0+, and the inebriated gene (INE). Heterologous expression of these genes in frog oocytes induced a 40-fold increase in alanine transport above endogenous levels, demonstrating that these genes mediate alanine transport. Antibodies specific to one of these genes (Sp-AT1) inhibited alanine transport, confirming the physiological activity of this gene in larvae. Whole-mount antibody staining of larvae revealed expression of Sp-AT1 in the ectodermal tissues associated with amino acid transport, as independently demonstrated by autoradiographic localization of radioactive alanine. Maximum rates of alanine transport increased 6-fold during early development, from embryonic to larval stages. Analysis of gene expression during this developmental period revealed that Sp-AT1 transcript abundance remained nearly constant, while that of another transporter gene (Sp-AT2) increased 11-fold. The functional characterization of these genes establishes a molecular biological basis for amino acid transport by developmental stages of marine invertebrates. PMID- 19679720 TI - Formation of the apical flaps in nematocysts of sea anemones (cnidaria: actiniaria). AB - Using scanning and transmission electron microscopy, we studied formation of the structure at the apical end of sea anemone nematocysts through which the tubule everts at discharge. In anemones of the genus Metridium, we found that each of the three solid triangular apical flaps comprises two layers that are continuous with those of the capsule wall: the electron-lucent inner layer is bound to the electron-dense outer layer. The two-layer structure is obvious in some discharged capsules in which, perhaps due to fixation, the layers part at the flap's periphery. Before the nematocyst discharges, a channel leads from a pore at the tip of the joined flaps into the lumen of the inverted tubule. The thin laminate layer that coats each flap lines the channel. The base of the nematocyst tubule adheres to the capsule wall near the capsule's apical end, and a branch of the tubule underlies part of the laminate layer that coats the flaps. Thus the tubule is not continuous with the capsule wall but structurally separate from it. This helps reconcile differences in understanding of the number of layers constituting the capsule wall, and makes clear that the tubule should be considered part of the capsule contents. PMID- 19679721 TI - Ultrastructure of the retinal synapses in cubozoans. AB - Cubomedusae (box jellyfish) are well known for strong directional swimming, rapid responses to visual stimuli, and complex lensed eyes comparable to those of more advanced multicellular animals. They possess a total of 24 eyes that are of four morphologically different types, yet little is known about the neural organization of their eyes. The eyes are located on ganglion-like structures called rhopalia. Each of the four rhopalia contains an upper and a lower lensed eye (with a cornea, lens, and retina), two pit ocelli, and two slit ocelli. Transmission electron microscopy was used to examine the synaptic morphology of the eyes and pacemaker region of four species of cubozoans (Tamoya haplonema, Carybdea marsupialis, Tripedalia cystophora, and Chiropsalmus quadrumanus). Invaginated synapses were found in all four species, but only in the upper and lower lensed eyes. Density measurements indicated that the invaginated synapses were located close to the basal region of photoreceptor cells, and size differences of invaginated synapses were observed between the upper and lower lensed eyes, as well as between species. Four additional types of chemical synapses-clear unidirectional, dense-core unidirectional, clear bidirectional, and clear and dense-core bidirectional-were also observed in the rhopalia. The invaginated synapses of the lensed eyes may be useful as markers to help sort out the neural circuitry in the retinal region of these complex cubomedusan eyes. PMID- 19679722 TI - Serotonergic modulation of crayfish hindgut. AB - The crayfish hindgut is a morphologically differentiated tube that varies along its length in the distribution of muscles and glands, contractile properties, serotonergic innervation, patterns of 5-HT receptor expression, and sensitivity to serotonin (5-HT). Anatomical differences divide the hindgut into five distinct segments along its length. Spontaneous pulsatile contractions produced by the isolated hindgut decrease in force and increase in frequency along the anterior posterior axis. Central input to the hindgut comes from a large cluster of 5-HT immunoreactive neurons in the terminal abdominal ganglion that form a large nerve plexus on the hindgut. 5-HT(1alpha) and 5-HT(2beta) receptors vary in their distribution along the hindgut, and are associated with longitudinal and circular muscles and with axon collaterals of the 5-HT-immunoreactive neurons. Application of 30 nmol l(-1) to 1 mumol l(-1) 5-HT to rostral, middle, or caudal sections of hindgut produced tension changes that varied with the concentration and section. 5-HT also initiated antiperistaltic waves in the posterior hindgut. These results indicate that 5-HT is an important neuromodulator for initiating contractions and coordinating activity in the different functional compartments along the rostral to-caudal axis of the hindgut. PMID- 19679723 TI - Species-specific vulnerability of benthic marine embryos of congeneric snails (Haminoea spp.) to ultraviolet radiation and other intertidal stressors. AB - We used field surveys and multi-factorial experiments to examine synergistic effects of ultraviolet radiation (UVR) and low tide conditions on the embryonic mortality of two bubble-shell snail species that deposit gelatinous egg masses in intertidal mudflats: Haminoea zelandiae from New Zealand, and Haminoea vesicula from Washington, USA. Egg masses of both species were predominantly found in shallow pools at low tide, and a substantial proportion of both were found in sunny as well as shaded microhabitats. Both exposure to sun and desiccation led to increased embryonic mortality for naturally deposited egg masses of H. zelandiae compared to those that were shaded or submerged. For H. vesicula, although mortality was double for embryos within desiccated egg masses, there was no additional mortality due to sun exposure. In manipulative experiments, UVR and low tide conditions increased embryonic mortality for both species; however, H. zelandiae appeared to be more vulnerable to UVR, whereas H. vesicula was particularly vulnerable to desiccation. Simulated tidal pool conditions significantly increased mortality only for H. vesicula. These results suggest an important role of species-specific differences in vulnerability to different stressors, even for ecologically similar congeners; here, these differences may be related to development time or egg mass characteristics. PMID- 19679724 TI - Linking thermal tolerances and biogeography: Mytilus edulis (L.) at its southern limit on the east coast of the United States. AB - Temperature is a major factor contributing to the latitudinal distribution of species. In the Northern Hemisphere, a species is likely to be living very close to its upper thermal tolerance limits at the southern limit of its biogeographic range. With global warming, this southern limit is expected to shift poleward. Moreover, intertidal ecosystems are expected to be especially strongly affected, mostly due to their large daily and seasonal variations in temperature and exposure. Hence, these are model systems in which to conduct experiments examining the ecological effects of climate change. In this study we determined the upper lethal thermal limits, for both air and water, of the blue mussel Mytilus edulis via laboratory experiments. Tolerances vary seasonally, with a difference between media of 0.7 degrees C in June and 4.8 degrees C in November, as well as a decrease with multiple exposures. Measured lethal limits were then compared to field measurements of environmental temperature and concurrent measurements of mortality rates. Field results indicate that mortality in the intertidal occurs at rates expected from laboratory responses to elevated temperature. Hindcasts, retrospective analyses of historical data, indicate that high rates of mortality have shifted 51 and 42 days earlier in Beaufort, North Carolina, and Oregon Inlet, North Carolina, respectively, between 1956 and 2007. The combined data suggest that the historical southern limit of M. edulis near Cape Hatteras, North Carolina, is indeed the result of intolerance to high temperature, and that this range edge is shifting poleward in a manner indicative of global warming. PMID- 19679725 TI - Estuarine-scale genetic variation in the polychaete hobsonia Florida (ampharetidae; annelida) in long island sound and relationships to pleistocene glaciations. AB - Modern patterns of genetic variation in northern estuarine species are likely shaped by both historical and contemporary processes. Many studies have demonstrated the predominant roles of Pleistocene glaciations, life-history traits, or a combination of both in shaping patterns of population differentiation across a broad range of animal species, yet comparatively few have investigated such questions in benthic invertebrates. We examined fine-scale patterns of genetic differentiation among populations in the ampharetid polychaete Hobsonia florida in Long Island Sound (LIS) relative to its life history traits and the geologic history of the region. Mitochondrial DNA sequencing of cytochrome oxidase I (COI) revealed 15 unique haplotypes among four populations in LIS and two outgroups. Populations in LIS exhibited the greatest haplotypic diversity compared to outgroup populations. Analysis of molecular variance revealed a highly significant level of genetic heterogeneity (P < 0.001) in LIS, with almost 40% of the variance attributed to differences among populations. Population differentiation was further revealed by highly significant fixation indices for all pairwise comparisons. Analysis using a mtDNA COI molecular clock estimated that haplotype divergence ranged from about 73,000 to 366,000 years before present. Our results suggest that the observed population genetic structure of H. florida in LIS has been strongly influenced by a series of glacially mediated vicariance events, leading to periodic population isolation and mixing, further mediated by low dispersal and specialized habitat requirements. PMID- 19679726 TI - Idiopathic lesions and visual deficits in the american lobster (Homarus americanus) from Long Island Sound, NY. AB - In 1999, a mass mortality of the American lobster (Homarus americanus) occurred in western Long Island Sound (WLIS). Although the etiology of this event remains unknown, bottom water temperature, hypoxia, heavy metal poisoning, and pesticides are potential causal factors. Lobsters from WLIS continue to display signs of morbidity, including lethargy and cloudy grey eyes that contain idiopathic lesions. As the effect of these lesions on lobster vision is unknown, we used electroretinography (ERG) to document changes in visual function in lobsters from WLIS, while using histology to quantify the extent of physical damage. Seventy three percent of lobsters from WLIS showed damage to photoreceptors and optic nerve fibers, including necrosis, cellular breakdown, and hemocyte infiltration in the optic nerves, rhabdoms, and ommatidia. Animals with more than 15% of their photoreceptors exhibiting damage also displayed markedly reduced responses to 10 ms flashes of a broad-spectrum white light. Specifically, maximum voltage (Vmax) responses were significantly lower and occurred at a lower light intensity compared to responses from lobsters lacking idiopathic lesions. Nearly a decade after the 1999 mortality event, lobsters from WLIS still appear to be subjected to a stressor of unknown etiology that causes significant functional damage to the eyes. PMID- 19679727 TI - Your hospital was a 'clan', now it's 'rational'. PMID- 19679728 TI - Beyond the trauma registry. PMID- 19679729 TI - Quality improvement for doctors - it's essential. PMID- 19679730 TI - Dematuration. PMID- 19679731 TI - Age-related macular degeneration and its effect on quality of life. PMID- 19679732 TI - Hypoparathyroidism and coeliac disease: a potentially dangerous combination. PMID- 19679733 TI - Richard III: a study in medical misrepresentation. PMID- 19679734 TI - An alternative approach to medical genetics based on modern evolutionary biology. Part 2: retroviral symbiosis. PMID- 19679735 TI - From cultural cohesion to rules and competition: the trajectory of senior management culture in English NHS hospitals, 2001-2008. AB - OBJECTIVES: To assess changes in senior management culture in English NHS hospitals between 2001 and 2008. DESIGN: Longitudinal study comprising cross sectional surveys of board levels managers at three time-points using a validated culture measurement instrument - the Competing Values Framework. SETTING: English NHS hospital trusts. MAIN OUTCOMES: There has been a decline in 'Clan' and an increase in more competitive 'Rational' cultures among senior management teams in NHS hospital trusts during the period 2001-2008. CONCLUSIONS: Government policy for the NHS has espoused the desirability of competition, particularly since 2002. Our data suggest that corresponding changes in the cultures of senior management teams in NHS hospital trusts is beginning to occur, with the expectation that these organizations will pursue more competitive strategies. PMID- 19679736 TI - The gender imbalance in academic medicine: a study of female authorship in the United Kingdom. AB - OBJECTIVES: A shortfall exists of female doctors in senior academic posts in the United Kingdom. Career progression depends on measures of esteem, including publication in prestigious journals. This study investigates gender differences in first and senior authorship in six peer-reviewed British journals and factors that are associated with publication rates. DESIGN AND MAIN OUTCOME MEASURES: Data was collected on United Kingdom first and senior authors who had published in the British Medical Journal, Lancet, British Journal of Surgery, Gut, British Journal of Obstetrics and Gynaecology and the Archives of Diseases in Childhood. Authorship and gender were quantified for 1970, 1980, 1990, 2000 and 2004 (n=6457). In addition, selected questions from the Athena Survey of Science Engineering and Technology (ASSET2006), web-based doctor's self-report of publications were also analysed (n=1162). RESULTS: Female first authors increased from 10.5% in 1970 to 36.5% in 2004 (p<0.001) while female senior authors only increased from 12.3% to 16.5% (p=0.046). Within individual journals, the largest rise was in British Journal of Obstetric and Gynaecology with 4.5- and 3-fold increases for first and senior authors, respectively. In contrast, female senior authors marginally declined in Gut and Lancet by 2.8% and 2.2%, respectively. ASSET2006 identified that female respondents who were parents were less likely to have publications as sole (p=0.02) and joint authors (p<0.001) compared to male respondents. Female respondents with care responsibilities for parents/partner also had less publications as lead authors compared to those without carer responsibilities (p<0.001). CONCLUSION: The increase in UK female first authors is encouraging. In contrast, there is considerable lag and in some specialties a decline in female senior authors. Factors that could narrow the gender gap in authorship should be sought and addressed. PMID- 19679737 TI - The introduction of 'chemotherapy' using arsphenamine - the first magic bullet. PMID- 19679740 TI - Impact factor and its role in academic promotion: a statement adopted by the International Respiratory Journal Editors Roundtable. PMID- 19679739 TI - Frailty and the degradation of complex balance dynamics during a dual-task protocol. AB - BACKGROUND: Balance during quiet stance involves the complex interactions of multiple postural control systems, which may degrade with frailty. The complexity of center of pressure (COP) dynamics, as quantified using multiscale entropy (MSE), during quiet standing is lower in older adults, especially those with falls. We hypothesized that COP dynamics from frail elderly individuals demonstrate less complexity than those from nonfrail elderly controls; complexity decreases when performing a dual task; and postural complexity during quiet standing is independent of other conventional correlates of balance control, such as age and vision. METHODS: We analyzed data from a population-based study of community-dwelling older adults. Frailty phenotype (nonfrail, prefrail, or frail) was determined for 550 participants (age 77.9 +/- 5.5 years). COP excursions were quantified for 10 trials of 30 seconds each. Participants concurrently performed a serial subtraction task in half of the trials. Complexity of balance dynamics was quantified using MSE. Root-mean-square sway amplitude was also computed. RESULTS: Of the 550, 38% were prefrail and 9% were frail. Complexity of the COP dynamics in the anteroposterior direction was lower in prefrail (8.78 +/- 1.91 [mean +/- SD]) and frail (8.38 +/- 2.13) versus nonfrail (9.20 +/- 1.74) groups (p < .001). Complexity reduced by a comparable amount in all three groups while performing the subtraction task (p < .001). Quiet standing complexity was independently associated with frailty after adjusting for covariates related to balance while sway amplitude was not. CONCLUSION: Cognitive distractions during standing may further compromise balance control in frail individuals, leading to an increased risk of falls. PMID- 19679742 TI - Aging affects the cardiovascular responses to cold stress in humans. AB - Cardiovascular-related mortality peaks during cold winter months, particularly in older adults. Acute physiological responses, such as increases in blood pressure, in response to cold exposure may contribute to these associations. To determine whether the blood pressure-raising effect (pressor response) of non-internal body temperature-reducing cold stress is greater with age, we measured physiological responses to 20 min of superficial skin cooling, via water-perfused suit, in 12 younger [25 +/- 1 (SE) yr old] and 12 older (65 +/- 2 yr old) adults. We found that superficial skin cooling elicited an increase in blood pressure from resting levels (pressor response; P < 0.05) in younger and older adults. However, the magnitude of this pressor response (systolic and mean blood pressure) was more than twofold higher in older adults (P < 0.05 vs. younger adults). The magnitude of the pressor response was similar at peripheral (brachial) and central (estimated in the aorta) measurement sites. Regression analysis revealed that aortic pulse wave velocity, a measure of central arterial stiffness obtained before cooling, was the best predictor of the increased pressor response to superficial skin cooling in older adults, explaining approximately 63% of its variability. These results indicate that there is a greater pressor response to non-internal body temperature-reducing cold stress with age in humans that may be mediated by increased levels of central arterial stiffness. PMID- 19679743 TI - The phenotype and potential origin of nestin+ cardiac myocyte-like cells following infarction. AB - Nestin+ cardiac myocyte-like cells were detected in the peri-infarct/infarct region of the ischemically damaged heart. The present study was undertaken to elucidate the phenotype and potential origin of nestin+ cardiac myocyte-like cells and identify stimuli implicated in their appearance. In the infarcted human and rat heart, nestin+ cardiac myocyte-like cells were morphologically and structurally immature, exhibited a desmin-immunoreactive striated phenotype, expressed the beta(1)-adrenergic receptor, and associated with an aberrant pattern of connexin-43 expression and/or organization. Nestin+ cardiac myocyte like cells were detected 24 h postischemic injury and persisted in the infarcted rat heart for 9 mo. In the normal rat heart, cardiac progenitor transcriptional factors Nkx2.5/GATA4 were detected in a subpopulation of nestin+ neural stem cells. Following an ischemic insult, nestin+/Nkx2.5+ neural stem cells migrated to the peri-infarct/infarct region and appeared to be in a primordial state of differentiation to a nestin+ cardiac myocyte-like cell. The exposure of adult male rats to normobaric hypoxia (12% O2) for 10 days failed to promote the appearance of nestin+ cardiac myocyte-like cells. Following osmotic pump delivery of isoproterenol to normal adult rats, nestin+ cardiac myocyte-like cells were detected, albeit the response was modest and secondary to tissue loss. Thus ischemia-induced appearance of nestin+ cardiac myocyte-like cells apparently represents an adaptive response to heal the infarcted heart. Nkx2.5/GATA4 expression in a subpopulation of resident neural stem cells provides the appropriate phenotype for their potential differentiation to a nestin+ cardiac myocyte-like cell. PMID- 19679741 TI - Pressed for time: the circadian clock and hypertension. AB - Hypertension is a major risk factor for cardiovascular disease and death. The "silent" rise of blood pressure that occurs over time is largely asymptomatic. However, its impact is deafening-causing and exacerbating cardiovascular disease, end-organ damage, and death. The present article addresses recent observations from human and animal studies that provide new insights into how the circadian clock regulates blood pressure, contributes to hypertension, and ultimately evolves vascular disease. Further, the molecular components of the circadian clock and their relationship with locomotor activity, metabolic control, fluid balance, and vascular resistance are discussed with an emphasis on how these novel, circadian clock-controlled mechanisms contribute to hypertension. PMID- 19679744 TI - Influence of central command and muscle afferent activation on anterior cerebral artery blood velocity responses to calf exercise in humans. AB - The purpose of the present study was to determine the relative importance of peripheral feedback from mechanically (mechanoreflex) and metabolically (metaboreflex) sensitive muscle afferents and central signals arising from higher centers (central command) to the exercise-induced increases in regional cerebral perfusion. To accomplish this, anterior cerebral artery (ACA) mean blood velocity (V(mean)) responses were assessed during sustained and rhythmic passive calf muscle stretch (mechanoreflex), volitional calf exercise (mechanoreflex, metaboreflex, and central command), and electrically stimulated calf exercise (mechanoreflex and metaboreflex but no central command) at 35% of maximum voluntary contraction (n = 16). In addition, a period of postexercise muscle ischemia (PEMI) was used to isolate the metaboreflex. Blood pressure, cardiac output, and the end-tidal partial pressure of carbon dioxide (Pet(CO2)) were also measured. ACA V(mean) was unchanged from rest during either sustained or rhythmic calf muscle stretch (P > 0.05). However, ACA V(mean) was increased from rest during both isometric (+15 +/- 1%) and rhythmic (+15 +/- 2%, voluntary exercise P < 0.05) but remained unchanged during stimulated exercise (P > 0.05). Isometric and rhythmic exercise-induced increases in blood pressure and cardiac output were similar during voluntary and stimulated exercise (P > 0.05 between conditions). Blood pressure remained elevated during PEMI after all exercise conditions (P < 0.05 vs. rest), whereas cardiac output and ACA V(mean) were not different from rest (P > 0.05). Pet(CO2) was unchanged from rest throughout. These data suggest that selective activation of skeletal muscle afferents (i.e., stretch, PEMI, or stimulated exercise) does not increase ACA V(mean) and that increases in ACA V(mean) during volitional contractions of an exercising calf muscle are dependent on the presence of central command. PMID- 19679745 TI - Moderate intensity of regular exercise improves cardiac SR Ca2+ uptake activity in ovariectomized rats. AB - The impact of regular exercise in protecting cardiac deteriorating results of female sex hormone deprivation was evaluated by measuring changes in intracellular Ca2+ removal activity of sarcoplasmic reticulum (SR) in ovariectomized rats following 9-wk treadmill running exercise at moderate intensity. Despite induction of cardiac hypertrophy in exercised groups of both sham-operated and ovariectomized rats, exercise training had no effect on SR Ca2+ uptake and SR Ca(2+)-ATPase (SERCA) in hormone intact rat heart. However, exercise training normalized the suppressed maximum SR Ca2+ uptake and SERCA activity in ovariectomized rat heart. While exercise training normalized the leftward shift in pCa (-log[Ca2+])-SR Ca2+ uptake relation in ovariectomized rats, no effect was detected in exercised sham-operated rats. Similar phenomena were also observed on SERCA and on phospholamban (PLB) phosphorylation levels; exercise training in ovariectomized rats enhanced SERCA expression to reach the level as that in sham-operated rats, in which there were no differences in SERCA and phospho-PLB levels between sedentary and exercised groups. In addition, the reduction in phospho-Thr(17) PLB in myocardium of ovariectomized rats was abolished by exercise training. These results showed that regular exercise maintains the molecular activation of cardiac SR Ca2+ uptake under normal physiological conditions and is able to induce a protective impact on cardiac SR Ca2+ uptake in ovarian sex hormone-deprived status. PMID- 19679746 TI - Galvanic vestibular stimulation counteracts hypergravity-induced plastic alteration of vestibulo-cardiovascular reflex in rats. AB - Recent data from our laboratory demonstrated that, when rats are raised in a hypergravity environment, the sensitivity of the vestibulo-cardiovascular reflex decreases. In a hypergravity environment, static input to the vestibular system is increased; however, because of decreased daily activity, phasic input to the vestibular system may decrease. This decrease may induce use-dependent plasticity of the vestibulo-cardiovascular reflex. Accordingly, we hypothesized that galvanic vestibular stimulation (GVS) may compensate the decrease in phasic input to the vestibular system, thereby preserving the vestibulo-cardiovascular reflex. To examine this hypothesis, we measured horizontal and vertical movements of rats under 1-G or 3-G environments as an index of the phasic input to the vestibular system. We then raised rats in a 3-G environment with or without GVS for 6 days and measured the pressor response to linear acceleration to examine the sensitivity of the vestibulo-cardiovascular reflex. The horizontal and vertical movement of 3-G rats was significantly less than that of 1-G rats. The pressor response to forward acceleration was also significantly lower in 3-G rats (23 +/- 1 mmHg in 1-G rats vs. 12 +/- 1 mmHg in 3-G rats). The pressor response was preserved in 3-G rats with GVS (20 +/- 1 mmHg). GVS stimulated Fos expression in the medial vestibular nucleus. These results suggest that GVS stimulated vestibular primary neurons and prevent hypergravity-induced decrease in sensitivity of the vestibulo-cardiovascular reflex. PMID- 19679747 TI - Dietary quercetin supplementation is not ergogenic in untrained men. AB - Quercetin supplementation increases muscle oxidative capacity and endurance in mice, but its ergogenic effect in humans has not been established. Our study investigates the effects of short-duration chronic quercetin supplementation on muscle oxidative capacity; metabolic, perceptual, and neuromuscular determinants of performance in prolonged exercise; and cycling performance in untrained men. Using a double-blind, pretest-posttest control group design, 30 recreationally active, but not endurance-trained, young men were randomly assigned to quercetin and placebo groups. A noninvasive measure of muscle oxidative capacity (phosphocreatine recovery rate using magnetic resonance spectroscopy), peak oxygen uptake (Vo(2peak)), metabolic and perceptual responses to submaximal exercise, work performed on a 10-min maximal-effort cycling test following the submaximal cycling, and voluntary and electrically evoked strength loss following cycling were measured before and after 7-16 days of supplementation with 1 g/day of quercetin in a sports hydration beverage or a placebo beverage. Pretreatment to-posttreatment changes in phosphocreatine recovery time constant, Vo(2peak,) substrate utilization, and perception of effort during submaximal exercise, total work done during the 10-min maximal effort cycling trial, and voluntary and electrically evoked strength loss were not significantly different (P > 0.05) in the quercetin and placebo groups. Short duration, chronic dietary quercetin supplementation in untrained men does not improve muscle oxidative capacity; metabolic, neuromuscular and perceptual determinants of performance in prolonged exercise; or cycling performance. The null findings indicate that metabolic and physical performance consequences of quercetin supplementation observed in mice should not be generalized to humans. PMID- 19679748 TI - Lung function in developing lambs: is it affected by preterm birth? AB - Children born before term often have reduced lung function, but the effects of preterm birth alone are difficult to determine owing to iatrogenic factors such as mechanical ventilation. Our objective was to determine the effects of preterm birth alone on airway resistance, airway reactivity, and ventilatory heterogeneity as an index of intrapulmonary gas mixing. Preterm birth was induced in sheep 12 days before term; controls were born at term ( approximately 147 days). Lung function was assessed at 8 wk postterm. To assess medium-large airway function we measured airway resistance and reactivity to carbachol. Multiple breath N(2) washout (MBW) was used to assess ventilatory heterogeneity in conducting (S(cond)) and acinar (S(acin)) airways. Baseline airway resistance and responsiveness to carbachol were similar in preterm and term lambs. Airway responsiveness to carbachol was greater in females than males (P < 0.05), and baseline airway resistance tended to be higher in females than males (P = 0.06). There were no significant differences in ventilatory heterogeneity between preterm and term lambs; for all animals combined, mean S(acin) was 0.29 +/- 0.05 liter(-1) and S(cond) was 0.26 +/- 0.03 liter(-1). Males had significantly higher S(cond) than females, indicating poorer gas mixing in small conducting airways; there was no sex difference in S(acin). We conclude that preterm birth per se in lambs does not affect baseline airway resistance, airway responsiveness, or ventilatory heterogeneity as measured by MBW. The observed sex-related differences in airway responsiveness and ventilatory heterogeneity in the conducting airways could help explain sex differences in lung function observed in humans. PMID- 19679749 TI - Placental malarial infection as a risk factor for hypertensive disorders during pregnancy in Africa: a case-control study in an urban area of Senegal, West Africa. AB - In tropical countries, malaria and hypertension are common diseases of pregnancy. They have physiopathologic similarities such as placental ischemia, endothelial dysfunction, and production of proinflammatory cytokines. Recent findings suggested their possible link. The authors conducted a case-control study to explore the relation between malaria and hypertension at Guediawaye, a hypoendemic malarial setting in Senegal. Cases were pregnant women admitted to the delivery unit for hypertension. Controls were pregnant women admitted for normal delivery, without any history of hypertension or proteinuria during the present pregnancy. Malarial infection was determined by placental tissue examination. From January to December 2002, 77 cases of gestational hypertension, 113 cases of preeclampsia, 59 cases of eclampsia, and 241 controls were enrolled. Placental malarial infection (PMI) was present in 14 cases (6.3%) and in 15 controls (6.2%). The prevalence of PMI was 4.6% for eclampsia, 4.0% for preeclampsia, and 11.6% for gestational hypertension. In multivariate analysis, PMI appeared to be an independent risk factor for gestational hypertension (adjusted odds ratio = 2.7, 95% confidence interval: 1.0, 7.6). The authors found an association between PMI and nonproteinuric hypertension in women living in a malaria-hypoendemic area. The exact significance of such relation should be clarified in further studies in different settings of malarial endemicity. PMID- 19679751 TI - The need for validation of statistical methods for estimating respiratory virus attributable hospitalization. AB - Public policy regarding influenza has been based largely on the burden of hospitalization estimated through ecologic studies applying increasingly sophisticated statistical methods to administrative databases. None are known to have been validated by observational studies. The authors illustrated how 6 commonly applied statistical methods estimate virus-attributable hospitalization of children 6-23 months of age and compared the estimates with results obtained from a prospective study using virologic assessment. The proportions of pneumonia and influenza and of bronchiolitis hospitalizations attributable to respiratory syncytial virus and/or influenza were derived by using Serfling regression, periseason differences, Poisson regression with log link, negative binomial regression with identity link, and a Box-Jenkins transfer function. No method provided accurate or consistent estimates for both viruses and outcomes. Virus attributable hospitalization estimates varied widely between statistical methods and between seasons, with greater between-season variation for admissions attributed to influenza compared with respiratory syncytial virus. Sophistication of statistical methods may have been interpreted as assurance that results are more accurate. Without validation against epidemiologic data, with viral etiology confirmed in individual patients, the accuracy of statistical methods in ecologic studies is simply not known. Until these methods are validated, their methodological limitations should be made explicit and proxy estimates used cautiously in guiding public policy. PMID- 19679750 TI - Strategies for pandemic and seasonal influenza vaccination of schoolchildren in the United States. AB - Vaccinating school-aged children against influenza can reduce age-specific and population-level illness attack rates. Using a stochastic simulation model of influenza transmission, the authors assessed strategies for vaccinating children in the United States, varying the vaccine type, coverage level, and reproductive number R (average number of secondary cases produced by a typical primary case). Results indicated that vaccinating children can substantially reduce population level illness attack rates over a wide range of scenarios. The greatest absolute reduction in influenza illness cases per season occurred at R values ranging from 1.2 to 1.6 for a given vaccine coverage level. The indirect, total, and overall effects of vaccinating children were strong when transmission intensity was low to intermediate. The indirect effects declined rapidly as transmission intensity increased. In a mild influenza season (R = 1.1), approximately 19 million influenza cases could be prevented by vaccinating 70% of children. At most, nearly 100 million cases of influenza illness could be prevented, depending on the proportion of children vaccinated and the transmission intensity. Given the current worldwide threat of novel influenza A (H1N1), with an estimated R of 1.4 1.6, health officials should consider strategies for vaccinating children against novel influenza A (H1N1) as well as seasonal influenza. PMID- 19679752 TI - Pistillata--duplications as a mode for floral diversification in (Basal) asterids. AB - Basal asterid families, and to a lesser extent the asterids as a whole, are characterized by a high variation in petal and stamen morphology. Moreover, the stamen number, the adnation of stamens to petals, and the degree of sympetaly vary considerably among basal asterid taxa. The B group genes, members of the APETALA3 (AP3) and PISTILLATA (PI) gene lineages, have been shown to specify petal and stamen identities in several core eudicot species. Duplicate genes in these lineages have been shown in some cases to have diversified in their function; for instance in Petunia, a PI paralog is required for the fusion of stamens to the corolla tube, illustrating that such genes belonging to this lineage are not just involved in specifying the identity of the stamens and petals but can also specify novel floral morphologies. This motivated us to study the duplication history of class B genes throughout asterid lineages, which comprise approximately one-third of all flowering plants. The evolutionary history of the PI gene subfamily indicates that the two genes in Petunia result from an ancient duplication event, coinciding with the origin of core asterids. A second duplication event occurred before the speciation of basal asterid Ericales families. These and other duplications in the PI lineage are not correlated with duplications in the AP3 lineage. To understand the molecular evolution of the Ericales PI genes after duplication, we have described their expression patterns using reverse transcription polymerase chain reaction and in situ hybridization, reconstructed how selection shaped their protein sequences and tested their protein interaction specificity with other class B proteins. We find that after duplication, PI paralogs have acquired multiple different expression patterns and negative selective pressure on their codons is relaxed, whereas substitutions in sites putatively involved in protein-protein interactions show positive selection, allowing for a change in the interaction behavior of the PI paralogs after duplication. Together, these observations suggest that the asterids have preferentially recruited PI duplicate genes to diverse and potentially novel roles in asterid flower development. PMID- 19679753 TI - Age-related accumulation of mutations supports a replication-dependent mechanism of spontaneous mutation at tandem repeat DNA Loci in mice. AB - Expanded simple tandem repeat (ESTR) loci belong to the class of highly unstable loci in the mouse genome. The mechanisms underlying the very high spontaneous instability at these loci still remain poorly understood. Using single-molecule polymerase chain reaction, here we have compared the pattern of mutation accumulation in tissues with different proliferation capacities in male mice of age 12, 26, 48, and 96 weeks. In the nonproliferating brain, we did not observe any measurable age-related accumulation of ESTR mutations. In contrast, a highly elevated frequency of ESTR mutation was detected in the sperm samples taken from old mice; similar changes were also observed in the bone marrow tissue. The spectra of ESTR mutations accumulated in all tissues of young and old mice did not significantly differ. Taken together, these data clearly imply that spontaneous ESTR mutations occur almost exclusively in replication-proficient cells. To gain further insights into the mechanisms of ESTR mutation, we developed a stochastic model of age-related mutation accumulation. The observed spectra of ESTR mutants accumulated in the brain and sperm were fairly accurately approximated assuming the values of ESTR mutation rate, ranging from 0.01 to 0.04 per cell division. As these estimates dramatically exceed those for protein coding genes and microsatellite loci, our data therefore suggest that ESTRs represent one of the most unstable loci in the mammalian genome. The results of our study also imply that ESTR loci can be regarded as a class of expanded microsatellites, with the mechanism of spontaneous mutation most probably attributed to replication slippage. PMID- 19679754 TI - Molecular evolution of GYPC: evidence for recent structural innovation and positive selection in humans. AB - GYPC encodes two erythrocyte surface sialoglycoproteins in humans, glycophorin C and glycophorin D (GPC and GPD), via initiation of translation at two start codons on a single transcript. The malaria-causing parasite Plasmodium falciparum uses GPC as a means of invasion into the human red blood cell. Here, we examine the molecular evolution of GYPC among the Hominoidea (Greater and Lesser Apes) and also the pattern of polymorphism at the locus in a global human sample. We find an excess of nonsynonymous divergence among species that appears to be caused solely by accelerated evolution of GYPC in the human lineage. Moreover, we find that the ability of GYPC to encode both GPC and GPD is a uniquely human trait, caused by the evolution of the GPC start codon in the human lineage. The pattern of polymorphism among humans is consistent with a hitchhiking event at the locus, suggesting that positive natural selection affected GYPC in the relatively recent past. Because GPC is exploited by P. falciparum for invasion of the red blood cell, we hypothesize that selection for evasion of P. falciparum has caused accelerated evolution of GYPC in humans (relative to other primates) and that this positive selection has continued to act in the recent evolution of our species. These data suggest that malaria has played a powerful role in shaping molecules on the surface of the human red blood cell. In addition, our examination of GYPC reveals a novel mechanism of protein evolution: co-option of untranslated region (UTR) sequence following the formation of a new start codon. In the case of human GYPC, the ancestral protein (GPD) continues to be produced through leaky translation. Because leaky translation is a widespread phenomenon among genes and organisms, we suggest that co-option of UTR sequence may be an important source of protein innovation. PMID- 19679758 TI - The context counts: congruent learning and testing environments prevent memory retrieval impairment following stress. AB - Stress before retention testing impairs memory, whereas memory performance is enhanced when the learning context is reinstated at retrieval. In the present study, we examined whether the negative impact of stress before memory retrieval can be attenuated when memory is tested in the same environmental context as that in which learning took place. Subjects learned a 2-D object location task in a room scented with vanilla. Twenty-four hours later, they were exposed to stress or a control condition before memory for the object location task was assessed in a cued-recall test, either in the learning context or in a different context (unfamiliar room without the odor). Stress impaired memory when assessed in the unfamiliar context, but not when assessed in the learning context. These results suggest that the detrimental effects of stress on memory retrieval can be abolished when a distinct learning context is reinstated at test. PMID- 19679759 TI - Oxytocin enhances the perception of biological motion in humans. AB - Evidence suggests that intranasally administered oxytocin modulates several social cognitive and emotional processes in humans. In this study, we investigated the effect of oxytocin on the perception of biological motion (a walking character) and nonbiological motion (a rotating shape). The participants were 20 healthy volunteers who observed moving dots embedded among a cloud of noise (mask) dots. Sensitivity (d') for motion detection was determined after the administration of oxytocin and placebo. The results showed that oxytocin (relative to placebo) administration increased sensitivity to biological motion but not to nonbiological motion. These results suggest that oxytocin specifically modulates the perception of socially relevant stimuli. PMID- 19679760 TI - Activation of right parietal cortex during memory retrieval of nonlinguistic auditory stimuli. AB - In neuroimaging studies, the left ventral posterior parietal cortex (PPC) is particularly active during memory retrieval. However, most studies have used verbal or verbalizable stimuli. We investigated neural activations associated with the retrieval of short, agrammatical music stimuli (Blackwood, 2004), which have been largely associated with right hemisphere processing. At study, participants listened to music stimuli and rated them on pleasantness. At test, participants made old/new recognition judgments with high/low confidence ratings. Right, but not left, ventral PPC activity was observed during the retrieval of these music stimuli. Thus, rather than indicating a special status of left PPC in retrieval, both right and left ventral PPC participate in memory retrieval, depending on the type of information that is to be remembered. PMID- 19679761 TI - Behavioral and neural correlates of memory selection and interference resolution during a digit working memory task. AB - Neuroimaging studies have shown the involvement of prefrontal and posterior parietal cortexes in regulating information processing. We conducted behavioral and fMRI experiments to investigate the relationship between memory selection and proactive interference (PI), using a delayed recognition task with a selection cue presented during the delay indicating which two of the four studied digits were relevant to the present test. PI was indexed by the response time differences between rejecting probes matching and not matching the no longer relevant digits. By varying the delay intervals, we found that the effect of PI did not diminish, even for cases in which the postcue interval was extended to 9 sec, but was stronger when the precue interval was lengthened to 5 sec. By examining the correlation between PI index and neural correlates of memory selection, we found that stronger PI is predicted by lower selection-related activity in the left inferior parietal lobe, the precuneus, and the dorsal middle frontal gyrus. Our results suggest that activity in the prefrontal-parietal network may contribute to one's ability to focus on the task-relevant information and may proactively reduce PI in working memory. PMID- 19679762 TI - Auditory context effects in picture naming investigated with event-related fMRI. AB - Naming an object entails a number of processing stages, including retrieval of a target lexical concept and encoding of its phonological word form. We investigated these stages using the picture-word interference task in an fMRI experiment. Participants named target pictures in the presence of auditorily presented semantically related, phonologically related, or unrelated distractor words or in isolation. We observed BOLD signal changes in left-hemisphere regions associated with lexical-conceptual and phonological processing, including the mid to-posterior lateral temporal cortex. However, these BOLD responses manifested as signal reductions for all distractor conditions relative to naming alone. Compared with unrelated words, phonologically related distractors showed further signal reductions, whereas only the pars orbitalis of the left inferior frontal cortex showed a selective reduction in response in the semantic condition. We interpret these findings as indicating that the word forms of lexical competitors are phonologically encoded and that competition during lexical selection is reduced by phonologically related distractors. Since the extended nature of auditory presentation requires a large portion of a word to be presented before its meaning is accessed, we attribute the BOLD signal reductions observed for semantically related and unrelated words to lateral inhibition mechanisms engaged after target name selection has occurred, as has been proposed in some production models. PMID- 19679764 TI - Primed picture naming within and across languages: an ERP investigation. AB - In two experiments, while event-related potentials (ERPs) were recorded, participants named picture targets that were preceded by masked word primes that corresponded either to the name of the picture target or to an unrelated picture name. Experiment 1 showed significant priming effects in the ERP waveforms, free from articulator artifact, starting as early as 200 msec post target onset. Possible loci of these priming effects were proposed within the framework of generic interactive activation models of word recognition and picture naming. These were grouped into three main components: object-specific structural representations, amodal semantic representations, and word-specific phonological and articulatory representations. Experiment 2 provided an initial test of the possible role of each of these components by comparing within-language repetition priming with priming from translation equivalents in bilingual participants. The early and widespread effects of noncognate translation primes in L1 on picture naming in L2 point to object-specific and amodal semantic representations as the principal loci of priming effects obtained with masked word primes and picture targets. PMID- 19679763 TI - Neural correlates of arithmetic calculation strategies. AB - Recent research into math cognition has identified areas of the brain that are involved in number processing (Dehaene, Piazza, Pinel, & Cohen, 2003) and complex problem solving (Anderson, 2007). Much of this research assumes that participants use a single strategy; yet, behavioral research finds that people use a variety of strategies (LeFevre et al., 1996; Siegler, 1987; Siegler & Lemaire, 1997). In the present study, we examined cortical activation as a function of two different calculation strategies for mentally solving multidigit multiplication problems. The school strategy, equivalent to long multiplication, involves working from right to left. The expert strategy, used by "lightning" mental calculators (Staszewski, 1988), proceeds from left to right. The two strategies require essentially the same calculations, but have different working memory demands (the school strategy incurs greater demands). The school strategy produced significantly greater early activity in areas involved in attentional aspects of number processing (posterior superior parietal lobule, PSPL) and mental representation (posterior parietal cortex, PPC), but not in a numerical magnitude area (horizontal intraparietal sulcus, HIPS) or a semantic memory retrieval area (lateral inferior prefrontal cortex, LIPFC). An ACT-R model of the task successfully predicted BOLD responses in PPC and LIPFC, as well as in PSPL and HIPS. PMID- 19679765 TI - Modeling the categorical perception of speech sounds: a step toward biological plausibility. AB - Our native language has a lifelong effect on how we perceive speech sounds. Behaviorally, this is manifested as categorical perception, but the neural mechanisms underlying this phenomenon are still unknown. Here, we constructed a computational model of categorical perception, following principles consistent with infant speech learning. A self-organizing network was exposed to a statistical distribution of speech input presented as neural activity patterns of the auditory periphery, resembling the way sound arrives to the human brain. In the resulting neural map, categorical perception emerges from most single neurons of the model being maximally activated by prototypical speech sounds, while the largest variability in activity is produced at category boundaries. Consequently, regions in the vicinity of prototypes become perceptually compressed, and regions at category boundaries become expanded. Thus, the present study offers a unifying framework for explaining the neural basis of the warping of perceptual space associated with categorical perception. PMID- 19679766 TI - Probing the attentional control theory in social anxiety: an emotional saccade task. AB - Volitional attentional control has been found to rely on prefrontal neuronal circuits. According to the attentional control theory of anxiety, impairment in the volitional control of attention is a prominent feature in anxiety disorders. The present study investigated this assumption in socially anxious individuals using an emotional saccade task with facial expressions (happy, angry, fearful, sad, neutral). The gaze behavior of participants was recorded during the emotional saccade task, in which participants performed either pro- or antisaccades in response to peripherally presented facial expressions. The results show that socially anxious persons have difficulties in inhibiting themselves to reflexively attend to facial expressions: They made more erratic prosaccades to all facial expressions when an antisaccade was required. Thus, these findings indicate impaired attentional control in social anxiety. Overall, the present study shows a deficit of socially anxious individuals in attentional control-for example, in inhibiting the reflexive orienting to neutral as well as to emotional facial expressions. This result may be due to a dysfunction in the prefrontal areas being involved in attentional control. PMID- 19679768 TI - An investigation of auditory contagious yawning. AB - Despite a widespread familiarity with the often compelling urge to yawn after perceiving someone else yawn, an understanding of the neural mechanism underlying contagious yawning remains incomplete. In the present auditory fMRI study, listeners used a 4-point scale to indicate how much they felt like yawning following the presentation of a yawn, breath, or scrambled yawn sound. Not only were yawn sounds given significantly higher ratings, a trait positively correlated with each individual's empathy measure, but relative to control stimuli, random effects analyses revealed enhanced hemodynamic activity in the right posterior inferior frontal gyrus (pIFG) in response to hearing yawns. Moreover, pIFG activity was greatest for yawn stimuli associated with high as opposed to low yawn ratings and for control sounds associated with equally high yawn ratings. These results support a relationship between contagious yawning and empathy and provide evidence for pIFG involvement in contagious yawning. A supplemental figure for this study may be downloaded from http://cabn.psychonomic journals.org/content/supplemental. PMID- 19679767 TI - An electrophysiological investigation into the automaticity of emotional face processing in high versus low trait anxious individuals. AB - To examine the extent of automaticity of emotional face processing in high versus low trait anxious participants, event-related potentials (ERPs) were recorded to emotional (fearful, happy) and neutral faces under varying task demands (low load, high load). Results showed that perceptual encoding of emotional faces, as reflected in P1 and early posterior negativity components, was unaffected by the availability of processing resources. In contrast, the postperceptual registration and storage of emotion-related information, as reflected in the late positive potential component at frontal locations, was influenced by the availability of processing resources, and this effect was further modulated by level of trait anxiety. Specifically, frontal ERP augmentations to emotional faces were eliminated in the more demanding task for low trait anxious participants, whereas ERP enhancements to emotional faces were unaffected by task load in high trait anxious participants. This result suggests greater automaticity in processing affective information in high trait anxious participants. PMID- 19679769 TI - Formation of ArF from LPdAr(F): catalytic conversion of aryl triflates to aryl fluorides. AB - Despite increasing pharmaceutical importance, fluorinated aromatic organic molecules remain difficult to synthesize. Present methods require either harsh reaction conditions or highly specialized reagents, making the preparation of complex fluoroarenes challenging. Thus, the development of general methods for their preparation that overcome the limitations of those techniques currently in use is of great interest. We have prepared [LPd(II)Ar(F)] complexes, where L is a biaryl monophosphine ligand and Ar is an aryl group, and identified conditions under which reductive elimination occurs to form an Ar-F bond. On the basis of these results, we have developed a catalytic process that converts aryl bromides and aryl triflates into the corresponding fluorinated arenes by using simple fluoride salts. We expect this method to allow the introduction of fluorine atoms into advanced, highly functionalized intermediates. PMID- 19679770 TI - High-detectivity polymer photodetectors with spectral response from 300 nm to 1450 nm. AB - Sensing from the ultraviolet-visible to the infrared is critical for a variety of industrial and scientific applications. Today, gallium nitride-, silicon-, and indium gallium arsenide--based detectors are used for different sub-bands within the ultraviolet to near-infrared wavelength range. We demonstrate polymer photodetectors with broad spectral response (300 to 1450 nanometers) fabricated by using a small-band-gap semiconducting polymer blended with a fullerene derivative. Operating at room temperature, the polymer photodetectors exhibit detectivities greater than 10(12) cm Hz(1/2)/W and a linear dynamic range over 100 decibels. The self-assembled nanomorphology and device architecture result in high photodetectivity over this wide spectral range and reduce the dark current (and noise) to values well below dark currents obtained in narrow-band photodetectors made with inorganic semiconductors. PMID- 19679771 TI - An acidic matrix protein, Pif, is a key macromolecule for nacre formation. AB - The mollusk shell is a hard tissue consisting of calcium carbonate crystals and an organic matrix. The nacre of the shell is characterized by a stacked compartment structure with a uniformly oriented c axis of aragonite crystals in each compartment. Using a calcium carbonate-binding assay, we identified an acidic matrix protein, Pif, in the pearl oyster Pinctada fucata that specifically binds to aragonite crystals. The Pif complementary DNA (cDNA) encoded a precursor protein, which was posttranslationally cleaved to produce Pif 97 and Pif 80. The results from immunolocalization, a knockdown experiment that used RNA interference, and in vitro calcium carbonate crystallization studies strongly indicate that Pif regulates nacre formation. PMID- 19679772 TI - Biochemistry. The molecular basis of nacre formation. PMID- 19679774 TI - New land reform in Nigeria. PMID- 19679773 TI - Evidence for obliquity forcing of glacial Termination II. AB - Variations in the intensity of high-latitude Northern Hemisphere summer insolation, driven largely by precession of the equinoxes, are widely thought to control the timing of Late Pleistocene glacial terminations. However, recently it has been suggested that changes in Earth's obliquity may be a more important mechanism. We present a new speleothem-based North Atlantic marine chronology that shows that the penultimate glacial termination (Termination II) commenced 141,000 +/- 2500 years before the present, too early to be explained by Northern Hemisphere summer insolation but consistent with changes in Earth's obliquity. Our record reveals that Terminations I and II are separated by three obliquity cycles and that they started at near-identical obliquity phases. PMID- 19679775 TI - Hydrology. Northern India's groundwater is going, going, going ... PMID- 19679776 TI - Regulatory policy. U.S. panel urges clearer, cleaner role for science. PMID- 19679777 TI - Particle physics. Running at half-energy keeps LHC in race for discoveries. PMID- 19679779 TI - U.S. science policy. Chu lays out an agenda for PCAST and asks for help. PMID- 19679780 TI - Newsmaker interview. Margaret Hamburg aims to strengthen FDA science. Interview by Robert Koenig. PMID- 19679782 TI - Stem cell research. Recipe for induced pluripotent stem cells just got clearer. PMID- 19679783 TI - Energy efficiency. Leaping the efficiency gap. PMID- 19679784 TI - Energy efficiency. Many more more-efficient computers. PMID- 19679785 TI - Energy efficiency. Soap operas to save energy. PMID- 19679786 TI - Energy efficiency. Wanted: help with building design. PMID- 19679787 TI - Energy efficiency. The quest for white LEDs hits the home stretch. PMID- 19679788 TI - Energy efficiency. Aircraft designers shoot for savings on the wing. PMID- 19679789 TI - Energy efficiency. Making use of excess heat. PMID- 19679791 TI - Energy strategies and efficiency. PMID- 19679792 TI - Evolution of the monkey crouch. PMID- 19679793 TI - Plagiarism: consider the context. PMID- 19679794 TI - Plagiarism: transparency required. PMID- 19679795 TI - Defining language boundaries. PMID- 19679796 TI - No paradox for invasive plants. PMID- 19679797 TI - Clarifying coals. PMID- 19679798 TI - Research ethics. Children and population biobanks. PMID- 19679799 TI - Paleontology. Fire and stone. PMID- 19679800 TI - Astronomy. Gamma-ray pulsars old and new. PMID- 19679801 TI - Engineering. Cellulosic biofuels--got gasoline? PMID- 19679802 TI - Behavior. Monkeys like mimics. PMID- 19679803 TI - Physiology. How much sleep do we need? PMID- 19679804 TI - Biochemistry. Friction in motor proteins. PMID- 19679805 TI - Strategic reading, ontologies, and the future of scientific publishing. AB - The revolution in scientific publishing that has been promised since the 1980s is about to take place. Scientists have always read strategically, working with many articles simultaneously to search, filter, scan, link, annotate, and analyze fragments of content. An observed recent increase in strategic reading in the online environment will soon be further intensified by two current trends: (i) the widespread use of digital indexing, retrieval, and navigation resources and (ii) the emergence within many scientific disciplines of interoperable ontologies. Accelerated and enhanced by reading tools that take advantage of ontologies, reading practices will become even more rapid and indirect, transforming the ways in which scientists engage the literature and shaping the evolution of scientific publishing. PMID- 19679806 TI - Local adaptation of bacteriophages to their bacterial hosts in soil. AB - Microbes are incredibly abundant and diverse and are key to ecosystem functioning, yet relatively little is known about the ecological and evolutionary mechanisms that shape their distributions. Bacteriophages, viral parasites that lyse their bacterial hosts, exert intense and spatially varying selection pressures on bacteria and vice versa. We measured local adaptation of bacteria and their associated phages in a centimeter-scale soil population. We first demonstrate that a large proportion of bacteria is sensitive to locally occurring phages. We then show that sympatric phages (isolated from the same 2-gram soil samples as the bacteria) are more infective than are phages from samples some distance away. This study demonstrates the importance of biotic interactions for the small-scale spatial structuring of microbial genetic diversity in soil. PMID- 19679808 TI - Impact of anode microstructure on solid oxide fuel cells. AB - We report a correlation between the microstructure of the anode electrode of a solid oxide fuel cell (SOFC) and its electrochemical performance for a tubular design. It was shown that the electrochemical performance of the cell was extensively improved when the size of constituent particles was reduced so as to yield a highly porous microstructure. The SOFC had a power density of greater than 1 watt per square centimeter at an operating temperature as low as 600 degrees C with a conventional zirconia-based electrolyte, a nickel cermet anode, and a lanthanum ferrite perovskite cathode material. The effect of the hydrogen fuel flow rate (linear velocity) was also examined for the optimization of operating conditions. Higher linear fuel velocity led to better cell performance for the cell with higher anode porosity. A zirconia-based cell could be used for a low-temperature SOFC system under 600 degrees C just by optimizing the microstructure of the anode electrode and operating conditions. PMID- 19679809 TI - Docking in metal-organic frameworks. AB - The use of metal-organic frameworks (MOFs) so far has largely relied on nonspecific binding interactions to host small molecular guests. We used long organic struts (approximately 2 nanometers) incorporating 34- and 36-membered macrocyclic polyethers as recognition modules in the construction of several crystalline primitive cubic frameworks that engage in specific binding in a way not observed in passive, open reticulated geometries. MOF-1001 is capable of docking paraquat dication (PQT2+) guests within the macrocycles in a stereoelectronically controlled fashion. This act of specific complexation yields quantitatively the corresponding MOF-1001 pseudorotaxanes, as confirmed by x-ray diffraction and by solid- and solution-state nuclear magnetic resonance spectroscopic studies performed on MOF-1001, its pseudorotaxanes, and their molecular strut precursors. A control experiment involving the attempted inclusion of PQT2+ inside a framework (MOF-177) devoid of polyether struts showed negligible uptake of PQT2+, indicating the importance of the macrocyclic polyether in PQT2+ docking. PMID- 19679807 TI - Detection of high-energy gamma-ray emission from the globular cluster 47 Tucanae with Fermi. AB - We report the detection of gamma-ray emissions above 200 megaelectron volts at a significance level of 17sigma from the globular cluster 47 Tucanae, using data obtained with the Large Area Telescope onboard the Fermi Gamma-ray Space Telescope. Globular clusters are expected to emit gamma rays because of the large populations of millisecond pulsars that they contain. The spectral shape of 47 Tucanae is consistent with gamma-ray emission from a population of millisecond pulsars. The observed gamma-ray luminosity implies an upper limit of 60 millisecond pulsars present in 47 Tucanae. PMID- 19679810 TI - Fire as an engineering tool of early modern humans. AB - The controlled use of fire was a breakthrough adaptation in human evolution. It first provided heat and light and later allowed the physical properties of materials to be manipulated for the production of ceramics and metals. The analysis of tools at multiple sites shows that the source stone materials were systematically manipulated with fire to improve their flaking properties. Heat treatment predominates among silcrete tools at approximately 72 thousand years ago (ka) and appears as early as 164 ka at Pinnacle Point, on the south coast of South Africa. Heat treatment demands a sophisticated knowledge of fire and an elevated cognitive ability and appears at roughly the same time as widespread evidence for symbolic behavior. PMID- 19679811 TI - Mesotocin and nonapeptide receptors promote estrildid flocking behavior. AB - Proximate neural mechanisms that influence preferences for groups of a given size are almost wholly unknown. In the highly gregarious zebra finch (Estrildidae: Taeniopygia guttata), blockade of nonapeptide receptors by an oxytocin (OT) antagonist significantly reduced time spent with large groups and familiar social partners independent of time spent in social contact. Opposing effects were produced by central infusions of mesotocin (MT, avian homolog of OT). Most drug effects appeared to be female-specific. Across five estrildid finch species, species-typical group size correlates with nonapeptide receptor distributions in the lateral septum, and sociality in female zebra finches was reduced by OT antagonist infusions into the septum but not a control area. We propose that titration of sociality by MT represents a phylogenetically deep framework for the evolution of OT's female-specific roles in pair bonding and maternal functions. PMID- 19679812 TI - The transcriptional repressor DEC2 regulates sleep length in mammals. AB - Sleep deprivation can impair human health and performance. Habitual total sleep time and homeostatic sleep response to sleep deprivation are quantitative traits in humans. Genetic loci for these traits have been identified in model organisms, but none of these potential animal models have a corresponding human genotype and phenotype. We have identified a mutation in a transcriptional repressor (hDEC2 P385R) that is associated with a human short sleep phenotype. Activity profiles and sleep recordings of transgenic mice carrying this mutation showed increased vigilance time and less sleep time than control mice in a zeitgeber time- and sleep deprivation-dependent manner. These mice represent a model of human sleep homeostasis that provides an opportunity to probe the effect of sleep on human physical and mental health. PMID- 19679813 TI - Protein friction limits diffusive and directed movements of kinesin motors on microtubules. AB - Friction limits the operation of macroscopic engines and is critical to the performance of micromechanical devices. We report measurements of friction in a biological nanomachine. Using optical tweezers, we characterized the frictional drag force of individual kinesin-8 motor proteins interacting with their microtubule tracks. At low speeds and with no energy source, the frictional drag was related to the diffusion coefficient by the Einstein relation. At higher speeds, the frictional drag force increased nonlinearly, consistent with the motor jumping 8 nanometers between adjacent tubulin dimers along the microtubule, and was asymmetric, reflecting the structural polarity of the microtubule. We argue that these frictional forces arise from breaking bonds between the motor domains and the microtubule, and they limit the speed and efficiency of kinesin. PMID- 19679814 TI - Mechanistic analysis of a dynamin effector. AB - Dynamin-related proteins (DRPs) can generate forces to remodel membranes. In cells, DRPs require additional proteins [DRP-associated proteins (DAPs)] to conduct their functions. To dissect the mechanistic role of a DAP, we used the yeast mitochondrial division machine as a model, which requires the DRP Dnm1, and two other proteins, Mdv1 and Fis1. Mdv1 played a postmitochondrial targeting role in division by specifically interacting and coassembling with the guanosine triphosphate-bound form of Dnm1. This regulated interaction nucleated and promoted the self-assembly of Dnm1 into helical structures, which drive membrane scission. The nucleation of DRP assembly probably represents a general regulatory strategy for this family of filament-forming proteins, similar to F-actin regulation. PMID- 19679815 TI - ER stress controls iron metabolism through induction of hepcidin. AB - Hepcidin is a peptide hormone that is secreted by the liver and controls body iron homeostasis. Hepcidin overproduction causes anemia of inflammation, whereas its deficiency leads to hemochromatosis. Inflammation and iron are known extracellular stimuli for hepcidin expression. We found that endoplasmic reticulum (ER) stress also induces hepcidin expression and causes hypoferremia and spleen iron sequestration in mice. CREBH (cyclic AMP response element-binding protein H), an ER stress-activated transcription factor, binds to and transactivates the hepcidin promoter. Hepcidin induction in response to exogenously administered toxins or accumulation of unfolded protein in the ER is defective in CREBH knockout mice, indicating a role for CREBH in ER stress regulated hepcidin expression. The regulation of hepcidin by ER stress links the intracellular response involved in protein quality control to innate immunity and iron homeostasis. PMID- 19679816 TI - Capuchin monkeys display affiliation toward humans who imitate them. AB - During social interactions, humans often unconsciously and unintentionally imitate the behaviors of others, which increases rapport, liking, and empathy between interaction partners. This effect is thought to be an evolutionary adaptation that facilitates group living and may be shared with other primate species. Here, we show that capuchin monkeys, a highly social primate species, prefer human imitators over non-imitators in a variety of ways: The monkeys look longer at imitators, spend more time in proximity to imitators, and choose to interact more frequently with imitators in a token exchange task. These results demonstrate that imitation can promote affiliation in nonhuman primates. Behavior matching that leads to prosocial behaviors toward others may have been one of the mechanisms at the basis of altruistic behavioral tendencies in capuchins and in other primates, including humans. PMID- 19679818 TI - P2Y5 is a G(alpha)i, G(alpha)12/13 G protein-coupled receptor activated by lysophosphatidic acid that reduces intestinal cell adhesion. AB - P2Y5 is a G protein-coupled receptor that binds and is activated by lysophosphatidic acid (LPA). We determined that P2Y5 transcript is expressed along the intestinal mucosa and investigated the intracellular pathways induced by P2Y5 activation, which could contribute to LPA effects on intestinal cell adhesion. P2Y5 heterologously expressed in CHO and small intestinal hBRIE 380i cells was activated by LPA resulting in an increase in intracellular calcium ([Ca(2+)](i)) when the cells concurrently expressed G(alpha)(Delta6qi5myr). P2Y5 activation also increased the phosphorylation of ERK1/2 that was sensitive to pertussis toxin. Together these indicate that P2Y5 activation by LPA induces an increase in [Ca(2+)](i) and ERK1/2 phosphorylation through G(alpha)(i). We discovered that P2Y5 was activated by farnesyl pyrophosphate (FPP) without a detectable change in [Ca(2+)](i). The activation of P2Y5 by LPA or FPP induced the activity of a serum response element (SRE)-linked luciferase reporter that was inhibited by the RGS domain of p115RhoGEF, C3 exotoxin, and Y-27632, suggesting the involvement of G(alpha)(12/13), Rho GTPase, and ROCK, respectively. However, only LPA-mediated induction of SRE reporter activity was sensitive to inhibitors targeting p38 MAPK, PI3K, PLC, and PKC. In addition, only LPA transactivated the epidermal growth factor receptor, leading to an induction of ERK1/2 phosphorylation. These observations correlate with our subsequent finding that P2Y5 activation by LPA, and not FPP, reduced intestinal cell adhesion. This study elucidates a mechanism whereby LPA can act as a luminal and/or serosal cue to alter mucosal integrity. PMID- 19679819 TI - Role of PSD95 in membrane association and catalytic activity of nNOSalpha in nitrergic varicosities in mice gut. AB - We have recently shown that membrane association of neuronal nitric oxide synthase-alpha (nNOSalpha) is critical in the regulation of synthesis of NO during nitrergic neurotransmission. The purpose of this study was to examine the role of the synapse-associated proteins (SAPs) in membrane association of nNOSalpha. Varicosities (swellings on terminal axons) were isolated from mice gastrointestinal tract and examined for nNOSalpha, postsynaptic density protein 95 (PSD95), and membrane interactions by coimmunoprecipitation and SDS-PAGE. Our results show that PSD95 protein was present in the membrane fraction of the nerve varicosity, whereas both PSD95 and SAP97 were present in the cytosol. nNOSalpha was associated with PSD95 but not SAP97. nNOSalpha-PSD95 complex was bound to the membrane via palmitoylation of PSD95. Depalmitoylation of PSD95 with 2 bromopalmitate dislocates nNOSalpha and PSD95 from the varicosity membrane and abolishes NO production. These studies show that palmitoylation of PSD95 anchors nNOSalpha to the varicosity membrane and that it is obligatory for NO production by the enzyme. Palmitoylation of PSD95 may provide a novel target for regulation of nitrergic neurotransmission. PMID- 19679820 TI - Hepatic uptake of gamma-butyrobetaine, a precursor of carnitine biosynthesis, in rats. AB - Gamma-butyrobetaine (GBB) is a precursor in the biosynthesis of carnitine, which plays an important role in the beta-oxidation of fatty acids, and is converted to carnitine by gamma-butyrobetaine dioxygenase (BBD) predominantly in liver. We investigated the molecular mechanism of hepatic uptake of GBB in rat hepatocytes. Cellular localization of rat Octn2 (rOctn2:Slc22A5) was studied by Western blot analysis. Uptake of deuterated GBB (d(3)-GBB) was examined in HEK293 cells expressing rOctn2 (HEK293/rOctn2) and freshly isolated rat hepatocytes. d(3)-GBB was quantified by use of liquid chromatography-tandem mass spectrometry. Western blot analysis demonstrated an expression of OCTN2 protein in hepatic basolateral membrane but not in bile canalicular membrane fraction. Furthermore, we found that d(3)-GBB was taken up by rOctn2 in an Na(+)-dependent manner with K(m) value of 13 microM. The apparent K(m) value for d(3)-GBB transport in freshly isolated rat hepatocytes was 9 microM. d(3)-GBB uptake by the rat hepatocytes was inhibited by gamma-aminobutyric acid (GABA) to 30% of the control, whereas it was inhibited by carnitine to 62% of the control, even at 500 microM. Furthermore, d(3)-GBB uptake by rat hepatocytes was decreased by 45% with rat Gat2 (Slc6A13, a major liver GABA transporter) silenced by the microRNA method. Accordingly, the present study clearly demonstrates that GBB is taken up by hepatocytes for carnitine biosynthesis not only via Octn2 but also via the GABA transporter, possibly Gat2. PMID- 19679821 TI - Alternative splicing of the Menkes copper Atpase (Atp7a) transcript in the rat intestinal epithelium. AB - The intestinal Menkes copper Atpase (Atp7a) gene is strongly induced by iron deficiency in the rat intestine. We sought to develop an in vitro model to understand the mechanism of this induction by performing molecular studies in native rat intestine and in intestinal epithelial (IEC-6) cells. IEC-6 cells express Atp7a, and induction was noted with iron deprivation. 5' Rapid amplification of cDNA ends and PCR experiments revealed three splice variants in rat intestine and IEC-6 cells; all variants were strongly induced during iron deprivation (five- to sevenfold). The splice variants presumably encode proteins that would either contain the extreme NH(2) terminus of the protein (containing copper binding domain 1) or not. We thus hypothesized that more than one version of Atp7a protein exists. Antibodies against this NH(2)-terminal region of the protein were developed (named N-term) and used along with previously reported antibodies (against more COOH-terminal regions, termed 54-10) to perform immunoblotting and immunolocalization studies. Results with the 54-10 antiserum revealed an Atp7a protein variant of approximately 190 kDa that localized to the trans-Golgi network of IEC-6 cells and trafficked to the plasma membrane with copper loading. Using the N-term antiserum, however, we noted protein of approximately 97 and 64 kDa. The 97-kDa protein was cytosolic and nuclear, whereas the 64-kDa protein was nuclear specific. Immunolocalization analyses with the N-term antiserum showed strong staining of nuclei in IEC-6 and Caco-2 cells and in rat intestine. We conclude that novel Atp7a protein variants may exist in rat and human intestinal epithelial cells, with different intracellular locations and potentially distinct physiological functions. PMID- 19679822 TI - Endocytic trafficking from the small intestinal brush border probed with FM dye. AB - The small intestinal brush border functions as the body's main portal for uptake of dietary nutrients and simultaneously acts as the largest permeability barrier against pathogens. To enable this, the digestive enzymes of the brush border are organized in lipid raft microdomains stabilized by cross-linking galectins and intelectin, but little is known about the dynamic properties of this highly specialized membrane. Here, we probed the endocytic membrane trafficking from the brush border of organ-cultured pig intestinal mucosal explants by use of a fixable, lipophilic FM dye. The fluorescent dye readily incorporated into the brush border, and by 15 min faint but distinct punctae were detectable approximately 1 microm beneath the brush border, indicative of a constitutive endocytosis. The punctae represented a subpopulation of early endosomes confined to the actomyosin-rich terminal web region, and their number and intensity increased by 1 h, but trafficking further into the enterocyte was not observed except in immature epithelial cells of the crypts. A powerful ligand for receptor mediated endocytosis, cholera toxin B subunit, increased apical endocytosis and caused membrane trafficking to proceed to compartments localized deeper into the cytoplasm of the enterocytes. Two major raft-associated brush border enzymes, alkaline phosphatase and aminopeptidase N, were excluded from endocytosis. We propose that the terminal web cytoskeleton, by inhibiting traffic from apical early endosomes further into the cell, contributes to the overall permeability barrier of the gut. PMID- 19679823 TI - Extrinsic afferent nerve sensitivity and enteric neurotransmission in murine jejunum in vitro. AB - Enteric and extrinsic sensory neurons respond to similar stimuli. Thus they may be activated in series or in parallel. Because signal transmission via synapses or mediator release would depend on calcium, we investigated its role for extrinsic afferent sensitivity to chemical and mechanical stimulation. Extracellular multiunit afferent recordings were made in vitro from paravascular nerve bundles supplying the mouse jejunum. Intraluminal pressure and afferent nerve responses were recorded under control conditions and under four conditions designed to interfere with enteric neurotransmission. We found that phasic intestinal contractions ceased after switching perfusion to Ca(2+)-free buffer with or without a purinergic P2 receptor antagonist, pyridoxal phosphate-6 azo(benzene-2,4-disulfonic acid) (PPADS) or cadmium (blocking all Ca(2+) channels) but not following omega-conotoxin GVIA (N-type Ca(2+)-channel blocker). Luminal HCl (pH 2) and 5-HT (500 microM) evoked peak firing of 17 +/- 4 impulses per second (imp/s) (n = 10) and 21 +/- 4 imp/s (n = 13) under control conditions. These responses were reduced to 4 +/- 2 imp/s and 5 +/- 2 imp/s by cadmium (n = 7, P < 0.05), to 7 +/- 2 imp/s and 6 +/- 1 imp/s by Ca(2+)-free perfusion (n = 6, P < 0.05), and to 3 +/- 1 imp/s and 4 +/- 1 imp/s by Ca(2+)-free perfusion with PPADS (n = 6, P < 0.05). Responses were unchanged by omega-conotoxin GVIA. Mechanical ramp distension of the intestinal segment to 60 cmH(2)O was not altered by any of the experimental conditions. We concluded that HCl and 5-HT activate extrinsic afferents via a calcium-dependent mechanism, which is unlikely to involve enteric neurons carrying N-type calcium channels. Extrinsic mechanosensitivity is independent of enteric neurotransmission. It appears that cross talk from the enteric to the extrinsic nervous system does not mediate extrinsic afferent sensitivity. PMID- 19679825 TI - Stability and aggregation of ranked gene lists. AB - Ranked gene lists are highly instable in the sense that similar measures of differential gene expression may yield very different rankings, and that a small change of the data set usually affects the obtained gene list considerably. Stability issues have long been under-considered in the literature, but they have grown to a hot topic in the last few years, perhaps as a consequence of the increasing skepticism on the reproducibility and clinical applicability of molecular research findings. In this article, we review existing approaches for the assessment of stability of ranked gene lists and the related problem of aggregation, give some practical recommendations, and warn against potential misuse of these methods. This overview is illustrated through an application to a recent leukemia data set using the freely available Bioconductor package GeneSelector. PMID- 19679824 TI - Three epigenetic drugs up-regulate homeobox gene Rhox5 in cancer cells through overlapping and distinct molecular mechanisms. AB - Epigenetic therapy of cancer using inhibitors of DNA methyltransferases (DNMT) or/and histone deacetylases (HDACs) has shown promising results in preclinical models and is being investigated in clinical trials. Homeodomain proteins play important roles in normal development and carcinogenesis. In this study, we demonstrated for the first time that an epigenetic drug could up-regulate homeobox genes in the reproductive homeobox genes on chromosome X (Rhox) family, including murine Rhox5, Rhox6, and Rhox9 and human RhoxF1 and RhoxF2 in breast, colon, and other types of cancer cells. We examined the molecular mechanisms underlining selective induction of Rhox5 in cancer cells by three epigenetic drugs: 5-aza-2'-deoxycytidine (DAC; decitabine), arsenic trioxide (ATO), and MS 275 [entinostat; N-(2-aminophenyl)-4-[N-(pyridine-3-ylmethoxy carbonyl)aminomethyl]benzamide]. DAC induced Rhox5 mRNA expression from both distal promoter (Pd) and proximal promoter, whereas MS-275 and ATO induced gene expression from the Pd only. DAC and ATO inhibited both DNMT1 and DNMT3B protein expression, whereas MS-275 significantly reduced DNMT3B protein. In contrast to DAC, neither MS-275 nor ATO induced DNA demethylation on the Pd region. All three drugs led to enhanced acetylation of histones H3 and H4 at the promoter region. The occupancy of the activating histone mark dimethylated lysine 4 of H3 at Pd was enhanced by DAC and MS-275 but not ATO. Because they modulate gene expression with different potencies through shared and distinct epigenetic mechanisms, these epigenetic drugs may possess great potential in different applications for epigenetic therapy of cancer and other diseases. PMID- 19679826 TI - Inhibition of long-chain acyl coenzyme A synthetases during fatty acid loading induces lipotoxicity in macrophages. AB - OBJECTIVE: Obesity is often associated with hypertriglyceridemia and elevated free fatty acids (FFAs), which are independent risk factors for cardiovascular disease and diabetes. Although impairment of cholesterol homeostasis is known to induce toxicity in macrophages, the consequence of altered fatty acid homeostasis is not clear. METHODS AND RESULTS: Long-chain acyl CoA synthetases (ACSLs) play a critical role in fatty acid homeostasis by channeling fatty acids to diverse metabolic pools. We treated mouse peritoneal macrophages (MPMs) with VLDL or FFAs in the presence of triacsin C, an inhibitor of the 3 ACSL isoforms present in macrophages. Treatment of macrophages with VLDL and triacsin C resulted in reduced TG accumulation but increased intracellular FFA levels, which induced lipotoxicity characterized by apoptosis. Treatment of MPMs with the saturated fatty acid stearic acid in the presence of triacsin C increased intracellular stearic acid and induced apoptosis. Stromal vascular cells collected from high fat diet-fed mice displayed foam cell morphology and exhibited increased mRNA levels of macrophage markers and ACSL1. Importantly, all of these changes were associated with increased FFA level in AT. CONCLUSIONS: Inhibition of ACSLs during fatty acid loading results in apoptosis via accumulation of FFAs. Our data have implications in understanding the consequences of dysregulated fatty acid metabolism in macrophages. PMID- 19679827 TI - Statins block calcific nodule formation of valvular interstitial cells by inhibiting alpha-smooth muscle actin expression. AB - OBJECTIVE: Calcific aortic stenosis, characterized by excessive fibrosis and deposition of bone-like calcified tissue, affects roughly 2% to 3% of the U.S. population over the age of 65. Recent studies have suggested that statins have a positive effect on the progression of aoritic stenosis, likely because of their ability to affect the resident cell population, known as valvular interstitial cells (VICs). VICs are fibroblastic cells that can differentiate to form activated myofibroblasts, displaying increased alpha smooth muscle actin (alphaSMA) expression, contractility, and collagen production. METHODS AND RESULTS: In culture, VICs spontaneously form multicellular aggregates that subsequently develop into calcified nodules, providing an in vitro model for aortic stenosis. Using real-time microscopic tracking, we observed that confluent VIC monolayers spontaneously contract into rounded nodules, suggesting that myofibroblastic contractility is a critical step in the process of nodule formation. Overexpression of alphaSMA increased VIC calcific nodule formation and contractility, whereas knockdown of alphaSMA with siRNAs reduced these phenotypes, suggesting that the expression and contractile properties of alphaSMA are essential to the formation of nodules. Statin treatment of VICs reduced alphaSMA expression, inhibited contractility, and decreased nodule formation. When statins were used to treat preformed nodules, no decrease in the number of calcified nodules was observed, suggesting that statins may play more of a preventative role in aortic stenosis than a cure. CONCLUSIONS: Our studies provide evidence of a causal relationship between VIC myofibroblastic activity and initial VIC calcific nodule formation. Furthermore, we demonstrate that pravastatin inhibition of calcific nodule formation is related to inhibition of myofibroblastic activity. PMID- 19679828 TI - Liver X receptor-mediated induction of cholesteryl ester transfer protein expression is selectively impaired in inflammatory macrophages. AB - OBJECTIVE: Cholesteryl ester transfer protein (CETP) is a target gene for the liver X receptor (LXR). The aim of this study was to further explore this regulation in the monocyte-macrophage lineage and its modulation by lipid loading and inflammation, which are key steps in the process of atherogenesis. METHODS AND RESULTS: Exposure of bone marrow-derived macrophages from human CETP transgenic mice to the T0901317 LXR agonist increased CETP, PLTP, and ABCA1 mRNA levels. T0901317 also markedly increased CETP mRNA levels and CETP production in human differentiated macrophages, whereas it had no effect on CETP expression in human peripheral blood monocytes. In inflammatory mouse and human macrophages, LXR-mediated CETP gene upregulation was inhibited, even though ABCA1, ABCG1, and SREBP1c inductions were maintained. The inhibition of CETP gene response to LXR agonists in inflammatory cells was independent of lipid loading (ie, oxidized LDL increased CETP production in noninflammatory macrophages with a synergistic effect of synthetic LXR agonists). CONCLUSIONS: LXR-mediated induction of human CETP expression is switched on during monocyte-to-macrophage differentiation, is magnified by lipid loading, and is selectively lost in inflammatory macrophages, which suggests that inflammatory cells may not increase the circulating CETP pool on LXR agonist treatment. PMID- 19679829 TI - Lack of tyrosylprotein sulfotransferase activity in hematopoietic cells drastically attenuates atherosclerosis in Ldlr-/- mice. AB - OBJECTIVE: Leukocyte recruitment is a major contributor in the development of atherosclerosis and requires a variety of proteins such as adhesion molecules, chemokines, and chemokine receptors. Several key molecular players implicated in this process are expressed on monocytes and require protein-tyrosine sulfation for optimal function in vitro, including human CCR2, CCR5, CX3CR1, and PSGL-1. We therefore hypothesized that protein-tyrosine sulfation in hematopoietic cells plays an important role in the development of atherosclerosis. METHODS AND RESULTS: Lethally-irradiated Ldlr(-/-) mice were rescued with hematopoietic progenitors lacking tyrosylprotein sulfotransferase (TPST) activity attributable to deletion of the Tpst1 and Tpst2 genes. TPST deficient progenitors efficiently reconstituted hematopoiesis in Ldlr(-/-) recipients and transplantation had no effect on plasma lipids on a standard or atherogenic diet. However, we observed a substantial reduction in the size of atherosclerotic lesions and the number of macrophages in lesions from hyperlipidemic Ldlr(-/-) recipients transplanted with TPST deficient progenitors compared to wild-type progenitors. We also document for the first time that murine Psgl-1 and Cx3cr1 are tyrosine-sulfated. CONCLUSIONS: These data demonstrate that protein-tyrosine sulfation is an important contributor to monocytes/macrophage recruitment and/or retention in a mouse model of atherosclerosis. PMID- 19679830 TI - Therapeutic potential of unrestricted somatic stem cells isolated from placental cord blood for cardiac repair post myocardial infarction. AB - OBJECTIVE: Unrestricted somatic stem cells (USSCs) were successfully identified from human cord blood. However, the efficacy of USSC transplantation for improving left ventricular (LV) function post myocardial infarction (MI) is still controversial. METHODS AND RESULTS: PBS, 1x10(6) human fibroblasts (Fbr), 1x10(5) USSCs (LD), or 1x10(6) USSCs (HD) were transplanted intramyocardially 20 minutes after ligating the LAD of nude rats. Echocardiography and a microtip conductance catheter at day 28 revealed a dose-dependent improvement of LV function after USSC transplantation. Necropsy examination revealed dose-dependent augmentation of capillary density and inhibition of LV fibrosis. Dual-label immunohistochemistry for cardiac troponin-I and human nuclear antigen (HNA) demonstrated that human cardiomyocytes (CMCs) were dose-dependently generated in ischemic myocardium 28 days after USSC transplantation. Similarly, dual-label immunostaining for smooth muscle actin and class I human leukocyte antigen or that for von Willebrand factor and HNA also revealed a dose-dependent vasculogenesis after USSC transplantation. RT-PCR indicated that expression of human-specific genes of CMCs, smooth muscle cells, and endothelial cell markers in infarcted myocardium were significantly augmented in USSC-treated animals compared with control groups. CONCLUSIONS: USSC transplantation leads to functional improvement and recovery from MI and exhibits a significant and dose dependent potential for concurrent cardiomyogenesis and vasculogenesis. PMID- 19679831 TI - ENPP1 Q121 variant, increased pulse pressure and reduced insulin signaling, and nitric oxide synthase activity in endothelial cells. AB - OBJECTIVE: Insulin resistance induces increased pulse pressure (PP), endothelial dysfunction (ED), and reduced bioavailability of endothelium-derived nitric oxide (NO). The genetic background of these 3 cardiovascular risk factors might be partly common. The ENPP1 K121Q polymorphism is associated with insulin resistance and cardiovascular risk. METHODS AND RESULTS: We investigated whether the K121Q polymorphism is associated with increased PP in white Caucasians and with ED in vitro. In 985 individuals, (390 unrelated and 595 from 248 families), the K121Q polymorphism was associated with PP (P=8.0 x 10(-4)). In the families, the Q121 variant accounted for 0.08 of PP heritability (P=9.4 x 10(-4)). This association was formally replicated in a second sample of 475 individuals (P=2.6 x 10(-2)) but not in 2 smaller samples of 289 and 236 individuals (P=0.49 and 0.21, respectively). In the individual patients' data meta-analysis, comprising 1985 individuals, PP was associated with the Q121 variant (P=1.2 x 10(-3)). Human endothelial cells carrying the KQ genotype showed, as compared to KK cells, reduced insulin-mediated insulin receptor autophosphorylation (P=0.03), Ser(473) Akt phosphorylation (P=0.03), and NO synthase activity (P=0.003). CONCLUSIONS: Our data suggest that the ENPP1 Q121 variant is associated with increased PP in vivo and reduced insulin signaling and ED in vitro, thus indicating a possible pathogenic mechanism for the increased cardiovascular risk observed in ENPP1 Q121 carriers. PMID- 19679832 TI - Dysfunctional HDL as a diagnostic and therapeutic target. AB - The atheroprotective effects of HDL are mediated by several mechanisms, including its role in reverse cholesterol transport and via its antiinflammatory properties. However, not all HDL is functionally similar. HDL and apolipoprotein A-I may become dysfunctional or even proinflammatory and thus promote atherosclerosis. ApoAI posttranslational modification can have a large impact on its function. Myeloperoxidase modification of apoAI impairs its function as a cholesterol acceptor, and the molecular changes induced by myeloperoxidase have been studied in detail. These studies provide the basis for the development of an oxidant-resistant form of apoAI and clinical measures of HDL modification and dysfunction, which may be useful as a treatment criterion. PMID- 19679833 TI - Correlation between carotid intimal/medial thickness and atherosclerosis: a point of view from pathology. AB - A widely adopted surrogate for predicting rates of cardiovascular events involves measure of carotid intimal-medial thickness (CIMT) by B mode ultrasound, a technique available since the mid 1980s. The value of this modality remains in its ability to noninvasively assess cardiovascular risk beyond traditional factors identified by the Framingham risk score, and it is among the few available techniques for monitoring the effectiveness of pharmacotherapy on plaques. There are, however, existing limitations to this methodology. Perhaps the most important distinction is that IM thickness measurements are generally acquired in the common carotid artery, whereas advanced atherosclerotic disease occurs predominantly downstream in the internal carotid. Moreover, primary contributors to IM thickening are age and hypertension, which do not necessarily reflect the atherosclerotic process. Initiation of disease-related plaques begins as what is referred to as pathological intimal thickening; lesions characterized by the formation of lipid pools in the absence of a necrotic core. The eventual development of a necrotic core, however, is considered a key indicator of significant plaque advancement and recognized feature of lesion vulnerability. Necrotic cores are thought to arise from macrophage infiltration of lipid pools followed by secondary necrosis where defective clearance of debris, tissue disruption proteases, and intraplaque hemorrhage, likely contribute to its enlargement. Therefore, one of the primary limitations to CIMT is its inability to distinguish lesions with a necrotic core. Moreover, in most cases measures of plaque area or volume are generally considered better predictors of an inflammatory process consistent with atherosclerotic disease rather than intimal medial thickness. PMID- 19679834 TI - NADPH oxidase Nox2 is required for hypoxia-induced mobilization of endothelial progenitor cells. AB - RATIONALE: Endothelial progenitor cells (EPCs, defined as sca-1(+)flk-1(+)lin(-) mononuclear blood cells) contribute to vascular repair. The role of hypoxia and reactive oxygen species (ROS) in mobilization and function of these cells is incompletely understood. OBJECTIVE: We studied the contribution of the NADPH oxidase Nox2, an important vascular source of ROS in this context. METHODS AND RESULTS: Hypoxia (10% oxygen) induced the mobilization of EPCs in wild-type (WT) and Nox1 but not in Nox2 knockout (Nox2(y/-)) mice. As erythropoietin (EPO) is known to induce EPC mobilization, we focused on this hormone. EPO induced the mobilization of EPCs in WT and Nox1(y/-) but not Nox2(y/-) animals. Transplantation of bone marrow from Nox2(y/-) mice into WT-mice blocked mobilization in response to hypoxia and EPO, whereas transplantation of WT bone marrow into Nox2(y/-) mice restored mobilization. Reendothelialization of the injured mouse carotid artery was enhanced by hypoxia as well as by EPO, and this effect was not observed in Nox2(y/-) mice or after transplantation of Nox2(y/-) bone marrow. In cultured EPCs from WT but not Nox2(y/-) mice, EPO induced ROS production, migration, and proliferation. EPO signaling involves the STAT5 transcription factor. EPO-induced STAT5-dependent reporter gene expression was absent in Nox2-deficient cells. siRNA against the redox-sensitive phosphatase SHP 2 restored EPO-mediated STAT5 induction and inhibition of SHP-2 restored EPO induced migration in Nox2-deficient cells CONCLUSIONS: We conclude that Nox2 derived ROS inactivate SHP-2 and thereby facilitate EPO signaling in EPCs to promote hypoxia-induced mobilization and vascular repair by these cells. PMID- 19679835 TI - Truncation of titin's elastic PEVK region leads to cardiomyopathy with diastolic dysfunction. AB - RATIONALE: The giant protein titin plays key roles in myofilament assembly and determines the passive mechanical properties of the sarcomere. The cardiac titin molecule has 2 mayor elastic elements, the N2B and the PEVK region. Both have been suggested to determine the elastic properties of the heart with loss of function data only available for the N2B region. OBJECTIVE: The purpose of this study was to investigate the contribution of titin's proline-glutamate-valine lysine (PEVK) region to biomechanics and growth of the heart. METHODS AND RESULTS: We removed a portion of the PEVK segment (exons 219 to 225; 282 aa) that corresponds to the PEVK element of N2B titin, the main cardiac titin isoform. Adult homozygous PEVK knockout (KO) mice developed diastolic dysfunction, as determined by pressure-volume loops, echocardiography, isolated heart experiments, and muscle mechanics. Immunoelectron microscopy revealed increased strain of the N2B element, a spring region retained in the PEVK-KO. Interestingly, the PEVK-KO mice had hypertrophied hearts with an induction of the hypertrophy and fetal gene response that includes upregulation of FHL proteins. This contrasts the cardiac atrophy phenotype with decreased FHL2 levels that result from the deletion of the N2B element. CONCLUSIONS: Titin's PEVK region contributes to the elastic properties of the cardiac ventricle. Our findings are consistent with a model in which strain of the N2B spring element and expression of FHL proteins trigger cardiac hypertrophy. These novel findings provide a molecular basis for the future differential therapy of isolated diastolic dysfunction versus more complex cardiomyopathies. PMID- 19679837 TI - Interval training normalizes cardiomyocyte function, diastolic Ca2+ control, and SR Ca2+ release synchronicity in a mouse model of diabetic cardiomyopathy. AB - RATIONALE: In the present study we explored the mechanisms behind excitation contraction (EC) coupling defects in cardiomyocytes from mice with type-2 diabetes (db/db). OBJECTIVE: We determined whether 13 weeks of aerobic interval training could restore cardiomyocyte Ca(2+) cycling and EC coupling. METHODS AND RESULTS: Reduced contractility in cardiomyocytes isolated from sedentary db/db was associated with increased diastolic sarcoplasmic reticulum (SR)-Ca(2+) leak, reduced synchrony of Ca(2+) release, reduced transverse (T)-tubule density, and lower peak systolic and diastolic Ca(2+) and caffeine-induced Ca(2+) release. Additionally, the rate of SR Ca(2+) ATPase-mediated Ca(2+) uptake during diastole was reduced, whereas a faster recovery from caffeine-induced Ca(2+) release indicated increased Na(+)/Ca(2+)-exchanger activity. The increased SR-Ca(2+) leak was attributed to increased Ca(2+)-calmodulin-dependent protein kinase (CaMKIIdelta) phosphorylation, supported by the normalization of SR-Ca(2+) leak on inhibition of CaMKIIdelta (AIP). Exercise training restored contractile function associated with restored SR Ca(2+) release synchronicity, T-tubule density, twitch Ca(2+) amplitude, SR Ca(2+) ATPase and Na(+)/Ca(2+)-exchanger activities, and SR-Ca(2+) leak. The latter was associated with reduced phosphorylation of cytosolic CaMKIIdelta. Despite normal contractile function and Ca(2+) handling after the training period, phospholamban was hyperphosphorylated at Serine-16. Protein kinase A inhibition (H-89) in cardiomyocytes from the exercised db/db group abolished the differences in SR-Ca(2+) load when compared with the sedentary db/db mice. EC coupling changes were observed without changes in serum insulin or glucose levels, suggesting that the exercise training-induced effects are not via normalization of the diabetic condition. CONCLUSIONS: These data demonstrate that aerobic interval training almost completely restored the contractile function of the diabetic cardiomyocyte to levels close to sedentary wild type. PMID- 19679836 TI - Loss of cardiac microRNA-mediated regulation leads to dilated cardiomyopathy and heart failure. AB - RATIONALE: Heart failure is a deadly and devastating disease that places immense costs on an aging society. To develop therapies aimed at rescuing the failing heart, it is important to understand the molecular mechanisms underlying cardiomyocyte structure and function. OBJECTIVE: microRNAs are important regulators of gene expression, and we sought to define the global contributions made by microRNAs toward maintaining cardiomyocyte integrity. METHODS AND RESULTS: First, we performed deep sequencing analysis to catalog the miRNA population in the adult heart. Second, we genetically deleted, in cardiac myocytes, an essential component of the machinery that is required to generate miRNAs. Deep sequencing of miRNAs from the heart revealed the enrichment of a small number of microRNAs with one, miR-1, accounting for 40% of all microRNAs. Cardiomyocyte-specific deletion of dgcr8, a gene required for microRNA biogenesis, revealed a fully penetrant phenotype that begins with left ventricular malfunction progressing to a dilated cardiomyopathy and premature lethality. CONCLUSIONS: These observations reveal a critical role for microRNAs in maintaining cardiac function in mature cardiomyocytes and raise the possibility that only a handful of microRNAs may ultimately be responsible for the dramatic cardiac phenotype seen in the absence of dgcr8. PMID- 19679838 TI - Platelet kainate receptor signaling promotes thrombosis by stimulating cyclooxygenase activation. AB - RATIONALE: Glutamate is a major signaling molecule that binds to glutamate receptors including the ionotropic glutamate receptors; kainate (KA) receptor (KAR), the N-methyl-d-aspartate receptor, and the alpha-amino-3-hydroxy-5-methyl 4-isoxazolepropionic acid receptor. Each is well characterized in the central nervous system, but glutamate has important signaling roles in peripheral tissues as well, including a role in regulating platelet function. OBJECTIVE: Our previous work has demonstrated that glutamate is released by platelets in high concentrations within a developing thrombus and increases platelet activation and thrombosis. We now show that platelets express a functional KAR that drives increased agonist induced platelet activation. METHODS AND RESULTS: KAR induced increase in platelet activation is in part the result of activation of platelet cyclooxygenase in a mitogen-activated protein kinase-dependent manner. Platelets derived from KAR subunit knockout mice (GluR6(-/-)) are resistant to KA effects and have a prolonged time to thrombosis in vivo. Importantly, we have also identified polymorphisms in KAR subunits that are associated with phenotypic changes in platelet function in a large group of whites and blacks. CONCLUSIONS: Our data demonstrate that glutamate regulation of platelet activation is in part cyclooxygenase-dependent and suggest that the KAR is a novel antithrombotic target. PMID- 19679839 TI - PKC phosphorylation of titin's PEVK element: a novel and conserved pathway for modulating myocardial stiffness. AB - RATIONALE: Protein kinase C (PKC) regulates contractility of cardiac muscle cells by phosphorylating thin- and thick- filament-based proteins. Myocardial sarcomeres also contain a third myofilament, titin, and it is unknown whether titin can be phosphorylated by PKC and whether it affects passive tension. OBJECTIVE: The purpose of this study was to examine the effect of PKC on titin phosphorylation and titin-based passive tension. METHODS AND RESULTS: Phosphorylation assays with PKCalpha revealed that titin is phosphorylated in skinned myocardial tissues; this effect is exacerbated by pretreating with protein phosphatase 1. In vitro phosphorylation of recombinant protein representing titin's spring elements showed that PKCalpha targets the proline - glutamate - valine - lysine (PEVK) spring element. Furthermore, mass spectrometry in combination with site-directed mutagenesis identified 2 highly conserved sites in the PEVK region that are phosphorylated by PKCalpha (S11878 and S12022); when these 2 sites are mutated to alanine, phosphorylation is effectively abolished. Mechanical experiments with skinned left ventricular myocardium revealed that PKCalpha significantly increases titin-based passive tension, an effect that is reversed by protein phosphatase 1. Single molecule force-extension curves show that PKCalpha decreases the PEVK persistence length (from 1.20 nm to 0.55 nm), without altering the contour length, and using a serially-linked wormlike chain model we show that this increases titin-based passive force with a sarcomere length dependence that is similar to that measured in skinned myocardium after PKCalpha phosphorylation. CONCLUSIONS: PKC phosphorylation of titin is a novel and conserved pathway that links myocardial signaling and myocardial stiffness. PMID- 19679841 TI - Factors explaining excess stroke prevalence in the US Stroke Belt. AB - BACKGROUND AND PURPOSE: Higher risk and burden of stroke have been observed within the southeastern states (the Stroke Belt) compared with elsewhere in the United States. We examined reasons for these disparities using a large data set from a nationwide cross-sectional study. METHODS: Self-reported data from the 2005 and 2007 Behavioral Risk Factor Surveillance System were used (n=765,368). The potential contributors for self-reported stroke prevalence (n=27 962) were demographics (age, sex, geography, and race/ethnicity), socioeconomic status (education and income), common risk factors (smoking and obesity), and chronic diseases (hypertension, diabetes, and coronary heart disease). Multivariate logistic regression was used in the analysis. RESULTS: The age- and sex-adjusted OR comparing self-reported stroke prevalence in the 11-state Stroke Belt versus non-Stroke Belt region was 1.25 (95% CI, 1.19 to 1.31). Unequal black/white distribution by region accounted for 20% of the excess prevalence in the Stroke Belt (OR reduced to 1.20; 1.15 to 1.26). Approximately one third (32%) of the excess prevalence was accounted either by socioeconomic status alone or by risk factors and chronic disease alone (OR, 1.12). The OR was further reduced to 1.07 (1.02 to 1.13) in the fully adjusted logistic model, a 72% reduction. CONCLUSIONS: Differences in socioeconomic status, risk factors, and prevalence of common chronic diseases account for most of the regional differences in stroke prevalence. PMID- 19679840 TI - LIM kinase 1 promotes endothelial barrier disruption and neutrophil infiltration in mouse lungs. AB - RATIONALE: Disruption of endothelial barrier function and neutrophil-mediated injury are two major mechanisms underlying the pathophysiology of sepsis-induced acute lung injury (ALI). Recently we reported that endotoxin induced activation of RhoA in mice lungs that led to the disruption of endothelial barrier and lung edema formation; however, the molecular mechanism of this phenomenon remained unknown. OBJECTIVE: We reasoned that LIMK1, which participates in the regulation of endothelial cell contractility and is activated by RhoA/Rho kinase pathway, could mediate RhoA-dependent disruption of endothelial barrier function in mouse lungs during ALI. And if that is the case, then attenuation of endothelial cell contractility by downregulating LIMK1 may lead to the enhancement of endothelial barrier function, which could protect mice from endotoxin-induced ALI. METHODS AND RESULTS: Here we report that LIMK1 deficiency in mice significantly reduced mortality induced by endotoxin. Data showed that lung edema formation, lung microvascular permeability, and neutrophil infiltration into the lungs were suppressed in limk1(-/-) mice. CONCLUSIONS: We identified that improvement of endothelial barrier function along with impaired neutrophil chemotaxis were the underlying mechanisms that reduced severity of ALI in limk1(-/-) mice, pointing to a new therapeutic target for diseases associated with acute inflammation of the lungs. PMID- 19679842 TI - Association of symptoms of chronic bronchitis and frequent flu-like illnesses with stroke. AB - BACKGROUND AND PURPOSE: Acute and several chronic infectious diseases increase the risk of stroke. We tested the hypothesis that chronic bronchitis and frequent flu-like illnesses are independently associated with the risk of stroke or transient ischemic attack (TIA). METHODS: We assessed symptoms of chronic bronchitis, frequency of flu-like illnesses, and behavior during acute febrile infection in 370 consecutive patients with ischemic or hemorrhagic stroke or TIA and 370 age- and sex-matched control subjects randomly selected from the population. RESULTS: Cough with phlegm during > or = 3 months per year (grade 2 symptoms of chronic bronchitis) was associated with stroke or TIA independent from smoking history, other risk factors, and school education (odds ratio [OR] 2.63, 95% confidence interval [CI] 1.17 to 5.94; P=0.021). There was also an independent association between frequent flu-like infections (>2 per yr) and stroke/TIA (OR 3.54; 95% CI 1.52 to 8.27; P=0.003). Simultaneous assessment of chronic bronchitis and frequent flu-like infections did not attenuate the effect of either factor. Patients reported more often than control subjects to continue to work despite febrile infection (OR 3.68, 95% CI 1.80 to 7.52, multivariate analysis). CONCLUSIONS: Our results suggest that chronic bronchitis is among those chronic infections that increase the risk of stroke. Independent from chronic bronchitis, a high frequency of flu-like illnesses may also be a stroke risk factor. Infection-related behavior may differ between stroke patients and control subjects. PMID- 19679843 TI - Do endothelin-receptor antagonists prevent delayed neurological deficits and poor outcomes after aneurysmal subarachnoid hemorrhage?: a meta-analysis. AB - BACKGROUND AND PURPOSE: Delayed ischemic neurological deficits (DINDs) contribute to poor outcomes after aneurysmal subarachnoid hemorrhage (SAH). Endothelin-1 is an important mediator involved in the development of vasospasm. METHODS: We performed a systematic review and meta-analysis of randomized controlled trials assessing the use of endothelin-receptor antagonists (ETRAs) in patients with SAH. RESULTS: Three studies met eligibility criteria, enrolling 867 patients. ETRAs significantly reduced the occurrence of DINDs (OR 0.68 [0.49 to 0.95]) and radiographic vasospasm (OR 0.31 [0.19 to 0.49]), but did not have any impact on mortality (OR 1.09 [0.69 to 1.72]) or poor neurological outcomes (OR 0.87 [0.63 to 1.20]). Any benefit of ETRAs may have been partially offset by adverse effects, including hypotension(OR 2.39 [1.37 to 4.17]) and pulmonary complications (OR 2.12 [1.51 to 2.98]). CONCLUSIONS: Although ETRAs reduce radiographic vasospasm and DINDs, there is currently no evidence that they improve outcomes. PMID- 19679844 TI - Optimizing screening and management of asymptomatic coronary artery disease in patients with stroke and patients with transient ischemic attack. AB - BACKGROUND AND PURPOSE: The feasibility of implementing an expert consensus guideline recommending use of a stroke patient's profile to manage undiagnosed coronary artery disease remains unclear. METHODS: Following a guideline-based algorithm, we screened consecutive patients with ischemic stroke and patients with transient ischemic attack for asymptomatic coronary artery disease using the Framingham Heart Study Coronary Risk Score (FCRS) cutoff of high risk (> or = 20%) for experiencing a hard coronary artery disease event over a 10-year period. Patients with high FCRS received dobutamine stress echocardiogram outpatient screening, additional treatment (beta-blocker), or further management (cardiologist referral). RESULTS: From July 2004 to September 2007, among 693 patients, 501 (72%) met study criteria, of which 80 (16%) had FCRS > or = 20%. Elevated serum glucose, nonhigh-density lipoprotein, triglycerides, homocysteine, glycosylated hemoglobin as well as large vessel atherosclerotic stroke mechanism were more frequent in high versus low FCRS patients (P<0.05). Among high FCRS patients, 35 (44%) had dobutamine stress echocardiogram performed. Leading reasons for dobutamine stress echocardiogram nonperformance were patient noncompliance (42%) and primary care physician refusal (33%). CONCLUSIONS: Screening for coronary artery disease risk using FCRS is feasible in hospitalized patients with stroke, but outpatient adherence to stress testing is challenging largely due to patient and primary care physician-related factors. PMID- 19679845 TI - Geography, structure, and evolution of diffusion and perfusion lesions in Diffusion and perfusion imaging Evaluation For Understanding Stroke Evolution (DEFUSE). AB - BACKGROUND AND PURPOSE: The classical representation of acute ischemic lesions on MRI is a central diffusion-weighted imaging (DWI) lesion embedded in a perfusion weighted imaging (PWI) lesion. We investigated spatial relationships between final infarcts and early DWI/PWI lesions before and after intravenous thrombolysis in the Diffusion and perfusion imaging Evaluation For Understanding Stroke Evolution (DEFUSE) study. METHODS: Baseline and follow-up DWI and PWI lesions and 30-day fluid-attenuated inversion recovery scans of 32 patients were coregistered. Lesion geography was defined by the proportion of the DWI lesion superimposed by a Tmax (time when the residue function reaches its maximum) >4 seconds PWI lesion; Type 1: >50% overlap and Type 2: < or = 50% overlap. Three dimensional structure was dichotomized into a single lesion (one DWI and one PWI lesion) versus multiple lesions. Lesion reversal was defined by the percentage of the baseline DWI or PWI lesion not superimposed by the early follow-up DWI or PWI lesion. Final infarct prediction was estimated by the proportion of the final infarct superimposed on the union of the DWI and PWI lesions. RESULTS: Single lesion structure with Type 1 geography was present in only 9 patients (28%) at baseline and 4 (12%) on early follow-up. In these patients, PWI and DWI lesions were more likely to correspond with the final infarcts. DWI reversal was greater among patients with Type 2 geography at baseline. Patients with multiple lesions and Type 2 geography at early follow-up were more likely to have early reperfusion. CONCLUSION: Before thrombolytic therapy in the 3- to 6-hour time window, Type 2 geography is predominant and is associated with DWI reversal. After thrombolysis, both Type 2 geography and multiple lesion structure are associated with reperfusion. PMID- 19679846 TI - Reliability of the modified Rankin Scale: a systematic review. AB - BACKGROUND AND PURPOSE: A perceived weakness of the modified Rankin Scale is potential for interobserver variability. We undertook a systematic review of modified Rankin Scale reliability studies. METHODS: Two researchers independently reviewed the literature. Crossdisciplinary electronic databases were interrogated using the following key words: Stroke*; Cerebrovasc*; Modified Rankin*; Rankin Scale*; Oxford Handicap*; Observer variation*. Data were extracted according to prespecified criteria with decisions on inclusion by consensus. RESULTS: From 3461 titles, 10 studies (587 patients) were included. Reliability of modified Rankin Scale varied from weighted kappa=0.95 to kappa=0.25. Overall reliability of mRS was kappa=0.46; weighted kappa=0.90 (traditional modified Rankin Scale) and kappa=0.62; weighted kappa=0.87 (structured interview). CONCLUSION: There remains uncertainty regarding modified Rankin Scale reliability. Interobserver studies closest in design to large-scale clinical trials demonstrate potentially significant interobserver variability. PMID- 19679847 TI - Multi-ethnic genetic association study of carotid intima-media thickness using a targeted cardiovascular SNP microarray. AB - BACKGROUND AND PURPOSE: Identification of subclinical atherosclerosis by ultrasonographic measurement of carotid intima-media thickness (IMT) is a validated tool, in conjunction with traditional risk factors, for clinical assessment of cardiovascular disease risk. IMT has also been recognized as a quantitative measure of cardiovascular disease progression in asymptomatic individuals, and many candidate gene association studies have attempted to identify genetic variants associated with interindividual differences in IMT with limited success. We sought to test the association between subclinical atherosclerosis measured by IMT and approximately 50,000 SNPs, densely mapping approximately 2100 genes found on the gene-centric Illumina cardiovascular disease beadchip in a multi-ethnic population-based sample. METHODS: IMT was measured by B-mode ultrasound and DNA was collected from a population-based sample of South Asian (n=328), Chinese (n=302), and European Caucasian (n=268) participants. Genetic association was measured using multivariate linear regression including adjustment for covariates. RESULTS: The most robust association across all models tested was observed for a SNP (rs3791398) in histone deacetylase 4 (HDAC4; P=1.8e-5 to P=3.6e-5), while another strong association signal was observed with natriuretic peptide receptor a/guanylate cyclase A (NPR1) (rs10082235, P=5.4e-5). Seven of 13 previously reported functional candidate genes contained a SNP that was marginally associated (0.01 < P < or = 0.05). CONCLUSION: This initial multi-ethnic high-density association study of carotid IMT suggests some novel loci requiring further evaluation in follow-up studies. PMID- 19679848 TI - Intraventricular fibrinolysis and lumbar drainage for ventricular hemorrhage. AB - BACKGROUND AND PURPOSE: Both intraventricular fibrinolysis (IVF) and lumbar drainage (LD) may reduce the need for exchange of external ventricular drainage (EVD) and shunt surgery in patients with intracerebral hemorrhage and severe intraventricular hemorrhage. We investigated the feasibility and safety of IVF followed by early LD for the treatment of posthemorrhagic hydrocephalus. METHODS: This prospective study included patients with spontaneous ganglionic intracerebral hemorrhage and severe intraventricular hemorrhage with acute obstructive posthemorrhagic hydrocephalus who received an EVD (n=32). The treatment algorithm started with IVF (4 mg recombinant tissue plasminogen activator every 12 hours) until clearance of the third and fourth ventricles from blood. Thereupon, EVD was clamped and if clamping was unsuccessful, communicating posthemorrhagic hydrocephalus was assumed and LD placed. EVD was removed if there was neither an increase of intracranial pressure nor ventricle enlargement on CT. A ventriculoperitoneal shunt was indicated if "LD weaning" was unsuccessful for >10 days. Outcome was assessed at 90 and 180 days using the modified Rankin Scale. RESULTS: IVF resulted in fast clearance of the third and fourth ventricles (73+/-50 hours). However, early EVD removal was only possible in 4 patients. The remaining 28 patients developed communicating posthemorrhagic hydrocephalus. In all of these patients, early LD was capable to replace EVD. EVD exchange was not necessary and EVD duration was 105+/-59 hours. Only one patient required a ventriculoperitoneal shunt. At 180 days, 20 (62.5%) patients had a good (modified Rankin Scale 0 to 3) outcome and 5 (15.6%) patients had died. One patient had asymptomatic ventricular rebleeding. CONCLUSIONS: In patients with secondary intraventricular hemorrhage and posthemorrhagic hydrocephalus, the combined treatment approach of IVF and early LD is safe and feasible, avoids EVD exchange, and may markedly reduce the need for shunt surgery. PMID- 19679849 TI - Defeating normal thermoregulatory defenses: induction of therapeutic hypothermia. AB - Therapeutic hypothermia may be useful in various circumstances including stroke. However, core body temperature is normally tightly regulated. Even mild hypothermia in conscious subjects thus provokes vigorous thermoregulatory defenses which are potentially harmful in fragile patients. Furthermore, thermoregulatory responses are effective, which reduces the rate at which hypothermia can be induced. Drugs are thus often given to blunt normal thermoregulatory defenses. General anesthetics profoundly impair thermoregulatory control, but prolonged general anesthesia is rarely practical or appropriate. A variety of other drugs have therefore been evaluated. Most opioids only slightly impair thermoregulatory defenses, but meperidine is considerably more effective than equipotent doses of other opioids. The central alpha-2 agonists clonidine and dexmedetomidine are also useful. However, the best overall approach to inducing thermal tolerance appears to be a combination of buspirone and meperidine, which reduces the core temperature triggering shivering to about 33.5 degrees C in doses that maintain adequate ventilation. PMID- 19679850 TI - Expanded spectrum of gene causing both hypertrophic cardiomyopathy and dilated cardiomyopathy. PMID- 19679851 TI - Bottom-line reality tempers cancer researchers' excitement. PMID- 19679852 TI - StatBite: Stimulus funding: where does the NCI propose to invest? PMID- 19679853 TI - Classification as diagnostic reasoning. AB - An ongoing goal in the field of categorization has been to determine how objects' features provide evidence of membership in one category versus another. Well known findings include that feature diagnosticity is a function of how often the feature appears in category members versus nonmembers, their perceptual salience, how features are used in support of inferences, and how observable features are related to other observable features. We tested how diagnosticity is affected by causal relations between observable and unobserved features. Consistent with our view of classification as diagnostic reasoning, we found that observable features are more diagnostic to the extent that they are caused by underlying features that define category membership, because the presence of the latter can be (causally) inferred from the former. Implications of these results for current views of conceptual structure and models of categorization are discussed. PMID- 19679854 TI - Induction with uncertain categories: When do people consider the category alternatives? AB - These three experiments examined how people make property inferences about exemplars whose category membership is uncertain. Participants were shown two categories and a novel exemplar with a feature that indicated that the exemplar was more likely to belong to one category (target) than to the other (nontarget). Participants then made categorization decisions and property inferences about the novel exemplar. In some conditions, property inferences could be made only by considering both target and nontarget categories. In other conditions, predictions could be based on both categories or on the target category alone. Consistent with previous studies (e.g., Murphy & Ross, 1994, 2005), we found that many people made predictions based only on consideration of the target category. However, the prevalence of such single-category reasoning was greatly reduced by highlighting the costs of neglecting nontarget alternatives and by asking for inferences before categorization decisions. The results suggest that previous work may have exaggerated the prevalence of single-category reasoning and that people may be more flexible in their use of multiple categories in property inference than has been previously recognized. PMID- 19679855 TI - The conceptual centrality of causal cycles. AB - How do causal cycles affect judgments of conceptual centrality? Generally, a feature is central to a concept to the extent that other features in the concept depend on it, thereby rendering it immutable from the concept (Sloman, Love, & Ahn, 1998). Previous research on conceptual centrality has focused primarily on features involved in four major types of dependency structures: simple cause effect relations, causal chains, common-cause structures, and common-effect structures. Causal cycles are a fifth type of dependency structure commonly reported in people's real-life concepts, yet to date, they have been relatively ignored in research on conceptual centrality. The results of six experiments suggest that previously held assumptions about the conceptual representation of cycles are incorrect. We discuss the implications of these findings for our understanding of theory-based concepts. PMID- 19679856 TI - Phonological and visual distinctiveness effects in syllogistic reasoning: implications for mental models theory. AB - Two experiments are reported in which the representational distinctiveness of terms within categorical syllogisms was manipulated in order to examine the assumption of mental models theory that abstract, spatially based representations underpin deduction. In Experiment 1, participants evaluated conclusion validity for syllogisms containing either phonologically distinctive terms (e.g., harks, paps, and fids) or phonologically nondistinctive terms (e.g., fuds, fods, and feds). Logical performance was enhanced with the distinctive contents, suggesting that the phonological properties of syllogism terms can play an important role in deduction. In Experiment 2, participants received either the phonological materials from Experiment 1 or syllogisms involving distinctive or nondistinctive visual contents. Logical inference was again enhanced for the distinctive contents, whether phonological or visual in nature. Our findings suggest a broad involvement of multimodal information in syllogistic reasoning and question the assumed primacy of abstract, spatially organized representations in deduction, as is claimed by mental models theorists. PMID- 19679857 TI - Age differences in tracking characters during narrative comprehension. AB - Understanding a narrative situation depends on keeping track of multiple characters that enter and exit dynamically as the plot unfolds. We investigated age differences in this process during narrative comprehension. In Experiment 1, we used a probe recognition paradigm to examine the effect of age on the accessibility of a previous character when another character was subsequently introduced. In Experiment 2, reading time was measured to examine age differences in the encoding of a new character after another had already been introduced. Our findings show that older readers have particular difficulty both in accessing the initial character after a new character is introduced and in thoroughly encoding a new character while other characters inhabit the discourse world. We attribute these differences to age differences in working memory that make it difficult to access a backgrounded character when a new character is in focus and to distinctively encode a new character when maintenance of another character is already consuming attentional resources. PMID- 19679858 TI - Motivated comprehension regulation: vigilant versus eager metacognitive control. AB - The more accurately people assess their comprehension, the more likely they are to engage in study behaviors that precisely target gaps in their learning. However, comprehension regulation involves more than knowing when to implement a new study strategy; it also involves deciding which strategy will most effectively resolve one's confusion. In two experiments, we explored how people's motivational orientations influence which study strategies they select to regulate their comprehension. In Experiment 1, people who were motivated to vigilantly protect against potential mistakes (i.e., prevention-focused individuals) were more likely to adopt a rereading strategy than people who were motivated to eagerly pursue new learning opportunities (i.e., promotion-focused individuals). In Experiment 2, this difference in strategy use emerged specifically in response to confusing sentences that had been inserted into the text. Furthermore, by using rereading strategies to resolve their confusion, prevention-focused individuals performed better than promotion-focused individuals on a comprehension test and a transfer task. PMID- 19679859 TI - Spatial directions and situation model organization. AB - Do spatial directions, such as "to the right," influence the integration and segregation of information into situation models? According to a single-framework hypothesis, spatial location serves as an event framework, and spatial directions serve as relational information within that framework but do not establish separate sublocation frameworks. Alternatively, according to a fragmented framework hypothesis, spatial directions lead the larger framework to be broken down such that each direction is treated as a separate sublocation, thereby producing retrieval interference. In three experiments, people memorized sentences about objects in locations. The results support the fragmented framework hypothesis. Control conditions ruled out explanations based on the ease of memorization, retrieval demands, or sentence complexity. PMID- 19679860 TI - Global subjective memorability and the strength-based mirror effect in recognition memory. AB - Between-list manipulations of memory strength through repetition commonly generate a mirror effect, with more hits and fewer false alarms for strengthened items. However, this pattern is rarely seen with within-list manipulations of strength. In three experiments, we investigated the conditions under which a within-list mirror effect of strength (items presented once or thrice) is observed. In Experiments 1 and 2, we indirectly manipulated the overall subjective memorability of the studied lists by varying the proportion of nonwords. A within-list mirror effect was observed only in Experiment 2, in which a higher proportion of nonwords was presented in the study list. In Experiment 3, the presentation duration for each item (0.5 vs. 3 sec) was manipulated between groups with the purpose of affecting subjective memorability. A within-list mirror effect was observed only for the short presentation durations. Thus, across three experiments, we found the within-list mirror effect only under conditions of poor overall subjective memorability. We propose that when the overall subjective memorability is low, people switch their response strategy on an item-by-item basis and that this generates the observed mirror effect. PMID- 19679861 TI - Retrieval-induced forgetting and mental imagery. AB - In the present article, we present four experiments in which we examined whether mental imagery can initiate retrieval-induced forgetting. Participants were presented with word pairs (Experiments 1, 2, and 3) or narratives (Experiment 4) and then engaged in selective mental imagery about half of the details from half of the categories. The results indicated that mental imagery can produce the same pattern of impairment as retrieval practice (Experiment 1) and postevent questioning (Experiment 4). Additionally, mental imagery-invoked, retrieval induced forgetting was found for category cued recall (Experiments 1, 3, and 4) and cued recall (Experiment 2); it was found to dissipate across a 24-h delay, but only when there was no pre-delay test (Experiment 3). Such retrieval-induced forgetting was also found for imagining from the first-person and third-person perspectives (Experiment 4). From these findings, we suggest that the underlying retrieval processes behind mental imagery can initiate retrieval-induced forgetting. The findings are discussed in terms of inhibitory processes. PMID- 19679862 TI - How verbal memory loads consume attention. AB - According to a traditional assumption about working memory, participants retain a series of verbal items for immediate recall using covert verbal rehearsal, without much need for attention. We reassessed this assumption by imposing a speeded, nonverbal choice reaction time (CRT) task following the presentation of each digit in a list to be recalled. When the memory load surpassed a few items, performance on the speeded CRT task became increasingly impaired. This CRT task impairment depended only on attention-related components of working memory; it was not alleviated by the presence of an auditory memory trace that automatically helped the recall of items at the ends of spoken lists. We suggest that attention demanding refreshing of verbal stimuli occurs along with any covert rehearsal. PMID- 19679863 TI - Variation in working memory capacity, fluid intelligence, and episodic recall: a latent variable examination of differences in the dynamics of free recall. AB - A latent variable analysis was conducted to examine the nature of individual differences in the dynamics of free recall and cognitive abilities. Participants performed multiple measures of free recall, working memory capacity (WMC), and fluid intelligence (gF). For each free recall task, recall accuracy, recall latency, and number of intrusion errors were determined, and latent factors were derived for each. It was found that recall accuracy was negatively related to both recall latency and number of intrusions, and recall latency and number of intrusions were positively related. Furthermore, latent WMC and gF factors were positively related to recall accuracy, but negatively related to recall latency and number of intrusions. Finally, a cluster analysis revealed that subgroups of participants with deficits in focusing the search had deficits in recovering degraded representations or deficits in monitoring the products of retrieval. The results are consistent with the idea that variation in the dynamics of free recall, WMC, and gF are primarily due to differences in search set size, but differences in recovery and monitoring are also important. PMID- 19679864 TI - The interaction of word frequency and concreteness in immediate serial recall. AB - Word frequency and word concreteness are language attributes that have been shown to independently influence the recall of items in verbal short-term memory (STM). It has been argued that such effects are evidence for the action of long-term memory knowledge on STM traces. However, research to date has not investigated whether these variables interact in serial recall. In two experiments, we examined the behavior of these variables under factorial manipulation and demonstrated that the effect of word frequency is dependent on the level of concreteness of items. Serial recall performance is examined with reference to two explanatory approaches: Walker and Hulme's (1999) dual-redintegration account and language-based models of STM. The data indicate that language-based models are more compatible with the observed effects and challenge the view that frequency and concreteness effects in STM are the products of distinct mechanisms. PMID- 19679865 TI - Taboo words: the effect of emotion on memory for peripheral information. AB - In three experiments, we examined memory for peripheral information that occurred in the same context as emotion-inducing information. In the first two experiments, participants studied either a sentence (Experiment 1) or a pair of words (Experiments 2A-2C) containing a neutral peripheral word, as well as a neutral, negative-valence, or taboo word, to induce an emotional response. At retrieval, the participants were asked to recall the neutral peripheral word from a sentence fragment or emotion-inducing word cue. In Experiment 3, we presented word pairs at encoding and tested memory with associative recognition. In all three experiments, memory for peripheral words was enhanced when it was encoded in the presence of emotionally arousing taboo words but not when it was encoded in the presence of words that were only negative in valence. These data are consistent with priority-binding theory (MacKay et al., 2004) and inconsistent with the attention-narrowing hypothesis (Easterbrook, 1959), as well as with object-based binding theory (Mather, 2007). PMID- 19679866 TI - Enhancement and suppression effects resulting from information structuring in sentences. AB - Information structuring through the use of cleft sentences increases the processing efficiency of references to elements within the scope of focus. Furthermore, there is evidence that putting certain types of emphasis on individual words not only enhances their subsequent processing, but also protects these words from becoming suppressed in the wake of subsequent information, suggesting mechanisms of enhancement and suppression. In Experiment 1, we showed that clefted constructions facilitate the integration of subsequent sentences that make reference to elements within the scope of focus, and that they decrease the efficiency with reference to elements outside of the scope of focus. In Experiment 2, using an auditory text-change-detection paradigm, we showed that focus has similar effects on the strength of memory representations. These results add to the evidence for enhancement and suppression as mechanisms of sentence processing and clarify that the effects occur within sentences having a marked focus structure. PMID- 19679867 TI - Does visual speech information affect word segmentation? AB - We present an experiment in which we explored the extent to which visual speech information affects learners' ability to segment words from a fluent speech stream. Learners were presented with a set of sentences consisting of novel words, in which the only cues to the location of word boundaries were the transitional probabilities between syllables. They were exposed to this language through the auditory modality only, through the visual modality only (where the learners saw the speaker producing the sentences but did not hear anything), or through both the auditory and visual modalities. The learners were successful at segmenting words from the speech stream under all three training conditions. These data suggest that visual speech information has a positive effect on word segmentation performance, at least under some circumstances. PMID- 19679868 TI - Semantic transparency and masked morphological priming: the case of prefixed words. AB - In four lexical decision experiments, we investigated masked morphological priming with Dutch prefixed words. Reliable effects of morphological relatedness were obtained with visual primes and visual targets in the absence of effects due to pure form overlap. In certain conditions, priming effects were significantly greater with semantically transparent prefixed primes (e.g., rename-name) relative to the priming effects obtained with semantically opaque prefixed words (e.g., relate-late), even with very brief (40-msec) prime durations. With visual primes and auditory targets (cross-modal priming), significant facilitation was found in all related prime conditions, independent of whether or not primes were morphologically related to targets. The results are interpreted within a bimodal hierarchical model of word recognition in which morphological effects arise through the interplay of sublexical (morpho-orthographic) and supralexical (morpho-semantic) representations. The word stimuli from this study may be downloaded as supplemental materials from http://mc.psychonomic journals.org/content/supplemental. PMID- 19679869 TI - Updating objects in visual short-term memory is feature selective. AB - The purpose of this study was to examine whether the process of updating information in visual short-term memory (VSTM) is object based. We investigated whether modifying the memory of one feature of an object would automatically promote refreshing the memory of all of its other features. The results showed that the facilitative effect of updating was specific to the updated feature of an object and did not spread to its nonupdated features. This feature-selective effect suggests that updating VSTM is not object based (Experiment 1), even though storage was object based (Experiment 2). Control experiments ruled out strategy-based (Experiment 3) and stimulus-related (Experiments 4-6) accounts. Feature-selective updating may indicate that the mechanism used to modify the contents of memory may have a different basis than that used to encode or store information in memory. PMID- 19679870 TI - Are spatial and dimensional attention separate? evidence from Posner, Stroop, and Eriksen tasks. AB - Do various operational definitions of visual attention tap the same underlying process? To address this question, we probed visual selective attention using orientation of attention, flanker, and Stroop tasks. These were embedded in combined designs that enabled assessment of each effect, as well as their interaction. For the orientation task, performance was poorer at unexpected than at expected locations. The flanker effects also differed across the two locations. In contrast, the Stroop effects were comparable at expected and unexpected locations. We conclude that spatial attention (tapped by the orientation and the flanker tasks) and dimensional attention (tapped by the Stroop task) engage separate processes of visual selection, both of which are needed in normal attention processing. PMID- 19679871 TI - Impaired color word processing at an unattended location: evidence from a Stroop task combined with inhibition of return. AB - A Stroop task with separate color bar and color word stimuli was combined with an inhibition-of-return procedure to examine whether visual attention modulates color word processing. In Experiment 1, the color bar was presented at the cued location and the color word at the uncued location, or vice versa, with a 100- or 1,050-msec stimulus onset asynchrony (SOA) between cue and Stroop stimuli. In Experiment 2, on Stroop trials, the color bar was presented at a central fixated location and the color word at a cued or uncued location above or below the color bar. In both experiments, with a 100-msec SOA, the Stroop effect was numerically larger when the color word was displayed at the cued location than when it was displayed at the uncued location, but with the 1,050-msec SOA, this relation between Stroop effect magnitude and location was reversed. These results provide evidence that processing of the color word in the Stroop task is modulated by the location to which visual attention is directed. PMID- 19679872 TI - LookSeq: a browser-based viewer for deep sequencing data. AB - Sequencing a genome to great depth can be highly informative about heterogeneity within an individual or a population. Here we address the problem of how to visualize the multiple layers of information contained in deep sequencing data. We propose an interactive AJAX-based web viewer for browsing large data sets of aligned sequence reads. By enabling seamless browsing and fast zooming, the LookSeq program assists the user to assimilate information at different levels of resolution, from an overview of a genomic region to fine details such as heterogeneity within the sample. A specific problem, particularly if the sample is heterogeneous, is how to depict information about structural variation. LookSeq provides a simple graphical representation of paired sequence reads that is more revealing about potential insertions and deletions than are conventional methods. PMID- 19679873 TI - Essential roles for early growth response transcription factor Egr-1 in tissue fibrosis and wound healing. AB - The early growth response gene (Egr-1) codes for a zinc finger transcription factor that has important roles in the regulation of cell growth, differentiation, and survival. Aberrant Egr-1 expression is implicated in carcinogenesis, inflammation, atherosclerosis, and ischemic injury. We reported previously that normal fibroblasts stimulated by transforming growth factor-ss showed rapid and transient induction of Egr-1. Moreover, we observed that tissue expression of Egr-1 was elevated in patients with scleroderma, which suggests that Egr-1 may be involved in tissue repair and fibrosis. Here, we investigated matrix remodeling and wound healing in mice harboring gain of function or loss of function mutations of Egr-1. Using the model of bleomycin-induced scleroderma, we found that the early influx of inflammatory cells into the skin and lungs, and the subsequent development of fibrosis in these organs, were markedly attenuated in Egr-1 null mice. Furthermore, full-thickness incisional skin wound healing was impaired, and skin fibroblasts lacking Egr-1 showed reduced migration and myofibroblast transdifferentiation in vitro. In contrast, transgenic mice with fibroblast-specific Egr-1 overexpression showed exuberant tissue repair, with enhanced collagen accumulation and increased tensile strength of incisional wounds. Together, these results point to the fundamental role that Egr-1 plays in the regulation of transforming growth factor-ss-dependent physiological and pathological matrix remodeling. PMID- 19679875 TI - Fenfluramine disrupts the mitral valve interstitial cell response to serotonin. AB - Serotonin (5HT) receptor signaling and 5HT-related agents, such as the anorexogen fenfluramine (Fen), have been associated with heart valve disease. We investigated the hypothesis that Fen may disrupt mitral valve interstitial cell (MVIC) homeostasis through its effects on mitogenesis and extracellular matrix biosynthesis. Normal and myxomatous mitral valves, both human and canine, were harvested, and primary MVIC cultures were established. 5HT caused increased phosphorylation of extracellular signal-related kinase in MVIC; Fen alone did not. However, Fen combined with 5HT increased the level of MVIC extracellular signal-related kinase, when compared with 5HT alone. In addition, MVIC mitogenesis per (3)H-thymidine ((3)HTdR) demonstrated a 5HT dose-dependent increase, with no effect of Fen alone. In contrast, Fen combined with 5HT inhibited the MVIC (3)HTdR response when compared with 5HT alone. Furthermore, fluoxetine, a 5HT transporter inhibitor, while having no effect alone, suppressed Fen-5HT (3)HTdR inhibition when administered with Fen plus 5HT. Finally, MVIC incorporations of (3)H-proline and (3)H-glucosamine, measures of extracellular matrix collagen and glycosaminoglycan respectively, were increased with 5HT alone; however, Fen did not affect MVIC glycosaminoglycan or collagen either alone or in combination with 5HT. Taken together, the ratios of (3)H-proline or (3)H-glycosaminoglycan to (3)HTdR in MVIC, normalized to 5HT alone, demonstrated a significant imbalance of extracellular matrix production versus proliferation in MVIC cultures with Fen plus 5HT exposure. This imbalance may explain in part the pathophysiology of Fen-related mitral valve disease. PMID- 19679874 TI - Characterization of RAGE, HMGB1, and S100beta in inflammation-induced preterm birth and fetal tissue injury. AB - Immune activation represents an adaptive reaction triggered by both noxious exogenous (microbes) and endogenous [high mobility group box-1 protein (HMGB1), S100 calcium binding proteins] inducers of inflammation. Cell stress or necrosis lead the release of HMGB1 and S100 proteins in the extracellular compartment where they act as damage-associated molecular pattern molecules (or alarmins) by engaging the receptor for advanced glycation end-products (RAGE). Although the biology of RAGE is dictated by the accumulation of damage-associated molecular pattern molecules at sites of tissue injury, the role of RAGE in mediating antenatal fetal injury remains unknown. First, we studied the relationships at birth between the intensity of human fetal inflammation and sRAGE (an endogenous RAGE antagonist), HMGB1, and S100beta protein. We found significantly lower sRAGE in human fetuses that mounted robust inflammatory responses. HMGB1 levels correlated significantly with levels of interleukin-6 and S100beta in fetal circulation. We then evaluated the levels and areas of tissue expression of RAGE, HMGB1, and S100beta in specific organs of mouse fetuses on E16. Using an animal model of endotoxin-induced fetal damage and preterm birth, we determined that inflammation induces a significant change in expression of RAGE and HMGB1, but not S100beta, at sites of tissue damage. Our findings indicate that RAGE and HMGB1 may be important mediators of cellular injury in fetuses delivered in the setting of inflammation-induced preterm birth. PMID- 19679876 TI - A novel organotypic model mimics the tumor microenvironment. AB - Carcinoma cell invasion is traditionally studied in three-dimensional organotypic models composed of type I collagen and fibroblasts. However, carcinoma cell behavior is affected by the various cell types and the extracellular matrix (ECM) in the tumor microenvironment. In this study, a novel organotypic model based on human uterine leiomyoma tissue was established and characterized to create a more authentic environment for carcinoma cells. Human tongue squamous cell carcinoma cells (HSC-3) were cultured on top of either collagen or myoma. Organotypic sections were examined by immunohistochemistry and in situ hybridization. The maximal invasion depth of HSC-3 cells was markedly increased in myomas compared with collagen. In myomas, various cell types and ECM components were present, and the HSC-3 cells only expressed ECM molecules in the myoma model. Organotypic media were analyzed by radioimmunoassay, zymography, or Western blotting. During carcinoma cell invasion, matrix metalloprotease-9 production and collagen degradation were enhanced particularly in the myoma model. To evaluate the general applicability of the myoma model, several oral carcinoma, breast carcinoma, and melanoma cell lines were cultured on myomas and found to invade in highly distinct patterns. We conclude that myoma tissue mimics the native tumor microenvironment better than previous organotypic models and possibly enhances epithelial-to-mesenchymal transition. Thus, the myoma model provides a promising tool for analyzing the behavior of carcinoma cells. PMID- 19679877 TI - Reduced phosphoinositide 3-kinase (p110alpha) activation increases the susceptibility to atrial fibrillation. AB - Atrial fibrillation (AF) is the most common sustained arrhythmia presenting at cardiology departments. A limited understanding of the molecular mechanisms responsible for the development of AF has hindered treatment strategies. The purpose of this study was to assess whether reduced activation of phosphoinositide 3-kinase (PI3K, p110alpha) makes the compromised heart susceptible to AF. Risk factors for AF, including aging, obesity, and diabetes, have been associated with insulin resistance that leads to depressed/defective PI3K signaling. However, to date, there has been no link between PI3K(p110alpha) and AF. To address this question, we crossed a cardiac-specific transgenic mouse model of dilated cardiomyopathy (DCM) with a cardiac-specific transgenic mouse expressing a dominant negative mutant of PI3K (dnPI3K; reduces PI3K activity). Adult ( approximately 4.5 months) double-transgenic (dnPI3K-DCM), single transgenic (DCM-Tg, dnPI3K-Tg), and nontransgenic mice were subjected to morphological, functional/ECG, microarray, and biochemical analyses. dnPI3K-DCM mice developed AF and had depressed cardiac function as well as greater atrial enlargement and fibrosis than DCM-Tg mice. AF was not detected in other groups. Aged DCM-Tg mice ( approximately 15 months) with a similar phenotype to dnPI3K DCM mice (4.5 months) did not develop AF, suggesting loss of PI3K activity directly contributed to the AF phenotype. Furthermore, increasing PI3K activity reduced atrial fibrosis and improved cardiac conduction in DCM-Tg mice. Finally, in atrial appendages from patients with AF, PI3K activation was lower compared with tissue from patients in sinus rhythm. These results suggest a link between PI3K(p110alpha) and AF. PMID- 19679878 TI - Beta-catenin activation promotes liver regeneration after acetaminophen-induced injury. AB - Acute liver failure (ALF) remains a disease with poor patient outcome. Improved prognosis is associated with spontaneous liver regeneration, which supports the relevance of exploring 'regenerative' therapies. Therefore, the role of the Wnt/beta-catenin pathway in liver regeneration following ALF was investigated. ALF was induced in mice by acetaminophen overdose, which is also a leading cause of liver failure in patients. beta-catenin distribution was also studied in liver sections from acetaminophen-induced ALF patients. A nonlethal dose of acetaminophen, which induces liver regeneration, led to stabilization and activation of beta-catenin for 1 to 12 hours. These data were also verified by increased expression of the beta-catenin surrogate target glutamine synthetase. Beta-catenin activation occurred secondary to the inactivation of glycogen synthase kinase-3beta and an increase in levels of casein kinase 2alpha, and led to increased cyclin-D1, another known beta-catenin target. These observations were next substantiated in beta-catenin conditional-null mice (beta-catenin null), which show dampened regeneration after acetaminophen injury following induction of CYP2e1/1a2 expression. In light of decreased acetaminophen injury in beta-catenin-null mice despite CYP induction, equitoxic studies in control mice were performed. Significant differences in regeneration persisted following comparable injury in beta-catenin-null and control animals. Retrospective analysis of liver samples from acetaminophen-overdose patients demonstrated a positive correlation between nuclear beta-catenin, proliferation, and spontaneous liver regeneration. Thus, our studies demonstrate early activation of beta catenin signaling during acetaminophen-induced injury, which contributes to hepatic regeneration. PMID- 19679879 TI - Hypercholesterolemic mice exhibit lymphatic vessel dysfunction and degeneration. AB - Lymphatic vessels are essential for lipid absorption and transport. Despite increasing numbers of observations linking lymphatic vessels and lipids, little research has been devoted to address how dysregulation of lipid balance in the blood, ie, dyslipidemia, may affect the functional biology of lymphatic vessels. Here, we show that hypercholesterolemia occurring in apolipoprotein E-deficient (apoE(-/-)) mice is associated with tissue swelling, lymphatic leakiness, and decreased lymphatic transport of fluid and dendritic cells from tissue. Lymphatic dysfunction results in part from profound structural abnormalities in the lymphatic vasculature: namely, initial lymphatic vessels were greatly enlarged, and collecting vessels developed notably decreased smooth muscle cell coverage and changes in the distribution of lymphatic vessel endothelial hyaluronic acid receptor-1 (LYVE-1). Our results provide evidence that hypercholesterolemia in adult apoE(-/-) mice is associated with a degeneration of lymphatic vessels that leads to decreased lymphatic drainage and provides an explanation for why dendritic cell migration and, thus, immune priming, are compromised in hypercholesterolemic mice. PMID- 19679880 TI - Neuroendocrine cancer-specific up-regulating mechanism of insulin-like growth factor binding protein-2 in small cell lung cancer. AB - Small cell lung cancer (SCLC) exhibits insulin-like growth factor-dependent growth. SCLC is the most aggressive among known in vivo lung cancers, whereas in vitro growth of SCLC is paradoxically slow as compared with that of non-SCLC (NSCLC). In this study, we demonstrate that SCLC cells overexpress insulin-like growth factor binding protein (IGFBP)-2 via NeuroD, a neuroendocrine cell specific transcription factor. Chromatin immunoprecipitation, electrophoretic mobility shift, and IGFBP-2 promoter assays all revealed that NeuroD binds to the E-box in the 5'-untranslated region of IGFBP-2. A NeuroD transgene in both airway epithelial and NSCLC cells up-regulated the transcription of IGFBP-2 and retarded cell growth. Recombinant IGFBP-2 repressed the growth of both airway epithelial and NSCLC cells in a dose-dependent manner. A NeuroD-specific small interfering RNA repressed IGFBP-2 expression in SCLC, and neutralization of IGFBP-2 and an IGFBP-2-specific small interfering RNA increased SCLC cell growth. Pathological samples of SCLC also expressed IGFBP-2 abundantly, as compared with NSCLC, and showed only rare (8%) IGFBP-2 promoter methylation, whereas the IGFBP-2 promoter was methylated in 71% of adenocarcinomas and 29% of squamous cell carcinomas. These findings suggest that 1) SCLC has an IGFBP-2 overexpression mechanism distinct from NSCLC, 2) secreted IGFBP-2 contributes to the slow growth of SCLC in vitro, and 3) the epigenetic alterations in the IGFBP-2 promoter contribute to the striking differences in IGFBP-2 expression between SCLC and NSCLC in vivo. PMID- 19679881 TI - The targeting of the sole cyclin D1 is not adequate for mantle cell lymphoma and myeloma therapies. PMID- 19679882 TI - Pediatric follicular lymphoma--a clinico-pathological study of a population-based series of patients treated within the Non-Hodgkin's Lymphoma--Berlin-Frankfurt Munster (NHL-BFM) multicenter trials. AB - Background Pediatric follicular lymphoma has recently been recognized as a novel variant of follicular lymphoma in the World Health Organization classification of lymphomas. Given the rarity of the disease, histopathological and genetic data on this type of lymphoma are still scarce. DESIGN AND METHODS: We analyzed 25 cases of pediatric follicular lymphoma (patients aged 0.05). Twin types: Of the twins with CHD 34% were male-female pairs, 26% were male-male pairs and 40% were female-female pairs. Age at diagnosis: Thirteen patients were diagnosed in the first week of life and 20 from one week to five months of age. After six months of age only two patients have been diag notnosed with CHD. TREATMENT: Twelve patients have been operated for CHD, one patient was treated in an interventional cardiac catheterization and 10 patients received medical treatment. Mortality: Six twins died as a consequence of their heart defect (17%). The mortality rate was significantly higher among twins with major CHD compared to the control population (p<0.005). CONCLUSION: Congenital heart defects are more common among twins compared to the control group. They were also more common in the latter half of the investigation period. The mortality rate is also higher among the twins. In 1991 in titero fertilization was started in Iceland resulting in increased number of twins and at the same time increase in incidence of CHD is seen. Further studies are required to clarify this. PMID- 19679919 TI - [Tuberculous infection and tuberculosis in the foreign born in Iceland.]. AB - OBJECTIVE: The purpose of the study was to find the incidence and prevalence of tuberculosis among the foreign-born in Iceland. MATERIAL AND METHODS: The study material was obtained from (a) the National TB register and (b) the files of the Immigration Office on recidency applicants in 1995. RESULTS: In 1975-1996 there were 32 cases of TB in the foreign-born out of a total of 468. The proportion of cases among the foreign-born rose significantly during the period (p<0.001). In 22 years the incidence of TB among the foreign-born was 18.0 but 8.4 among those born in Iceland (p<0.001). The incidence of TB in Asian-born was 173.7, or 21 times that among those born in Iceland (p<0.001). The second highest incidence (18.8) was in those born in North and South America. Tuberculosis usually appeared within five years of immigration. During 1995 the 559 applicants for recidence permit provided health certificates. Of these 363 had a tuberculin skin test (TST) and 42% were positive. The corresponding figure for those born in Africa was 68%, in Asia 58% and in East Europa 50%. Most of those with positive TST had a chest x-ray but also 23% of the others. 26.2% had neither a TST nor a chest x-ray but still received a health certificate. Only 33% of those positive received isoniazide to eliminate infection and 88% of these completed at least six months of isoniazide treatment. CONCLUSION: A TST is an indispensable part of health screening for immigrants and also a chest x-ray when appropriate. Treatment of TB infection should be used more often. PMID- 19679920 TI - [Use and abuse of alcohol among elderly women in Iceland. Comparison of clinical symptoms of abuse with liver function tests and mean corpuscular volume.]. AB - OBJECTIVE: This study was conducted to investigate the prevalence of alcohol abuse and dependency among elderly women in Iceland and to make an assessment of the sensitivity and specificity of liver function tests and mean corpuscular volume (MCV) in the diagnosis of alcohol abuse. METHODS: The group was made of 280 women (age 55-74 years, mean age 61.9 years) from the Reykjavik Study. The diagnoses were based on a questionnaire (eight symptoms) which has been used in epidemiological studies of alcoholism in Iceland since 1974. To this questionnaire there were added five questions aiding to the diagnosis of the lifetime prevalence rate of abuse. The CAGE questionnaire is implicated in this scale of 13 symptoms. RESULTS: Among 263 respondents (93.9%), there were 156 consumers (59.3%). Of the consumers 16% had at least one symptom indicating alcohol abuse. The diagnosis of abuse was made when three symptoms of eight occurred together. Abuse occured among 2.6% of consumers (1.5% of respondents). Symptom triad of dependency occurred among 1.3% of consumers (0.8% of respondents). Heavy drinking was equally frequent as dependency. Mean values of gamma glutamyl transferase (G-GT) and MCV increase with increasing number of symptoms of abuse. The sensitivity of G-GT and MCV is low (20%) but the specificity is high (94%) for both. Bilirubin and alkaline phosphatase do neither have a sensitivity nor specificity for the diagnosis of abuse. CONCLUSION: These findings indicate that these blood tests are not a reliable method for the screening of alcohol abuse among elderly women. A questionnaire searching for clinical symptoms of abuse is the best method for the diagnosis of abuse. Among elderly women, the symptom of guilty feelings connected to alcohol consumption, is the most sensitive indicator of abuse. But there are probably more women that experience guilty feelings than those which can be considered alcohol abusers. PMID- 19679921 TI - [Subcutaneous metastasis after laparoscopic cholecystectomy in a patient with unsuspected adenocarcinoma of the pancreas.]. AB - A 63 year old patient underwent uneventful laparoscopic cholecystectomy in 1994. The patient had a long history of biliary colic after fatty meals. The chief presenting symptom was pain localized in the epigastrium radiating to the back and later distributing to the whole abdomen. The patient also had a history of constipation, but no other symptoms were noted. An ultrasonogram of the liver, gall bladder and pancreas was reported to show calculi in the gall bladder but otherwise normal findings. The laparascopic cholecystectomy was uneventful with discharge the following day. The symptoms however did not disappear, changing in character, locating at the center of the abdomen. The patient began to lose appetite with bouts of diarrhea. The symptoms gradually increased and the patient was admitted to the hospital. Upon arrival the patient was found to have diffuse abdominal pain with a painful swelling of the umbilical trocar site. Incarcerated hernia was suspected, but proved to be a mass at exploration. Pathologic examination disclosed a metastatic adenocarcinoma. A similar but smaller mass was also discovered in the epigastric trocar site. CT scan showed a pancreatic carcinoma of the corpus with infiltration. The patient deteriorated rapidly and died four months after the diagnosis of pancreatic cancer. PMID- 19679922 TI - [Cutaneous leishmaniasis. A case report.]. AB - Cutaneous leishmaniasis is a common infection in South America and the Middle East. A 20 year Icelander was infected with leishmaniasis while travelling in South America. Treatment with the antimonial sodium stibogluconate was successful. With increased travelling to tropical and subtropical countries a rising incidence of tropical infectious diseases can be expected in Iceland. PMID- 19679923 TI - [Ampicillin resistant enterococci at Landspitalinn University Hospital and antimicrobial susceptibilities of enterococci in Iceland.]. AB - Hospital acquired infections caused by enterococci are an increasing problem, due to an increased number of infections and increasing bacterial resistance to antibiotics. During 1994 ampicillin resistant enterococci were discovered in specimens from three patients in one ward over a short time period. The patients were isolated and stool cultures were taken from everyone in the ward for selective culture for enterococci and subsequent antimicrobial susceptibility tests. Additional screening cultures were taken from patients in the intensive care unit, the oncology ward, one surgical and one paediatric ward. Cultures were also taken from the hospital sewage system. Antibiotic susceptibility of enterococci isolated from urine samples submitted to the Microbiology Department, Landspitalinn, during 1994 and 1995 were reviewed. In the index ward, specimens were obtained from 30 individuals for culture. One additional patient and one staff member were found to be colonised with ampicillin resistant enterococci. In the other wards a total of 23 samples were taken from selected patients for culture, but none of these cultures yielded ampicillin resistant enterococci. No ampicillin resistant enterococci were found in the sewage system of the hospital. Of a total of 41,181 urine specimens cultured at the Microbiology Department, 1,513 contained enterococci of which five were resistant to ampicillin (0.3%, all from 1994). We conclude that ampicillin resistant enterococci have not become established at Landspitalinn. It may be difficult to maintain a susceptible enterococcal population, however isolation of carriers and sensible use of broad spectrum antibiotics are likely to delay the establishment of multiresistant enterococci in Iceland. PMID- 19679924 TI - [Bronchial asthma and respiratory symptoms among Icelanders 20-44 years of age.]. AB - OBJECTIVE: To estimate the prevalence of asthma and respiratory symptoms in an urban population. MATERIAL: Eight hundred men and women aged 20-44 years, living in the capital Reykjavik and suburbs. METHODS: Participants answered a questionnaire, underwent skin prick testing for atopy, spirometry and a test for bronchial hyperresponsiveness (BHR) by methacholine challenge. RESULTS: There was 77% attendance. Altogether 16.6% reported wheezing or whistling at any time in the last 12 months. Altogether 32 (5.6%)answered yes to "Have you ever had asthma?" and the diagnosis had been confirmed by a doctor in all but four. Fourteen (2.5%) had suffered from an attack of asthma in the last 12 months wheras only 0.9% were currently using anti asthmatic drugs. BHR was found among 8.7% and atopy on skin testing among 20.5%. BHR was more common among those with airflow obstruction and three times more common among the atopic participants (18% vs. 6%, p%lt;0.01). By using a history of wheezing during the last 12 months together with BHR and/or a history of doctor confirmed asthma the prevalence of current asthma was found to be 5% in our sample. The main predictive factors for asthma were a history of breathlessness and nighttime breathing symptoms, but also atopy, airflow obstruction and a maternal history of asthma. CONCLUSION: Even by using a conservative defination, asthma is a common disorder among 20-44 years old Icelanders whereas the use of asthma medication is rather uncommon in this population. PMID- 19679925 TI - [Dietary fish-oil supplementation increases survival in mice following Klebsiella pneumoniae infection.]. AB - INTRODUCTION: Epidemiological studies have shown that high intake of omega-3 fatty acids correlates with low incidence of various diseases such as cardiovascular diseases, asthma, diabetes mellitus and various auto-immune disorders. It may therefore be suggested that omega-3 fatty acids have substantial impact on the immune system. Studies of the effect of omega-3 fatty acids on survival in bacterial infections have however been contradicting. A Dutch study from 1991 showed increased survival in mice fed fish-oil following infection with Klebsiella pneumoniae. Because of the contradicting results the authors conducted a study with the hypothesis that fish-oil intake increases survival after severe Klebsiella pneumoniae infection. METHODS: Thirty mice were fed fish-oil enriched diet (10%), olive-oil enriched diet (10%) or standard chow diet. After six weeks the mice were injected intramuscularly with l.oxlO2 cfu of Klebsiella pneumoniae. The survival was measured at regular time intervals for 120 hours. RESULTS: After 56 hours, 93% of the mice fed fish-oil were alive and 68% and 40% of the mice fed olive-oil and standard chow respectively. The overall survival after 120 hours was 40% in the fish-oil group, 25% in the olive-oil group and 20% in the standard group. The survival after 120 hours of the mice fed the fish-oil enriched diet was significantly better when compared to the two other groups (p=0.0034). DISCUSSION: We conclude that fish-oil enriched diet increases survival of NMRI mice following infection with Klebsiella pneumoniae when compared to olive-oil supplementation or standard chaw. We therefore conclude that the difference in survival is probably based on the effect of omega 3 fatty acid on the immune system. The immunological pathway is still unknown and our results encourage further studies. PMID- 19679926 TI - [Out of hours service in the Icelandic primary health care sector: an observational study.]. AB - BACKGROUND: Decisive changes have occurred in out of hours service in recent years, prompted by technological and methodological progress, improved communications and public demand. Concurrently, physicians' leisure time has come under increasing pressure. AIMS: To examine the workload, duties and com notmitments of primary care physicians (GPs) resulting from out of hours service. Further, to assess the organisation and quality of out of hours service and possible differences between urban and rural areas. MATERIAL AND METHODS: A postal survey containing 58 questions was sent to all primary health care physicians holding posts for at least six months on March 1st 1996. Of a total of 143 GPs, answers were returned from 100 (70%). RESULTS: All GPs on call (100%) were equipped with radios, mobile phones or pagers. They estimated that in 95% of the cases they could be contacted within five minutes jn an emergency. In an emergency and bad weather conditions 82% of doctors in urban areas could attend their patients within 30 minutes, and 100% within 60 minutes. Similar figures for rural areas were 10% within 30 minutes and 18% within 60 minutes. In the least populated districts 84% of the practitioners had to be on call 14 days or more per month. Serious emergencies (involving special training such as cardiac resuscitation or tracheal intubation) were relatively rare, and GPs expressed the necessity for regular refreshing courses in such fields. CONCLUSIONS: Access to doctors is good in this country. Workload and on-call duties are significantly heavier here than abroad, to some extent owing to outdated organisation. Various possibilities exist for improvement in the quality and reliability of out of hours service, but to be effective they must be supplemented by professional counselling and support, and facilities for further training in these fields. PMID- 19679927 TI - [Burn injuries in children in Iceland between 1982-1995.]. AB - OBJECTIVE: To increase our knowledge of burn injuries in children in Iceland and to induce education and prevention in order to reduce the incidence of burn injuries among children. MATERIAL AND METHODS: Data was collected from hospital records of all children 15 years and younger admitted with burn injuries to the University Hospital of Iceland, Paediatric Department, from 1982-1995. RESULTS: There were 290 children admitted, 179 boys and 111 girls, sex ratio 1.6. Children four years and younger were 72.8%. Approximately 21 children were admitted annually. Seasonal variation was noted with most admittances in December. The times when the injuries occurred peaked at lunch and dinner times. Scalds was most common, hot water caused 45.8% of the burn injuries, most frequently due to bathwater (15.2%). Hot liquids caused 26.9%, most often caused by coffee-, tea- and cacao drinks (19.3%). Flames caused 12.4% of the burn injuries, fireworks 5.5% and hot object 5.2%. Most of these accidents occurred at home (81.4%). CONCLUSION: Children four years and younger are most susceptible for burn injuries. Hot water and liquids caused most of these burn injuries. Burn injuries are common in childhood. Our data provide basis for better prevention. PMID- 19679928 TI - [Operations for liver injury 1968-93. A review from the Reykjavik City Hospital.]. AB - OBJECTIVE: The purpose of the study was to disclose the operative experience with liver injuries at the Reykjavik City Hospital for the years 1968-1993 and compare the results to those reported by others. MATERIAL AND METHODS: Journals for all patients undergoing an operation for liver injury during the period were studied. Age and sex, cause of injury, condition on arrival at the hospital, additional injuries, length of the operation, number of transfusions, hospital stay, indications for surgery, type of surgery, complications and mortality were all noted. The abbreviated injury severity scale (AIS-90) was used to classify the liver injuries. The injury severity score (ISS) was calculated for each patient. RESULTS: There were 41 patients, 28 males and 13 females. The median age was 20 years (5-78) and one fourth were children under 10 years. Blunt trauma caused 84% of the injuries and traffic accident the most common cause. Seventeen patients (42.5%) were in shock (systolic BP under 90) on arrival and 12 (29%) were still in shock at the beginning of the operation. Each patient had on the average 2.22 additional injuries inside or outside the abdomen. The number of transfusions required, hospital stay and operative time were extremely variable the median being 1.4 liters, 100 minutes and 15 days respectively. The most common indication for laparatomy was shock or fluid in the abdomen as shown by ultrasound and signs of peritoneal irritation. The liver was bleeding at the time of surgery in 51.3% of cases. Bleeding could in most instances be controlled with sutures. Three patients underwent a major hepatic resection and all survived. One of the three had an associated vena cava injury. There were 20 major complications of which abdominal sepsis and renal failure were the most common. Seven patients died (17%) but only one of liver bleeding on the operating table. Others died from brain injury, chest injury or multiple organ failure. Seventy one per cent had minor or moderate injury to the liver (class I or II) while 29% had major or massive injury (class III or VI). Twenty three patients (56%) had injury severity score (ISS) under 16. One of these 23 patients died, an elderly man with cirrhosis and incurable carcinoma of the liver. Other patients who died had ISS of 41 or higher. In this series are only patients who underwent an operation. In recent years there has been a growing tendency to treat liver injury without operation. From 1988-93 seven of 15 patients with liver injury were treated without operation. CONCLUSIONS: Most of the results in this study are similar to those reported by others. The mortality is low when compared to authors who mostly have closed injuries in their series as we do. No patient with normal liver before the accident died unless he had ISS of 41 or higher. This fact along with the low mortality seem to indicate that treatment was of high standard. PMID- 19679929 TI - [Wada test. An investigation of language and memory functions in epileptic patients evaluated for temporal lobectomy.]. AB - INTRODUCTION: Intracarotid sodium amytal injection was introduced as a clinical investigation of epileptic patients by Juhn Wada around 1950. The Wada test causes a brief inhibition of cerebral functions of the anaesthetized hemisphere, thus allowing tests to be performed on the contralateral hemisphere. The test is widely used to lateralize language functions and to assess the risk of postoperative amnesia in epileptic patients evaluated for temporal lobectomy Subjects and methods: Five epileptic patients were investigated. Three patients had hippocampal sclerosis and two had a benign tumour in the amygdala region. The sodium amytal was first injected to the hemisphere with seizure onset. After the development of paralysis of the contralateral side of the body, language and memory functions of the non-anaestetized hemisphere were assessed. The test was then repeated for the other hemisphere. RESULTS: The left hemisphere was dominant for language in three patients. In one patient the right hemisphere was dominant for language and in another patient language was bilaterally represented. In the three patients with hippocampal sclerosis, verbal and nonverbal memory was worse on the side of the lesion. This difference was not as marked for the two patients with lesion in the amygdala region. Total memory score was worse on the side of the lesion in all five patients. DISCUSSION: In both right and left handed individuals language is usually located in the left hemisphere. When epileptic seizures, with onset in the left hemisphere, start early in life, the language function can be transferred to the right hemisphere. This is a likely explanation for the right hemisphere language dominance in one patient. In all patients total memory score was lower for the hemisphere with seizure onset. This is in agreement with the suggestion of a lateralizing value of the Wada test. PMID- 19679930 TI - [Treatment of schizophrenic patients with depot-neuroleptics.]. AB - During almost four decades neuroleptic drugs have been the main weapon in doctor's armamentarium against schizophrenia and some other major psychiatric disorders. In Iceland seven different depot-neuroleptics from three subgroups are used. The main indication for depot-neuroleptics is to insure that the patient gets the prescribed amount of medication and to prevent a relapse from noncompliance. Depot-neuroleptics have the same side effects as neuroleptics in general. The main focus in this paper is on the following side effects: Extrapyramidal symptoms, tardive dyskinesia, psychic symptoms and neuroleptic malignant-syndrome. PMID- 19679931 TI - [Dermatitis in cats and humans caused by Cheyletiella mites reported in Iceland.]. AB - Cheyletiella mites (Acarina) are ectoparasites that infest cats, dogs and rabbits in many countries of the world. Upon contact with infested animals the mites may temporarily produce grouped, erythematous macules on the skin of humans which rapidly develop a central, vesicular papule. These signs are most often found on the arms and the trunk. Recently these typical signs were observed on the skin of the members of two different Icelandic families which both kept a Persian cat. An examination for ectoparasites on the cats revealed that both were infested by Cheyletiellaparasitovorax. It is unknown how and when the parasite was transmitted to Iceland. PMID- 19679932 TI - [Hypnotics and tranquillisers. Patients' knowledge and attitudes.]. AB - OBJECTIVE: To elicit patients' views and attitudes towards the use of hypnotics and tranquillisers and their knowledge about such drugs and household remedies. MATERIAL AND METHODS: During 1986-1993 a total of 577 individuals in Egilsstaethir district (3029 inhabitants) had used sedatives and/or tranquillisers. Out of this group a stratified randomised sample composing 208 individuals, received a questionnaire with 22 or 24 questions about the use of tranquillisers and hypnotics. RESULTS: The mean age was 55.1 years for users of tranquillisers, 60.1 years for the users of hypnotics. Women were in majority in both groups, most of them were married and working outside the home. Users of tranquillisers were divided into two main categories i.e. daily users and occasional users. Only a few used tranquillisers two or three times daily. Many used tranquillisers only as sleeping medicine. A fourth of the participants began using hypnotics after sleeping problems of less than one month. Doctors usually prescribed the medicines the first time they were used. 55% have tried household remedies against anxiety and sleeping problems. A majority had tried to stop using these medicines. Feelings of guilt because of the use of medicines was not common. Participants were well informed about how much sleep is needed. CONCLUSION: Our results show that most of the patients in the research area had gripped their doctors' message regarding sleeping problems, health promotion and the risk of addiction after continuous use of hypnotics and tranquillisers. It is however likely, that more education in this direction can further increase health promotion and minimise the use of drugs. PMID- 19679933 TI - [Non-convulsive status epilepticus. A rare but important reason for prolonged loss of conciousness.]. AB - Non-convulsive status epilepticus is a rare form of epilepsy. The predominant clinical feature is prolonged loss of consciousness without prominent motor features. The diagnosis is often difficult because of the non-specific nature of the symptoms and this diagnostic possibility has to be born in mind when patients present with unexplained alteration in the level of consciousness, especially if there is a prior history of epilepsy. The electroencephalogram plays a key role in the diagnosis and intravenous administration of diazepam is a useful diagnostic test, especially in conjunction with EEG. We present three patients recently diagnosed in Iceland. PMID- 19679934 TI - [Persistent organochlorines in Icelandic breast milk.]. AB - Persistent organochlorines are passed from a mother to her offspring and can affect his or her growth and development. The levels of persistent organochlorines in breast milk from 22 Icelandic women (18 primiparae and four bisparae) were investigated during the summer of 1993. The samples were collected at the Maternity Department of Landspitalinn, the University Hospital in Reykjavik. Dichloro-diphenyl-dichlorethene (DDE), hexachlorobenzene (HCB) and polychlorinated biphenyls (PCBs), were found in all samples but not hexachlorocyclohexane (HCH). No statistical difference was found between primi- and bisparae mothers. The levels found were similar to what has been reported for breast milk in neighbouring countries in recent years and lower than was found in France, Germany, Italy and among Inuits of arctic Canada but higher than was found in mothers from South-Canada and Great Britain. The levels, however, were well below estimated risk levels for infants. PMID- 19679935 TI - [Toxicity of monkshood. Review.]. AB - Monkshood, Aconitum napellus L. (Ranunculaceae), is considered one of the most poisonous plants growing in Europe. Monkshood and other Aconitum species are still used in Oriental and homeopathic medicine as analgesics, febrifuges and hypotensives. The neurotoxin aconitine is the principal alkaloid in most subspecies of monkshood. A review is presented, which includes historical aspects of monkshood as a poisonous and medicinal plant, the mode of action of aconitine, symptoms of toxicity, treatment and reports of recent poisoning incidents. In addition, results of quantitative HPLC examination of hypogeous and epigeous organs from a population of A. napellus ssp. vulgare cultivated in Iceland are discussed. The fact that children in Iceland have commonly been known to eat the sweet tasting nectaries in monkshood prompted an investigation of the alkaloidal content of these organs specifically. The low aconitine content found in the nectaries as well as in whole flowers accords with the absence of reported toxicity arising from the handling of flowers and consumption of nectaries from A. napellus in this country. PMID- 19679936 TI - Ionization of 4,4'-bis(phenylethynyl) anthracene by electron impact. AB - Ionization of a 4,4'-bis(phenylethynyl)anthracene (C(30)H(18), BPEA) molecule is studied for the first time at different energies of bombarding electrons in crossed electron and molecular beams. The relative cross-section of single ionization of a BPEA molecule in the energy range of 5-55 eV is measured. The ionization potential, E(I) = 7.62 +/- 0.2 eV, is determined using the threshold region of C(30)H(18)(+) ion yield energy dependence. A scheme of the BPEA molecule fragmentation is proposed. Experimental results are in reasonable accordance with calculations made in the framework of the density functional theory. PMID- 19679937 TI - Use of Nafion as a membrane separator in membrane introduction of mass spectrometry. AB - Nafion is a commercially available perfluorosulphonate cation exchange membrane commonly used as a perm-selective separator in chlor-alkali electrolysers and as the electrolyte in solid polymer fuel cells. In our experiments, a Nafion sheet membrane serves as the interface between the aqueous sample and the vacuum in membrane introduction to the mass spectrometer (MIMS). The penetration by volatile polar compounds (VOC-methanol, ethanol, 1-propanol), volatile non-polar compounds (VOC-benzene, toluene and p- xylene), semi-volatile low polar compounds (SVOC-fluorobenzene, chlorobenzene and bromobenzene) and non-volatile polar compounds (o-chlorophenol, m-chlorophenol and p-chlorophenol) in aqueous solution through the Nafion membrane to the mass spectrometer was studied. In all cases, a simple fragmentation pattern of the intact molecule was observed, typically with m/z = nominal mass + 1 as the most intensive ion current, which suggests that the ionisation process takes part in which water acts as the chemical ionisation reagent. No additional gases were needed for chemical ionisation. We also measured detection limits and linear dynamic ranges of all observed compounds with Nafion membrane MIMS. The observed detection limits were in the order of ppb for the alcohol and aromatic groups and for the halogenbenzene and monochlorophenol groups they were in the order of ppm. Linear dynamic ranges for all tested compounds were one order of magnitude. PMID- 19679938 TI - Gas chromatography electron ionization mass spectrometric analysis of O-alkyl methylphosphinates for verification of Chemical Weapons Convention. AB - We describe the gas chromatography/mass spectrometric (GC/MS) analysis of O-alkyl methylphosphinates (AMPs), which are included in schedule 2B4 chemicals in the Chemical Weapons Convention (CWC). GC/MS analysis of variety of AMPs and their deuterated analogues revealed that their fragmentations were determined by alpha cleavages, McLafferty +1 and hydrogen rearrangements. Based on the obtained electron ionization mass spectra of AMPs the fragmentation routes were rationalized, which were substantiated by the GC/MS analysis of deuterated analogues. PMID- 19679939 TI - "Soft"or "hard" ionisation? Investigation of metastable gas temperature effect on direct analysis in real-time analysis of Voriconazole. AB - The performance of the direct analysis in real-time (DART) technique was evaluated across a range of metastable gas temperatures for a pharmaceutical compound, Voriconazole, in order to investigate the effect of metastable gas temperature on molecular ion intensity and fragmentation. The DART source has been used to analyse a range of analytes and from a range of matrices including drugs in solid tablet form and preparations, active ingredients in ointment, naturally occurring plant alkaloids, flavours and fragrances, from thin layer chromatography (TLC) plates, melting point tubes and biological matrices including hair, urine and blood. The advantages of this technique include rapid analysis time (as little as 5 s), a reduction in sample preparation requirements, elimination of mobile phase requirement and analysis of samples not typically amenable to atmospheric pressure ionisation (API) techniques. This technology has therefore been proposed as an everyday tool for identification of components in crude organic reaction mixtures. PMID- 19679940 TI - Negative ion electrospray ionization mass spectrometry and computational studies on substituted 7-hydroxycoumarins. AB - Twenty-two substituted 7-hydroxycoumarins were studied by negative ion electrospray ionization collision- induced dissociation (CID) mass spectrometry. Fragmentation pathways were also investigated by computation method using the B3LYP density functional theory. In general, the most important fragmentations of the 7- hydroxycoumarin [M - H](-) ions were the elimination of CO(2) and CO which agreed with the calculated energies of the proposed fragmentation reactions. In most cases, methyl group elimination was also favorable. Methyl group elimination occurred in three different ways, the most interesting being hydrogen rearrangement from a neighboring alkyl group to a ring carbon, which led to a benzyl radical formation. In some cases, CH(2)CO elimination was observed as well. Isomeric compounds gave rise to different CID spectra. PMID- 19679941 TI - Structural characterization of nitrated 2'-hydroxychalcones by electrospray ionization tandem mass spectrometry. AB - Isomeric 2'-hydroxychalcones bearing nitro and methoxy groups in different positions of their skeleton were analyzed by tandem mass spectrometry (MS/MS) with electrospray ionization (ESI), in positive mode. Collision-induced dissociation of the protonated molecules, [M + H](+), formed under electrospray conditions were studied and it was found that the product ion spectra of these chalcones presented different fragmentation patterns depending on the position of the substituents on the molecule. The product ion spectra (ESI- MS/MS) of the B ring ortho-nitro substituted 2'-hydroxychalcone and of the 4'-methoxychalcones showed loss of OH, 2OH and combined losses of OH and H(2)O. These fragment ions were absent in the spectra of the respective meta- and para isomers. The observed differences in the product ion spectra of these nitrochalcones allowed identification of the o-nitro derivatives. Distinction between the meta- and para derivatives was not achieved. Chalcones bearing 6'-methoxy substituents showed distinct fragmentation from the one observed for their isomers, 4' methoxychalcones, since they present only one fragment ion, a typical ((0,alpha)A - H)(+) and, therefore, do not allow detailed structural information to be obtained, nor to differentiate between the o-, m- or p-nitro isomers. Overall, it was found that small changes in the substitution pattern of chalcones change their fragmentation considerably in the ESI-MS/MS, and that these features permit the differentiation of specific isomers of these 2'-hydroxynitrochalcones. PMID- 19679942 TI - Evidence of structure-selective fragmentations in the tandem mass spectra of protonated hydroxyalkylamino-1,4-naphthoquinones formed by electrospray ionisation. AB - The collision-induced dissociation of protonated hydroxyalkylamino-1,4 naphthoquinones depends strongly on the structure of the substituent [NHCH(2)(CH(2))(n)OH, n = 1-5; or NHCH(2)CH(CH(3))OH] on the quinone ring. Protonated naphthoquinones with an unbranched hydroxypropylamino side chain (n = 3) undergo facile and characteristic CH(2)O loss, whereas isomeric [M + H](+) ions with a branched hydroxypropylamino side chain do not. When n = 1, CH(2)O elimination occurs less readily, accompanied by CH(3)N loss, thus allowing this shorter side chain to be identified. Higher homologous species (n = 3-5) do not expel CH(2)O, but instead eliminate C(n + 1)H(2n)O, C(n + 1)H(2n + 2)O and (for n = 5) C(2n + 1)H(2n + 1)O. PMID- 19679943 TI - Electrospray ionization mass spectrometry as a valuable tool in the characterization of novel primaquine peptidomimetic derivatives. AB - Novel primaquine-derived antimalarials have been extensively characterized by electrospray ionization-ion trap mass spectrometry (ESI-MS). Experiments by in source collision-induced dissociation (CID) in the nozzle- skimmer region (NSR) or by tandem-MS are shown to be most valuable tools for the physicochemical characterization of these 8-aminoquinolinic drugs that also bear the biologically relevant imidazolidin-4-one scaffold. It was possible to find parallelism between compound stability in the NSR and its reactivity towards hydrolysis at physiological pH and T. Moreover, tandem-MS fragmentation patterns were characteristic for each family, providing a means for structural distinction of isomers and allowing to find interesting correlations between the relative abundance of particular fragments and relevant structure-activity determinants, such as Charton steric parameter, v. In conclusion, this work provides solid grounds to establish ESI-MS as a key tool for the physicochemical characterization of biopharmaceuticals bearing the 8-aminoquinoline and/or the imidazolidin-4-one moieties. PMID- 19679944 TI - Characterization of recombinant protein mutants by top-down sequencing using quadrupole time-of-flight mass spectrometry. AB - Top-down sequencing using quadrupole time-of-flight mass spectrometry is used as a direct way of locating the mutated sites of recombinant proteins and posttranslational modification in a protein. Several mutants of barstar, expressed in E.coli, were confirmed by analyzing the fragmentation pattern of mutants. A contaminant protein, that appeared while purifying mutants of barstar, was identified as acyl carrier protein from E.coli with a posttranslational modification on serine residue, indicating that the protein was biologically active. A mutant of ribosomal protein S6 has been characterized with neutral loss of ammonia at the N-terminal region of the protein. The power of the "top-down" approach in characterizing the mutants of recombinant proteins has been demonstrated. PMID- 19679945 TI - An electrospray ionization mass spectrometric study of iron binding to amyloid beta peptides. AB - Iron and other metal ions appear to play an important role in protein aggregation and are therefore likely to provide a link between protein aggregation and oxidative damage. This work reports on iron binding to amyloid- beta peptide (Abeta1-40), which affords a very specific electrospray ionization mass spectrometric (MS) spectrum. Both MS and MS/MS study confirmed that amyloid-beta peptide displays a high affinity toward iron(III) ions, producing multi charged molecular ions and peptide aggregates. Finally, the circular dichroism spectra indicate an unexpected modification of Abeta1-40 peptide conformation upon iron binding. PMID- 19679946 TI - Probing the mechanism of action of potential anticancer agents at a molecular level using electrospray ionisation Fourier transform ion cyclotron resonance mass spectrometry. AB - Treating cancer without harming healthy tissue is an important goal in modern medicine. Our research group has developed a series of novel, relatively non toxic glycosidic prodrugs that are activated to give the corresponding highly cytotoxic drugs selectively in the tumour tissue. Our first investigations have shown a high duplex DNA alkylation efficiency of the drugs, whereas the prodrugs showed almost no tendency for alkylation of duplex DNA. Herein we report on novel investigations of the mode of action of the anti-cancer drugs on a molecular level. Using high-resolution mass spectrometry, we determined the reactivity of these drugs as well as of other drugs of similar structure against different nucleophiles such as RNA and the tripeptide glutathione. In addition, the new drugs were also tested for their interaction with duplex DNA. All compounds show a high reactivity against duplex DNA, whereas the alkylation efficiency regarding RNA and glutathione is only poor. Furthermore, the alkylation of duplex DNA correlates qualitatively but not quantitatively with the cytotoxicity of the drugs. Consequently, other factors besides the alkylation efficiency such as the stability of the drugs seem to influence their biological activity. Altogether the results show that high-resolution mass spectrometry constitutes a powerful method for studying the mode of action of drugs on a molecular level. PMID- 19679947 TI - Approach to diagnosis of auto-immune hepatitis. AB - Auto-immune hepatitis (AIH) is one of the chronic liver diseases, seen predominantly in women, resulting from dysregulated immune mechanisms not yet clearly defined. Based on a combination of clinical and laboratory parameters with both positive and negative weights, the International AIH Group devised a scoring system in 1993. The system was modified in 1999 and has proven useful for both diagnostic and research purposes. This review deliberates on the clinical, immunological and histological features of this entity. PMID- 19679948 TI - Making and using inexpensive manually constructed tissue micro-array: experience of a tertiary care hospital in India. AB - BACKGROUND: Tissue micro-array enables the analysis of a large number of tissues simultaneously. Widespread use of this technology is hampered by the high cost of commercial array instruments. We describe our experience of constructing tissue micro-array in a simple method using easily available and inexpensive instruments. MATERIALS AND METHODS: We used an 11-19 gauge (G) bone marrow trephine biopsy needle/ small sized slotted screwdriver to punch holes in the wax blocks. Cores were taken from donor tissue blocks using a bone marrow trephine biopsy needle and arrayed into host paraffin wax blocks. A detailed database was constructed for each array constructed. RESULTS: The array blocks were used over a period of one year as internal control for immunohistochemistry (IHC), quality control and research. It took about 10 minutes to construct a nine-dot array and about one hour for a 56-dot array. During IHC, the average loss of control dots was less than one per cent. We did not see any loss of antigenicity in the control sections even after four weeks storage. DISCUSSION: Tissue array construction by the technique described here is inexpensive and reliable alternative to automated instruments. Because it is easy to modify the arrays by varying the core size, it is easy to adapt this to individual labs and requirements. We recommend using blocks with cores in 3 x 3 to 5 x 4 grids as controls in IHC and for standardizing antibodies and array blocks with a larger number of cores for research. PMID- 19679949 TI - Significance of deep section in small gastric biopsies. AB - AIM OF STUDY: Chronic gastritis is defined as the presence of chronic mucosal inflammatory changes leading eventually to mucosal atrophy and epithelial metaplasia. This condition constitutes a background for dysplasia and thereby carcinoma. Detection of exact histopathology of inflammatory process is necessary in biopsy specimen. We designed the current study to determine the value of taking more sections in small gastric biopsies for better histopathologic evaluation. MATERIALS AND METHODS: Gastric biopsy specimen of children who suffered from gastrointestinal (GI) symptoms was sent in 10% formalin to our laboratory. After routine processing, three slides with several sections on them were taken from the specimen: t0 he first was named the superficial section, the second was stained by Giemsa and the third was named deep section (further sections after this slide will diminish in size). The slides were not taken exactly consecutively but several sections were discarded between them. The purpose of this study is to compare the superficial and deep sections for detection of inflammatory processes. RESULTS: In 1062 specimens the results of superficial section and deep section were the same (87.1%) and in 158 specimens the results were different. In 88 (7.2%) specimens deep section was diagnostic. The difference was seen usually as normal tissue in superficial sections but presence of lymphoid follicle in deep sections. The difference between superficial and deep sections was statistically significant. Although obtaining more sections will put an economic burden on the laboratory, we propose that in small gastric biopsies, it is helpful in better evaluation of histopathological changes. PMID- 19679950 TI - Primary hyperparathyroidism: a clinicopathological experience. AB - BACKGROUND: Parathyroid neoplasms form a small percentage of head and neck neoplasms. Primary hyperparathyroidism is caused by parathyroid adenomas, hyperplasia or, rarely, a carcinoma. MATERIALS AND METHODS: This is a retrospective study of 48 parathyroidectomies received in the Department of Pathology of a major teaching hospital over a period of 10 years. Clinical, biochemical and radiological details were retrieved from medical records. Information regarding routine gross and microscopic examination findings (including frozen section and paraffin sections) was retrieved from departmental records. RESULTS: We had 43 adenomas, three hyperplasias and two carcinomas. The most common age group was 21-30 years. The female:male ratio was 2.5:1. Most patients presented with skeletal manifestations. The pre-operative diagnosis was assisted by ultrasonography in 11 cases, computerized tomography of the neck in 10 cases and sestamibi scans in three cases. Intra-operative parathormone monitoring was performed in one case of adenoma. Frozen section was requested in 28 of 48 cases. There was a discrepancy between frozen section diagnosis and paraffin section diagnosis in two cases. CONCLUSION: Histopathologic diagnosis is an important guide to decide the type of surgical management. Although pathologic features of parathyroid carcinoma are diagnostically reliable, those of the more commonly encountered lesions of adenoma and hyperplasia may be overlapping and, therefore, indistinctive, more so if only a single gland is available for examination. Because parathyroid lesions are only occasionally encountered by the surgical pathologist, awareness of the spectrum of histologic features along with knowledge of recent trends in diagnosis and surgical management are important. PMID- 19679951 TI - Histopathological study of pruritic papular eruptions in HIV-infected patients in relationship with CD4, CD8 counts. AB - BACKGROUND: Pruritic papular eruptions (PPE) are common cutaneous manifestations in HIV-infected patients. Their frequencies, patterns and associated factors vary from region to region. There is no clear consensus on the etiology of PPE, the exact spectrum of the condition, the pathologic findings, or treatment. The present study is aimed at documenting the histopathological patterns of PPE, and their relation with CD4, CD8 counts in HIV-infected patients. MATERIALS AND METHODS: The present study lasted for 12 months (September 2005 to August 2006). After informed consent, data on skin disorders, HIV status, CD4 and CD8 counts were obtained by physical examination and laboratory methods. RESULTS: Of the 36 HIV-infected patients with clinical diagnosis of PPE, the most common histopathological patterns were prurigo simplex and eosinophilic folliculitis. The mean CD4 count of PPE was 186.49 cells/mm3 with mean CD8 count of 619.60 cells/ mm3, and the CD4:CD8 ratio was 0.32. Patients with prurigo simplex had significantly lower mean CD4 counts (50.36 +/- 30.67 cells/cmm 3 , P < 0.001) while in eosinophilic folliculitis, the mean CD8 counts were higher (1239.77 +/- 402.30 cells/cmm3). CONCLUSION: We conclude that histopathology helps in specifying the pattern of PPE and also indicates underlying immunosuppression and can be a marker of advanced HIV infection. Thus, correlation between the histopathology and immunology findings helps to know the disease process. PMID- 19679952 TI - In-house daily consensus conference: an important quality control/quality assurance activity--experience at a major referral center. AB - BACKGROUND: For every practicing histopathologist, improvement of diagnostic accuracy is an important objective. Personal consults are an important component of quality control (QC)/quality assurance (QA) in our Section of Histopathology. In addition, the College of American Pathologists recommends a daily in-house consensus conference, which is a prospective system by which all difficult and problematic cases are reviewed and discussed and signed out by consensus. DESIGN: In-house consensus conference is held daily using a multi-headed microscope. This collegial session is run by the seniormost consultant in the section and is attended by all histopathology consultants and residents. The consultants and residents present cases of their choice for discussion. The cases may be selected due to diagnostic difficulty, unusual nature of a case, management purposes such as performance of additional biopsies, special studies, etc., or request on the part of clinician or patient. Cases may be shown once or, in case of lack of consensus or difficulty in diagnosis, more than once after additional work-up suggested by the conference. RESULTS: In a 4-month period, 774 (4.1%) cases of a total of over 14,000 well-mixed surgical cases were brought to the in-house daily consultation conference. Four hundred ninety-three cases (63.7%) were conclusively decided the first time while 198 cases (25.5%) were decided by consensus after being shown twice. In 83 cases (10.7%), a definite diagnosis could not be given. The cases on which a definite diagnosis was not possible represents 0.59% of all cases received in the department during the study period. The most common cases were shown from the gastrointestinal tract (115 cases or 14.8%), lymph nodes (110 cases or 14.2%) and soft tissue (82 cases or 10.6%). In most cases in which a definite diagnosis could not be given, the main reason was scanty material or crushed nature of the tissue. CONCLUSION: The in-house daily consensus conference is an extremely useful QC/QA exercise, which is very important in reaching an accurate diagnosis in difficult and challenging cases and minimizing diagnostic errors. PMID- 19679953 TI - Significance of cytomorphological and microbiological examination of bile collected by endoscopic cannulation of the papilla of vater. AB - BACKGROUND: Bile analysis yields important information such as "biliary microlithiasis" and biliary tract colonization by microorganisms like Salmonella typhi or paratyphi, Escherichia coli, etc., which may progresses to cholelithiasis and have been found to have a role in the development of gallbladder cancer in India. AIM: To perform microscopic, cytomorphological and microbiological examination of bile collected during endoscopic retrograde cholangiopancreatography in patients with benign and malignant lesions of the gallbladder. MATERIALS AND METHODS: Bile was collected from 48 patients by a catheter inserted through the cystic duct during endoscopic retrograde cholangiopancreatography. Direct microscopy and grading of crystals was performed. Smears prepared from centrifuged deposits were stained with Giemsa, Papanicolaou stain and Gram's stain for cytological and microbiological examination. Special staining for Helicobacter pylori was performed using Loeffler's Methylene blue and Warthin Starry stain. The rest of the sample was used for culture and complete microbiological examination. RESULTS: Thirty-six patients had inflammatory lesions while 12 had malignant lesions. Crystals were observed in 93% of the cases, 13 (28.8%) had only cholesterol crystals, three (6.6%) had bilirubinate and 29 (64.4%) had both cholesterol and bilirubinate crystals. Smears from the centrifuged deposit mainly showed coccoid or cocobacillary bacteria on Gram's staining (81.3%). Five of 12 (41.6%) malignant cases showed epithelial atypia while none of the benign or inflammatory lesions showed such a change in hematoxylin and eosin and Pap-stained smears. Microbiologic analysis showed Staphylococcus aureus (14%), S. saprophyticus (5.5%), Peptococcus (5.5%), Peptostreptococcus (5.5%), Proteus mirabilis (5.5%), E. coli (17%), Enteorbacter cloacae (5.5%) and H. pylori (2.8%). CONCLUSION: Complete microscopic and microbiological examination of bile directly obtained from the common bile duct during endoscopic retrograde cholangiopancreatography yielded good results as fresh samples could be analyzed. It may help in diagnosis of many gallbladder lesions in early stages, thereby reducing the morbidity and mortality. PMID- 19679954 TI - Bone marrow biopsy in non-Hodgkin lymphoma: a morphological study. AB - CONTEXT: Bone marrow (BM) biopsy is an integral part of staging work-up for non Hodgkin lymphoma (NHL). AIMS: To study the characteristics of BM involvement in NHL with respect to incidence, histologic pattern and morphology of infiltration and its discordance with the histology of primary anatomic site. SETTINGS AND DESIGN: Forty-nine cases of NHL in which BM biopsy was performed for staging were included in this study, the primary site being classified according to the WHO classification for NHL. MATERIALS AND METHODS: A prospective study of 49 cases was conducted. Bilateral BM biopsy was obtained from the posterior superior iliac spine. The biopsies were fixed in 10% buffered formalin solution and decalcified using 10% formal - formic acid for 4 - 6 h followed by routine processing. The serial sections were stained by hematoxylin and eosin and reticulin stains. RESULTS: BM biopsy showed involvement by lymphoma in 27 cases (55.10%). Unilateral positivity was found in four cases (14.81% cases). The overall incidence of marrow involvement by NHL was 55.1%. The incidence of involvement was higher in T-cell lymphomas when compared with B-cell lymphomas and predominant pattern of involvement was mixed. Diffuse large B-cell lymphomas had the lowest incidence in all the B-cell lymphomas. A discordant histology between BM and primary anatomic site was found in 29.63% (8/27) of the cases, where it was seen more in follicular lymphomas and diffuse large B-cell lymphomas. CONCLUSIONS: Critical examination of BM biopsies can increase the diagnostic accuracy, thereby contributing to the prognosis and appropriate treatment modalities. PMID- 19679955 TI - Ventilator-associated pneumonia caused by carbapenem-resistant Enterobacteriaceae carrying multiple metallo-beta-lactamase genes. AB - CONTEXT: Ventilator-associated pneumonia (VAP) is a leading nosocomial infection in the intensive care unit (ICU). Members of Enterobacteriaceae are the most common causative agents and carbapenems are the most commonly used antibiotics. Metallo-beta-lactamase (MBL) production leading to treatment failure may go unnoticed by routine disc diffusion susceptibility testing. Moreover, there is not much information on association of MBL-producing Enterobacteriaceae with ICU acquired VAP. Therefore, a study was undertaken to find out the association of MBL-producing Enterobacteriaceae with VAP. SETTINGS: This study was conducted in a large tertiary care hospital of North India with an eight-bed critical care unit. MATERIALS AND METHODS: The respiratory samples (bronchoalveolar lavage, protected brush catheter specimens and endotracheal or transtracheal aspirates) obtained from VAP patients (during January 2005-December 2006) were processed, isolated bacteria identified and their antibiotic susceptibilities tested as per standard protocols. The isolates of Enterobacteriaceae resistant to carbapenem were subjected to phenotypic and genotypic tests for the detection of MBLs. RESULTS: Twelve of 64 isolates of Enterobacteriaceae were detected as MBL producers, bla(IMP) being the most prevalent gene. Additionally, in three strains, simultaneous coexistence of multiple MBL genes was detected. CONCLUSION: The coexistence of multiple MBL genes in Enterobacteriaceae is an alarming situation. As MBL genes are associated with integrons that can be embedded in transposons, which in turn can be accommodated on plasmids thereby resulting in a highly mobile genetic apparatus, the further spread of these genes in different pathogens is likely to occur. PMID- 19679956 TI - Direct drug susceptibility testing of Mycobacterium tuberculosis to primary anti tubercular drugs by nitrate reductase assay. AB - OBJECTIVES: Traditional drug susceptibility testing for Mycobacterium tuberculosis takes weeks and/or expensive. In this study, we evaluated nitrate reductase assay for drug susceptibility testing which is faster than the visual detection of colonies. MATERIALS AND METHODS: 32 clinical specimens (direct microscopy positive for AFB with 1+, 2+ or 3+ grading) were decontaminated and the sediment was inoculated onto the L-J medium with INH or Rifampicin incorporated with Potassium nitrate and the same medium without antibiotics at 1;10 dilution as control. After 2 weeks, the control was first tested for color change with addition of nitrate reductase reagents. If found positive, the media with antibiotics were tested and compared. Futher incubation was done if the control was found to be negative. The results obtained was compared with standard direct proportion method for drug susceptibility testing. RESULTS: Resistance of isolates as shown by both methods for INH and Rifampicin was 37.5% and 31.3% respectively. The results showed that NRA and proportion method do not differ significantly ( P < 0.05 for both drugs). Thus an excellent agreement between the results of NRA and proportion method was found for two primary anti-tubercular drugs, 87.5% for INH and 97% for Rifampicin. CONCLUSION: Nitrate reductase assay is a rapid and inexpensive method for susceptibility testing of M. tuberculosis for primary anti-tubercular drugs and could be an alternative to existing methods, particularly in resource poor settings. PMID- 19679957 TI - Diagnostic efficacy of C4d immunostaining in the detection of the humoral component of renal allograft rejection and therapeutic implications. AB - CONTEXT: In the current scenario of renal transplantation, the role of immunological methods in the detection of C4d has emerged as a useful adjunct in the recognition of acute humoral rejection (AHR). Few reports of this nature are available from the Indian context although there are several from the Western literature. AIMS: To study the humoral component of renal allograft rejection in patients presenting clinically with graft dysfunction by histopathological detection of polymorphs in the peritubular capillaries and the expression of C4d using immunological techniques, as well as the response of patients to appropriate antirejection therapy. SETTINGS AND DESIGN: This study from a tertiary care center reemphasizes the importance of recognition of AHR as a cause of renal allograft dysfunction. MATERIALS AND METHODS: Percutaneous renal biopsies were obtained from 40 postrenal transplant patients and evaluated for C4d using immunofluorescence and immunohistochemical methods. STATISTICAL ANALYSIS USED: SPSS software. RESULTS: Positive expression of C4d was seen in a total of 19/40 cases (44.4%) indicating immunological evidence of AHR. Diffusely positive cases were treated with IV immunoglobulin therapy, plasmapheresis and Rituximab following which graft function was restored. Patients with minimal to focal positive expression of C4d responded well to pulse steroids and change in immunosuppressive therapy. CONCLUSIONS: C4d staining is a useful adjunct to routine histopathological methods in evaluating the humoral component of acute renal allograft dysfunction and helps in planning appropriate antirejection therapy with the goal of achieving long-term graft survival. PMID- 19679958 TI - Improved microscopical detection of acid-fast bacilli by the modified bleach method in lymphnode aspirates. AB - OBJECTIVES: To improve the smear microscopy for detection of acid-fast bacilli (AFB) in fine needle aspiration cytology (FNAC) of lymph node using the bleach method and also to compare this with cytological diagnosis and the conventional Ziehl-Neelsen (ZN) method. STUDY DESIGN: In 99 consecutive patients with clinical suspicion of tuberculosis (TB) presenting with lymphadenopathy, FNACs were performed. Smears from the aspirates were processed for routine cytology and the conventional ZN method. The remaining material in the needle hub and/or the syringe was used for the bleach method. The significance of the bleach method over the conventional ZN method and cytology was analyzed using the chi2 test. RESULTS: Of 99 aspirates, 93 were studied and the remaining six were excluded from the study due to diagnosis of malignancy in 4.04% (4/6) and inadequate aspiration in 2.02% (2/6). Among the 93 aspirates, 33.33% (31/93) were positive for AFB on conventional ZN method, 41.94% (39/93) were indicative of TB on cytology and the smear positivity increased to 63.44% (59/93) on bleach method. CONCLUSION: The bleach method is simple, inexpensive and potent disinfectant, also limiting the risk of laboratory-acquired infections. The implementation of the bleach method clearly improves microscopic detection and can be a useful contribution to routine cytology. PMID- 19679959 TI - Serological profile of HSV-2 in patients attending STI clinic: evaluation of diagnostic utility of HSV-2 IgM detection. AB - OBJECTIVE: The present study was done to evaluate the serological profile of herpes simplex virus-2 (HSV-2) among patients attending sexually transmitted infections (STI) clinic and to determine the utility of detecting HSV-2 IgM antibodies in such patients. A correlation of HSV-2 infection with other STI including HIV has also been attempted. MATERIALS AND METHODS: Hundred consecutive patients who attended STI clinic, with one or more of the complaints as enunciated by WHO in syndromic approach for the diagnosis of STI, were included as subjects. All subjects were screened for common STI by standard laboratory procedures/ commercially available kits. HSV-1 and HSV-2 IgM antibody was detected by commercially available enzyme immuno assay kit in all patient's sera. Sera were also tested for other STI, namely HIV, Hepatitis B virus, Hepatitis C virus and Treponema pallidum. Antigen detection for Chlamydia trachomatis was done in genital swabs of all patients by Bio-Rad Chlamydia Microplate EIA 31189 (United States) kit. RESULTS: Thirty patients were found to have genital herpes. In 17/30 (56.6%) patients, HSV-2 serology was found to correlate with the clinical diagnosis. The coexistence of other infection in HSV-2 seropositive patients was detected in 8/30 patients. None of the patients having concomitant infections were clinically diagnosed accurately. Sensitivity, specificity, positive predictive value and negative predictive value of IgM antibodies for the diagnosis of genital herpes was 73.91%, 90.91%, 70.83% and 92.91% respectively. CONCLUSION: HSV-2 IgM detection could only be used as a supportive test for the diagnosis of genital herpes . It needs to be emphasized that the sensitivity and positive predictive value scores are pointers for further improvement in the commercial assay systems and a large sample size may determine the broader utility of such systems. PMID- 19679960 TI - Melasma: a clinicopathological study of 43 cases. AB - Melasma is a symmetrical hypermelanosis of the exposed skin characterized by brown macules on the sun-exposed areas of the skin. The present study was carried out on 43 patients to analyze the correlation between histopathological features with clinical and Wood's light examination. The study showed a clinical and histopathological discordance of 16.2%. Solar elastosis (55.8%) was the single most common histological finding apart from increased melanin concentration, epidermal flattening and dermal lymphomononuclear inflammation. PMID- 19679961 TI - Hemoglobin color scale a diagnostic dilemma. AB - Hemoglobin color scale (HCS) is a commercially available test to screen anaemia in the absence of laboratory based hemoglobinometry. The present study was aimed at to compare the efficacy of HCS with Sahli's method (SM) for haemoglobin estimation and to estimate its sensitivity and specificity with respect to auto analyzer as the gold standard. The study was conducted from November 2006 to April 2007 at the department of hematology, All India Institute of Medical Sciences, New Delhi as a project of World Health Organization. The haemoglobin level was measured by all the three methods in 401 patients attending Haematology out patient department. Consent was taken from all the patients. Sensitivity of Sahli's method was 98.2% and specificity was 66.2%, whereas the sensitivity of HCS was 30% and specificity was 100%. Sahli's method was found to be in good agreement with autoanalyzer (gold standard). It was thus concluded that HCS is not as efficacious, as sahil's method for hemoglobin estimations in field. PMID- 19679962 TI - Osteopetrosis: a rare cause of anemia--review of literature. AB - Normocytic anaemia is caused either by hypoproliferation of haemopoietic tissue or increased destruction of red cell. Osteopetrosis is a rare cause of anaemia. The leading clinical features are pallor, growth failure, hepatosplenomegaly.On hematological examination, anemia, thrombocytopenia, leucocytosis and myelophthisic anemia are commonly observed in this disease. We are highlighting osteopetrosis as a rare cause of anemia presenting to us for evaluation of anemia. PMID- 19679963 TI - Extrarenal calyces: a rare anomaly of the renal collecting system. AB - The abnormalities of the renal collecting system represent a complex and often confusing subset of urological anomalies. They manifest in many ways and often make preoperative diagnosis difficult. Extrarenal calyces (wherein the calyces and renal pelvis lie outside the renal parenchyma) is one of the rare anomaly of the collecting system. This anomaly may be associated with other anomalies of the urogenital system. We describe in this case report an unusual case of extrarenal calyces with five long calyces draining a hydronephrotic kidney with associated ureteral atresia. Radiological investigations failed to clearly define these abnormalities. The present case helps in understanding the characteristics of a rare, complex anomaly of the urinary system. PMID- 19679964 TI - Fulminant amebic colitis: a study of six cases. AB - BACKGROUND: Amebic colitis although common, rarely presents as fulminant colitis which has a high morbidity and mortality unless treated promptly and appropriately. AIM: To study the clinical, morphological features and outcome of fulminant amebic colitis (FAC). MATERIALS AND METHODS: A retrospective study of six patients who underwent surgical resections from 2002-06 and were diagnosed with FAC, was carried out. The morphological features assessed included the average number of trophozoites per high-power field and the depth of invasion of trophozoites into the muscularis propria. RESULTS: The study included five adults and one child who underwent surgery for fulminant colitis. Interestingly, a definite preoperative diagnosis of amebic colitis was made only in one patient and suspected in another. Intraoperatively, multiple perforations of the intestine with peritonitis were the most common findings. Gross examination typically revealed multiple ulcers with exudate and intervening normal mucosa. Microscopically, ulceration and myonecrosis with trophozoites within the exudate were seen in all cases. Trophozoites invading the muscularis propria were seen in five cases. Of the cases that showed myoinvasion by trophozoites, two patients expired within two weeks of surgery. One of the patients who expired also showed co-infection with Actinomyces. CONCLUSION: FAC is an uncommon outcome in amebic colitis with a high mortality requiring prompt surgical intervention. PMID- 19679965 TI - Congenital neuroblastoma with liver metastasis presenting with Hashimoto Pritzker disease. AB - A two-month-old female child presented with discrete skin-colored papules on the forehead and scalp. She was earlier diagnosed to have neuroblastoma of the adrenal gland which had metastasized to the liver. Histological sections of the skin lesion showed a nodule composed of sheets of cells with abundant eosinophilic cytoplasm and an occasional grooving of the nucleus. These cells were CD 68 positive. Eosinophilic abscesses were seen in between the neoplastic cells. PMID- 19679966 TI - Osseous metaplasia and mature bone formation with extramedullary hematopoiesis in follicular adenoma of thyroid gland. AB - Follicular adenomas of the thyroid may be subjected to degenerative changes like hemorrhagic and cystic changes, fibrosis, and calcification. Mature bone formation is a rare phenomenon, but extramedullary hematopoiesis (EMH) has also been rarely reported in thyroid gland. The combination of mature bone formation and EMH is rarer and has been reported, in a single case report, in a multinodular goitre. We describe a case of follicular adenoma with histologically proven osseous metaplasia and mature bone formation with EMH in a middle- aged woman, which, to our knowledge, is the first case in English language literature. PMID- 19679967 TI - Oncocytic sialolipoma of the submandibular gland with sebaceous differentiation: a new pathological entity. AB - CASE REPORT: We report the case of an oncocytic sialolipoma of the submandibular gland with sebaceous differentiation in a 73-year-old man. The initial symptom was a right submandibular painless mass. Ultrasonography showed a hypoechoic oval mass posterior to the submandibular gland. The tumorectomy was performed with preservation of the salivary gland. The tumor was composed of mature adipose tissue surrounded by a thin fibrous capsule, multiple nodules of oncocytes, normal ductal-acinar units with focal ductal sebaceous differentiation. DISCUSSION: We reviewed literature of the reported cases of mixed tumors of the salivary glands composed of mature adipose tissue with oncocytosis, salivary ducts, and acini with sebaceous differentiation. CONCLUSIONS: Sialolipoma and lipoadenoma with or without oncocytosis and/or sebaceous differentiation should be considered organ-specific tumors with a distinct histological appearance and specific terminology. PMID- 19679968 TI - Frontal recurrence of medulloblastoma five years after excision and craniospinal irradiation. AB - Medulloblastomas were originally classified under gliomas of the cerebellum until Bailey and Cushing in 1925 named these tumors as medulloblastoma. At present these tumors are classified under primitive neuroectodermal tumor. Surgical excision followed by craniospinal irradiation is the treatment of choice. A 13 year-old-girl operated for posterior fossa medulloblastoma 5 years ago presented with history of headache and vomiting on and off for 4 days in late August 2008. The MRI showed left frontal tumor which on excision was reported as medulloblastoma. Even after optimal treatment reports of recurrence abound in literature. The most common location is in the posterior fossa, followed by spinal, supratentorial, and uncommonly, systemic metastases. We conclude that medulloblastomas are highly aggressive tumor with high local recurrences if the initial excision is incomplete and that recurrence in the supratentorial area although uncommon is still a possibility. This mandates regular follow up of these children till adulthood to catch early recurrences and metastatic disease. PMID- 19679969 TI - Angioleiomyoma of the nasal cavity. AB - Angioleiomyoma is a rare benign tumor in the nasal cavity. There are very few reports in literature. A 34 year old male presented with left sided nasal obstruction and epistaxis. Clinical evaluation revealed a lesion replacing the anterior two-third of the Left inferior turbinate. An endoscopic excision under hypotensive anesthesia was performed. Histopathology confirmed a diagnosis of Angioleiomyoma. The cause and site of origin of angioleiomyomas when they arise from the inferior turbinate remains unclear. We review the literature available on nasal angioleiomyoma. When limited to the nasal cavity endoscopic excision is the treatment of choice. PMID- 19679970 TI - Metanephric stromal tumor: a novel pediatric renal neoplasm. AB - Metanephric stromal tumor of kidney is a novel pediatric benign stromal specific renal neoplasm. A few cases have been reported in adults also. This tumor is usually centered in the renal medulla with a characteristic microscopic appearance which differentiates this lesion from congenital mesoblastic nephroma and clear cell sarcoma of the kidney. In most cases complete excision alone is curative. The differentiation of metanephric stromal tumor from clear cell sarcoma of the kidney will spare the child from the ill effects of adjuvant chemotherapy. In this communication we describe the gross and microscopic features of metanephric stromal tumor in a one-month-old child with good prognosis. PMID- 19679971 TI - Sclerosing hemangioma with lymph nodal metastases. AB - A case of sclerosing hemangioma of the lung is reported in a young male, who presented with recurrent cough and streaky hemoptysis for three years. The tumor was situated in the right upper lobe and was large (over 9 cm), multicentric and associated with metastases to the regional lymph nodes. To the best of our knowledge, only 14 cases with such metastases have been reported. PMID- 19679972 TI - Leydig cell tumor: an unusual presentation. AB - Leydig cell tumor is a benign tumor of the testis and malignant transformation, if present, is rare. The case presented here showed certain features of malignancy but no infiltration beyond the capsule or metastasis. The case could not be labeled as benign or malignant and patient is on follow-up. Differential diagnosis and clinical implications of a case in the borderline zone are discussed. PMID- 19679973 TI - Long segment ileal duplication with extensive gastric heterotopia. AB - Duplications of the alimentary tract are rare congenital anomalies which can be found at all levels of the alimentary tract. Majority of the duplications present as spherical cysts and usually range from a few millimeters to less than ten centimeters in size. Duplications produce complications such as intestinal obstruction or hemorrhage. A two-month-old infant presented with recurrent episodes of bleeding per rectum. Laparotomy revealed a giant ileal duplicated bowel segment which exhibited extensive gastric heterotopia with focal ulceration. PMID- 19679974 TI - Renal mucinous tubular and spindle cell carcinoma. AB - Mucinous tubular and spindle cell carcinoma is a rare variant of renal cell carcinoma, which has recently been described. It has a low malignant potential and is usually confined to the kidney. These are thought to be of the loop of Henle or distal nephron origin. We report a 65-year-old male who presented with flank pain, hematuria and a well-defined renal mass that was diagnosed as mucinous tubular and spindle cell tumor. PMID- 19679975 TI - Endodermal sinus tumor of the vagina in children: a report of two cases. AB - Malignant tumors of the vagina in infants and children are rare, with primary germ cell tumor (endodermal sinus tumor [EST]) being rarer, which carries a dismal prognosis. This tumor is often clinically mistaken as botryoid rhabdomyosarcoma and, on histopathological examination, is often misdiagnosed as clear cell adenocarcinoma. Two cases of EST of the vagina in infants aged 9 and 17 months are reported. Both the patients presented with bleeding per vagina and clinically and by ultrasonography were diagnosed as sarcoma botryoides. Both were diagnosed as primary yolk sac tumors. The serum alfa fetoprotein (AFP) level was 4325 and 9328 ng/ml, respectively. One patient aged 9 months expired 2 months after receiving chemotherapy. The other, aged 17 months, was given chemotherapy followed by surgery with reduction of the AFP levels. PMID- 19679976 TI - Choroid plexus carcinoma: report of two cases. AB - Choroid plexus carcinomas (CPCs) are rare malignant counterparts of choroid plexus papilloma which occur in infants and children with a predilection for the posterior fossa and have a poor prognosis. We report two cases of CPC diagnosed in a 5-year-old boy and a 12-year-old boy and discuss the clinicopathologic features. PMID- 19679977 TI - Serous microcystic adenoma of the pancreas. AB - Serous microcystic adenoma (SMA) is a rare benign neoplasm. It accounts for 1-2% of all exocrine pancreatic tumors. It is thought to arise from the ductal epithelial cells. It is usually located in the body and the tail of the pancreas. It is important to identify SMAs and distinguish them from mucinous cystic neoplasm, which can be premalignant or malignant, and pseudocyst, which is a non neoplastic condition. We present one such rare case of SMA occurring in the head of the pancreas, an infrequent location requiring a Whippel's resection. PMID- 19679978 TI - A cytological and histomorphological case study of an uncommon breast carcinoma: Invasive papillary type. AB - Pure papillary carcinoma of the breast is a rare tumour affecting elderly postmenopausal women. We report one case in a relatively younger woman presenting with a clinically benign breast lump.The tumour showed extensive apocrine metaplasia. The ease with which abundant material with highly cellular papillary clumps is obtained on fine needle aspirate should be an important consideration favouring papillary carcinoma. The quality and quantity of stroma in papillae rather than the presence or absence of stromal support should also be a guiding criteria for excluding benign papillary lesions. PMID- 19679979 TI - Extradigital glomus tumor as a cause of chronic perianal pain. AB - Glomus tumor is a rare perivascular benign tumor arising from the Sucquet-Hoyer canal of the normal glomus body, most commonly in the digital areas. We report a serving soldier with such a tumor in an atypical site, the perianal region, presenting with episodic shooting pain. Total surgical excision was performed. Histopathology revealed a well-circumscribed tumor composed of clusters of monotonous polygonal cells surrounding capillary-sized blood vessels. Tumor cells also showed immunopositivity for smooth muscle antigen and vimentin. Following excision, the patient was completely relieved of pain and there was no recurrence on follow-up for 6 months. PMID- 19679980 TI - Pure choriocarcinoma of ovary diagnosed by fine needle aspiration cytology. AB - Pure ovarian choriocarcinoma is extremely rare and can develop as a germ cell tumor or as a metastasis from uterine or tubal gestational choriocarcinoma or rarely from an ovarian pregnancy. The cytomorphologic findings have been reported previously in different sites. However, this is the first case of pure ovarian choriocarcinoma diagnosed on cytology to the best of our knowledge. The distinction between a gestational and nongestational choriocarcinoma is difficult. A 19-year-old female patient presented with an irregular per-vaginal bleeding and a mass in lower abdomen. Fine needle aspiration cytology smears of the mass were hypocellular and showed large, multinucleated giant cells and malignant mononucleated cells. Background was hemorrhagic. Serum beta hCG level was 3,80,000 mIU/ml. A diagnosis of choriocarcinoma was offered which was later confirmed by histopathology. The diagnosis of choriocarcinoma on fine needle aspiration cytology is based on the presence of large, multinucleated giant cells and malignant mononucleated cells. A high index of suspicion should be maintained and estimation of serum beta hCG plays a key role in supporting the diagnosis. PMID- 19679981 TI - Hypergranular precursor B-cell acute lymphoblastic leukemia in a 16-year-old boy. AB - Presence of cytoplasmic granules in the blasts is a well known feature of myeloid leukemia. ALL presenting with the numerous cytoplasmic granules in blasts is a rarity and may be misdiagnosed as acute myeloid leukemia. We describe a rare case of hypergranular precursor B-cell acute lymphoblastic leukemia (ALL) in an adolescent male expressing CD10, CD19, CytoCD22, CD34, as well as CD13 and CD117. The blasts were cytochemically negative for myeloperoxidase (MPO), and acid phosphatase (ACP) but were positive for non-specific esterase (NSE). In centers where immunophenotypic panel is usually decided on the basis of morphology with limited antibodies may result in an erroneous typing of such rare diseases. Hence it is important to be aware of this rare entity and to confirm the lineage of acute leukemia by using a comprehensive panel of antibodies for immunophenotypic analysis. PMID- 19679982 TI - Congenital sideroblastic anemia: a report of two cases. AB - Sideroblastic anemia, comprising of acquired and congenital forms, is a heterogeneous group of disorders characterized by the presence of ring sideroblasts in the bone marrow. Congenital sideroblastic anemia is a rare condition which is mostly X-linked, caused by mutations of delta-aminolevulinic acid synthase 2. We describe two cases of congenital sideroblastic anemia, one of them indicating an autosomal recessive inheritance, with their clinico hematological profile. It is important to recognize this entity early in life as a significant percentage of cases respond to pyridoxine thus avoiding any long term complications. PMID- 19679983 TI - Iatrogenic Aspergillus infection of the central nervous system in a pregnant woman. AB - A healthy postnatal woman succumbed to fulminant iatrogenic Aspergillus infection of the central nervous system, following accidental inoculation into the subarachnoid space at spinal anesthesia, during an outbreak of Aspergillus meningitis in Sri Lanka. Autopsy revealed extensive Aspergillus meningitis and culture confirmed Aspergillus fumigatus. The thalamic parenchyma in the brain was invaded by fungal hyphae producing necrotizing angitis with thrombosis, thalamic infarcts and fungal abscesses. The directional growth of fungal hyphae from the extra-luminal side of blood vessels towards the lumen favored extension from the brain parenchyma over hematogenous spread. The spinal parenchyma was resistant to fungal invasion in spite of the heavy growth within the spinal meninges and initial inoculation at spinal level. Modulation of the immune response in pregnancy with depression of selective aspects of cell-mediated immunity probably contributed to rapid spread within the subarachnoid space, to involve the brain parenchyma leading to clinical deterioration and death. PMID- 19679984 TI - Urinary tract infection due to Enterobacter sakazakii. AB - Enterobacter sakazakii is a rare but important cause of necrotizing enterocolitis, bloodstream infection and central nervous system infections in humans, with mortality rates of 40-80%. It has not been reported to cause urinary tract infection. We report a case of urinary tract infection due to E. sakazakii in a 63-year-old lady with chronic renal failure. PMID- 19679985 TI - Primary spinal extradural hydatid cyst causing paraplegia. AB - Spinal hydatid disease is an uncommon cause of spinal cord compression and it constitutes 1% of all cases of hydatid disease. The authors present a case of a 21-year-old female presenting with rapid onset paraplegia caused by pathologically confirmed by extradural spinal hydatid cyst. Patient had marked improvement following surgical intervention. The case is discussed and the relevant literature is briefly reviewed. PMID- 19679986 TI - Salpingitis isthmica nodosa. PMID- 19679987 TI - Congenital epulis of the newborn. PMID- 19679988 TI - Pulmonary actinomycosis masquerading as tuberculosis. PMID- 19679989 TI - Renal and perinephric abscess due to Staphylococcus aureus. PMID- 19679990 TI - Gingival tuberculosis. PMID- 19679991 TI - Russian pathology and scientific misconduct. PMID- 19679992 TI - Calcified amorphous tumor of the heart. PMID- 19679993 TI - Paraphenylenediamine poisoning: laboratory medicine perspective. PMID- 19679994 TI - Metastatic hepatocellular carcinoma presenting as a gingival mass. PMID- 19679995 TI - Primary leiomyosarcoma of the penis. PMID- 19679996 TI - Idiopathic splenic arteriolar hyalinosis. PMID- 19679997 TI - Structuring data in anatomic pathology reports. PMID- 19679998 TI - Hb H disease: an under diagnosed entity in Indian setup. PMID- 19679999 TI - Increased emperipolesis in megakaryocytes in a case of idiopathic thrombocytopenic purpura. PMID- 19680000 TI - An unusual soft tissue plasmacytoma with massive intracellular deposition of needle-shaped crystals. PMID- 19680001 TI - Subcutaneous phaeohypomycosis. PMID- 19680002 TI - Beta-lactamase producing Acinetobacter species in hospitalized patients. PMID- 19680003 TI - Seroprevalence of Brucella agglutinins: a pilot study. PMID- 19680004 TI - Invasive cryptococcus infections in immunocompetent individuals may suggest defects in CD40/CD40L signaling pathway. PMID- 19680005 TI - Gynecomastia with unusual histology. PMID- 19680006 TI - Osteopontin and its clinical significance. PMID- 19680008 TI - A simulation study of radon and thoron discrimination problem in case-control studies. AB - In most countries, radon is the dominant contributor among natural radiation sources to the radiation exposure dose of the general population. Numerous case control studies of residential radon and lung cancer have been conducted using passive radon (Rn-222) detectors. These studies showed that radon may increase lung cancer risk, but most of them did not show a significant risk. Recently it was shown that the readings of passive radon detectors that do not employ thoron (Rn-220) discrimination techniques are affected by thoron. Therefore, we conducted a simulation study to evaluate the possible effect of thoron interference on the estimation of radon-related lung cancer risk. Various assumptions were made based on the number of cases, matching ratio, baseline risk, true radon-related risk, distribution of radon and thoron concentrations, correlation between radon and thoron, and radon detectors. The results suggested that in certain circumstances thoron interference in radon measurements resulted in an approximately 90% downward bias. In addition, the magnitude of the bias increased as the geometric mean and geometric standard error of radon concentration decreased and those of thoron increased. In order to resolve this problem, it is necessary to use passive radon detectors with thoron discrimination techniques in epidemiological studies. PMID- 19680007 TI - A case of abrupt onset autoimmune type 1 diabetes mimicking fulminant type 1 diabetes. AB - A 63-year-old male was admitted to our hospital with diabetic ketoacidosis. He had flu-like symptoms 10 days before admission and developed thirst, polyuria and anorexia with 9 kg of body weight loss in a week. Plasma glucose level on admission was 983 mg/dL and HbA1c was 7.5%. Despite high levels of serum pancreatic enzymes, lack of severe abdominal pain and no morphological change of pancreas in the abdominal CT scan eliminated the complication of classical acute pancreatitis. These findings suggested the diagnosis of fulminant type 1 diabetes. However, urinary and plasma C-peptide levels showed that insulin secretion was not completely depleted at onset. Furthermore, an examination of islet-related antibodies revealed the presence of high titer anti-GAD antibody. His HLA typing showed that DRB1*0901-DQB1*0303 and A24 were present. He has been doing well with continuation of insulin therapy. Over two years after onset, his plasma C-peptide level was gradually lowered, and anti-GAD antibody was still positive. Taken together, this is a rare case of abrupt onset autoimmune type 1 diabetes with transient but apparent exocrine pancreatic impairment at onset. Similar cases should be accumulated to clarify pathophysiological similarities and/or differences between fulminant type 1 diabetes and abrupt onset autoimmune type 1 diabetes. PMID- 19680009 TI - Low-dose radiation attenuates chemical mutagenesis in vivo. AB - The biological effects of low-dose radiation are not only of social concern but also of scientific interest. The radioadaptive response, which is defined as an increased radioresistance by prior exposure to low-dose radiation, has been extensively studied both in vitro and in vivo. Here we briefly review the radioadaptive response with respect to mutagenesis, survival rate, and carcinogenesis in vivo, and introduce our recent findings of cross adaptation in mouse thymic cells, that is, the suppressive effect of repeated low-dose radiation on mutation induction by the alkylating agent N-ethyl-N-nitrosourea. PMID- 19680010 TI - An approach to estimate radioadaptation from DSB repair efficiency. AB - In this review, we would like to introduce a unique approach for the estimation of radioadaptation. Recently, we proposed a new methodology for evaluating the repair efficiency of DNA double-strand breaks (DSB) using a model system. The model system can trace the fate of a single DSB, which is introduced within intron 4 of the TK gene on chromosome 17 in human lymphoblastoid TK6 cells by the expression of restriction enzyme I-SceI. This methodology was first applied to examine whether repair of the DSB (at the I-SceI site) can be influenced by low dose, low-dose rate gamma-ray irradiation. We found that such low-dose IR exposure could enhance the activity of DSB repair through homologous recombination (HR). HR activity was also enhanced due to the pre-IR irradiation under the established conditions for radioadaptation (50 mGy X-ray-6 h-I-SceI treatment). Therefore, radioadaptation might account for the reduced frequency of homozygous loss of heterozygosity (LOH) events observed in our previous experiment (50 mGy X-ray-6 h-2 Gy X-ray). We suggest that the present evaluation of DSB repair using this I-SceI system, may contribute to our overall understanding of radioadaptation. PMID- 19680012 TI - Live imaging of radiation-induced apoptosis by yolk injection of Acridine Orange in the developing optic tectum of medaka. AB - To observe the sequential radiation-induced apoptosis in a living embryo, we injected Acridine Orange (AO) solution into the yolk of embryo and visualized radiation-induced apoptosis in developing optic tectum (OT). Medaka embryos at stage 28, when neural cells proliferate rapidly in the OT, were irradiated with 5 Gy X-rays which is a non-lethal dose for irradiated embryos at hatching. The irradiated embryos hatched normally without morphological abnormalities in their brains, even though a large number of apoptotic cells were induced transiently in OT. By yolk injection, apoptotic cells in OT were distinguished as AO-positive small nuclei at 3 h after irradiation. At 8-10 h after irradiation, AO-positive rosette-shaped clusters were obviously distinguished in marginal tectal regions of OT where cells are proliferating intensely. The AO-positive clusters became bigger and more obvious, but the number did not increase up to 24 h after irradiation and completely disappeared up to 49 h after irradiation. This characteristic appearance of the AO-positive nuclei/clusters is in good agreement with our previous results, based on the examination of fixed specimens stained with AO by injection into the peri-vitelline space, suggesting that the AO-yolk injection method is highly reliable for detecting apoptotic cells in living embryos. The live imaging of apoptotic cells in developing Medaka embryos by AO yolk injection method is expected to reveal more of the details of the dynamics of apoptotic responses in the irradiated brain and other tissues. PMID- 19680011 TI - Radiation-quality dependent cellular response in mutation induction in normal human cells. AB - We studied cellular responses in normal human fibroblasts induced with low-dose (rate) or low-fluence irradiations of different radiation types, such as gamma rays, neutrons and high linear energy transfer (LET) heavy ions. The cells were pretreated with low-dose (rate) or low-fluence irradiations (approximately 1 mGy/7-8 h) of 137Cs gamma rays, 241Am-Be neutrons, helium, carbon and iron ions before irradiations with an X-ray challenging dose (1.5 Gy). Helium (LET = 2.3 keV/microm), carbon (LET = 13.3 keV/microm) and iron (LET = 200 keV/microm) ions were produced by the Heavy Ion Medical Accelerator in Chiba (HIMAC), Japan. No difference in cell-killing effect, measured by a colony forming assay, was observed among the pretreatment with different radiation types. In mutation induction, which was detected in the hypoxanthine-guanine phosphoribosyltransferase (hprt) locus to measure 6-thioguanine resistant clones, there was no difference in mutation frequency induced by the X-ray challenging dose between unpretreated and gamma-ray pretreated cells. In the case of the pretreatment of heavy ions, X-ray-induced mutation was around 1.8 times higher in helium-ion pretreated and 4.0 times higher in carbon-ion pretreated cells than in unpretreated cells (X-ray challenging dose alone). However, the mutation frequency in cells pretreated with iron ions was the same level as either unpretreated or gamma-ray pretreated cells. In contrast, it was reduced at 0.15 times in cells pretreated with neutrons when compared to unpretreated cells. The results show that cellular responses caused by the influence of hprt mutation induced in cells pretreated with low-dose-rate or low-fluence irradiations of different radiation types were radiation-quality dependent manner. PMID- 19680013 TI - Fifty years of gastroenterology at the University of Alabama at Birmingham: a festschrift for Dr. Basil I. Hirschowitz. PMID- 19680014 TI - Nonsteroidal antiinflammatory drugs and cyclooxygenase inhibition in the gastrointestinal tract: a trip from peptic ulcer to colon cancer. AB - Aspirin was commercialized more than a 100 years ago. Today, this compound is still widely prescribed, and new mechanisms of action and indications are being tested. Inhibition of cyclooxygenase (COX)-1 and COX-2 by aspirin or its related compounds, nonsteroidal antiinflammatory drugs (NSAIDs), has been associated with both adverse and beneficial effects in the gastrointestinal (GI) tract. Inhibition of COX-1 has been linked to GI adverse effects. Adverse effects of NSAIDs and aspirin in the upper GI tract include esophagitis, peptic ulcer, peptic ulcer complications, and death. Effective preventive therapies are available that have been associated with a progressive decline in the rate of hospitalization due to upper GI complications. NSAIDs and aspirin can also damage the small bowel and the colon. NSAID enteropathy is frequent and in most cases subclinical (increased mucosal permeability, inflammation, erosion, ulcer). However, more serious clinical outcomes such as anemia, bleeding, perforation, obstruction, diverticulitis, and deaths have also been described. Prevention therapy of NSAID damage to the lower GI tract is not well defined. Inhibition of COX-2 by NSAIDs, coxibs, or aspirin seems to provide beneficial effects to the GI tract. Observational studies show that these compounds reduce the risk of both upper and lower GI cancers. Randomized controlled trials have shown that aspirin and coxibs reduce the recurrence rate of colonic polyps, and long-term cohort studies have shown that aspirin reduces the risk of colon cancer time and dose dependently. New studies will have to define the appropriate population that may benefit with these therapies. PMID- 19680015 TI - Cancer stem cells: the other face of Janus. AB - Advances in the field of stem cell biology have provided renewed hopes that stem cells can be used to treat a wide range of genetic diseases and traumatic injuries. However, advances in the field of cancer cell biology have led to formulation of the cancer stem cell hypothesis, which posits that cancers arise from mutant stem cells. Further, this hypothesis proposes that these stem cells account for cancer recurrence, metastasis, and resistance to conventional treatments. Thus, although normal stem cells represent potential effective solutions to numerous clinical problems, when mutated, they may also represent the cause of many human malignancies. PMID- 19680016 TI - Breast abscesses in nonlactating women with diabetes: clinical features and outcome. AB - BACKGROUND: The association between diabetes and breast abscess in nonlactating women has not been previously reported in the literature. METHODS: Retrospective analysis of all cases of breast abscess in nonlactating women in a community teaching hospital from 2000 to 2006. We analyzed their clinical characteristics, prevalence of diabetes mellitus (DM), management, and clinical outcome. RESULTS: We identified 116 breast abscesses in 98 nonlactating women [age 48 +/- 14, (mean +/- SD), 89% African Americans]. At presentation, 63 patients (64%) had a known history of DM (duration: 8.5 +/- 5 years) and 8 patients (8%) had newly diagnosed DM. Among the remaining 27 women, 7 (26%) developed DM within 5 years of follow up. The odds ratio of having diabetes in nonlactating women with breast abscess relative to those without breast abscess was estimated as 14.24 (95% confidence interval, 6.72-30.17). Most patients (89%) had a single abscess. Patients with DM had increased length of hospital stay (P < 0.01) and a more severe clinical course during follow-up. Most patients (70%) were treated with incision and drainage and antibiotics. Glycemic control was suboptimal with only 46% of subjects receiving insulin therapy during the hospital stay. CONCLUSIONS: Our study indicates a strong association between DM and breast abscess in nonlactating women. The high prevalence of DM (72%) and their more severe clinical course suggest that breast abscesses in nonlactating women should be considered among the "typical" infections associated with DM. Increased awareness and intensified glycemic control might improve clinical outcome in nonlactating women with breast abscesses. PMID- 19680017 TI - Predictors for outcome and treatment delay in patients with tuberculous meningitis. AB - BACKGROUND: Tuberculous meningitis (TBM) is a challenge for clinicians because of the difficulty in making an early diagnosis and the severe consequences of delaying treatment. The objective of this study was to assess predictors of outcome and to evaluate factors critical to treatment delay of TBM. METHODS: One hundred and five adult patients with TBM, between 1997 and 2006, were retrospectively studied. Treatment delay was defined as progression of stage and physician delay between the initial presentation and the start of antituberculosis therapy. Factors contributing to the outcome, progression of stage, and prolonged physician delay were evaluated using univariate and multivariate analyses. RESULTS: Fifty patients (47.6%) experienced prolonged physician delay, and 38 (36.2%) had progression of stage. Thirty-four patients (32.4%) had an acute clinical course, and 76 (72.4%) received initial antibacterial therapy. Prolonged physician delay and progression of stage were important prognostic factors for poor outcome. Stage I at admission and prolonged physician delay were important factors contributing to progression of stage. An acute clinical course and an initial antibacterial therapy were important factors contributing to prolonged physician delay. CONCLUSIONS: Rapid diagnosis and early treatment before the occurrence of progression of stage are crucial for the outcome of TBM. TBM may present with an acute course, and when discrimination from bacterial meningitis is difficult, it is mandatory to start antituberculosis and antibacterial therapy simultaneously or lower the threshold for early antituberculosis therapy when persistent fever, deteriorated consciousness status, or progression of stage occurs during antibacterial therapy. PMID- 19680018 TI - Axial gouty arthropathy. AB - Gouty involvement of the spinal column is not as rare as generally perceived. Tophaceous gout involving the spinal column is a well-documented cause of myelopathy and frank cord compression. It takes several years of gout before bony destruction is radiologically apparent. If erosive or tophaceous gout is present, magnetic resonance imaging signal enhancement offers diagnostic guidance. Non tophaceous gout of the spine may also show signal enhancement consistent with inflammation. The sequelae of cord compression can be reversed with timely surgical intervention and maintenance of uric acid-lowering therapy; in some cases, medical therapy alone can reverse the findings of radiculopathy. Growing evidence suggests that the tangled web of hypertension, diabetes, and atherosclerotic disease are risk factors for gout and hyperuricemia and may, in fact, be the result of higher than physiologically tolerable levels of uric acid in humans. Here, 52 additional cases to the 73 collated by Hou et al (Surg Neurol. 2007;67:65-73), reinforce that gout is a major contender on the differential diagnosis of back-related presentations in patients at high risk for gout. The pervasiveness of cardiovascular disease and chronic back pain warrants a closer look into a possible occult contributor to the prevalence of chronic back pain: gout. PMID- 19680019 TI - Massive inguinal hernia. PMID- 19680020 TI - Landmark publication from The American Journal of the Medical Sciences: The significance of certain pulmonary lesions in relation to the etiology of influenza. PMID- 19680021 TI - Goodpasture's 1919 article on the etiology of influenza. PMID- 19680022 TI - Goodpasture's 1919 article on the etiology of influenza-the historical road to what we now call Goodpasture Syndrome. PMID- 19680023 TI - Transient ischemic attack in an elderly patient: what to expect? PMID- 19680024 TI - Masquerading myxoma. AB - Myxomas are the most common primary cardiac neoplasms. They are generally benign and most commonly arise from the left atrium. The clinical course of the left atrial myxoma is characterized by symptoms resulting from obstructive, embolic, or "constitutional" effects of the tumor (Goodwin, Lancet. 1963;1:464; Selzer et al, Am J Med. 1972;52:9; Nasser et al, Am Heart J. 1972;83:694). Obstructive symptoms are most common. Generally symptoms of the obstructive presentation are represented by dyspnea, pulmonary edema, or syncope. Embolic ischemic symptom manifestations are typically cerebral, although they could be peripheral (Greenwood, Am Heart J. 1972;83:694). Constitutional symptoms are seen in 50% of patients. Weight loss, low-grade fever, myalgia, arthralgia, and rash are typical nonspecific constitutional manifestations. Here, we describe an unusual clinical presentation of left atrial myxoma in a young boy, which initially was assessed as allergic dermatitis and later as vasculitis. PMID- 19680025 TI - Moxifloxacin-induced QT prolongation and torsades: an uncommon effect of a common drug. AB - Torsade de pointes (TdP) or "twisting of the points" represents polymorphic ventricular tachycardia in the setting of prolonged QT interval and is characterized by QRS complexes that change in morphology and amplitude. We report a rare case of TdP, associated with QT interval prolongation, caused by intravenous moxifloxacin given for pneumonia in a 71-year-old African American man. Electrocardiogram initially revealed QT interval prolongation that led to torsades de pointes. These changes reverted to normal when moxifloxacin was held. Although the risk for quinolone-associated TdP seems to be low, caution is still warranted when given someone with high risk of QT prolongation. PMID- 19680026 TI - Detection of left ventricular thrombus by coronary computed tomography angiography. AB - Left ventricular thrombus is a recognized complication of acute myocardial infarction. The following case report presents a rare case of left ventricular thrombus detection originally via coronary computed tomography angiography, followed by a brief review of imaging modalities that have been used for the detection of left ventricular thrombus in the past. PMID- 19680027 TI - Meningovascular syphilis with fatal vertebrobasilar occlusion. AB - We report the case of a young patient with meningovascular syphilis who suffered fatal vertebrobasilar occlusion despite thrombolytic treatment and endovascular interventions. A 35-year-old man without any known medical history presented with an acute ischemic stroke and was initially treated with intravenous tissue plasminogen activator. He was then transferred to the stroke center, where he underwent endovascular recanalization of his occluded vertebrobasilar system. Despite initial successful recanalization, he suffered recurrent vertebrobasilar occlusion, and a second endovascular treatment attempt was unsuccessful. He subsequently developed a pontine hemorrhage and acute hydrocephalus and died secondary to transtentorial herniation. Laboratory findings were suggestive of prior spirochetal infection, and autopsy revealed necrotizing vasculitis and extensive adventitial inflammation involving the basilar and vertebral arteries, supporting the diagnosis of meningovascular syphilis. PMID- 19680032 TI - Michela Baccini, winner of the Kenneth Rothman Epidemiology Prize, 2009. PMID- 19680033 TI - Epidemiology's contributions to a Nobel Prize recognition. PMID- 19680034 TI - Epidemiology and the elusive Nobel Prize. PMID- 19680035 TI - Economists and epidemiologists: variations on the Nobel pursuit. PMID- 19680036 TI - Opportunity knocks: the electronic (public health) medical record. PMID- 19680038 TI - A conversation with Maureen Henderson. Interview by Thomas Koepsell. PMID- 19680037 TI - Does air conditioning modify the health effects of exposure to outdoor air pollution? PMID- 19680039 TI - The remarkable Archie: origins of the Cochrane Collaboration. PMID- 19680040 TI - Childbearing and salivary gland cancer: a population-based nested case-control study. PMID- 19680041 TI - Saharan dust pollution: implications for the Sahel? [corrected]. PMID- 19680042 TI - Data sharing and scientific replication. PMID- 19680043 TI - Menstrual cycle characteristics and offspring sex ratio. PMID- 19680044 TI - Gestational diabetes and multiple sclerosis. PMID- 19680046 TI - Health care provider and consumer awareness, perceptions, and use of direct-to consumer personal genomic tests, United States, 2008. PMID- 19680047 TI - A critical discussion of the rates of progression and causes of optic nerve damage in glaucoma: International Glaucoma Think Tank II: July 25-26, 2008, Florence, Italy. AB - The International Glaucoma Think Tank II brought together glaucoma clinicians and researchers from all over the world to discuss current practices in glaucoma diagnosis and management, and the neurobiology of glaucoma. The meeting focused on several themes, including rates of deterioration in glaucoma patients, mechanisms of optic nerve damage, and implications for treatment. Issues such as how to measure and integrate progression information into clinical practice, screening protocols, or trials were discussed, as were promising new technologies and limitations of currently available measurement tools. Clinical applications for genetic testing were considered. Study of the neurobiology of glaucoma continues to inform our understanding of underlying degenerative processes, as well as to introduce possibilities for early detection or prevention. Many questions regarding glaucoma pathophysiology and best treatment practices remain unanswered, but with continued research and discussion, we will advance our understanding of this disease and ensure that patients receive optimal care. PMID- 19680049 TI - Effects of exercise on intraocular pressure and ocular blood flow: a review. AB - Glaucoma is a disease characterized by progressive optic neuropathy resulting in retinal ganglion cell death, which affects approximately 68 million people worldwide. Risk factors include intraocular pressure (IOP), genetics, race, age, and vascular factors. Exercise is known to affect IOP and systemic cardiovascular factors and, therefore, may affect glaucoma pathophysiology. This review discusses the results of articles relevant to glaucoma, IOP, ocular blood flow (OBF), and exercise. Isometric and dynamic exercises have been studied with respect to effects on IOP and OBF. Isometric exercise results in an acute decrease in IOP, which correlates with hypocapnia. Dynamic exercise results in a more pronounced but also short duration decrease in IOP. Physical fitness is associated with lower baseline IOP but diminished acute IOP-lowering response to exercise. Upon cessation of exercise, values return to pretrained levels within 1 month. In glaucoma patients, these IOP-lowering effects are greater than in healthy subjects. In healthy subjects, OBF is unchanged during exercise due to vascular autoregulation. This autoregulation fails at ocular perfusion pressures greater than 70% above baseline. In conclusion exercise in glaucoma patients results in acutely lowered IOP and lower baseline IOP. The effects of exercise on the prevention of glaucoma and glaucomatous progression remain unknown. The role of exercise in glaucoma management should be investigated. PMID- 19680048 TI - Lifestyle, nutrition, and glaucoma. AB - The only proven strategy to prevent primary open-angle glaucoma (POAG) is the use of ocular hypotensive therapy among people diagnosed with ocular hypertension. In this review, various modifiable lifestyle factors, such as exercise, diet, and cigarette smoking, that may influence intraocular pressure and that have been studied in relation to the risk of developing POAG are discussed. Epidemiologic studies on lifestyle factors are few, and the current evidence suggests that there are no environmental factors that are clearly associated with POAG; however, a few factors merit further study. This review also outlines future directions for research into the primary prevention of POAG. PMID- 19680051 TI - Level of vascular endothelial growth factor in aqueous humor and surgical results of ahmed glaucoma valve implantation in patients with neovascular glaucoma. AB - PURPOSE: To determine the associations between the levels of growth factors in aqueous humor and the surgical outcomes of Ahmed glaucoma valve implantation in patients with neovascular glaucoma (NVG). METHODS: From 19 NVG eyes of 19 patients, a sample of aqueous humor was taken just before Ahmed glaucoma valve implantation. Levels of vascular endothelial growth factor (VEGF), transforming growth factor (TGF)-beta1, and TGF-beta2 in aqueous humor aspirates were measured using a sandwich enzyme-linked immunosorbent assay method. When the postoperative intraocular pressures were >or=21 mm Hg with or without antiglaucoma medications at 2 successive visits or when any other surgical interventions were needed to lower intraocular pressure, that surgery was considered a failure. After patients were classified into success versus failure groups, the levels of the growth factors between the 2 groups was compared. The cumulative probability of success according to Kaplan-Meier analysis was also determined. RESULTS: Mean postoperative follow-up period was 40.9+/-19.6 months and cumulative probability of success was 43% at 57 months after surgery. Mean VEGF level in the failure group was higher than that of the success group (P=0.014). However, there was no statistical difference in the levels of TGF-beta1, TGF-beta2, and protein between 2 study groups (all P>0.05). CONCLUSIONS: The level of VEGF in aqueous humor was significantly higher in the failure group after the Ahmed glaucoma implantation compared with the success group. Our results imply that VEGF may play a role in determining surgical success after Ahmed valve implantation in patients with NVG. PMID- 19680050 TI - Repeatability of pattern electroretinogram measurements using a new paradigm optimized for glaucoma detection. AB - PURPOSE: To determine the within-trial and between-trial repeatability of pattern electroretinogram (PERG) measurements in healthy and patient eyes, using a new clinical instrument, the PERGLA. STUDY DESIGN: In all, 70 eyes of 35 healthy individuals (intraocular pressure <22 mm Hg, healthy optic disc by stereophotograph assessment, standard visual fields within normal limits) and 90 eyes of 45 clinic patients (ocular hypertensive, glaucomatous optic neuropathy by stereophotograph assessment and/or repeatable abnormal visual fields) enrolled in the University of California, San Diego Diagnostic Innovations in Glaucoma Study (DIGS) were evaluated. Average mean deviation of patient eyes on standard automated perimetry was -1.81 dB (SD=2.61). METHODS: The PERG was recorded using the PERGLA paradigm from both eyes simultaneously twice (ie, 2 trials) by a single operator with electrodes being removed and reattached between recordings. Repeatability of PERG amplitude (microV) and phase (pi rad) between 2 runs within a single trial (within-trial condition) was compared with repeatability between 2 trials (ie, after electrode replacement, between-trial condition) by calculating the coefficients of variability (CVs) and the intraclass correlation coefficients (ICCs) and displaying Bland-Altman plots. RESULTS: For healthy eyes, amplitude CVs (SD) were 11.5% (11.5) and 9.9% (0.79) for within-trial and between-trial conditions, respectively. ICCs were 0.91 and 0.85. Phase CVs were 1.3% (1.5) (within-trials) and 1.5% (1.4) (between-trials) and ICCs were 0.85 and 0.88. For patient eyes, amplitude CVs (SD) were 12.2% (10.1) and 11.2% (7.5) for within trial and between-trial conditions, respectively. ICCs were 0.92 and 0.89. Phase CVs were 2.2% (2.2) (within-trials) and 2.4% (2.2) (between-trials) and ICCs were 0.82 and 0.83. Bland-Altman plots indicated good agreement between the repeated recordings and were similar within-trials and between-trials for healthy and patient eyes. CONCLUSIONS: Repeatability of PERGLA recordings is good and is similar within-trials and between-trials for both healthy and patient eyes suggesting this technique is promising for monitoring change over time. PMID- 19680052 TI - Retinal nerve fiber layer thickness measurement by scanning laser polarimetry (GDxVCC) at conventional and modified diameter scans in normals, glaucoma suspects, and early glaucoma patients. AB - PURPOSE: To study the retinal nerve fiber layer (RNFL) at 3 different scan diameters using GDxVCC in healthy, primary open-angle glaucoma (POAG) suspects, and early POAG. METHOD: RNFL parameters were evaluated in 100 normal subjects, 50 POAG suspects, and 50 early POAG patients using GDxVCC at conventional small, medium, and large circle. TSNIT (temporal, superior, nasal, inferior, temporal) average, superior average, inferior average RNFL thickness, and nerve fiber indicator (NFI) were evaluated. RESULTS: The TSNIT average (microm) in normal subjects, POAG suspects, and POAG at 3 different circles was 54.17+/-4.6, 49.96+/ 6.5, 46.92+/-6.2 (small); 47.17+/-4.8, 43.79+/-5.7, 41.48+/-6.5 (medium); and 41.57+/-4.6, 39.77+/-5.9, 38.33+/-7.1 (large). All RNFL parameters at all circles showed significant difference between normal and early POAG eyes (P<0.001). Area under the curve was 0.911 for NFI at all circles. All parameters at small and medium circle, all except TSNIT average at large circle showed significant difference between normal and POAG suspects (P<0.001), area under curve being the highest (0.780 to 0.801 at 3 circles) for NFI. All parameters progressively decreased with increasing diameter with significant difference between small medium (P<0.001), medium-large (P<0.001), and small-large circle results (P<0.001) in all the groups for all parameters except NFI. CONCLUSIONS: Medium and large circles can also be used for RNFL evaluation in glaucoma. NFI is the best discriminating measure across all scan diameters. PMID- 19680053 TI - Evaluation of the position and function of aqueous drainage implants with magnetic resonance imaging. AB - PURPOSE: To evaluate position and function of antiglaucomatous Ahmed and Molteno shunts using magnetic resonance imaging with head and surface coils. METHODS: Eight patients (5 males) with shunt implants were included, 4 with Ahmed (FP-7) and 4 with Molteno (s1, single plated). All patients were operated at least 6 months before imaging. In 3 cases (2 with Molteno and 1 with Ahmed shunt), the intraocular pressure (IOP) was above 21 mm Hg, despite maximal medical treatment. The shunt endplate, tube and filtering blebs were identified in T1-weighted and T2-weighted images with both head and surface coils. Volumetric measurements of the orbits, eyeball, and filtering bleb and calculation of the endplate position along sagittal, transverse, and vertical axes were performed in T1-weighted and T2-weighted images using head coils. RESULTS: The shunt endplate was identified in T1-weighted and T2-weighted images (head coils) as a low intensity (dark) circumlinear band at the superotemporal aspect of the eyeball, surrounded by a pocket of water density, corresponding to the filtering bleb. The anterior position of the endplate, and smaller volume of the orbital cavity (less available orbital space) were associated with higher IOP. Filtering bleb volume was inversely correlated with IOP. In the unsuccessful cases, filtering bleb was absent. CONCLUSIONS: Magnetic resonance imaging provides insights into the mechanism of aqueous outflow and causes of failure of shunts. A lower orbital volume is associated with anterior position of the shunt endplate and poor shunt performance. PMID- 19680054 TI - Optic nerve head morphologic characteristics in chronic angle-closure glaucoma and normal-tension glaucoma. AB - PURPOSE: To determine whether the morphometric features of the optic disk in eyes with chronic angle-closure glaucoma (CACG) are different from those with normal tension glaucoma (NTG). PATIENTS AND METHODS: This is a retrospective, case matched study. Patients with CACG or NTG in an early or moderate stage were recruited. On the basis of age, sex, and visual field mean defect, patients with CACG were matched to those with NTG on a case-by-case basis. Heidelberg retina tomography was used to evaluate the optic disk, and parameters were compared between the 2 groups. RESULTS: Sixty-three patients were recruited for the study. Twenty cases (eyes) with CACG were matched to 20 cases (eyes) with NTG. The total, temporal/inferior, nasal/superior, and nasal/inferior rim area of the optic disk were significantly larger (P<0.05) in the CACG group. In patients with CACG rather than NTG, the cup volume in total and in each sector (except temporal) was significantly smaller (P=0.007 to 0.041). In patients with CACG rather than NTG, the mean cup depth and cup shape measure of the optic disk in total and in each sector (except temporal and temporal/inferior) were smaller (P=0.002 to 0.054). CONCLUSIONS: The morphometric features of the optic disk in eyes with CACG are different from those with NTG. PMID- 19680055 TI - Comparison of shape-based analysis of retinal nerve fiber layer data obtained From OCT and GDx-VCC. AB - PURPOSE: To directly compare in 1 population: (1) the performance of Optical Coherence Tomograph (OCT) and GDx-Variable Corneal Compensator (VCC) when using Wavelet-Fourier Analysis (WFA) and Fast-Fourier Analysis (FFA), (2) the performance of these shape-based and standard metrics, and (3) the shape of the retinal nerve fiber layer (RNFL) temporal, superior, nasal, inferior, temporal (TSNIT) curves obtained by the 2 different devices. METHODS: RNFL estimates were obtained from 136 eyes of 136 individuals (73 healthy and 63 mild glaucoma). WFA and FFA with and without asymmetry measures were performed on the TSNIT RNFL estimates to identify glaucoma from healthy eyes. Performance of WFA, FFA, and the standard metrics of OCT (Inferior Average) and GDX-VCC (Nerve Fiber Indicator) was evaluated by calculating receiver operating characteristic area. Measurements were obtained at a custom radius (33 to 41 pixels) for GDx-VCC to match the OCT radius (1.73 mm). RESULTS: WFA and FFA shape analysis significantly improved performance of both OCT (0.937) and GDx-VCC (0.913) compared with Inferior Average and Nerve Fiber Indicator (0.852 and 0.833, respectively). With either shape-based or standard metrics, OCT performance was slightly, but not significantly, better than GDx-VCC performance. Comparison of RNFL curves revealed that the GDx-VCC curves were more jagged and the peaks shifted more nasally when compared with the OCT RNFL curves. CONCLUSIONS: Performance of both OCT and GDx-VCC devices are improved by shape-based analysis methods. Classification performance was greater when using WFA for the OCT, and greater with FFA for the GDx-VCC. Significant differences between the machines exist in the measured TSNIT thicknesses, possibly because of GDx-VCC's measurements being affected by polarization magnitude varying with angle. PMID- 19680056 TI - Comparison of different modes in optical coherence tomography and ultrasound biomicroscopy in anterior chamber angle assessment. AB - PURPOSE: To compare anterior chamber angle width measurements by low and high resolution anterior segment optical coherence tomography (AS-OCT) and ultrasound biomicroscopy (UBM). PATIENTS AND METHODS: Thirty-six eyes of 32 patients with gonioscopically narrow angles from a university-based glaucoma clinic were recruited in the study. All subjects received AS-OCT {in "Anterior Segment Single" mode [low-resolution optical coherence tomography (LOCT)] and "High resolution Cornea" mode [high-resolution optical coherence tomography (HOCT)]} and UBM examination in 1 or both eyes. Angle opening distance (AOD) and angle recess area of temporal (t) and nasal (n) quadrants were assessed using the customized software on each type of image. Angle measurements from UBM, LOCT, and HOCT images were compared; intraobserver and interobserver measurement reproducibility for each type of image was also evaluated. RESULTS: Thirty-three eyes of 29 patients were available for analysis. No significant differences were found between LOCT and UBM in the measurement of t-AOD (P=0.350), n-AOD (P=0.106), and n-angle recess area (P=0.360). HOCT measurements were significantly larger than both LOCT and UBM for all parameters (all, P<0.05). HOCT images had better interobserver and intraobserver measurement reproducibility than UBM images. CONCLUSIONS: LOCT is similar to UBM for most of the studied angle measurements. HOCT tends to give larger measurements than both LOCT and UBM. AS-OCT measurements were more reproducible than those from UBM. PMID- 19680057 TI - Noncontact in vivo scanning laser microscopy of filtering blebs. AB - PURPOSE: To evaluate the ability of a noncontact, prototype scanning laser confocal microscope to image microstructural features of filtering blebs in vivo and to correlate these with the clinical features. METHODS: Thirty-one blebs of 24 patients underwent noncontact in vivo confocal microscopy using the Rostock Cornea Module of the Heidelberg Retina Tomograph II (50x noncontact Nikon lens, 1 2 mum resolution, field of view: 500x500 mum). Blebs were clinically classified as successful (low, diffuse, microcystic, and hypovascular) (n=22, mean intraocular pressure<20 mm Hg without hypotensive medications) or failed (flat, cicatrized, and nonfiltering) (n=9, mean intraocular pressure>20 mmHg with or without hypotensive medications). Multiple transverse images were taken over the scleral flap at 1 to 2 mm from the limbus. Images were evaluated by 2 observers masked to clinical appearance and function of the bleb. RESULTS: Transverse images of the successful blebs showed epithelial microcysts (50 to 100 microm) and small-sized to medium-sized (100 to 150 microm) subconjunctival cystic spaces with scattered loose collagen-like networks around the scleral flap. Failed blebs showed diffuse hyperreflective dense stroma, increased and tortuous vascularization, and no cystic spaces. The microscopic findings of all blebs were consistent with previous reports of contact in vivo confocal microscopy. CONCLUSIONS: Noncontact in vivo confocal microscopy of filtering blebs correlated with the clinical morphology. This noncontact approach provides information about bleb morphology without the risk of bleb microinjury, leak, or infection. Further study and optimization for imaging surface tissues are needed to improve our understanding of bleb maturation and wound healing. PMID- 19680058 TI - Replacement of Ahmed aqueous drainage devices in eyes with device-related complications. AB - PURPOSE: To examine the results of removal and simultaneous replacement of Ahmed aqueous drainage devices (ADDs), which require removal due to complications. METHODS: Noncomparative retrospective case series of 6 patients. RESULTS: For the period from January 1999 to December 2007, 325 Ahmed ADD insertions were performed in 272 patients. From this, we identified 6 patients (1.8%) who underwent removal of an ADD for device-associated complications. All had replacement of the Ahmed ADD in a different quadrant at the same surgery. The mean time interval from the original valve insertion was 31.2 months (median, 32.5; range, 3 to 67 mo). The indication for tube removal was chronic uveitis in 3 patients, plate exposure in 2 patients, and tube exposure in 1 patient. We examined the results at 12 months postsimultaneous removal and replacement of the ADD and at final follow-up (median, 25 mo; range, 13 to 52 mo). The preoperative complications resolved in all cases, with the inflammation settling postoperatively in the patients with preoperative uveitis and no patients developing tube or plate exposure at last follow-up. The mean preoperative intraocular pressure (IOP) was 16.0 mm Hg (median, 15.0; range, 9 to 29 mm Hg). At 12 months and final review, the mean IOP was 10.8 mm Hg (median, 10.0; range, 1 to 24 mm Hg) and 11.0 mm Hg (median, 10.0; range, 3 to 24 mm Hg), respectively. The mean number of glaucoma medications preoperatively was 2.8 (median, 3.5; range, 0 to 5). This was reduced to 1.7 (median, 1.5; range, 0 to 4) at 12 months and 2.2 (median, 1.5; range, 0 to 5) at final follow-up. Of the 6 cases in this series, 4 (66.6%) were considered to have adequate IOP control postoperatively. One patient developed postoperative hypotony, with an IOP of 4 mm Hg and reduced vision due to hypotony maculopathy. This patient declined further surgical intervention. One patient had a preoperative IOP of 29 mm Hg on 4 drops and acetazolamide 250 mg b.i.d. At 14 months postsurgery, IOP was 24 mm Hg on 4 drops and the patient underwent insertion of a second ADD. At last follow-up 14 months later, IOP was 10 mm Hg on 4 drops. There were no other significant intraoperative or postoperative complications in this series. CONCLUSIONS: In patients who require removal of Ahmed ADDs due to complications, removal of the offending ADD and replacement in another quadrant is effective in both resolving the complications and maintaining IOP control. PMID- 19680059 TI - Utility of bleb imaging with anterior segment optical coherence tomography in clinical decision-making after trabeculectomy. AB - AIM: To determine if imaging of blebs with anterior segment optical coherence tomography (ASOCT) affects clinical decision-making with regard to laser suture lysis (LSL) after trabeculectomy. METHOD: In this prospective observational case series, we included patients with poorly controlled intraocular pressure (IOP) after standardized trabeculectomy from May to November 2006. One observer assessed IOP, anterior chamber depth and bleb formation, and recorded a decision of whether or not to undertake LSL based on clinical grounds. A second observer masked to clinical data recorded a decision of whether or not to perform LSL based on ASOCT assessment of scleral flap position, presence of a sub-flap space, patency of the internal ostium, and bleb wall thickening. We compared the 2 observers' decisions to determine how ASOCT influenced decision-making. RESULTS: Seven eyes of 7 patients were included. On the basis of clinical examination, LSL was recommended in all 7 (100.0%) cases due to presence of elevated IOP, deep anterior chambers and poorly formed blebs. Using ASOCT, LSL was recommended in 5/7 (71.4%) cases with apposed scleral flaps, absent sub-flap spaces, and absent bleb wall thickening. In 2/7 (28.7%) cases, LSL was not recommended based on ASOCT findings of an elevated scleral flap, a patent sub-flap space, and bleb wall thickening. All 7 patients had good IOP control and formed blebs at a mean of 8.4+/-2.6 months after trabeculectomy, with a mean IOP of 14.3+/-3.2 mm Hg with no medications. CONCLUSIONS: This small study suggests that ASOCT imaging may affect decision-making with regard to LSL by providing information not apparent on clinical examination. PMID- 19680060 TI - Gemella species-associated late-onset endophthalmitis after trabeculectomy with adjunctive mitomycin C. AB - PURPOSE: To report 2 cases of delayed-onset, bleb-related endophthalmitis induced by Gemella species. METHODS: Interventional case report. RESULTS: Two patients developed bleb-related ocular infection attributable to Gemella haemolysans and Gemella morbillorum after trabeculectomy with adjunctive mitomycin C. Both patients had thin-walled cystic filtering blebs before ocular infection. One patient underwent urgent vitrectomy with intravitreal injection of ceftazidime and vancomycin. The other received an intraocular injection of vancomycin and ceftazidime. After surgical intervention, 2 cases were administered topical levofloxacin and sulfobenzylpenicillin, and intravenous cefpirome. The DNA from the microorganism was sequenced and identified as G. haemolysans and G. morbillorum. CONCLUSIONS: Gemella species should be added to the long list of organisms that may rarely cause bleb-associated endophthalmitis. PMID- 19680062 TI - Evidence-based practice. PMID- 19680063 TI - Effects of ankle-foot orthoses for children with hemiplegia on weight-bearing and functional ability. AB - PURPOSE: To compare the effects of a leaf spring ankle-foot orthosis (LAFO) and a hinged AFO with plantar flexion stop with that of shoe only on weight-bearing and function in children with hemiplegia. METHODS: The study used an experimental single-subject alternating treatment design with replication. Outcomes measured were weight-bearing symmetry, weight-bearing on the hemiplegic rear foot compared with that of the forefoot, functional ability, and child and parent preference. RESULTS: Both AFOs increased relative contact area of the hemiplegic foot. The LAFO increased relative force through the hemiplegic foot. There was increased pressure through the rear foot, time spent on the rear foot, and force through the forefoot. Rear foot contact area increased in the LAFO. Neither AFO affected function. Patient preference for AFO condition was inconsistent. CONCLUSION: Both AFOs increased weight-bearing through the hemiplegic foot and rear foot, indicative of potential benefit to growth. Neither the LAFO nor the hinged AFO with plantar flexion stop improved function of the children. PMID- 19680064 TI - Age expansion of the thirty-second walk test norms for children. AB - PURPOSE: The purposes of this study were to expand age ranges for a previously published normative database (n = 227) on the 30-second walk test, describe changes with age, explore contributions of subject characteristics, and verify previous data. METHODS: Children (n = 302; age, 5-17 years) from 4 urban schools were tested for distance walked in 30 seconds. Age, height, right lower extremity length, weight, sex, and race/ethnicity were recorded. RESULTS: Distance walked increased from 5 to 10 years of age, decreased slightly at age 11 years, followed by a more gradual decrease from 12 to 17 years. A significant difference in distance walked was found across ages. Right leg length, age, and weight explained 11.5% of the variance in walk distance. CONCLUSION: A percentile chart of the pooled data (previous and current, n = 529) should facilitate the use of the 30-second walk test when examining children for mobility limitations. PMID- 19680065 TI - The 30-second walk test (30sWT) norms for children. PMID- 19680066 TI - Parental and environmental factors associated with physical activity among children participating in an active video game. AB - PURPOSE: Parental and intervention-specific environmental supports were examined as potential reinforcers for physical activity and use of a video game, Dance Dance Revolution (DDR), among a cohort of 7- to 8-year-old children. METHODS: Sixty children were randomized to an intervention (n = 40) or a control (n = 20) group. Physical activity was measured with accelerometry and DDR logs. Parental support for their child's physical activity was assessed via a questionnaire. DDR specific environmental supports were captured on an environmental home screen and the DDR log. RESULTS: At baseline, the absence of other video games and parent DDR participation was associated with child participation in DDR. At follow-up, DDR participation of siblings and friends was associated with child participation in DDR. CONCLUSION: The primary findings of this study suggest that parental and peer participation in DDR may play a role in children's initial and sustained participation in DDR. PMID- 19680067 TI - Parents' perspectives on access to rehabilitation services for their children with special healthcare needs. AB - PURPOSE: This study examined parents' (caregivers') perspectives on problems related to access to therapy services for their children with special healthcare needs (CSHCN) as predicted by child, family, and health insurance characteristics. METHODS: Secondary data analysis was conducted using the Family Partners Project database. A subsample of 1027 parents of CSHCN who received rehabilitation services in the year before the study were the participants. RESULTS: Child, family, and insurance characteristics explained 19.1% of problems related to access to rehabilitation services with family characteristics being the strongest predictor (10.8%). Odds ratios are reported for these characteristics. CONCLUSION: Family financial hardship, the child's age, and managed care practices in health insurance plans may be the primary factors contributing to problems related to access to therapy services for CSHCN. Therapists may need a better understanding of family challenges with access to services to be more effective advocates. PMID- 19680068 TI - Development and reliability of an observational gait analysis tool for children with Down syndrome. AB - PURPOSE: To develop an observational gait analysis tool for children with Down syndrome and to examine its reliability. METHODS: Two physical therapists and 5 physical therapy students participated in this study. Sixty videos were examined to determine interrater reliability. Four months later, 15 of those 60 videos were reexamined twice (2 weeks apart) by the students for intrarater reliability. Intraclass correlation coefficients for interrater and intrarater reliability, percentage of agreement, and standard error of measurement were calculated. RESULTS: Interrater reliability was 0.663, intrarater reliability was 0.616 0.877, standard error of measurement was 1.89, and absolute agreement was found for 35.56% of the trials. CONCLUSION: Good reliability was found, but further study is needed to determine validity and clinical relevance. PMID- 19680069 TI - Listening to parents' concerns: three case examples of infants with developmental motor delays. AB - PURPOSE: The primary purpose of this clinical report was to compare parents' concerns about their infants' suspected motor delays with scores on the Harris Infant Neuromotor Test (HINT) and the Bayley-II Motor Scale. The second purpose was to determine whether there was agreement in categorizing the delays on the 2 tests. SUMMARY OF KEY POINTS: Three female infants with parent-identified concerns of motor delay were assessed on the HINT and Bayley-II Motor Scale. All showed significant delays, corroborating parental concerns. On the HINT, all scored >2 SD above the mean, being categorized as significantly delayed. On the Bayley-II Motor Scale, all demonstrated significantly delayed motor performance, ie, >2 SD below the mean. RECOMMENDATIONS FOR CLINICAL PRACTICE: Pediatric physical therapists should listen to parents' concerns about their infants with suspected motor delays and provide assessments to address these concerns. When appropriate, these infants should be referred for early intervention services. PMID- 19680070 TI - Clinical exercise testing in children and adolescents with cystic fibrosis. AB - PURPOSE: To review the most common field and laboratory exercise tests available for children and adolescents with cystic fibrosis (CF). METHODS: Relevant studies for this review were identified by electronic search of Medline and PubMed databases between the years 1958 and 2008. The bibliographies of all accessed publications were also searched. Key descriptors were cystic fibrosis, exercise testing, aerobic fitness, children, and adolescents. RESULTS: Five field tests were selected for presentation, including discussion of their strengths and weaknesses. Laboratory tests measuring aerobic and anaerobic responses to exercise in children with CF were also selected for presentation and discussed along with a summary of safety considerations for exercise testing of children with CF. CONCLUSION: Exercise testing is regarded an important prognostic tool in CF care. However, despite its beneficial effects, clinical exercise testing seems underused. Clinicians and their staff should encourage patients with CF to be physically active and recommend exercise testing annually. PMID- 19680071 TI - Factors affecting prescription and implementation of standing-frame programs by school-based physical therapists for children with impaired mobility. AB - PURPOSE: The purpose of this study was to investigate factors considered in the prescription and implementation of standing-frame programs by school-based physical therapists. METHODS: A 20-item survey was mailed to 500 members of the APTA Pediatric Section and School-Based Special Interest Group. Survey questions addressed standing-frame program prescription and perceived benefits. RESULTS: Response rate was 77.2%. A majority of respondents rated ambulatory status for the prescription of standing-frame programs and a child's specific needs in the selection of a specific standing frame as very important. Respondents identified multiple benefits with pressure relief rated very important most frequently. More than 50% of respondents indicated social and educational benefits are very important. A majority of respondents prescribed standing-frame programs for 30-45 minutes daily. CONCLUSIONS: Variation does exist, but the majority of school based physical therapists agree on several key factors in the prescription and implementation of standing-frame programs. PMID- 19680073 TI - Letter to the editor. Lower extremity neuromuscular electrical stimulation. PMID- 19680074 TI - What's new in Shock, September 2009? PMID- 19680075 TI - Thalidomide suppresses sclerosing encapsulating peritonitis in a rat experimental model. AB - Peritoneal dialysis is an alternative treatment of patients with end-stage renal disease. Sclerosing encapsulating peritonitis is a life-threatening complication of continuous ambulatory peritoneal dialysis. The aim of the present study was to evaluate the effect of thalidomide, which is used for the treatment of various inflammatory and autoimmune diseases, on the development of sclerosing encapsulating peritonitis induced by chlorhexidine gluconate (CG). A peritoneal fibrosis model was established using rats treated intraperitoneally with injections of CG. Thalidomide was administered orally at a dose of 100 mg/kg three times per week. When compared with CG-treated rats, thalidomide (100 mg/kg orally)-treated mice subjected to CG-induced peritoneal fibrosis experienced a significantly lower rate in the extent and severity of histological signs of peritoneal injury. Thalidomide also caused a substantial reduction of 1) the rise in myeloperoxidase activity (mucosa); 2) the expression in the tissue of TNF alpha, IL-1beta, transforming growth factor-beta, and vascular endothelial growth factor; 3) the increase in staining (immunohistochemistry) for nitrotyrosine and for poly(ADP ribose), as well as 4) the nuclear factor-kappaB activation caused by CG in the peritoneum. Thus, thalidomide treatment reduces the degree of peritoneal fibrosis caused by CG. We propose that this evidence may help clarify the potential therapeutic actions of thalidomide in patients with peritoneal fibrosis. PMID- 19680077 TI - A large-bolus injection, but not a continuous infusion of sodium selenite improves outcome in peritonitis. Shock 32: 140-146, 2009. PMID- 19680078 TI - Creating new solutions to the simulation puzzle. PMID- 19680079 TI - Alternative educational models for interdisciplinary student teams. AB - BACKGROUND: Few studies compare instructor-modeled learning with modified debriefing to self-directed learning with facilitated debriefing during team simulated clinical scenarios. OBJECTIVE: : To determine whether self-directed learning with facilitated debriefing during team-simulated clinical scenarios (group A) has better outcomes compared with instructor-modeled learning with modified debriefing (group B). METHODS: This study used a convenience sample of students. The four tools used assessed pre/post knowledge, satisfaction, technical, and team behaviors. Thirteen interdisciplinary student teams participated: seven in group A and six in group B. Student teams consisted of one nurse practitioner student, one registered nurse student, one social work student, and one respiratory therapy student. The Knowledge Assessment Tool was analyzed by student profession. RESULTS: There were no statistically significant differences within each student profession group on the Knowledge Assessment Tool. Group B was significantly more satisfied than group A (P = 0.01). Group B registered nurses and social worker students were significantly more satisfied than group A (30.0 +/- 0.50 vs. 26.2 +/- 3.0, P = 0.03 and 28.0 +/- 2.0 vs. 24.0 +/- 3.3, P = 0.04, respectively). Group B had significantly better scores than group A on 8 of the 11 components of the Technical Evaluation Tool; group B intervened more quickly. Group B had significantly higher scores on 8 of 10 components of the Behavioral Assessment Tool and overall team scores. CONCLUSION: The data suggest that instructor-modeling learning with modified debriefing is more effective than self-directed learning with facilitated debriefing during team-simulated clinical scenarios. PMID- 19680080 TI - Action research, simulation, team communication, and bringing the tacit into voice society for simulation in healthcare. AB - BACKGROUND: In healthcare, professionals usually function in a time-constrained paradigm because of the nature of care delivery functions and the acute patient populations usually in need of emergent and urgent care. This leaves little, if no time for team reflection, or team processing as a collaborative action. Simulation can be used to create a safe space as a structure for recognition and innovation to continue to develop a culture of safety for healthcare delivery and patient care. METHODS: To create and develop a safe space, three qualitative modified action research institutional review board-approved studies were developed using simulation to explore team communication as an unfolding in the acute care environment of the operating room. An action heuristic was used for data collection by capturing the participants' narratives in the form of collaborative recall and reflection to standardize task, process, and language. RESULTS: During the qualitative simulations, the team participants identified and changed multiple tasks, process, and language items. The simulations contributed to positive changes for task and efficiencies, team interactions, and overall functionality of the team. CONCLUSION: The studies demonstrated that simulation can be used in healthcare to define safe spaces to practice, reflect, and develop collaborative relationships, which contribute to the realization of a culture of safety. PMID- 19680081 TI - Measures of stress and learning seem to be equally affected among all roles in a simulation scenario. AB - INTRODUCTION: Simulation scenarios provide a realistic, stressful environment in which participants can gain new clinical knowledge. It is unclear whether the role a participant plays in a scenario affects these goals. We measured heart rate, self-perceived stress and learning value, and objective written test results of participants in two simulation scenarios. Our hypothesis was that measurements of participants' stress and learning would be similar among all team members in our simulated scenarios. METHODS: Residents and medical students were prospectively randomized to take part in two difficult airway scenarios in the roles of team leader, procedure chief, or team member. Heart rate was recorded using pulse oximetry preprocedure and at the critical airway intervention. After debriefing, participants completed a data collection form that included visual analog scales for perceived stress and learning and objective questions related to scenario teaching points. RESULTS: We obtained 53 measurements during a single day. Heart rates increased during the airway intervention (median 4 beats per minute, P = 0.04). Self-reported learning values increased with self-reported stress level (rs = 0.373, P = 0.01); however, no correlation was found between a participant's role in the scenario and heart rate, test score, stress level, or perceived learning benefit. CONCLUSIONS: In our limited scenarios, measurements of stress and learning did not differ by role. Our results support the concept that all participants may benefit from simulation scenarios. PMID- 19680082 TI - Simulation training improves medical students' learning experiences when performing real vaginal deliveries. AB - OBJECTIVE: To determine the relationship between simulation training for vaginal delivery maneuvers and subsequent participation in live deliveries during the clinical rotation and to assess medical students' performance and confidence in vaginal delivery maneuvers with and without simulation training. METHODS: Medical students were randomized to receive or not to receive simulation training for vaginal delivery maneuvers on a mannequin simulator at the start of a 6-week clerkship. Both groups received traditional didactic and clinical teaching. One researcher, blinded to randomization, scored student competence of delivery maneuvers and overall delivery performance on simulator. Delivery performance was scored (1-5, with 5 being the highest) at weeks 1 and 5 of the clerkship. Students were surveyed to assess self-confidence in the ability to perform delivery maneuvers at weeks 1 and 5, and participation in live deliveries was evaluated using student obstetric patient logs. RESULTS: Thirty-three students were randomized, 18 to simulation training [simulation group (SIM)] and 15 to no simulation training [control group (CON)]. Clerkship logs demonstrated that SIM students participated in more deliveries than CON students (9.8 +/- 3.7 versus 6.2 +/- 2.8, P < 0.005). SIM reported increased confidence in ability to perform a vaginal delivery, when compared with CON at the end of the clerkship (3.81 +/- 0.83 versus 3.00 +/- 1.0, respectively, P < 0.05). The overall delivery performance score was significantly higher in SIM, when compared with CON at week 1 (3.94 +/- 0.94 versus 2.07 +/- 1.22, respectively, P < 0.001) and week 5 (4.88 +/- 0.33 versus 4.31 +/- 0.63, P < 0.001) in the simulated environment. CONCLUSIONS: Students who receive simulation training participate more actively in the clinical environment during the course of the clerkship. Student simulation training is beneficial to learn obstetric skills in a minimal risk environment, demonstrate competency with maneuvers, and translate this competence into increased clinical participation and confidence. PMID- 19680084 TI - Building team and technical competency for obstetric emergencies: the mobile obstetric emergencies simulator (MOES) system. AB - The infrequent and high-stakes nature of obstetric emergencies requires staff members to respond quickly and proficiently to a complex and high-stress situation, a situation they have likely had little opportunity to experience. This situation requires a systematic approach to preparing personnel to manage these situations. Therefore, this article seeks to contribute to the growing literature on training programs for obstetric emergencies by documenting the development and implementation of the Mobile Obstetric Emergencies Simulator (MOES) system. MOES is a comprehensive package of simulation technology, standardized curriculum, and instructional features that combines traditional classroom learning activities and simulation-based training on the actual labor and delivery (L&D) ward. Specifically, the MOES system leverages the TeamSTEPPS teamwork training being implemented throughout the US military healthcare system with opportunities to practice teamwork and technical skills using mannequin based patient simulation embedded within L&D units. The primary goals of this article are twofold. First, this article explicitly identifies the unique training needs for preparing staff for obstetric emergencies through a comprehensive review and synthesis of the literature. Second, this article documents the approach taken in MOES to meet these needs. PMID- 19680083 TI - Comparison of intubation success of video laryngoscopy versus direct laryngoscopy in the difficult airway using high-fidelity simulation. AB - INTRODUCTION: A number of devices, including video laryngoscopy, are used to aid in managing difficult airways. The goal of this study was to compare timing and success of video laryngoscopy to standard laryngoscopic intubation using a simulation mannequin in normal and difficult airway scenarios. METHODS: Residents and attending physicians of a PGY 2-4 emergency medicine residency program participated. A single, high-fidelity simulation mannequin was used. Each participant received an in-service on the video laryngoscope (GlideScope). Three airway settings were used: standard, decreased neck mobility, and tongue edema. Participants intubated with a Macintosh blade and video laryngoscope in each scenario, and graded the best view achieved using the Cormack-Lehane classification. Outcome measures included time to view the vocal cords, time to intubation, grading of view, and intubation success or failure. Institutional Review Board approval was obtained. RESULTS: Fifty-two participants were enrolled. Participants successfully intubated the mannequin faster using the Macintosh blade in both the normal and neck immobility settings (9.4 seconds faster, 95% CI 3.2-15.7, P = 0.004, 16.1 seconds faster, 95% CI 3.6-28.7, P = 0.01). In the tongue edema setting, however, video laryngoscopy provided a better grade view of the cords, a higher success rate of viewing the cords at time of intubation (50% vs. 12%), and a higher rate of successful intubations (83% vs. 23%). The GlideScope also significantly reduced the time needed to view the cords (89 seconds reduction, 95% CI 54.4-123.7, P < 0.0001) and intubate (131.3 seconds reduction, 95% CI 99.1-163.6, P< 0.0001) for the tongue edema setting. CONCLUSIONS: In the most difficult airway case, tongue edema, the video laryngoscope provided an enhanced view of the cords using less time, increased intubation success, and decreased the time to intubation. PMID- 19680085 TI - Modification of the Laerdal SimBaby to include an integrated umbilical cannulation task trainer. AB - BACKGROUND: Given the emphasis on early vascular access via the umbilical vein in neonatal resuscitation it is essential that participants in neonatal resuscitation simulation training be given the opportunity to practice both the placement and use of an emergency umbilical venous catheter. By integrating available parts from the Laerdal catalog, combined with a few other inexpensive components, into a Laerdal SimBaby we were able to create a single, integrated neonatal simulator that could be used to practice both the placement and use of an emergent umbilical vein catheter. METHODS: To integrate an umbilical cannulation task trainer into the Laerdal SimBaby we used a specially modified replaceable umbilical cord and reservoir from the Laerdal Neonatal Resuscitation Baby. To this reservoir we attached a flanged outlet drain and drainage tube which allows for the infusion of medications and fluids. The modified SimBaby with integrated umbilical cannulation task trainer was validated for both face and content by simulation participants and a panel of neonatal resuscitation experts. RESULTS: The umbilical cannulation task trainer integrated well into the SimBaby and in no way altered the function of the mannequin. The modified Laerdal SimBaby was thought to work well by both participants and experts. Simulation participants liked having the chance to practice emergency umbilical vein cannulation and thought that the simulated umbilical cord was an important component in their learning experience. The expert panel thought that the modified SimBaby could be used for emergency umbilical vein cannulation skills training and that the addition gave the modified SimBaby major advantage over other simulators they had used to teach newborn resuscitation. DISCUSSION: We have developed a modification to the Laerdal SimBaby involving the integration of a usable umbilical cannulation task trainer. The modification was easily accomplished using available parts from the Laerdal catalog and a few other inexpensive components. Given the emphasis on early vascular access via the umbilical vein and the complexities involved with the administration of medications and fluids via this route we believe that a usable umbilical cannulation task trainer is essential to neonatal resuscitation simulation training. When modified as described the Laerdal SimBaby can act as a high fidelity newborn simulator that allows participants to practice both the placement and use of an emergency umbilical vessel catheter. Given our positive experience we think others could apply the above modification to their own SimBaby. PMID- 19680086 TI - High spinal in an obstetric patient: a simulated emergency. PMID- 19680088 TI - Restoration of bone turnover rate after decompression surgery in patients with symptomatic lumbar spinal stenosis: preliminary report. AB - STUDY DESIGN: Prospective short-term longitudinal study. OBJECTIVE: To investigate changes in the bone turnover rate in patients with lumbar spinal stenosis (LSS) before and after decompression surgery. SUMMARY OF BACK GROUND DATA: Decompression surgery enables elderly patients with LSS to participate in daily activities and physical exercise by reducing or alleviating leg and back pain. However, there have been no studies to date regarding the effect of decompression surgery on bone metabolism in such patients. METHODS: Twenty-three patients with spinal stenosis who were scheduled to undergo decompression surgery were enrolled in our study. Ten patients were given oral bisphosphonates after the operation (B+ group), while the remaining 13 patients did not receive oral bisphosphonate (B- group). In both groups, walking distance without rest, the Oswestry Disability Index (ODI) scores, duration of symptoms, bone formation, and resorption markers, and bone mineral density were recorded before surgery. Three months after surgery, bone turnover markers, a single trial for walking distance without rest and ODI scores were measured for both groups. RESULTS: Three months after the operation, the bone resorption marker u-NTx was decreased significantly for both groups. Although there was a decrease in bALP, a bone formation maker, in both groups, the change in each group was not statistical significant. Distance in a single trial walk was increased and ODI scores were decreased significantly for both groups. CONCLUSION: This study suggests that decompression surgery has a beneficial effect on bone metabolism in patients with LSS who have walking intolerance and limited physical activity. PMID- 19680089 TI - Biomechanical evaluation of the vertebral jack tool and the inflatable bone tamp for reduction of osteoporotic spine fractures. AB - STUDY DESIGN: Controlled in vitro study. OBJECTIVE: To compare two kyphoplasty techniques in cadaveric fractured vertebrae: an experimental vertebral jack tool (VJT) and an inflatable bone tamp (IBT). SUMMARY OF BACKGROUND DATA: A previous biomechanical study showed restored strength and stiffness after height restoration in cadaveric-fractured osteoporotic vertebrae using a new device for reduction of osteoporotic vertebral fractures. METHODS: Anterior wedge fractures (AO type A1.2) were created in 8 (4 lumbar, 4 thoracic) vertebrae by displacement eccentric external forces. In all vertebrae the amount of height reduction was 35%. After compression, 4 vertebrae were restored in height using the VJT procedure. Four vertebrae were restored in height using the IBT procedure. Posttreatment strength and stiffness of the vertebrae were determined by a compression test identical to the pretreatment compression protocol. RESULTS.: In the VJT group the post-restoration strength was 81% +/- 13% of the original strength and in the IBT group it was 96% +/- 32%. The post-restoration stiffness in the VJT group was 61% +/- 42% of the original stiffness and in the IBT group 76% +/- 62% of the original stiffness. The vertebrae in the VJT group were restored to 101% +/- 2% of their original height whereas this was 104% +/- 14% in the IBT group. In this study, no cases of cement leakage were found. No cases of damaging of the end plates, new fractures or perforations were seen in both groups. The mean amount of cement inserted for the VJT group was 3.6 +/- 0.9 cm and for the IBT group 5.9 +/- 0.8 cm. CONCLUSION: Both kyphoplasty procedures were able to restore height in this in vitro study, while strength and stiffness were partially restored, with no significant differences. In this study on average significant less cement was used in the VJT procedure. No complications were noted in both groups. This new end plate-to-end plate laminar augmentation technique may be of clinical advantage. PMID- 19680090 TI - Cerebrospinal fluid concentrations of nitric oxide metabolites in spinal cord injury. AB - STUDY DESIGN: Multiple center study to evaluate cerebrospinal fluid (CSF) concentrations of nitric oxide metabolites [NOx] in relation to neurologic severity and prognosis in spinal cord injury (SCI). OBJECTIVE: To examine whether CSF [NOx] correlates with neurologic severity and recovery in SCI. SUMMARY OF BACKGROUND DATA: Inducible nitric oxide synthase is expressed in rat spinal cord immediately after SCI. Excessive nitric oxide production is cytotoxic, causing neuronal apoptosis with subsequent neurodysfunction in the spinal cord. We previously reported a significant correlation between initial [NOx] after incomplete cervical cord injury (CCI) and neurologic recovery at the final follow up in 25 cases. METHODS: Ninety-six cases (SCI group), including 76 patients with CCI and 20 patients with thoracic cord injury were examined. Mean follow-up period was 11 months. The control group comprised 40 cases (3 healthy volunteers and 37 patients with neither pain nor neurologic disorders). CSF [NOx] were measured using the Griess method. Severity of neurologic impairment was assessed using Frankel's classification and the American Spinal Injury Association motor score (ASIA MS). Degree of neurologic recovery was assessed using Frankel's classification and the ASIA motor recovery percentage. RESULTS: CSF [NOx] did not differ significantly among the control, CCI, and thoracic cord injury groups at the initial examination. In the CCI group, [NOx] in the Frankel A and B classes were significantly higher than [NOx] in the control group at 5 to 14 days, in the Frankel A and B classes at 0 to 4 days, and in the Frankel C and D classes at 5 to 14 days. Also, in the CCI group at 5 to 14 days, [NOx] correlated significantly with ASIA MS and motor recovery percentage. CONCLUSION: There was a significant correlation between CSF [NOx] at the pathologic early subacute stage (approximately 5-14 days) and neurologic severity and recovery in SCI. PMID- 19680091 TI - Dissecting the effects of spinal fusion and deformity magnitude on quality of life in patients with adolescent idiopathic scoliosis. AB - STUDY DESIGN: A retrospective review of scores from the Scoliosis Research Society outcomes instrument (SRS-24 questionnaire). OBJECTIVE: To quantify the isolated effects of spinal fusion and deformity magnitude on quality of life in patients with adolescent idiopathic scoliosis (AIS). SUMMARY OF BACKGROUND DATA: Significant improvements in 2-year postoperative SRS-24 questionnaire scores have been reported despite the loss of spinal motion due to instrumentation and arthrodesis. As deformity reduction may influence patient perception, it has been difficult to isolate the effect of spinal fusion on quality of life after scoliosis surgery. METHODS: SRS-24 scores were compared between 3 cohorts of AIS patients (preoperative, postoperative, and nonoperative) using an ANOVA (P < 0.05) to determine the isolated effects of spinal fusion and deformity magnitude. Preoperative SRS-24 scores were collected from a group of patients with preoperative major Cobb angles greater than 40 degrees (n = 194). Postoperative SRS-24 scores were collected from patients with preoperative major Cobb angles greater than 40 degrees and 2-year postoperative major Cobb angles between 20 degrees and 40 degrees (n = 196). Finally, SRS-24 scores were collected from a nonoperative group of patients with major Cobb angles between 20 degrees and 40 degrees (n = 112). RESULTS: Spinal fusion was found to have a negative isolated effect on the Activity domain (-0.3) and on the Total score (-0.2) (P = 0.001) of the SRS-24 questionnaire (score range: 1-5). A smaller deformity magnitude, on the other hand, was found to have a significantly positive isolated effect on all 4 preoperative domains (P < 0.001) and on the Total score (P < 0.001). The combined effect of surgery (spinal fusion and deformity correction) was found to be significantly positive for the Total score (P < 0.001) and for the domains of Pain, Self-Image, and Function (P < 0.001). CONCLUSION: Spinal fusion has an isolated negative effect on AIS patients' quality of life (Total score) mostly due to a decrease in scores of the Activity domain. The overall positive effect of surgery depends on the individual effects of spinal fusion (slight reduction in quality of life) and deformity reduction (modest improvement in quality of life). PMID- 19680092 TI - Comparison of the melatonin and calmodulin in paravertebral muscle and platelets of patients with or without adolescent idiopathic scoliosis. AB - STUDY DESIGN: Controlled clinical study. OBJECTIVE.: To compare muscle and platelet calmodulin and melatonin concentrations of scoliotic and nonscoliotic populations. SUMMARY OF BACKGROUND DATA: Melatonin and calmodulin are potential key molecules in scoliosis etiology. Calmodulin is not only a second messenger of melatonin but also has been shown to have effects on muscle contractility. There is a possibility that it may be of importance in the regulation of spinal alignment. Platelets have been defined as mini muscles calmodulin and melatonin levels of which may be the projections of muscle values. METHODS: Twenty patients undergoing posterior surgery for adolescent idiopathic scoliosis (AIS) and 9 thoracic-lumbar trauma patients undergoing posterior surgery constituted the population. Autologous bloods were collected and processed to obtain platelets. Paravertebral muscle tissue samples from both sides were obtained at T12-L1 level intraoperatively. Muscle and platelet samples were analyzed for the levels of melatonin by radio immuno assay and for calmodulin by enzyme-linked immunosorbent analysis. Groups, concave (left side for the control group) and convex side (right side for the control group), muscles and platelet median protein concentrations, and optic densitometry (OD) ratio values were compared. RESULTS: AIS group consisted of 2 male and 18 female patients. Mean age was 16.1 +/- 3.78 (11-29). Control group consisted of 5 male and 4 female patients. Mean age was 35 +/- 13.47 (16-55). Platelet Calmodulin OD/Supernatant's OD ratios and both convex and concave sides' muscle Calmodulin OD/Supernatants' OD ratios were not different between groups. On the other hand, convex side muscle calmodulin to total muscle calmodulin ratios were higher in AIS group compared with concave (P = 0.048); likewise, concave side calmodulin to total calmodulin ratios were lower in AIS group compared with control (P = 0.035). Convex side calmodulin to concave side calmodulin ratios were significantly different among groups (P = 0.048). Neither platelet melatonin to total protein ratios, nor convex or concave side muscle melatonin to total protein ratios, nor convex to concave side melatonin ratios were significantly different between groups. Convex or concave side calmodulin or melatonin values were not correlated with platelet values. CONCLUSION: AIS group had an asymmetric distribution of calmodulin in paraspinal muscle, higher at the convex side and lower at the concave. Neither platelet melatonin nor platelet calmodulin was found to be representative of the muscle protein values. PMID- 19680093 TI - Tubular microsurgery for lumbar discectomies and laminectomies in obese patients: operative results and outcome. AB - STUDY DESIGN: This is a retrospective single-center case study involving 115 obese and nonobese patients who underwent minimally invasive lumbar surgery between 2004 and 2007. OBJECTIVE: The purpose of this study was to evaluate the effect of minimally invasive spinal surgery in obese and nonobese patients by operative results and patient outcomes. SUMMARY OF BACKGROUND DATA: Spinal surgery in obese patients is associated with increased complications, blood loss, and operative times. The potential benefits of minimally invasive lumbar surgery in obese patients are discussed. METHODS: All patients underwent 1-level lumbar microdiscectomy or laminectomy using tubular retractors. Data were collected on patient demographics, comorbidities, smoking habits, operative results, and clinical outcomes, and compared for obese and nonobese patients. Operative results included operative times, blood loss, length of stay, and perioperative complications. Clinical outcomes were assessed by using pre- and postoperative visual analog scale and Macnab outcome criteria at most recent follow-up. RESULTS: In this study, 31% of 115 patients were classified as obese. Obese patients tended to undergo surgery at a younger age. Obesity, comorbidities, and age did not have an impact on patient outcome at a mean follow-up of 15.9 months. No significant differences were seen between obese versus nonobese patients in terms of incision lengths, operative time, blood loss, and complication rates. In obese patients, all parameters and operative results compared favorably to reported historical results of patients undergoing open lumbar surgery. Overall, favorable outcome was seen in 92% and 84% of obese and nonobese lumbar microdiscectomy patients, respectively, and in 75% of laminectomy patients. Postoperative visual analog scale did not show any significant difference. CONCLUSION: This is the first study comparing operative results from tubular microsurgery between obese and nonobese patients. No major differences were detected in outcome, operative and perioperative data including complication rates. With tubular microsurgery, obese patients experienced the same or equally beneficial outcome, compared to nonobese patients, while incision lengths, blood loss, operative times, and length of stay were less when compared to open procedures. Other comorbidities and age had no significant impact on perioperative complications and clinical outcome. PMID- 19680094 TI - Spinal subarachnoid hematoma with hyperextension lumbar fracture in diffuse idiopathic skeletal hyperostosis: a case report. AB - STUDY DESIGN: A case report of a spinal subarachnoid hematoma (SSAH) associated with hyperextension fractures complicating diffuse idiopathic skeletal hyperostosis (DISH). OBJECTIVE: To report the first case of a SSAH complicating spinal fracture in DISH. SUMMARY OF BACKGROUND DATA: A SSAH is very rare condition. And there had been several reports on spinal fractures in DISH through fused spinal segments, but no report related with SSAH associated with spinal fracture in DISH patient. METHODS: A 78-year-old female patient was admitted for pain in the back and lower limbs and paraparesis after being fall on her back. On simple radiographs, DISH with anterior cortical bone deficit and increased height were presented at the level of L1. MR and Myelography computed tomography images revealed an extension type of fracture with an irregular shaped subarachnoid hematoma within the dura. Differential diagnosis from infection or tumorous condition was required. RESULTS: Durotomy and 1 to 2 laminectomy was performed to improve neurologic symptoms. The posterior yellow ligament was seen partially ossified and adhered to dura without dura tear. The hematoma was found adherent to the nerve roots of the cauda equina and pia mater. After posterior segmental screw instrumentation and fusion from T10 to L3, anterior interbody fusion was performed with extrapleural approach after dissected the 11th rib, using L1 corpectomy and titanium mesh cages. There was no evidence of infection or tumor. After surgery, motor and sensory function of the lower limbs improved remarkably with solid bony union. CONCLUSION: The first case of a SSAH complicating spinal fracture in DISH is presented. The patient was successfully treated with a staged operation including posterior decompression with fusion and anterior interbody fusion. PMID- 19680096 TI - Histological changes of an injectable rhBMP-2/calcium phosphate cement in vertebroplasty of rhesus monkey. AB - STUDY DESIGN: A histologic study of recombinant human bone morphogenetic protein 2/calcium phosphate cement (rhBMP-2/CPC) using adult rhesus monkeys in vivo. OBJECTIVE: To evaluate the histologic changes of rhBMP-2/CPC in vertebroplasty and determine the feasibility of this bone substitution instead of polymethylmethacrylate (PMMA). SUMMARY OF BACKGROUND DATA: Previous studies have shown that the new rhBMP-2/nanoscale CPC has a suitable strength and injection for vertebroplasty. However, the osteoinductive properties and biodegradable characteristics are still unclear. METHODS: Percutaneous vertebroplasty (PVP) was performed in 4 adult rhesus monkeys of 2 groups. Ten vertebral bodies (VBs) from T10-L7 of each rhesus were selected, and the 20 VBs in each group were randomly divided into 3 subgroups. Subgroup A (rhBMP-2/CPC): 8 VBs, filled with rhBMP 2/CPC; Subgroup B (PMMA): 6 VBs, filled with injectable PMMA; Subgroup C (control): 6 VBs, filled with normal saline. The 2 rhesus monkeys from each of the groups were killed at 2 and 6 months after operation, respectively. Individual specimens from the 40 VBs were collected for histologic observation. RESULTS: In subgroup A, radiographic and histologic observations showed that the part of the rhBMP-2/CPC cement degraded with new bone and new vessel ingrowths, into the material, after 2 months. In addition, gaps, fibrous hyperplasia, or sclerotic callus were not found in the interface. After 6 months, the cement was nearly all replaced by mature bone tissue. In subgroup B, the inflammatory cell infiltration and fibrous membrane gapping were found after 2 months, and subsided partly at 6 months. But no new bone formation and material degradation were discovered. In subgroup C, the tunnels were filled with irregular new trabeculae after 2 months and unrecognizable from the surrounding mature bone after 6 months. CONCLUSION: It is confirmed that the rhBMP-2/CPC is an osteoinductive and biodegradable material (in animal trials). It may also be an alternative to PMMA in order to achieve biostabilization in a vertebroplasty. PMID- 19680097 TI - Biomechanical study of artificial atlanto-odontoid joint. AB - STUDY DESIGN: An in vitro biomechanical study. OBJECTIVE: To determine the initial stability and function of a new artificial joint in a cadaveric cervical spine model by comparing it with a conventional method. SUMMARY OF BACKGROUND DATA: Resection of the odontoid and anterior arch of the atlas results in atlantoaxial instability, which if left uncorrected may lead to severe neurologic complications. Currently, such atlantoaxial instability is corrected by anterior and/or posterior C1-C2 fusion. METHODS: There were 24 fresh human cadaveric cervical spines (C0-C3) randomly divided into 2 groups: group 1, resection of the odontoid with artificial atlanto-odontoid joint (AAOJ); and group 2, resection of the odontoid with Harms anterior atlantoaxial plate (Harms). For each specimen, the intact and resection of the odontoid underwent a flexibility test first, followed by the instrumented construct. Rotational angles of the C0-C3 segment were measured to study the immediate stability and function of resection of the odontoid and AAOJ, compared with the intact and resection of the odontoid and Harms. RESULTS: Compared with the intact state, resection of the odontoid and AAOJ resulted in a significant decrease in the range of motion (ROM) and neutral zone during flexion, extension, and lateral bending (P < 0.05); however, with regard to axial rotation, there was no significant difference in ROM (P > 0.05). Compared with resection of the odontoid and Harms, resection of the odontoid and AAOJ during flexion, extension, and lateral bending, there was no significant difference in ROM (P > 0.05). CONCLUSION: We have designed a new type of AAOJ for correcting atlantoaxial instability arising from C1 to C2 anterior decompression procedures. The unique aspect of this joint is that it restores, to a great extent, the C1-C2 axial rotation that is lost during current stabilization procedures. PMID- 19680098 TI - Validation of a noninvasive dynamic spinal stiffness assessment methodology in an animal model of intervertebral disc degeneration. AB - STUDY DESIGN: An experimental in vivo ovine model of intervertebral disc degeneration was used to quantify the dynamic motion response of the lumbar spine. OBJECTIVE: The purpose of this study was to: (1) compare invasively measured lumbar vertebral bone acceleration responses to noninvasive displacement responses, and (2) determine the effects of a single level degenerative intervertebral disc lesion on these responses. SUMMARY OF BACKGROUND DATA: Biomechanical techniques have been established to quantify vertebral motion responses, yet their invasiveness limits their use in a clinical setting. METHODS: Twenty-five Merino sheep were examined; 15 with surgically induced disc degeneration at L1-L2 and 10 controls. Triaxial accelerometers were rigidly fixed to the L1 and L2 spinous processes and dorsoventral (DV) mechanical excitation (20-80 N, 100 milliseconds) was applied to L3 using a spinal dynamometer. Peak force and displacement and peak-peak acceleration responses were computed for each trial and a least squares regression analysis assessed the correlation between L3 displacement and adjacent (L2) segment acceleration responses. An analysis of covariance (ANCOVA) was performed to test the homogeneity of slopes derived from the regression analysis and to assess the mean differences. RESULTS: A significant, positive, linear correlation was found between the DV displacement of L3 and the DV acceleration measured at L2 for both normal (R = 0.482, P < 0.001) and degenerated disc groups (R = 0.831, P < 0.001). The L3 DV displacement was significantly lower (ANCOVA, P < 0.001) for the degenerated group (mean: 10.39 mm) in comparison to the normal group (mean: 9.07 mm). Mean peak-peak L2-L1 DV acceleration transfer was also significantly reduced from 12.40 m/s to 5.50 m/s in the degenerated animal group (ANCOVA, P < 0.001). CONCLUSION: The findings indicate that noninvasive displacement measurements of the prone-lying animal can be used to estimate the segmental and intersegmental motions in both normal and pathologic spines. PMID- 19680099 TI - Etiology of child acute stiff neck. AB - STUDY DESIGN: The results of MRI study of 10 children with acute stiff-neck who were treated in the Department of Spine Surgery in Saint-Petersburg State Pediatric Medical Academy. OBJECTIVES: To demonstrate the MRI findings in children with acute stiff neck and suggest other explanations to this findings. SUMMARY OF BACKGROUND DATA: The etiology of child's acute stiff-neck has been debated in the published data. Most authors supported the basic role of atlantoaxial fixation in this condition. But modern investigations using MRI and CT show another explanations. METHODS: A total of 10 patients aged 5 to 14 years with typical stiff neck with acute onset were studied by MRI in first 12 hours. RESULTS: In all 10 investigations typical changes that disappeared in a few days were found. There were triangle or oblong high intensity zone near the external edges of backbone discs C2-C3 or C3-C4. The zones were always on the side where the patients felt pain. CONCLUSION: In our opinion, the main reason of the child's acute stiff-neck is a rapid or gradual strangulation of the vascularized tissue in uncovertebral zones in C2-C3, C3-C4 caused by a head movement or a neck's prolonged incurvationed position during a profound sleep. It causes a "wedge" of hydropic tissues that irritate the back longitudinal ligaments. As the result, a head has antalgic position and, in the most severe cases, is blocked. PMID- 19680100 TI - Active head and cervical range of motion: effect of age in healthy females. AB - STUDY DESIGN: A study on active cervical range of motion (RoM) in healthy females of different ages. OBJECTIVE: Perform a three-dimensional quantitative analysis of active head-cervical RoM in healthy females to assess the relationship with age. SUMMARY OF BACKGROUND DATA: Age-related variations in active cervical RoM are still partially unknown: some investigations demonstrated that age has no effect whatsoever on the primary movements, whereas other studies found an inverse proportionality between age and cervical RoM. METHODS: Three groups of females were compared: 22 aged 15 to 18 years (adolescents), 25 aged 20 to 30 years (young adults), and 16 aged 35 to 45 years (mid-aged women). Active flexion and extension, lateral bending and axial rotation were recorded via an optoelectronic system. After the mathematical exclusion of thoracic movements, cervical RoM was referred to head local reference system and calculated using the tilt/twist method. Data were compared using analysis of variance. RESULTS: Flexion and extension were larger in the adolescents (137 degrees ) than in the young adults (128 degrees ) and mid-aged women (127 degrees ). Lateral bending had similar ranges in the 3 groups: 103 degrees for adolescents, 101 degrees for young adults, and 100 degrees for mid-aged women. Axial rotation had similar ranges in the adolescents and in the mid-aged women (143 degrees ) and a slightly larger range in the young adults (151 degrees ). Primary movements were always associated with out-of-plane components. CONCLUSION: In healthy females, between 15 and 45 years old, cervical RoMs in the principal planes decrease (except for rotation), but these variations are not statistically significant (P > 0.05). PMID- 19680101 TI - 2009 updated method guidelines for systematic reviews in the Cochrane Back Review Group. AB - STUDY DESIGN: Method guidelines for systematic reviews of trials of treatments for neck and back pain. OBJECTIVE: To help review authors design, conduct and report systematic reviews of trials in this field. SUMMARY OF BACKGROUND DATA: In 1997, the Cochrane Back Review Group published Method Guidelines for Systematic Reviews, which was updated in 2003. Since then, new methodologic evidence has emerged and standards have changed. Coupled with the upcoming revisions to the software and methods required by The Cochrane Collaboration, it was clear that revisions were needed to the existing guidelines. METHODS: The Cochrane Back Review Group editorial and advisory boards met in June 2006 to review the relevant new methodologic evidence and determine how it should be incorporated. Based on the discussion, the guidelines were revised and circulated for comment. As sections of the new Cochrane Handbook for Systematic Reviews of Interventions were made available, the guidelines were checked for consistency. A working draft was made available to review authors in The Cochrane Library 2008, issue 3. RESULTS: The final recommendations are divided into 7 categories: objectives, literature search, inclusion criteria, risk of bias assessment, data extraction, data analysis, and updating your review. Each recommendation is classified into minimum criteria (mandatory) and further guidance (optional). Instead of recommending Levels of Evidence, this update adopts the GRADE approach to determine the overall quality of the evidence for important patient-centered outcomes across studies and includes a new section on updating reviews. CONCLUSION: Citations of previous versions of the method guidelines in published scientific articles (1997: 254 citations; 2003: 209 citations, searched February 10, 2009) suggest that others may find these guidelines useful to plan, conduct, or evaluate systematic reviews in the field of spinal disorders. PMID- 19680102 TI - Comparison of surgical treatment in Lenke 5C adolescent idiopathic scoliosis: anterior dual rod versus posterior pedicle fixation surgery: a comparison of two practices. AB - STUDY DESIGN: Multicenter analysis of 2 groups of patients surgically treated for Lenke 5C adolescent idiopathic scoliosis (AIS). OBJECTIVE: Compare patients with Lenke 5C scoliosis surgically treated with anterior spinal fusion with dual rod instrumentation and anterior column support with patients surgically treated with posterior release and pedicle screw instrumentation. SUMMARY OF BACKGROUND DATA: Treatment of single, structural, lumbar, and thoracolumbar curves in patients with AIS has been the subject of some debate. Advocates of the anterior approach assert that their technique spares posterior musculature and may save distal fusion levels, and that with dual rods and anterior column support the issues with nonunion and kyphosis have been obviated. Advocates of the posterior approach assert that with the change to posterior pedicle screw based instrumentation that correction and levels are equivalent, and the posterior approach avoids the issues with nonunion and kyphosis. This report directly compares the results of posterior versus anterior instrumented fusions in the operative treatment of adolescent idiopathic Lenke 5C curves. METHODS: We analyzed 62 patients with Lenke 5C based on radiographic and clinical data at 2 institutions: 31 patients treated with posterior, pedicle-screw instrumented fusions at 1 institution (group PSF); and 31 patients with anterior, dual-rod instrumented fusions at another institution (group ASF). Multiple clinical and radiographic parameters were evaluated and compared. RESULTS: The mean age, preoperative major curve magnitude, and preoperative lowest instrumented vertebral (LIV) tilt were similar in both groups (age: PSF = 15.5 years, ASF = 15.6 years; curve size: PSF = 50.3 degrees +/- 7.0 degrees , ASF = 49.0 degrees +/- 6.6 degrees ; LIV tilt: PSF = 27.5 degrees +/- 6.5 degrees , ASF = 27.8 degrees +/- 6.2 degrees ). After surgery, the major curve corrected to an average of 6.3 degrees +/- 3.2 degrees (87.6% +/- 5.8%) in the PSF group, compared with 12.1 degrees +/- 7.4 degrees (75.7% +/- 14.8%) in the ASF group (P < 0.01). At final follow-up, the major curve measured 8.0 degrees +/- 3.0 degrees (84.2% +/- 5.8% correction) in the PSF group, compared with 15.9 degrees +/- 9.0 degrees (66.6% +/- 17.9%) in the ASF group (P = 0.01). This represented a loss of correction of 1.7 degrees +/- 1.9 degrees (3.4% +/- 3.7%) in the PSF group, and 3.8 degrees +/- 4.2 degrees (9.4% +/- 10.7%) in the ASF group (P = 0.028). The LIV tilt decreased to 4.1 degrees +/- 3.4 degrees after surgery in the PSF group, and 4.5 degrees +/- 3.7 degrees in the ASF group. At final follow-up, the LIV tilt was 5.1 degrees +/- 3.5 degrees in the PSF group, and 4.5 degrees +/- 3.7 degrees in the ASF group. EBL was identical in both groups, and length of hospital stay was significantly (P < 0.01) shorter in the PSF group (4.8 vs. 6.1 days). There were no complications in either group which extended hospital stay or required an unplanned second surgery. CONCLUSION: At a minimum of 2-year follow-up, adolescents with Lenke 5C curves demonstrated statistically significantly better curve correction, less loss of correction over time, and shorter hospital stays when treated with a posterior release with pedicle screw instrumented fusion compared with an anterior instrumented fusion with dual rods for similar patient populations. PMID- 19680103 TI - Neurologic risk in growing rod spine surgery in early onset scoliosis: is neuromonitoring necessary for all cases? AB - STUDY DESIGN: Retrospective case series from a multicenter database. OBJECTIVE: To evaluate the risk of neurologic injury during growing rod surgeries and to determine whether intraoperative neuromonitoring is necessary for all growing rod procedures. SUMMARY OF BACKGROUND DATA: Although the use of growing rod constructs for early-onset spinal deformity has become a commonly accepted treatment, the incidence of neurologic events during growing rod surgeries remains unknown. METHODS: We reviewed data from a multicenter database on 782 growing rod surgeries performed in 252 patients. VEPTR devices and any constructs with rib attachments were excluded. A questionnaire was sent to all surgeons contributing cases requesting detailed information about all neurologic events associated with any growing rod surgery. RESULTS: There were 782 growing rod surgeries performed on 252 patients including 252 primary growing rod implantations, 168 implant exchanges, and 362 lengthenings. Five hundred sixty nine of 782 (73%) cases were performed with neuromonitoring. Only one clinical injury occurred in the series, resulting in an injury rate of 0.1% (1/782). This deficit occurred during an implant exchange while attempting pedicle screw placement, and resolved within 3 months. There were 2 cases with neuromonitoring changes during primary implant surgeries (0.9%, 2/231), 1 change during implant exchanges (0.9%, 1/116), and 1 neuromonitoring change during lengthenings (0.5%, 1/222). The single monitoring change that occurred during a lengthening was in a child with an intracanal tumor who also had a monitoring change during the primary surgery. There are anecdotal cases (outside this study group series) of neuromonitoring changes during simple lengthenings in children with uneventful primary implantations. CONCLUSION: Based on our study, the largest reported series of growing rod surgeries, the rate of neuromonitoring changes during primary growing rod implantation (0.9%) and exchange (0.9%) justifies the use of intraoperative neuromonitoring during these surgeries. As there were no neurologic events in 361 lengthenings in patients with no previous neurologic events, the question may be raised as to whether intraoperative neuromonitoring is necessary for simple lengthenings in these patients. However, caution should be maintained when interpreting our results as anecdotal cases of neurologic changes from simple lengthenings do exist outside of this series. PMID- 19680104 TI - Predicting outcome in acute low back pain using different models of patient profiling. AB - STUDY DESIGN: Prospective observational study of prognostic indicators, using data from a randomized, controlled trial of physiotherapy care of acute low back pain (ALBP) with follow-up at 6 weeks, 3 months, and 6 months. OBJECTIVE: To evaluate which patient profile offers the most useful guide to long-term outcome in ALBP. SUMMARY OF BACKGROUND DATA: The evidence used to inform prognostic decision-making is derived largely from studies where baseline data are used to predict future status. Clinicians often see patients on multiple occasions so may profile patients in a variety of ways. It is worth considering if better prognostic decisions can be made from alternative profiles. METHODS: Clinical, psychological, and demographic data were collected from a sample of 54 ALBP patients. Three clinical profiles were developed from information collected at baseline, information collected at 6 weeks, and the change in status between these 2 time points. A series of regression models were used to determine the independent and relative contributions of these profiles to the prediction of chronic pain and disability. RESULTS: The baseline profile predicted long-term pain only. The 6-week profile predicted both long-term pain and disability. The change profile only predicted long-term disability (P < 0.01). When predicting long-term pain, after the baseline profile had been added to the model, the 6 week profile did not add significantly when forced in at the second step (P > 0.05). A similar result was obtained when the order of entry was reversed. When predicting long-term disability, after the 6-week profile was entered at the first step, the change profile was not significant when forced in at the second step. However, when the change profile was entered at the first step and the 6 week clinical profile was forced in at the second step, a significant contribution of the 6-week profile was found. CONCLUSION: The profile derived from information collected at 6 weeks provided the best guide to long-term pain and disability. The baseline profile and change in status offered less predictive value. PMID- 19680105 TI - The Balanced Inventory for Spinal Disorders: the validity of a disease specific questionnaire for evaluation of outcomes in patients with various spinal disorders. AB - STUDY DESIGN: A prospective validation study. OBJECTIVE: To validate the Balanced Inventory for Spinal Disorders (BIS), a questionnaire concerning the extent to which pain affects perceived physical health, social life, mental health, and quality of life. The operational definitions of the items and the verbal descriptive scales were compared with corresponding items in the Short-Form 36 (SF-36), European Quality of Life Scale (EQ), and Oswestry Disability Index (ODI). SUMMARY OF BACKGROUND DATA: In validation studies, scales that intend to measure the same variable are compared. METHODS: The SF-36, EQ, ODI, and the BIS were filled in by 101 patients before surgical treatment. The comparisons were analyzed by statistical methods that take account of the nonmetric properties of ordered categorical data to obtain reliable results. The level of order consistency between BIS and comparing items, when present, was calculated. The Spearman rank-order correlation coefficient was also calculated. RESULTS: In the paired comparisons between the BIS pain scales and the other pain scales about 80% units more pairs were ordered than disordered, and the disorder was explained by the discriminating ability of the BIS back and leg pain items. The BIS and ODI items of limitation in walking were comparable, and the assessments of social limitations on the questionnaires were consistent; the disordered pairs being explained by different coverage of activities in the items. The assessments of physical and mental health on BIS were disordered, with the responses in SF-36 in favor of the BIS type of scale categories. The few items and response categories in the EQ did not discriminate the assessments. CONCLUSION: The BIS assessments can be regarded as being a valid disease-specific questionnaire that provides interpretable information regarding the impact of back end leg pain on well defined physical, social and mental aspects, and on the quality of life. PMID- 19680106 TI - The relationship between back pain and future vertebral fracture in postmenopausal women. AB - STUDY DESIGN: Cross sectional and prospective observational study in Japanese postmenopausal women. OBJECTIVE: The aim of the study was 2-fold. The first was to investigate what kind of comorbidities could be found in conjunction with back pain in Japanese postmenopausal women. The second was to investigate whether significant relationship between baseline back pain and future fracture exists or not. SUMMARY OF BACKGROUND DATA: Back pain has been reported to be associated with vertebral degeneration or vertebral fracture. However, there has been no available data that indicates the relationship between back pain and future fracture risks. METHODS: The subjects who visited their practitioner were examined for their prevalent back pain or pains in other site. Bone mineral density, body height, body weight, and serum parameter were measured at baseline, and comorbidities were investigated by interview. Fragility fractures were also assessed at baseline and then followed up with 1- to 2-year intervals. The correlation between back pain and baseline characteristics was investigated by logistic regression analysis. The hazard ratio of back pain to future vertebral fracture was estimated by multivariate Cox regression analysis. RESULTS: A total of 899 postmenopausal ambulatory women (62.5 +/- 10.3 years old) were enrolled and 81 subjects were dropped out from the study within 1 year. The remaining 818 postmenopausal women (62.1 +/- 10.3 years) were followed-up for 5.7 +/- 3.5 years. Compared to the group without pain, the group with back pain had significantly higher age, lower bone mineral densities at lumbar spine and hip, and higher number of prevalent vertebral fractures. The back pain was significantly associated with rheumatic arthritis (odds ratio [OR]: 2.01, P < 0.05), prevalent vertebral fracture (OR: 4.60, P < 0.001) and osteoporosis (OR: 2.14, P < 0.001). A total of 189 future fractures were observed, of which the most frequent was vertebral fractures (78.3%). The fact that baseline back pain was a significant risk factor for time-dependent vertebral fractures (hazard ratio: 1.62, 95% confidence interval: 1.16-2.27, P = 0.005) was demonstrated by the Cox hazards model after adjusting for traditional risk factors, such as age, bone mineral density, and prevalence of vertebral fractures. CONCLUSION: The data obtained in this study indicated that the back pain is significantly associated with osteoporosis and rheumatoid arthritis and that it can be useful predictor for future vertebral fracture risk in Japanese postmenopausal women in clinical settings. PMID- 19680107 TI - Spinal surgery in patients with end-stage renal disease undergoing hemodialysis therapy. AB - STUDY DESIGN: Case series retrospective review. OBJECTIVE: To present the surgical treatment guideline for spinal diseases with end-stage renal disease (ESRD) patients undergoing hemodialysis. SUMMARY OF BACKGROUND DATA: Treatment for spinal diseases with ESRD patients in is a special clinical challenge because of complex medical and clinical problems. METHODS: We retrospectively reviewed 12 patients who underwent spinal surgeries among patients with chronic renal failure at our hospital from May 2000 to September 2007. The medical records and radiologic findings for these patients were reviewed and concomitant medical diseases, laboratory findings, pre- and postoperative care, clinical outcomes, and complications were investigated. RESULTS: One patient died of pneumonia and sepsis 2 months after fusion surgery. Other postoperative complications included postoperative delirium in 3 patients and terminal ileitis and delayed primary spondylodiscitis in 1 patient each. There were no postoperative wound infections associated with the spinal surgery.The preoperative mean visual analogue scale score was 7.9 +/- 0.61, which improved to 2.2 +/- 1.25 at the time of final follow-up for 11 patients. Among 5 patients who underwent fusion surgery, solid bone fusion was achieved in only 3 patients and included those who underwent posterior lumbar interbody fusion with pedicle screw fixation. In 2 patients who underwent posterior lumbar interbody fusion with cage alone, solid fusion was not achieved. In 1 of 2 patients who underwent anterior cervical fusion with plating, solid fusion was achieved. The overall fusion rate was 57.1% in patients with ESRD undergoing hemodialysis. CONCLUSION: Spinal surgeries in ESRD patients undergoing hemodialysis can be performed with acceptable outcomes; however, the complication rates and mortality rates are relatively high and the fusion rate is low. To obtain a better outcome, multiple factors such as comorbid medical diseases, laboratory abnormalities, and osteoporosis should be carefully considered. PMID- 19680110 TI - The foot and ankle in sport: Modern perspectives (Part II). PMID- 19680108 TI - A simple radiological method for recognizing osteoporotic thoracic vertebral compression fractures and distinguishing them from Scheuermann disease. AB - STUDY DESIGN: Blinded. OBJECTIVE: Examination of a new radiologic interpretation technique for distinguishing osteoporotic fractures from Scheuermann disease. STUDY OF BACKGROUND DATA: Osteoporotic vertebral fractures are a major clinical problem. A variety of empirically-derived standards have been suggested for their recognition, but the accepted limits for normal variation have been somewhat arbitrary. This report presents and examines a new radiologic technique for distinguishing osteoporotic thoracic vertebral fractures from Scheuermann disease. METHODS: Vertebral body height measurements (anterior, middle, and posterior) of thoracic vertebrae T6-T10 in 44 individuals with osteoporotic fractures, 28 individuals with Scheuermann disease and a control group of 120 individuals with unaffected vertebrae, were taken from lateral chest radiographs and measured by a digital caliper. For each vertebra, 4 indexes were calculated: 3 primary derivatives; anterior height/posterior height; anterior height/mid height (A/M); mid height/posterior height (M/P); and a secondary derivate index (A/M)/(M/P). Qualitative analysis was subsequently carried out, using lateral spine radiographs of individuals with osteoporotic vertebral fractures and individuals with Scheuermann disease. Statistical analysis included analysis of variance, paired t tests and t tests. RESULTS: The second derivative (A/M)/(M/P) indexes of thoracic vertebrae T6-T10 were significantly greater in the osteoporosis group than in the control and Scheuermann groups (P < 0.05). Contrary to the control and Scheuermann groups, the M/P indexes (T6-T10) were significantly smaller than the A/M indexes in the osteoporosis group. The anterior height/posterior height indexes (T6-T10) in all 3 groups were statistically indistinguishable. Qualitative analysis showed that the intersection of an anterior-midpoint line and a posterior-midpoint line drawn on the superior vertebral body surface (T6-T10) created an "angle of depression" in the osteoporosis group whereas an "angle of elevation" was identified in the Scheuermann group. CONCLUSION: Second derivative indexes of vertebral body height parameters allow identification of vertebrae with osteoporotic compression fractures and their distinction from vertebrae with Scheuermann disease. PMID- 19680111 TI - Stress fractures of the foot and ankle. AB - Stress fractures are common athletic injuries of the foot and ankle, described in every bone except the lesser toes, and reviewed here. Early diagnosis usually allows for simpler treatment and quick recovery. Early clinical presentations can be subtle, so a high degree of suspicion and a systematic approach, coupled with an understanding of the diagnostic limitations present in early injury, is required. Such a rigorous approach ultimately pays dividends for these patients, who are usually keen to return quickly to athletic activity. "High-risk" fractures include the medial malleolus, the talus, the navicular bone, the base of the fifth metatarsal, and the hallux sesamoids. We support recommendations of early surgery in high-risk fractures. PMID- 19680112 TI - First ray disorders in athletes. AB - Athletes who participate in contact sports (American football, soccer, rugby) or who are involved in high-impact sports (dancing, running, gymnastics) are susceptible to first ray forefoot injuries. Common first ray disorders in athletes include hallux rigidus, turf toe, sand toe, sesamoid disorders, and fractures. First ray disorders in athletes frequently are treated by nonoperative methods including relative rest, ice, elevation, activity modification, shoe modification, and insoles. PMID- 19680113 TI - Disorders of the lesser toes. AB - Lesser toe disorders are an often under-appreciated source of pain and disability in athletes. Patients may have significant symptoms from corns, hammertoe and mallet toe deformities, and metatarsalgia resulting from neuromas and metatarsophalangeal joint instability. Although patients may present with vague symptoms, a careful history and physical examination will point a clinician to an accurate diagnosis. Treatment of these lesser toe disorders is straightforward and leads to predictably good results. PMID- 19680114 TI - Arthroscopy and endoscopy of the ankle and hindfoot. AB - Ankle arthroscopy provides the surgeon with a minimally invasive treatment option for a wide variety of indications such as impingement, osteochondral defects, loose bodies, ossicles, synovitis, adhesions, and instability. Posterior ankle pathology can be treated using endoscopic hindfoot portals. These posteromedial and lateral hindfoot portals provide excellent access to the posterior aspect of the ankle and subtalar joint. Also extra-articular structures in the hindfoot, for instance recurrent peroneal tendon dislocation, can be treated by creating an additional portal. The endoscopic hindfoot portals are safe and reliable, both anatomically and clinically. It compares favorably to open surgery with regard to less morbidity and a quicker recovery. PMID- 19680115 TI - Postero-medial approach procedure in the supine position for one-step anterior and posterior ankle arthroscopy. AB - Standard arthroscopy of the ankle does not allow to approach at the same time the anterior and posterior compartments. Keeping the patient supine and with the application of judicious traction, anterior ankle arthroscopy allows to examine the various intra-articular structures, but treat is limited only to anterior pathology. Also, it is not possible to explore posterior compartment and posterior hindfoot through an anterior approach. Normally, for those patients in whom both the anterior and posterior compartments were to be operated upon, surgery is stopped, and the patient has to be re-positioned. We describe a 2 postero-medial hindfoot portals procedure, which allows to reach both the posterior aspect of the ankle joint and the extra-articular compartment of the hindfoot keeping the patient supine throughout the procedure. After arthroscopy of the anterior compartment using standard anterior portals, 2 postero-medial endoscopic approaches make it possible to visualize and treat pathologies of the posterior ankle and of the hindfoot, without prolonging the operation through the need to reposition the patient in the prone position. PMID- 19680116 TI - Basic science and clinical aspects of achilles tendinopathy. AB - Achilles tendinopathy is one of the common disorders seen in athletes. It is a degenerative rather than an inflammatory, condition. There is a lifetime risk of 52% in former elite male distance runners. This condition is difficult to manage and up to 29% of Achilles tendinopathy patients may require surgery. It often results in chronic pain and discomfort along with failure to regain full function. It can be complicated by partial tears or complete rupture causing long term burden on healthcare system and making treatment quite difficult. PMID- 19680117 TI - Chronic exertional compartment syndrome of the foot. AB - Chronic exertional compartment syndrome (CECS) usually refers to myoneural ischemia from a reversible increase in tissue pressure within a myofascial compartment. CECS of the leg is well documented, as its first description by Mavor in 1956. CECS of the foot remains underdiagnosed, and has been reported in the literature only on an anecdotal basis. Wood Jones proposed that there were 4 compartments in the foot, but Manoli and Weber suggest that there are 9 separate compartments. Clinical signs and symptoms of CECS of the foot remain vague, diverse, and lack the consistency of its counterpart in the leg. The most effective treatment is a fasciotomy. We present a literature review of the condition to increase the awareness and high index of suspicion among the clinicians as the symptoms are often vague and, to consider the condition as part of the differential diagnosis. PMID- 19680118 TI - Rehabilitation and return to play after foot and ankle injuries in athletes. AB - Rehabilitation after acute ankle injury can be categorized in to early, middle, and late phases. This paper aims to cover some of the key concepts from initial management to end stage rehabilitation that could be applied to many musculoskeletal injuries around the foot and ankle. Pathology specific rehabilitation, functional performance tests, and sports specific and return to play issues are also addressed, as the aim of any rehabilitation programme is to return the athlete to the field of play as quickly as it is safe to do so. PMID- 19680119 TI - Intralymphatic immunotherapy. AB - PURPOSE OF REVIEW: IgE-mediated allergy can be treated by subcutaneous allergen specific immunotherapy (SCIT). However, the percentage of allergic patients undergoing SCIT is low, mainly due to the long duration of the therapy and the risk of severe systemic allergic reactions associated with the allergen administration. Typically, SCIT requires dozens of subcutaneous allergen injections that stretch over 3-5 years. Over the last decade, sublingual immunotherapy has been established as an alternative to SCIT, but treatment duration and dosing frequencies could not be reduced. Recently, immunotherapy by direct administration of the allergen into lymph nodes [intralymphatic immunotherapy (ILIT)] has proven a promising alternative and this method is the focus of the present review. RECENT FINDINGS: Several studies on animals and on humans have shown that direct injection into lymph nodes enhanced immune responses to protein, peptide, and naked DNA vaccines. Moreover, ILIT strongly improved allergen immunotherapy, so that the number of allergen administrations as well as the allergen dose could be reduced. As ILIT was also well tolerated, practically painless, and easy to perform, patient compliance was improved as compared with SCIT. SUMMARY: Direct ILIT into a subcutaneous lymph node markedly enhances protective immune responses, so that both the dose and the number of allergen injections can be reduced, making ILIT safer and faster than other forms of immunotherapy, and most importantly, this enhances patient convenience and compliance. PMID- 19680120 TI - Modular antigen-translocation as a novel vaccine strategy for allergen-specific immunotherapy. AB - PURPOSE OF REVIEW: The purpose of the present review is to describe recent approaches aimed at improving the treatment of allergic diseases through allergen specific immunotherapy (SIT). Special emphasis will be given to the approach based on specific targeting of the major histocompatibility complex (MHC) class II antigen-presentation pathway. RECENT FINDINGS: It is well recognized that IgE mediated allergic diseases including rhinitis, atopic eczema and allergic asthma are increasing worldwide to a pandemic dimension. The only curative treatment remains allergen-SIT, which, however, requires a long treatment time of 3-5 years with up to 80 injections to confer protection. Recent findings strongly indicate that the treatment time and the number of injections could be drastically reduced by turning immunotherapy to a true vaccination. Direct injection of allergen extracts into the inguinal lymph nodes and targeting the MHC class-II antigen presentation pathway by recombinant modular antigen-translocating vaccines have the potential to cure allergic diseases in a very short time. SUMMARY: The mechanisms of allergic inflammation can be divided into four distinct stages: T cell activation, organ-selective homing, survival/reactivation and effector functions. On the basis of this new knowledge several novel concepts aimed at treating allergic diseases have been developed. The area of allergen-SIT is experiencing exciting developments. Reciprocal regulation of effector and regulatory T cell subsets is being more and more used to develop novel strategies for immunomodulation. PMID- 19680121 TI - Total intravenous anaesthesia techniques for ambulatory surgery. AB - PURPOSE OF REVIEW: The purpose of the present review is to provide an updated discussion on the use of total intravenous anaesthesia (TIVA) for ambulatory surgery, based on results from recent studies put into the context of issues already known. RECENT FINDINGS: The current use of TIVA for ambulatory surgery seems to be abundant. It is encouraged by the simplicity of the method, increased experience and declining costs with the propofol and remifentanil combination. The TIVA methods are well tolerated and perceived to give good quality patient care; with rapid, clear-headed emergence and low incidence of postoperative nausea and vomiting. Cost-efficacy and other benefits of recovery from TIVA versus alternative techniques of anaesthesia seem to depend more on the patient and the individual perioperative setting than on the TIVA concept per se. Further development of TIVA will include the refinement of target control systems, the introduction of new drugs and adjuvants and advanced equipment for automatic drug delivery, as well as improved effect monitoring. SUMMARY: TIVA is well tolerated and simple. It is associated with less postoperative nausea and vomiting than inhalational anaesthesia and has no residual paralyses as are possible with locoregional techniques. Propofol with remifentanil seems to be the dominating TIVA technique, delivered either by conventional pumps or by target control systems. PMID- 19680122 TI - Paravertebral block: cervical, thoracic, lumbar, and sacral. AB - PURPOSE OF REVIEW: This article outlines the new developments around all four types of paravertebral block: cervical, thoracic, lumbar, and sacral. RECENT FINDINGS: It is emphasized that paravertebral blocks are all performed on the level of the roots of the nerves or plexuses, which are surrounded by dura mater extending from the dura mater surrounding the spinal cord. Because of this, they are all performed with essentially the same technique and they all have the same characteristics and problems associated with them, although they are called by many different names: 'cervical paravertebral' vs. 'posterior approach', 'psoas compartment' vs. 'lumbar plexus block' vs. 'lumbar paravertebral block', and so on. The knowledge that the roots are surrounded by dura and root level blocks are done just outside the dura should afford all paravertebral blocks the same respect as spinal epidural blocks because the potential of devastating complications such as intrathecal or intramedullary injection, for example, is shared. The advances in ultrasound and other novel approaches are described, yet the value of these has not been established. SUMMARY: We discuss commonality and differences between the four types of paravertebral blocks, and newer indications and concerns, especially pertaining to nerve microanatomical differences, are highlighted. PMID- 19680123 TI - Modification of red blood cells for laboratory quality control use. AB - PURPOSE OF REVIEW: This review describes the current state-of-the-art with respect to the modification of red blood cells for creating quality controls for use in immunohaematology. RECENT FINDINGS: The author has identified five technologies able to create modified red blood cells potentially suitable for use in quality control. Two of the technologies use enzymes, glycosidases or glycosyltransferases, to modify red blood cells and create ABO quality control cells. A third technology uses polyethylene glycol to reduce antigen expression by masking epitopes, whereas a fourth technology is speculative and involves the in-vitro generation of genetically modified erythroid cells. None of these four technologies are in routine use to make commercially available quality controls. A fifth commercially available technology creates quality controls by adding synthetic blood group A and B antigens (FSLs) to group O red blood cells, creating what are referred to as 'kodecytes'. This technology is also being used to add blood group peptides onto red cells for use in the future in a range of diagnostic applications. SUMMARY: Transducing cell-derived erythroid populations with blood group encoding or silencing vectors, and the use of FSLs to create kodecytes, are two technologies with the potential to provide quality controls for laboratory use. PMID- 19680124 TI - Trends of hematopoietic stem cell transplantation in the third millennium. AB - PURPOSE OF REVIEW: Hematopoietic stem cell transplantation (HSCT) has evolved into an accepted therapy for many congenital or acquired disorders of the hematopoietic system. It has seen major changes in indications and use of transplant techniques. HSCT is a high cost procedure and requires investments; information on trends is essential for patient counselling and healthcare planning. RECENT FINDINGS: HSCT rates have increased worldwide. Increase is constant and predictable; the reasons therefore are manifold. Introduction of reduced intensity conditioning regimens has opened access to patients at older age and with comorbidities. Higher numbers of unrelated volunteer donors and cord blood products give access to HSCT for patients without family donors. For some well defined indications, HSCT has become the most cost efficient therapy in countries with limited resources. Use of HSCT is under discussion for nonhematopoietic indications, as is the use of nonhematopoietic stem cells for organ repair. SUMMARY: HSCT is likely to continue to increase for currently established indications. Indications and technologies will vary between countries with limited or nonrestricted resources. The most cost effective approach might not be the same everywhere. Novel indications will emerge but time will be needed to confirm their benefit. Close observation of global trends will become an essential tool for healthcare agencies in order to provide the necessary infrastructure in time. PMID- 19680125 TI - Hereditary predominantly motor neuropathies. AB - PURPOSE OF REVIEW: We review recent advances in Charcot-Marie-Tooth disease (CMT), the most frequent inherited neuromuscular disorder. RECENT FINDINGS: During the last year further progresses have occurred in this field and concerned identification of novel mutations in recently identified genes, allowing better definition of associated phenotypes; increased knowledge on pathophysiologic mechanisms of the different CMT types, with the contribution of cellular and animal model studies; studies on the natural history of CMT and attempts at developing appropriate outcome measures to assess disease course and intervention efficacy; trials with ascorbic acid in CMT type 1A; and studies on new possible therapeutic strategies. SUMMARY: Such advances have implications on clinical management of CMT and are modifying the clinical approach to CMT, by improving diagnostic tools, allowing better definition of prognosis, and increasing the hope for future effective treatments. Research on CMT is important as is shedding light on important pathways that regulates the normal function of axonal transport, vesicular trafficking, and also revealing new aspects of intracellular organelles' function and interactions. PMID- 19680126 TI - Inflammatory myopathies: disease mechanisms. AB - PURPOSE OF REVIEW: Recent developments pertaining to disease mechanisms in the inflammatory myopathies are discussed, emphasizing those areas that are of particular interest to me. RECENT FINDINGS: The identification and further characterization of the type 1 interferon pathway in dermatomyositis is leading down a path of genomic medicine. Myonuclear structural abnormalities and the presence of nucleic acid-binding proteins, including the TAR DNA binding protein TDP-43, in sporadic inclusion body myositis (sIBM) sarcoplasm are important recent observations. This is an area likely to provide deep understanding of the mechanism of myofiber injury in sIBM. Proteomic characterization of proteins in sIBM muscle, muscle functioning as a lymphoid tissue, and the nature of belief systems, particularly one pertaining to beta-amyloid and sIBM, are other areas of interest. SUMMARY: Clarification of disease mechanisms is providing a basis for rational drug development for some patients with myositis. PMID- 19680127 TI - Agents and mechanisms of toxic myopathy. AB - PURPOSE OF REVIEW: Mechanistic-based research has made possible a more pathophysiologic approach to certain drug-induced muscle disorders, especially those caused by the lipid-lowering statin family of drugs, but also myopathies caused by antimicrotubule drugs, mitochondrial toxins, foods, and purported nutriceutical remedies. This is a critical review of those syndromes that are most well founded on evidence of challenge/de-challenge/re-challenge, case controls, or experimental controls. RECENT FINDINGS: Statins are well tolerated drugs with very high safety windows in skeletal muscle, and third-generation statins now under development offer the hope of even less risk of toxic myopathy. Toxicity is dose-related and time-related, and is due to intramyofiber cascades downstream from 3-hydroxy-3-methylglutaryl-coenzyme A (HMG Co-A) reductase inhibition. A robust pathophysiologic animal model shows that statins decrease strength and increase cytosolic Ca2+ by increasing both mitochondrial Ca2+ permeability and Ca2+ release from sarcoplasmic reticulum. As a result, the earliest pathologic change in statin myotoxicity is compatible with simple necrosis and intracellular membrane accumulation. Genome-wide searching has yielded a single nucleotide polymorphism in the SLCO1B1 gene for the organic anion-transporting polypeptide that regulates statin uptake. Drug-drug interactions dominate recent reports of all toxic myopathies. The peculiar mitochondrial pathology of zidovudine-induced mitochondrial DNA depletion, cytochrome oxidase depletion, and mitochondrial proliferation has been confirmed in a rigorous animal model. Finally, recent interest has been piqued by putative lipid-lowering neutraceuticals like red yeast rice (Monascus purpureus) and edible mushrooms that can clearly cause toxic myopathy. SUMMARY: A principled approach to the diagnosis of toxic myopathies, based on the consideration of currently known pathophysiologic mechanisms, biopsy pathology, the characteristic clearance properties of creatine kinase, the time course of muscle fiber regeneration, drug challenge/de-challenge/re-challenge, and differential diagnoses, rather than on mere temporal association, will reduce the healthcare costs of common diagnostic error. PMID- 19680128 TI - Reconstructive urology and geriatrics: issues in older adults. AB - PURPOSE OF REVIEW: The elderly population is continuing to expand at record pace in the vast majority of countries worldwide. Many urologic conditions that necessitate reconstructive surgery occur in geriatric patients. To date, there has been a paucity of research on the effects of aging with regard to reconstructive procedures in elderly patients. This review examines factors that influence the feasibility and outcomes of reconstructive urologic surgery in older adults. RECENT FINDINGS: Age alone has generally not been identified as a significant predictive factor for outcomes in geriatric patients undergoing surgery. Comorbid diseases associated with reduction of physiologic reserve capacity and impairments in the level of independence for activities of daily living appear to have more predictive value. Recent research has focused on defining frailty as a condition that frequently occurs in older adults and may influence clinical outcomes. A variety of cellular and tissue changes associated with aging have also been studied. Inflammatory mediators may play an important role in this process. SUMMARY: Many urologic conditions that require reconstructive surgery occur in elderly patients. Careful planning may help to improve outcomes. However, there appear to be inherent changes associated with the physiology of normal aging that can significantly influence this process. PMID- 19680129 TI - Ureteric reconstruction and replacement. AB - PURPOSE OF REVIEW: To review the recent advances on ureteric reconstruction and replacement, in particular, ileal ureteric replacement and laparoscopic and robotic-assisted ureteral reconstruction. RECENT FINDINGS: Recently, the ureteric replacement with bowel has been carefully assessed by several authors, and the results are quite impressive. Also, very recent studies on laparoscopic and robotic-assisted ureteral repair have been published. Outcomes appear very promising, allowing for a faster recovery and shorter hospital stay for the patient. SUMMARY: Today, we can conclude that the field of ureteric reconstruction and replacement is still evolving. Old techniques are supported by an increasing degree of evidence, and new, more minimally invasive surgical strategies emerge. Clearly, there are some disadvantages as well as difficulties to overcome with the new techniques; however, recent studies appear to present promising results. PMID- 19680130 TI - Computer simulator among experts involved in screening colonoscopy. AB - BACKGROUND: In the era of screening colonoscopy, assessment of operator competence is warranted. AIM: To evaluate feasibility of a computer simulator (CS) use for assessment of competence in colonoscopy by investigating performance of expert endoscopists at CS. SUBJECTS: Twenty expert endoscopists involved in screening colonoscopy. METHODS: Experts returned a questionnaire regarding personal practice (duration of activity, number of colonoscopies in the last year and assistance by a nurse) and performances (percentage of caecal intubation and polyp detection rate). One easy and one difficult colonoscopy were proposed at CS in randomized order. RESULTS: Participation rate was 75%. Caecal intubation rate in clinical practice was more than 90% for all experts. At CS, time to caecal intubation and number of attempts for ileal intubation were significantly lower during easy versus difficult colonoscopy (P<0.01 for both items); interestingly, percentage of mucosa explored was higher (P<0.05) during the difficult simulation. Withdrawal time >/=6 min was achieved by 40 and 33% of experts during the easy and difficult simulation, respectively. Independent of simulation difficulty, time with loop was lower (P<0.05) for experts using hands-free insertion (n = 8) compared with those using nurse assistance in their clinical practice (n = 7). No correlation was found between scores at CS and performance in clinical practice. CONCLUSION: Scores at CS are sensitive to the rate of technical difficulty and nurse assistance during daily practice. Withdrawal time is often shorter than required for high accuracy in polyp detection. CS could be a well-accepted tool for assessment of competence. PMID- 19680132 TI - Transcatheter closure of coronary artery fistula using the new Amplatzer vascular plug and a telescoping catheter technique. AB - Coronary artery fistulas (CAFs) are rare congenital or acquired anomalies characterized by an abnormal communication between the coronary arteries and the cardiac chambers or great vessels. Most patients are asymptomatic during childhood but symptoms and complications have been reported with advancing age. Until recently, surgery was the routine mode of treatment for CAFs but, today transcatheter closure is recommended using a variety of devices, such as occlusion coils, vascular plugs, umbrella devices and covered stents. The case described here is of a 47-year-old woman with a large bilateral CAF draining into the pulmonary artery, successfully treated by implantation of two Amplatzer vascular plugs using a telescoping catheter technique. PMID- 19680133 TI - Implantable cardioverter-defibrillator lead infection detected by 99mTc-sulesomab single-photon emission computed tomography/computed tomography 'fusion' imaging. AB - We describe a case of implantable cardioverter-defibrillator (ICD) lead infection that was diagnosed by means of sulesomab imaging whereas ultrasound examinations were inconclusive. Nuclear techniques may be helpful in such, not infrequent, settings. PMID- 19680131 TI - The acute effects of HIV protease inhibitors on insulin suppression of glucose production in healthy HIV-negative men. AB - BACKGROUND: The effects of different HIV protease inhibitors (PIs) on peripheral insulin resistance have been described, but less is known about their effects on insulin suppression of endogenous glucose production (EGP). METHODS: We tested the acute effects of 3 PIs, indinavir, ritonavir, and amprenavir, on EGP quantified by stable isotope techniques during the hyperinsulinemic, euglycemic clamp in 3 similar placebo-controlled protocols. RESULTS: EGP was higher with indinavir in the hyperinsulinemic state than with placebo (4.1 +/- 1.3 vs. 2.2 +/ 0.8 microg x kg(-1) x min(-1), P = 0.04). A trend toward higher EGP was seen with ritonavir (3.6 +/- 0.3 vs. 3.0 +/- 0.5 microg x kg(-1) x min(-1), P = 0.08). There was no evidence that amprenavir blunted insulin suppression of EGP compared with placebo (2.9 +/- 0.04 vs. 3.2 +/- 0.7 microg x kg(-1) x min(-1), P = 0.63). CONCLUSIONS: Some PIs can acutely blunt the ability of insulin to suppress EGP, but, as with insulin resistance, the effects of PIs on EGP are drug-specific, not class-specific. PMID- 19680134 TI - Interview versus questionnaire symptom reporting in people with the postconcussion syndrome. AB - OBJECTIVE: To compare spontaneous, interview-based, postconcussion symptom reporting to endorsement of symptoms on a standardized questionnaire. PARTICIPANTS: Sixty-one patients referred to a concussion clinic following mild traumatic brain injury. PROCEDURE: Patients recalled their current symptoms and problems via open-ended interview and then completed a structured postconcussion checklist. MAIN OUTCOME MEASURES: Open-ended interview and the British Columbia Postconcussion Symptom Inventory (BC-PSI). RESULTS: On average, patients endorsed 3.3 symptoms (SD = 1.9) during open-ended interview and 9.1 symptoms (SD = 3.2) on the BC-PSI (P < .001). Approximately 44% endorsed 4 or more symptoms during interview compared with 92% on the BC-PSI. The percentage of patients endorsing items on the BC-PSI compared with interview was significantly greater on all 13 items. It was common for patients to endorse symptoms as moderate-severe on the BC-PSI, despite not spontaneously reporting those symptoms during the interview. CONCLUSIONS: Clinicians need to be cautious when interpreting questionnaires and be aware of the possibility of nonspecific symptom endorsement, symptom overendorsement, symptom expectations influencing symptom endorsement, and the nocebo effect. PMID- 19680135 TI - Stimulation of AT2 receptor exerts beneficial effects in stroke-prone rats: focus on renal damage. AB - BACKGROUND AND AIM: Angiotensin II acts through two major receptors: AT1-R and AT2-R. It is known that the stimulation of AT1-R mediates vasoconstriction, cell proliferation and fibrosis, aldosterone release and inflammatory response but, although the stimulation of AT2-R is thought to promote vasodilation and anti inflammatory effects, its real in-vivo functions are still unclear. The aim of this study was to investigate the effects of specific and selective AT2-R stimulation on the pathological events occurring in spontaneously hypertensive stroke-prone rats (SHRSPs). METHODS AND RESULTS: SHRSPs who were fed a high-salt diet underwent long-term treatment with vehicle or compound 21 (C21), a nonpeptide selective AT2-R agonist, at doses of 0.75, 5 and 10 mg/kg per day. The vehicle-treated rats developed brain abnormalities detectable by magnetic resonance imaging after 42.5 +/- 7.5 days, and died 43 +/- 9.5 days after the start of the dietary treatment. The highest C21 dose delayed the occurrence of brain damage (P < 0.001 vs. vehicle-treated SHRSPs) and prolonged survival (P < 0.001) without affecting blood pressure. These beneficial effects of C21 were abolished by the administration of PD123319, an AT2-R antagonist. C21 treatment preserved renal structure by preventing inflammatory cell infiltration, collagen accumulation, and the neo-expression of vimentin; it also prevented the increased plasma renin activity and accumulation of urinary acute-phase proteins observed in the vehicle-treated rats. CONCLUSION: Specific and selective AT2-R stimulation has beneficial effects on the pathological events occurring in SHRSPs. These data indicate a new avenue for the pharmacological treatment of diseases in which modulation of the renin-angiotensin system is required. PMID- 19680137 TI - Adenine/cytosine(1166) polymorphism of the angiotensin II type 1 receptor gene and the antihypertensive response to angiotensin-converting enzyme inhibitors. AB - BACKGROUND: Most of the known actions of angiotensin II are mediated by the angiotensin II type 1 receptor (AGT1R). The adenine/cytosine(1166) (A/C(1166)) polymorphism of the AGT1R gene has been shown to be associated with hypertension and hypertension-related diseases. Thus, it may have the potential to predict the blood pressure response of patients with hypertension to angiotensin-converting enzyme inhibitors (ACEIs). OBJECTIVE: To investigate the association between the A/C(1166) polymorphism and the blood pressure response to ACEIs in a hypertensive cohort. DESIGN AND METHODS: After a 2-week, single-blind, placebo run-in period, ACEIs were administered for 6 weeks to 509 hypertensive patients. The polymorphism was determined by PCR followed by restriction enzyme digestion. RESULTS: The AA genotype, AC genotype, and CC genotype were present in 464 (91.2%), 44 (8.6%), and 1 (0.2%) of patients, respectively. As the frequency of the C allele was quite low (0.05), the genotypes were classified according to the presence or absence of the C allele. After 6 weeks of treatment, the systolic blood pressure reductions in patients with the AA genotype and AC + CC genotype were 14.3 +/- 12.6 and 14.1 +/- 12.2 mmHg, respectively (P = 0.908). The diastolic blood pressure reductions in patients with the AA genotype and AC + CC genotype were 8.6 +/- 7.1 and 8.8 +/- 6.9 mmHg, respectively (P = 0.816). There were no differences in the changes in systolic or diastolic blood pressure after treatment among the two genotype groups. CONCLUSION: The AGT1R A/C(1166) polymorphism does not predict the response to antihypertensive treatment with ACEIs in Chinese hypertensive patients. PMID- 19680136 TI - Retinal vessel diameters and risk of hypertension: the Multiethnic Study of Atherosclerosis. AB - OBJECTIVE: To describe the prospective relationship of retinal vessel diameters with risk of hypertension in a multiethnic population-based cohort. METHODS: The Multi-Ethnic Study of Atherosclerosis is a population-based study of subclinical cardiovascular disease among white, African-American, Hispanic, and Chinese American adults aged 45-84 years. Retinal vessel diameters were measured using a standardized imaging software at the second examination (considered baseline in this analysis) and summarized as the central retinal artery/vein equivalent. Presence of retinopathy and retinal focal arteriolar narrowing and arteriovenous nicking was assessed by trained graders. Incidence of hypertension was defined among participants at risk as systolic blood pressure at least 140 mmHg, diastolic blood pressure at least 90 mmHg, or use of an antihypertensive medication. RESULTS: Of the initial 6237 participants at baseline, 2583 were at risk of hypertension. After 3.2 +/- 0.5 years of follow-up, 448 (17.3%) participants developed hypertension. After adjusting for age, sex, race/ethnicity, the average of mean arterial blood pressure in the first and second examination, and other vascular risk factors, persons with narrower retinal arteriolar diameter and wider venular diameter at baseline were more likely to develop hypertension [odds ratio per SD decrease in central retinal artery equivalent 1.20, 95% confidence intervals 1.02, 1.42; and odds ratio per SD increase in central retinal vein equivalent 1.18, 95% confidence interval 1.02, 1.37]. Persons with focal arteriolar narrowing were also more likely to develop hypertension (odds ratio 1.80, 95% confidence interval 1.09, 2.97). CONCLUSION: Findings from this multiethnic population confirm that narrower retinal arteriolar diameter and wider venular diameter are associated with the development of hypertension independent of traditional risk factors. PMID- 19680138 TI - Cardiovascular and renal toxicity during angiogenesis inhibition: clinical and mechanistic aspects. AB - Inhibition of angiogenesis with humanized monoclonal antibodies to vascular endothelial growth factor (VEGF) or with tyrosine kinase inhibitors targeting VEGF receptors has become an established treatment for various tumor types. Contrary to expectations, angiogenesis inhibition by blocking VEGF-mediated signaling is associated with serious side effects including hypertension and renal and cardiac toxicity in a substantial proportion of patients. Fortunately, most of these side effects as discussed in this paper seem to be manageable, but likely become more problematic when survival increases. Although several hypotheses regarding the etiology of angiogenesis inhibition-related cardiovascular and renal side effects have been postulated, many of the underlying pathophysiological mechanisms remain to be elucidated. This may lead to the development of more specific angiogenesis inhibitors, better management of their side effects and may potentially provide new insights into the pathogenesis of cardiovascular disease in general. PMID- 19680140 TI - Selective deposition of 4-repeat tau in cerebral infarcts. AB - The tau deposits found in neurodegenerative diseases are classified based on their isoforms, that is, 3-repeat (3R) tau and 4-repeat (4R) tau. These isoforms are distinguishable using the antibodies RD3 and RD4, respectively, and Gallyas (Gal) and Campbell-Switzer (CS) silver staining methods, respectively. Tau is also deposited in cerebral infarcts. To characterize the tau profile in these lesions, 21 brains from autopsied patients with cerebral infarcts were analyzed using immunohistochemistry with RD3, RD4, and the anti-paired helical filament antibody AT8 and with Gal and CS staining; all of these techniques identify Alzheimer disease-type neurofibrillary tangles. Fluorescence labeling followed by silver staining in mirror-section pairs was also used to compare the staining patterns. Neurons in and around ischemic foci exhibited the 4R-tau epitope until 34 days postinfarction; argyrophilia with Gal staining persisted longer. The 4R tau/Gal-positive neurons were negative for 3R-tau and AT8 epitopes and lacked fibrillary structures and argyrophilia by CS staining; they are, therefore, distinct from neurons with neurofibrillary tangles. Positivity for 4R tau/Gal and negativity for 3R tau/CS were also seen in astrocytes and microglia around infarcts. Although this staining profile is characteristic of degenerative processes with 4R-tau deposition, lack of AT8 immunoreactivity and of fibrillary structures in neurons, astrocytes, and microglia indicates that selective 4R-tau deposition represents a stage without tau phosphorylation or fibril formation in cerebral infarcts. PMID- 19680141 TI - Meningeal inflammation is not associated with cortical demyelination in chronic multiple sclerosis. AB - Cortical demyelination can be extensive in chronic multiple sclerosis (MS) patients. Cortical lesions are not associated with lymphocyte infiltration, blood brain barrier disruption, or complement deposition; therefore, their pathogenesis is unclear. We analyzed the extent and cellular composition of leptomeningeal inflammatory infiltrates and their relationship with cortical demyelinated lesions in brain autopsy samples from 28 chronic MS patients; samples from 6 nonneurological disease control patients were also studied. Immunohistochemistry was used to detect meningeal T cells, B cells, macrophages, mature and immature dendritic cells, T-helper cells, (activated) cytotoxic T cells, and plasma cells. Quantitative analysis revealed significant meningeal inflammation in chronic MS patients; T cells were the predominant inflammatory cells. Morphometric analysis was performed on coronal hemisphere sections of the MS cases to assess subpial demyelination; no correlation between the extent of subpial demyelination and extent of meningeal inflammation was identified. Moreover, no differences were observed in the degree or cellular composition of meningeal infiltrates in areas directly adjacent to subpial lesions compared with areas adjacent to normal appearing gray matter in the MS cases. In addition, no follicle-like structures were found in the MS samples. Our data suggest that the occurrence of cortical lesions is not related to the presence of meningeal inflammation in a large number of chronic MS patients. PMID- 19680139 TI - Demyelinated axons and motor function are protected by genetic deletion of perforin in a mouse model of multiple sclerosis. AB - Axon injury is a major determinant of the loss of neurological function in patients with multiple sclerosis. It is unclear, however, whether damage to axons is an obligatory consequence of demyelination or whether it is an independent process that occurs in the permissive environment of demyelinated lesions. Previous investigations into the role of CD8 T cells and perforin in the Theiler murine encephalomyelitis virus model of multiple sclerosis have used mouse strains resistant to Theiler murine encephalomyelitis virus infection. To test the role of CD8 T cells in axon injury, we established a perforin-deficient mouse model on the H-2 major histocompatibility complex background thereby removing confounding factors related to viral biology in this Theiler murine encephalomyelitis virus-susceptible strain. This permitted direct comparison of clinical and pathological parameters between perforin-competent and perforin deficient mice. The extent of demyelination was indistinguishable between perforin-competent and perforin-deficient H-2 mice, but chronically infected perforin-deficient mice exhibited preservation of motor function and spinal axons despite the presence of spinal cord demyelination. Thus, demyelination is necessary but insufficient for axon injury in this model; the absence of perforin protects axons without impacting demyelination. These results suggest that perforin is a key mediator of axon injury and lend additional support to the hypothesis that CD8 T cells are primarily responsible for axon damage in multiple sclerosis. PMID- 19680142 TI - Disruption of the neurogenic niche in the subventricular zone of postnatal hydrocephalic hyh mice. AB - Neural stem cells persist after embryonic development in the subventricular zone (SVZ) niche and produce new neural cells during postnatal life; ependymal cells are a key component associated with this neurogenic niche. In the animal model of human hydrocephalus, the hyh mouse, the ependyma of the lateral ventricles is progressively lost during late embryonic and early postnatal life and disappears from most of the ventricular surface throughout its life span. To determine the potential consequences of this loss on the SVZ, we characterized the abnormalities in this neurogenic niche in hyh mice. There was overall disorganization and a marked reduction of proliferative cells in the SVZ of both newborn and adult hyh hydrocephalic mice in vivo; neuroblasts were displaced to the ventricular surface, and their migration through the rostral migratory stream was reduced. The numbers of resident neural progenitor cells in hyh mice were also markedly reduced, but they were capable of proliferating, forming neurospheres, and differentiating into neurons and glia in vitro in a manner indistinguishable from that of wild-type progenitor cells. These findings suggest that the reduction of proliferative activity observed in vivo is not caused by a cell autonomous defect of SVZ progenitors but is a consequence of a reduced number of these cells. Furthermore, the overall tissue disorganization of the SVZ and displacement of neuroblasts imply alterations in the neurogenic niche of postnatal hyh mice. PMID- 19680143 TI - Abnormal localization of leucine-rich repeat kinase 2 to the endosomal-lysosomal compartment in lewy body disease. AB - Missense mutations in the leucine-rich repeat kinase 2 (LRRK2) gene are the most common causes of both familial and sporadic forms of Parkinson disease and are also associated with diverse pathological alterations. The mechanisms whereby LRRK2 mutations cause these pathological phenotypes are unknown. We used immunohistochemistry with 3 distinct anti-LRRK2 antibodies to characterize the expression of LRRK2 in the brains of 21 subjects with various neurodegenerative disorders and 7 controls. The immunoreactivity of LRRK2 was localized in a subset of brainstem-type Lewy bodies (LBs) but not in cortical-type LBs, tau-positive inclusions, or TAR-DNA-binding protein-43-positive inclusions. The immunoreactivity of LRRK2 frequently appeared as enlarged granules or vacuoles within neurons of affected brain regions, including the substantia nigra, amygdala, and entorhinal cortex in patients with Parkinson disease or dementia with LBs. The volumes of LRRK2-positive granular structures in neurons of the entorhinal cortex were significantly increased in dementia with LBs brains compared with age-matched control brains (p < 0.05). Double immunolabeling demonstrated that these LRRK2-positive granular structures frequently colocalized with the late-endosomal marker Rab7B and occasionally with the lysosomal marker, the lysosomal-associated membrane protein 2. These results suggest that LRRK2 normally localizes to the endosomal-lysosomal compartment within morphologically altered neurons in neurodegenerative diseases, particularly in the brains of patients with LB diseases. PMID- 19680145 TI - Acute and chronic 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine administrations elicit similar microglial activation in the substantia nigra of monkeys. AB - Increasing evidence suggests a pivotal role for neuroinflammation in the pathogenesis of Parkinson disease, but whether activated microglia participate in disease progression remains unclear. To clarify this issue, we determined the numbers of activated microglial cells in the substantia nigra pars compacta and ventral tegmental area of monkeys subacutely and chronically exposed to 1-methyl 4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Monkeys in the subacute MPTP treatment group were killed 1 week after the last MPTP injection; chronically treated monkeys were killed either 6 or 35 months after the last MPTP injection. Subacute MPTP administration induced loss of dopaminergic neurons in the substantia nigra pars compacta and ventral tegmental area and microglial activation in the same areas. Chronic MPTP treatment resulted in greater dopaminergic neuron depletion in both treatment groups. Both groups of chronic MPTP-treated monkeys showed increased numbers of activated microglial cells in the substantia nigra pars compacta that were similar to those of the subacute MPTP treatment group. These results indicate that microglial activation seems to be induced mainly by the toxic effects of MPTP and that it does not further progress once the toxin administration has been terminated. This suggests that the progressive degeneration of nigral cells in Parkinson disease may not necessarily be associated with progressively increased microglial activation. PMID- 19680146 TI - Preferential expression of truncated isoforms of FOXP1 in primary central nervous system lymphoma. AB - Forkhead box P1 (FOXP1) protein is a transcription factor involved in cell signaling and regulation of gene expression. The overexpression of FOXP1 in a subgroup of systemic diffuse large B-cell lymphomas has been associated with an exceptionally poor clinical outcome. Data on FOXP1 expression in primary central nervous system lymphomas (PCNSL), that is, diffuse large B-cell lymphomas confined to the central nervous system, are not yet available. We analyzed 43 PCNSL from immunocompetent patients. Immunohistochemistry showed expression of FOXP1 protein in 21 (88%) of 24 cases. All 19 PCNSL analyzed by quantitative gene expression analysis showed overexpression of truncated FOXP1 Isoforms 3 and 9 and downregulation of normal-size FOXP1 compared with nonmalignant germinal center B cells, the normal counterpart of PCNSL tumor cells. Thus, truncated FOXP1 isoforms are preferentially overexpressed in PCNSL as they are in diffuse large B cell lymphomas. Although the mechanisms are presently unclear, this overexpression may contribute to a poor prognosis in PCNSL. PMID- 19680144 TI - Peripheral neuropathy in rats exposed to dichloroacetate. AB - The use of dichloroacetate (DCA) for treating patients with mitochondrial diseases is limited by the induction of peripheral neuropathy. The mechanisms of DCA-induced neuropathy are not known. Oral DCA treatment (50-500 mg/kg per day for up to 16 weeks) induced tactile allodynia in both juvenile and adult rats; concurrent thermal hypoalgesia developed at higher doses. Both juvenile and adult rats treated with DCA developed nerve conduction slowing that was more pronounced in adult rats. No overt axonal or glial cell abnormalities were identified in peripheral nerves or spinal cord of any DCA-treated rat, but morphometric analysis identified a reduction of mean axonal caliber of peripheral nerve myelinated fibers. Dichloroacetate treatment also caused accumulation of oxidative stress markers in the nerves. These data indicate that behavioral, functional, and structural indices of peripheral neuropathy may be induced in both juvenile and adult rats treated with DCA at doses similar to those in clinical use. Dichloroacetate-induced peripheral neuropathy primarily afflicts axons and involves both metabolic and structural disorders. The DCA-treated rat may provide insight into the pathogenesis of this peripheral neuropathy and facilitate development of adjuvant therapeutics to prevent this disorder that currently restricts the clinical use of DCA. PMID- 19680147 TI - c-Jun N-terminal kinase pathway activation in human and experimental cerebral contusion. AB - The c-Jun N-terminal kinase (JNK) pathway is involved in cell stress and apoptosis. We tested the hypothesis that this pathway plays a role in traumatic brain injury (TBI) by assessing JNK activation in human brain tissues and in brains of mice subjected to controlled cortical impact brain injury. We also assessed the effects of specific inhibition of the JNK pathway by the cell permeable JNK inhibitor peptide, D-JNKI1, on neurobehavioral function and posttraumatic cell loss in mice. The inhibitor was administered intraperitoneally 10 minutes after injury. The JNK pathway showed robust activation both in human contusion specimens and in injured cortex and hippocampi of TBI-injured mice, 1, 4, and 48 hours after injury. D-JNKI1 treatment significantly improved motor performance at 48 hours and 7 days after injury and reduced the contusion volume compared with saline treatment; the numbers of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling-positive cells were significantly decreased in the hippocampi of injured mice 48 hours after treatment. Thus, because the JNK pathway is activated after human and experimental TBI and the inhibitor peptide D-JNKI1 affords significant neuroprotection and amelioration of neurobehavioral deficits after experimental TBI, therapeutic targeting of the JNK activation pathway may hold promise for future clinical applications. PMID- 19680149 TI - Evidence-based practice improvement: merging 2 paradigms. AB - This article presents a new model, Evidence-Based Practice Improvement, for improving patient care. The model merges 2 extant paradigms currently used for quality improvement initiatives-evidence-based practice and practice or performance improvement. The literature expounds on the virtues of each of these approaches, yet no authors have moved beyond parallel play between them. The merged model, Evidence-Based Practice Improvement, may provide a more effective and practical approach to reach our quality goals. PMID- 19680150 TI - Crohn disease and mycobacterial infection in children: an intriguing relationship. AB - The relation between Crohn disease (CD) and mycobacterial infection is complex and intriguing. The 2 conditions share common immunopathogenic features, and the hypothesis that Mycobacterium avium subspecies paratuberculosis may cause CD is still under debate. Moreover, differential diagnosis between CD and intestinal tuberculosis (TB) may be challenging, especially in the growing proportion of children immigrating from countries with a high prevalence of TB. Thus, in Western countries, paediatricians who are involved in the diagnosis and management of children with CD are expected to face intestinal TB more frequently than in the past and should always consider the differential diagnosis between these 2 conditions. In addition, the use of biological agents in the treatment of CD, which may reactivate latent TB, requires the development of targeted diagnostic algorithms. Children with CD who are candidates for treatment with antitumour necrosis factor-alpha agents should be screened for latent or active TB and closely followed over time, also considering possible failure of antitubercular prophylaxis and the frequent uncharacteristic presentation of TB in children receiving antitumour necrosis factor-alpha agents. In the present review, the most recent literature findings on these topics are reported, focusing particularly on the paediatric age group. PMID- 19680148 TI - Mitochondrial morphogenesis, distribution, and Parkinson disease: insights from PINK1. AB - The etiology of Parkinson disease (PD) has been assumed to be a complex combination of environmental factors, intrinsic cellular metabolic properties, and susceptible genetic alleles. The primary obstacles to the development of a neuroprotective therapy in PD include uncertainties with regard to the precise cause(s) of neuronal dysfunction and what to target. The discoveries of Mendelian genes associated with inherited forms of PD in the last 10 years have revolutionized the understanding of the cellular pathways leading to neuronal dysfunction. Common themes of the pathogenesis of PD are beginning to emerge with mitochondrial dysfunction at the center stage. In this review, we summarize our knowledge of the pathogenesis of PD, revisit some aspects of mitochondrial biology, and discuss the insights from the study of Pink1, a familial PD associated gene. We propose that mitochondrial morphogenesis and distribution might be a novel and potential common paradigm for PD and other neurodegenerative disease research and that modulation of such mitochondrial processes may prove to be a valuable therapeutic avenue for PD. PMID- 19680151 TI - Endoscopic retrograde cholangiopancreatography in patients with neonatal cholestasis: an additional diagnostic tool for selected indications. PMID- 19680152 TI - Selective use of endoscopic retrograde cholangiopancreatography in the diagnosis of biliary atresia in infants younger than 100 days. AB - OBJECTIVES: We investigated the role and safety of endoscopic retrograde cholangiopancreatography (ERCP) in diagnosing biliary atresia (BA) in prolonged neonatal cholestasis, when standard workup was inconclusive. PATIENTS AND METHODS: We reviewed notes of 48 cholestatic infants younger than 100 days undergoing ERCP from 1997 to 2007. RESULTS: Amongst approximately 3300 infants evaluated for liver disease during the study, 224 (6.8%) were diagnosed with BA. Forty-eight children underwent ERCP. Findings at liver biopsy (n=47) included nonspecific cholestasis (n=19, 40%), giant-cell hepatitis (n=12, 26%), "large bile duct obstruction" (n=9, 19%) in the presence of pigmented stools, and mixed cholestatic/hepatitic features (n=7, 15%). ERCP demonstrated a patent biliary tree in 20 infants (42%). BA was confirmed at exploratory laparotomy in all 3 infants (6%) in whom cannulation failed. The remaining 25 infants (52%) also proceeded to exploratory laparotomy, in which BA was confirmed in 22 (46%). Amongst the 20 children in whom ERCP ruled out BA, 8 (17%) had normal biliary anatomy, whilst 12 (25%) had an abnormal biliary tree, including 6 (12.5%) with neonatal sclerosing cholangitis. After ERCP none developed clinical pancreatitis or peritonitis. CONCLUSIONS: ERCP is a safe procedure for diagnosing BA even in the smallest infants with high positive and negative predictive values. PMID- 19680153 TI - Is familial Mediterranean fever a possible cofactor for Budd-Chiari syndrome? PMID- 19680154 TI - Comparison of esophageal pH and multichannel intraluminal impedance testing in pediatric patients with suspected gastroesophageal reflux. AB - BACKGROUND: Multichannel intraluminal impedance (MII) is a pH-independent method of assessing gastroesophageal reflux. AIM: To evaluate the diagnostic accuracy of MII-pH as compared with conventional pH monitoring in detecting reflux events (REs) and symptom association in different age groups. METHODS: : A prospective direct comparison of 2 diagnostic techniques on 291 consecutive patients referred for suspected gastroesophageal reflux disease. Sensitivity and diagnostic accuracy of MII-pH versus pH monitoring and symptom association were measured. RESULTS: MII-pH detected 13631 REs, 6260 (46%) of which were nonacid. The prevalence of weakly acid refluxes in the 24 hours and postprandial period as well as the proximal extension of refluxate were significantly greater in infants as compared with children (P < 0.001, P < 0.001, and P < 0.01, respectively). The diagnostic accuracy of combined MII-pH in revealing all RE and acid RE were significantly higher in infants as compared with children (92% vs 82%, P < 0.01 and 83% vs 76%, P < 0.04, respectively). The addition of MII to conventional pH monitoring significantly increases the diagnostic yield of symptom association analysis in revealing an association between atypical symptoms and refluxes irrespective of age, whereas in studying typical symptoms it was true only for infants. CONCLUSIONS: Addition of MII to conventional pH monitoring significantly increases the diagnostic yield in detecting REs, prevalently in infants, and in revealing an association between refluxes and symptoms, prevalently respiratory ones and in infants group. PMID- 19680155 TI - Congenital and putatively acquired forms of sucrase-isomaltase deficiency in infancy: effects of sacrosidase therapy. PMID- 19680156 TI - Diagnostic use of video capsule endoscopy in a toddler with occult gastrointestinal bleeding. PMID- 19680157 TI - Simplified management of low-energy projectile extraperitoneal rectal injuries. AB - INTRODUCTION: Since minimally invasive approach for fecal diversion in low velocity extraperitoneal rectal injuries (EPRI) has a lower morbidity than open, and since computed tomography (CT) scan is helpful in ruling out concomitant intraperitoneal injuries (IPI), we utilized both modalities to simplify management of such injuries. METHODS: Retrospective review of stable patients with EPRI, treated with laparoscopic assisted diversion, after a CT scan, compared with patients with similar injuries that had a negative laparotomy and a colostomy. RESULTS: The laparoscopy group had a shorter length of stay (3 +/- 2 days versus 7 +/- 2 days), earlier return of bowel function (3 +/- 2 days versus 5 +/- 2 days) and fewer infectious complications. If the CT scan was negative for IPI, none were found on open exploration or laparoscopy. CONCLUSION: If IPI are ruled out with a preoperative CT scan, laparoscopy-assisted colostomy is safe and less morbid. PMID- 19680158 TI - Effect of polarized light emitting diode irradiation on wound healing. AB - BACKGROUND: We propose a new phototherapy using polarized light from light emitting diode (LED). The purpose of this study is to clarify the effect of polarized LED irradiation on wound healing. METHODS: Five groups were classified: control (C), unpolarized (U), linearly polarized (L), right circularly polarized (RC), and left circularly polarized (LC) LED irradiation. In vitro study, fibroblast cell cultures were irradiated, and cellular proliferation was evaluated with a WST-8 assay. In vivo study, full-thickness skin defect of 20 mm diameter was created on the dorsal side of rats. The ratio of the residual wound area was measured, and expression of type 1 and type 3 procollagen mRNA in granulation tissue was determined by real-time reverse transcription polymerase chain reaction method. RESULTS: The cellular proliferation rates of group RC and L were significantly higher than other groups. The ratio of the residual wound area of group RC and L was significantly reduced than group C and U. Expression of type 1 procollagen mRNA in group RC was found to be significantly increased about 1.5-fold in comparison with the group C. There were no significant differences for type 3 procollagen. CONCLUSIONS: The right circularly polarized light and linearly polarized light promoted the process of wound healing by increasing the proliferation of fibroblasts, and the right circularly polarized light increased the expression of type 1 procollagen mRNA. The effectiveness of right circularly polarized light suggests that some optical active material, which has a circular dichroic spectrum, takes part in a biochemical reaction. PMID- 19680159 TI - Defining impairment and treatment of subacute and chronic fractures of the coracoid process. AB - BACKGROUND: Concerning subacute and chronic coracoid fractures, the details of these disorders and its consequent treatment methods have not yet been established. METHODS: Fifteen patients who received treatment at 23 weeks +/- 23 weeks after injury were followed up for 23 months +/- 10 months. There were 10 type I fractures and 5 type II fractures according to the classification proposed by Ogawa et al. Associated shoulder girdle injuries were more frequent in the type I fractures. RESULTS: Concerning the treatment for type I fractures, all of the four patients first seen within 5 weeks after injury and adequately treated during this period were conservatively treated. The other six patients visiting later than 5 weeks required surgery because of severe motion-produced pain. Of type II fractures, four were asymptomatic at the first visit and the remaining one with atypical subcoracoid impingement symptoms needed surgery. In the type I fracture group, except one patient, all four conservatively treated and five surgically treated patients exhibited no symptoms at follow-up. All five patients with type II fractures developed nonunion, of whom four patients remained free from pain. CONCLUSION: In the overlooked and untreated type I fracture with persistent pain and functional impairment, reduction and fixation of the coracoid fracture aimed at reconstruction of the firm scapuloclavicular connection and structural restoration of the coracoacromial arch results in gratifying outcomes. In the cases of type II fracture, conservative treatment is indicated. When presenting with atypical manifestations of subcoracoid impingement, releasing of the coracoacromial ligament proves effective. PMID- 19680160 TI - Blunt cardiac rupture: a 5-year NTDB analysis. AB - OBJECTIVE: Because of its rarity and high rate of mortality, traumatic blunt cardiac rupture (BCR) has been poorly studied. The objective of this study was to use the National Trauma Data Bank to review the epidemiology and outcomes associated with traumatic BCR. METHODS: After approved by the institutional review board, the National Trauma Data Bank (version 5.0) was queried for all BCR occurring between 2000 and 2005. Demographics, clinical injury data, interventions, and outcomes were abstracted for each patient. Statistical analysis was performed using an unpaired Student's t test or Mann-Whitney U test to compare means and chi analysis to compare proportions. Stepwise logistic regression analysis was performed to identify independent predictors of inhospital mortality. RESULTS: Of 811,531 blunt trauma patients, 366 (0.045%) had a BCR of which 334 were available for analysis, with the mean age of 45 years, 65% were men, and their mean Injury Severity Score was 58 +/- 19. The most common mechanism of injury was motor vehicle collision (73%), followed by pedestrian struck by auto (16%), and falls from height (8%). Twenty-one patients (6%) died on arrival and 140 (42%) died in the emergency room. The overall mortality for patients arriving alive to hospital was 89%. Of the patients surviving to operation, 42% survived >24 hours of which 87% were discharged. Survivors were significantly younger (39 vs. 46 years, p = 0.04), had a lower Injury Severity Score (47 vs. 56, p = 0.02), higher Glasgow Coma Scale (10 vs. 6, p < 0.001), and were more likely to present with an systolic blood pressure >or=90 mm Hg (p = 0.01). Nevertheless, none of these factors was found to be an independent risk factor for mortality. CONCLUSION: BCR is an exceedingly rare injury, occurring in 1 of 2400 blunt trauma patients. In patients arriving alive to hospital, traumatic BCR is associated with a high mortality rate, however, is not uniformly fatal. PMID- 19680161 TI - Osteoporosis treatment and atrial fibrillation: alendronate versus raloxifene. AB - OBJECTIVES: Concerns have been raised about bisphosphonate use and risk of atrial fibrillation (AF) in women with osteoporosis. This study compares the risk of AF and of flutter or acute myocardial infarction (AMI) in women with osteoporosis taking alendronate or raloxifene. METHODS: Using Taiwan's National Health Insurance database to conduct a population-based retrospective cohort study, we reviewed the medical and prescription histories of 27,257 women with osteoporosis (21,037 receiving alendronate and 6,220 receiving raloxifene) between 2001 and 2007. Mean (SD) follow-up was 303.62 (422.87) days. For the main outcome measures, we calculated the adjusted relative risk of AF and AMI using the Cox proportional hazards model, adjusting for various confounders. RESULTS: Incidence rates (per patient-year) of AF in the alendronate group (1.00%) and the raloxifene group (1.02%) were similar. Alendronate use was not associated with risk of AF (hazard ratio [HR], 1.06; 95% CI, 0.85-1.32) and AMI (HR, 1.02; 95% CI, 0.86-1.19) compared with raloxifene use. However, alendronate users who had previous cardiovascular events and had taken their medications for more than 1 year were at significantly greater risk of AMI than were the group taking raloxifene (HR, 2.24; 95% CI, 1.07-4.71). Users who received 70 mg of alendronate once a week were at significantly lower risk of AF than were those taking 10 mg daily (HR, 0.56; 95% CI, 0.47-0.68). CONCLUSIONS: Compared with raloxifene, alendronate did not increase the risk of AF and flutter in women with osteoporosis. Medical history contributed most to the development of AF or AMI in the women who received either raloxifene or alendronate. Long-term treatment with alendronate is not suggested for women with a history of cardiovascular events because they are at increased risk of AMI. PMID- 19680162 TI - Growth differences by age of menarche in African American and White girls. AB - BACKGROUND: Because of the rapid increases in childhood obesity coupled with decreases in the median age of menarche, there is interest in how growth (body mass index [BMI] and height) in childhood may be associated with timing of menarche. OBJECTIVES: Two research questions were addressed in this article: (a) Within each race, at what ages were BMI and height differences evident among the early-, the mid-, and the late-onset groups? And (b) within each timing group, at what ages were BMI and height differences evident between White and African American girls? METHODS: The mother/child files of the National Longitudinal Survey of Youth were used for this study. Menarcheal timing groups were identified using the 25th and the 75th percentile of the age distribution for each race. Longitudinal statistical techniques were used to estimate BMI and height as polynomial functions of age and age relative to menarche for African American and White girls. RESULTS: Significant differences in BMI by timing group were found. By 3 years of age, significant differences were found between early- and mid-onset African American girls, by 5 years of age between mid- and late onset African American girls, and by 6 years of age among the three timing groups of White girls. Significant height differences were evident by 5 years of age when comparing early- to mid-onset and mid- to late-onset girls in both race groups. Comparing across race and within timing group, BMI and height differences were evident. African American girls were more likely than White girls to experience accelerated growth and earlier menarche. DISCUSSION: This is one of the few longitudinal studies of differences in growth by timing of menarche that includes data on girls younger than 5 years with large samples of both African American and White girls. Understanding when differences are first apparent is critical in establishing the critical period for prevention of these high-risk growth patterns. PMID- 19680163 TI - Validity and reliability of an oral care practice survey for the orally intubated adult critically ill patient. AB - BACKGROUND: Oral care is proposed as key to preventing ventilator-associated pneumonia, yet little work has been done to measure reliably current oral care practices nationwide. Five critical care oral care surveys are described in the literature; however, their usefulness is diminished because of insufficient validity or reliability measures and sampling limitations that limit generalizability. OBJECTIVES: The aim of this study was to present a survey instrument for oral care practices for the orally intubated adult critically ill patient and the attendant psychometric properties. METHODS: The oral care survey items were designed to elucidate information on the type and frequency of each specific task for oral care (toothbrushes, foam swabs, suctioning, and chlorhexidine gluconate oral rinse). The oral care survey was evaluated for face validity, content validity, and stability reliability psychometric properties. RESULTS: Face validity was established by an expert panel. Content validity was assessed using content validity index (CVI) and pairwise comparison. The overall CVI score was 97.5% agreement. A pairwise comparison for the three raters was computed. The strongest rating agreements were between Rater 1 and Rater 2 (.86) and Rater 1 and Rater 3 (.83). Given these interrater reliability scores and the overall 97.5% CVI score, minor revisions were made for survey items as recommended by the reviewers' comments. Test-retest reliability (.82-.86) showed a less than 10% difference between all items from Time 1 and Time 2. IMPLICATIONS: This survey, tested for validity and reliability, can be used in future critical care settings as an audit tool for oral care practices performed by nurses. PMID- 19680164 TI - Severe abdominal trauma involving bicycle handlebars in children. AB - OBJECTIVES: To emphasize the severity of the underlying injury which may not be realized during the initial patient admission to the emergency department. METHODS: A retrospective case note review of children admitted to our institution with the severe abdominal injury. RESULTS: Eight children were identified with the severe abdominal injury secondary to the trauma from a bicycle handlebar that needed special care in the intensive care unit. All injuries were due to blunt trauma. The mean delay from the time of the accident to the time of presentation was 34.5 hours. All patients had an imprint of the handlebar edge on the hypochondrium. There were 3 pancreatic lacerations, 1 duodenal laceration, 1 jejunal laceration, 1 liver laceration, 1 abdominoinguinal laceration that all required open surgery, and 1 duodenal hematoma that resolved in 4 weeks follow-up period. The patients who required open surgery were evaluated with computed tomographic scans before surgery. CONCLUSIONS: Children with an imprint made by the handlebar edge on the abdominal wall or give a clear history of injuries by a bicycle handlebar should be treated with great care. Early computed tomography evaluation may help to reduce the morbidity resulting from the delay in diagnosis of injuries to the internal organs. PMID- 19680166 TI - Reference values of urinary neutrophil gelatinase-associated lipocalin in very low birth weight infants. AB - In very low birth weight (VLBW) infants, acute renal impairment (ARI) is common, but there is no consensus about criteria for its diagnosis. Neutrophil gelatinase associated lipocalin (NGAL) is an early and sensitive indicator of renal impairment in experimental animals, children, and adults. Urinary NGAL (UNGAL) is detectable in VLBW infants; however, there is no reference range in this population. The objective of this study is to define the reference range for UNGAL in VLBW infants with no risk factors for acute renal impairment. UNGAL concentration was determined in urine samples collected from day of life (DOL) 4 through DOL 30 in 50 newborns with uncomplicated clinical courses, selected from a total of 145 prospectively enrolled appropriate for gestational age inborn VLBW premature infants. The birth weight and gestational age ranges were 790-1490 g and 26-33 wk, respectively. The median, 95th and 99th percentiles, and range of pooled UNGAL values were 5 ng/mL, 50 ng/mL, 120 ng/mL, and 2-150 ng/mL, respectively. Greater variability and higher quantile levels of UNGAL were observed in females versus males. In conclusion, a reference range for UNGAL in VLBW infants, similar to that in children and adults, has been established. PMID- 19680165 TI - Oxygen concentration and pulmonary hemodynamics in newborn lambs with pulmonary hypertension. AB - The effect of oxygen concentration on lowering pulmonary vascular resistance (PVR) during resuscitation in a model of persistent pulmonary hypertension of the newborn (PPHN) is not known. PPHN was induced in fetal lambs by ductal ligation 9 d before delivery. After delivery by cesarean section, resuscitation of PPHN lambs with 21%, 50%, or 100% O2 (n = 6 each) for 30 min produced similar decreases in PVR. Lambs were then ventilated with 50% O2 for 60 min and exposed to inhaled nitric oxide (iNO, 20 ppm). Initial resuscitation with 100% O2 significantly impaired the subsequent response to iNO compared with 21% O2 (42 +/ 9% vs 22 +/- 4% decrease from baseline PVR). Finally, each lamb was randomly and sequentially ventilated with 10%, 21%, 50%, or 100% O2. PVR decreased with increased concentrations of inhaled O2 up to 50%, there being no additional decrease in PVR with 100% O2. When PVR was correlated with Pao2, the maximal change in PVR was achieved at Pao2 values <60 mm Hg. We conclude that resuscitation with 100% O2 does not enhance pulmonary vasodilation compared with 21% and 50% O2, but impairs the subsequent response to iNO in PPHN lambs. Hypoxia increases PVR but hyperoxia does not confer significant additional pulmonary vasodilation in lambs with PPHN. PMID- 19680168 TI - A grandparent-influenced locus for alcohol preference on mouse chromosome 2. AB - OBJECTIVE: Loci on mouse chromosome 2 have previously been associated with ethanol consumption. Here, we used a limited access choice paradigm in which mice consume large quantities of ethanol (2-3 g/kg/2 h) with a high preference (>80%). In addition, mouse chromosome substitution strains were used to further evaluate the contribution of chromosome 2 to ethanol consumption. METHODS AND RESULTS: First, we compared the two parental inbred mouse strains, C57BL/6J and A/J, in the limited access choice paradigm for ethanol intake and ethanol preference, as well as for ethanol metabolism and taste sensitivity. Then, the effect of chromosome 2 substitution on these measures was determined. Compared with C57BL/6J mice, A/J and C57BL/6J-Chr 2/NaJ (CSS-2) mice showed profoundly reduced ethanol intake and preference. The strains were not different with regard to ethanol metabolism or taste sensitivity. Limited access ethanol consumption in F2 progeny derived from reciprocal C57BL/6J xCSS-2 and CSS-2 xC57BL/6J intercrosses and subsequent quantitative trait loci mapping identified two loci: one locus on chromosome 2 for ethanol intake and a separate locus on distal chromosome 2 for ethanol preference. This latter locus was dependent on the grandparental origin. CONCLUSION: Using a limited access choice paradigm, we found that mouse chromosome 2 carries an allelic variant of a locus for ethanol intake and a distinct locus selective for ethanol preference. The heritability of alcoholism has been suggested to be parent-specific, perhaps resulting from genetic imprinting. Our findings suggest that grandparent-influenced vulnerability for ethanol consumption is conferred by genes on chromosome 2, providing important new leads to enhance our understanding of the heritability of alcoholism. PMID- 19680167 TI - Clinical score and transcript abundance patterns identify Kawasaki disease patients who may benefit from addition of methylprednisolone. AB - Intravenous immunoglobulin (IVIG) treatment-resistant patients are high risk of developing coronary artery lesions with Kawasaki disease. The IVIG-responsive (Group A; n = 6) and IVIG-resistant patients (Group B) were predicted before starting the initial treatment using the Egami scoring system and randomly allocated as a single-IVIG treatment group (group B1; n = 6) or as a IVIG-plus methylprednisolone (IVMP) combined therapy group (group B2; n = 5). We investigated the transcript abundance in the leukocytes of those patients using a microarray analysis. Five patients in group A and one patient in group B1 responded to initial IVIG treatment. All group B2 patients responded to IVIG-plus IVMP combined therapy. Before performing these treatments, those transcripts related to IVIG resistance and to the development of coronary artery lesions, such as IL1R, IL18R, oncostatin M, suppressor of cytokine signaling-3, S100A12 protein, carcinoembryonic antigen-related cell adhesion molecule-1, matrix metallopeptidase-9, and polycythemia rubra vera-1, were more abundant in group B patients in comparison with group A patients. Moreover, those transcripts in group B2 patients were more profoundly and broadly suppressed than group B1 patients after treatment. This study elucidated the molecular mechanism of the effectiveness of IVIG-plus-IVMP combined therapy. PMID- 19680169 TI - Use of cultured human epithelium for coverage: a defect of radial forearm free flap donor site. AB - The radial forearm free flap has been popular in many areas of reconstructive surgery. Despite the many attributes of this flap in maxillofacial reconstruction, one of the disadvantages has been the morbidity of the donor site. Allogeneic cultured epidermis has been successfully applied on large second degree burns and on chronic leg ulcers. Autologous human keratinocytes and fibroblast equivalents can be cultured in-vitro from a small skin sample in order to produce a sufficient amount of epithelial autografts to cover the large defects of third-degree burn wounds. Interestingly, transplanted cultured epidermis retains characteristics of the original donor site. We report a case of a patient who underwent skin replacement by cultured epithelial autograft after wound breakdown occurred in the forearm donor site during the early postoperative period. This method could represent an auspicious alternative to conventional grafting methods for forearm free flap reconstruction. To the best of our knowledge, skin replacement by cultured epithelial autografts in this region has not been extensively described in the literature. PMID- 19680170 TI - Oral lichenoid lesions related to contact with dental materials: a literature review. AB - Oral lichenoid lesions related to contact are defined as oral-cavity eruptions with an identifiable etiology, and are clinically and histologically similar to oral lichen planus. Within this group are found oral lichenoid lesions related to contact with dental materials (OLLC), the most common being those related to silver amalgam. Currently, it remains difficult to diagnose these lesions due to the clinical and histopathological similarity with oral lichen planus and other oral mucosa lesions of lichenoid characteristics. In the present paper, we carry out an updated review of the tests for, and the different characteristics of OLLC, which may aid the diagnosis. For this review, we made searches in the Pubmed(R) and Cochrane(R) databases. Among the literature we found several published papers, from which we have used review papers, case papers, cohort studies, case and control studies, and a meta-analysis study. After carrying out this review, we can conclude that the diagnosis of these lesions is still difficult and controversial. However, there are different aspects in the clinical presentation, pathological study and results obtained when replacing suspect materials, which, when taken together, may be useful when establishing the final diagnosis of OLLC. PMID- 19680171 TI - In vitro experimental study of bonding between aluminium oxide ceramics and resin cements. AB - To evaluate bond strengths of different resin cements to two aluminum oxide-based ceramics. METHODS: One hundred ten ceramic cylinders were produced and given four different surface treatments. Resin cement cylinders were then bonded to the ceramic cylinders using different resin cements and the bond strength was determined by shear testing to the breaking point. We were thus able to obtain results for the different combinations of porcelain, surface treatments and cements. All data was analyzed using the Kruskal-Wallis test for more than two independent samples and the Bonferroni correction applied (a=0.01). An optical microscopy study was carried out to analyze the type of failure, and an electronic microscopy examination was carried out in order to evaluate the changes produced in the ceramic by the different surface treatments. RESULTS: The best values corresponded to the control group, composed of silicate ceramics combined with Variolink II resin cement. In-Ceram Alumina showed no significant differences with respect to the type of cement applied. Procera AllCeram obtained the best values when silica coated using the CoJet System and applying Variolink II, or when sandblasted and applying Clearfil SE Bond + Porcelain Bond Activator and Panavia F cement. SIGNIFICANCE: Surface treatment modifies the ceramic surface and influences the bond strength, as does the type of cement used. Silica coating is recommended to improve adhesion to Procera AllCeram, applying Variolink II, or sandblasting plus resin cement containing MDP (Panavia F). PMID- 19680172 TI - Oromandibular dystonia: a dental approach. AB - Oromandibular dystonia consists of prolonged spasms of contraction of the muscles of the mouth and jaw. Primary idiopathic forms and secondary forms exist. Secondary dystonia develops due to environmental factors; some cases of cranial dystonia after dental procedure have been reported, but the causal relationship between these procedures and dystonia remains unclear. Traumatic situations in the mouth, such as poor aligned dentures or multiple teeth extractions may cause an impairment of proprioception of the oral cavity, leading to subsequent development of dystonia. The clinical characteristics of oromandibular dystonia are classified according to the affected muscles. The muscles involved may be the muscles of mastication, muscles of facial expression, or the muscles of the tongue. At present, there is no known cure for OMD. The mainstay of treatment for most focal dystonia is botulinum toxin injections. It is important for the dentist to be familiar with oromandibular dystonia, as it can develop after dental treatment and is often misdiagnosed as a dental problem. PMID- 19680173 TI - Molecular detection of Helicobacter pylori in oral samples from patients suffering digestive pathologies. AB - OBJECTIVE: to determine the simultaneous presence of H. pylori in both the oral cavity and gastric mucosal in patients suffering digestive pathologies and to establish the possible association between the presence of H. pylori in the oral cavity and the gingivoperiodontal pathology. STUDY DESIGN: Patients with gastric symptoms (case group) and asymptomatic patients (control group) seen at the Gastroenterology Department of Dr. Julio C. Perrando Hospital (Resistencia, Argentina) were selected. Dental plaque and saliva samples from both groups were obtained. In the case group, gastric biopsy samples were also taken. H. pylori was detected in gastric biopsies by histological stains, and Polymerase Chain Reaction (PCR) was carried out on the oral samples. RESULTS: Among the 98 patients (43 cases and 55 controls), 196 oral samples (saliva 98, dental plaque 98) and 43 gastric biopsias were obtained. H. pylori was detected in oral samples in 18/98 patients, in gastric biopsies in 38/43 patients, and in both samples in 15/43 patients. CONCLUSIONS: The presence of H. pylori in the oral cavity of patients suffering digestive pathologies is more frequent in those patients harbouring a gingivoperiodontal disease, and this fact could represent an obstacle for the eradication of the bacterium. At the same time, it could constitute a risk factor for gastrointestinal reinfection after treatment. PMID- 19680174 TI - Primary de novo intraosseous carcinoma: report of a new case. AB - Primary de novo intraosseous carcinoma of the jaws has been rarely reported. We present a new case of this unusual tumour and discuss its histopathological and clinical aspects. The subject was a 76-year-old man who was seen due to complaints of pain and the presence of gingival changes in the left mandible. A panoramic radiograph and computed tomography revealed a large mandibular radiolucency. A segmentary mandibulectomy was performed and histopathologic examination proved that the tumour was an intraosseous squamous cell carcinoma. Surgeons should appreciate the aggressiveness of this tumour, despite adequate surgical treatment. PMID- 19680175 TI - Periodontal disease affecting tooth furcations. A review of the treatments available. AB - The molars are the teeth that suffer the greatest periodontal destruction in untreated patients. When periodontal disease affects the furcation of a tooth, the chance that it will be lost increases considerably. An increase in the exposed root surface, anatomical peculiarities and irregularities of the furcation surface all favor the growth of bacteria. These problems make it harder for the patient to maintain hygiene, and impede adequate treatment. The treatment of furcations affected by periodontal disease is one of the most difficult problems for the general dentist and periodontist. The motivation of both the attending professional and of the patient are therefore of great importance. No ideal procedure for treating such lesions exists. The present paper reviews those options that are currently available. Long-term research will be needed, along with the development of new techniques, to solve the problem of furcations affected by periodontal disease, possibly including substitution of the affected tooth by an implant. PMID- 19680176 TI - Hybrid salivary gland tumor of the upper lip or just an adenoid cystic carcinoma? Case report. AB - A 65 year-old male patient with a one year-duration tumoral growth located in the upper lip was diagnosed on incisional biopsy as epithelial-myoepithelial carcinoma. After wide surgical excision the histopathological analysis revealed the lesion was composed predominantly (>90%) of adenoid cystic carcinoma. In new sections it was found a very small and isolated area of adenoid cystic carcinoma at the bottom of the incisional biopsy. As surgical margins were free of lesion, no adjuvant treatment was given. The occurrence of a transitory ischaemic attack at 36 months of follow-up led to a neurological and MRI evaluation, which disclosed a well-defined 3.5 x 3 cm lesion suggestive of metastasis, located on the right temporal area. The lesion was surgically removed and a histopathological diagnosis of neurocysticercosis was rendered. After 40 months of follow-up there is no evidence of recurrence. CONCLUSION: True hybrid tumors of salivary glands are rare and treatment in each case should be done according to the component with the higher aggressiveness. However, the occurrence of epithelial-myoepithelial carcinoma areas within an adenoid cystic carcinoma seems to be a frequent finding, and because both lesions share a common origin, some authors consider that this may not be a true hybrid neoplasm but a variant of the latter. PMID- 19680177 TI - Oral health in 6-year-old schoolchildren from Berisso, Argentina: falling far short of WHO goals. AB - OBJECTIVES: Dental diseases in children have been reduced in recent decades in developed countries, although trends remain unclear in other countries. Oral healthcare in Argentina is based on demand and depends on a patient's health insurance coverage. The objective of this study was to determine the oral health situation of the population of six-year-olds in Berisso, Buenos Aires province (Argentina). STUDY DESIGN: A cross-sectional observational study was performed on schoolchildren from public and private schools. The following factors were evaluated: DMFT, DMFS, dft, dfs, significant caries (SiC), filled, decayed and missing teeth, and the prevalence of caries in both primary and permanent teeth. Data was recorded concerning malocclusion, ankylosis, dental fluorosis, DDE index modified, urgent healthcare needs, healthcare system use, orthodontic treatments, filling materials, school type and socioeconomic position. RESULTS: The study was carried out on 804 schoolchildren. The overall prevalence of caries was 70% (temporary dentition 67.9%, permanent dentition 16.3%). The dft index was 4.52 for males and 4.77 for females. For males, dfs index scored 8.78 and for females, it scored 9.27. DMFT index was 0.45 for males and 0.51 for females. DMFS index scored 0.68 for males and 0.80 for females. There were differences between socioeconomic groups (employees and manual workers) in DMFT and DMFS indexes. Of the study population, 54% had never been seen by a dentist prior to the study. For children who had visited a dentist, 71% attended state public services. CONCLUSIONS: Oral indices in Berisso were worse than in other Argentinean studies and were far from the World Health Organization global goals. There is an urgent need to strengthen the effectiveness of preventive care. PMID- 19680179 TI - HPV infection and oral carcinogenesis. PMID- 19680178 TI - Cytotoxic effects of two acid solutions and 2.5% sodium hypochlorite used in endodontic therapy. AB - AIM: To evaluate the cytotoxicity of 15% citric acid, 5% phosphoric acid and 2.5% NaOCl on cultured fibroblasts using MTT colorimetric assay. METHODOLOGY: Irrigating solutions of 5% phosphoric acid, 15% citric acid, and 2.5% NaOCl, diluted at 0.1% and 0.5%, were applied to cell cultures of 3T3L1 fibroblasts. The cell viability was determined by means of MTT colorimetric assay after a period of 1, 6 and 24 hours. Percentages of cell viability were analyzed using the Kruskal-Wallis test for global comparisons and the Mann-Whitney U-test for pairwise comparisons. RESULTS: The percentage of cell viability diminished progressively over a 24 hour period in all solutions at both dilutions. At 0.1% dilution, 2.5% NaOCl (63.39%) and 15% citric acid (53.91%) showed the highest percentage of cell viability (p=0.083). At 0.5% dilution, 2.5% NaOCl again showed the highest cell viability value (48.51%). CONCLUSIONS: The irrigating solution with the highest percentage of cell viability was 2.5% NaOCl at both 0.1% and 0.5% dilutions. A very low percentage of cell viability was obtained with 15% citric acid and 5% phosphoric acid at 0.5% dilution. PMID- 19680180 TI - Acellular dermal matrix in soft tissue reconstruction prior to bone grafting. A case report. AB - When hard tissue augmentation is scheduled as a part of an oral rehabilitation, prior to the treatment, it is important to assess if the quality of the underlying gingiva at the recipient site can support the bone grafting procedure. The most frequent complication during autologous onlay grafts are wound dehiscences in the recipient site, so the integrity of soft tissues is a basic aspect of successful reconstructive and plastic surgical procedure. Connective tissue grafts can improve the quality and quantity of soft tissue in oral sites where a hard tissue reconstruction is going to take place. However, particularly when large grafts are harvested, the autogenous donor site can present significant postoperative morbidity, such as necrosis of the palate fibromucosa and bone exposition, pain and bleeding. Another important limitation with the use of autogenous grafts is the limited supply of donor connective tissue. If a large site needs to be grafted, more than one surgical procedure may be required. An Acellular Dermal Matrix (ADM) graft has become increasingly popular as a substitute for donor connective tissue, eliminating the disadvantages described for the autogenous donor graft. The amount of tissue harvested is unlimited, so it gives an option for treating patients that have inadequate harvestable tissue or that present a large defect to be treated. The outcome of using ADM as a matrix for soft tissue reconstruction 12 weeks before bone grafting can reduce the risk of exposure and failure of the bone graft. PMID- 19680181 TI - Multiple brown tumours of tertiary hyperparathyroidism in a renal transplant recipient: a case report. AB - Tertiary Hyperparathyroidism (HPT) is an uncommon condition that affects patients with secondary HPT after a successful kidney transplant. Bone alterations are the main consequences of this endocrine condition, including the development of an osteolytic lesion called brown tumour. This paper reports an unusual case of multiple brown tumours located in the maxilla and mandible in a 58-year-old renal transplant recipient, with tertiary HPT. Incisional biopsies were performed and, in both samples, the histopathological diagnoses were giant cell lesions. Due to these diagnoses, the medical history of chronic renal failure, and high levels of serum calcium and PTH, lesions were diagnosed as multiple brown tumours of tertiary HPT. The lesions showed regression within the two months following a total parathyroidectomy. Minimal local interventions were established to make an early diagnosis and to treat these lesions. The diagnosis of primary oral lesions was very important in order to demonstrate the relevance of interdisciplinary activity and the inclusion of dentists as part of organ transplant units. PMID- 19680182 TI - Retinoids and proliferative verrucous leukoplakia (PVL). A preliminary study. AB - OBJECTIVE: A study is made of the efficacy and adverse effects of retinoid therapy applied to the white lesions of proliferative verrucous leukoplakia (PVL). MATERIAL AND METHODS: The results of retinoid therapy were evaluated in 17 patients diagnosed with PVL. Topical retinoids were used in 5 patients, in the form of two daily applications of 0.1% 13-cis-retinoic acid in orabase for an average of 6.17+/-3.13 months. Systemic retinoids were used in 11 patients, with the administration of 25 mg/day of acitretin in tablet form for an average of 5.41+/-2.02 months. One patient successively received the topical and systemic retinoid formulations. The course and results were evaluated on a blind basis by two investigators. The adverse effects of the medication were also assessed. RESULTS: Clinical improvement was recorded for 7 lesions (38.8%) (six involving systemic treatment and one as a result of topical application). Clinical worsening was recorded in the same proportion (5 lesions with systemic therapy and two with topical treatment), while four lesions (22.4%) showed no changes (one lesion with systemic therapy and three with topical treatment). Adverse effects were documented in all the patients administered the systemic formulation, versus in only one patient administered topical retinoids. The most frequent problems were desquamation and pruritus. CONCLUSION: Although topical or systemic retinoic acid produces some improvement in about one-third of all patients with PVL, further studies are needed to assess the efficacy and safety of these products, in view of the important percentage of individuals who worsen despite therapy, and the frequent appearance of adverse effects. PMID- 19680183 TI - Study of the interocclusal distortion in impressions taken with different types of closed-mouth trays and two types of impression materials. AB - The aim of this study was to compare different types of impression trays for the closed-mouth impression technique, using two different types of impression material. For this study, five different types of impression trays were used with two different types of impression materials, one of addition silicone and the other of polyether. We designed a model used for taking the impressions and for measuring interocclusal distortion. The results obtained show that the impression trays COE (GC (R) GC America INC. Alsip) and Premier (Premier (R), Premier Dental Products Co. Canada) show a lesser degree of interocclusal distortion when taking closed-mouth impressions. In terms of impression materials, the polyether was the one that produced the best results. From a clinical point of view, our study shows that the use of these types of trays is absolutely recommendable when used according to the clinical indications for which they have been designed; that said, we must not fail to consider that selecting the proper type of tray is also important. PMID- 19680184 TI - A survey of dentists' practice in the restoration of the shortened dental arch. AB - OBJECTIVES: The aim of this survey was to determine how a sample of dentists working in general dental practice in the UK sought to restore the mouths of patients who had shortened dental arches (SDAs). STUDY DESIGN: A survey was conducted over a period of six months on four commercial dental laboratories. Cases of SDAs comprising the anterior teeth and 2 to 4 premolars were examined; dental prescriptions were scrutinized and a special data collection form was completed accordingly. RESULTS: A total of 140 SDA cases were examined. Most of these cases were for mandibular SDAs (88.6%). Of the recorded SDA cases, 67.2% were restored by cobalt-chromium based removable partial dentures (RPD); 25.7% were extended by acrylic-resin based RPDs; implants were provided to restore 8 SDA cases (5.7%); and only two SDA cases (1.4%) were extended by cantilevered fixed bridges. Neither the gender of the patient ([Chi (2)= 4.19, p>0.05) nor the length of the SDA ([Chi (2) = 6.51, p>0.05) influenced the choice of prosthesis to be implemented. CONCLUSIONS: It would appear from the results of this survey that among the different restorative treatment options for SDA, the RPD was the most popular treatment option for dentists surveyed in this UK study. Extending the SDA by either an implant-supported prosthesis or cantilevered fixed bridges did not appear to be a popular choice of treatment. PMID- 19680185 TI - The frequency and characteristics of idiopathic osteosclerosis and condensing osteitis lesions in a Turkish patient population. AB - OBJECTIVES: The objective of this study was to investigate the frequency of idiopathic osteosclerosis (IO) and condensing osteitis (CO) in a Turkish patient population, considering factors such as age and sex of the population, in addition to shape and localization, as well as the dental relationship between IO and CO lesions. PATIENTS AND METHODS: A retrospective study was performed using panoramic radiographs of 6,154 patients ranging in age from 5 to 69 years old, who had been subjected to dental treatment. Descriptive characteristics of radiopacities, including shape, localization and dental relationship were recorded. The Chi-squared test was used. RESULTS: A total of 238 radiopacities were detected, which included 185 IO lesions in 150 (2.44 %) subjects (96 female, 54 male and mean age: 26.2), and 53 CO lesions in 50 (0.81 %) subjects (27 female, 23 male and mean age: 32.8). Both IO and CO lesions were found to be higher in number among females, as compared to males. However, this difference was statistically significant for IO lesions only (p<0.001). The frequency of IO lesions was found to be significantly higher in the 3rd and 4th decades of life (20-39 years) than in other decades (p<0.001). On the other hand, the frequency with which the CO lesions were detected was similar in ages ranging between 20-39 and 40-69 years old, and its frequency in these periods was noted to be statistically higher than in the 1st and 2nd decades of life (p<0.01). CONCLUSION: Our results point to the low IO and CO frequency among the Turkish population. In addition, our findings support the theory that IO lesions are developmental variations of normal bone architecture unrelated to a local stimulant, whereas CO lesions could be considered reactive formations related to teeth with severe caries, restoration or pulpitis. PMID- 19680186 TI - The atraumatic restorative treatment approach: an "atraumatic" alternative. AB - INTRODUCTION: Fear and anxiety are part of all human experiences and they may contribute directly to a patient's behavior. The Atraumatic Restorative Treatment (ART) is a technique that may be an alternative approach in treating special care patients or those who suffer fear or anxiety. OBJECTIVE: the aim of this paper is to review the ART technique as an alternative to reduce pain and fear during dental treatment. MATERIAL AND METHODS: A search for the term "atraumatic restorative treatment" was carried out in the MEDLINE search engine. References, from the last 10 years, containing at least one of the terms: "psychological aspects", "discomfort", "fear", "anxiety" or "pain", were selected. RESULTS: A total of 120 references were found, from which only 17 fit the criteria. DISCUSSION: All authors agreed that the ART promotes less discomfort for patients, contributing to a reduction of anxiety and fear during the dental treatment. Results also indicated that ART minimizes pain reported by patients. CONCLUSIONS: The ART approach can be considered as having favorable characteristics for the patient, promoting an "atraumatic" treatment. This technique may be indicated for patients who suffer from fear or anxiety towards dental treatments and whose behavior may cause the treatment to become unfeasible or even impossible altogether. PMID- 19680187 TI - Retrospective study of a series of 850 patients with temporomandibular dysfunction (TMD). Clinical and radiological findings. AB - OBJECTIVE: To define the clinical and radiological characteristics of the four most common presentations of temporomandibular dysfunction-myofascial pain (MFP), disc displacement with reduction (DDWR), disc displacement without reduction (DDWoR), and osteoarthrosis (OR)-and to identify the differences among them. MATERIAL AND METHODS: The study comprised a series of 850 patients (121 males and 729 females) seen between May 2003 and December 2006 in Valencia University General Hospital (Valencia, Spain) for temporomandibular joint disease (TMJD). An analysis was made of the possible etiological factors (stress, traumatisms, sleep disturbances, parafunctional habits, reason for consultation), possible pain sensations in response to palpation of the masticatory muscles, joint sounds, etc. A panoramic X-ray study was made on a routine basis, and in some patients (n = 54) the study was completed with a magnetic resonance imaging scan of the temporomandibular joints and related tissues. The differences between qualitative variables were examined by means of the chi-square test with R x C contingency tables and the Z-test, while quantitative variables were contrasted by analysis of variance (ANOVA) and post hoc testing (Scheffe). RESULTS: The variables showing statistically significant differences among the four diagnostic categories were: patient age, sleep disturbances, stress, parafunctional habits, nibbling on hard objects and "other parafunctions", reason for consultation, mandibular movements, "non-evaluable" molar and canine relationship, ligament hyperlaxity, and panoramic X-ray alterations. PMID- 19680189 TI - Fibronectin as an adjuvant in the diagnosis of oral inflammatory myofibroblastic tumor. AB - Inflammatory myofibroblastic tumor is a distinctive lesion composed of myofibroblastic spindle shaped cells accompanied by inflammatory infiltrate that may arise in various organs. It is believed to be a noneoplastic inflammatory condition, although this is still controversial. The recognition of inflammatory myofibroblastic tumor as an entity is important especially to avoid unnecessary surgery. A few cases have been reported in the oral cavity. This report primarily presents a case of inflammatory myofibroblastic tumor that arose in the floor of mouth of a 23-year-old woman. The proliferating spindle cells were immunoreactive for vimentin, smooth muscle actin, and muscle specific actin and negative for desmin, AE1/AE3, S-100, CD68, MyoD1 and caldesmon. In an attempt to assess the potential neoplastic nature of this lesion, immunohistochemical expression of ALK protein was performed, although no immunoreactivity was detected. Also, the presence of well differentiated myofibroblasts identified by fibronectin is discussed, as well as the importance in establishing an immunoprofile to better consolidate the diagnosis. We conclude that the study of fibronectin in case series may aid the diagnosis as well as the prediction of the tumor course. PMID- 19680188 TI - Ameloblastic fibro-odontoma: a conservative surgical approach. AB - Ameloblastic fibro-odontoma (AFO) is a rare benign mixed odontogenic tumor that occurs predominantly in children and young adults with no gender predilection and anatomic site, usually appearing as a painless swelling. We present a case of an 11-year-old non-Caucasian boy complaining of large painless isolated swelling in the right mandibular body. Intraoral examination revealed a tumoral mass with cortical bone expansion, covered by normal mucosa measuring 4.0 x 2.0 cm, located on both the lingual and buccal surfaces of the right body of the mandible, with displacement of the neighboring teeth. Panoramic radiography revealed an expansile, radiolucent and well circumscribed lesion with scattered foci of calcified material, which contained several radiopaque bodies of varying sizes and shapes. The provisional diagnoses were odontoma or AFO/ Biopsy confirmed AFO. The patient was treated with conservative surgery. After two years of follow-up, no alteration or recurrence was detected. PMID- 19680190 TI - Histomorphological study of the bone regeneration capacity of platelet-rich plasma, bone marrow and tricalcium phosphate: Experimental study on pigs. AB - INTRODUCTION: Bone defects are rather common after oral surgery and may prove difficult to repair. OBJECTIVE: We provide a histomorphological analysis of the bone regenerative capacity of platelet-rich plasma at different concentrations and the extraction of platelet-rich bone marrow, compared with beta-tricalcium phosphate. METHODOLOGY: We performed an experimental study on 8 pigs, in which we performed trepanations of the mandible in order to place the materials to be studied. Using an electron microscope, we observed the samples obtained and took a series of photographs in order to analyze the samples through a gray-scale histogram system. RESULTS: Ossification phenomena were present in 96% of the charged defects, regardless of the material used to fill it. Platelet-rich plasma (PRP) and the bone marrow (M) showed an equivalent degree of osteogenesis, 12.3 and 13.4 respectively, which is greater in than the control group. The platelet poor plasma (PPP) shows a capacity similar to the control groups (C), with an average count of 14.03 and 14.12 respectively. Tricalcium phosphate (TP) was shown to be effective as an ossification inducer, 3.03 times stronger than the control group. CONCLUSIONS: Ossification occurs in most of the charged defects. PRP and M had the greatest osteogenic capacity but PPP was no more effective than the control . PMID- 19680191 TI - Occupational stress among Brazilian oral-maxillofacial surgeons. AB - PURPOSE: The aim of the present study was to assess the stress level of oral maxillofacial surgeons, based on the Demand-Control Model. METHODS: A cross sectional study was carried out with 128 oral-maxillofacial surgeons who participated in the Brazilian Congress of Oral-Maxillofacial Surgery held in the city of Florianopolis, State of Santa Catarina, Brazil. Data was obtained using a questionnaire that incorporated the Demand-Control Model (Job Content Questionnaire), which evaluates psychosocial experiences at work. Information on age, gender and professional qualification was also collected. The data was displayed in contingency tables. The following statistical tests were used with a 5% level of significance: chi-square test and "Fisher exact test". RESULTS: Oral maxillofacial surgeons were classified according to the Demand-Control Model as follows: High Demand (28%), Low Demand (28%); Passive Work (27%), Active Work (16.4%). CONCLUSION: The majority of oral-maxillofacial surgeons exercise their profession in unsatisfactory working conditions, as less than 1/3 of the population studied worked under the ideal condition of low demand. PMID- 19680192 TI - Host defence mechanisms against bacterial aggression in periodontal disease: Basic mechanisms. AB - Periodontal diseases are complex bacteria-induced infections characterised by an inflammatory host response to plaque microbiota and their by-products. Most of these microorganisms have virulence factors capable of causing massive tissue destruction both directly, through tissue invasion and the production of harmful substances, or indirectly, by activation of host defense mechanisms, creating an inflammatory infiltrate of potent catabolic activity that can interfere with normal host defense mechanisms. In response to the aggression, host defense mechanisms activate innate and adaptive immune responses. Our aim is to offer a general overview of the main mechanisms involved in the host response to bacterial aggression in periodontitis, such as lipopolysaccharide receptor CD14, complement system, polymorphonuclear neutrophils, antibodies and immunoglobulins. PMID- 19680193 TI - Evaluation of morphological changes produced by orifice opener systems using computerized tomography (CT). AB - OBJECTIVES: To evaluate modifications in root canal morphology following instrumentation with orifice openers using Computerized Tomography (CT). STUDY DESIGN: Experimental in vitro study. A preoperative CT was performed on 49 molars to measure the root canal cross-sections. After preparing the canals with 6 different nickel-titanium (Ni-Ti) rotary system orifice opener type instruments and Gates Glidden burs, a further CT was made and compared with the preoperative one. Kruskal-Wallis and ANOVA statistical analysis were used. RESULTS: Gates Glidden burs were significantly more aggressive in the coronal portion of the canals, although all the instruments removed more dental tissue in the most coronal of the sections studied. Bucco-lingual diameters were modified more than the mesio-distal ones. The stainless steel instruments used lead to greater modifications being required in the canals than those made of nickel-titanium. The ProFile system instruments were observed to be those that caused the least amount of change. CONCLUSIONS: Stainless steel burs are more aggressive than new orifice openers without significant differences observed between Ni-Ti instruments. PMID- 19680194 TI - In vitro evaluation of the temperature increment at the external root surface after Er,Cr:YSGG laser irradiation of the root canal. AB - OBJECTIVES: A study was made to determine the temperature increment at the dental root surface following Er,Cr:YSGG laser irradiation of the root canal. DESIGN: Human canines and incisors previously instrumented to K file number ISO 30 were used. Irradiation was carried out with glass fiber endodontic tips measuring 200 mm in diameter and especially designed for insertion in the root canal. The teeth were irradiated at 1 and 2 W for 30 seconds, without water spraying or air, and applying a continuous circular movement (approximately 2 mm/sec.) in the apico coronal direction. RESULTS: At the 1 W power setting, the mean temperature increment was 3.84 degrees C versus 5.01 degrees C at 2 W. In all cases the difference in mean value obtained after irradiation versus the mean baseline temperature proved statistically significant (p<0.05). CONCLUSIONS: Application of the Er,Cr:YSGG laser gives rise to a statistically significant temperature increment at the external root surface, though this increment is probably clinically irrelevant, since it would appear to damage the tissues (periodontal ligament and alveolar bone) in proximity to the treated tooth. PMID- 19680195 TI - Costochondral graft with green-stick fracture used in reconstruction of the mandibular condyle: experience in 13 clinical cases. AB - Since its publication in 1920 by Gillies, costochondral grafts have been used by surgeons to replace and injured mandibular condyle and to reconstruct the temporomandibular joint. This procedure is currently applied in cases of congenital dysplasia, developmental defects, temporomandibular ankylosis, neoplastic disease, osteoarthritis and post-traumatic dysfunction. Over the years, various procedures for the reconstruction with this type of graft have been described. In 1989, Mosby and Hiatt described a technique for setting the graft securely, reducing the space between the graft and the mandibular area. In 1998, Monje and Martin-Granizo developed a variation of this method, enabling a precise adaptation of the costochondral graft to the remaining mandibular ramus. The aim of this study is to evaluate the functional and anatomic results of the costochondral graft treatment by green-stick fracture for reconstruction of the TMJ in the 10 years following the description of this technique. We carry out a retrospective study of thirteen cases of temporomandibular pathology (tumors, ankylosis and hypoplasia) treated during a period of ten years from 1998 to 2008. In all these cases, the technique described by Monje and Martin-Granizo was used: removal of the sixth rib, fixation to a titanium mini-plate using screws, making an internal corticotomy in order to obtain a green-stick fracture of the outer cortex, providing adequate adaptation of the graft to the mandibular ramus. The graft was then set in place, attaching it with titanium screws. This technique was successful in achieving optimal ossification, a good interincisal opening and satisfactory cosmetic results. In conclusion, according to our experience, the green-stick fracture for the adaptation of costochondral grafts to the remaining mandibular ramus has presented outstanding results in the surgical treatment of temporomandibular pathology. PMID- 19680196 TI - Ameloblastic fibro-odontoma: a case report. AB - Ameloblastic fibro-odontoma (AFO) is a rare, benign, slow-growing odontogenic tumor, generally asymptomatic and more prevalent in children and adolescents. We report a case of AFO in the mandible of an eight-year-old Caucasian male patient, and review the literature . Intraoral examination revealed a swelling extending from the deciduous second molar to the retromolar triangle, covered with normal mucosa. A panoramic radiograph showed a large, well-demarcated radiolucency with radiopaque areas. The provisional diagnosis was of AFO, and so an incisional biopsy was performed. Histologically, the lesion was composed of connective tissue resembling the dental papilla, with epithelial strands or islands, as well as denticles and amorphous masses of enamel and dentin consistent with a diagnosis of AFO. Surgical excision and curettage of the lesion were performed. The patient has been monitored for eight years and the lesion has not recurred. PMID- 19680197 TI - Oral Allergy Syndrome (OAS). General and stomatological aspects. AB - Oral Allergy Syndrome (OAS) is a special contact allergy conditioned by specific amino acids in food allergens, usually fruits, as well as some proteins from different pollens. It is usually manifested in the mouth, however relatively little is known about the syndrome in Odontology. OAS is a fairly common adverse reaction that occurs after ingesting certain foods, such as peaches or apples, produced in atopic individuals who usually suffer from rhinitis, bronchial asthma or both. This syndrome mainly affects the oral cavity and oropharynx, provoking minor local alterations, although there may be serious systematic reactions, making it important to distinguish it from other oral processes when making the diagnosis. One of the more severe forms of OAS is anaphylaxis, which is food induced. In this study, we will analyze the key etiopathogenic, clinical and therapeutic aspects of this syndrome, focusing specifically on the stomatological aspects. PMID- 19680198 TI - Comparative analysis of cell proliferation ratio in oral lichen planus, epithelial dysplasia and oral squamous cell carcinoma. AB - BACKGROUND: Although oral lichen planus has been classified by the World Health Organization (WHO) as a potentially malignant disorder, such classification is still the target of much controversy. AIM: To evaluate the cell proliferation rate in oral lichen planus, comparing it to the rate observed in epithelial dysplasia and oral squamous cell carcinoma, aiming at indications which might indicate the potential for malignant transformation. MATERIAL AND METHODS: Twenty four cases of each lesion were submitted to the streptoavidin-biotin and AgNOR technique to evaluate the immunohistochemical expression of PCNA and the mean NORs/nucleus, respectively. RESULTS: Positivity for PCNA was observed in 58.33% of oral lichen planus cases, 83.33% of epithelial dysplasia cases and 91.67% of oral squamous cell carcinoma cases. Chi-squared test showed that the number of positive cases for PCNA was significantly lower in oral lichen planus than in oral squamous cell carcinoma (p<0.05). No significant statistical difference between oral lichen planus and epithelial dysplasia (p>0.05) and between the epithelial dysplasia and oral squamous cell carcinoma (p>0.05) was observed. The mean NORs/nucleus in oral lichen planus, epithelial dysplasia and oral squamous cell carcinoma were 1.74+/-0.32, 2.42+/-0.62 e 2.41+/-0.61, respectively. Variance analysis (ANOVA) revealed significant statistical difference between oral lichen planus and the other studied lesions (p<0.05). CONCLUSION: Oral lichen planus cell proliferation rate was less than in oral epithelial dysplasia and oral squamous cell carcinoma which might explain the lower malignant transformation rate. PMID- 19680199 TI - Angiogenesis of oral lichen planus: a possible pathogenetic mechanism. AB - OBJECTIVE: Oral Lichen Planus (OLP) is a chronic inflammatory disease with an autoimmune inflammatory pathogenesis. The aim of the research is to compare the vascular endothelial growth factor (VEGF) and adhesion of molecules in the biopsy samples of patients affected by OLP, in order to research the presence of the angiogenetic phenomenon and to understand its pathogenetic mechanism. MATERIALS AND METHODS: Thirty OLP patients and thirty healthy subjects were enrolled in a study. The immunohistochemical analysis of the VEGF and vascular-endothelial adhesion molecules was carried out by means of primary antibodies and anti-CD34, anti-VEGF, anti-CD106 antigen (VCAM-1) and anti-CD54 antigen (ICAM-1). The statistical significance of the differences was checked with the Mann-Whitney test (MW test). The level of significance was set to P<0.001. Data analysis was carried out with StatView 5.0.1 (SAS Institute Inc., Cary, NC). RESULTS: The results reveal the presence of a significant angiogenesis in OLP patients for the VEGF, CD34, CD106 and CD54 (P < 0.001).. The number of vessels in the biopsies of the patients with OLP (mean +/-SD: 21.27+/-4.85), compared with the healthy subjects (mean +/-SD: 4.74+/-0.97) was significantly more (Mann-Whitney test, P < 0.001). The positive expression rate of VEGF, CD34, VCAM-1 and ICAM-1 in oral lichen samples was 64.2%, 54.3%, 32.5% and 29.7%, respectively. Isolated endothelial cells and newly-formed micro-vessels and endothelial cells with high immune-positivity to the antibodies anti-ICAM-1 and anti-VCAM-1 were observed. CONCLUSIONS: The results of our immunohistochemical research show that a significant neoangiogenesis occurs in oral lichen planus. PMID- 19680200 TI - Dental extractions in patients on antiplatelet therapy. A study conducted by the Oral Health Department of the Navarre Health Service (Spain). AB - OBJECTIVES: Antiplatelet drugs are used to treat and prevent a wide range of cardiovascular pathologies and/or cerebrovascular accidents. Although the use of anticoagulants in dental extractions is highly protocolized, a clear control method has not yet been established for antiplatelet drugs. This study is directed at evaluating the clinical consequences of extractions in patients on antiplatelet therapy. STUDY DESIGN: The Oral Health Department of the Navarre Health Service-Osasunbidea conducted a trial on 155 patients who underwent dental extractions and were receiving antiplatelet therapy. The patients were not requested to interrupt the medication and local measures were taken to control potential haemorrhage. RESULTS: No major haemorrhages were reported. One patient had a moderate haemorrhage that required emergency care. In the remaining patients the bleeding was controlled with local measures. With regard to subsequent bleeding, no differences were observed between the various antiplatelet drugs used. The only statistically significant relationship found was between bleeding and the number of teeth extracted. CONCLUSIONS: It can be concluded that no more than 3 teeth should be removed at any one time, and for multiple extractions, the teeth should be adjacent to each other. PMID- 19680201 TI - Peripheral developing odontoma in newborn. Report of two cases and literature review. AB - Extra-osseous odontogenic tumors are rarely observed. However, it is widely accepted that the remains of odontogenic epithelium entrapped in the oral soft tissues may be a possible source for peripheral odontogenic tumors differentiation. Peripheral developing odontoma is considered exceptionally rare, since few similar cases are described in the English-related literature under diverse nomenclature, such as irregular eruption, ectopic tooth, ectopic soft tissue mesiodens, ectopic odontoma and extra-osseous tooth germ. Previously reported cases invariably affected children and surgical exploration revealed tooth germs exclusively embedded in the soft tissue without bone involvement. Microscopically, all these cases exhibited developing tooth germs composed of ameloblasts, enamel matrix, odontoblastic layer, dentin and dental papilla and the morphological findings seem to depend on the developmental stage of each tooth germ at discovery. Thus, we believe that it is relevant to report two additional cases that were recently diagnosed in Brazil and Guatemala, focusing on their nomenclature, correct diagnosis and further treatment. PMID- 19680202 TI - Results of a major ambulatory oral surgery program using general inhalational anesthesia on disabled patients. AB - OBJECTIVES: To assess the demographic characteristics and comorbidities of the group to be studied, as well as various quality indicators of a Major Ambulatory Surgery (MAS) program. Quantification of the surgical-anesthetic incidents. STUDY DESIGN: We aimed to perform a retrospective and descriptive analysis of disabled patients who had received oral ambulatory surgery under general anesthesia. Data obtained from the clinical history and telephone interview included the demographic characteristics, socioeconomic status, previous dental history, cause of the mental disability, degree of mental retardation, comorbidity measured according to the scale of the American Society of Anesthesiologists (ASA), anesthesia or preoperative surgical treatments, level of analgesia, length of stay, incidents in the Resuscitation Ward, the rate of substitution, suspensions, patients admitted, complications and the degree of patient satisfaction. RESULTS: We included 112 oral surgery procedures performed on disabled patients who were treated under general inhalational anesthesia as part of MAS during the years 2006-2007. During this period, 577 restorations, 413 extractions, 179 sealants, 102 pulpectomies, 22 root canal treatments, 17 gingivectomies and 3 frenectomies were performed. A total of 75% (78 cases) of the patients had coexisting medical pathology. The average surgery time per patient was 72.69 +/- 29.78 minutes. The rate of replacement was 100%. The rate of suspension was 1.92%. The percentage of patients readmitted was 1.92%,due to significant bleeding in the mouth, which did not require treatment and the patients were discharged from hospital 24 hours after being admitted. The rate of patients who required re-hospitalization was 3.84%. CONCLUSIONS: The MAS performed in this group, despite being on patients with high comorbidity resulted in only a low number of medical incidents reported. PMID- 19680203 TI - Metastasis of renal clear-cell carcinoma to the oral mucosa, an atypical location. AB - The majority of cases of metastatic tumors involve the mandible and some the maxilla but they are considerably less common in intraoral soft tissues. In addition, the primary tumor is known in the majority of cases; although in one third of such cases, metastasis is the first clinical manifestation. The most common primary tumors metastasizing to the mouth are lung carcinoma in men and breast carcinoma in women. An oral metastasis implies a serious prognosis, as in the majority of patients there is multiple organ involvement at the time of diagnosis. We present the case of a 52-year old patient with renal pathology who came to the emergency room due to a rapidly increasing gingival tumor. With the provisional clinical diagnosis of a pyogenic granuloma,the tumor was excised. Subsequent anatomopathological analysis revealed a tumor metastasis compatible with clear-cell carcinoma, and its renal origin was confirmed by means of immunohistochemical techniques. PMID- 19680204 TI - Papillary cystoadenoma lymphomatosum (Warthin-like) of minor salivary glands. AB - Papillary cystadenoma lymphomatosum is a benign salivary gland tumor most frequently located in the parotid gland (Warthin's tumor). Its presentation in other major, or in minor, salivary glands is rare. Clinically, it manifests as a slow growing tumor, fluctuant on palpation due to its cystic morphology. The treatment of choice is complete excision with wide tumor-free margins. We present a 73-year-old female patient with an asymptomatic tumor of 8 years evolution in the right posterior area of the hard palate. We performed surgical excision and a biopsy, which was reported as papillary cystadenoma lymphomatosum. During the post-operative examination carried out after 3 weeks, it was observed that the lesion had recurred. The lesion was re-operated, performing the excision with CO2 laser and including the periosteum to ensure complete resection of the tumor. At 10 months follow-up, there was no recurrence of the lesion. This article includes a review of this condition and discusses its most important clinical and pathologic features and therapeutic approaches. PMID- 19680205 TI - The relationship of amelogenesis imperfecta and nephrocalcinosis syndrome. AB - AIM: To analyze the prevalence and associated oral findings of nephrocalcinosis in a group of patients affected with amelogenesis imperfecta (AI). The relationship between types of AI and nephrocalcinosis were also evaluated. DESIGN: This study examines patients who were referred to Pediatric Dentistry Department of SDU between the years of 2002-2007 and who, upon clinical and radiological examination, were diagnosed with AI and treated. Patients were offered information about the possibility of nephrocalcinosis syndrome. Patients who agreed to have tests carried out on their renal system were advised to visit the department of nephrology at the clinic. RESULTS: Suspicious radiopacity was observed during renal ultrasonography of a controlled number of patients with hypoplastic type AI. Laboratory results revealed low Ca values (100-300 mg/days) and normal P values (0.4-1.3 g/days). Delayed eruption, gingival hyperplasia, pulp stones and orthodontic problems were also observed in the same patient groups. CONCLUSION: Although renal findings were observed in a few patients, pediatric dentists are the doctors who are the first to have early contact with this patient group. Because of the potential risk of nephrocalcinosis, early diagnosis may offer good prognosis. PMID- 19680206 TI - Treatment of chronic mandibular dislocations by eminectomy: follow-up of 10 cases and literature review. AB - INTRODUCTION: Temporomandibular joint (TMJ) dislocation is defined as an excessive forward movement of the condyle beyond the articular eminence, with complete separation of the articular surfaces and fixation in that position. OBJECTIVES: To report ten cases treated by eminectomy for chronic mandibular dislocations, to evaluate the results of these surgeries and make a critical review of the literature. METHODS: The sample was obtained from the records of the Department of Oral and Maxillofacial Surgery, University of Pernambuco and comprises cases submitted to chronic mandibular dislocation treatment by eminectomy between 2002 and 2007. Pre- and postoperative assessment included a thorough history and physical examination to determine the maximal mouth opening, presence of pain and sounds, frequency of luxations, recurrence rate and presence of facial nerve paralysis. RESULTS: The mean maximal mouth opening in the preoperative period was 48.4 +/- 8.5 mm and in the postoperative period it was 41.3 +/- 5.0 mm. No facial nerve paralysis or recurrence was observed. CONCLUSION: The treatment of chronic mandibular dislocations by eminectomy was shown to be efficient in relationship to the postoperative maximal mouth opening, recurrence and articular function. PMID- 19680207 TI - Correlation between oral health-related quality of life (OHQoL) and oral disorders in a Turkish patient population. AB - OBJECTIVES: The purpose of the present study is to determine the nature of the complaints that bring patients to our clinic and to what degree these complaints affect their quality of life (QoL). We also aimed to determine any associations between gender, education or harmful habits and each patient's oral health related quality of life (OHQoL). METHODS: A total of 1090 patients, consisting of 651 females (59.7 %) and 439 males (40.3 %), were included in this study. Of these patients, 220 constituted healthy controls. Two patient-centered outcome measures, the 14 item OHIP-14 and the 16 item OHQoL-UK measures were used. RESULTS: Most of the patients presented with toothache and caries (50.1 %), 11.2 % had suffered tooth loss and had denture needs, 9.2 % had periodontal problems, 1.8 % had temporomandibular joint (TMJ) disorders, 3.8 % had buried third molars, 2.4 % had orthodontic and aesthetic disorders, 1.3 % had suffered injury due to trauma, and 20.2 % came only for control checkups. OHQoL was best in the control group and the worst in patients who had suffered trauma. In addition, we noted correlations between gender, education and harmful habits, and that of the patient's oral health-related quality of life. CONCLUSION: According to our results, OHQoL is associated with the oral complaints of patients. Furthermore, OHQoL may not only be associated with the oral health status of patients, but factors such as gender, education and harmful habits may also play a role. PMID- 19680208 TI - Problem-based learning versus lectures: comparison of academic results and time devoted by teachers in a course on Dentistry in Special Patients. AB - OBJECTIVES: Problem-Based Learning (PBL) is a teaching-learning technique centred on the complete development of the student. It has been successfully implemented in several universities, notably in the health sciences. The process of creating the European Higher Education Area, initiated in Bologna, encourages European universities to use new methodologies in the teaching-learning process, including PBL. Our objectives were to analyze the results obtained by using PBL with fifth year Dentistry students. STUDY DESIGN: Comparison of academic results between students receiving lectures and PBL participants, and assessment of differences between them in the time devoted to tasks by students and teachers. RESULTS: PBL participants obtained higher grades compared to those receiving lectures only (p<0.05). The two student groups devoted the same amount of time to this subject but the time distribution of tasks was very different, with PBL students spending more time on group work and analysis of the literature. The teachers devoted much more time to the PBL group. CONCLUSIONS: PBL is a teaching-learning methodology that improves student academic results. PBL diverts student time to more complex tasks but requires a greater commitment from the teachers. PMID- 19680209 TI - Correlation of serum and salivary CA15-3 levels in patients with breast cancer. AB - OBJECTIVES: The aim of this study was to assess the relationship between serum and saliva levels of cancer antigen (CA) 15-3 and to compare them between women with and without breast cancer. STUDY DESIGN: A case-control study was carried out on 61 women aged 28-69 years, including women with and without breast cancer (26 as part of the case study and 35 as part of the control group) conducted at the Emam Khomeini Hospital, Tehran University of Medical Sciences. CA 15-3 levels were assayed in serum and unstimulated whole saliva by EIA. Unpaired t-test, one way ANOVA and Pearson correlation were used as statistical analysis. RESULTS: The salivary and serological levels of CA 15-3 in the cancer patients were significantly higher (P <0.01) than the salivary and serum levels of healthy controls. They were also higher in stage 2 than in stage 1 in cancer patients. However, the saliva flow rate was significantly lower in the cancer patients (P <0.01). There was a significant positive correlation between serum and saliva CA 15-3 concentration (r = 0.614) and also between serum concentration and saliva output of CA 15-3 (r = 0.541). CONCLUSION: The results of the study suggest that salivary CA 15-3 may have potential use in the initial detection of breast cancer in women. PMID- 19680210 TI - Human papillomavirus in saliva of patients with oral squamous cell carcinoma. AB - OBJECTIVE: The aim of this study was to evaluate the presence of human papillomavirus (HPV) in saliva rinses of patients with oral squamous cell carcinoma and to analyze the possibility of using saliva as a diagnostic method for screening high-risk patients. STUDY DESIGN: The saliva sample of 22 patients with oral squamous cell carcinoma and 20 age-sex matched healthy controls were obtained. The presence of HPV 6, 11, 16, 18, 31, and 33 was evaluated by polymerase chain reaction (PCR). RESULTS: In 40.9% of the patients and in 25% of the controls, the saliva was shown to be positive for HPV. In 27.3% of the patients and in 20% of the controls, the saliva was shown to be positive for HPV16; and none of the controls, except one patient was shown to be positive for HPV 18. Neither patients nor controls were positive for HPV 31 and 33. These differences were not statistically significant. CONCLUSIONS: The results of this study were unable to support the detection of HPV in saliva rinses as a diagnostic method for OSCC. PMID- 19680211 TI - Measurement of dental implant stability by resonance frequency analysis: a review of the literature. AB - Dental implant treatment is an excellent option for prosthetic restoration that is associated with high success rates. Implant stability is essential for a good outcome. The clinical assessment of osseointegration is based on mechanical stability rather than histological criteria, considering primary stability (absence of mobility in bone bed after implant insertion) and secondary stability (bone formation and remodelling at implant-bone interface). The aim of this study was to review the literature on Resonance Frequency Analysis (RFA) as a method for measuring dental implant stability. An online search of various databases was conducted on experimental and clinical research published between 1996 and 2008. The studies reviewed demonstrate the usefulness of RFA as a non-invasive method to assess implant stability. Further research is required to determine whether this system is also capable of measuring the degree of dental implant osseointegration. PMID- 19680212 TI - Analysis of the antibiotic prophylaxis prescribed by Spanish Oral Surgeons. AB - AIM: To identify prophylactic antibiotic prescription practices among Spanish dentists with preferential dedication to Oral Surgery in different types of tooth extraction surgeries. METHOD: Members of the Spanish Oral Surgery Society were surveyed on antibiotic prophylaxis use in 4 different tooth extraction modalities scaled according to their surgical invasiveness. RESULTS: Sixty-nine of the 105 distributed questionnaires were returned completed. Thirteen percent of the surveyed surgeons would prescribe antibiotics to prevent postoperative wound infection when confronted with conventional tooth extraction lasting less than 5 minutes. In the case of surgery lasting more than 5 minutes, the percentage of participants that would prescribe antibiotics increased to 39%. When a mucoperiosteal flap was elevated or an ostectomy was performed, 87% and 100%, respectively, would prescribe antibiotic prophylaxis. Amoxicillin and its combination with clavulanic acid were the most commonly prescribed antibiotics. All participants would prescribe the antibiotic orally, starting after surgery and with a duration that ranged from 2-8 days. CONCLUSIONS: The results obtained suggest that antibiotic prophylaxis for preventing local odontogenic infection is not being correctly implemented in Spain. This can generate new bacterial resistances, facilitate adverse drug reactions and favor opportunistic infections. Better designed studies are needed in order to clarify the role of antibiotics in the prevention of postsurgical wound infection. PMID- 19680213 TI - Incidence of mandibular fractures in Eastern part of Libya. AB - The objective of this retrospective study is to evaluate the incidence of mandibular fractures in the eastern part of Libya and to present our experience in treating this type of facial fracture. We analyzed factors such as the incidence of age, sex, time distribution, cause and site of the fracture and the associated injuries in 493 patients presenting a total of 666 mandibular fractures. These patients were treated at Al-Jala Trauma Hospital, Benghazi-Libya between 2000 and 2006. The results were obtained from 432 males and 61 females, for which the ages ranged from 8 months to 72 years. The maximum number of the patients was recorded in 2004, and the busiest month was May. The most common cause of fracture was road traffic accidents and the most common site was the parasymphysis. Among those treated with closed reduction were 241 patients, whereas 201 patients were treated with open reduction. In conclusion, we found that the results were similar to most studies from developing countries and were in contrast to other studies. This may be due factors such as geography, socioeconomic trends, religion, road traffic legislation and seasons, which differ from one country to another. The period during which there was an embargo in Libya also appears to have affected the results. PMID- 19680214 TI - Fluoride - an adjunctive therapeutic agent for periodontal disease? Evidence from a cross-sectional study. AB - OBJECTIVE: To assess the influence of the water fluoride level on periodontal status, by determining the periodontal health status of subjects residing in low, optimum and high fluoride areas. STUDY DESIGN: A cross sectional survey was carried out on 967 adults aged 35-44 years old, from the Udaipur district of India. A stratified cluster random sampling technique was implemented in order to collect a representative sample from low (<0.6 ppm), optimum (0.6 - 1.2 ppm) and high fluoride (>3 ppm) areas, based on the fluoride concentration in drinking water. Periodontal status was assessed in accordance to WHO criteria. The Chi square test was used to compare proportions, and logistic regression analysis was used to determine the contribution of water fluoride levels to periodontal disease. RESULTS: Those residing in areas of low fluoride levels were more likely to present periodontal pockets than those living in high fluoride areas 1.3 (95 % CI 1.11+/-1.86). Subjects living in areas of low fluoride were noted to have a higher risk of periodontal attachment loss of more than 8mm (OR = 1.94, 95% CI 1.67+/-3.85). The risk for presence of periodontal pockets and attachment loss of more than 8mm increased by 1.17 (95 % CI 1.02+/-1.69) and 1.59 (95 % CI 1.27+/ 3.29) respectively for those residing in areas of optimum fluoride levels. Deep periodontal pockets were more prevalent (6.3%) among those residing in areas of low fluoride, followed by optimum (5.2%) and high (3.1%). CONCLUSIONS: As the fluoride concentrations increased, the prevalence of shallow and deep periodontal pockets decreased. The severity of periodontal disease was significantly associated with fluoride levels, with cases of loss of attachment gradually decreasing when moving from low fluoride areas to high fluoride areas. It appears that longitudinal studies need to be conducted in order to ascertain the benefits; and microbiological analysis of dental plaque and periodontium should be carried out in order to confirm the effects of fluoride on periodontal conditions. PMID- 19680215 TI - Determination of salivary glucose in healthy adults. AB - OBJECTIVES: Our aim in this study was to determine the concentration of salivary glucose in healthy individuals and to compare it with the capillary glycemia. STUDY DESIGN: Samples of unstimulated whole saliva were collected from 63 non diabetic patients. The concentration of salivary glucose and capillary blood was measured in all of the patients. The salivary glucose was determined by enzymatic method and spectrophotometry. The data was then analyzed using the Spearman correlation test, considering values of p<0.05 to be significant. RESULTS: The whole sample consisted of 47.6% males and 52.4% women, with an average age of 37.5+/-15.7 years old. The average rates of unstimulated salivary flow were 0.41+/-0.21 ml/min among males and 0.31+/-0.15 ml/min among females. No significant difference was found based on these results (p=0.078). The average blood glucose among the males studied was 100.05+/-13.51 mg/dL, and among females, it was 99.5+/-13.9 mg/dL. The average salivary glucose for the whole sample was 5.97+/-1.87 mg/dL, with 5.91+/-2.19 mg/dL among males and 5.97+/-1.56 mg/dL among females, respectively, without presenting any significant differences (p=0.908). The concentration of salivary glucose did not present any statistically significant correlation with the capillary glycemia (p=0.732). CONCLUSIONS: The results suggest that the concentration of salivary glucose is not dependent on capillary glycemia and that the concentration of salivary glucose does not present significant differences between the measurements for males and females. PMID- 19680216 TI - Intravascular papillary endothelial hyperplasia: report of 4 cases with immunohistochemical findings. AB - Intravascular papillary endothelial hyperplasia (IPEH) is a benign endothelial proliferation, usually intravascular, that may mimic angiosarcoma. In this report, four new cases of IPEH involving the oral region are described. The affected sites were the lower lip, labial comissure and the submandibular region. After clinical evaluation, the complete removal of the lesions showed a circumscribed and soft mass. Histologically, the major feature was a reactive proliferation of endothelial cells composed of small papillary structures with hypocellular and hyalinized cores arising in an organized thrombus. Immunohistochemical staining for CD34 was strongly positive in endothelial cells. Vimentin and laminin immunolabelling were also consistent with a vascular origin. In order to verify the proliferative potential of the lesions, the Ki-67 antibody was used, revealing low percentage of labeled cells (<20%). No immunoreactivity for GLUT-1 was observed. Since the complete removal is curative, no additional treatment was necessary, and no signs of recurrence had been observed until now. Due to the particular features of IPEH, it is important for pathologists and clinicians to become familiar with this lesion. Additionally, the specific histological arrangement, including the absence of cellular pleomorphism, mitotic activity and necrosis, represents a guide to help in the differential diagnosis. Moreover, the vascular origin and the proliferative index should be assessed by immunohistochemistry in order to provide an accurate diagnosis. PMID- 19680217 TI - Changes in radiological protection and quality control in Spanish dental installations: 1996-2003. AB - INTRODUCTION: The European Union has established specific directives concerning radiological protection which are obligatory for member States. In addition, all Spanish dental clinics with radiological equipment are required to have an annual quality control check. OBJECTIVE: To analyze the effect of new European legislation on dental radiological practice in Spain and to determine whether it has resulted in lower doses being administered to patients. MATERIAL AND METHODS: A total of 10,171 official radiological quality control reports on Spanish dental clinics, covering 16 autonomous regions, were studied following the passing of Royal Decree 2071/1995 on quality criteria in radiodiagnostic installations. The reports, compiled by U.T.P.R Asigma S.A., a company authorised by the Nuclear Safety Council, cover the years 1996 to 2003, which has enabled us to monitor the evolution of radiological procedures in dental clinics over a seven year period. RESULTS: According to the reports for 2003, 77.3 % of clinics complied with EU requirements, using equipment of 70 kVp, 8 mA, 1.5 mm Al filters, with a collimator length of 20 cm. However, non-compliance was detected in approximately a third (30.8%) of the equipment inspected: alterations in the kilovoltage used, exposure time, performance of the tubing, dosage, linearity/intensity of current and acoustic-luminous signal 6.86%. The mean skin dose reached 3.11 mGy for patients who received an x-ray of an upper molar, representing a decrease of 18% over the seven years studied. CONCLUSION: there has obviously been a general improvement in the parameters studied, but only 77.3% of the installations complied fully with official EU regulations concerning dental radiological protection. PMID- 19680218 TI - Myofascial pain syndrome associated with trigger points: a literature review. (I): Epidemiology, clinical treatment and etiopathogeny. AB - Over the last few decades, advances have been made in the understanding of myofascial pain syndrome epidemiology, clinical characteristics and aetiopathogenesis, but many unknowns remain. An integrated hypothesis has provided a greater understanding of the physiopathology of trigger points, which may allow the development of new diagnostic, and above all, therapeutic methods, as well as the establishment of prevention policies and protocols by the health profession. Nevertheless, randomized studies are needed to provide a better understanding and detection of the different factors involved in the origin of trigger points. PMID- 19680219 TI - Carbon monoxide exposures after hurricane Ike - Texas, September 2008. AB - During power outages after hurricanes, survivors can be at risk for carbon monoxide (CO) poisoning if they use portable generators improperly. On September 13, 2008, Hurricane Ike struck the coast of Texas, leaving approximately 2.3 million households in the southeastern portion of the state without electricity. Six days later, 1.3 million homes were still without electrical power. To assess the impact of storm-related CO exposures and to enhance prevention efforts, CDC analyzed data from five disparate surveillance sources on CO exposures reported during September 13--26 in counties of southeast Texas that were declared disaster areas by the federal government. This report describes the results of that analysis, which indicated that one data source, Texas poison centers, received reports of 54 persons with storm-related CO exposures during the surveillance period. Another data source, the Undersea and Hyperbaric Medical Society (UHMS) hyperbaric oxygen treatment database, reported that 15 persons received hyperbaric oxygen treatment for storm-related CO poisoning. Medical examiners, public health officials, and hospitals in Texas reported that seven persons died from storm-related CO poisoning. Among the data sources, the percentage of reported storm-related CO exposures caused by improper generator use ranged from 82% to 87%. These findings underscore the need for effective prevention messages during storm preparation, warnings, and response periods regarding the correct use of generators and the installation and maintenance of battery-powered CO detectors. PMID- 19680220 TI - Pseudo-outbreak of legionnaires disease among patients undergoing bronchoscopy - Arizona, 2008. AB - Legionnaires disease (LD) is a potentially fatal form of pneumonia acquired by inhalation of aerosolized water containing Legionella bacteria. Legionella is a common cause of health-care--associated pneumonia, particularly in settings with hematopoietic stem-cell or solid-organ transplant recipients. On July 25, 2008, the Arizona Department of Health Services (ADHS) notified CDC of four patients who had Legionella cultured from specimens obtained during bronchoscopies performed at a medical center in Arizona. To characterize transmission and identify the source, ADHS and CDC began an investigation on August 1. This report summarizes the results of that investigation, which determined that the patients did not have LD and that nonsterile ice used to cool saline-filled syringes for bronchoalveolar lavage was the likely source of Legionella contamination of these clinical specimens. Ice was supplied by two ice machines, which became contaminated by heavy Legionella colonization within the center's potable water supply during a 6-month period (February--July 2008). Findings from the investigation underscore the importance of adherence to recommended infection control practices and surveillance for LD in health-care settings. Clinicians and endoscopy technicians should ensure that nonsterile items are not introduced during bronchoscopy procedures. PMID- 19680221 TI - Hepatitis temporally associated with an herbal supplement containing artemisinin Washington, 2008. AB - Artemisinins are a class of compounds that include artesunate, artemether, and artemisinin and have potent antimalarial activity. In combination with other drugs (artemisinin combination therapy), these compounds are the first-line treatment recommended by the World Health Organization for Plasmodium falciparum infections. Artemisinins have been available in the United States without a prescription as herbal supplements for at least 10 years; these supplements are marketed for general health maintenance and for treatment of parasitic infections and cancers. On August 27, 2008, CDC was notified of a patient who developed hepatitis after a 1-week course of an herbal supplement containing artemisinin. The patient had abdominal pain, dark urine, and laboratory results consistent with hepatitis (e.g., serum alanine aminotransferase of 898 IU/L [normal: 10-55 IU/L]). Samples of the supplement were sent to CDC and the Georgia Institute of Technology for analysis to determine the amount of artemisinin and to identify any contaminants. Analysis indicated that the supplement contained 94%-97% of the 100 mg of artemisinin stated on the packaging and the supplement contained no other common pharmaceutical active ingredients. Given the patient's clinical course and laboratory evaluation, CDC investigators concluded that the hepatitis might have been associated with ingestion of the herbal supplement containing artemisinin. More data are needed to establish any causal connection between artemisinin and hepatitis. Health-care providers should be aware of the possibility of hepatic toxicity in patients taking herbal supplements containing artemisinin. PMID- 19680222 TI - Greatwall maintains mitosis through regulation of PP2A. AB - Greatwall (GW) is a new kinase that has an important function in the activation and the maintenance of cyclin B-Cdc2 activity. Although the mechanism by which it induces this effect is unknown, it has been suggested that GW could maintain cyclin B-Cdc2 activity by regulating its activation loop. Using Xenopus egg extracts, we show that GW depletion promotes mitotic exit, even in the presence of a high cyclin B-Cdc2 activity by inducing dephosphorylation of mitotic substrates. These results indicate that GW does not maintain the mitotic state by regulating the cyclin B-Cdc2 activation loop but by regulating a phosphatase. This phosphatase is PP2A; we show that (1) PP2A binds GW, (2) the inhibition or the specific depletion of this phosphatase from mitotic extracts rescues the phenotype induced by GW inactivation and (3) the PP2A-dependent dephosphorylation of cyclin B-Cdc2 substrates is increased in GW-depleted Xenopus egg extracts. These results suggest that mitotic entry and maintenance is not only mediated by the activation of cyclin B-Cdc2 but also by the regulation of PP2A by GW. PMID- 19680223 TI - A non-canonical function of topoisomerase II in disentangling dysfunctional telomeres. AB - The decatenation activity of topoisomerase II (Top2), which is widely conserved within the eukaryotic domain, is essential for chromosomal segregation in mitosis. It is less clear, however, whether Top2 performs the same function uniformly across the whole genome, and whether all its functions rely on decatenation. In the fission yeast, Schizosaccharomyces pombe, telomeres are bound by Taz1, which promotes smooth replication fork progression through the repetitive telomeric sequences. Hence, replication forks stall at taz1 Delta telomeres. This leads to telomeric entanglements at low temperatures (10-fold and virtually block the translocation of larger ones. This suggests that passive and facilitated NPC passage proceed through one and the same permeability barrier. PMID- 19680229 TI - Thymus-specific deletion of insulin induces autoimmune diabetes. AB - Insulin expression in the thymus has been implicated in regulating the negative selection of autoreactive T cells and in mediating the central immune tolerance towards pancreatic beta-cells. To further explore the function of this ectopic insulin expression, we knocked out the mouse Ins2 gene specifically in the Aire expressing medullary thymic epithelial cells (mTECs), without affecting its expression in the beta-cells. When further crossed to the Ins1 knockout background, both male and female pups (designated as ID-TEC mice for insulin deleted mTEC) developed diabetes spontaneously around 3 weeks after birth. beta cell-specific autoimmune destruction was observed, as well as islet-specific T cell infiltration. The presence of insulin-specific effector T cells was shown using ELISPOT assays and adoptive T cell transfer experiments. Results from thymus transplantation experiments proved further that depletion of Ins2 expression in mTECs was sufficient to break central tolerance and induce anti insulin autoimmunity. Our observations may explain the rare cases of type 1 diabetes onset in very young children carrying diabetes-resistant HLA class II alleles. ID-TEC mice could serve as a new model for studying this pathology. PMID- 19680230 TI - Individual and aggregate years-of-life-lost associated with overweight and obesity. AB - This study presents nationally representative estimates of individual and aggregate years-of-life-lost (YLLs) associated with overweight and three categories of obesity separately by age, race, smoking status, and gender strata. Using proportional hazards analysis and data from the National Health Interview Survey (NHIS) Linked Mortality Files, we estimated life expectancies for each BMI strata and quantified YLLs by comparing differences between each strata and the normal BMI reference group. Our results provide further evidence that overweight and mild obesity are not associated with a reduction in life expectancy. However, higher BMI categories are associated with lower expected survival. In aggregate, excess BMI is responsible for approximately 95 million YLLs. White females account for more than two-thirds of the aggregate YLLs. Unless something is done to reduce the rising prevalence of those with BMIs >35, or to mitigate the impact of obesity or its correlates on YLLs, expected life expectancy for US adults may decrease in the future. PMID- 19680231 TI - Associations of free fatty acids with insulin secretion and action among African American and European-American girls and women. AB - Ethnic differences in insulin secretion and action between African Americans (AAs) and European Americans (EAs) may influence mobilization of free fatty acids (FFAs). We tested the hypotheses that FFA concentrations would be associated with measures of insulin secretion and action before and during a glucose challenge test. Subjects were 48 prepubertal girls, 60 premenopausal women, and 46 postmenopausal women. Fasting insulin (insulin(0)), the acute insulin response to glucose (AIR(g)), the insulin sensitivity index (S(I)), basal and nadir FFA (FFA(0), FFA(nadir)), and nadir time (TIME(nadir)) were determined during an intravenous glucose tolerance test (IVGTT). Stepwise multiple linear regression (MLR) analysis was conducted to identify associations of FFA(0), FFA(nadir), and TIME(nadir) with ethnicity, age group, insulin measures, indexes of body composition from dual-energy X-ray absorptiometry, and measures of fat distribution from computed tomography scan. In this population, insulin(0) and AIR(g) were higher among AAs vs. EAs, whereas S(I) was lower, independent of age group. MLR analyses indicated that FFA(0) was best predicted by lean tissue mass (LTM), leg fat mass, ethnicity (lower in AAs), S(I), and insulin(0). FFA(nadir) was best predicted by FFA(0), age group, and intra-abdominal adipose tissue (IAAT). TIME(nadir) was best predicted by leg fat mass, AIR(g), and S(I). In conclusion, indexes of insulin secretion and action were associated with FFA dynamics in healthy girls and women. Lower FFA(0) among AAs was independent of insulin(0) and S(I). Whether lower FFA(0) is associated with substrate oxidation or risk for obesity remains to be determined. PMID- 19680232 TI - Characterization of herpes virus entry mediator as a factor linked to obesity. AB - Herpes virus entry mediator (HVEM) is a member of the tumor necrosis factor (TNF) receptor superfamily (TNFRSF14), which serves as a receptor for herpes viruses and cytokines such as lymphotoxin-alpha (LT-alpha) and LIGHT (lymphotoxin-like inducible protein that competes with glycoprotein D for herpes virus entry on T cells). We aimed to explore the associations of HVEM with human obesity. HVEM gene expression and protein levels were studied in total adipose tissue and in their fractions (isolated adipocytes and stromovascular cells (SVCs)) obtained from 81 subjects during elective surgical procedures. HVEM -241GA and -14AG gene polymorphisms were also studied and associated with obesity measures in 840 subjects. Visceral adipose tissue had significantly higher expression of HVEM than subcutaneous adipose tissue (P < 0.0001). Obese patients had significantly higher subcutaneous HVEM gene expression (P = 0.03) and protein levels (P = 0.01) than lean subjects. HVEM gene expression and protein levels were found in both isolated adipocytes and SVCs. These findings were confirmed in primary cultures from human preadipocytes, in which a significant increase in HVEM was observed during the differentiation process. HVEM -241GA and -14AG gene polymorphisms were associated with obesity, diastolic pressure, several inflammatory parameters (C reactive protein and interleukin 18 (IL-18)), and circulating LIGHT concentrations. A sample of men with the G241A gene polymorphism also showed an increased serum titer of IgG antiherpes virus 1. These results provide evidences of an existing relationship between HVEM and obesity, which suggest that this TNF superfamily receptor could be involved in the pathogenesis of obesity and inflammation-related activity. PMID- 19680233 TI - Replication of association between a common variant near melanocortin-4 receptor gene and obesity-related traits in Asian Sikhs. AB - Recent genome-wide association studies (GWAS) in Asian Indians reported strong associations of variants near melanocortin-4 receptor (MC4R) and MLX interacting protein-like (MLXIPL) genes with insulin resistance and several obesity-related quantitative traits (QTs). Here, we evaluated the association of two variants (rs12970134 and rs4450508) near MC4R and a nonsynonymous (Gln241His) variant (rs3812316) in MLXIPL gene with type 2 diabetes (T2D) and obesity-related QTs in our case-control cohort (n = 1,528; 745 T2D cases and 783 controls) from a Sikh population from North India. We have successfully replicated the association of MC4R (rs12970134) with BMI (P = 0.0005), total weight (WT) (P = 0.001), and waist circumference (WC) (P = 0.001). These associations remained significant after controlling for multiple testing by applying Bonferroni's correction. However, our data did not confirm the association of rs3812316 in the MLXIPL gene with triglyceride (TG) levels. These observations demonstrate that the genetic variation in MC4R locus can have a moderate contribution in the regional fat deposition and development of central obesity in Asian Indians. PMID- 19680234 TI - Effects of catechin enriched green tea on body composition. AB - Obesity is a major health problem in the developed and developing world. Many "functional" foods and ingredients are advocated for their effects on body composition but few have consistent scientific support for their efficacy. However, an increasing amount of mechanistic and clinical evidence is building for green tea (GT). This experiment was therefore undertaken to study the effects of a high-catechin GT on body composition in a moderately overweight Chinese population. In a randomized placebo-controlled trial, 182 moderately overweight Chinese subjects, consumed either two servings of a control drink (C; 30 mg catechins, 10 mg caffeine/day), one serving of the control drink and one serving of an extra high-catechin GT1 (458 mg catechins, 104 mg caffeine/day), two servings of a high-catechin GT2 (468 mg catechins, 126 mg caffeine/day) or two servings of the extra high-catechin GT3 (886 mg catechins, 198 mg caffeine/day) for 90 days. Data were collected at 0, 30, 60, and 90 days. We observed a decrease in estimated intra-abdominal fat (IAF) area of 5.6 cm(2) in the GT3 group. In addition, we found decreases of 1.9 cm in waist circumference and 1.2 kg body weight in the GT3 group vs. C (P < 0.05). We also observed reductions in total body fat (GT2, 0.7 kg, P < 0.05) and body fat % (GT1, 0.6%, P < 0.05). We conclude that consumption of two servings of an extra high-catechin GT leads to improvements in body composition and reduces abdominal fatness in moderately overweight Chinese subjects. PMID- 19680235 TI - Association of long-term change in waist circumference with insulin resistance. AB - Recent studies have shown that fat accumulation is associated with insulin resistance; however, the risks associated with long-term changes and fluctuations in central fatness are less clear. This study examined the longitudinal relationship between waist circumference (WC) and insulin resistance using three dimensions of WC: baseline WC, slope of linear changes in WC, and fluctuation of WC around the slope during 20 years of follow-up. Anthropometry, insulin resistance (homeostasis model assessment (HOMA(IR))), and lifestyle factors were obtained in a population-based, prospective observational study (Coronary Artery Risk Development in Young Adults (CARDIA)) during 1985-2006, excluding participants who had been diagnosed with diabetes at any examination. After adjusting for socio-demographic and lifestyle factors, the evolution of HOMA(IR) from CARDIA year 15 to 20 was 6.9% higher per standard deviation of year 0 WC (P trend <0.0001) and 6.3% higher per standard deviation increase in the change in WC over the long term (P trend <0.0001). However, WC fluctuations around the linear change were not associated with insulin resistance or its evolution. The level of HOMA(IR) increased substantially with steeper linear WC slope among initially thinner participants at baseline, whereas this association tended to be weaker in those with higher initial WC (P interaction <0.0001). We conclude that year 0 WC and long-term increment in WC are associated with worsening insulin resistance. However, the association of HOMA(IR) with slope of WC change may vary across the range of initial WC. PMID- 19680236 TI - Increased expression of DNA methyltransferase 3a in obese adipose tissue: studies with transgenic mice. AB - Epigenetic mechanisms are likely to be involved in the development of obesity. This study was designed to examine the role of a DNA methyltransferase (Dnmt3a), in obese adipose tissue. The gene expression of Dnmts was examined by quantitative real-time PCR analysis. Transgenic mice overexpressing Dnmt3a in the adipose tissue driven by the aP2 promoter were created (Dnmt3a mice). DNA methylation of downregulated genes was examined using bisulfite DNA methylation analysis. Dnmt3a mice were fed a methyl-supplemented or high-fat diet, and subjected to body weight measurement and gene expression analysis of the adipose tissue. Expression of Dnmt3a was markedly upregulated in the adipose tissue of obese mice. The complementary DNA (cDNA) microarray analysis of Dnmt3a mice revealed a slight decrease in the gene expression of secreted frizzled-related protein 1 (SFRP1) and marked increase in that of interferon responsive factor 9 (IRF9). In the SFRP1 promoter, DNA methylation was not markedly increased in Dnmt3a mice relative to wild-type mice. In experiments with a high-fat diet or methyl-supplemented diet, body weight did not differ significantly with the genotypes. Gene expression levels of inflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha) and monocyte chemoattractant protein-1 (MCP-1) were higher in Dnmt3a mice than in wild-type mice on a high-fat diet. This study suggests that increased expression of Dnmt3a in the adipose tissue may contribute to obesity-related inflammation. The data highlight the potential role of Dnmt3a in the adult tissue as well as in the developing embryo and cancer. PMID- 19680237 TI - Treatment of obese adolescents: the influence of periodization models and ACE genotype. AB - The aims of the present study were to compare the effects of two periodization models on metabolic syndrome risk factors in obese adolescents and verify whether the angiotensin-converting enzyme (ACE) genotype is important in establishing these effects. A total of 32 postpuberty obese adolescents were submitted to aerobic training (AT) and resistance training (RT) for 14 weeks. The subjects were divided into linear periodization (LP, n = 16) or daily undulating periodization (DUP, n = 16). Body composition, visceral and subcutaneous fat, glycemia, insulinemia, homeostasis model assessment of insulin resistance (HOMA IR), lipid profiles, blood pressure, maximal oxygen consumption (VO(2max)), resting metabolic rate (RMR), muscular endurance were analyzed at baseline and after intervention. Both groups demonstrated a significant reduction in body mass, BMI, body fat, visceral and subcutaneous fat, total and low-density lipoprotein cholesterol, blood pressure and an increase in fat-free mass, VO(2max), and muscular endurance. However, only DUP promoted a reduction in insulin concentrations and HOMA-IR. It is important to emphasize that there was no statics difference between LP and DUP groups; however, it appears that there may be bigger changes in the DUP than LP group in some of the metabolic syndrome risk factors in obese adolescents with regard to the effect size (ES). Both periodization models presented a large effect on muscular endurance. Despite the limitation of sample size, our results suggested that the ACE genotype may influence the functional and metabolic characteristics of obese adolescents and may be considered in the future strategies for massive obesity control. PMID- 19680238 TI - Effects of doubling the portion size of fruit and vegetable side dishes on children's intake at a meal. AB - Increasing the portion size of energy-dense entrees has been shown to increase children's energy intake during a meal. It remains to be investigated whether serving larger portions to children can be used to promote intake of more healthful foods, such as fruits and vegetables (F&V). The aim of the present study was to examine the effects of increasing the portion size of F&V side dishes on children's intake. Forty-three children (22 boys, 21 girls), aged 5-6 years, were served dinner once a week for 2 weeks. Each dinner consisted of pasta with tomato sauce, three F&V side dishes (broccoli, carrots, and applesauce), and milk. The portion size of the F&V was doubled between experimental conditions whereas the size of the pasta remained constant. Doubling the portion size of the side dishes resulted in a 43% increase in children's intake of the fruit side dish (P = 0.001), but did not affect children's intake of the two vegetable side dishes (P > 0.60). Further, when the portion size of F&V side dishes was doubled, children ate significantly less of the pasta (P = 0.04). The difference in meal energy intake between portion size conditions (19.5 +/- 16.3 kcal) was not significant (P = 0.24). Although more studies are needed to understand whether increases in portion size can influence vegetable intake, children did eat more in response to a large quantity of a preferred low energy-dense fruit side dish at meals. Thus variations in portion size can be used strategically to help children achieve the recommended intake of fruits. PMID- 19680239 TI - Structures of the tRNA export factor in the nuclear and cytosolic states. AB - Transfer RNAs are among the most ubiquitous molecules in cells, central to decoding information from messenger RNAs on translating ribosomes. In eukaryotic cells, tRNAs are actively transported from their site of synthesis in the nucleus to their site of function in the cytosol. This is mediated by a dedicated nucleo cytoplasmic transport factor of the karyopherin-beta family (Xpot, also known as Los1 in Saccharomyces cerevisiae). Here we report the 3.2 A resolution structure of Schizosaccharomyces pombe Xpot in complex with tRNA and RanGTP, and the 3.1 A structure of unbound Xpot, revealing both nuclear and cytosolic snapshots of this transport factor. Xpot undergoes a large conformational change on binding cargo, wrapping around the tRNA and, in particular, binding to the tRNA 5' and 3' ends. The binding mode explains how Xpot can recognize all mature tRNAs in the cell and yet distinguish them from those that have not been properly processed, thus coupling tRNA export to quality control. PMID- 19680240 TI - Designing and using synthetic RNA thermometers for temperature-controlled gene expression in bacteria. AB - Many techniques have been developed for studying inducible gene expression, but all of them are multicomponent systems consisting of cis-acting elements at the DNA or RNA level, trans-acting regulator proteins and/or small molecules as inducers. RNA thermometers are the only known single-component regulators of gene expression. They consist of a temperature-sensitive secondary structure in the 5' untranslated region of the mRNA, which contains the ribosome-binding site. The ribosome-binding site can be masked or unmasked by a simple temperature shift, thereby repressing or inducing translation. Recently, we and others have designed synthetic RNA thermometers that are considerably simpler than naturally occurring thermometers and can be exploited as convenient on/off switches of gene expression. In this protocol, we describe the construction and use of synthetic RNA thermometers. We provide guidelines for the in silico design of thermometer controlled mRNA leaders and for their experimental testing and optimization; the entire procedure can be completed in 2-3 weeks. PMID- 19680241 TI - m-TAG: a PCR-based genomic integration method to visualize the localization of specific endogenous mRNAs in vivo in yeast. AB - This protocol describes m-TAG, a novel method for the visualization of endogenously expressed mRNAs in live yeast (Saccharomyces cerevisiae). First, a gene of interest is tagged with multiple binding sites for the RNA-binding MS2 coat protein (MS2-CP), using a PCR-based genomic-tagging strategy and homologous recombination. Next, MS2-CP fused to GFP(x3) is expressed in cells; binding of this fusion protein to the tagged mRNA yields an RNA granule that can be visualized by fluorescence microscopy. While existing methods necessitate cell fixation (for in situ hybridization) or the detection of exogenously expressed mRNAs (from plasmids), or give transient signals (i.e., with fluorescent hybridization probes), m-TAG allows for the robust and stable visualization of endogenously expressed mRNAs in vivo and facilitates the study of mRNA dynamics under different growth conditions. The m-TAG procedure is simple, easy to perform and takes <3 weeks to yield cultured yeast strains for mRNA visualization. PMID- 19680242 TI - Drosophila melanogaster as a model host for studying Pseudomonas aeruginosa infection. AB - Conservation of host signaling pathways and tissue physiology between Drosophila melanogaster and mammals allows for the modeling of human host-pathogen interactions in Drosophila. Here we present the use of genetically tractable Drosophila models of bacterial pathogenesis to study infection with the human opportunistic pathogen Pseudomonas aeruginosa. We describe and compare two protocols commonly used to infect Drosophila with P. aeruginosa: needle-pricking and injector-pumping. Each model has relevance for examining host components and bacterial factors in host defense and virulence. Fly survival and bacterial proliferation within host flies can be assessed as a measure of host susceptibility and pathogen virulence potential. The profiles of host responses toward P. aeruginosa virulent and non-virulent strains can be determined, enabling the identification of interaction-specific genes that could potentially favor or limit the initiation and progression of infection. Both of the protocols presented herein may be adapted for the inoculation and study of other microbial pathogens. P. aeruginosa cell preparation requires 24 h, fly inoculation 1 h, and fly survival and bacterial proliferation 1-4 d. PMID- 19680243 TI - A macrodomain-containing histone rearranges chromatin upon sensing PARP1 activation. AB - Poly-ADP-ribosylation is a post-translational modification catalyzed by PARP enzymes with roles in transcription and chromatin biology. Here we show that distinct macrodomains, including those of histone macroH2A1.1, are recruited to sites of PARP1 activation induced by laser-generated DNA damage. Chemical PARP1 inhibitors, PARP1 knockdown and mutation of ADP-ribose-binding residues in macroH2A1.1 abrogate macrodomain recruitment. Notably, histone macroH2A1.1 senses PARP1 activation, transiently compacts chromatin, reduces the recruitment of DNA damage factor Ku70-Ku80 and alters gamma-H2AX patterns, whereas the splice variant macroH2A1.2, which is deficient in poly-ADP-ribose binding, does not mediate chromatin rearrangements upon PARP1 activation. The structure of the macroH2A1.1 macrodomain in complex with ADP-ribose establishes a poly-ADP-ribose cap-binding function and reveals conformational changes in the macrodomain upon ligand binding. We thus identify macrodomains as modules that directly sense PARP activation in vivo and establish macroH2A histones as dynamic regulators of chromatin plasticity. PMID- 19680244 TI - Efficient targeting of expressed and silent genes in human ESCs and iPSCs using zinc-finger nucleases. AB - Realizing the full potential of human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) requires efficient methods for genetic modification. However, techniques to generate cell type-specific lineage reporters, as well as reliable tools to disrupt, repair or overexpress genes by gene targeting, are inefficient at best and thus are not routinely used. Here we report the highly efficient targeting of three genes in human pluripotent cells using zinc-finger nuclease (ZFN)-mediated genome editing. First, using ZFNs specific for the OCT4 (POU5F1) locus, we generated OCT4-eGFP reporter cells to monitor the pluripotent state of hESCs. Second, we inserted a transgene into the AAVS1 locus to generate a robust drug-inducible overexpression system in hESCs. Finally, we targeted the PITX3 gene, demonstrating that ZFNs can be used to generate reporter cells by targeting non-expressed genes in hESCs and hiPSCs. PMID- 19680245 TI - From small reads do mighty genomes grow. AB - This month's Genome Watch discusses the use of next-generation sequencing technologies to assemble draft genomes for two pseudomonad species. PMID- 19680247 TI - Waves of resistance: Staphylococcus aureus in the antibiotic era. AB - Staphylococcus aureus is notorious for its ability to become resistant to antibiotics. Infections that are caused by antibiotic-resistant strains often occur in epidemic waves that are initiated by one or a few successful clones. Methicillin-resistant S. aureus (MRSA) features prominently in these epidemics. Historically associated with hospitals and other health care settings, MRSA has now emerged as a widespread cause of community infections. Community or community associated MRSA (CA-MRSA) can spread rapidly among healthy individuals. Outbreaks of CA-MRSA infections have been reported worldwide, and CA-MRSA strains are now epidemic in the United States. Here, we review the molecular epidemiology of the epidemic waves of penicillin- and methicillin-resistant strains of S. aureus that have occurred since 1940, with a focus on the clinical and molecular epidemiology of CA-MRSA. PMID- 19680252 TI - Franklin Harold Epstein: 5 May 1924-5 November 2008. PMID- 19680248 TI - Bacterial cell division: assembly, maintenance and disassembly of the Z ring. AB - Bacterial cell division is orchestrated by a tubulin homologue, FtsZ, which polymerizes to form a ring-like structure that is both a scaffold for the assembly of the bacterial cytokinetic machinery and, at least in part, a source of the energy for constriction. FtsZ assembly is tightly regulated, and a diverse repertoire of accessory proteins contributes to the formation of a functional division machine that is responsive to cell cycle status and environmental stress. In this Review, we describe the interaction of these proteins with FtsZ and discuss recent advances in our understanding of Z ring assembly. PMID- 19680250 TI - The precursors of memory: models and controversies. AB - The adaptive immune system has evolved a unique capacity to remember a pathogen through the generation of memory T cells, which rapidly protect the host in the event of reinfection. How memory T cells develop and the relationship between effector and memory T cells has been actively debated in the literature for many years and several models have been proposed to explain the divergent developmental fates of T cell progeny. Here, Nature Reviews Immunology asks four leading researchers in the field to provide their thoughts and opinions on the ontogeny of memory T cells and its implications for vaccine design. PMID- 19680254 TI - When is one kidney not enough? AB - Most renal failure in children results from congenital anomalies of the kidney and urinary tract (CAKUTs). Sanna-Cherchi et al. predict that by 30 years of age, nearly 50% of patients with a solitary kidney would be receiving dialysis. This outcome differs markedly from that of renal-transplant donors, who have no increased risk for renal failure. Because morbidity from CAKUTs may not develop until adulthood, these patients should be closely followed throughout life. PMID- 19680256 TI - Recalcitrant pain in a patient with ADPKD. PMID- 19680249 TI - The type III secretion system of Pseudomonas aeruginosa: infection by injection. AB - The Gram-negative bacterium Pseudomonas aeruginosa uses a complex type III secretion apparatus to inject effector proteins into host cells. The configuration of this secretion machinery, the activities of the proteins that are injected by it and the consequences of this process for infection are now being elucidated. This Review summarizes our current knowledge of P. aeruginosa type III secretion, including the secretion and translocation machinery, the regulation of this machinery, and the associated chaperones and effector proteins. The features of this interesting secretion system have important implications for the pathogenesis of P. aeruginosa infections and for other type III secretion systems. PMID- 19680255 TI - A novel classification for IgA nephropathy. AB - IgA nephropathy is a common glomerulonephritis. About 30% of patients develop end stage kidney disease within 20 years. A strategy for predicting prognosis from biopsy-based pathological evaluation has not been well established because of lack of reproducibility and inappropriate categorization. The new Oxford classification of IgA nephropathy provides a histopathological grading system with reproducibility for prediction of renal prognosis of IgA nephropathy independent of the clinical features. The classification must be validated in the different cohorts. PMID- 19680257 TI - Toward revealing the roles for intermedin in the community of vasoactive peptides. PMID- 19680258 TI - Intermedin is upregulated and has protective roles in a mouse ischemia/reperfusion model. AB - Intermedin (IMD), a new calcitonin/calcitonin gene-related peptide family peptide with vasodilatory and positive inotropic properties, has multiple functions in regulating cardiovascular homeostasis and is of particular interest in the pathophysiology of myocardial ischemia/reperfusion (MI/R). We created a mouse model of MI/R by ligating the cardiac left anterior descending artery to study the possible pathophysiological role of IMD and its receptor complexes in MI/R. Compared with the control, infarcted mice showed increased content, mRNA and protein expression of IMD in plasma and cardiac tissue. The mRNA expression of the receptor activity-modifying protein 3 (RAMP3) gene increased very early, and the calcitonin receptor-like receptor and RAMP2 mRNA levels increased later after reperfusion. However, the RAMP1 gene expression did not change. The tissue IMD content was positively correlated with the diastolic blood pressure and negatively correlated with pulse pressure. In addition, exogenous IMD treatment significantly ameliorated the MI/R injury by rescuing the pulse pressure, inhibiting neutrophil infiltration in the peri-infarction area, and decreasing the creatine kinase and lactate dehydrogenase activities in plasma. Our results indicated that IMD was upregulated in the ischemic myocardium and may induce important beneficial cytoprotection against cardiac ischemic injury. PMID- 19680259 TI - Prevention of local lipotoxicity: a new renoprotective mechanism of peroxisome proliferator-activated receptor-alpha activation in hypertension and obesity? PMID- 19680260 TI - Impact of azelnidipine treatment on left ventricular diastolic performance in patients with hypertension and mild diastolic dysfunction: multi-center study with echocardiography. AB - We investigated the impact of lowering blood pressure (BP) with azelnidipine, a newly developed calcium channel blocker, generation on the left ventricular (LV) diastolic function and LV filling pressure by assessing non-invasive indices derived from echo Doppler study. This study evaluated 232 hypertensive patients with diastolic dysfunction. This study had two groups: (1) in which azelnidipine was administered to patients as a first-line therapy, and (2) in which amlodipine was converted to azelnidipine. Early diastolic mitral annulus velocity (e', cm s( 1)), the ratio of peak E velocity to e' velocity (E/e' ratio) and level of brain natriuretic peptide (BNP) were measured before and, an average of, 8 months after azelnidipine treatment. In the first-line azelnidipine group, the systolic and diastolic BP reduced by 26 and 11 mm Hg, respectively. The e' increased, and E/e' ratio and BNP level decreased significantly. In the converted-from-amlodipine group, the systolic and diastolic BP decreased by 14 and 6 mmHg, respectively. The e' velocity increased, but the E/e' ratio and BNP level did not change. In both groups, azelnidipine lowered BP and improved LV diastolic function (an increase in the e' velocity). Possible reduction in LV filling pressure (a decrease in the E/e' ratio and BNP level) is observed only in the first-line azelnidipine group. PMID- 19680262 TI - Regulation of TNFRSF and innate immune signalling complexes by TRAFs and cIAPs. AB - There have been a number of recent discoveries relating to the functions of inhibitors of apoptosis (IAPs) and TNF receptor-associated factors (TRAFs) in regulating signalling from TNF receptor superfamily (TNFRSF) members and some tantalizing glimpses into a wider area of influence, that of innate immune signalling. Discoveries relating to the function of these ubiquitin E3 ligases in regulating signalling from the eponymous member of the family, TNF-R1, are dealt with superbly in a separate review by Wertz and Dixit and so we will confine our discussion to the subset of the TNFRSF that does not contain a death domain (DD). In line with the available data we will divide the review into two parts, the first is restricted to the role of TRAFs 2 and 3 and cIAPs in regulating TNFRSF signalling, whereas the second will be more speculative, asking what role IAPs and TRAFs have in innate immune signalling. PMID- 19680261 TI - Loss of function of DJ-1 triggered by Parkinson's disease-associated mutation is due to proteolytic resistance to caspase-6. AB - DJ-1 was recently identified as a gene product responsible for a subset of familial Parkinson's disease (PD). The mechanisms by which mutations in DJ-1 alter its function and account for PD-related pathology remained largely unknown. We show that DJ-1 is processed by caspase-6 and that the caspase-6-derived C terminal fragment of DJ-1 fully accounts for associated p53-dependent cell death. In line with the above data, we show that a recently described early-onset PD associated mutation (D149A) renders DJ-1 resistant to caspase-6 proteolysis and abolishes its protective phenotype. Unlike the D149A mutation, the L166P mutation that prevents DJ-1 dimerization does not impair its proteolysis by caspase-6 although it also abolishes DJ-1 antiapoptotic function. Therefore, we show here that DJ-1 loss of function could be due to impaired caspase-6 proteolysis and we document the fact that various DJ-1 mutations could lead to PD pathology through distinct molecular mechanisms. PMID- 19680263 TI - Monomeric CRP contributes to complement control in fluid phase and on cellular surfaces and increases phagocytosis by recruiting factor H. AB - Complement forms the first defense line of innate immunity and has an important role in the non-inflammatory clearance of apoptotic and necrotic cells. Factor H is one essential complement inhibitor that binds to the acute phase reactant C reactive protein (CRP). By using recombinant proteins, calcium-independent binding of Factor H to monomeric CRP (mCRP), but not to pentameric CRP (pCRP), was shown. In addition to the two known CRP-binding sites, a novel third site was localized within the C-terminus. This region is frequently mutated in the hemolytic uremic syndrome and the mutant proteins show reduced mCRP binding. In this study, we show that mCRP directs Factor H to the surface of apoptotic and necrotic endothelial cells and identify phosphocholine as one binding moiety for this complex. Factor H-mCRP complexes enhance C3b inactivation both in the fluid phase and on the surface of damaged cells and inhibit the production of pro inflammatory cytokines. By recruiting the soluble complement inhibitor Factor H to the surface of damaged cells, mCRP blocks the progression of the complement cascade beyond the step of the C3 convertase, prevents the formation of inflammatory activation products, and thus contributes to the safe removal of opsonized damaged cells and particles. PMID- 19680264 TI - Dissecting the role of ubiquitylation in the DNA damage response checkpoint in G2. AB - Maintenance of genomic integrity is one of the fundamental biological properties shared by all living organisms. To counterbalance deleterious and potentially mutagenic effects of omnipresent DNA damaging assaults, organisms have developed a network of genome surveillance and maintenance pathways known as the DNA damage response. In eukaryotes, the orchestration of cell-cycle checkpoints, DNA damage repair, and apoptosis in response to DNA damage relies on posttranslational modifications of key regulatory proteins. Although the role of phosphorylation in these pathways is relatively well established, the significance of ubiquitylation has only recently emerged. In this review, we survey current research on the ubiquitin-proteasome system, focusing on the DNA damage response in the G2 phase of the cell cycle and two prominent classes of ubiquitin ligases, the SCF- and APC/C complexes. These ubiquitin ligases are reviewed with regard to their function in activating, maintaining, and terminating the checkpoint and in light of increasing evidence that suggests a dynamic balance of substrate ubiquitylation and deubiquitylation. We further discuss the impact of defective G2 checkpoint signaling on genomic stability and cancer risk, highlighting strategies for targeted antitumor drug discovery. PMID- 19680265 TI - Hax1 lacks BH modules and is peripherally associated to heavy membranes: implications for Omi/HtrA2 and PARL activity in the regulation of mitochondrial stress and apoptosis. AB - Hax1 has an important role in immunodeficiency syndromes and apoptosis. A recent report (Chao et al., Nature, 2008) proposed that the Bcl-2-family-related protein, Hax1, suppresses apoptosis in lymphocytes and neurons through a mechanism that involves its association to the inner mitochondrial membrane rhomboid protease PARL, to proteolytically activate the serine protease Omi/HtrA2 and eliminate active Bax. This model implies that the control of cell-type sensitivity to pro-apoptotic stimuli is governed by the PARL/Hax1 complex in the mitochondria intermembrane space and, more generally, that Bcl-2-family-related proteins can control mitochondrial outer-membrane permeabilization from inside the mitochondrion. Further, it defines a novel, anti-apoptotic Opa1-independent pathway for PARL. In this study, we present evidence that, in vivo, the activity of Hax1 cannot be mechanistically coupled to PARL because the two proteins are confined in distinct cellular compartments and their interaction in vitro is an artifact. We also show by sequence analysis and secondary structure prediction that Hax1 is extremely unlikely to be a Bcl-2-family-related protein because it lacks Bcl-2 homology modules. These results indicate a different function and mechanism of Hax1 in apoptosis and re-opens the question of whether mammalian PARL, in addition to apoptosis, regulates mitochondrial stress response through Omi/HtrA2 processing. PMID- 19680267 TI - CD95 engagement mediates actin-independent and -dependent apoptotic signals. AB - CD95 is a death receptor whose stimulation by either the physiologic ligand CD95L or the agonistic antibodies leads to the formation of a multi-molecular complex termed DISC (death-inducing signaling complex) and the subsequent induction of a caspase-driven apoptotic signal. According to the magnitude of the DISC formation, two types of cells have been identified. Although type I cells generate an important DISC, the complex is barely found in type II cells. Analyzing the early stages preceding the DISC formation, we found that unlike CD95L, the commonly used agonistic antibody APO1-3 internalized the death receptor. Using inhibitors of actin polymerization, we showed that the remodeling of the actin cytoskeleton did not alter the capping of the CD95 receptor or its partitioning into the lipid rafts. In addition, whereas the disruption of F-actin prevented the internalization of CD95, the DISC formation and the apoptotic signal induced by the agonistic antibody APO1-3 in type I cells, it did not affect the signal triggered by the soluble and membrane-bound CD95L, regardless of the type of cells. In conclusion, the addition of APO1-3 on type I cells triggers an actin-dependent apoptotic signal, which is absent or marginal in cells (both types I and II) treated with CD95L. PMID- 19680266 TI - TRPC channel-mediated neuroprotection by PDGF involves Pyk2/ERK/CREB pathway. AB - Platelet-derived growth factor-BB (PDGF) has been reported to provide tropic support for neurons in the central nervous system. The protective role of PDGF on dopaminergic neurons, especially in the context of HIV-associated dementia (HAD), however, remains largely unknown. Here, we show that exogenous PDGF was neuroprotective against toxicity induced by HIV-1 Tat in primary midbrain neurons. Furthermore, we report the involvement of transient receptor potential canonical (TRPC) channels in PDGF-mediated neuroprotection. TRPC channels are Ca(2+)-permeable, nonselective cation channels with a variety of physiological functions. Blocking TRPC channels with either a blocker or short-interfering RNAs (specific for TRPC 5 and 6) in primary neurons resulted in suppression of both PDGF-mediated neuroprotection as well as elevations in intracellular Ca(2+). PDGF mediated neuroprotection involved parallel but distinct ERK/CREB and PI3K/Akt pathways. TRPC channel blocking also resulted in suppression of PDGF-induced Pyk2/ERK/CREB activation, but not Akt activation. Relevance of these findings in vivo was further corroborated by intrastriatal injections of PDGF and HIV-1 Tat in mice. Administration of PDGF was able to rescue the dopaminergic neurons in the substantia nigra from Tat-induced neurotoxicity. This effect was attenuated by pre-treatment of mice with the TRP blocker, thus underscoring the novel role of TRPC channels in the neuroprotection mediated by PDGF. PMID- 19680268 TI - Ancestry informative markers and admixture proportions in northeastern Mexico. AB - To investigate the ancestral admixture in the Mestizo population in northeastern Mexico, we genotyped 74 ancestral informative markers (AIMs) and 15 Y-single nucleotide polymorphisms (Y-SNPs) in 100 individuals. The Native American contribution is 56% (range: 27.4-81.2%), the European contribution is 38% (range: 16.7-70.5%) and the West African contribution is 6%. The results show a higher European contribution than was reported in other similar studies in the country, albeit with a predominant Native American ancestry. No remarkable differences in the ancestry proportions were observed using subgroups of 74, 54, 34 and 24 AIMs. The paternal lineage calculated by genotyping of 15 Y-SNPs, shows a major component of European and Eurasian ancestry markers ( approximately 78%), compared with Amerindian ( approximately 12%) and African markers (10%). This information will set a reference for future determinations of admixture proportions in the Mestizo population from Mexico and for population-based association studies of complex diseases. PMID- 19680271 TI - Dramatic visual recovery after prompt radiotherapy and chemotherapy for leukaemic infiltration of the optic nerve in a child. PMID- 19680269 TI - Validation of the association between AGTRL1 polymorphism and coronary artery disease in the Japanese and Korean populations. AB - Coronary artery disease (CAD) and stroke are the major health problems in many countries because of their increasing prevalence and high mortality. It is well known that CAD and stroke are based on atherosclerosis and shared environmental and genetic risk factors. Recently, an association of a functional sequence variation -154G>A in the angiotensin receptor-like 1 (AGTRL1) with a susceptibility to stroke was reported. In this study, we investigated a total of 1479 CAD cases and 2062 controls from the Japanese and Korean populations to validate the association of AGTRL1 with CAD. However, we obtained no evidence of the association in both the Japanese (odds ratio (OR)=0.95, 95% confidence interval (CI); 0.82-1.10, P=0.47, allele count model) and Korean (OR=0.90, 95% CI; 0.77-1.05, P=0.18, allele count model) populations. In addition, there was no trend of association between the risk allele and severity of coronary atherosclerosis. These data suggested that AGTRL1 did not contribute much to the atherosclerosis of the coronary artery. PMID- 19680270 TI - Identification of novel candidate loci for anorexia nervosa at 1q41 and 11q22 in Japanese by a genome-wide association analysis with microsatellite markers. AB - The Japanese Genetic Research Group for Eating Disorders (JGRED) is a multisite collaborative study group that was organized for the systematic recruitment of patients with an eating disorder for the purpose of genetic study in Japan. We conducted a genome-wide case-control association study using 23 465 highly polymorphic microsatellite (MS) markers to identify genomic loci related to anorexia nervosa (AN). Pooled DNA typing in two screening stages, followed by individual typing of 320 AN cases and 341 controls, allowed us to identify 10 MS markers to be associated with AN. To narrow down genomic regions responsible for the association of these MS markers, we further conducted a single-nucleotide polymorphism (SNP) association analysis for 7 of the 10 loci in 331 AN cases and 872 controls, which include the 320 AN cases and the 341 controls genotyped in the MS screening, respectively. Two loci, namely 1q41 and 11q22, remained significantly associated with AN in the SNP-based fine mapping, indicating the success in narrowing down susceptibility regions for AN. Neither of these loci showed a positive evidence of association with bulimia nervosa. The most significant association was observed at SNP rs2048332 (allelic P-value=0.00023) located at 3'-downstream of the SPATA17 gene on the 1q41 locus. The association analysis for MS-SNP haplotypes detected a statistically significant association (permutation P-value=0.00003) of the A-4-G-T haplotype that comprised four SNP/MS markers (rs6590474-D11S0268i-rs737582-rs7947224) on the 11q22 locus with AN. This linkage disequilibrium block spanning a 20.2-kb interval contains exon 9 of the CNTN5 gene encoding contactin 5. PMID- 19680272 TI - Learning effect in visual field testing of healthy subjects using Humphrey Matrix frequency doubling technology perimetry. AB - PURPOSE: To evaluate the learning effect of frequency doubling technology (FDT) perimetry using the Humphrey Matrix perimeter in healthy subjects with no perimetric experience. METHODS: One eye of 28 healthy adults who had no history of visual field testing underwent three Matrix tests using 24-2 programme with full-threshold strategy. The results of the first session were compared with those of the second, and the third sessions. Learning effect was defined as an improvement at results for duration, perimetric indices, and the number of points with a P<5 and <1% in the total and pattern deviation maps. Anderson's criteria were applied to define abnormal examinations. RESULTS: The mean in the mean deviation global index was -7.40+/-1.49 dB in the first session, -4.64+/-0.97 dB in the second session, and -3.29+/-0.93 dB in the third session. Pattern standard deviations were decreased as the tests were repeated (P<0.01). Test duration, fixation losses, false negative, and the number of points with a P<5 and <1% in the total and pattern deviation maps rate were also changed significantly (P<0.05). Among the criteria suggested by Anderson, the Glaucoma Hemifield Test performed better in all session tests. The false-positive rates ranged from 50% (95% confidence intervals [CI] 30.6-69.4%) to 64% (95% CI 44.1-81.4%) for the first session, 29% (95% CI 13.2-48.7%) to 54% (95% CI 33.9-72.5%) for the second session, and from 18% (95% CI 6.1-36.9%) to 32% (95% CI 15.9-52.4%) for the third session. CONCLUSIONS: The results of this study show that the learning effect for Humphrey Matrix FDT perimetry must be considered in normal individuals with no perimetric experience. PMID- 19680273 TI - ARMS2/HTRA1 and CFH polymorphisms are not associated with choroidal neovascularization in highly myopic eyes of the elderly Japanese population. AB - PURPOSE: The purpose of this study was to investigate whether the genetic risk factors of age-related macular degeneration (AMD) are associated with the development of choroidal neovascularization (CNV) in highly myopic eyes of elderly Japanese. METHODS: Highly myopic elderly Japanese patients with and without CNV were genotyped for three AMD-associated single nucleotide polymorphisms (SNPs), namely rs10490924 (A69S) of ARMS2, rs11200638 of HTRA1, and rs1061170 (Y402H) of complement factor H (CFH), with the TaqMan SNP assay. One hundred and eighty-three unrelated highly myopic (axial lengths>26.00 mm or refractive errors>-6.0 diopters) Japanese patients with CNV who were >or=50 years of age (mean age+/-standard deviation of 62.7+/-6.3 years) and 170 highly myopic patients without CNV who were >or=50 years old (62.3+/-7.1 years) were studied. The differences in the genotypic distributions for the three SNPs between the two groups were tested with the Trend chi2 test, and logistic regression analyses were performed for age and gender adjustment. RESULTS: No significant difference was detected in the distribution of the three SNPs, rs10490924 (P>0.1), rs11200638 (P>0.1), and rs1061170 (P>0.5), between the two groups even after adjustments for age and gender differences. CONCLUSION: The genetic risk factors of AMD related to these SNPs do not contribute significantly to the development of CNV in a highly myopic elderly Japanese population. PMID- 19680274 TI - Functional and morphological changes of macula after subthreshold micropulse diode laser photocoagulation for diabetic macular oedema. AB - PURPOSE: To examine the early changes of retinal sensitivity by fundus-related microperimetry after subthreshold micropulse diode laser photocoagulation (SMDLP) for diabetic macular oedema (DMO). METHODS: Twenty-eight eyes of 28 patients with diffuse DMO were treated with SMDLP. The mean retinal sensitivity within the central 10 degrees measured with a fundus-related microperimeter, MP1, best corrected visual acuity (BCVA) in logarithm of minimum angle of resolution units, and optical coherence tomography-determined foveal thickness (FT) were examined before and 3 months after SMDLP. The pretreatment values of the retinal sensitivity, FT, BCVA, and funduscopic findings were compared with the corresponding values at 3 months after SMDLP. RESULTS: At 3 months, the BCVA was significantly improved (P=0.03), and the FT was significantly reduced (P=0.0043). The mean retinal sensitivity within the central 10 degree, however, did not change significantly (P=0.70). The correlation between the changes in the retinal sensitivities and the decrease in the FT was not significant. The correlation between the changes in the retinal sensitivities and the BCVA was also not significant. CONCLUSIONS: Significant improvements in retinal sensitivities within the central 10 degrees were not observed even though the decrease in FT and the improvement of BCVA were significant. On account of this difference of changes in retinal sensitivity and BCVA, the combination of retinal sensitivity by MP1 and BCVA may be beneficial in assessing the visual function from various angles after SMDLP for eyes with DMO. PMID- 19680276 TI - Ocular findings in sleep apnoea patients using continuous positive airway pressure. AB - PURPOSE: To describe ocular findings in patients with established obstructive sleep apnoea hypopnoea syndrome (OSAHS) using continuous positive airway pressure (CPAP). METHODS: hundred and fifteen referrals investigated for OSAHS were included. Patients with OSAHS were compared with those with normal sleep study controls. Subgroup analysis for CPAP users and non-users was also carried out. RESULTS: OSAHS patients (n=89) compared with the controls (n=26) had higher ocular irritation symptoms (P<0.001), abnormal tear break-up time (P<0.05) with increased upper (P<0.001) and lower (P<0.001) lid laxity. Floppy eyelid syndrome (FES) was noted in 31.5% (28/89) OSAHS patients vs 3.8% (1/26) controls (P=0.005). Open angle glaucoma prevalence in OSAHS patients (3/89, 3.4%) was similar to the controls (1/26, 3.8%) (P=0.92). Sixty-seven (75.3%) OSAHS patients were using CPAP (average duration: 19.6+/-15.3 months). All CPAP users maintained a supine sleep posture to prevent mask edge leaks. A fifth of CPAP users (14/67) had experienced earlier episodes of conjunctivitis secondary to leaks. CPAP users had similar upper and lower lid laxity (P=0.746 and 0.633) to non-CPAP users, but a better tear film (P=0.029) and less ocular irritation (P=0.134). CONCLUSION: OSAHS patients showed increased ocular irritation, abnormal tear film, lid laxity, and FES. The prevalence of glaucoma in our series was similar to normal population data of 2%, P=0.429, and may relate to use of CPAP in majority of the patients. More stable tear film in CPAP users was probably secondary to the supine sleep postures necessarily adopted with CPAP use. PMID- 19680275 TI - Comparison of 25- and 23-gauge sutureless microincision vitrectomy surgery in the treatment of various vitreoretinal diseases. AB - PURPOSE: To compare the effectiveness and safety of 25- and 23-gauge sutureless microincision vitrectomy surgery (MIVS) in the management of various vitreoretinal diseases. METHODS: Eighty-five consecutive patients undergoing sutureless vitrectomy during January to April 2008 were randomized to either 25- or 23-gauge MIVS. Data collected prospectively included best-corrected visual acuity (BCVA), operation time, postoperative visual recovery, postoperative anterior segment change, and complications. RESULTS: The most common indications for MIVS were macular hole, macular pucker, vitreous haemorrhage, and diabetic traction retinal detachment. Mean operation times of the 25-gauge (n=38) and 23 gauge groups (n=47) were 33.68 and 34.47 min, respectively (P=0.942). Mean BCVA improved significantly in both the 25- and 23-gauge groups when measured 3-month postoperatively. There was no between-group difference in either the degree or the rate of postoperative visual recovery. Seven patients in the 23-gauge group, compared with three in the 25-gauge group, required suturing of sclerotomy at the end of the surgery. No patients in either group developed postoperative wound leakage or endophthalmitis. CONCLUSION: Our prospective study suggests that, within the limited indications, both 25- and 23-gauge MIVS are equally effective, with similar safety profiles. Gauge selection thus may be made according to a surgeon's preference and the availability of appropriate instruments. PMID- 19680277 TI - Effectiveness of iron repletion in the diet for the optic nerve development of anaemic rats. AB - PURPOSE: To compare the process of myelination in the developing optic nerve (ON) of anaemic rats with the subsequent recovery after being fed an iron-recovery diet. METHODS: In this study, the morphometrical parameters in the ON were assessed by electron microscopy in Wistar rats that were on an iron-deficient diet for 32 days or for 21 days followed by 10 days on an iron-recovery diet. Qualitative and quantitative analyses were performed using representative electron ultramicrographs. Data were analysed by one-way analysis of variance (ANOVA). When differences were detected, comparisons were made using Tukey's post hoc test (P<0.05 was considered to be significant). RESULTS: Qualitative analysis of the ONs in anaemic and recovered animals showed a higher rate of deformed axons and increased lamellar separation in the myelin sheath when compared with the respective control group. The ON of the anaemic group showed a reduced mean density of myelinated fibres when compared with the control group. The fibre area ratio, axon area ratio, and myelin area ratio of large axons/small axons in the ONs of the control group showed the highest values for the myelin areas, axon areas, and total fibre areas. The control group showed a significantly higher myelin sheath thickness when compared with the anaemic and recovered groups. CONCLUSIONS: Our data indicate that iron is necessary for maintenance of the ON cell structure, and that morphological damage from iron deficiency is not easily reverted by iron repletion. PMID- 19680278 TI - The Cataract National Dataset electronic multicentre audit of 55,567 operations: when should IOLMaster biometric measurements be rechecked? AB - PURPOSE: Calculation of intraocular lens (IOL) power for implantation during cataract surgery depends on ocular biometric measurements. The aim of this study was to characterise the normal range of intra- and interindividual variation in axial length (AL) and corneal power (K) when IOLMaster measurements were possible and to derive recommendations as to which outlying measurements merit verification before acceptance. METHODS: The Medisoft electronic patient database contains prospectively collected data conforming to the United Kingdom (UK) Cataract National Dataset on 55,567 cataract operations. From this AL and K information on the 32,556 eyes (14,016 paired) of patients older than 25 years, without corneal pathology, history of intraocular surgery and who had all biometric measurements taken with the Zeiss IOLMaster (Carl Zeiss Meditec) were extracted. R 2.8.1 (R Foundation for Statistical Computing) was used for statistical analysis. RESULTS: Mean age was 76.4 years and 62.0% were female. Mean (95% confidence interval) values for AL, mean K and corneal astigmatism were 23.40 (21.27-26.59) mm, 43.90 (40.94-47.01) D and 1.04 (<2.50) D. Nearly all astigmatism was either with or against the rule. Differences between paired eyes were not statistically significant. 95% individuals had asymmetry of AL and mean K<0.70 mm and 0.92 D, respectively. CONCLUSIONS: On the basis of approximation of the 95% CI above, it is suggested that AL, mean K and keratometric astigmatism measurements outside the ranges 21.30-26.60 mm, 41.00-47.00 D and >2.50 D, respectively, and intraindividual asymmetry of AL >0.70 mm or mean K>0.90 D should be verified before acceptance. PMID- 19680279 TI - Intravitreal bevacizumab (Avastin) for age-related macular degeneration: a critical analysis of literature. AB - PURPOSE: The current medical environment demands that quality health care is delivered at an affordable cost through the use of objective, unbiased clinical data. This study was undertaken to review the current literature on bevacizumab for age-related macular degeneration and its value in determining best clinical practice. METHODS: Randomised controlled trials (RCTs) and observational studies that met the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) criteria were identified from the current literature for further analysis. Data concerning treatment dosing regimens, response to treatment, complications, and factors influencing outcome and safety were extracted and compiled into a database. RESULTS: As of January 2009, there were 5 RCTs that compared the outcomes of bevacizumab to other treatment options and 50 studies that met the STROBE criteria with similar visual and anatomical outcomes between RCTs and observational studies. Although the doses and dosing frequencies varied between the studies, the mean gain in vision at 3 months was +7.76+/-5.4 ETDRS letters (range +2 to +14.4); an effect that was maintained at 6 months in studies with longer follow-up. Predominantly classic lesions were the most responsive of all lesion subtypes. The complication profiles/rates were similar to those reported with other anti-vascular endothelial agents. CONCLUSIONS: There is sufficient scientific and statistical evidence to advocate the effective use of OCT-guided administration of intravitreal bevacizumab for neovascular AMD. This is reflected in our study outcome measures that are comparable to findings published from recent well-conducted RCTs on intravitreal ranibizumab at the same time point. PMID- 19680280 TI - The Cataract National Dataset electronic multi-centre audit of 55,567 operations: variation in posterior capsule rupture rates between surgeons. AB - AIMS: To demonstrate variations in posterior capsule rupture (PCR) rate between surgeons of the same and different grades as a by-product of routine clinical care. METHOD: NHS departments using electronic medical record (EMR) systems to collect the Cataract National Dataset (CND) were invited to submit data. Data were remotely extracted, anonymised, assessed for conformity and completeness, and analysed for rates of PCR for individual surgeons within each of the three grades. RESULTS: Data were extracted on 55,567 cataract operations performed at 12 NHS trusts by 406 surgeons between November 2001 and July 2006. Data on the grade of 404 of the 406 surgeons who contributed to the study were available for 55,515 cases (99.9%) and were used for this analysis. Variation in PCR rate between surgeons was highest for the most junior grade of surgeon and between those surgeons contributing relatively few cases to the data set. Variation in PCR was lowest among experienced surgeons contributing large numbers of cases to the data set. CONCLUSIONS: Considerable variation in PCR rate exists both between and within surgical grades. Routine electronic collection of the CND allows detailed analysis of variations in PCR rates between individual surgeons. To define acceptable limits for this benchmark complication of cataract surgery, further work is needed to adjust surgeons' outcomes for the case mix complexity. PMID- 19680281 TI - Patterns of progression of localized retinal nerve fibre layer defect on red-free fundus photographs in normal-tension glaucoma. AB - PURPOSES: To investigate patterns of progression of localized retinal nerve fibre layer (RNFL) defect on red-free fundus photographs and to quantify extents of progression in normal-tension glaucoma (NTG) patients. METHODS: Sixty-five eyes of consecutive 65 NTG patients who had shown progression of localized RNFL defect on serial red-free fundus photographs were selected for this study. Patterns of progression of localized RNFL defect on red-free fundus photographs were categorized and extents of progression were quantified. Serial assessments of disc stereophotographs and visual fields were also performed to detect progression. RESULTS: The most common pattern of progression was widening of the defect towards the macula (n=37; 56.9%) followed by deepening of the defect (n=25; 38.5%), appearance of a new defect (n=6; 9.2%), and widening of the defect away from the macula (n=5; 7.7%). Eight eyes simultaneously showed two patterns of progression. Mean angular widening of the defect towards the macula and away from the macula was 6.4+/-4.1 degrees (range: 1.1-17.1 degrees , n=37) and 3.4+/ 2.1 degrees (range: 1.1-5.2 degrees , n=5), respectively. No progression was observed on the disc stereophotographs (n=65) or in the visual fields (n=55) in 64 eyes (98.5%) and 46 eyes (83.6%), respectively. CONCLUSIONS: There were four patterns of progression of localized RNFL defect. In most cases, RNFL loss proceeded temporally. PMID- 19680282 TI - New beta-class milbemycin compound from Streptomyces avermitilis NEAU1069: fermentation, isolation and structure elucidation. PMID- 19680283 TI - TLN-05220, TLN-05223, new Echinosporamicin-type antibiotics, and proposed revision of the structure of bravomicins(*). AB - The deposited strain of the hazimicin producer, Micromonospora echinospora ssp. challisensis NRRL 12255 has considerable biosynthetic capabilities as revealed by genome scanning. Among these is a locus containing both type I and type II PKS genes. The presumed products of this locus, TLN-05220 (1) and TLN-05223 (2), bear a core backbone composed of six fused rings starting with a 2-pyridone moiety. The structures were confirmed by conventional spectral analyses including MS, and 1D and 2D NMR experiments. Comparison of both the 1H and 13C NMR data of the newly isolated compound with those of echinosporamicin and bravomicin A led us to propose a revision of the structure of the latter to include a 2-pyridone instead of the pyran originally postulated. Both compounds (1 and 2) possessed strong antibacterial activity against a series of gram-positive pathogens including several strains of methicillin-resistant Staphylococcus aureus and vancomycin resistant Enterococci (VRE), and cytotoxic activities against several human tumor cell lines. The TLN compounds are the first of this group with reported anticancer activity. PMID- 19680284 TI - LL-Z1272alpha epoxide, a precursor of ascochlorin produced by a mutant of Ascochyta viciae. AB - A novel metabolite, LL-Z1272alpha epoxide, structurally related to ascochlorin, was isolated from the cultured mycelium of Ascochyta viciae J-29, a mutant derived from A. viciae Libert. The structure was elucidated on the basis of spectroscopic data. The epoxide is proposed to be enzymatically formed from LL Z1272alpha and is a precursor of ascochlorin, an antiviral and antitumor antibiotic. The conversion of the epoxide to ascochlorin by cyclization of its farnesyl chain to a cyclohexanone ring is similar to that of squalene 2, 3-oxide to sterols. Unlike ascochlorin, the new metabolite had no growth inhibitory activity against Candida albicans in the paper-disc agar diffusion assay. PMID- 19680286 TI - Melanoma-initiating cells: a compass needed. AB - Most tumours contain a heterogeneous population of cancer cells, which harbour a range of genetic mutations and have probably undergone deregulated differentiation programmes that allow them to adapt to tumour microenvironments. Another explanation for tumour heterogeneity might be that the cells within a tumour are derived from tumour-initiating cells through diverse differentiation programmes. Tumour-initiating cells are thought to constitute one or more distinct subpopulations within a tumour and to drive tumour initiation, development and metastasis, as well as to be responsible for their recurrence after therapy. Recent studies have raised crucial questions about the nature, frequency and importance of melanoma-initiating cells. Here, we discuss our current understanding of melanoma-initiating cells and outline several approaches that the scientific community might consider to resolve the controversies surrounding these cells. PMID- 19680287 TI - Bub1 and Bub3 promote the conversion from monopolar to bipolar chromosome attachment independently of shugoshin. AB - The eukaryotic spindle assembly checkpoint (SAC) delays anaphase in the presence of chromosome attachment errors. Bub3 has been reported to be required for SAC activity in all eukaryotes examined so far. We find that Bub3, unlike its binding partner Bub1, is not essential for the SAC in fission yeast. As Bub3 is needed for the efficient kinetochore localization of Bub1, and of Mad1, Mad2 and Mad3, this implies that most SAC proteins do not need to be enriched at the kinetochores for the SAC to function. We find that Bub3 is also dispensable for shugoshin localization to the centromeres, which is the second known function of Bub1. Instead, Bub3, together with Bub1, has a specific function in promoting the conversion from chromosome mono-orientation to bi-orientation. PMID- 19680288 TI - Pervasive transcription constitutes a new level of eukaryotic genome regulation. AB - During the past few years, it has become increasingly evident that the expression of eukaryotic genomes is far more complex than had been previously noted. The idea that the transcriptome is derived exclusively from protein-coding genes and some specific non-coding RNAs--such as snRNAs, snoRNAs, tRNAs or rRNAs--has been swept away by numerous studies indicating that RNA polymerase II can be found at almost any genomic location. Pervasive transcription is widespread and, far from being a futile process, has a crucial role in controlling gene expression and genomic plasticity. Here, we review recent findings that point to cryptic transcription as a fundamental component of the regulation of eukaryotic genomes. PMID- 19680289 TI - The structure of bacterial RNA polymerase in complex with the essential transcription elongation factor NusA. AB - There are three stages of transcribing DNA into RNA. These stages are initiation, elongation and termination, and they are well-understood biochemically. However, despite the plethora of structural information made available on RNA polymerase in the last decade, little is available for RNA polymerase in complex with transcription elongation factors. To understand the mechanisms of transcriptional regulation, we describe the first structure, to our knowledge, for a bacterial RNA polymerase in complex with an essential transcription elongation factor. The resulting structure formed between the RNA polymerase and NusA from Bacillus subtilis provides important insights into the transition from an initiation complex to an elongation complex, and how NusA is able to modulate transcription elongation and termination. PMID- 19680290 TI - Induction of RNA-directed DNA methylation upon decondensation of constitutive heterochromatin. AB - Centromeric constitutive heterochromatin is marked by DNA methylation and dimethylated histone H3 Lys 9 (H3K9me2) in Arabidopsis. RNA-directed DNA methylation (RdDM) is a process that uses 24-nucleotide (nt) small interfering RNAs (siRNAs) to induce de novo methylation to its homologous DNA sequences. Despite the presence of centromeric 24-nt siRNAs, mutations in genes required for RdDM do not appreciably influence the methylation of centromeric repeats. The mechanism by which constitutive heterochromatin is protected from RdDM remains puzzling. Here, we report that the vegetative cell nuclei (VN) of the male gametophyte (pollen) invariably undergo extensive decondensation of centromeric heterochromatin and lose centromere identity. VN show greatly reduced H3K9me2, phenocopying nuclei carrying a mutation in the chromatin remodeller DECREASE IN DNA METHYLATION 1 (DDM1). However, unlike the situation in ddm1 nuclei, the decondensed heterochromatin retains dense CG methylation and transcriptional silencing, and, unexpectedly, is subjected to RdDM-dependent hypermethylation in non-CG contexts. These findings reveal two assembly orders of silent heterochromatin and implicate the condensed form in blocking the RdDM machinery. PMID- 19680291 TI - The role of genetic testing in the prediction of response to EGFR inhibitors in NSCLC. AB - Lung cancer is the leading cause of cancer deaths worldwide. Non-small-cell lung cancer (NSCLC) is the largest subgroup of the disease and accounts for approximately 80% of lung cancers. Most patients with NSCLC are diagnosed with locally advanced or metastatic disease. The current treatment options for such patients are associated with substantial limitations in efficacy and safety. Targeted therapies have provided some improvement in clinical outcomes. Recent efforts have focused on identifying specific markers that are predictive of response to treatment. A pretreatment detection of such markers could facilitate a more personalized and specific approach to therapy, whereby the most appropriate and efficacious treatment is selected for a specific subset of patients. Such a tailored approach would maximize both the therapeutic index and cost-effectiveness of treatments. In addition, several novel agents are under development, which may have the potential for overcoming acquired resistance to existing treatments. This supplement will review the current and emerging treatments for patients with advanced NSCLC and the molecular predictors of response that may facilitate the future clinical application of personalized medicine. PMID- 19680292 TI - Overview of molecular testing in non-small-cell lung cancer: mutational analysis, gene copy number, protein expression and other biomarkers of EGFR for the prediction of response to tyrosine kinase inhibitors. AB - Most patients with non-small-cell lung cancer (NSCLC) present with advanced disease. Current treatment paradigms are shifting from cytotoxic chemotherapies alone to single-agent and combination biological and targeted therapies. As patient responses to these therapies vary, predictive biomarkers will be an important facet of a patient's diagnostic workup in personalized medicine, as there is accumulating evidence that they may enable the prognostication and prediction of therapeutic response. Potential biomarkers for the selection of patients with NSCLC most likely to benefit from epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs), such as gefitinib and erlotinib, include mutations, gene copy number increase and single-nucleotide polymorphisms of the EGFR gene, EGFR protein expression and oncogenic mutation on the KRAS gene. Many techniques are available to assay for these biomarkers. In this review, we present the current weight of evidence for using these methods as biomarkers for anti-EGFR therapy in patients with NSCLC. PMID- 19680294 TI - Predictive value of EGFR and HER2 overexpression in advanced non-small-cell lung cancer. AB - Epidermal growth factor receptor (EGFR) and HER2 are cell surface receptor tyrosine kinases (TKs) that transduce growth signals through dimerization with HER family receptors. The heterodimerization of EGFR with HER2 induces a more potent activation of EGFR TK than does EGFR homodimerization. When tumor cells overexpress both EGFR and HER2, they exhibit aggressive tumor cell growth, owing to the increased potential for EGFR/HER2 heterodimerization and signaling. Gefitinib and erlotinib are EGFR TK inhibitors (EGFR TKIs) and have antitumor activity in 8-18% of patients with advanced non-small-cell lung cancer (NSCLC). Certain patient subsets are particularly responsive to EGFR TKIs. Analyses of biomarkers from patients in clinical studies of EGFR TKIs show correlations between objective tumor response and EGFR overexpression, as detected by immunohistochemistry and increased gene copy number measured by fluorescence in situ hybridization analysis. Furthermore, NSCLC tumors that overexpress both EGFR and HER2 are more sensitive to EGFR TKIs than are tumors that overexpress EGFR but are HER2 negative. Therefore, the measurement of EGFR and HER2 protein expression and the gene copy number in NSCLC tumors may have a prognostic value in NSCLC and a predictive value for identifying patients likely to benefit from an EGFR TKI. These considerations suggest that the simultaneous inhibition of EGFR and HER2 may warrant further study in patients with NSCLC. PMID- 19680295 TI - Individualized therapy in non-small-cell lung cancer: future versus current clinical practice. AB - Despite advances in the management of non-small-cell lung cancer (NSCLC), including the introduction of targeted therapies such as epidermal growth factor receptor tyrosine kinase inhibitors, improvements in survival are marginal and the overall prognosis for patients remains poor. Tailoring therapy to the individual patient is a promising approach for selecting the most appropriate therapeutic regimens to maximize efficacy and minimize toxicity. The identification of predictive biomarkers that can guide treatment decisions is an important step for individualized therapy and in ultimately improving patient outcomes. Genomic and proteomic studies provide a means for the molecular profiling of tumor tissue from patients with NSCLC, and allow tailoring of therapy whereby the most appropriate treatment is administered to each individual patient. Although there are still significant challenges to implementing genomic and proteomic testing in clinical practice, the rapid development of newer technologies provides hope for overcoming these barriers. PMID- 19680293 TI - Activating and resistance mutations of EGFR in non-small-cell lung cancer: role in clinical response to EGFR tyrosine kinase inhibitors. AB - The epidermal growth factor receptor tyrosine kinase inhibitors (EGFR TKIs), gefitinib and erlotinib, are reversible competitive inhibitors of the tyrosine kinase domain of EGFR that bind to its adenosine-5' triphosphate-binding site. Somatic activating mutations of the EGFR gene, increased gene copy number and certain clinical and pathological features have been associated with dramatic tumor responses and favorable clinical outcomes with these agents in patients with non-small-cell lung cancer (NSCLC). The specific types of activating mutations that confer sensitivity to EGFR TKIs are present in the tyrosine kinase (TK) domain of the EGFR gene. Exon 19 deletion mutations and the single-point substitution mutation L858R in exon 21 are the most frequent in NSCLC and are termed 'classical' mutations. The NSCLC tumors insensitive to EGFR TKIs include those driven by the KRAS and MET oncogenes. Most patients who initially respond to gefitinib and erlotinib eventually become resistant and experience progressive disease. The point mutation T790M accounts for about one half of these cases of acquired resistance. Various second-generation EGFR TKIs are currently being evaluated and may have the potential to overcome T790M-mediated resistance by virtue of their irreversible inhibition of the receptor TK domain. PMID- 19680296 TI - Shortcomings of current therapies for non-small-cell lung cancer: unmet medical needs. AB - Non-small-cell lung cancer (NSCLC) is a major global health problem and is the leading cause of cancer death worldwide. Current treatment involves nonspecific, nonselective cytotoxic chemotherapy, which results in only a modest increase in survival and causes significant toxicity to the patient. Targeted agents are initially effective in certain small subpopulations of patients, but eventually nearly all patients become resistant to further treatment. The limitations in efficacy and safety associated with available treatments for NSCLC underscore the need for novel agents with improved efficacy and safety profiles. This review discusses the limitations of currently recommended therapies for patients with advanced NSCLC and discusses new agents in clinical development for this disease. PMID- 19680297 TI - Driveway accidents in New Zealand. PMID- 19680298 TI - Immunisation in hospital--an opportunity repeatedly missed. PMID- 19680299 TI - Influenza A(H1N1)09: another public health risk to New Zealand. PMID- 19680300 TI - Paediatric driveway run-over injuries: time to redesign? AB - AIMS: To investigate the demographic, accident, and environmental characteristics associated with driveway run-over injuries in order to identify potentially modifiable factors and prevention strategies METHODS: Retrospective review of all children less than 15 years old who were hospitalised or killed due to a driveway run-over injury in the Auckland region of New Zealand over the 50-month period, November 2001 to December 2005. Data was collected on the demographics, accident and environmental characteristics, and parental awareness. Data was obtained from clinical records and telephone interviews with parents. RESULTS: A total of 93 cases were identified, including 9 fatalities. The median age was 2 years with 73% under 5 years old. Children of Pacific Island and Maori ethnicity comprised 43% and 25% of cases respectively. Injuries occurred predominately on the child's home driveway (80% of cases). In 64% the driveway was the usual play area for the child. Only 13% of driveways were fenced. 51% were long driveways extending through the section, and 51% were shared with other properties. 51% of properties were rented and of these 57% were government houses. CONCLUSION: The absence of physical separation between driveways and children's play and living areas may predispose to driveway injuries. Further research is needed to investigate the ideal way to implement such separation in current properties and future property developments. PMID- 19680301 TI - Opportunistic immunisation of paediatric inpatients at Rotorua Hospital: audit and discussion. AB - AIM: To audit current practice around opportunistic immunisation in a New Zealand hospital and make recommendations for improvement. METHODS: We reviewed inpatient notes for 369 patients aged 3-23 months admitted over a 6-month period in 2007. Data was obtained regarding children's immunisation status, documentation of this and action taken in response to the under-immunised child. Literature review and discussion with local clinicians were used to identify recommendations for future practice. RESULTS: 84% of patients had their immunisation status documented on admission; only 60% of these were up-to-date with immunisations. Official immunisation records were rarely available. In 79% of patients who were behind with immunisation, no follow-up action was recorded, and only 4% of under immunised children received catch-up immunisation on the unit. CONCLUSIONS: Current practice around opportunistic immunisation is poor. A number of measures could be expected to improve this; these include establishing routine systems for obtaining immunisation records, a visual reminder system for immunisation and training more staff in immunisation. PMID- 19680302 TI - Preventing winter falls: a randomised controlled trial of a novel intervention. AB - AIM: To investigate the hypothesis that wearing socks over shoes improves traction on icy footpaths. METHODS: Randomised controlled trial involving 30 pedestrians (median age 21 years, range 18-70) travelling in a downhill direction on icy public footpaths at two sites in Dunedin, New Zealand. INTERVENTION: different coloured socks applied over normal footwear or usual practice (unadulterated footwear). PRIMARY OUTCOME: difference in mean self-reported slipperiness on a 5-point scale. SECONDARY OUTCOMES: falls, observer-rated slipperiness, observer-rated confidence, time to descend study slope. RESULTS: Two-thirds of participants (65%) had previously fallen on ice. Wearing socks over normal footwear was associated with a statistically significant improvement in traction; the difference in mean self-reported slipperiness scores between the control (n=15) and intervention (n=14) groups was 1.3 (95%CI: 0.4-2.3). Agreement between self-rated and observer-rated slipperiness was high (r=0.70). A higher proportion of the intervention group (71% vs 53%) appeared confident. One member of the control group fell. There was no evidence of risk compensation in the intervention group (difference in mean descent times 1.9 seconds, 95%CI: -6.1 10.0). The only adverse events were short periods of indignity for some members of the intervention group. CONCLUSION: Wearing socks over shoes appears to be an effective and inexpensive method to reduce the likelihood of slipping on icy footpaths. PMID- 19680303 TI - Infant and perinatal outcomes of triplet pregnancy in Auckland: better than expected? AB - AIM: There were two aims to the study: (1) to provide local outcome data that would be useful in counselling prospective parents of triplets; and (2) to address the deficit in accurate contemporary data on neurodevelopmental outcome and neonatal morbidity for those infants weighing less than 1500 g at birth. METHODS: We reviewed the outcome of triplet pregnancies born at National Women's Hospital / Auckland City Hospital (Auckland, New Zealand) for 1995-2005 inclusive. For this study triplet pregnancy was defined as a pregnancy beyond 20 weeks leading to registration of at least one birth. RESULTS: For the study period, 55 triplet pregnancies were identified. Forty-five percent of the pregnancies were reported as spontaneously conceived and 60% had no major complications other than premature delivery. One pregnancy spontaneously aborted; three fetuses from one pregnancy were stillborn, and four infants died in delivery suite. The median gestational age at birth was 32 (23-37) weeks and birth weight 1620 (530-2780) g. The median (range) Apgar score, for liveborns, was 8 (2-10) and 10 (4-10) for 1 and 5 minutes respectively. There were five neonatal deaths. Fifty-three infants, <1500g at birth, underwent formal developmental assessment. Three had cerebral palsy (2 hemiplegia and 1 spastic diplegia); one had marked motor delay and one hearing impairment requiring aids. The median Bayley II MDI was 95 (71-105) and PDI 94 (65-110). Outcomes were categorised in surviving triplets <1500 g as normal in 66%, mild abnormality in 17%, moderate abnormality in 15% and severely abnormal in only 2%. CONCLUSION: Although triplets represent a significant burden on the regional NICUs the outcome, including those <1500 g at birth, compares favourably with that reported. PMID- 19680304 TI - Pacific Islands Families: Child and Parental Physical Activity and Body Size- design and methodology. AB - AIM: To objectively assess physical activity (PA) and body size in 6-year-old children and their mothers participating in the Pacific Islands Families (PIF) cohort study, and to identify factors potentially related to PA and body size in Pacific children. METHODS: The PIF cohort was drawn from live births at Middlemore Hospital (South Auckland, New Zealand) in 2000. Information has been collected at birth, 6 weeks, 12 and 24 months, and 4 and 6 years postpartum. At 6 years, the Child and Parental Physical Activity and Body Size (PIF:PAC) study was simultaneously conducted and measures of child and mother PA (8-day accelerometry), body size (waist circumference, body mass index), and PA supports and barriers (questionnaire) taken. RESULTS: 254 mothers and their children took part in the PIF:PAC study. Usable accelerometer data were gathered for 173 mothers and 199 children over an average of 3-4 days. High levels of overweight and obesity were found in boys, girls, and mothers (62%, 58%, and 97% overweight or obese, respectively). CONCLUSION: Strategies for obesity reduction in Pacific populations are urgently required. Combined, the PIF and PIF:PAC studies will provide vital information for understanding and targeting the obesity epidemic in Pacific children. PMID- 19680305 TI - Sudden unexpected infant death and bedsharing: referrals to the Wellington Coroner 1997-2006. AB - AIMS: To describe the factors associated with sudden unexpected infant deaths, for which there was no clear medical diagnosis, referred to the Wellington-based coronial paediatric pathology service over the decade from 1997 to 2006. METHODS: The postmortem report, Police 47 file, Coroner's findings and deceased infant's medical records were used to create a profile for each sudden and unexpected infant death. RESULTS: There were 64 deaths in the period: 54 of these occurred during sleep and did not have a clear medical diagnosis. Maori and Pacific infants and infants from low decile areas were over-represented in the group. The majority (88.7%) of infants were < 6 months of age at death. Overall, 50% of infants had been placed to sleep in a non-recommended sleep position and 38% usually slept in a non-recommended location. Bedsharing was associated with 53.7% of deaths. There was a significant association between bedsharing and being found dead on a Sunday morning (p=0.04). CONCLUSION: Sudden unexpected death in infancy is associated with unsafe sleep environments and sleep positions. Every effort should be made to ensure that information about safe infant sleep practices reaches the caregivers of those particularly at risk. PMID- 19680306 TI - Acute infective conjunctivitis: evidence review and management advice for New Zealand practitioners. AB - In this review we present and discuss recent data and provide recommendations for New Zealand practitioners regarding the diagnosis and management of acute infective conjunctivitis. In particular, we discuss clinical predictors of bacterial versus viral conjunctivitis, a potential role for routine conjunctival culture, the benefits of topical antibiotic therapy for bacterial infections, delayed treatment algorithms, choice of topical antibiotic and the restriction of selected patients from work, school or early childhood care. PMID- 19680307 TI - A case of beta lactam-induced visual hallucination. PMID- 19680308 TI - Medical image. All that wheezes is not asthma. PMID- 19680309 TI - Use of placebos by New Zealand doctors. PMID- 19680310 TI - Response to letter from the NZ Council of Homeopaths. PMID- 19680311 TI - New software for modelling impacts of regional nuclear war: relevance to New Zealand. PMID- 19680312 TI - Determination of the speed of light by absolute wavelength measurement of the R(14) line of the CO2 9.4-Mum band and the known frequency of this line. PMID- 19680314 TI - Proposal for phase recovery from a single intensity distribution. PMID- 19680313 TI - Multicolor holographic imaging with a white-light source. PMID- 19680315 TI - Quasi fast Hankel transform. PMID- 19680317 TI - Spectroscopy of 3P0 states of alkaline earths. PMID- 19680316 TI - Amplified reflection, phase conjugation, and oscillation in degenerate four-wave mixing. PMID- 19680318 TI - Isotope-selective dissociation of the OsO4 molecule by two pulses of infrared radiation at different frequencies. PMID- 19680319 TI - Optoacoustic measurements of energy absorption in CO2 TEA-laser-excited SF6 at 293 and 145K. PMID- 19680320 TI - Transmission measurements of multiple photon absorption in SF6. PMID- 19680321 TI - Water-vapor-continuum absorption measurements (3.5 - 4.0 Mu m) using HDO-depleted water. PMID- 19680322 TI - Single-spindle aspheric figuring with flexible pads. PMID- 19680324 TI - Continuous tuning of 12 GHz in two bands of CO2 laser lines. PMID- 19680323 TI - Electro-optic multiplexer for large-numerical-aperture, low-loss fibers. PMID- 19680325 TI - GaAs laser array source package. PMID- 19680326 TI - Sidelobe suppression in corrugated-waveguide filters. PMID- 19680327 TI - High-resolution transient-double-resonance spectroscopy in SF6. AB - Four distinct collisionless excitation processes are observed when SF(6) molecules are illuminated simultaneously by "weak" continuously tunable cw probe radiation and by "strong" pulsed-pump radiation. These processes are identified by studying the line shape, frequency, and temporal decay of the change in transmitted probe intensity induced by the pump pulse. The significance of this work for the spectroscopy of high vibrational levels and for collisionless dissociation is discussed briefly. PMID- 19680328 TI - Interferometric two-dimensional imaging of rotating objects. AB - It is shown, theoretically and experimentally, that a rotating object can be two dimensionally imaged by illuminating the object with a sinusoidal interference pattern and then using the temporal modulation of the scattered light as the signal for building up a synthetic aperture. The image is formed in the Fourier transform plane of the synthetic aperture. PMID- 19680329 TI - White-light interferometry and phase-locked pulse analysis. AB - Two approaches to the analysis of spectral-temporal phenomena in optics have appeared recently. One utilizes channeling of white-light spectra and the other uses temporally resolved phase-locked pulses. We show that the two approaches are very closely related and that devices utilizing them have the same ultimate limits but quite different immediately practical limits. PMID- 19680330 TI - Generation of tunable continuous-wave ultraviolet radiation from 257 to 320 nm. AB - The generation of tunable cw radiation by sum-frequency mixing the output of a Rhodamine 6G laser and selected output lines of an argon-ion laser in an angle tuned KDP crystal has been investigated. This technique, coupled with second harmonic generation of the Rhodamine 6G laser, provides a simple source of cw ultraviolet radiation, continuously tunable from 257 to 320 nm, with a maximum output power in excess of 250 microW and a bandwidth of less than 0.03 nm. PMID- 19680331 TI - External-cavity-controlled 32-MHz narrow-band cw GaA1As-diode lasers. AB - By coupling a cw GaA1As-diode laser to an external resonator with Fabry-Perot etalons as dispersive elements, emission was reduced to a single-axial mode of 32 MHz width. The wavelength could be coarsely tuned over a spectral range of over 10 nm. Fine tuning over about 500 MHz was achieved by varying the external cavity length by less than lambda/3. At single-axial-mode operation, the commonly observed high- and low-frequency self-pulsing of the light output was found to disappear almost completely. PMID- 19680332 TI - Method of forming novel curved-line gratings and their use as reflectors and resonators in integrated optics. AB - Gratings have been used as Bragg reflectors in integrated optics for a variety of the applications. We report here a method of forming novel curved-line gratings and analyze their use for imaging in optical circuits and as the resonators in diode lasers. PMID- 19680333 TI - Design and fabrication of a blazed replica grating coupler for optical waveguides. AB - Replica grating couplers are useful for efficiently and inexpensively coupling a laser beam into a thin-film optical waveguide. Couplers have been designed, fabricated, and attached to optical waveguides. Input-coupling efficiencies as high as 75% have been measured on couplers having good uniformity over the entrance aperture. PMID- 19680334 TI - Spectrophone measurements of the absorption of visible light by aerosols in the atmosphere. AB - Simultaneous in situ measurements of the absorption and scattering by aerosols were made in ambient room air and in air with large amounts of added aerosols, such as cigarette smoke. For room aerosols, the extinction contribution due to absorption was typically one half that from scattering. The absorption measurements were made with an acoustically resonant spectrophone powered with a 0.5-W argon laser. PMID- 19680335 TI - Photon trapping and energy transfer in multiple-dye plastic matrices: an efficient solar-energy concentrator. AB - Experiments are described illustrating enhanced photon trapping and efficient energy transfer in mixed-dye planar solar concentrators containing, for example, Rhodamine 6G and Coumarin 6. These concentrators intercept more of the solar spectrum to give an enhanced photon-flux gain that exceeds the single-dye concentrator. It is also shown that the energy absorbed by the donor dye is transferred efficiently into the emitting acceptor by two competing processes. PMID- 19680336 TI - X-ray shadowgraphing in laser-produced plasma experiments. AB - We discuss a design for an x-ray framing camera. Shadowgraphing experiments using a laser-generated x-ray source demonstrate that 5-microm spatial resolution can be obtained for this camera with less than 7 J of laser energy to produce the x ray source. PMID- 19680337 TI - Precision measurements of NH(3) spectral lines near 11 microm using the infrared heterodyne technique. AB - Absolute line-center frequencies for eleven lines of ammonia in near coincidence with CO(2)-laser transitions have been determined to accuracies of +/-3 MHz by infrared heterodyne detection. These results were obtained by heterodyning a blackbody with a stable, grating-tuned CO(2) gas laser. A discussion of the apparatus and method of calculation, including error analysis, is presented. With these accurately determined line-center positions, the ammonia molecule will be a useful secondary-frequency standard for diode-laser spectroscopy in the 11-microm wavelength region. PMID- 19680338 TI - High-sensitivity infrared heterodyne radiometer using a tunable-diode-laser local oscillator. AB - A blackbody heterodyne radiometer using a widely tunable PbSnSe-diode laser as the local oscillator (LO) achieved signal-to-noise performance that was an order of magnitude better than previously reported and only a factor of 2.5 below that obtained with a CO(2)-laser LO. The diode laser system performance was within a factor of 6 of an ideal radiometer. High-resolution blackbody heterodyne absorption spectra of ethylene at 10.6 microm were obtained with a 0.4-sec post detection integration time by tuning the diode LO in a closed-cycle cryogenic cooler. PMID- 19680339 TI - Laser-pulse requirements for coherent and mode-selective excitation in the quasicontinuum of polyatomic molecules. AB - We show that the theory of nuclear magnetic relaxation applies with equal validity to intramolecular relaxation in the quasicontinuum of polyatomic molecules. This theory permits an analysis of the laser-pulse requirements for vibrational-mode selectivity and coherent multiphoton excitation. The laser Rabi precession frequency, microepsilon/h, should exceed 1/T(1) or 1/T(2), the intramolecular relaxation rates in the quasicontinuum. In practice this requires either an ultra-fast optical-pulse risetime or a slower pulse that is tuned far to the red side of the transition. PMID- 19680340 TI - Radiative spectral transfer in ruby. AB - The first direct observation of radiative spectral transfer within an inhomogeneously broadened line (the R(1) line of ruby) is reported. As opposed to nonradiative transfer, the temperature dependence of the radiative process is consistent with exponential, in agreement with recent predictions. A temperature independent, resonant process is also directly observed. The implication of these results for the concept of microscopic strain broadening is discussed. PMID- 19680341 TI - Saturation behavior of p-type germanium at CO(2) laser wavelengths. AB - Observed bleaching of single-crystal p-type germanium in the 10-microm region obeys an inhomogeneous broadening model for input intensities up to 100 times the saturation intensity I(s). Bleaching measurements show that I(s) varies from about 3.2 MW cm(-2) at 10.59 microm to about 6.8 MW cm(-2) at 9.27 microm. No significant variation of I(s) with crystal orientation is seen. Applications to CO(2) laser system isolation are discussed. PMID- 19680342 TI - A cw tunable near-infrared (1.085-1.175-microm) Raman oscillator. AB - This is the first report of a cw tunable Raman laser in the 1.1-microm region of the spectrum. A prism-tuned fiber Raman resonator with a 650-m-long, low-loss small-core single-mode silica fiber was pumped by a 5-W cw Nd:YAG laser at 1.064 microm. With two separate resonator mirrors for the first and the second Stokes oscillation, we have obtained first Stokes oscillation tunable from 1.085 to 1.13 microm and second Stokes oscillation tunable from 1.15 to 1.175 microm. PMID- 19680343 TI - Statistical properties of the sum of partially developed speckle patterns. AB - The probability density function and average contrast of the sum of n uncorrelated, partially developed speckle patterns have been theoretically investigated. A new form of the probability density function is given for the sum of n uncorrelated, partially developed speckle patterns under the assumption that the individual speckle fields to be added follow the circular statistics. PMID- 19680344 TI - Digital lock-in technique for measurement of polarization of radiation. AB - A highly versatile polarimeter has been constructed for determining all the Stokes parameters for a beam of radiation. Polarization measurements are performed using digital lock-in techniques by replacing the linear retardation plate with an electro-optic crystal, which can be externally modulated. The device lends itself readily to laboratory applications and provides the possibility of measuring small polarization quite accurately. PMID- 19680345 TI - Propagation of the mutual coherence function for an infinite plane wave through a turbid medium. AB - By means of a series of Fourier and inverse Fourier transformations, and by recourse to simple physical arguments concerning statistical stationarity and isotropy and the linearity of the scattering process, we are able to show that the propagation of the mutual coherence function for a plane wave through a turbid medium is governed by a pair of linear, first-order, one-dimensional, simultaneous differential equations. A sample solution to the equation is presented, and it is shown from a limiting form of this solution how the turbidity parameters in the differential equation can be obtained from a single scattering analysis. PMID- 19680346 TI - Full duplex transmission link over single-strand optical fiber. AB - Full duplex transmission of television signals over a single optical fiber is demonstrated. The transmission link uses directional couplers that possess high directivity and low insertion loss. PMID- 19680347 TI - Representation of codirectional coupled waves. AB - The evolution of the amplitudes of two general, codirectional coupled waves is represented as a rotation of a generalized Poincare sphere in analogy to polarized light. The method permits simple, pictorial representation of coupling effects in directional couplers and "monomode" optical fibers. PMID- 19680348 TI - High-sensitivity laser wavelength-modulation spectroscopy. AB - The method of doing dual-beam single-detector wavelength-modulation spectroscopy originally proposed by Bonfiglioli and Trench is demonstrated experimentally, using a tunable laser. This system monitors both the reference and the sample beams continuously in phase quadrature and measures the derivative and the direct spectra simultaneously. When this system was used in conjunction with an electro optically tuned cw dye laser, a sensitivity of (1/T) (dT/dlambda) = 3 x 10(-6) A( 1) was experimentally achieved for modulation depths from 1 to 50 A and can be extended down to 0.1 A. The derivative spectrum corresponding to the extremely weak fifth overtone of the C-H stretching vibration in the electronic ground state of benzene has been observed in a path length as short as 1 cm. PMID- 19680350 TI - Long-path high-resolution field measurements of absolute transmission in the 3.5- to 4.0-microm atmospheric window. AB - An example of high-resolution (0.08 cm(-1)), absolute transmission spectra of a 5 km atmospheric path is presented. Precise transmission and wavenumber calibration of the spectra were obtained from infrared laser extinction measurements. A comparison of these spectra to line-by-line calculations shows excellent agreement in the spectral region between 2480 and 2800 cm(-1) (3.57 to 4.03 microm). PMID- 19680349 TI - Optoacoustic pulses in a flame. AB - Pulsed pressure waves are produced by energy transfer from electronically excited Na or Li atoms seeded in an acetylene-air flame; the excitation is provided by a resonantly tuned, pulsed laser. Temperature increases of approximately 1 degree within the flame produce signals easily detected by a microphone. A description of the phenomenon is given, and preliminary results are presented for two possible applications: localized speed-of-sound measurements and quenching rate determinations. PMID- 19680351 TI - Unidirectional star coupler for single-fiber distribution systems. AB - The design and construction of a unidirectional star coupler for single-fiber distribution systems are described. A rectangular mixing section is required to obtain uniform output distribution. Four-port stars have been constructed with losses only (1/2) to 1 dB greater than packing fraction. For any given input, outputs are uniform to within about +/-1 dB. PMID- 19680352 TI - Determination of the surface structure autocorrelation function for rough deposits of magnesium. AB - Autocorrelation functions for rough surfaces of evaporated opaque magnesium deposits are determined from electron microphotographs of the shadowed-surface carbon replicas. These microphotographs are analyzed, using a data-processing system microdensitometer. It is shown that autocorrelation functions do not have a Gaussian form, as is usually assumed. PMID- 19680353 TI - Imaging with obscured pupils. AB - It is shown that diffraction-limited imaging systems with symmetric pupils and a noncentral obscuration yield a higher concentration of energy near the center of an image of a point object than those with central obscuration. PMID- 19680354 TI - Image blur for rainbow holograms. AB - An analysis is presented for selecting the important rainbow-hologram formation setup parameters for minimization of the image blur. PMID- 19680355 TI - Proposal for the determination of the complex degree of spatial coherence. AB - With the help of a folded-wavefront type of interferometer, a sinusoidally phase modulated wavefront can be superposed in parallel on another nonmodulated inverse wavefront. Spatial coherence is related to the fundamental and second-harmonic components of the modulation frequency included in the resultant intensity. PMID- 19680356 TI - Measuring fluid velocities with speckle patterns. AB - A new full-field technique for mapping lines of constant velocity in a fluid flow is demonstrated. The technique utilizes light scattered from scattering sites within a selected plane of interest of the flow field. The laser-speckle pattern thus formed is used with the established methods of speckle photography and interferometry to determine the velocity field. A double pulsed ruby laser is used to measure the velocity profile in a circular pipe. Turbulent flow fields will place a limitation on the technique; this limit is primarily a function of available equipment. PMID- 19680357 TI - Light scattering in converging beams. AB - Light scattering by spherical particles in a converging beam is obtained from the earlier solution for a particle in the beam of a dipole light source by reversing the direction of energy flow so that the dipole source becomes a sink. The results are not drastically different from Lorenz-Mie scattering as long as the sink is located at a distance greater than several particle radii from the focus. PMID- 19680361 TI - Absolute Raman frequency measurement of the Q(2) line in D(2) using cw CARS. AB - Coherent anti-Stokes Raman spectroscopy (CARS) is combined with high-resolution interferometry to measure the absolute Raman shift of the Q(2) vibrational line in D(2). The preliminary value found is 2987.237(1) +/- 0.001 cm(-1). Such precision is essential for the testing of ab initio energy-level calculations for the hydrogen isotopes. PMID- 19680362 TI - Stimulated Raman spectroscopy using low-power cw lasers. AB - It is demonstrated that stimulated Raman spectroscopy (SRS) can be performed using cw dye lasers at power levels over six orders of magnitude smaller than those generally associated with pulsed stimulated Raman studies. The preliminary results suggest that cw SRS is a potentially powerful alternative to conventional spontaneous Raman scattering, with resolution limited solely by laser linewidth and sensitivity independent of resolution requirements. PMID- 19680364 TI - Two-photon coherent propagation and third-harmonic generation. AB - Calculations of third-harmonic generation in conditions of coherent two-photon propagation in lithium vapor are presented that predict shapes, frequencies, and intensities for the fundamental pulse yielding an optimal third-harmonic energy conversion efficiency of 8%. PMID- 19680363 TI - Sensitivity enhancement of a spin-flip Raman laser absorption spectrometer through use of an intracavity absorption cell. AB - Gas-phase molecular spectroscopy is performed with a cw, external-cavity spin flip Raman laser containing an intracavity absorption cell. Significantly enhanced sensitivity is demonstrated under marginal oscillator conditions obtained by decreasing the laser gain to a level comparable with the absorption loss of the cell. Spectra of nitric oxide and carbonyl sulfide are discussed to illustrate the performance achieved. PMID- 19680365 TI - Generation of single ultrashort CO(2) laser pulses in a Fabry-Perot interferometer. AB - In a manner analogous to optical free-induction decay, we have generated ultrashort (<100 psec) intense infrared optical pulses by reflecting an abruptly terminated CO(2) laser pulse from an appropriately adjusted Fabry-Perot interferometer. Because a suitable resonantly absorbing material is not required, the scheme is applicable to any intense narrow-band laser system. PMID- 19680366 TI - Dyelike lasers for the 0.9-2-microm region using F(2)+ centers in alkali halides. AB - Optically pumped, cw laser action with F(2)+ centers in the alkali halides is reported. With the host NaF, the laser was continuously tunable over the range 0.885 less, similar lambda less, similar 1.00 microm, and pump power near threshold was ~40 mW at band center. The required high density of F(2)+ centers was created by a simple process involving radiation damage by a 1-MeV e(-) beam. With the F(2)+ in other hosts as well, it should be possible to cover the entire range, 0.9 less, similar lambda less, similar 2 microm. PMID- 19680367 TI - Annular converging wave resonator: new insights. AB - A simple theory describing the annular converging wave resonator has been developed. The theory explains results obtained in two recent experiments. Configurations of the converging wave resonator that could be used to extract power from a large-volume annular gain region in a single mode with nonzero azimuthal order are also discussed. PMID- 19680368 TI - Large-core, broadband optical fiber. AB - A broadband optical fiber with larger core diameter than that of a conventional single-mode fiber is proposed. By appropriately choosing normalized frequency v = 4.6 and core profile parameter alpha = 4.5, a core diameter as large as 16.3 microm with relative index difference Delta = 0.3% at the 1.25-microm wavelength is attainable. Fabrication tolerances, bandwidth, and bending loss of the fiber are discussed. PMID- 19680369 TI - Phase approximation of the Huygens-Kirchhoff method in problems of laser-beam propagation in the turbulent atmosphere. AB - This Letter presents a rigorous mathematical proof of the applicability of phase approximation in the Huygens-Kirchhoff method for the problems of laser-beam propagation in a turbulent atmosphere. The calculated results are in good agreement with experimental data. PMID- 19680370 TI - Single-particle correlated time-of-flight velocimeter for remote wind-speed measurement. AB - A new technique of single-particle correlation for wind-speed measurement by determining aerosol time of flight is discussed. Using this technique, single ended remote measurement of atmospheric wind speeds has been demonstrated at ranges up to 100 m under natural aerosol conditions with less than 0.2-W continuous-wave laser power with a measurement time of approximately 1 sec. PMID- 19680371 TI - Infrared hot carrier diode mixer. AB - Detection of a 54.3-GHz beatnote at 10.6 microm has been observed with a hot carrier diode mixer. The diode consists of a "cat whisker" antenna, which forms an ohmic point contact to n-InAs. The mechanism of this room-temperature detector is described as the "thermoelectric effect" of hot carriers. PMID- 19680373 TI - Measuring the wavelength of light with a self-calibrating grating. PMID- 19680372 TI - Photopolarimeter using two modulated optical rotators. AB - The basic compensator-analyzer-polarimeter is automated by inserting two modulated optical rotators, one in front of the compensator and the other between the compensator and the analyzer. The modulated optical rotators (e.g., two series-connected Faraday cells) are excited by the same ac source to produce sinusoidal optical rotations of the same frequency, generally of different amplitudes, that are mutally in or 180 degrees out of phase. We show that, when the radiation leaving this generalized polarimeter (which is of constant polarization) is linearly detected, limited Fourier analysis of the resulting signal yields the four Stokes parameters of the incident radiation. PMID- 19680374 TI - Influence of higher harmonics of a grating on the intensity profile of the diffraction orders via surface plasmons. PMID- 19680376 TI - Injection-locked, narrow-band KrF discharge laser using an unstable resonator cavity. PMID- 19680375 TI - Mode competition in a ring laser at line center. PMID- 19680377 TI - Complete frequency coverage for submillimeter laser spectroscopy with optically pumped CH3OH, CH3OD, CD3OD, and CH2CF2. PMID- 19680378 TI - Group velocity matching in optical fibers. PMID- 19680380 TI - Multiphoton absorption in the presence of two finite-bandwidth lasers. PMID- 19680379 TI - Optimum photon detection with a simple counting processor. PMID- 19680381 TI - Rotational - vibrational transition moments in excited states of spherical-top molecules. PMID- 19680382 TI - Thermotransmission measurements on self-supporting metal films. PMID- 19680383 TI - Collision-induced rotational Raman scattering. PMID- 19680384 TI - Single-particle light-scattering measurements with a photodiode array. PMID- 19680385 TI - New method for high-range resolution measurements of light scattering in optically dense inhomogeneous media. PMID- 19680387 TI - Correction of phase aberrations via stimulated Brillouin scattering. AB - We have obtained quantitative measurements on the correction of severely aberrated laser beams using stimulated Brillouin scattering (SBS) at 0.69 microm. We have shown that under certain conditions SBS can be used to restore an aberrated optical beam to its original unaberrated condition. When an optical beam double passes an aberrating region after reflecting from an "ordinary" mirror (i.e., a plane mirror) the aberration is twice that obtained from a single pass. However, when the aberrated beam enters a medium that allows SBS to occur, it emerges from its second pass through the aberrating medium in the same condition as that in which it originally entered. Quantitative experiments are described in which a single-mode ruby laser beam is intentionally aberrated by passing it through an etched plate. When the beam is allowed to double-pass the plate using an ordinary reflector (i.e., plane mirror), the beam divergence is more than 10 times the diffraction-limited divergence. However, when we replace the ordinary reflector with a cell in which SBS can take place, the SBS reflected beam is restored to diffraction-limited divergence when it is allowed to pass back through the aberrating medium. Applications of this time-reversal or phase reversal technique for correcting aberrations in optical trains and atmospheric turbulence are discussed. PMID- 19680386 TI - Fully parallel, high-speed incoherent optical method for performing discrete Fourier transforms. AB - An incoherent optical data-processing method is described, which has the potential for performing discrete Fourier transforms of short length at rates far exceeding those afforded by both special-purpose digital hardware and representative coherent optical processors. PMID- 19680388 TI - Thermal blooming compensation with adaptive optics. AB - A multidither adaptive optical system has been used to correct for thermal blooming distortions. Stable correction factors of from 1.3 to 4 have been observed. Turbulence correction in the presence of blooming has also been observed. It is suggested that the geometry of the blooming scenario (location of absorption region, slue rate, etc.) and the parameters of the adaptive system (bandwidth, correction algorithm, etc.) can strongly affect the observed correction factor. PMID- 19680389 TI - Stark-induced optical transients in ruby. AB - Extension of the Brewer-Shoemaker technique of Stark-induced optical transients to a solid-ruby-is reported. Optical free-induction decay and photon echoes, as well as the first observation of an optical edge echo, have been obtained. PMID- 19680390 TI - Lifetime and collisional quenching measurements of XeF*(B) by photolysis of XeF2. AB - By photolyzing XeF 2 using ArCl* 175-nm radiation, the spontaneous radiative lifetime of the XeF(B --> X) band has been measured to be 14.25 + 0.2 nsec. Also, the rates of two-body collisional deactivation of the XeF(B) state by Ne, Ar, and XeF 2 have been determined to be 7.7 X 10(-13) cm3 sec(-1), 4.9 X 10(-12) cm3 sec(-1), and 2.6 X 10(-10) cm3 sec(-1), respectively. A three-body quenching rate of 7.2 X 10(-32) cm6 sec(-1) was found for Ar. The large two- and three-body quenching rates of XeF* by Ar suggest that Ne may be preferable to Ar as a diluent in high-pressure XeF (350-nm) laser mixtures. PMID- 19680391 TI - Intense high-pressure sequence-band CO2 laser. AB - We have obtained temporally smooth, single-line megawatt output from the (0,0 degrees,2) --> [(1,0 degrees,1), (0,2 degrees,1)]1 sequence band in an 1800-Torr CO2 double-discharge laser. These lines were first observed by Reid and Siemsen in a low-pressure discharge at subwatt power outputs [Appl. Phys. Lett. 29, 250 (1976)]. The intense 10-gm region laser transitions reported here may be useful for selective molecular excitation, for nonlinear optics experiments where wavelengths of the usual C02 transitions are inappropriate, and for long-path atmospheric transmission. PMID- 19680392 TI - Adherent dielectric coating of diamond-turned mirrors. AB - Adherent, high-reflectivity infrared, and multilayer dielectric coatings have been applied to diamond-turned aluminum, copper, and electroless nickel substrates. Samples passed cheesecloth, Scotch tape, salt fog, and humidity tests. PMID- 19680393 TI - Hyperbolic cone-channel condenser. AB - An optical condensing system is described that consists of a lens and the polished interior of a cone generated by rotating a hyperbola about an appropriate axis. An optimum concentration of light is achieved in a relatively compact system. In certain circumstances the hyperbola reduces to a straight line, simplifying the construction of the cone. PMID- 19680395 TI - Is complete spatial coherence necessary for the generation of highly directional light beams? PMID- 19680394 TI - Modal caustics and dispersion mechanism in optical fibers. AB - Modal dispersion is explained through the frequency dependence of modal caustics in the ray optics treatment of propagation in multimode optical fibers. Examples are shown for different index profiles that give rise to dispersion equalization or deterioration. PMID- 19680396 TI - Measuring the velocity of individual atoms in real time. PMID- 19680398 TI - Optical collisions in an intense laser field. PMID- 19680397 TI - Efficient phase-matched infrared third-harmonic generation in liquid CO-O2 SF6mixtures. PMID- 19680399 TI - Broadband gas isolator for high-power CO2 lasers. PMID- 19680400 TI - Far-infrared laser system for detection of defects in polyethylene-insulated power cables. PMID- 19680401 TI - Efficient optical waveguide switch/amplitude modulator. PMID- 19680402 TI - Calorimetric measurements of absorption and scattering losses in optical fibers. PMID- 19680403 TI - Image evaluation of the light-enhanced vaporization photolithographic process in As2S3 thin films. PMID- 19680404 TI - Nonlinear electro-optic Fabry-Perot devices using reflected-light feedback. PMID- 19680405 TI - Observation of amplified reflection by degenerate four-wave mixing in atomic sodium vapor. PMID- 19680406 TI - Adaptive laser resonator. PMID- 19680407 TI - Mercuric bromide dissociation laser in an electric discharge. PMID- 19680408 TI - Spectrophone measurements of isotopes of water vapor and nitric oxide and of phosgene at selected wavelengths in the CO and CO 2 laser region. PMID- 19680409 TI - Heterodyne measurements of infrared absorption frequencies of D2O. PMID- 19680410 TI - New continua for absorption spectroscopy from 40 to 2000 A. PMID- 19680411 TI - Analog image transmission in optical waveguides. PMID- 19680412 TI - Contour holography using an injection-locked flash-pumped dye laser. PMID- 19680413 TI - Photopic contrast sensitivity without foveal vision. PMID- 19680415 TI - One-step rainbow hologram. PMID- 19680414 TI - Deep-image rainbow holograms. PMID- 19680416 TI - Quantum analysis of optical bistability and spectrum of fluctuations. PMID- 19680417 TI - High-resolution cw stimulated Raman spectroscopy in molecular hydrogen. PMID- 19680418 TI - Degenerate four-wave mixing in absorbing media. PMID- 19680419 TI - High-J assignments in the 10.5-Mum SF6 spectrum: identification of the levels pumped by CO2 P(12) and P(22). PMID- 19680420 TI - Gain-verification problem in extreme ultraviolet lasing. PMID- 19680421 TI - Half-duplex optical transmission link using an LED source-detector scheme. PMID- 19680422 TI - Pulse broadening in long-span single-mode fibers around a material-dispersion free wavelength. PMID- 19680423 TI - Measuring the Optical Absorption Cross-sections of Au-Ag Nanocages and Au Nanorods by Photoacoustic Imaging. AB - This paper presents a method for measuring the optical absorption cross-sections (sigma(a)) of Au-Ag nanocages and Au nanorods. The method is based on photoacoustic (PA) imaging, where the detected signal is directly proportional to the absorption coefficient (MU(a)) of the nanostructure. For each type of nanostructure, we firstly obtained MU(a) from the PA signal by benchmarking against a linear calibration curve (PA signal vs. MU(a)) derived from a set of methylene blue solutions with different concentrations. We then calculated sigma(a) by dividing the MU(a) by the corresponding concentration of the Au nanostructure. Additonally, we obtained the extinction cross-section (sigma(e), sum of absorption and scattering) from the extinction spectrum recorded using a conventional UV-vis-NIR spectrometer. From the measurements of sigma(a) and sigma(e), we were able to easily derive both the absorption and scattering cross sections for each type of gold nanostructure. The ratios of absorption to extinction obtained from experimental and theoretical approaches agreed well, demonstrating the potential use of this method in determining the optical absorption and scattering properties of gold nanostructures and other types of nanomaterials. PMID- 19680424 TI - Qualia: the geometry of integrated information. AB - According to the integrated information theory, the quantity of consciousness is the amount of integrated information generated by a complex of elements, and the quality of experience is specified by the informational relationships it generates. This paper outlines a framework for characterizing the informational relationships generated by such systems. Qualia space (Q) is a space having an axis for each possible state (activity pattern) of a complex. Within Q, each submechanism specifies a point corresponding to a repertoire of system states. Arrows between repertoires in Q define informational relationships. Together, these arrows specify a quale -- a shape that completely and univocally characterizes the quality of a conscious experience. Phi -- the height of this shape -- is the quantity of consciousness associated with the experience. Entanglement measures how irreducible informational relationships are to their component relationships, specifying concepts and modes. Several corollaries follow from these premises. The quale is determined by both the mechanism and state of the system. Thus, two different systems having identical activity patterns may generate different qualia. Conversely, the same quale may be generated by two systems that differ in both activity and connectivity. Both active and inactive elements specify a quale, but elements that are inactivated do not. Also, the activation of an element affects experience by changing the shape of the quale. The subdivision of experience into modalities and submodalities corresponds to subshapes in Q. In principle, different aspects of experience may be classified as different shapes in Q, and the similarity between experiences reduces to similarities between shapes. Finally, specific qualities, such as the "redness" of red, while generated by a local mechanism, cannot be reduced to it, but require considering the entire quale. Ultimately, the present framework may offer a principled way for translating qualitative properties of experience into mathematics. PMID- 19680426 TI - Nash equilibria in multi-agent motor interactions. AB - Social interactions in classic cognitive games like the ultimatum game or the prisoner's dilemma typically lead to Nash equilibria when multiple competitive decision makers with perfect knowledge select optimal strategies. However, in evolutionary game theory it has been shown that Nash equilibria can also arise as attractors in dynamical systems that can describe, for example, the population dynamics of microorganisms. Similar to such evolutionary dynamics, we find that Nash equilibria arise naturally in motor interactions in which players vie for control and try to minimize effort. When confronted with sensorimotor interaction tasks that correspond to the classical prisoner's dilemma and the rope-pulling game, two-player motor interactions led predominantly to Nash solutions. In contrast, when a single player took both roles, playing the sensorimotor game bimanually, cooperative solutions were found. Our methodology opens up a new avenue for the study of human motor interactions within a game theoretic framework, suggesting that the coupling of motor systems can lead to game theoretic solutions. PMID- 19680425 TI - Temporal controls of the asymmetric cell division cycle in Caulobacter crescentus. AB - The asymmetric cell division cycle of Caulobacter crescentus is orchestrated by an elaborate gene-protein regulatory network, centered on three major control proteins, DnaA, GcrA and CtrA. The regulatory network is cast into a quantitative computational model to investigate in a systematic fashion how these three proteins control the relevant genetic, biochemical and physiological properties of proliferating bacteria. Different controls for both swarmer and stalked cell cycles are represented in the mathematical scheme. The model is validated against observed phenotypes of wild-type cells and relevant mutants, and it predicts the phenotypes of novel mutants and of known mutants under novel experimental conditions. Because the cell cycle control proteins of Caulobacter are conserved across many species of alpha-proteobacteria, the model we are proposing here may be applicable to other genera of importance to agriculture and medicine (e.g., Rhizobium, Brucella). PMID- 19680427 TI - A parsimony approach to biological pathway reconstruction/inference for genomes and metagenomes. AB - A common biological pathway reconstruction approach -- as implemented by many automatic biological pathway services (such as the KAAS and RAST servers) and the functional annotation of metagenomic sequences -- starts with the identification of protein functions or families (e.g., KO families for the KEGG database and the FIG families for the SEED database) in the query sequences, followed by a direct mapping of the identified protein families onto pathways. Given a predicted patchwork of individual biochemical steps, some metric must be applied in deciding what pathways actually exist in the genome or metagenome represented by the sequences. Commonly, and straightforwardly, a complete biological pathway can be identified in a dataset if at least one of the steps associated with the pathway is found. We report, however, that this naive mapping approach leads to an inflated estimate of biological pathways, and thus overestimates the functional diversity of the sample from which the DNA sequences are derived. We developed a parsimony approach, called MinPath (Minimal set of Pathways), for biological pathway reconstructions using protein family predictions, which yields a more conservative, yet more faithful, estimation of the biological pathways for a query dataset. MinPath identified far fewer pathways for the genomes collected in the KEGG database -- as compared to the naive mapping approach -- eliminating some obviously spurious pathway annotations. Results from applying MinPath to several metagenomes indicate that the common methods used for metagenome annotation may significantly overestimate the biological pathways encoded by microbial communities. PMID- 19680428 TI - Wiring neurons with carbon nanotubes. PMID- 19680429 TI - Recognizing sequences of sequences. AB - The brain's decoding of fast sensory streams is currently impossible to emulate, even approximately, with artificial agents. For example, robust speech recognition is relatively easy for humans but exceptionally difficult for artificial speech recognition systems. In this paper, we propose that recognition can be simplified with an internal model of how sensory input is generated, when formulated in a Bayesian framework. We show that a plausible candidate for an internal or generative model is a hierarchy of 'stable heteroclinic channels'. This model describes continuous dynamics in the environment as a hierarchy of sequences, where slower sequences cause faster sequences. Under this model, online recognition corresponds to the dynamic decoding of causal sequences, giving a representation of the environment with predictive power on several timescales. We illustrate the ensuing decoding or recognition scheme using synthetic sequences of syllables, where syllables are sequences of phonemes and phonemes are sequences of sound-wave modulations. By presenting anomalous stimuli, we find that the resulting recognition dynamics disclose inference at multiple time scales and are reminiscent of neuronal dynamics seen in the real brain. PMID- 19680431 TI - A condensation-ordering mechanism in nanoparticle-catalyzed peptide aggregation. AB - Nanoparticles introduced in living cells are capable of strongly promoting the aggregation of peptides and proteins. We use here molecular dynamics simulations to characterise in detail the process by which nanoparticle surfaces catalyse the self-assembly of peptides into fibrillar structures. The simulation of a system of hundreds of peptides over the millisecond timescale enables us to show that the mechanism of aggregation involves a first phase in which small structurally disordered oligomers assemble onto the nanoparticle and a second phase in which they evolve into highly ordered as their size increases. PMID- 19680430 TI - The CUGBP2 splicing factor regulates an ensemble of branchpoints from perimeter binding sites with implications for autoregulation. AB - Alternative pre-mRNA splicing adjusts the transcriptional output of the genome by generating related mRNAs from a single primary transcript, thereby expanding protein diversity. A fundamental unanswered question is how splicing factors achieve specificity in the selection of target substrates despite the recognition of information-poor sequence motifs. The CUGBP2 splicing regulator plays a key role in the brain region-specific silencing of the NI exon of the NMDA R1 receptor. However, the sequence motifs utilized by this factor for specific target exon selection and its role in splicing silencing are not understood. Here, we use chemical modification footprinting to map the contact sites of CUGBP2 to GU-rich motifs closely positioned at the boundaries of the branch sites of the NI exon, and we demonstrate a mechanistic role for this specific arrangement of motifs for the regulation of branchpoint formation. General support for a branch site-perimeter-binding model is indicated by the identification of a group of novel target exons with a similar configuration of motifs that are silenced by CUGBP2. These results reveal an autoregulatory role for CUGBP2 as indicated by its direct interaction with functionally significant RNA motifs surrounding the branch sites upstream of exon 6 of the CUGBP2 transcript itself. The perimeter-binding model explains how CUGBP2 can effectively embrace the branch site region to achieve the specificity needed for the selection of exon targets and the fine-tuning of alternative splicing patterns. PMID- 19680432 TI - Red Queen dynamics with non-standard fitness interactions. AB - Antagonistic coevolution between hosts and parasites can involve rapid fluctuations of genotype frequencies that are known as Red Queen dynamics. Under such dynamics, recombination in the hosts may be advantageous because genetic shuffling can quickly produce disproportionately fit offspring (the Red Queen hypothesis). Previous models investigating these dynamics have assumed rather simple models of genetic interactions between hosts and parasites. Here, we assess the robustness of earlier theoretical predictions about the Red Queen with respect to the underlying host-parasite interactions. To this end, we created large numbers of random interaction matrices, analysed the resulting dynamics through simulation, and ascertained whether recombination was favoured or disfavoured. We observed Red Queen dynamics in many of our simulations provided the interaction matrices exhibited sufficient 'antagonicity'. In agreement with previous studies, strong selection on either hosts or parasites favours selection for increased recombination. However, fast changes in the sign of linkage disequilibrium or epistasis were only infrequently observed and do not appear to be a necessary condition for the Red Queen hypothesis to work. Indeed, recombination was often favoured even though the linkage disequilibrium remained of constant sign throughout the simulations. We conclude that Red Queen-type dynamics involving persistent fluctuations in host and parasite genotype frequencies appear to not be an artefact of specific assumptions about host parasite fitness interactions, but emerge readily with the general interactions studied here. Our results also indicate that although recombination is often favoured, some of the factors previously thought to be important in this process such as linkage disequilibrium fluctuations need to be reassessed when fitness interactions between hosts and parasites are complex. PMID- 19680434 TI - Reference and preference: how does the brain scale subjective value? PMID- 19680433 TI - Effects of attention on what is known and what is not: MEG evidence for functionally discrete memory circuits. AB - Recent results obtained with a neural-network model of the language cortex suggest that the memory circuits developing for words are both distributed and functionally discrete. This model makes testable predictions about brain responses to words and pseudowords under variable availability of attentional resources. In particular, due to their strong internal connections, the action perception circuits for words that the network spontaneously developed exhibit functionally discrete activation dynamics, which are only marginally affected by attentional variations. At the same time, network responses to unfamiliar items - pseudowords - that have not been previously learned (and, therefore, lack corresponding memory representations) exhibit (and predict) strong attention dependence, explained by the different amounts of attentional resources available and, therefore, different degrees of competition between multiple memory circuits partially activated by items lacking lexical traces. We tested these predictions in a novel magnetoencephalography experiment and presented subjects with familiar words and matched unfamiliar pseudowords during attention demanding tasks and under distraction. The magnetic mismatch negativity (MMN) response to words showed relative immunity to attention variations, whereas the MMN to pseudowords exhibited profound variability: when subjects attended the stimuli, the brain response to pseudowords was larger than that to words (as typically observed in the N400); when attention was withdrawn, the opposite pattern emerged, with the response to pseudowords reduced below the response to words. Main cortical sources of these activations were localized to superior-temporal cortex. These results confirm the model's predictions and provide evidence in support of the hypothesis that words are represented in the brain as action-perception circuits that are both discrete and distributed. PMID- 19680435 TI - Shaping what we see: pinning down the influence of value on perceptual judgements. PMID- 19680436 TI - Accelerated immunodeficiency by anti-CCR5 treatment in HIV infection. AB - In 50% of progressing HIV-1 patients, CXCR4-tropic (X4) virus emerges late in infection, often overtaking CCR5-tropic (R5) virus as the dominant viral strain. This "phenotypic switch" is strongly associated with rapidly declining CD4(+) T cell counts and AIDS onset, yet its causes remain unknown. Here, we analyze a mathematical model for the mechanism of X4 emergence in late-stage HIV infection and use this analysis to evaluate the utility of a promising new class of antiretroviral drugs -- CCR5 inhibitors -- in dual R5, X4 infection. The model shows that the R5-to-X4 switch occurs as CD4(+) T cell activation levels increase above a threshold and as CD4(+) T cell counts decrease below a threshold during late-stage HIV infection. Importantly, the model also shows that highly active antiretroviral therapy (HAART) can inhibit X4 emergence but that monotherapy with CCR5 blockers can accelerate X4 onset and immunodeficiency if X4 infection of memory CD4(+) T cells occurs at a high rate. Fortunately, when CXCR4 blockers or HAART are used in conjunction with CCR5 blockers, this risk of accelerated immunodeficiency is eliminated. The results suggest that CCR5 blockers will be more effective when used in combination with CXCR4 blockers and caution against CCR5 blockers in the absence of an effective HAART regimen or during HAART failure. PMID- 19680437 TI - Four distances between pairs of amino acids provide a precise description of their interaction. AB - The three-dimensional structures of proteins are stabilized by the interactions between amino acid residues. Here we report a method where four distances are calculated between any two side chains to provide an exact spatial definition of their bonds. The data were binned into a four-dimensional grid and compared to a random model, from which the preference for specific four-distances was calculated. A clear relation between the quality of the experimental data and the tightness of the distance distribution was observed, with crystal structure data providing far tighter distance distributions than NMR data. Since the four distance data have higher information content than classical bond descriptions, we were able to identify many unique inter-residue features not found previously in proteins. For example, we found that the side chains of Arg, Glu, Val and Leu are not symmetrical in respect to the interactions of their head groups. The described method may be developed into a function, which computationally models accurately protein structures. PMID- 19680438 TI - The Drosophila SH2B family adaptor Lnk acts in parallel to chico in the insulin signaling pathway. AB - Insulin/insulin-like growth factor signaling (IIS) plays a pivotal role in the regulation of growth at the cellular and the organismal level during animal development. Flies with impaired IIS are developmentally delayed and small due to fewer and smaller cells. In the search for new growth-promoting genes, we identified mutations in the gene encoding Lnk, the single fly member of the SH2B family of adaptor molecules. Flies lacking lnk function are viable but severely reduced in size. Furthermore, lnk mutants display phenotypes reminiscent of reduced IIS, such as developmental delay, female sterility, and accumulation of lipids. Genetic epistasis analysis places lnk downstream of the insulin receptor (InR) and upstream of phosphoinositide 3-kinase (PI3K) in the IIS cascade, at the same level as chico (encoding the single fly insulin receptor substrate [IRS] homolog). Both chico and lnk mutant larvae display a similar reduction in IIS activity as judged by the localization of a PIP(3) reporter and the phosphorylation of protein kinase B (PKB). Furthermore, chico; lnk double mutants are synthetically lethal, suggesting that Chico and Lnk fulfill independent but partially redundant functions in the activation of PI3K upon InR stimulation. PMID- 19680439 TI - Why the Indian subcontinent holds the key to global tiger recovery. AB - With only approximately 3,000 wild individuals surviving restricted to just 7% of their historical range, tigers are now a globally threatened species. Therefore, conservation efforts must prioritize regions that harbor more tigers, as well try to capture most of the remaining genetic variation and habitat diversity. Only such prioritization based on demographic, genetic, and ecological considerations can ensure species recovery and retention of evolutionary flexibility in the face of ongoing global changes. Although scientific understanding of ecological and demographic aspects of extant wild tiger populations has improved recently, little is known about their genetic composition and variability. We sampled 73 individual tigers from 28 reserves spread across a diversity of habitats in the Indian subcontinent to obtain 1,263 bp of mitochondrial DNA and 10 microsatellite loci. Our analyses reveals that Indian tigers retain more than half of the extant genetic diversity in the species. Coalescent simulations attribute this high genetic diversity to a historically large population size of about 58,200 tigers for peninsular India south of the Gangetic plains. Furthermore, our analyses indicate a precipitous, possibly human-induced population crash approximately 200 years ago in India, which is in concordance with historical records. Our results suggest that only 1.7% (with an upper limit of 13% and a lower limit of 0.2%) of tiger numbers in historical times remain now. In the global conservation context our results suggest that, based on genetic, demographic, and ecological considerations, the Indian subcontinent holds the key to global survival and recovery of wild tigers. PMID- 19680440 TI - Forests of the night: refugia of genetic diversity in wild tigers. PMID- 19680441 TI - Comparison of the modified fluorescent method and conventional Ziehl-Neelsen method in the detection of acidfast bacilli in lymphnode aspirates. AB - OBJECTIVES: The objectives were to correlate the modified fluorescent method with the conventional Ziehl-Neelsen (ZN) method for the detection of acid-fast bacilli (AFB) and, also to study the efficacy and advantages of using the auramine rhodamine stain on lymph node aspirates under fluorescent microscopy. METHODS: In 108 consecutive patients with a clinical suspicion of tuberculosis (TB) presenting with lymphadenopathy, fine needle aspirations were performed. Smears from the aspirates were processed for routine cytology, the conventional ZN method, and the modified fluorescent method. The significance of the modified fluorescent method over the conventional ZN method was analyzed using the chi square test. RESULTS: Out of 108 aspirates, 102 were studied and remaining 6 were excluded from the study due to diagnosis of malignancy in 4.04% (4/6) and inadequate aspiration in 2.02% (2/6). Among the 102 aspirates, 44.11% (45/102) were positive for AFB on the conventional ZN method, 58.9% (60/102) were indicative of TB on cytology, while the smear positive increased to 81.37% (83/102) on the modified fluorescent method. CONCLUSIONS: Fluorescent microscopy has the advantage of speed and ease of screening, and reduces observer fatigue. The modified fluorescent method was found to be more advantageous than routine cytology and conventional ZN method, particularly in paucibacillary cases. The bacillary positivity rates were higher in the modified fluorescent method than in the ZN method. Hence, the modified fluorescent method can be an adjuvant when used with routine cytology for the identification of AFB. PMID- 19680442 TI - The population and evolutionary dynamics of homologous gene recombination in bacterial populations. AB - In bacteria, recombination is a rare event, not a part of the reproductive process. Nevertheless, recombination -- broadly defined to include the acquisition of genes from external sources, i.e., horizontal gene transfer (HGT) - plays a central role as a source of variation for adaptive evolution in many species of bacteria. Much of niche expansion, resistance to antibiotics and other environmental stresses, virulence, and other characteristics that make bacteria interesting and problematic, is achieved through the expression of genes and genetic elements obtained from other populations of bacteria of the same and different species, as well as from eukaryotes and archaea. While recombination of homologous genes among members of the same species has played a central role in the development of the genetics and molecular biology of bacteria, the contribution of homologous gene recombination (HGR) to bacterial evolution is not at all clear. Also, not so clear are the selective pressures responsible for the evolution and maintenance of transformation, the only bacteria-encoded form of HGR. Using a semi-stochastic simulation of mutation, recombination, and selection within bacterial populations and competition between populations, we explore (1) the contribution of HGR to the rate of adaptive evolution in these populations and (2) the conditions under which HGR will provide a bacterial population a selective advantage over non-recombining or more slowly recombining populations. The results of our simulation indicate that, under broad conditions: (1) HGR occurring at rates in the range anticipated for bacteria like Streptococcus pneumoniae, Escherichia coli, Haemophilus influenzae, and Bacillus subtilis will accelerate the rate at which a population adapts to environmental conditions; (2) once established in a population, selection for this capacity to increase rates of adaptive evolution can maintain bacteria-encoded mechanisms of recombination and prevent invasion of non-recombining populations, even when recombination engenders a modest fitness cost; and (3) because of the density- and frequency dependent nature of HGR in bacteria, this capacity to increase rates of adaptive evolution is not sufficient as a selective force to provide a recombining population a selective advantage when it is rare. Under realistic conditions, homologous gene recombination will increase the rate of adaptive evolution in bacterial populations and, once established, selection for higher rates of evolution will promote the maintenance of bacteria-encoded mechanisms for HGR. On the other hand, increasing rates of adaptive evolution by HGR is unlikely to be the sole or even a dominant selective pressure responsible for the original evolution of transformation. PMID- 19680443 TI - Functional enhancers at the gene-poor 8q24 cancer-linked locus. AB - Multiple discrete regions at 8q24 were recently shown to contain alleles that predispose to many cancers including prostate, breast, and colon. These regions are far from any annotated gene and their biological activities have been unknown. Here we profiled a 5-megabase chromatin segment encompassing all the risk regions for RNA expression, histone modifications, and locations occupied by RNA polymerase II and androgen receptor (AR). This led to the identification of several transcriptional enhancers, which were verified using reporter assays. Two enhancers in one risk region were occupied by AR and responded to androgen treatment; one contained a single nucleotide polymorphism (rs11986220) that resides within a FoxA1 binding site, with the prostate cancer risk allele facilitating both stronger FoxA1 binding and stronger androgen responsiveness. The study reported here exemplifies an approach that may be applied to any risk associated allele in non-protein coding regions as it emerges from genome-wide association studies to better understand the genetic predisposition of complex diseases. PMID- 19680444 TI - Aging and environmental exposures alter tissue-specific DNA methylation dependent upon CpG island context. AB - Epigenetic control of gene transcription is critical for normal human development and cellular differentiation. While alterations of epigenetic marks such as DNA methylation have been linked to cancers and many other human diseases, interindividual epigenetic variations in normal tissues due to aging, environmental factors, or innate susceptibility are poorly characterized. The plasticity, tissue-specific nature, and variability of gene expression are related to epigenomic states that vary across individuals. Thus, population-based investigations are needed to further our understanding of the fundamental dynamics of normal individual epigenomes. We analyzed 217 non-pathologic human tissues from 10 anatomic sites at 1,413 autosomal CpG loci associated with 773 genes to investigate tissue-specific differences in DNA methylation and to discern how aging and exposures contribute to normal variation in methylation. Methylation profile classes derived from unsupervised modeling were significantly associated with age (P<0.0001) and were significant predictors of tissue origin (P<0.0001). In solid tissues (n = 119) we found striking, highly significant CpG island-dependent correlations between age and methylation; loci in CpG islands gained methylation with age, loci not in CpG islands lost methylation with age (P<0.001), and this pattern was consistent across tissues and in an analysis of blood-derived DNA. Our data clearly demonstrate age- and exposure-related differences in tissue-specific methylation and significant age-associated methylation patterns which are CpG island context-dependent. This work provides novel insight into the role of aging and the environment in susceptibility to diseases such as cancer and critically informs the field of epigenomics by providing evidence of epigenetic dysregulation by age-related methylation alterations. Collectively we reveal key issues to consider both in the construction of reference and disease-related epigenomes and in the interpretation of potentially pathologically important alterations. PMID- 19680445 TI - SCA8 CAG/CTG expansions, a tale of two TOXICities: a unique or common case? PMID- 19680446 TI - Multiple Loci within the major histocompatibility complex confer risk of psoriasis. AB - Psoriasis is a common inflammatory skin disease characterized by thickened scaly red plaques. Previously we have performed a genome-wide association study (GWAS) on psoriasis with 1,359 cases and 1,400 controls, which were genotyped for 447,249 SNPs. The most significant finding was for SNP rs12191877, which is in tight linkage disequilibrium with HLA-Cw*0602, the consensus risk allele for psoriasis. However, it is not known whether there are other psoriasis loci within the MHC in addition to HLA-C. In the present study, we searched for additional susceptibility loci within the human leukocyte antigen (HLA) region through in depth analyses of the GWAS data; then, we followed up our findings in an independent Han Chinese 1,139 psoriasis cases and 1,132 controls. Using the phased CEPH dataset as a reference, we imputed the HLA-Cw*0602 in all samples with high accuracy. The association of the imputed HLA-Cw*0602 dosage with disease was much stronger than that of the most significantly associated SNP, rs12191877. Adjusting for HLA-Cw*0602, there were two remaining association signals: one demonstrated by rs2073048 (p = 2 x 10(-6), OR = 0.66), located within c6orf10, a potential downstream effecter of TNF-alpha, and one indicated by rs13437088 (p = 9 x 10(-6), OR = 1.3), located 30 kb centromeric of HLA-B and 16 kb telomeric of MICA. When HLA-Cw*0602, rs2073048, and rs13437088 were all included in a logistic regression model, each of them was significantly associated with disease (p = 3 x 10(-47), 6 x 10(-8), and 3 x 10(-7), respectively). Both putative loci were also significantly associated in the Han Chinese samples after controlling for the imputed HLA-Cw*0602. A detailed analysis of HLA-B in both populations demonstrated that HLA-B*57 was associated with an increased risk of psoriasis and HLA-B*40 a decreased risk, independently of HLA-Cw*0602 and the C6orf10 locus, suggesting the potential pathogenic involvement of HLA-B. These results demonstrate that there are at least two additional loci within the MHC conferring risk of psoriasis. PMID- 19680447 TI - Nuclear calcium signaling controls expression of a large gene pool: identification of a gene program for acquired neuroprotection induced by synaptic activity. AB - Synaptic activity can boost neuroprotection through a mechanism that requires synapse-to-nucleus communication and calcium signals in the cell nucleus. Here we show that in hippocampal neurons nuclear calcium is one of the most potent signals in neuronal gene expression. The induction or repression of 185 neuronal activity-regulated genes is dependent upon nuclear calcium signaling. The nuclear calcium-regulated gene pool contains a genomic program that mediates synaptic activity-induced, acquired neuroprotection. The core set of neuroprotective genes consists of 9 principal components, termed Activity-regulated Inhibitor of Death (AID) genes, and includes Atf3, Btg2, GADD45beta, GADD45gamma, Inhibin beta-A, Interferon activated gene 202B, Npas4, Nr4a1, and Serpinb2, which strongly promote survival of cultured hippocampal neurons. Several AID genes provide neuroprotection through a common process that renders mitochondria more resistant to cellular stress and toxic insults. Stereotaxic delivery of AID gene-expressing recombinant adeno-associated viruses to the hippocampus confers protection in vivo against seizure-induced brain damage. Thus, treatments that enhance nuclear calcium signaling or supplement AID genes represent novel therapies to combat neurodegenerative conditions and neuronal cell loss caused by synaptic dysfunction, which may be accompanied by a deregulation of calcium signal initiation and/or propagation to the cell nucleus. PMID- 19680448 TI - Yersinia enterocolitica targets cells of the innate and adaptive immune system by injection of Yops in a mouse infection model. AB - Yersinia enterocolitica (Ye) evades the immune system of the host by injection of Yersinia outer proteins (Yops) via a type three secretion system into host cells. In this study, a reporter system comprising a YopE-beta-lactamase hybrid protein and a fluorescent staining sensitive to beta-lactamase cleavage was used to track Yop injection in cell culture and in an experimental Ye mouse infection model. Experiments with GD25, GD25-beta1A, and HeLa cells demonstrated that beta1 integrins and RhoGTPases play a role for Yop injection. As demonstrated by infection of splenocyte suspensions in vitro, injection of Yops appears to occur randomly into all types of leukocytes. In contrast, upon infection of mice, Yop injection was detected in 13% of F4/80(+), 11% of CD11c(+), 7% of CD49b(+), 5% of Gr1(+) cells, 2.3% of CD19(+), and 2.6% of CD3(+) cells. Taking the different abundance of these cell types in the spleen into account, the highest total number of Yop-injected cells represents B cells, particularly CD19(+)CD21(+)CD23(+) follicular B cells, followed by neutrophils, dendritic cells, and macrophages, suggesting a distinct cellular tropism of Ye. Yop injected B cells displayed a significantly increased expression of CD69 compared to non-Yop-injected B cells, indicating activation of these cells by Ye. Infection of IFN-gammaR (receptor)- and TNFRp55-deficient mice resulted in increased numbers of Yop-injected spleen cells for yet unknown reasons. The YopE beta-lactamase hybrid protein reporter system provides new insights into the modulation of host cell and immune responses by Ye Yops. PMID- 19680451 TI - Analysis of one-bond Se-Se nuclear couplings in diselenides and 1,2-diselenoles on the basis of molecular orbital theory: torsional angular dependence, electron density influence, and origin in j(Se, Se). AB - Nuclear couplings for the Se-Se bonds, (1)J(Se, Se), are analyzed on the basis of the molecular orbital (MO) theory. The values are calculated by employing the triple zeta basis sets of the Slater type at the DFT level. (1)J(Se, Se) are calculated modeled by MeSeSeMe (1a), which shows the typical torsional angular dependence on varphi(C(Me)SeSeC(Me)). The dependence explains well the observed (1)J(obsd) (Se, Se) of small values ( psi(a) transition, where psi(i) and psi(a) are occupied and unoccupied MO's, respectively. The separate evaluation enables us to recognize and visualize the origin and the mechanism of the couplings. PMID- 19680450 TI - Mycobacterium tuberculosis WhiB3 maintains redox homeostasis by regulating virulence lipid anabolism to modulate macrophage response. AB - The metabolic events associated with maintaining redox homeostasis in Mycobacterium tuberculosis (Mtb) during infection are poorly understood. Here, we discovered a novel redox switching mechanism by which Mtb WhiB3 under defined oxidizing and reducing conditions differentially modulates the assimilation of propionate into the complex virulence polyketides polyacyltrehaloses (PAT), sulfolipids (SL-1), phthiocerol dimycocerosates (PDIM), and the storage lipid triacylglycerol (TAG) that is under control of the DosR/S/T dormancy system. We developed an in vivo radio-labeling technique and demonstrated for the first time the lipid profile changes of Mtb residing in macrophages, and identified WhiB3 as a physiological regulator of virulence lipid anabolism. Importantly, MtbDeltawhiB3 shows enhanced growth on medium containing toxic levels of propionate, thereby implicating WhiB3 in detoxifying excess propionate. Strikingly, the accumulation of reducing equivalents in MtbDeltawhiB3 isolated from macrophages suggests that WhiB3 maintains intracellular redox homeostasis upon infection, and that intrabacterial lipid anabolism functions as a reductant sink. MtbDeltawhiB3 infected macrophages produce higher levels of pro- and anti inflammatory cytokines, indicating that WhiB3-mediated regulation of lipids is required for controlling the innate immune response. Lastly, WhiB3 binds to pks2 and pks3 promoter DNA independent of the presence or redox state of its [4Fe-4S] cluster. Interestingly, reduction of the apo-WhiB3 Cys thiols abolished DNA binding, whereas oxidation stimulated DNA binding. These results confirmed that WhiB3 DNA binding is reversibly regulated by a thiol-disulfide redox switch. These results introduce a new paradigmatic mechanism that describes how WhiB3 facilitates metabolic switching to fatty acids by regulating Mtb lipid anabolism in response to oxido-reductive stress associated with infection, for maintaining redox balance. The link between the WhiB3 virulence pathway and DosR/S/T signaling pathway conceptually advances our understanding of the metabolic adaptation and redox-based signaling events exploited by Mtb to maintain long term persistence. PMID- 19680449 TI - Plasmodium falciparum-mediated induction of human CD25Foxp3 CD4 T cells is independent of direct TCR stimulation and requires IL-2, IL-10 and TGFbeta. AB - CD4(+)CD25(+)Foxp3(+) regulatory T cells (Tregs) regulate disease-associated immunity and excessive inflammatory responses, and numbers of CD4(+)CD25(+)Foxp3(+) Tregs are increased during malaria infection. The mechanisms governing their generation, however, remain to be elucidated. In this study we investigated the role of commonly accepted factors for Foxp3 induction, TCR stimulation and cytokines such as IL-2, TGFbeta and IL-10, in the generation of human CD4(+)CD25(+)Foxp3(+) T cells by the malaria parasite Plasmodium falciparum. Using a co-culture system of malaria-infected red blood cells (iRBCs) and peripheral blood mononuclear cells from healthy individuals, we found that two populations of Foxp3(hi) and Foxp3(int) CD4(+)CD25(hi) T cells with a typical Treg phenotype (CTLA-4(+), CD127(low), CD39(+), ICOS(+), TNFRII(+)) were induced. Pro-inflammatory cytokine production was confined to the Foxp3(int) subset (IFNgamma, IL-4 and IL-17) and inversely correlated with high relative levels of Foxp3(hi) cells, consistent with Foxp3(hi) CD4 T cell-mediated inhibition of parasite-induced effector cytokine T cell responses. Both Foxp3(hi) and Foxp3(int) cells were derived primarily from proliferating CD4(+)CD25(-) T cells with a further significant contribution from CD25(+)Foxp3(+) natural Treg cells to the generation of the Foxp3(hi) subset. Generation of Foxp3(hi), but not Foxp3(int), cells specifically required TGFbeta1 and IL-10. Add-back experiments showed that monocytes expressing increased levels of co-stimulatory molecules were sufficient for iRBC-mediated induction of Foxp3 in CD4 T cells. Foxp3 induction was driven by IL-2 from CD4 T cells stimulated in an MHC class II dependent manner. However, transwell separation experiments showed that direct contact of monocytes with the cells that acquire Foxp3 expression was not required. This novel TCR-independent and therefore antigen-non specific mechanism for by-stander CD4(+)CD25(hi)Foxp3(+) cell induction is likely to reflect a process also occurring in vivo as a consequence of immune activation during malaria infection, and potentially a range of other infectious diseases. PMID- 19680452 TI - The Impact of HIV and Malaria Coinfection: What Is Known and Suggested Venues for Further Study. AB - HIV and malaria have similar global distributions. Annually, 500 million are infected and 1 million die because of malaria. 33 million have HIV and 2 million die from it each year. Minor effects of one infection on the disease course or outcome for the other would significantly impact public health because of the sheer number of people at risk for coinfection. While early population-based studies showed no difference in outcomes between HIV-positive and HIV-negative individuals with malaria, more recent work suggests that those with HIV have more frequent episodes of symptomatic malaria and that malaria increases HIV plasma viral load and decreases CD4+ T cells. HIV and malaria each interact with the host's immune system, resulting in a complex activation of immune cells, and subsequent dysregulated production of cytokines and antibodies. Further investigation of these interactions is needed to better define effects of coinfection. PMID- 19680454 TI - Ultrasonographic study of gallbladder wall thickness and emptying in cirrhotic patients without gallstones. AB - BACKGROUND AND AIM: Gallbladder wall thickening and impaired contractility are currently reported in cirrhotic patients and often related to portal hypertension and hepatic failure. The purpose of this work was to evaluate, by ultrasonographic method, gallbladder wall thickness and gallbladder emptying after a standard meal in normal subjects and in patients with compensated liver cirrhosis without gallstones. METHODS: Twenty-three patients with Child-Pugh class A liver cirrhosis and twenty healthy controls were studied. Gallbladder wall thickness (GWT), gallbladder fasting volume (FV), residual volume (RV), and maximum percentage of emptying (%E) were calculated. Measurements of mean portal velocity, portal vein flow, and serum albumin were performed too. Statistical analysis was assessed by Student's "t test" for unpaired data. RESULTS: GWT was 0.60 +/- 0.22 cm in cirrhotic patients and 0.21 +/- 0.06 cm in controls (P < .0001). FV and RV were, respectively, 37.8 +/- 3.7 cm(3) and 21.8 +/- 3 cm(3) in cirrhotic patients, 21.9 +/- 4.2 cm(3) and 4.6 +/- 2.2 cm(3) in healthy volunteers (P < .0001). %E was smaller in cirrhotics (42.6 +/- 7.8) as compared to controls (80.3 +/- 7.2; P < .0001). CONCLUSIONS: In patients with compensated liver cirrhosis without gallstones gallbladder wall thickness is increased whereas its contractility is reduced. These early structural and functional alterations could play a role in gallstone formation in more advanced stages of the disease. PMID- 19680453 TI - Immunotherapy of head and neck cancer: current and future considerations. AB - Patients with head and neck squamous cell carcinoma (HNSCC) are at considerable risk for death, with 5-year relative survival rates of approximately 60%. The profound multifaceted deficiencies in cell-mediated immunity that persist in most patients after treatment may be related to the high rates of treatment failure and second primary malignancies. Radiotherapy and chemoradiotherapy commonly have severe acute and long-term side effects on immune responses. The development of immunotherapies reflects growing awareness that certain immune system deficiencies specific to HNSCC and some other cancers may contribute to the poor long-term outcomes. Systemic cell-mediated immunotherapy is intended to activate the entire immune system and mount a systemic and/or locoregional antitumor response. The delivery of cytokines, either by single cytokines, for example, interleukin-2, interleukin-12, interferon-gamma, interferon-alpha, or by a biologic mix of multiple cytokines, such as IRX-2, may result in tumor rejection and durable immune responses. Targeted immunotherapy makes use of monoclonal antibodies or vaccines. All immunotherapies for HNSCC except cetuximab remain investigational, but a number of agents whose efficacy and tolerability are promising have entered phase 2 or phase 3 development. PMID- 19680456 TI - Once daily valacyclovir for reducing viral shedding in subjects newly diagnosed with genital herpes. AB - OBJECTIVE: Genital herpes (GH) recurrences and viral shedding are more frequent in the first year after initial HSV-2 infection. The objective of this study was to provide the first evaluation of valacyclovir 1 g once daily compared to placebo in reducing viral shedding in subjects newly diagnosed with GH. METHODS: 70 subjects were randomized to receive valacyclovir 1 g daily or placebo in a crossover design for 60 days with a 7-day washout period. A daily swab of the genital/anal-rectal area was self-collected for HSV-2 detection by PCR. Subjects attended the clinic for routine study visits and GH recurrence visits. Treatment differences were assessed using a nonparametric crossover analysis. RESULTS: 52 subjects had at least one PCR measurement in both treatment periods and comprised the primary efficacy population. Valacyclovir significantly reduced HSV-2 shedding during all days compared to placebo (mean 2.9% versus 13.5% of all days (P < .01), a 78% reduction). Valacyclovir significantly reduced subclinical HSV-2 shedding during all days compared to placebo (mean 2.4% versus 11.0% of all days (P < .01), a 78% reduction). However, 79% of subjects had no GH recurrences while receiving valacyclovir compared to 52% of subjects receiving placebo (P < .01). CONCLUSION: In this study, the frequency of total and subclinical HSV-2 shedding was greater than reported in earlier studies involving subjects with a history of symptomatic genital recurrences. Our study is the first to demonstrate a significant reduction in viral shedding with valacyclovir 1 g daily compared to placebo in a population of subjects newly diagnosed with HSV-2 infection. PMID- 19680457 TI - Mjolnir: extending HAMMER using a diffusion transformation model and histogram equalization for deformable image registration. AB - Image registration is a crucial step in many medical image analysis procedures such as image fusion, surgical planning, segmentation and labeling, and shape comparison in population or longitudinal studies. A new approach to volumetric intersubject deformable image registration is presented. The method, called Mjolnir, is an extension of the highly successful method HAMMER. New image features in order to better localize points of correspondence between the two images are introduced as well as a novel approach to generate a dense displacement field based upon the weighted diffusion of automatically derived feature correspondences. An extensive validation of the algorithm was performed on T1-weighted SPGR MR brain images from the NIREP evaluation database. The results were compared with results generated by HAMMER and are shown to yield significant improvements in cortical alignment as well as reduced computation time. PMID- 19680458 TI - Breast carcinoma cells in primary tumors and effusions have different gene array profiles. AB - The detection of breast carcinoma cells in effusions is associated with rapidly fatal outcome, but these cells are poorly characterized at the molecular level. This study compared the gene array signatures of breast carcinoma cells in primary carcinomas and effusions. The genetic signature of 10 primary tumors and 10 effusions was analyzed using the Array-Ready Oligo set for the Human Genome platform. Results for selected genes were validated using PCR, Western blotting, and immunohistochemistry. Array analysis identified 255 significantly downregulated and 96 upregulated genes in the effusion samples. The majority of differentially expressed genes were part of pathways involved in focal adhesion, extracellular matrix-cell interaction, and the regulation of the actin cytoskeleton. Genes that were upregulated in effusions included KRT8, BCAR1, CLDN4, VIL2, while DCN, CLDN19, ITGA7, and ITGA5 were downregulated at this anatomic site. PCR, Western blotting, and immunohistochemistry confirmed the array findings for BCAR1, CLDN4, VIL2, and DCN. Our data show that breast carcinoma cells in primary carcinomas and effusions have different gene expression signatures, and differentially express a large number of molecules related to adhesion, motility, and metastasis. These differences may have a critical role in designing therapy and in prognostication for patients with metastatic disease localized to the serosal cavities. PMID- 19680455 TI - Regulation of sulfotransferase and UDP-glucuronosyltransferase gene expression by the PPARs. AB - During phase II metabolism, a substrate is rendered more hydrophilic through the covalent attachment of an endogenous molecule. The cytosolic sulfotransferase (SULT) and UDP-glucuronosyltransferase (UGT) families of enzymes account for the majority of phase II metabolism in humans and animals. In general, phase II metabolism is considered to be a detoxication process, as sulfate and glucuronide conjugates are more amenable to excretion and elimination than are the parent substrates. However, certain products of phase II metabolism (e.g., unstable sulfate conjugates) are genotoxic. Members of the nuclear receptor superfamily are particularly important regulators of SULT and UGT gene transcription. In metabolically active tissues, increasing evidence supports a major role for lipid sensing transcription factors, such as peroxisome proliferator-activated receptors (PPARs), in the regulation of rodent and human SULT and UGT gene expression. This review summarizes current information regarding the regulation of these two major classes of phase II metabolizing enzyme by PPARs. PMID- 19680459 TI - Corticotropin-Releasing Hormone (CRH) Programs the Fetal and Maternal Brain. PMID- 19680462 TI - Interaction Site Preference between Carbon Nanotube and Nifedipine: A Combined Density Functional Theory and Classical Molecular Dynamics Study. AB - A novel hybrid density functional theory, MPWB1K, was firstly employed to investigate static adsorptions of a nifedipine on a (10,10) type of single-walled carbon nanotube (SWCNT), which was modeled by C(200)H(40) and C(280) respectively. For both SWCNT models the internal adsorption is more stable than the external adsorption in a range of 5.3-7.8 kcal/mol, which indicates that a nifedipine has a preference to internally adsorb on the (10,10) SWCNT. Molecular dynamic simulations were then used to predict the dynamic behaviors of a nifedipine and the (10, 10) SWCNT system in both gas phase and aqueous solution. The classical MD simulations show that for both cases a nifedipine could spontaneously encapsulate into the SWCNT and migrate in a surprising oscillation behavior inside the SWCNT, however, both phenomena are significantly delayed in the presence of water molecules. The present study suggests that the nanotube network may be used as an efficient tool for transporting this kind of calcium channel antagonists. PMID- 19680463 TI - Optimal sample allocation for design-consistent regression in a cancer services survey when design variables are known for aggregates. AB - We consider optimal sampling rates in element-sampling designs when the anticipated analysis is survey-weighted linear regression and the estimands of interest are linear combinations of regression coefficients from one or more models. Methods are first developed assuming that exact design information is available in the sampling frame and then generalized to situations in which some design variables are available only as aggregates for groups of potential subjects, or from inaccurate or old data. We also consider design for estimation of combinations of coefficients from more than one model. A further generalization allows for flexible combinations of coefficients chosen to improve estimation of one effect while controlling for another. Potential applications include estimation of means for several sets of overlapping domains, or improving estimates for subpopulations such as minority races by disproportionate sampling of geographic areas. In the motivating problem of designing a survey on care received by cancer patients (the CanCORS study), potential design information included block-level census data on race/ethnicity and poverty as well as individual-level data. In one study site, an unequal-probability sampling design using the subjectss residential addresses and census data would have reduced the variance of the estimator of an income effect by 25%, or by 38% if the subjects' races were also known. With flexible weighting of the income contrasts by race, the variance of the estimator would be reduced by 26% using residential addresses alone and by 52% using addresses and races. Our methods would be useful in studies in which geographic oversampling by race-ethnicity or socioeconomic characteristics is considered, or in any study in which characteristics available in sampling frames are measured with error. PMID- 19680460 TI - Antiviral activity of innate immune protein ISG15. AB - The host innate immune response, including the production of type-I IFN, represents the primary line of defense against invading viral pathogens. Of the hundreds of IFN-stimulated genes (ISGs) discovered to date, ISG15 was one of the first identified and shown to encode a ubiquitin-like protein that functions, in part, as a modifier of protein function. Evidence implicating ISG15 as an innate immune protein with broad-spectrum antiviral activity continues to accumulate rapidly. This review will summarize recent findings on the innate antiviral activity of ISG15, with a focus on the interplay between ubiquitination and ISGylation pathways resulting in modulation of RNA virus assembly/budding. Indeed, ubiquitination is known to be proviral for some RNA viruses, whereas the parallel ISGylation pathway is known to be antiviral. A better understanding of the antiviral activities of ISG15 will enhance our fundamental knowledge of host innate responses to viral pathogens and may provide insight useful for the development of novel therapeutic approaches designed to enhance the immune response against such pathogens. PMID- 19680464 TI - Production and processing of siRNA precursor transcripts from the highly repetitive maize genome. AB - Mutations affecting the maintenance of heritable epigenetic states in maize identify multiple RNA-directed DNA methylation (RdDM) factors including RMR1, a novel member of a plant-specific clade of Snf2-related proteins. Here we show that RMR1 is necessary for the accumulation of a majority of 24 nt small RNAs, including those derived from Long-Terminal Repeat (LTR) retrotransposons, the most common repetitive feature in the maize genome. A genetic analysis of DNA transposon repression indicates that RMR1 acts upstream of the RNA-dependent RNA polymerase, RDR2 (MOP1). Surprisingly, we show that non-polyadenylated transcripts from a sampling of LTR retrotransposons are lost in both rmr1 and rdr2 mutants. In contrast, plants deficient for RNA Polymerase IV (Pol IV) function show an increase in polyadenylated LTR RNA transcripts. These findings support a model in which Pol IV functions independently of the small RNA accumulation facilitated by RMR1 and RDR2 and support that a loss of Pol IV leads to RNA Polymerase II-based transcription. Additionally, the lack of changes in general genome homeostasis in rmr1 mutants, despite the global loss of 24 nt small RNAs, challenges the perceived roles of siRNAs in maintaining functional heterochromatin in the genomes of outcrossing grass species. PMID- 19680465 TI - Routine review of ascites fluid from patients with cirrhosis or hepatocellular carcinoma is a low-yield procedure: an observational study. AB - BACKGROUND: Patients with cirrhosis develop ascites for physiologic reasons that are unrelated to malignant progression. However, physicians performing paracentesis in these patients, often send fluid to the cytology laboratory, sometimes specifically looking for hepatocellular carcinoma (HCC). We have investigated the diagnostic yield of these specimens. MATERIALS AND METHODS: A computerized pathology database search for all ascites fluid cases submitted to the cytology laboratory at a major liver transplant center between November 2004 and April 2008 was performed. Clinical history was obtained for each case. Patients with cirrhosis, with or without HCC, were included in the study. Cytologic diagnoses were compiled and follow-up information was obtained for cases with non-negative findings. RESULTS: A total of 167 specimens from 133 patients ranging from 29 to 85 years of age (mean 56 years) were submitted over the said time period. The causes of cirrhosis included: alcohol - 44; Hepatitis C - 30; Hepatitis B - 6; non-alcoholic steatohepatitis - 7; cryptogenic - 18; other single causes - 6; and multifactorial (alcohol and hepatitis viruses) - 22. Hepatocellular carcinoma (HCC) was present or strongly suspected in 17 patients and had been previously resected in two others. The status of fifteen patients was post liver transplant, with recurrent liver failure. Human immunodeficiency virus was present in seven patients and eight patients had a history of non hepatic malignancies. Among the specimens, 162 were negative, two had atypical lymphocytes worked up for lymphoma, and three had atypical epithelioid cells; none was positive for HCC. Immunohistochemistry demonstrated a mesothelial origin for the atypical epithelioid cells in two cases; in the third case, the patient died shortly after the specimen was collected, with no radiological evidence of HCC. CONCLUSION: Ascites fluid cytology specimens in patients with cirrhosis, even those known or suspected to have HCC, are almost always negative. Atypical cells seen in such specimens should be treated with skepticism since the likelihood that they represent peritoneal spread of HCC is low.. PMID- 19680466 TI - Posterior midline and ventral parietal activity is associated with retrieval success and encoding failure. AB - The ventral part of lateral posterior parietal cortex (VPC) and the posterior midline region (PMR), including the posterior cingulate cortex and precuneus, tend to show deactivation during demanding cognitive tasks, and have been associated with the default mode of the brain. Interestingly, PMR and VPC activity has been associated with successful episodic retrieval but also with unsuccessful episodic encoding. However, the differential contributions of PMR and VPC to retrieval vs. encoding has never been demonstrated within-subjects and within the same experiment. Here, we directly tested the prediction that PMR and VPC activity should be associated with retrieval success but with encoding failure. Consistent with this prediction, we found across five different fMRI experiments that, during retrieval, activity in these regions is greater for hits than misses, whereas during encoding, it is greater for subsequent misses than hits. We also found that these regions overlap with the ones that show deactivations during conscious rest. Our findings further aid in clarifying the role of the default mode regions in learning and memory. PMID- 19680467 TI - Form-function relations in cone-tipped stimulating microelectrodes. AB - Metal microelectrodes are widely used in neuroscience research, and could potentially replace macroelectrodes in various neuro-stimulation applications where their small size, specificity, and their ability to also measure unit activity are desirable. The design of stimulating microelectrodes for specific applications requires knowledge on how tip geometry affects function, but several fundamental aspects of this relationship are not yet well understood. This study uses a combined experimental and physical finite elements simulation approach to formulate three new relationships between the geometrical and electrical properties of stimulating cone-tipped tungsten microelectrodes: (1) The empirical relationship between microelectrode 1-kHz impedance and the exposed tip surface area is best approximated by an inverse square-root function (as expected for a cone-tipped resistive interface). (2) Tip angle plays a major role in determining current distribution along the tip, and as a consequence crucially affects the charge injection capacity of a microelectrode. (3) The critical current for the onset of corrosion is independent of tip surface area in sharp microelectrodes. PMID- 19680468 TI - A model for cortical rewiring following deafferentation and focal stroke. AB - It is still unclear to what extent structural plasticity in terms of synaptic rewiring is the cause for cortical remapping after a lesion. Recent two-photon laser imaging studies demonstrate that synaptic rewiring is persistent in the adult brain and is dramatically increased following brain lesions or after a loss of sensory input (cortical deafferentation). We use a recurrent neural network model to study the time course of synaptic rewiring following a peripheral lesion. For this, we represent axonal and dendritic elements of cortical neurons to model synapse formation, pruning and synaptic rewiring. Neurons increase and decrease the number of axonal and dendritic elements in an activity-dependent fashion in order to maintain their activity in a homeostatic equilibrium. In this study we demonstrate that synaptic rewiring contributes to neuronal homeostasis during normal development as well as following lesions. We show that networks in homeostasis, which can therefore be considered as adult networks, are much less able to compensate for a loss of input. Interestingly, we found that paused stimulation of the networks are much more effective promoting reorganization than continuous stimulation. This can be explained as neurons quickly adapt to this stimulation whereas pauses prevents a saturation of the positive stimulation effect. These findings may suggest strategies for improving therapies in neurologic rehabilitation. PMID- 19680470 TI - Perfusion imaging of the right perisylvian neural network in acute spatial neglect. AB - Recent studies have suggested a tightly connected perisylvian neural network associated with spatial neglect. Here we investigated whether structural damage in one part of the network typically is accompanied with functional damage in other, structurally intact areas of this network. By combining normalized fluid attenuated inversion-recovery (FLAIR) imaging, diffusion-weighted imaging (DWI), and perfusion-weighted imaging (PWI) we asked whether or not lesions centering on fronto-temporal regions co-occur with abnormal perfusion in structurally intact parietal cortex. With thresholds applied to delineate behaviourally relevant malperfusion of brain tissue, the analysis of normalized time-to-peak (TTP) and maximal signal reduction (MSR) perfusion maps did not reveal significant changes outside the area of structural damage. In particular, we found no abnormal perfusion in the structurally intact inferior parietal lobule (IPL) and/or the temporo-parietal junction (TPJ). The present results obtained in three consecutively admitted neglect patients with fronto-temporal lesions indicate that structural damage in one part of the right perisylvian network associated with spatial neglect does not necessarily require dysfunction by malperfusion in other, structurally intact parts of the network to provoke spatial neglect. The neural tissue in the fronto-temporal cortex appears to have an original role in processes of spatial orienting and exploration. PMID- 19680469 TI - Enhancing m currents: a way out for neuropathic pain? AB - Almost three decades ago, the M current was identified and characterized in frog sympathetic neurons (Brown and Adams, 1980). The years following this discovery have seen a huge progress in the understanding of the function and the pharmacology of this current as well as on the structure of the underlying ion channels. Therapies for a number of syndromes involving abnormal levels of excitability in neurons are benefiting from research on M currents. At present, the potential of M current openers as analgesics for neuropathic pain is under discussion. Here we offer a critical view of existing data on the involvement of M currents in pain processing. We believe that enhancement of M currents at the site of injury may become a powerful strategy to alleviate pain in some peripheral neuropathies. PMID- 19680471 TI - An apoA-I mimetic peptide increases LCAT activity in mice through increasing HDL concentration. AB - Lecithin cholesterol acyltransferase (LCAT) plays a key role in the reverse cholesterol transport (RCT) process by converting cholesterol to cholesteryl ester to form mature HDL particles, which in turn deliver cholesterol back to the liver for excretion and catabolism. HDL levels in human plasma are negatively correlated with cardiovascular risk and HDL functions are believed to be more important in atheroprotection. This study investigates whether and how D-4F, an apolipoprotein A-I (apoA-I) mimetic peptide, influences LCAT activity in the completion of the RCT process. We demonstrated that the apparent rate constant value of the LCAT enzyme reaction gives a measure of LCAT activity and determined the effects of free metals and a reducing agent on LCAT activity, showing an inhibition hierarchy of Zn(2+)>Mg(2+)>Ca(2+) and no inhibition with beta mercaptoethanol up to 10 mM. We reconstituted nano-disc particles using apoA-I or D-4F with phospholipids. These particles elicited good activity in vitro in the stimulation of cholesterol efflux from macrophages through the ATP-binding cassette transporter A1 (ABCA1). With these particles we studied the LCAT activity and demonstrated that D-4F did not activate LCAT in vitro. Furthermore, we have done in vivo experiments with apoE-null mice and demonstrated that D-4F (20 mg/kg body weight, once daily subcutaneously) increased LCAT activity and HDL level as well as apoA-I concentration at 72 hours post initial dosing. Finally, we have established a correlation between HDL concentration and LCAT activity in the D-4F treated mice. PMID- 19680473 TI - A pilot study of rizatriptan and visually-induced motion sickness in migraineurs. AB - BACKGROUND: Limited evidence suggests that rizatriptan given before vestibular stimulation reduces motion sickness in persons with migraine-related dizziness. The present study was designed to test whether rizatriptan is also effective in protecting against visually-induced motion sickness and to test whether rizatriptan blocks the augmentation of motion sickness by head pain. MATERIAL AND METHODS: Using randomized double-blind, placebo-controlled methodology, 10 females, 6 with migrainous vertigo (V+) and four without vertigo (V-) received 10 mg rizatriptan or placebo two hours prior to being stimulated by optokinetic stripes. Visual stimulation was coupled with three pain conditions: no pain (N), thermally-induced hand pain (H) and temple pain (T). Motion sickness and subjective discomfort were measured. RESULTS: Motion sickness was less after pre treatment with rizatriptan for 4 of 10 subjects and more for 5 of 10 subjects. Augmentation of motion sickness by head pain was seen in 6 of 10 subjects; this effect was blunted by rizatriptan in 4 of these 6 subjects. Subjective discomfort was significantly more noticeable in V+ subjects as compared with V- subjects. CONCLUSIONS: These pilot data suggest that rizatriptan does not consistently reduce visually-induced motion sickness in migraineurs. Rizatriptan may diminish motion sickness potentiation by cranial pain. PMID- 19680472 TI - The prospects of cellulase-producing bacteria for the bioconversion of lignocellulosic biomass. AB - Lignocellulosic biomass is a renewable and abundant resource with great potential for bioconversion to value-added bioproducts. However, the biorefining process remains economically unfeasible due to a lack of biocatalysts that can overcome costly hurdles such as cooling from high temperature, pumping of oxygen/stirring, and, neutralization from acidic or basic pH. The extreme environmental resistance of bacteria permits screening and isolation of novel cellulases to help overcome these challenges. Rapid, efficient cellulase screening techniques, using cellulase assays and metagenomic libraries, are a must. Rare cellulases with activities on soluble and crystalline cellulose have been isolated from strains of Paenibacillus and Bacillus and shown to have high thermostability and/or activity over a wide pH spectrum. While novel cellulases from strains like Cellulomonas flavigena and Terendinibacter turnerae, produce multifunctional cellulases with broader substrate utilization. These enzymes offer a framework for enhancement of cellulases including: specific activity, thermalstability, or end-product inhibition. In addition, anaerobic bacteria like the clostridia offer potential due to species capable of producing compound multienzyme complexes called cellulosomes. Cellulosomes provide synergy and close proximity of enzymes to substrate, increasing activity towards crystalline cellulose. This has lead to the construction of designer cellulosomes enhanced for specific substrate activity. Furthermore, cellulosome-producing Clostridium thermocellum and its ability to ferment sugars to ethanol; its amenability to co-culture and, recent advances in genetic engineering, offer a promising future in biofuels. The exploitation of bacteria in the search for improved enzymes or strategies provides a means to upgrade feasibility for lignocellulosic biomass conversion, ultimately providing means to a 'greener' technology. PMID- 19680474 TI - Effect of corticosteroids on phlebitis induced by intravenous infusion of antineoplastic agents in rabbits. AB - PURPOSE: Phlebitis caused by intravenous infusion of antineoplastic agents is one of the critical problems when anticancer therapy is prolonged. We have already reported that both rapid infusion and dilution of the injection solution were effective methods for reducing phlebitis caused by vinorelbine (VNR) in rabbits. The aim of this study was to explore other practical methods for preventing phlebitis caused by VNR and doxorubicin (DXR) in a rabbit model. VNR is often used with cisplatin, and dexamethasone (DEX) has been co-administered for prevention of cisplatin-induced nausea. DXR is used with prednisolone (PSL) in the CHOP regimen for the treatment of non-Hodgkin's lymphoma. Therefore, the present study investigated the prevention of phlebitis due to VNR with DEX and that due to DXR with PSL. METHODS: VNR and DXR were diluted with normal saline to prepare test solutions at concentrations of 0.6 mg/mL and 1.4 mg/mL, respectively. Each test solution was infused into the auricular veins of rabbits. Two days after VNR infusion and three days after DXR infusion, the veins were evaluated histopathologically. The effect of DEX on VNR-induced phlebitis was evaluated by infusion of DEX before or after VNR. The effect of PSL on DXR induced phlebitis was similarly evaluated by co-infusion of PSL. RESULTS: The histopathological features of phlebitis caused by the antineoplastic agents differed between VNR and DXR: VNR did not cause the loss of venous endothelial cells, but caused inflammatory cell infiltration, edema, and epidermal degeneration. In contrast, DXR caused the loss of venous endothelial cells and chrondrocyte necrosis. Pre-treatment and post-treatment with DEX significantly decreased VNR-induced phlebitis compared with the control group and pre-treatment was particularly effective. Co-infusion of PSL also significantly decreased phlebitis caused by DXR, but its effect was less marked. CONCLUSION: The present findings suggested that pre-treatment with DEX may be a useful method for preventing phlebitis due to VNR, and that co-infusion of PSL has the potential to prevent phlebitis caused by DXR. PMID- 19680475 TI - Gene expression profiling of chromophobe renal cell carcinomas and renal oncocytomas by Affymetrix GeneChip using pooled and individual tumours. AB - Due to overlapping morphology, malignant chromophobe renal cell carcinomas (RCC) and benign renal oncocytomas (RO) may pose a diagnostic problem. In the present study, we have applied different algorithms to evaluate the data sets obtained by hybridisation of pooled and also individual samples of renal cell tumours (RCT) onto two different gene expression platforms. The two approaches revealed high similarities in the gene expression profiles of chromophobe RCCs and ROs but also some differences. After identifying the differentially expressed genes by statistic analyses, the candidate genes were further selected by a real time and normal RT-PCR and their products were analysed by immunohistochemistry. We have identified CD82 and S100A1 as valuable markers for chromophobe RCC as well as AQP6 for ROs. However, these genes are expressed at the protein level in other types of RCTs as well albeit at a low frequency and low intensity. As none of the selected genes marks exclusively one type of RCTs, for the differential diagnosis of chromophobe RCCs and ROs, a set of markers such as CD82, S100A1 and AQP6 as well as some others would be an option in routine histological laboratories. PMID- 19680478 TI - Rights to individual privacy and professional confidentiality - A Canadian Paediatric Surveillance Program ongoing commitment. PMID- 19680476 TI - A novel peptide inhibits the influenza virus replication by preventing the viral attachment to the host cells. AB - Avian influenza viruses (AIV), the causative agent of avian flu or bird flu, cause widespread morbidity and mortality in poultry. The symptoms of the disease range from mild flu like symptoms to death. These viruses possess two important surface glycoproteins, namely hemagglutinin (HA) and neuraminidase (NA) against which neutralizing antibodies are produced. Due to the highly mutative nature of the genes which encode these proteins, the viruses often confer resistance to the current anti-viral drugs making the prevention and treatment of infection challenging. In our laboratory, we have recently identified a novel anti-viral peptide (P1) against the AIV H9N2 from a phage displayed peptide library. This peptide inhibits the replication of the virus in ovo and in vitro by its binding to the HA glycoprotein. In the current study, we demonstrate that the peptide inhibits the virus replication by preventing the attachment to the host cell but it does not have any effect on the viral fusion. The reduction in the viral nucleoprotein (NP) expression inside the host cell has also been observed during the peptide (P1) treatment. This novel peptide may have the potential to be developed as a therapeutic agent for the treatment and control of avian influenza virus H9N2 infections. PMID- 19680477 TI - Improving access to mental health services for youth and parents. PMID- 19680479 TI - The aggressive and impulsive child: Innovations in assessment and treatment - A commentary. PMID- 19680480 TI - Antidepressant use in adolescence: We're asking the wrong questions. PMID- 19680481 TI - Infant-parent attachment: Definition, types, antecedents, measurement and outcome. AB - Attachment theory is one of the most popular and empirically grounded theories relating to parenting. The purpose of the present article is to review some pertinent aspects of attachment theory and findings from attachment research. Attachment is one specific aspect of the relationship between a child and a parent with its purpose being to make a child safe, secure and protected. Attachment is distinguished from other aspects of parenting, such as disciplining, entertaining and teaching. Common misconceptions about what attachment is and what it is not are discussed. The distinction between attachment and bonding is provided. The recognized method to assess infant-parent attachment, the Strange Situation procedure, is described. In addition, a description is provided for the four major types of infant-parent attachment, ie, secure, insecure-avoidant, insecure-resistant and insecure-disorganized. The antecedents and consequences of each of the four types of infant-parent attachment are discussed. A special emphasis is placed on the description of disorganized attachment because of its association with significant emotional and behavioural problems, and poor social and emotional outcomes in high-risk groups and in the majority of children who have disorganized attachment with their primary caregiver. Practical applications of attachment theory and research are presented. PMID- 19680482 TI - Behavioural inhibition: A predictor of anxiety. AB - Anxiety disorders are prevalent and contribute to emotional suffering and significant economic loss. Early identification and treatment are essential, not only to reduce the associated morbidity, disability and mortality of the anxiety disorders themselves, but also to minimize development of frequent comorbidities such as depression and substance abuse. To understand the factors that increase susceptibility to developing anxiety disorders, a temperamental construct called behavioural inhibition, which refers to the consistent tendency of some children to demonstrate fear and withdrawal in novel situations, has been developed. The present article reviews studies investigating this model as a premorbid predictor of those at risk for developing anxiety disorders, including prospective studies of children at risk as well as retrospective and family studies. In summary, these data suggest the usefulness of this model and a need for further research to determine the optimal management of behaviourally inhibited children as a potential way to prevent adult psychopathology. PMID- 19680483 TI - Adolescent-parent attachment: Bonds that support healthy development. AB - Adolescence is characterized by significant neurological, cognitive and sociopsychological development. With the advance of adolescence, the amount of time spent with parents typically drops while time spent with peers increases considerably. Nonetheless, parents continue to play a key role in influencing their adolescent's development. Adolescent-parent attachment has profound effects on cognitive, social and emotional functioning. Secure attachment is associated with less engagement in high risk behaviours, fewer mental health problems, and enhanced social skills and coping strategies. The present article provides a brief synopsis of the changes that occur during adolescence and describes what attachment is, why it continues to be important and how it is transformed during adolescence. It summarizes major findings on the impact of attachment on adolescent adjustment and discusses strategies for supporting healthy adolescent parent attachment. PMID- 19680484 TI - Erb's palsy - Who is to blame and what will happen? AB - Erb's palsy is initially frightening. The infant's arm hangs limply from the shoulder with flexion of the wrist and fingers due to weakness of muscles innervated by cervical roots C5 and C6. Risk factors are macrosomia (large baby) and shoulder dystocia. However, Erb's palsy may occur following cesarian section. The experience of the delivering physician may not influence the risk of Erb's palsy (0.9 to 2.6 per 1000 live births). Differential diagnosis includes clavicular fracture, osteomyelitis and septic arthritis. Fortunately, the rate of complete recovery is 80% to 96%, especially if improvement begins in the first two weeks. Recommended treatment includes early immobilization followed by passive and active range of motion exercises (although there is no proof that any intervention is effective). For the few infants with no recovery by three to five months, surgical exploration of the brachial plexus may improve the outcome. Three infants with Erb's palsy who illustrate variations in the evolution of this disorder are presented. PMID- 19680486 TI - Varicella (chickenpox). PMID- 19680485 TI - Adolescent parents and their children - The paediatrician's role. AB - Adolescent parents and their children have a number of health issues that require attention from the health care provider. Both the adolescent and their child have health needs, and the young parent often requires significant support in their parenting role. Paediatricians should be aware of the areas in which problems can develop, including maternal nutrition and emotional health, and infant growth and development. The literature addressing these areas of concern will be reviewed and general recommendations for the care of this patient population will be provided. PMID- 19680487 TI - Prescription drug costs. PMID- 19680488 TI - In preterm infants, does the supplementation of carnitine to parenteral nutrition improve the following clinical outcomes: Growth, lipid metabolism and apneic spells?: Part A: Evidence-based answer and summary. PMID- 19680489 TI - In preterm infants, does the supplementation of carnitine to parenteral nutrition improve the following clinical outcomes: Growth, lipid metabolism and apneic spells?: Part B: Clinical commentary. PMID- 19680491 TI - Depression in pregnant women and mothers: How children are affected. PMID- 19680492 TI - Extra-axial medulloblastoma in the cerebellopontine angle. PMID- 19680493 TI - Intestinal duplication presenting with recurrent abdominal pain. PMID- 19680490 TI - Maternal depression and child development. PMID- 19680494 TI - Lymphatic cording or axillary web syndrome after breast cancer surgery. PMID- 19680495 TI - Malakoplakia of the neck in a patient with cancer of the colon. PMID- 19680496 TI - Massive hemobilia. PMID- 19680497 TI - Nodular ganglioneuroblastoma in adults. PMID- 19680498 TI - Unexpected bile duct stones formed around surgical clips 4 years after laparoscopic cholecystectomy. PMID- 19680499 TI - Cerebellar liponeurocytoma. PMID- 19680500 TI - Spontaneous intrahepatic echinococcal cyst rupture in a patient with chronic hepatitis C infection. PMID- 19680501 TI - Lymphoma following ileal pouch anal anastomosis. PMID- 19680502 TI - Influence of delays to nonemergent colon cancer surgery on operative mortality, disease-specific survival and overall survival. AB - BACKGROUND: There has been minimal research on the influence of delays for cancer treatments on patient outcomes. We measured the influence of delays to nonemergent colon cancer surgery on operative mortality, disease-specific survival and overall survival. METHODS: We used the linked Surveillance, Epidemiology and End Results (SEER)-Medicare databases (1993-1996) to identify patients who underwent nonemergent colon cancer surgery. We assessed 2 time intervals: surgeon consult to hospital admission for surgery and first diagnostic test for colon cancer to hospital admission. Follow-up data were available to the end of 2003. We selected the time intervals to create patient groups with clinical relevance and they did not extend past 120 days. RESULTS: We identified 7989 patients who underwent nonemergent colon cancer surgery. Median delays from surgeon consult to admission and from first diagnostic test to admission were 7 and 17 days, respectively. The odds of operative mortality were similar if the consult-to-admission interval was 22 days or more versus 1-7 days (odds ratio [OR] 1.0, 95% confidence interval [CI] 0.6-1.8, p = 0.91) or if the test-to admission interval was 43 days or more versus 1-14 days (OR 0.8, 95% CI 0.4-1.5, p = 0.51), respectively. For these same respective interval comparisons, disease specific survival was not influenced by the consult-to-admission wait (hazard ratio [HR] 1.0, 95% CI 0.9-1.2, p = 0.91) or the test-to-admission wait (HR 1.0, 95% CI 0.8-1.1, p = 0.63). The risk of death was slightly greater if the consult to-admission interval was 22 or more days versus 1-7 days (HR 1.1, 95% CI 1.0 1.2, p = 0.013) and if the test-to-admission interval was 43 days or more versus 1-14 days (HR 1.2, 95% CI 1.1-1.3, p = 0.003). CONCLUSION: It is unlikely that delays to nonemergent colon cancer surgery longer than 3 weeks from initial surgical consult or longer than 6 weeks from first diagnostic test negatively impact operative mortality, disease-specific survival or overall survival. PMID- 19680503 TI - Seven-year survival after intralesional resection and adjuvant radiotherapy for a giant-cell tumour of the sixth cervical vertebra. PMID- 19680504 TI - Subtotal splenectomy for splenic abscess. PMID- 19680505 TI - Primary myxoid sarcoma of the pleura. PMID- 19680506 TI - An unusual case of free air. PMID- 19680507 TI - Paradoxical carbon dioxide embolism during laparoscopic unroofing of a recurrent nonparasitic liver cyst. PMID- 19680508 TI - Ruptured ectopic pregnancy presenting as intestinal obstruction. PMID- 19680509 TI - Toward a platform for training and practice standards in advanced minimally invasive surgery. PMID- 19680510 TI - Predictors of laparotomy and mortality in polytrauma patients with pelvic fractures. AB - BACKGROUND: The decision to perform laparotomy in blunt trauma patients is often difficult owing to pelvic fractures; however, once the decision is made, delay or failure to perform laparotomy could affect morbidity and mortality. We sought to identify predictors of laparotomy and mortality in polytrauma patients with pelvic fractures. METHODS: We divided 390 blunt polytrauma patients (Injury Severity Score [ISS] >/= 16) with pelvic fractures into laparotomy (n = 56) and nonlaparotomy (n = 334) groups. We assessed the role of the following variables in predicting laparotomy and mortality: age, sex, hypotension, fluid and blood transfusions, positive abdominal computed tomography (CT) scans or focused assessment with sonography for trauma (FAST) examination, pelvic fracture severity and ISS. We analyzed the data using Student t and chi(2) tests, followed by logistic regression analysis. RESULTS: Mortality was higher in the laparotomy group than the nonlaparotomy group (28.6% v. 12.9%; overall mortality 15.1%). The laparotomy group had higher mean ISS (36.9 v. 24.9), higher mean abbreviated injury scores (AIS) for the abdomen (2.6 v. 0.9) and chest (3.4 v. 1.6), lower mean initial hemoglobin levels (105.2 v. 127.0 g/L), higher mean crystalloid (4249 v. 3436 mL) and blood transfusion volumes over 4 hours (12.1 v. 3.9 units), more frequent hypotension (44.6 v. 18.0%) and a higher percentage of positive CT scans (67.9% v. 28.4%) and FAST examination results (42.9% v. 3.3%) than the nonlaparotomy group. Age (mean 53.7 v. 41.5 yr); ISS (mean 39.0 v. 24.4); AIS for the head (mean 3.2 v. 1.7), abdomen (mean 1.6 v. 1.1), chest (mean 2.7 v. 1.8) and pelvis (mean 3.1 v. 2.6); crystalloid (mean 5157.3 v. 3266.4 mL) and blood transfusion volumes over 4 hours (mean 13.1 v. 3.7) and initial hypotension (61% v. 14.8%) were all greater among patients who died than those who survived. Mean initial hemoglobin levels were lower among patients who died than among those who survived (111.1 v. 126.2 g/L). Age, the AIS for the head, initial hypotension and low initial hemoglobin levels were highly predictive of mortality, whereas low initial hemoglobin levels, a positive FAST examination and high AIS for the abdomen and chest were all highly predictive of laparotomy. CONCLUSION: Among the polytrauma patients with pelvic fractures, 14.3% underwent laparotomy, and mortality was higher among these patients than among those who did not have the procedure. The predictors of laparotomy and mortality are similar to those anticipated in patients without pelvic fractures. PMID- 19680511 TI - The interobserver reliability of classification systems for radial head fractures: the Hotchkiss modification of the Mason classification and the AO classification systems. AB - BACKGROUND: Radial head fractures are common injuries, and there is little information on the reliability of classification systems for such injuries. The purpose of our study was to report the interobserver reliability of 2 commonly used classification systems: the Hotchkiss modification of the Mason classification and the AO classification systems. METHODS: We compiled the radiographs from a cohort series of 43 patients with radial head fractures, and 5 observers classified the radiographs according to both classification systems. Additionally, we collapsed the systems, with types II and III combined for the Hotchkiss classification and the final digit dropped for the AO classification. We calculated percent agreement, the kappa statistic and the associated 95% confidence intervals (CIs). RESULTS: The mean percent agreement was 72.3% (95% CI 65.8%-78.9%) for the Hotchkiss classification and 37.7% (95% CI 30.5%-44.9%) for the AO classification. The kappa statistic was 0.585 (0.541-0.661) for the Hotchkiss classification and 0.261 (0.240-0.350) for the AO classification. The mean percent agreement was 89.3% (86.6%-92.0%) for the consolidated Hotchkiss classification and 67.4% (54.6%-80.3%) for the consolidated AO classification. The kappa statistic was 0.760 (0.691-0.805) for the consolidated Hotchkiss classification and 0.455 (0.372-0.521) for the consolidated AO classification. CONCLUSION: The interobserver reliability for the Hotchkiss modification of the Mason classification was moderate, and that for the AO classification was fair according to the criteria of Landis and Koch. Collapsing the Hotchkiss classification improved the reliability to substantial, and collapsing the AO system improved reliability to the lower end of moderate. PMID- 19680512 TI - The who, what and when of surgery for the degenerative lumbar spine: a population based study of surgeon factors, surgical procedures, recent trends and reoperation rates. AB - BACKGROUND: Degenerative disease of the lumbar spine (DLS) is a common condition for which surgery can be beneficial in selected patients. With recent surgical trends toward more focused subspecialty training, it is unclear how characteristics of the surgical consultant may impact on treatment and reoperations. Our objective was to understand the relations between surgeon factors (who), surgical procedures (what) and recent trends (when) and their influence on reoperations for DLS surgery. METHODS: We performed a longitudinal population-based study using administrative databases including all patients aged 50 years and older who underwent surgery for DLS. We collected data on surgeon characteristics (specialty, volume), index procedures (decompressions, fusions) and reoperations. RESULTS: We identified 6128 patients who underwent surgery for DLS (4200 who had decompressions, 1928 who had fusions). We observed an increasing proportion of fusions over decompressions while the per capita surgeon supply declined. Orthopedic specialty and higher surgical volume were associated with a higher proportion of fusions (p < 0.001). The overall reoperation rate was 10.6%. Reoperations were more frequent in patients who had decompressions than those who had fusions at 2 years (5.4% v. 3.8%, odds ratio 1.4, p < 0.013), but not over the long-term. Long-term survival analysis demonstrated that a lower surgical volume was related to a higher reoperation rate (hazard ratio 1.28, p = 0.038). CONCLUSION: Lumbar spinal fusion rates for DLS have been increasing in Ontario. There is wide variation in surgical procedures between specialty and volume: namely high-volume and orthopedic surgeons have higer fusion rates than other surgeons. We observed better long-term survival among patients of high volume surgeons. Referring physicians should be aware that the choice of surgical consultant may influence patients' treatments and outcomes. With increasing rates of spinal surgery, the efficacy and cost benefit of current surgical options require ongoing study. PMID- 19680513 TI - Advanced laparoscopic training and outcomes in laparoscopic cholecystectomy. AB - BACKGROUND: Advanced laparoscopic training is becoming a valuable asset for surgeons as more procedures are carried out in a minimally invasive fashion. The purpose of our study was to determine whether laparoscopic fellowship training affects outcomes in patients undergoing laparoscopic cholecystectomy for acute cholecystitis. METHODS: We obtained data from a retrospective review of 110 patients with acute cholecystitis who underwent laparoscopic cholecystectomy on an urgent basis from March 2002 to June 2005. We compared the outcomes of 31 patients whose surgeries were performed by a surgeon with advanced laparoscopic training with those of 79 patients whose surgeries were performed by surgeons without such training. RESULTS: The 2 groups were similar in terms of demographics and time to surgery. Outcome measures included conversion rates, postoperative length of stay (LOS) and complications. There was a significant difference in conversion rates (3.2 % v. 16.5 %, p = 0.050) and postoperative LOS (1.77 v. 2.82 d, p < 0.006) between the 2 groups, but there was no difference in the rate of postoperative complications. There was no significant difference in conversion rates among the surgeons without advanced training (p = 0.64). CONCLUSION: Based on our results, laparoscopic cholecystectomy in acute cholecystitis is associated with improved outcomes when performed by a surgeon with fellowship training in laparoscopic surgery. PMID- 19680514 TI - Factors affecting perioperative blood loss and transfusion rates in primary total joint arthroplasty: a prospective analysis of 1642 patients. AB - BACKGROUND: In recent years, the use of low molecular weight heparins such as dalteparin has become attractive because of their ease of administration and superiority in preventing venous thromboembolism (VTE) compared with traditional agents. The primary purpose of our study was to evaluate the impact of dalteparin use on blood loss and transfusion rates in patients undergoing primary total joint arthroplasty. We also evaluated the effect of patient sex, releasing the tourniquet in knee arthroplasty and the turnover of house staff. METHODS: Using our hospital transfusion database, we prospectively studied the mean reduction in hemoglobin and transfusion rates of 1642 consecutive patients who underwent primary total hip arthroplasty (THA) or total knee arthroplasty (TKA) between January 2004 and December 2005. In 2004, warfarin was used exclusively for VTE prevention; however, in 2005, following the release of the 2004 American College of Chest Physicians' guidelines, our centre began using dalteparin for VTE prophylaxis. We analyzed the impact of dalteparin use and the effect of patient sex, tourniquet release in TKA and house staff turnover months on blood loss and transfusion rates. RESULTS: The use of dalteparin for postoperative VTE prevention in patients undergoing THA and TKA in 2005 was associated with a significantly greater mean reduction in hemoglobin compared with warfarin use in 2004 (p = 0.014 for patients undergoing THA, p < 0.001 for patients undergoing TKA). The use of dalteparin in 2005 was not associated with a significant increase in allogeneic blood transfusions compared with the use of warfarin in 2004, except in women (p < 0.001). Although we observed no significant differences in mean reduction in hemoglobin between men and women undergoing THA, women undergoing THA had significantly higher transfusion rates regardless of the method of VTE prophylaxis (p = 0.037 for warfarin, p < 0.001 for dalteparin). Intraoperative tourniquet release in patients undergoing TKA was associated with a significantly lower mean reduction in hemoglobin than release after wound closure (p = 0.005). Although house staff turnover months were associated with a significantly greater mean reduction in hemoglobin levels than non-turnover months (p = 0.039), these months were not associated with a significant increase in allogeneic blood transfusions (p = 0.59). CONCLUSION: Low molecular weight heparins such as dalteparin are the most common form of VTE prophylaxis in Canada. Our results suggest that dalteparin use, timing of tourniquet release and house staff turnover can all influence transfusion rates and/or blood loss in patients undergoing primary total joint arthroplasty. This study also emphasizes that women undergoing THA are at particularly high risk for blood transfusion. PMID- 19680515 TI - Mortality and complications following stabilization of femoral metastatic lesions: a population-based study of regional variation and outcome. AB - BACKGROUND: It is considered that patients at risk for spontaneous fracture due to metastatic lesions should undergo surgical stabilization before fracture occurs; however, prophylactic stabilization is associated with surgical morbidity and mortality. We sought to compare pathological fracture fixation versus prophylactic stabilization of diaphyseal femoral lesions for patients with femoral metastases and assess the rate of prophylactic surgery completed in all regions of Ontario. METHODS: Using population data sets, we identified all patients who had undergone femoral stabilization, either for pathological femoral fractures or for prophylactic fixation of femoral metastases before pathological fractures, between 1992 and 1997 in Ontario. We compared the rates of survival, serious medical and surgical complications and length of stay in hospital between the 2 groups. RESULTS: A total of 624 patients underwent surgical stabilization for femoral metastases. The most common sites of primary metastases were the lungs (26%), breasts (16%), kidneys (6%) and prostate (6%); 46% of patients had other or multiple primary metastases. Overall, 37% of lesions were fixed prophylactically, with wide variation by region (17.6%-72.2%). Patients who underwent prophylactic stabilization had better overall survival at all postoperative time points. This held true after adjusting for age, sex, comorbidities and type of cancer (p < 0.001). CONCLUSION: These data demonstrate a survival advantage with prophylactic fixation of metastatic femoral lesions combined with a relatively low perioperative risk excluding concomitant bilateral procedures. Ontario regional rates of prophylactic fixation vary enormously, with most patients not receiving prophylactic treatment. PMID- 19680516 TI - Evaluation of preoperative and perioperative operating room briefings at the Hospital for Sick Children. AB - BACKGROUND: Wrong-site, wrong-procedure and wrong-patient surgeries are catastrophic events for patients, medical caregivers and institutions. Operating room (OR) briefings are intended to reduce the risk of wrong-site surgeries and promote collaboration among OR personnel. The purpose of our study was to evaluate 2 OR briefing safety initiatives, "07:35 huddles" (preoperative OR briefing) and "surgical time-outs" (perioperative OR briefing), at the Hospital for Sick Children in Toronto, Ont. METHODS: First, we evaluated the completion and components of the 07:35 huddles and surgical time-outs briefings using direct observations. We then evaluated the attitudes of the OR staff regarding safety in the OR using the "Safety Attitudes Questionnaire, Operating Room version." Finally, we conducted personal interviews with OR personnel. RESULTS: Based on direct observations, 102 of 159 (64.1%) 07:35 huddles and 230 of 232 (99.1%) surgical time-outs briefings were completed. The perception of safety in the OR improved, but only among nurses. Regarding difficulty discussing errors in the OR, the nurses' mean scores improved from 3.5 (95% confidence interval [CI] 3.2 3.8) prebriefing to 2.8 (95% CI 2.5-3.2) postbriefing on a 5-point Likert scale (p < 0.05). Personal interviews confirmed that, mainly among the nursing staff, pre-and perioperative briefing tools increase the perception of communication within the OR, such that discussions regarding errors within the OR are more encouraged. CONCLUSION: Structured communication tools, such as 07:35 huddles and surgical time-outs briefings, especially for the nursing personnel, change the notion of individual advocacy to one of teamwork and being proactive about patient safety. PMID- 19680518 TI - The authors reply. PMID- 19680517 TI - Criteria for a diagnosis of abdominal compartment syndrome. PMID- 19680519 TI - The history of women in surgery. AB - The history of women in surgery in Western civilization dates to 3500 before common era (BCE) and Queen Shubad of Ur. Ancient history reveals an active role of women in surgery in Egypt, Italy and Greece as detailed in surgical texts of the time. During the middle ages, regulations forbade women from practising surgery unless they assumed their husbands' practices upon their deaths or unless they were deemed fit by a "competent" jury. King Henry VIII proclaimed that "No carpenter, smith, weaver or women shall practise surgery." The modern period of surgery opens with women impersonating men to practise medicine and surgery (Dr. Miranda Stewart). The first female physicians (Dr. Elizabeth Blackwell and Dr. Emily Jennings Stowe) and surgeons (Dr. Mary Edwards Walker and Dr. Jennie Smillie Robertson) in North America found it difficult to obtain residency education after completing medical school. Dr. Jessie Gray was Canada's "First Lady of Surgery" and the first woman to graduate from the Gallie program at the University of Toronto in the 1940s. Currently, the ratio of women in surgical training is far less than that of women in medical school. The reasons that women choose surgery include appropriate role models and intellectual/technical challenge. Lack of mentorship and lifestyle issues are the strongest deterrents. Consideration of a "controllable lifestyle" by surgical administrators will help with the recruitment of women into surgery. PMID- 19680521 TI - How to use an article about prognosis. PMID- 19680520 TI - Canadian consensus conference on the development of training and practice standards in advanced minimally invasive surgery: Edmonton, Alta., Jun. 1, 2007. AB - Despite the complexities of minimally invasive surgery (MIS), a Canadian approach to training surgeons in this field does not exist. Whereas a limited number of surgeons are fellowship-trained in the specialty, guidelines are still clearly needed to implement advanced MIS. Leaders in the field of gastrointestinal surgery and MIS attended a consensus conference where they proposed a comprehensive mentoring program that may evolve into a framework for a national mentoring and training system. Leadership and commitment from national experts to define the most appropriate template for introducing new surgical techniques into practice is required. This national framework should also provide flexibility for truly novel procedures such as natural orifice translumenal endoscopic surgery. PMID- 19680522 TI - Operating on a patient with hepatitis C. AB - The hepatitis C virus (HCV) infects more than 250 000 Canadians, many of whom are at increased risk of perioperative complications with surgery and all of whom represent an important risk for the entire surgical care team. The virus likely remains undiagnosed in one-third, while others have a limited understanding of the implications of the disease. All face an increased risk of liver failure and hepatocellular carcinoma. We review hepatitis C infection, including the challenges presented in operating on a patient with HCV infection and the inherent risks for the care team. PMID- 19680523 TI - Soft tissue case 61. AB - A 55-year-old man presented to the emergency department with a 12-hour history of severe crampy abdominal pain, nausea, vomiting and obstipation. The patient had a complex medical history, including coronary artery disease, lupus, hypothyroidism, epilepsy, pancreatitis and renal calculi. However, the patient had no history of a hernia or abdominal surgery. Physical examination revealed a temperature of 38.5 degrees C and a soft distended abdomen that was diffusely tender without signs of peritonitis. The rest of the physical examination was unremarkable. Routine laboratory investigations including a complete blood cell count, electrolytes, liver enzymes and amylase were normal, with the exception of a decreased hemoglobin level of 116 g/L. We ordered a plain abdominal radiograph (Fig. 1) and a contrast-enhanced computed tomography (CT) scan of his abdomen. What is your diagnosis? PMID- 19680524 TI - Online case notes. PMID- 19680526 TI - Effective testosterone suppression for prostate cancer: is there a best castration therapy? AB - Achieving and maintaining effective suppression of serum testosterone levels in men treated with androgen ablation is one of the essential strategies in the management of prostate cancer. Historically, a serum testosterone below 50 ng/dL was considered to be the castrate level. Current data suggest that the new target for either surgical or chemical castration is a serum testosterone level of lower than 20 ng/dL in an attempt to maximize therapeutic outcomes. Testosterone breakthrough and the acute-on-chronic effects of administration of a luteinizing hormone-releasing hormone analogue may cause testosterone levels to periodically rise, sometimes to noncastrate levels. The goal of androgen ablation is to identify those agents that will most consistently achieve and maintain the lowest testosterone levels possible. PMID- 19680525 TI - Will the evolution of overactive bladder delivery systems increase patient compliance? AB - The negative impact of overactive bladder (OAB) on daily quality of life drives the large market of pharmacotherapy targeted at symptoms of urinary frequency and urgency, with or without urinary urge incontinence. Currently, the primary pharmacologic treatment modality is aimed at modulation of the efferent muscarinic receptors (M2 and M3) predominant in detrusor smooth muscle and responsible for involuntary or unwanted bladder contractions. However, due to drug effects in the muscarinic receptors of the salivary glands and intestinal smooth muscle, as well as extensive first-pass metabolism in the liver and intestinal tract yielding parent drug metabolites, adverse side effects are common and can be quite bothersome. These issues, encountered with many of the oral antimuscarinic formulations, limit their tolerability and affect long-term patient compliance and satisfaction. Thus, the benefit of pharmacotherapy for OAB must be a balance between efficacy and tolerability, also known as therapeutic index. This article reviews the current pharmacologic delivery systems available for the treatment of OAB, patient compliance, and reasons for discontinuation of medication. PMID- 19680527 TI - Status of radical prostatectomy in 2009: is there medical evidence to justify the robotic approach? AB - This article presents the evolution of open radical retropubic prostatectomy (ORRP) into a minimally invasive procedure and reviews the literature to provide a legitimate comparison between ORRP and robotic-assisted laparoscopic radical retropubic prostatectomy (RALRP). The article is limited to manuscripts cited in the peer-reviewed literature, and an effort was made to identify those articles that fulfilled the highest level of medical evidence. In centers of excellence, ORRP is performed with no mortality, extraordinarily low technical and medical complications (1%), the rare need for blood transfusions, 1- to 2-day hospital stays, urinary catheters that are routinely removed in a week, the majority of men returning to work in 2 weeks, and up to 97% of men regaining urinary continence. Return of potency remains a challenge, especially for older men with marginal erections. RALRP is now the most common approach for the surgical removal of the malignant prostate. A critical review of the literature fails to support the marketing claims that RALRP is associated with shorter hospitalization, less pain, better cosmetics, shorter catheter time, lower transfusion rates, or improved continence and potency rates. The highest level of medical evidence suggests that RALRP may significantly compromise oncologic outcomes and that men undergoing this approach have higher regret rates than men undergoing ORRP. PMID- 19680529 TI - Best of the 2009 AUA Annual Meeting: Highlights from the 2009 Annual Meeting of the American Urological Association, April 25-30, 2009, Chicago, IL. PMID- 19680530 TI - Genitourinary sarcoidosis. AB - Genitourinary involvement of sarcoidosis can mimic many common urologic conditions. Although sarcoidosis is a benign inflammatory condition, it can present much like malignant or infectious conditions; thus, failed diagnosis can lead to unnecessary medications or surgical procedures. In addition, management choices for patients with scrotal findings have important implications for future fertility. Thus, this uncommon condition should be on the differential diagnosis for any urologic patient. The authors report on a patient with a scrotal mass as his presenting symptom of sarcoidosis and review the diagnosis, implications, and management of sarcoidosis involving all potential sites in the urinary tract. PMID- 19680528 TI - Spinal dysraphism: a neurosurgical review for the urologist. AB - Spinal neural tube defects are congenital malformations of the spine and spinal cord (eg, myelomeningocele) and are frequently seen in pediatric urology practice. These neurologic problems have many consequences in a child's life and affect different parts of the body, such as the brain, spinal cord, limbs, bladder, and bowels. Because of the complexity and neurologic aspects of spinal dysraphism, many related terms and aspects of the disease are unfamiliar to the urologist. This review addresses some of the most commonly used neurosurgical terms and concepts related to spinal dysraphism. PMID- 19680531 TI - Randomized trials of selenium, vitamin e, or vitamin C for prostate cancer prevention. PMID- 19680532 TI - Selective serotonin reuptake inhibitors cause erectile dysfunction: true or false? PMID- 19680533 TI - CHD3 proteins and polycomb group proteins antagonistically determine cell identity in Arabidopsis. AB - Dynamic regulation of chromatin structure is of fundamental importance for modulating genomic activities in higher eukaryotes. The opposing activities of Polycomb group (PcG) and trithorax group (trxG) proteins are part of a chromatin based cellular memory system ensuring the correct expression of specific transcriptional programs at defined developmental stages. The default silencing activity of PcG proteins is counteracted by trxG proteins that activate PcG target genes and prevent PcG mediated silencing activities. Therefore, the timely expression and regulation of PcG proteins and counteracting trxG proteins is likely to be of fundamental importance for establishing cell identity. Here, we report that the chromodomain/helicase/DNA-binding domain CHD3 proteins PICKLE (PKL) and PICKLE RELATED2 (PKR2) have trxG-like functions in plants and are required for the expression of many genes that are repressed by PcG proteins. The pkl mutant could partly suppress the leaf and flower phenotype of the PcG mutant curly leaf, supporting the idea that CHD3 proteins and PcG proteins antagonistically determine cell identity in plants. The direct targets of PKL in roots include the PcG genes SWINGER and EMBRYONIC FLOWER2 that encode subunits of Polycomb repressive complexes responsible for trimethylating histone H3 at lysine 27 (H3K27me3). Similar to mutants lacking PcG proteins, lack of PKL and PKR2 caused reduced H3K27me3 levels and, therefore, increased expression of a set of PcG protein target genes in roots. Thus, PKL and PKR2 are directly required for activation of PcG protein target genes and in roots are also indirectly required for repression of PcG protein target genes. Reduced PcG protein activity can lead to cell de-differentiation and callus-like tissue formation in pkl pkr2 mutants. Thus, in contrast to mammals, where PcG proteins are required to maintain pluripotency and to prevent cell differentiation, in plants PcG proteins are required to promote cell differentiation by suppressing embryonic development. PMID- 19680534 TI - Essential role of cyclophilin A for hepatitis C virus replication and virus production and possible link to polyprotein cleavage kinetics. AB - Viruses are obligate intracellular parasites and therefore their replication completely depends on host cell factors. In case of the hepatitis C virus (HCV), a positive-strand RNA virus that in the majority of infections establishes persistence, cyclophilins are considered to play an important role in RNA replication. Subsequent to the observation that cyclosporines, known to sequester cyclophilins by direct binding, profoundly block HCV replication in cultured human hepatoma cells, conflicting results were obtained as to the particular cyclophilin (Cyp) required for viral RNA replication and the underlying possible mode of action. By using a set of cell lines with stable knock-down of CypA or CypB, we demonstrate in the present work that replication of subgenomic HCV replicons of different genotypes is reduced by CypA depletion up to 1,000-fold whereas knock-down of CypB had no effect. Inhibition of replication was rescued by over-expression of wild type CypA, but not by a mutant lacking isomerase activity. Replication of JFH1-derived full length genomes was even more sensitive to CypA depletion as compared to subgenomic replicons and virus production was completely blocked. These results argue that CypA may target an additional viral factor outside of the minimal replicase contributing to RNA amplification and assembly, presumably nonstructural protein 2. By selecting for resistance against the cyclosporine analogue DEBIO-025 that targets CypA in a dose-dependent manner, we identified two mutations (V2440A and V2440L) close to the cleavage site between nonstructural protein 5A and the RNA-dependent RNA polymerase in nonstructural protein 5B that slow down cleavage kinetics at this site and reduce CypA dependence of viral replication. Further amino acid substitutions at the same cleavage site accelerating processing increase CypA dependence. Our results thus identify an unexpected correlation between HCV polyprotein processing and CypA dependence of HCV replication. PMID- 19680535 TI - Expression and processing of a small nucleolar RNA from the Epstein-Barr virus genome. AB - Small nucleolar RNAs (snoRNAs) are localized within the nucleolus, a sub-nuclear compartment, in which they guide ribosomal or spliceosomal RNA modifications, respectively. Up until now, snoRNAs have only been identified in eukaryal and archaeal genomes, but are notably absent in bacteria. By screening B lymphocytes for expression of non-coding RNAs (ncRNAs) induced by the Epstein-Barr virus (EBV), we here report, for the first time, the identification of a snoRNA gene within a viral genome, designated as v-snoRNA1. This genetic element displays all hallmark sequence motifs of a canonical C/D box snoRNA, namely C/C'- as well as D/D'-boxes. The nucleolar localization of v-snoRNA1 was verified by in situ hybridisation of EBV-infected cells. We also confirmed binding of the three canonical snoRNA proteins, fibrillarin, Nop56 and Nop58, to v-snoRNA1. The C-box motif of v-snoRNA1 was shown to be crucial for the stability of the viral snoRNA; its selective deletion in the viral genome led to a complete down-regulation of v snoRNA1 expression levels within EBV-infected B cells. We further provide evidence that v-snoRNA1 might serve as a miRNA-like precursor, which is processed into 24 nt sized RNA species, designated as v-snoRNA1(24pp). A potential target site of v-snoRNA1(24pp) was identified within the 3'-UTR of BALF5 mRNA which encodes the viral DNA polymerase. V-snoRNA1 was found to be expressed in all investigated EBV-positive cell lines, including lymphoblastoid cell lines (LCL). Interestingly, induction of the lytic cycle markedly up-regulated expression levels of v-snoRNA1 up to 30-fold. By a computational approach, we identified a v snoRNA1 homolog in the rhesus lymphocryptovirus genome. This evolutionary conservation suggests an important role of v-snoRNA1 during gamma-herpesvirus infection. PMID- 19680536 TI - Two HIV-1 variants resistant to small molecule CCR5 inhibitors differ in how they use CCR5 for entry. AB - HIV-1 variants resistant to small molecule CCR5 inhibitors recognize the inhibitor-CCR5 complex, while also interacting with free CCR5. The most common genetic route to resistance involves sequence changes in the gp120 V3 region, a pathway followed when the primary isolate CC1/85 was cultured with the AD101 inhibitor in vitro, creating the CC101.19 resistant variant. However, the D1/86.16 escape mutant contains no V3 changes but has three substitutions in the gp41 fusion peptide. By using CCR5 point-mutants and gp120-targeting agents, we have investigated how infectious clonal viruses derived from the parental and both resistant isolates interact with CCR5. We conclude that the V3 sequence changes in CC101.19 cl.7 create a virus with an increased dependency on interactions with the CCR5 N-terminus. Elements of the CCR5 binding site associated with the V3 region and the CD4-induced (CD4i) epitope cluster in the gp120 bridging sheet are more exposed on the native Env complex of CC101.19 cl.7, which is sensitive to neutralization via these epitopes. However, D1/86.16 cl.23 does not have an increased dependency on the CCR5 N-terminus, and its CCR5 binding site has not become more exposed. How this virus interacts with the inhibitor-CCR5 complex remains to be understood. PMID- 19680537 TI - Model-based deconvolution of cell cycle time-series data reveals gene expression details at high resolution. AB - In both prokaryotic and eukaryotic cells, gene expression is regulated across the cell cycle to ensure "just-in-time" assembly of select cellular structures and molecular machines. However, present in all time-series gene expression measurements is variability that arises from both systematic error in the cell synchrony process and variance in the timing of cell division at the level of the single cell. Thus, gene or protein expression data collected from a population of synchronized cells is an inaccurate measure of what occurs in the average single cell across a cell cycle. Here, we present a general computational method to extract "single-cell"-like information from population-level time-series expression data. This method removes the effects of 1) variance in growth rate and 2) variance in the physiological and developmental state of the cell. Moreover, this method represents an advance in the deconvolution of molecular expression data in its flexibility, minimal assumptions, and the use of a cross validation analysis to determine the appropriate level of regularization. Applying our deconvolution algorithm to cell cycle gene expression data from the dimorphic bacterium Caulobacter crescentus, we recovered critical features of cell cycle regulation in essential genes, including ctrA and ftsZ, that were obscured in population-based measurements. In doing so, we highlight the problem with using population data alone to decipher cellular regulatory mechanisms and demonstrate how our deconvolution algorithm can be applied to produce a more realistic picture of temporal regulation in a cell. PMID- 19680538 TI - Statistical estimation of correlated genome associations to a quantitative trait network. AB - Many complex disease syndromes, such as asthma, consist of a large number of highly related, rather than independent, clinical or molecular phenotypes. This raises a new technical challenge in identifying genetic variations associated simultaneously with correlated traits. In this study, we propose a new statistical framework called graph-guided fused lasso (GFlasso) to directly and effectively incorporate the correlation structure of multiple quantitative traits such as clinical metrics and gene expressions in association analysis. Our approach represents correlation information explicitly among the quantitative traits as a quantitative trait network (QTN) and then leverages this network to encode structured regularization functions in a multivariate regression model over the genotypes and traits. The result is that the genetic markers that jointly influence subgroups of highly correlated traits can be detected jointly with high sensitivity and specificity. While most of the traditional methods examined each phenotype independently and combined the results afterwards, our approach analyzes all of the traits jointly in a single statistical framework. This allows our method to borrow information across correlated phenotypes to discover the genetic markers that perturb a subset of the correlated traits synergistically. Using simulated datasets based on the HapMap consortium and an asthma dataset, we compared the performance of our method with other methods based on single-marker analysis and regression-based methods that do not use any of the relational information in the traits. We found that our method showed an increased power in detecting causal variants affecting correlated traits. Our results showed that, when correlation patterns among traits in a QTN are considered explicitly and directly during a structured multivariate genome association analysis using our proposed methods, the power of detecting true causal SNPs with possibly pleiotropic effects increased significantly without compromising performance on non-pleiotropic SNPs. PMID- 19680539 TI - RNA gain-of-function in spinocerebellar ataxia type 8. AB - Microsatellite expansions cause a number of dominantly-inherited neurological diseases. Expansions in coding-regions cause protein gain-of-function effects, while non-coding expansions produce toxic RNAs that alter RNA splicing activities of MBNL and CELF proteins. Bi-directional expression of the spinocerebellar ataxia type 8 (SCA8) CTG CAG expansion produces CUG expansion RNAs (CUG(exp)) from the ATXN8OS gene and a nearly pure polyglutamine expansion protein encoded by ATXN8 CAG(exp) transcripts expressed in the opposite direction. Here, we present three lines of evidence that RNA gain-of-function plays a significant role in SCA8: 1) CUG(exp) transcripts accumulate as ribonuclear inclusions that co-localize with MBNL1 in selected neurons in the brain; 2) loss of Mbnl1 enhances motor deficits in SCA8 mice; 3) SCA8 CUG(exp) transcripts trigger splicing changes and increased expression of the CUGBP1-MBNL1 regulated CNS target, GABA-A transporter 4 (GAT4/Gabt4). In vivo optical imaging studies in SCA8 mice confirm that Gabt4 upregulation is associated with the predicted loss of GABAergic inhibition within the granular cell layer. These data demonstrate that CUG(exp) transcripts dysregulate MBNL/CELF regulated pathways in the brain and provide mechanistic insight into the CNS effects of other CUG(exp) disorders. Moreover, our demonstration that relatively short CUG(exp) transcripts cause RNA gain-of-function effects and the growing number of antisense transcripts recently reported in mammalian genomes suggest unrecognized toxic RNAs contribute to the pathophysiology of polyglutamine CAG CTG disorders. PMID- 19680540 TI - A mouse model for the metabolic effects of the human fat mass and obesity associated FTO gene. AB - Human FTO gene variants are associated with body mass index and type 2 diabetes. Because the obesity-associated SNPs are intronic, it is unclear whether changes in FTO expression or splicing are the cause of obesity or if regulatory elements within intron 1 influence upstream or downstream genes. We tested the idea that FTO itself is involved in obesity. We show that a dominant point mutation in the mouse Fto gene results in reduced fat mass, increased energy expenditure, and unchanged physical activity. Exposure to a high-fat diet enhances lean mass and lowers fat mass relative to control mice. Biochemical studies suggest the mutation occurs in a structurally novel domain and modifies FTO function, possibly by altering its dimerisation state. Gene expression profiling revealed increased expression of some fat and carbohydrate metabolism genes and an improved inflammatory profile in white adipose tissue of mutant mice. These data provide direct functional evidence that FTO is a causal gene underlying obesity. Compared to the reported mouse FTO knockout, our model more accurately reflects the effect of human FTO variants; we observe a heterozygous as well as homozygous phenotype, a smaller difference in weight and adiposity, and our mice do not show perinatal lethality or an age-related reduction in size and length. Our model suggests that a search for human coding mutations in FTO may be informative and that inhibition of FTO activity is a possible target for the treatment of morbid obesity. PMID- 19680541 TI - CLRN1 is nonessential in the mouse retina but is required for cochlear hair cell development. AB - Mutations in the CLRN1 gene cause Usher syndrome type 3 (USH3), a human disease characterized by progressive blindness and deafness. Clarin 1, the protein product of CLRN1, is a four-transmembrane protein predicted to be associated with ribbon synapses of photoreceptors and cochlear hair cells, and recently demonstrated to be associated with the cytoskeleton. To study Clrn1, we created a Clrn1 knockout (KO) mouse and characterized the histological and functional consequences of Clrn1 deletion in the retina and cochlea. Clrn1 KO mice do not develop a retinal degeneration phenotype, but exhibit progressive loss of sensory hair cells in the cochlea and deterioration of the organ of Corti by 4 months. Hair cell stereocilia in KO animals were longer and disorganized by 4 months, and some Clrn1 KO mice exhibited circling behavior by 5-6 months of age. Clrn1 mRNA expression was localized in the retina using in situ hybridization (ISH), laser capture microdissection (LCM), and RT-PCR. Retinal Clrn1 transcripts were found throughout development and adulthood by RT-PCR, although expression peaked at P7 and declined to undetectable levels in adult retina by ISH. LCM localized Clrn1 transcripts to the retinas inner nuclear layer, and WT levels of retinal Clrn1 expression were observed in photoreceptor-less retinas. Examination of Clrn1 KO mice suggests that CLRN1 is unnecessary in the murine retina but essential for normal cochlear development and function. This may reflect a redundancy in the mouse retina not present in human retina. In contrast to mouse KO models of USH1 and USH2, our data indicate that Clrn1 expression in the retina is restricted to the Muller glia. This is a novel finding, as most retinal degeneration associated proteins are expressed in photoreceptors, not in glia. If CLRN1 expression in humans is comparable to the expression pattern observed in mice, this is the first report of an inner retinal protein that, when mutated, causes retinal degeneration. PMID- 19680542 TI - Tissue effect on genetic control of transcript isoform variation. AB - Current genome-wide association studies (GWAS) are moving towards the use of large cohorts of primary cell lines to study a disease of interest and to assign biological relevance to the genetic signals identified. Here, we use a panel of human osteoblasts (HObs) to carry out a transcriptomic survey, similar to recent studies in lymphoblastoid cell lines (LCLs). The distinct nature of HObs and LCLs is reflected by the preferential grouping of cell type-specific genes within biologically and functionally relevant pathways unique to each tissue type. We performed cis-association analysis with SNP genotypes to identify genetic variations of transcript isoforms, and our analysis indicates that differential expression of transcript isoforms in HObs is also partly controlled by cis regulatory genetic variants. These isoforms are regulated by genetic variants in both a tissue-specific and tissue-independent fashion, and these associations have been confirmed by RT-PCR validation. Our study suggests that multiple transcript isoforms are often present in both tissues and that genetic control may affect the relative expression of one isoform to another, rather than having an all-or-none effect. Examination of the top SNPs from a GWAS of bone mineral density show overlap with probeset associations observed in this study. The top hit corresponding to the FAM118A gene was tested for association studies in two additional clinical studies, revealing a novel transcript isoform variant. Our approach to examining transcriptome variation in multiple tissue types is useful for detecting the proportion of genetic variation common to different cell types and for the identification of cell-specific isoform variants that may be functionally relevant, an important follow-up step for GWAS. PMID- 19680543 TI - A computational screen for regulators of oxidative phosphorylation implicates SLIRP in mitochondrial RNA homeostasis. AB - The human oxidative phosphorylation (OxPhos) system consists of approximately 90 proteins encoded by nuclear and mitochondrial genomes and serves as the primary cellular pathway for ATP biosynthesis. While the core protein machinery for OxPhos is well characterized, many of its assembly, maturation, and regulatory factors remain unknown. We exploited the tight transcriptional control of the genes encoding the core OxPhos machinery to identify novel regulators. We developed a computational procedure, which we call expression screening, which integrates information from thousands of microarray data sets in a principled manner to identify genes that are consistently co-expressed with a target pathway across biological contexts. We applied expression screening to predict dozens of novel regulators of OxPhos. For two candidate genes, CHCHD2 and SLIRP, we show that silencing with RNAi results in destabilization of OxPhos complexes and a marked loss of OxPhos enzymatic activity. Moreover, we show that SLIRP plays an essential role in maintaining mitochondrial-localized mRNA transcripts that encode OxPhos protein subunits. Our findings provide a catalogue of potential novel OxPhos regulators that advance our understanding of the coordination between nuclear and mitochondrial genomes for the regulation of cellular energy metabolism. PMID- 19680544 TI - Inositol 1,4,5- trisphosphate receptor function in Drosophila insulin producing cells. AB - The Inositol 1,4,5- trisphosphate receptor (InsP(3)R) is an intracellular ligand gated channel that releases calcium from intracellular stores in response to extracellular signals. To identify and understand physiological processes and behavior that depends on the InsP(3) signaling pathway at a systemic level, we are studying Drosophila mutants for the InsP(3)R (itpr) gene. Here, we show that growth defects precede larval lethality and both are a consequence of the inability to feed normally. Moreover, restoring InsP(3)R function in insulin producing cells (IPCs) in the larval brain rescues the feeding deficit, growth and lethality in the itpr mutants to a significant extent. We have previously demonstrated a critical requirement for InsP(3)R activity in neuronal cells, specifically in aminergic interneurons, for larval viability. Processes from the IPCs and aminergic domain are closely apposed in the third instar larval brain with no visible cellular overlap. Ubiquitous depletion of itpr by dsRNA results in feeding deficits leading to larval lethality similar to the itpr mutant phenotype. However, when itpr is depleted specifically in IPCs or aminergic neurons, the larvae are viable. These data support a model where InsP(3)R activity in non-overlapping neuronal domains independently rescues larval itpr phenotypes by non-cell autonomous mechanisms. PMID- 19680545 TI - Continuous multi-parameter heart rate variability analysis heralds onset of sepsis in adults. AB - BACKGROUND: Early diagnosis of sepsis enables timely resuscitation and antibiotics and prevents subsequent morbidity and mortality. Clinical approaches relying on point-in-time analysis of vital signs or lab values are often insensitive, non-specific and late diagnostic markers of sepsis. Exploring otherwise hidden information within intervals-in-time, heart rate variability (HRV) has been documented to be both altered in the presence of sepsis, and correlated with its severity. We hypothesized that by continuously tracking individual patient HRV over time in patients as they develop sepsis, we would demonstrate reduced HRV in association with the onset of sepsis. METHODOLOGY/PRINCIPAL FINDINGS: We monitored heart rate continuously in adult bone marrow transplant (BMT) patients (n = 21) beginning a day before their BMT and continuing until recovery or withdrawal (12+/-4 days). We characterized HRV continuously over time with a panel of time, frequency, complexity, and scale invariant domain techniques. We defined baseline HRV as mean variability for the first 24 h of monitoring and studied individual and population average percentage change (from baseline) over time in diverse HRV metrics, in comparison with the time of clinical diagnosis and treatment of sepsis (defined as systemic inflammatory response syndrome along with clinically suspected infection requiring treatment). Of the 21 patients enrolled, 4 patients withdrew, leaving 17 patients who completed the study. Fourteen patients developed sepsis requiring antibiotic therapy, whereas 3 did not. On average, for 12 out of 14 infected patients, a significant (25%) reduction prior to the clinical diagnosis and treatment of sepsis was observed in standard deviation, root mean square successive difference, sample and multiscale entropy, fast Fourier transform, detrended fluctuation analysis, and wavelet variability metrics. For infected patients (n = 14), wavelet HRV demonstrated a 25% drop from baseline 35 h prior to sepsis on average. For 3 out of 3 non-infected patients, all measures, except root mean square successive difference and entropy, showed no significant reduction. Significant correlation was present amongst these HRV metrics for the entire population. CONCLUSIONS/SIGNIFICANCE: Continuous HRV monitoring is feasible in ambulatory patients, demonstrates significant HRV alteration in individual patients in association with, and prior to clinical diagnosis and treatment of sepsis, and merits further investigation as a means of providing early warning of sepsis. PMID- 19680546 TI - Epigenetic engineering of ribosomal RNA genes enhances protein production. AB - Selection of mammalian high-producer cell lines remains a major challenge for the biopharmaceutical manufacturing industry. Ribosomal RNA (rRNA) genes encode the major component of the ribosome but many rRNA gene copies are not transcribed due to epigenetic silencing by the nucleolar remodelling complex (NoRC) [6], which may limit the cell's full production capacity. Here we show that the knockdown of TIP5, a subunit of NoRC, decreases the number of silent rRNA genes, upregulates rRNA transcription, enhances ribosome synthesis and increases production of recombinant proteins. However, general enhancement of rRNA transcription rate did not stimulate protein synthesis. Our data demonstrates that the number of transcriptionally competent rRNA genes limits efficient ribosome synthesis. Epigenetic engineering of ribosomal RNA genes offers new possibilities for improving biopharmaceutical manufacturing and provides novel insights into the complex regulatory network which governs the translation machinery in normal cellular processes as well as in pathological conditions like cancer. PMID- 19680547 TI - Deregulation of hepatic insulin sensitivity induced by central lipid infusion in rats is mediated by nitric oxide. AB - BACKGROUND: Deregulation of hypothalamic fatty acid sensing lead to hepatic insulin-resistance which may partly contribute to further impairment of glucose homeostasis. METHODOLOGY: We investigated here whether hypothalamic nitric oxide (NO) could mediate deleterious peripheral effect of central lipid overload. Thus we infused rats for 24 hours into carotid artery towards brain, either with heparinized triglyceride emulsion (Intralipid, IL) or heparinized saline (control rats). PRINCIPAL FINDINGS: Lipids infusion led to hepatic insulin-resistance partly related to a decreased parasympathetic activity in the liver assessed by an increased acetylcholinesterase activity. Hypothalamic nitric oxide synthases (NOS) activities were significantly increased in IL rats, as the catalytically active neuronal NOS (nNOS) dimers compared to controls. This was related to a decrease in expression of protein inhibitor of nNOS (PIN). Effect of IL infusion on deregulated hepatic insulin-sensitivity was reversed by carotid injection of non selective NOS inhibitor NG-monomethyl-L-arginine (L-NMMA) and also by a selective inhibitor of the nNOS isoform, 7-Nitro-Indazole (7-Ni). In addition, NO donor injection (L-arginine and SNP) within carotid in control rats mimicked lipid effects onto impaired hepatic insulin sensitivity. In parallel we showed that cultured VMH neurons produce NO in response to fatty acid (oleic acid). CONCLUSIONS/SIGNIFICANCE: We conclude that cerebral fatty acid overload induces an enhancement of nNOS activity within hypothalamus which is, at least in part, responsible fatty acid increased hepatic glucose production. PMID- 19680549 TI - Timed somatic deletion of p53 in mice reveals age-associated differences in tumor progression. AB - Inactivating mutations in the p53 tumor suppressor gene occur often in the progression of human cancers. p53 inhibits the outgrowth of nascent cancer cells through anti-proliferative actions (including induction of apoptosis or senescence). To test p53 tumor suppressor functions in a novel experimental context, we somatically deleted both p53 alleles in multiple tissues of mice at various ages. Mice homozygously deleted for p53 at 3 months of age showed a longer tumor latency compared to mice deleted for p53 at 6 and 12 months of age. These results are consistent with a model in which tissues accumulate oncogenically activated cells with age and these are held in check by wildtype p53. We also deleted p53 before, concurrent with, and after treatment of mice with ionizing radiation (IR). The absence or presence of p53 during IR treatment had no effect on radiation-induced lymphoma latency, confirming that the immediate p53 damage response was irrelevant for cancer prevention. Even the presence of wildtype p53 for up to four weeks post-IR provided no protection against early lymphoma incidence, indicating that long term maintenance of functional p53 is critical for preventing the emergence of a cancer. These experiments indicate that sustained p53 anti-oncogenic function acts as a final or near final line of defense preventing progression of oncogenically activated cells to malignant tumors. PMID- 19680548 TI - Phosphodiesterase inhibition increases CREB phosphorylation and restores orientation selectivity in a model of fetal alcohol spectrum disorders. AB - BACKGROUND: Fetal alcohol spectrum disorders (FASD) are the leading cause of mental retardation in the western world and children with FASD present altered somatosensory, auditory and visual processing. There is growing evidence that some of these sensory processing problems may be related to altered cortical maps caused by impaired developmental neuronal plasticity. METHODOLOGY/PRINCIPAL FINDINGS: Here we show that the primary visual cortex of ferrets exposed to alcohol during the third trimester equivalent of human gestation have decreased CREB phosphorylation and poor orientation selectivity revealed by western blotting, optical imaging of intrinsic signals and single-unit extracellular recording techniques. Treating animals several days after the period of alcohol exposure with a phosphodiesterase type 1 inhibitor (Vinpocetine) increased CREB phosphorylation and restored orientation selectivity columns and neuronal orientation tuning. CONCLUSIONS/SIGNIFICANCE: These findings suggest that CREB function is important for the maturation of orientation selectivity and that plasticity enhancement by vinpocetine may play a role in the treatment of sensory problems in FASD. PMID- 19680551 TI - Immature and maturation-resistant human dendritic cells generated from bone marrow require two stimulations to induce T cell anergy in vitro. AB - Immature dendritic cells (DC) represent potential clinical tools for tolerogenic cellular immunotherapy in both transplantation and autoimmunity. A major drawback in vivo is their potential to mature during infections or inflammation, which would convert their tolerogenicity into immunogenicity. The generation of immature DC from human bone marrow (BM) by low doses of GM-CSF (lowGM) in the absence of IL-4 under GMP conditions create DC resistant to maturation, detected by surface marker expression and primary stimulation by allogeneic T cells. This resistence could not be observed for BM-derived DC generated with high doses of GM-CSF plus IL-4 (highGM/4), although both DC types induced primary allogeneic T cell anergy in vitro. The estabishment of the anergic state requires two subsequent stimulations by immature DC. Anergy induction was more profound with lowGM-DC due to their maturation resistance. Together, we show the generation of immature, maturation-resistant lowGM-DC for potential clinical use in transplant rejection and propose a two-step-model of T cell anergy induction by immature DC. PMID- 19680550 TI - Function of SSA subfamily of Hsp70 within and across species varies widely in complementing Saccharomyces cerevisiae cell growth and prion propagation. AB - BACKGROUND: The cytosol of most eukaryotic cells contains multiple highly conserved Hsp70 orthologs that differ mainly by their spatio-temporal expression patterns. Hsp70s play essential roles in protein folding, transport or degradation, and are major players of cellular quality control processes. However, while several reports suggest that specialized functions of Hsp70 orthologs were selected through evolution, few studies addressed systematically this issue. METHODOLOGY/PRINCIPAL FINDINGS: We compared the ability of Ssa1p Ssa4p from Saccharomyces cerevisiae and Ssa5p-Ssa8p from the evolutionary distant yeast Yarrowia lipolytica to perform Hsp70-dependent tasks when expressed as the sole Hsp70 for S. cerevisiae in vivo. We show that Hsp70 isoforms (i) supported yeast viability yet with markedly different growth rates, (ii) influenced the propagation and stability of the [PSI(+)] and [URE3] prions, but iii) did not significantly affect the proteasomal degradation rate of CFTR. Additionally, we show that individual Hsp70 orthologs did not induce the formation of different prion strains, but rather influenced the aggregation properties of Sup35 in vivo. Finally, we show that [URE3] curing by the overexpression of Ydj1p is Hsp70 isoform dependent. CONCLUSION/SIGNIFICANCE: Despite very high homology and overlapping functions, the different Hsp70 orthologs have evolved to possess distinct activities that are required to cope with different types of substrates or stress situations. Yeast prions provide a very sensitive model to uncover this functional specialization and to explore the intricate network of chaperone/co chaperone/substrates interactions. PMID- 19680552 TI - CK2 is the regulator of SIRT1 substrate-binding affinity, deacetylase activity and cellular response to DNA-damage. AB - SIRT1, an NAD(+) (nicotinamide adenine dinucleotide)-dependent deacetylase, protects cells from stress-induced apoptosis, and its orthologues delay aging in lower eukaryotes. SIRT1 increases survival in response to stress such as DNA damage by deacetylating a number of substrates including pro-apoptotic protein p53. The molecular mechanism by which DNA-damage activates SIRT1 is not known. By screening a kinase inhibitor library, we identified CK2 as a SIRT1 kinase. CK2 is a pleiotropic kinase with more than 300 substrates and well-known anti-apoptotic and pro-growth activities. We find that CK2 is recruited to SIRT1 after ionizing radiation (IR) and phosphorylates conserved residues Ser 154, 649, 651 and 683 in the N- and C-terminal domains of mouse SIRT1. Phosphorylation of SIRT1 increases its deacetylation rate but not if the four Ser residues are mutated. In addition, phosphorylation of SIRT1 increases its substrate-binding affinity. CK2-mediated phosphorylation increases the ability of SIRT1 to deacetylate p53 and protect cells from apoptosis after DNA damage. Based on these findings, we propose that CK2 protects against IR-induced apoptosis partly by phosphorylating and activating SIRT1. Thus, this work suggests that SIRT1 is a component of the expansive anti-apoptotic network controlled by CK2. Since expression of both CK2 and SIRT1 is upregulated with tumorigenesis and downregulated with senescence, the CK2-SIRT1 link sheds new light on how CK2 may regulate cancer development and aging. PMID- 19680553 TI - Endogenous human brain dynamics recover slowly following cognitive effort. AB - BACKGROUND: In functional magnetic resonance imaging, the brain's response to experimental manipulation is almost always assumed to be independent of endogenous oscillations. To test this, we addressed the possible interaction between cognitive task performance and endogenous fMRI oscillations in an experiment designed to answer two questions: 1) Does performance of a cognitively effortful task significantly change fractal scaling properties of fMRI time series compared to their values before task performance? 2) If so, can we relate the extent of task-related perturbation to the difficulty of the task? METHODOLOGY/PRINCIPAL FINDINGS: Using a novel continuous acquisition "rest-task rest" design, we found that endogenous dynamics tended to recover their pre-task parameter values relatively slowly, over the course of several minutes, following completion of one of two versions of the n-back working memory task and that the rate of recovery was slower following completion of the more demanding (n = 2) version of the task. CONCLUSION/SIGNIFICANCE: This result supports the model that endogenous low frequency oscillatory dynamics are relevant to the brain's response to exogenous stimulation. Moreover, it suggests that large-scale neurocognitive systems measured using fMRI, like the heart and other physiological systems subjected to external demands for enhanced performance, can take a considerable period of time to return to a stable baseline state. PMID- 19680554 TI - Sterol Biosynthesis Pathway as Target for Anti-trypanosomatid Drugs. AB - Sterols are constituents of the cellular membranes that are essential for their normal structure and function. In mammalian cells, cholesterol is the main sterol found in the various membranes. However, other sterols predominate in eukaryotic microorganisms such as fungi and protozoa. It is now well established that an important metabolic pathway in fungi and in members of the Trypanosomatidae family is one that produces a special class of sterols, including ergosterol, and other 24-methyl sterols, which are required for parasitic growth and viability, but are absent from mammalian host cells. Currently, there are several drugs that interfere with sterol biosynthesis (SB) that are in use to treat diseases such as high cholesterol in humans and fungal infections. In this review, we analyze the effects of drugs such as (a) statins, which act on the mevalonate pathway by inhibiting HMG-CoA reductase, (b) bisphosphonates, which interfere with the isoprenoid pathway in the step catalyzed by farnesyl diphosphate synthase, (c) zaragozic acids and quinuclidines, inhibitors of squalene synthase (SQS), which catalyzes the first committed step in sterol biosynthesis, (d) allylamines, inhibitors of squalene epoxidase, (e) azoles, which inhibit C14alpha-demethylase, and (f) azasterols, which inhibit Delta(24(25))-sterol methyltransferase (SMT). Inhibition of this last step appears to have high selectivity for fungi and trypanosomatids, since this enzyme is not found in mammalian cells. We review here the IC50 values of these various inhibitors, their effects on the growth of trypanosomatids (both in axenic cultures and in cell cultures), and their effects on protozoan structural organization (as evaluted by light and electron microscopy) and lipid composition. The results show that the mitochondrial membrane as well as the membrane lining the protozoan cell body and flagellum are the main targets. Probably as a consequence of these primary effects, other important changes take place in the organization of the kinetoplast DNA network and on the protozoan cell cycle. In addition, apoptosis-like and autophagic processes induced by several of the inhibitors tested led to parasite death. PMID- 19680556 TI - Genetic variation in healthy oldest-old. AB - Individuals who live to 85 and beyond without developing major age-related diseases may achieve this, in part, by lacking disease susceptibility factors, or by possessing resistance factors that enhance their ability to avoid disease and prolong lifespan. Healthy aging is a complex phenotype likely to be affected by both genetic and environmental factors. We sequenced 24 candidate healthy aging genes in DNA samples from 47 healthy individuals aged eighty-five years or older (the 'oldest-old'), to characterize genetic variation that is present in this exceptional group. These healthy seniors were never diagnosed with cancer, cardiovascular disease, pulmonary disease, diabetes, or Alzheimer disease. We re sequenced all exons, intron-exon boundaries and selected conserved non-coding sequences of candidate genes involved in aging-related processes, including dietary restriction (PPARG, PPARGC1A, SIRT1, SIRT3, UCP2, UCP3), metabolism (IGF1R, APOB, SCD), autophagy (BECN1, FRAP1), stem cell activation (NOTCH1, DLL1), tumor suppression (TP53, CDKN2A, ING1), DNA methylation (TRDMT1, DNMT3A, DNMT3B) Progeria syndromes (LMNA, ZMPSTE24, KL) and stress response (CRYAB, HSPB2). We detected 935 variants, including 848 single nucleotide polymorphisms (SNPs) and 87 insertion or deletions; 41% (385) were not recorded in dbSNP. This study is the first to present a comprehensive analysis of genetic variation in aging-related candidate genes in healthy oldest-old. These variants and especially our novel polymorphisms are valuable resources to test for genetic association in models of disease susceptibility or resistance. In addition, we propose an innovative tagSNP selection strategy that combines variants identified through gene re-sequencing- and HapMap-derived SNPs. PMID- 19680555 TI - Diversity and strain specificity of plant cell wall degrading enzymes revealed by the draft genome of Ruminococcus flavefaciens FD-1. AB - BACKGROUND: Ruminococcus flavefaciens is a predominant cellulolytic rumen bacterium, which forms a multi-enzyme cellulosome complex that could play an integral role in the ability of this bacterium to degrade plant cell wall polysaccharides. Identifying the major enzyme types involved in plant cell wall degradation is essential for gaining a better understanding of the cellulolytic capabilities of this organism as well as highlighting potential enzymes for application in improvement of livestock nutrition and for conversion of cellulosic biomass to liquid fuels. METHODOLOGY/PRINCIPAL FINDINGS: The R. flavefaciens FD-1 genome was sequenced to 29x-coverage, based on pulsed-field gel electrophoresis estimates (4.4 Mb), and assembled into 119 contigs providing 4,576,399 bp of unique sequence. As much as 87.1% of the genome encodes ORFs, tRNA, rRNAs, or repeats. The GC content was calculated at 45%. A total of 4,339 ORFs was detected with an average gene length of 918 bp. The cellulosome model for R. flavefaciens was further refined by sequence analysis, with at least 225 dockerin-containing ORFs, including previously characterized cohesin-containing scaffoldin molecules. These dockerin-containing ORFs encode a variety of catalytic modules including glycoside hydrolases (GHs), polysaccharide lyases, and carbohydrate esterases. Additionally, 56 ORFs encode proteins that contain carbohydrate-binding modules (CBMs). Functional microarray analysis of the genome revealed that 56 of the cellulosome-associated ORFs were up-regulated, 14 were down-regulated, 135 were unaffected, when R. flavefaciens FD-1 was grown on cellulose versus cellobiose. Three multi-modular xylanases (ORF01222, ORF03896, and ORF01315) exhibited the highest levels of up-regulation. CONCLUSIONS/SIGNIFICANCE: The genomic evidence indicates that R. flavefaciens FD 1 has the largest known number of fiber-degrading enzymes likely to be arranged in a cellulosome architecture. Functional analysis of the genome has revealed that the growth substrate drives expression of enzymes predicted to be involved in carbohydrate metabolism as well as expression and assembly of key cellulosomal enzyme components. PMID- 19680557 TI - Genome-wide mRNA expression analysis of hepatic adaptation to high-fat diets reveals switch from an inflammatory to steatotic transcriptional program. AB - BACKGROUND: Excessive exposure to dietary fats is an important factor in the initiation of obesity and metabolic syndrome associated pathologies. The cellular processes associated with the onset and progression of diet-induced metabolic syndrome are insufficiently understood. PRINCIPAL FINDINGS: To identify the mechanisms underlying the pathological changes associated with short and long term exposure to excess dietary fat, hepatic gene expression of ApoE3Leiden mice fed chow and two types of high-fat (HF) diets was monitored using microarrays during a 16-week period. A functional characterization of 1663 HF-responsive genes reveals perturbations in lipid, cholesterol and oxidative metabolism, immune and inflammatory responses and stress-related pathways. The major changes in gene expression take place during the early (day 3) and late (week 12) phases of HF feeding. This is also associated with characteristic opposite regulation of many HF-affected pathways between these two phases. The most prominent switch occurs in the expression of inflammatory/immune pathways (early activation, late repression) and lipogenic/adipogenic pathways (early repression, late activation). Transcriptional network analysis identifies NF-kappaB, NEMO, Akt, PPARgamma and SREBP1 as the key controllers of these processes and suggests that direct regulatory interactions between these factors may govern the transition from early (stressed, inflammatory) to late (pathological, steatotic) hepatic adaptation to HF feeding. This transition observed by hepatic gene expression analysis is confirmed by expression of inflammatory proteins in plasma and the late increase in hepatic triglyceride content. In addition, the genes most predictive of fat accumulation in liver during 16-week high-fat feeding period are uncovered by regression analysis of hepatic gene expression and triglyceride levels. CONCLUSIONS: The transition from an inflammatory to a steatotic transcriptional program, possibly driven by the reciprocal activation of NF kappaB and PPARgamma regulators, emerges as the principal signature of the hepatic adaptation to excess dietary fat. These findings may be of essential interest for devising new strategies aiming to prevent the progression of high fat diet induced pathologies. PMID- 19680559 TI - Hypothalamic neuroendocrine functions in rats with dihydrotestosterone-induced polycystic ovary syndrome: effects of low-frequency electro-acupuncture. AB - Adult female rats continuously exposed to androgens from prepuberty have reproductive and metabolic features of polycystic ovary syndrome (PCOS). We investigated whether such exposure adversely affects estrous cyclicity and the expression and distribution of gonadotropin-releasing hormone (GnRH), GnRH receptors, and corticotrophin-releasing hormone (CRH) in the hypothalamus and whether the effects are mediated by the androgen receptor (AR). We also assessed the effect of low-frequency electro-acupuncture (EA) on those variables. At 21 days of age, rats were randomly divided into three groups (control, PCOS, and PCOS EA; n = 12/group) and implanted subcutaneously with 90-day continuous release pellets containing vehicle or 5alpha-dihydrostestosterone (DHT). From age 70 days, PCOS EA rats received 2-Hz EA (evoking muscle twitches) five times/week for 4-5 weeks. Hypothalamic protein expression was measured by immunohistochemistry and western blot. DHT-treated rats were acyclic, but controls had regular estrous cycles. In PCOS rats, hypothalamic medial preoptic AR protein expression and the number of AR- and GnRH-immunoreactive cells were increased, but CRH was not affected; however, GnRH receptor expression was decreased in both the pituitary and hypothalamus. Low-frequency EA restored estrous cyclicity within 1 week and reduced the elevated hypothalamic GnRH and AR expression levels. EA did not affect GnRH receptor or CRH expression. Interestingly, nuclear AR co-localized with GnRH in the hypothalamus. Thus, rats with DHT-induced PCOS have disrupted estrous cyclicity and an increased number of hypothalamic cells expressing GnRH, most likely mediated by AR activation. Repeated low-frequency EA normalized estrous cyclicity and restored GnRH and AR protein expression. These results may help explain the beneficial neuroendocrine effects of low-frequency EA in women with PCOS. PMID- 19680560 TI - Chances and limitations of wild bird monitoring for the avian influenza virus H5N1--detection of pathogens highly mobile in time and space. AB - Highly pathogenic influenza virus (HPAIV) H5N1 proved to be remarkably mobile in migratory bird populations where it has led to extensive outbreaks for which the true number of affected birds usually cannot be determined. For the evaluation of avian influenza monitoring and HPAIV early warning systems, we propose a time series analysis that includes the estimation of confidence intervals for (i) the prevalence in outbreak situations or (ii) in the apparent absence of disease in time intervals for specified regional units. For the German outbreak regions in 2006 and 2007, the upper 95% confidence limit allowed the detection of prevalences below 1% only for certain time intervals. Although more than 25,000 birds were sampled in Germany per year, the upper 95% confidence limit did not fall below 5% in the outbreak regions for most of the time. The proposed analysis can be used to monitor water bodies and high risk areas, also as part of an early warning system. Chances for an improved targeting of the monitoring system as part of a risk-based approach are discussed with the perspective of reducing sample sizes. PMID- 19680558 TI - Loss of anti-Bax function in Gerstmann-Straussler-Scheinker syndrome-associated prion protein mutants. AB - Previously, we have shown the loss of anti-Bax function in Creutzfeldt Jakob disease (CJD)-associated prion protein (PrP) mutants that are unable to generate cytosolic PrP (CyPrP). To determine if the anti-Bax function of PrP modulates the manifestation of prion diseases, we further investigated the anti-Bax function of eight familial Gerstmann-Straussler-Scheinker Syndrome (GSS)-associated PrP mutants. These PrP mutants contained their respective methionine ((M)) or valine ((V)) at codon 129. All of the mutants lost their ability to prevent Bax-mediated chromatin condensation or DNA fragmentation in primary human neurons. In the breast carcinoma MCF-7 cells, the F198S(V), D202N(V), P102L(V) and Q217R(V) retained, whereas the P102L(M), P105L(V), Y145stop(M) and Q212P(M) PrP mutants lost their ability to inhibit Bax-mediated condensed chromatin. The inhibition of Bax-mediated condensed chromatin depended on the ability of the mutants to generate cytosolic PrP. However, except for the P102L(V), none of the mutants significantly inhibited Bax-mediated caspase activation. These results show that the cytosolic PrP generated from the GSS mutants is not as efficient as wild type PrP in inhibiting Bax-mediated cell death. Furthermore, these results indicate that the anti-Bax function is also disrupted in GSS-associated PrP mutants and is not associated with the difference between CJD and GSS. PMID- 19680561 TI - Parkin deficiency delays motor decline and disease manifestation in a mouse model of synucleinopathy. AB - In synucleinopathies, including Parkinson's disease, partially ubiquitylated alpha-synuclein species phosphorylated on serine 129 (P(S129)-alpha-synuclein) accumulate abnormally. Parkin, an ubiquitin-protein ligase that is dysfunctional in autosomal recessive parkinsonism, protects against alpha-synuclein-mediated toxicity in various models.We analyzed the effects of Parkin deficiency in a mouse model of synucleinopathy to explore the possibility that Parkin and alpha synuclein act in the same biochemical pathway. Whether or not Parkin was present, these mice developed an age-dependent neurodegenerative disorder preceded by a progressive decline in performance in tasks predictive of sensorimotor dysfunction. The symptoms were accompanied by the deposition of P(S129)-alpha synuclein but not P(S87)-alpha-synuclein in neuronal cell bodies and neuritic processes throughout the brainstem and the spinal cord; activation of caspase 9 was observed in 5% of the P(S129)-alpha-synuclein-positive neurons. As in Lewy bodies, ubiquitin-immunoreactivity, albeit less abundant, was invariably co localized with P(S129)-alpha-synuclein. During late disease stages, the disease specific neuropathological features revealed by ubiquitin- and P(S129)-alpha synuclein-specific antibodies were similar in mice with or without Parkin. However, the proportion of P(S129)-alpha-synuclein-immunoreactive neuronal cell bodies and neurites co-stained for ubiquitin was lower in the absence than in the presence of Parkin, suggesting less advanced synucleinopathy. Moreover, sensorimotor impairment and manifestation of the neurodegenerative phenotype due to overproduction of human alpha-synuclein were significantly delayed in Parkin deficient mice.These findings raise the possibility that effective compensatory mechanisms modulate the phenotypic expression of disease in parkin-related parkinsonism. PMID- 19680567 TI - Special issue on fundamental principles and techniques in microfluidics. PMID- 19680569 TI - A brief introduction to slippage, droplets and mixing in microfluidic systems. PMID- 19680571 TI - Basic principles of electrolyte chemistry for microfluidic electrokinetics. Part II: Coupling between ion mobility, electrolysis, and acid-base equilibria. AB - We present elements of electrolyte dynamics and electrochemistry relevant to microfluidic electrokinetics experiments. In Part I of this two-paper series, we presented a review and introduction to the fundamentals of acid-base chemistry. Here, we first summarize the coupling between acid-base equilibrium chemistry and electrophoretic mobilities of electrolytes, at both infinite and finite dilution. We then discuss the effects of electrode reactions on microfluidic electrokinetic experiments and derive a model for pH changes in microchip reservoirs during typical direct-current electrokinetic experiments. We present a model for the potential drop in typical microchip electrophoresis device. The latter includes finite element simulation to estimate the relative effects of channel and reservoir dimensions. Finally, we summarize effects of electrode and electrolyte characteristics on potential drop in microfluidic devices. As a whole, the discussions highlight the importance of the coupling between electromigration and electrophoresis, acid-base equilibria, and electrochemical reactions. PMID- 19680570 TI - Basic principles of electrolyte chemistry for microfluidic electrokinetics. Part I: Acid-base equilibria and pH buffers. AB - We review fundamental and applied acid-base equilibrium chemistry useful to microfluidic electrokinetics. We present elements of acid-base equilibrium reactions and derive rules for pH calculation for simple buffers. We also present a general formulation to calculate pH of more complex, arbitrary mixtures of electrolytes, and discuss the effects of ionic strength and temperature on pH calculation. More practically, we offer advice on buffer preparation and on buffer reporting. We also discuss "real world" buffers and likely contamination sources. In particular, we discuss the effects of atmospheric carbon dioxide on buffer systems, namely, the increase in ionic strength and acidification of typical electrokinetic device buffers. In Part II of this two-paper series, we discuss the coupling of acid-base equilibria with electrolyte dynamics and electrochemistry in typical microfluidic electrokinetic systems. PMID- 19680562 TI - A "repertoire for repertoire" hypothesis: repertoires of type three effectors are candidate determinants of host specificity in Xanthomonas. AB - BACKGROUND: The genetic basis of host specificity for animal and plant pathogenic bacteria remains poorly understood. For plant pathogenic bacteria, host range is restricted to one or a few host plant species reflecting a tight adaptation to specific hosts. METHODOLOGY/PRINCIPAL FINDINGS: Two hypotheses can be formulated to explain host specificity: either it can be explained by the phylogenetic position of the strains, or by the association of virulence genes enabling a pathological convergence of phylogenically distant strains. In this latter hypothesis, host specificity would result from the interaction between repertoires of bacterial virulence genes and repertoires of genes involved in host defences. To challenge these two hypotheses, we selected 132 Xanthomonas axonopodis strains representative of 18 different pathovars which display different host range. First, the phylogenetic position of each strain was determined by sequencing the housekeeping gene rpoD. This study showed that many pathovars of Xanthomonas axonopodis are polyphyletic. Second, we investigated the distribution of 35 type III effector genes (T3Es) in these strains by both PCR and hybridization methods. Indeed, for pathogenic bacteria T3Es were shown to trigger and to subvert host defences. Our study revealed that T3E repertoires comprise core and variable gene suites that likely have distinct roles in pathogenicity and different evolutionary histories. Our results showed a correspondence between composition of T3E repertoires and pathovars of Xanthomonas axonopodis. For polyphyletic pathovars, this suggests that T3E genes might explain a pathological convergence of phylogenetically distant strains. We also identified several DNA rearrangements within T3E genes, some of which correlate with host specificity of strains. CONCLUSIONS/SIGNIFICANCE: These data provide insight into the potential role played by T3E genes for pathogenic bacteria and support a "repertoire for repertoire" hypothesis that may explain host specificity. Our work provides resources for functional and evolutionary studies aiming at understanding host specificity of pathogenic bacteria, functional redundancy between T3Es and the driving forces shaping T3E repertoires. PMID- 19680572 TI - Parallel multiphase microflows: fundamental physics, stabilization methods and applications. AB - Parallel multiphase microflows, which can integrate unit operations in a microchip under continuous flow conditions, are discussed. Fundamental physics, stabilization methods and some applications are shown. PMID- 19680573 TI - Induced-charge electrokinetics: fundamental challenges and opportunities. AB - Induced-charge electrokinetic (ICEK) phenomena occur when an applied electric field induces an ionic double-layer over a polarizable surface, then forces that induced double-layer into electro-osmotic flow. The nonlinear character of ICEK phenomena enable steady flows to be driven using small AC potentials, which in turn reduces or eliminates electrochemical reactions. As such, ICEK holds promise as a mechanism by which low-voltage, high-pressure pumps may be developed to enable portable, self-contained microfluidic manipulation. Here we review the basic physics of induced-charge electrokinetic phenomena and the advantages they hold for Lab-on-a-Chip devices, in addition to the opportunities they present for fundamental science. In particular, these systems are unique in that all aspects of the (measurable) ICEK flows can be predicted in advance using standard electrokinetic theories, which can then be compared with experimental data. Such comparisons have revealed a number of striking discrepancies between theory and experiment, thus indicating the standard model is missing key physical or chemical ingredients. We discuss specific discrepancies-which remain to be understood-and the challenges they pose for widespread implementation of ICEK in practical Lab-on-a-Chip devices. We propose and present a variety of challenges and opportunities-theoretical and experimental, fundamental and applied-which must be addressed. PMID- 19680575 TI - Microchemical systems for continuous-flow synthesis. AB - Microchemical systems have evolved rapidly over the last decade with extensive chemistry applications. Such systems enable discovery and development of synthetic routes while simultaneously providing increased understanding of underlying pathways and kinetics. We review basic trends and aspects of microsystems as they relate to continuous-flow microchemical synthesis. Key literature reviews are summarized and principles governing different microchemical operations discussed. Current trends and limitations of microfabrication, micromixing, chemical synthesis in microreactors, continuous flow separations, multi-step synthesis, and integration of analytics are delineated. We conclude by summarizing the major challenges and outlook related to these topics. PMID- 19680574 TI - Nucleic acid extraction techniques and application to the microchip. AB - As recently as the early 1990s, DNA purification was time-consuming, requiring the use of toxic, hazardous reagents. The advent of solid phase extraction techniques and the availability of commercial kits for quick and reliable DNA extraction has relegated those early techniques largely to the history books. High quality DNA can now be extracted from whole blood, serum, saliva, urine, stool, cerebral spinal fluid, tissues, and cells in less time without sacrificing recovery. Having achieved such a radical change in the methodology of DNA extraction, focus has shifted to adapting these methods to a miniaturized system, or "lab-on-a-chip" (A. Manz, N. Graber and H. M. Widmer, Sens. Actuators, B, 1990, 1, 244-248). Manz et al.'s concept of a "miniaturized total chemical analysis system" (microTAS) involved a silicon chip that incorporated sample pretreatment, separation and detection. This review will focus on the first of these steps, sample pretreatment in the form of DNA purification. The intention of this review is to provide an overview of the fundamentals of nucleic acid purification and solid phase extraction (SPE) and to discuss specific microchip DNA extraction successes and challenges. In order to fully appreciate the advances in DNA purification, a brief review of the history of DNA extraction is provided so that the reader has an understanding of the impact that the development of SPE techniques have had. This review will highlight the different methods of nucleic acid extraction (Table 1), including relevant citations, but without an exhaustive summary of the literature. A recent review by Wen et al. (J. Wen, L. A. Legendre, J. M. Bienvenue and J. P. Landers, Anal. Chem., 2008, 80, 6472-6479) covers solid phase extraction methods with a greater focus on their incorporation into integrated microfluidic systems. PMID- 19680576 TI - Electrophoretic separation of DNA in gels and nanostructures. AB - The development of nanostructure devices has opened the door to new DNA separation techniques and fundamental investigations. With advanced nanotechnologies, artificial gels (e.g. nanopillar arrays, nanofilters) can be manufactured with controlled and ordered geometries. This contrast with gels, where the pores are disordered and the range of available pore sizes is limited by the level of cross-linking and the mechanical properties of the gel. In this review, we recall the theories developed for free-solution and gel electrophoresis (extended Ogston model, biased reptation and entropic trapping) and from this perspective, suggestions for future concepts for fast DNA separation using nanostructures will be given. PMID- 19680577 TI - On-chip technologies for multidimensional separations. AB - We review microfluidic devices designed for multidimensional sample analysis, with a primer on relevant theory, an emphasis on protein analysis, and an eye towards future improvements and challenges to the field. Image shows results of an on-chip IEF-CE separation of a protein mixture; unpublished surface plot data from A. E. Herr. PMID- 19680578 TI - Characterizing dispersion in microfluidic channels. AB - Dispersion or spreading of analyte bands is a barrier to achieving high resolution in microfluidic separations. The role of dispersion in separations is reviewed with emphasis on metrics, sources and common principles of analysis. Three sources of dispersion (a) inhomogeneous flow fields, (b) solute wall interactions and (c) force fields normal to channel walls are studied in detail. Microfluidic and nanofluidic applications to capillary electrophoresis, chromatography and field-flow fractionation, that are subject to one or more of these three physical processes under standard, unintentional or novel operating conditions, are discussed. PMID- 19680579 TI - Micro-Particle Image Velocimetry (microPIV): recent developments, applications, and guidelines. AB - In this review we discuss the state of the art of the optical whole-field velocity measurement technique micro-scale Particle Image Velocimetry (microPIV). microPIV is a useful tool for fundamental research of microfluidics as well as for the detailed characterization and optimization of microfluidic applications in life science, lab-on-a-chip, biomedical research, micro chemical engineering, analytical chemistry and other related fields of research. An in depth description of the microPIV method is presented and compared to other flow visualization and measurement methods. An overview of the most relevant applications is given on the topics of near-wall flow, electrokinetic flow, biological flow, mixing, two-phase flow, turbulence transition and complex fluid dynamic problems. Current trends and applications are critically reviewed. Guidelines for the implementation and application are also discussed. PMID- 19680580 TI - Microrheology with optical tweezers. AB - Microrheology is the study of the flow of materials over small scales. It is of particular interest to those involved with investigations of fluid properties within Lab-on-a-Chip structures or within other micron-scale environments. The article briefly reviews existing active and passive methods used in the study of fluids. It then explores in greater detail the use of optical tweezers as an emerging method to investigate rheological phenomena, including, for example, viscosity and viscoelasticity, as well as the related topic of flow. The article also describes, briefly, potential future applications of this topic, in the fields of biological measurement, in general, and Lab-on-a-Chip, in particular. PMID- 19680581 TI - Neuro-optical microfluidic platform to study injury and regeneration of single axons. AB - We describe a neuro-optical microfluidic platform for studying injury and subsequent regeneration of individual mammalian axons. This platform consists of three components integrated on an inverted microscope, which include a compartmentalized neuronal culture microfluidic device, a femtosecond laser to enable precise axotomy, and a custom built mini cell culture incubator for continuous long term observation of post injury events. We demonstrate the unique capabilities of the platform by injuring individual central and peripheral nervous system axons and monitoring the post injury sequence of events from initial degeneration to subsequent regeneration. This platform will enable study and understanding of neuronal response to injury that is currently not possible with conventional cell culture platform and tools. PMID- 19680582 TI - A microfluidic-based method for the transfer of biopolymer particles from an oil phase to an aqueous phase. AB - Biopolymer microgels produced in microfluidic devices via the formation of a water-in-oil emulsion are usually collected at the outlet of the device and thoroughly washed from the oil phase in an additional, lengthy processing step. This paper reports a microfluidic-based method which allows for continuous on chip manufacture of aqueous-based biopolymer microparticles in an oily continuous phase and thereafter the transfer of these particles from the oily carrier phase to a second aqueous continuous phase. This was achieved by surface patterning the PDMS channel walls using UV polymerization of poly(acrylic acid) (PAA) in order to obtain a hybrid device with distinct hydrophilic and hydrophobic sections. The surface patterning was stable for at least 4 months. This selective surface patterning of the channel was shown to initiate and assist the transfer of the biopolymer particles from the oil phase into the aqueous phase. The flow conditions required for a stable biphasic flow in the transfer section of the device were evaluated based on the theoretical shear stress at the interface of the two fluids. Experimental outcomes were found to be in good agreement with the prediction. After the particles cross the liquid-liquid interface and are transferred into the aqueous phase, they are collected and characterized. The resulting suspension was found to be stable for several weeks and no aggregation was observed. PMID- 19680583 TI - Microfluidic platform for controlling the differentiation of embryoid bodies. AB - Embryonic stem (ES) cells are pluripotent cells, which can differentiate into any cell type. This cell type has often been implicated as an eminent source of renewable cells for tissue regeneration and cellular replacement therapies. Studies on manipulation of the various differentiation pathways have been at the forefront of research. There are many ways in which ES cells can be differentiated. One of the most common techniques is to initiate the development of embryoid bodies (EBs) by in vitro aggregation of ES cells. Thereafter, EBs can be induced to undergo differentiation into various cell lineages. In this article, we present a microfluidic platform using biocompatible materials, which is suitable for culturing EBs. The platform is based on a Y-channel device with two inlets for two different culturing media. An EB is located across both streams. Using the laminar characteristics at low Reynolds number and high Peclet numbers, we have induced cell differentiation on half of the EB while maintaining the other half in un-induced stages. The results prove the potential of using microfluidic technology for manipulation of EBs and ES cells in tissue engineering. PMID- 19680584 TI - Microfluidic synthesis of a cell adhesive Janus polyurethane microfiber. AB - We present a simple synthetic approach for the preparation of cell attachable Janus polyurethane (PU) microfibers in a microfluidic system. The synthesis was performed by using laminar flows of multiple streams with spontaneous formation of carbon dioxide bubbles resulting in an asymmetrically porous PU microfiber. The fabricated asymmetric microfiber (Janus microfiber) provides two distinctive properties: one is a porous region to promote the cellular adhesion and the other is a nonporous region rendering the mechanical strength of the scaffold. The Janus microfibers show dramatic improvement of cell adhesion, proliferation, and viability over a culture period. Cells cultured on the fibers easily bridged gaps between microfibers by joining together to form a cell sheet. The maximum distance between fibers that fibroblasts bridged is approximately 200 microm over 15 days. The Janus microfiber can be used for not only an alternative 2D cell culture plate but also as a novel 3D scaffold for tissue engineering without any need for elegant surface modification for enhancing cell adhesions. PMID- 19680585 TI - Modulation of fluidic resistance and capacitance for long-term, high-speed feedback control of a microfluidic interface. AB - Existing microfluidic systems can control local chemical environments by directing the interface between laminar flowing streams for applications ranging from subcellular stimulation to fuel cells. However, conventional flow modulation methods have not yet provided a robust and reliable way to dynamically control laminar flow interfaces for very long time periods. Such control is important in biological investigations, since response times for living cells and tissues can be as long as several days. Here, we describe a novel long-term, high-speed approach that employs modulation of fluidic resistance and fluidic capacitance between a fluid reservoir and a microfluidic network with feedback control to enable long-term dynamic control of a microfluidic interface in time and space. Our method involves constricting a narrow tube through a pinching approach to modulate fluidic resistance while also controlling a small variable reservoir in the fluidic network through a squeezing approach to modulate fluidic capacitance. We designed a well-tuned proportional-integral-derivative (PID) controller for the closed-loop control system that resulted in control of pressure for short term (2 s) and long-term (15 h) experiments. Further, we integrated a pressure based feedback control approach into this method, which enables both long-term spatiotemporal control of our microfluidic interface at frequencies greater than 1 Hz and a reservoir capacity to enable experiments for longer than 60 days. This long-term and high-speed control is not possible with standard microfluidic laboratory practices. Our system has a diversity of potential applications including long-term cellular studies in cancer metastasis or embryonic development. PMID- 19680588 TI - Mental illness stigma and willingness to seek mental health care in the European Union. AB - OBJECTIVE: It is often assumed that individual stigmatizing attitudes toward the mentally ill are linked to stigmatizing attitudes in the social milieu and that both, individual and social stigmatizing attitudes are major barriers to mental health treatment seeking. This study aims to examine these assumptions. METHOD: Data from the 2005-2006 Eurobarometer general population survey (N = 29,248) are used to examine the association of social stigmatizing attitudes assessed in a random half of the sample with individual stigmatizing attitudes assessed in the other half of the sample, and to examine the association of both individual and social stigmatizing attitudes with willingness to seek professional help. RESULTS: Social stigmatizing attitudes are specifically and strongly associated with individual stigmatizing attitudes. Both social and individual stigmatizing attitudes are associated with willingness to seek professional help. Believing the mentally ill to be dangerous or not likely to recover, or living in a community with such beliefs, are associated with increased willingness to seek help; whereas, believing the mentally ill to be unpredictable or blameworthy for their illness, or living in a community with strong beliefs in blameworthiness of the mentally ill, are associated with decreased willingness to seek professional help. CONCLUSION: The view that all stigmatizing attitudes toward mental illness are associated with reluctance to seek professional help may be naive as some stigmatizing attitudes may be associated with increased willingness to seek help. The complex association of different stigmatizing attitudes with professional help seeking should be carefully considered in planning anti-stigma campaigns. PMID- 19680590 TI - Relationship between historical height loss and vertebral fractures in postmenopausal women. AB - The aim of this study was to evaluate the relationship between historical height loss (HHL) and prevalent vertebral fractures (VF) in postmenopausal Moroccan women and to estimate its accuracy as a clinical test for detecting VF. Two hundred eighty-eight postmenopausal women were studied. All subjects had bone density measurements and spinal radiographs. Vertebral bodies (T4-L4) were graded using the semi-quantitative method of Genant. HHL was calculated as the difference between a patient's tallest recalled height and the current measured height. The mean age was 58.4 +/- 7.8 years. Thirty-one percent of patients were osteoporotic, and 46.5% had VF. Patients with VF had lost more height than those without VF (median, 2.0 cm (0.26-3.3) vs 0.96 cm (0.33-2.4), p < 0.05). In univariate analysis, HHL was positively correlated to both number and grade of prevalent VF (p < 0.05). The area under the receiver operating characteristics curve for the ability of HHL to detect VF was 0.60 (95% confidence interval (CI), 0.52, 0.69). Our HHL threshold for detecting VF was >1.5 cm, its sensitivity was 58%, and its specificity was 61%. The positive predictive value was 53%, and the negative predictive value was 65%. With HHL >1.5 cm, positive likelihood ratio was 1.49 with 95% CI, 1.07, 2.06. Our results demonstrate significant positive associations between HHL, VF, number of VF, and grade of VF. However, this relationship is not clinically pertinent. Consequently, HHL cannot be used as a reliable clinical test for detecting VF in postmenopausal Moroccan women. PMID- 19680591 TI - Occurrence of antifouling biocides and fluorinated alkyl compounds in sediment core from deep sea: Suruga Bay, Tosa Bay, and Nankai tough, Japan. AB - Contamination profiles of antifouling biocides were investigated in a deep-sea environment in Suruga Bay Japan. Significant differences in the tributyltin (TBT) and triphenyltin (TPT) concentrations in subsurface sediment between 850 and 800 m of water depth were not observed (p < 0.05). Organotin (OT) concentrations in sediment core of 0-30.5 cm from a water depth of 800 m were investigated. The butyltins (BTs) and phenyltins (PTs) concentrations were constant between 0 and 15 cm, and, subsequently, the concentration of these compounds increased. The peaks of the BTs and PTs concentrations were observed between 18 and 19 cm. The concentrations of Irgarol 1051 decreased until a core depth of 9 cm, and, the values then became near the detection limit under the 10 cm of core depth. Perfluorooctane sulfonate (PFOS) and perfluorooctanic acid (PFOA) were detected in the sediment core (0-30.5 cm) of Suruga Bay. The concentration of PFOS was high in the 0-5-cm core depth and then decreased. The concentrations of PFOA, however, were at the values near the detection limit throughout the sediment core. The BTs and PTs concentrations in surface sediment from Tosa Bay decreased with water depth. Although Irgarol 1051 was the only alternative compound detected, the value was near the detection limit. PFOS and PFOA were detected in sediment core from Tosa Bay. The concentrations of PFOS became low as the water depth became deeper. TBT, TPT, Sea Nine 211, Diuron and Irgarol 1051 were detected in sediment core (core depth: 10 cm) from the Nankai trough (water depth: 4010 m). PMID- 19680593 TI - Role of p38 and JNK in liver ischemia and reperfusion. AB - BACKGROUND/PURPOSE: The signal transduction of mitogen-activated protein kinases (MAPKs) has appeared to be an important mediator of ischemic-related events. Because of this, we analyzed the participation of p38 and JNK in liver ischemia and reperfusion, as two individual members of the MAPK family of proteins. METHODS: All papers referred to in PubMed for the past 15 years were analyzed to determine how and when these MAPKs were considered to be an intricate part of the ischemic event. References were cross-studied to ascertain whether other papers could be found in the literature. RESULTS: The role of p38 and JNK in liver ischemia was confirmed in the literature. The activation of these mediators was associated with the induction of apoptosis and necrosis. Inhibitors of p38 and JNK reduced the liver ischemia and reperfusion damage, probably through the mechanisms mentioned before. CONCLUSIONS: The development of effective inhibitors of p38 and JNK protein mediators is important for minimizing the harmful effects associated with liver ischemia and reperfusion. PMID- 19680589 TI - Genome-wide copy number variation association study suggested VPS13B gene for osteoporosis in Caucasians. AB - Osteoporotic fracture (OF) is a serious outcome of osteoporosis. Important risk factors for OF include reduced bone mineral density and unstable bone structure. This genome-wide copy number variation association study suggested VPS13B gene for osteoporosis in Caucasians. INTRODUCTION: Bone mineral density (BMD) and femoral neck cross-sectional geometric parameters (FNCSGPs) are under strong genetic control. DNA copy number variation (CNV) is an important source of genetic diversity for human diseases. This study aims to identify CNVs associated with BMD and FNCSGPs. METHODS: Genome-wide CNV association analyses were conducted in 1,000 unrelated Caucasian subjects for BMD at the spine, hip, femoral neck, and for three FNCSGPs -cortical thickness (CT), cross-section area (CSA), and buckling ratio (BR). BMD was measured by dual energy X-ray absorptiometry (DEXA). CT, CSA, and BR were estimated using DEXA measurements. Affymetrix 500K arrays and copy number analysis tool was used to identify CNVs. RESULTS: A CNV in VPS13B gene was significantly associated with spine, hip and FN BMDs, and CT, CSA, and BR (p < 0.05). Compared to subjects with two copies of the CNV, carriers of one copy had an average of 14.6%, 12.4%, and 13.6% higher spine, hip, and FN BMD, 20.0% thicker CT, 10.6% larger CSA, and 12.4% lower BR. Thus, a decrease of the CNV consistently produced stronger bone, thereby reducing osteoporotic fracture risk. CONCLUSIONS: VPS13B gene, via affecting BMD and FNCSGPs, is a novel osteoporosis risk gene. PMID- 19680592 TI - Tumorigenesis and phenotypic characteristics of mucin-producing bile duct tumors: an immunohistochemical approach. AB - Intraductal papillary neoplasm of the bile duct (IPNB) is characterized by exophytic proliferation of neoplastic epithelial cells with fibrovascular stalks in bile duct lumen, mucin hypersecretion, and considerable dilatation or multilocular changes of the affected bile ducts. A mucin-producing bile duct tumor is an IPNB with excessive mucin production and clinical symptoms. Herein, the phenotypes as well as the tumorigenesis and progression of IPNB are reviewed with immunohistochemical assistance. The tumors are subdivided into three phenotypes: pancreatobiliary, intestinal, and gastric. About half of IPNB cases are of the pancreatobiliary type, and the remaining half are of the intestinal type. Aberrant expression of CDX2 with MUC2 and CK20 is related to the development of intestinal metaplasia. Inactivation of P16INK4a and nuclear expression of beta-catenin are related to the development of IPNB. Decreased expression of membranous beta-catenin and E-cadherin and aberrant expression of MMP-7 and -9 and of MUC1 are related to invasion of IPNB with tubular adenocarcinoma, whereas MUC2 is involved in the invasion of IPNB with mucinous carcinoma. IPNB can be regarded as a counterpart of intraductal papillary mucinous neoplasm (IPMN) of the pancreas, particularly the main duct type. More comparative studies between IPNB and pancreatic IPMN are recommended for further analysis of these papillary neoplasms. PMID- 19680594 TI - Clinical assessment of atherosclerotic parameters and cardiac function in chronic hemodialysis patients. AB - BACKGROUND/AIM: Atherosclerosis is evaluated by carotid mean intima-media thickness (mean IMT), pulse wave velocity (PWV), and the aortic calcification index (ACI). We have attempted to examine if these atherosclerotic parameters are associated with each other and which parameters are closely related to cardiac function in chronic HD patients. METHODS: The mean IMT, PWV and ACI were examined in 69 maintenance HD patients using carotid ultrasonography, a blood volume plethysmographic apparatus and abdominal CT, respectively. Echocardiographic studies were also performed for measuring left ventricular (LV) geometry. Serum total cholesterol, high density lipoprotein cholesterol, low density lipoprotein cholesterol, triglyceride, albumin, C-reactive protein (CRP), calcium and phosphate were measured. RESULTS: The mean IMT correlated positively with ACI (r = 0.461, P < 0.0001) and tended to be correlated with PWV, but did not reach statistical significance. The PWV value correlated positively with ACI (r = 0.494, P 0.3 during CATIE phase 1, N = 199) or controls (average SAS mean global score 0, N = 198). RESULTS: Using logistic regression and controlling for population stratification, age, gender, SAS score at baseline, and concomitant use of anticholinergic drugs, we identified several single-nucleotide polymorphisms associated with AIP severity. Although none reached the GWAS significance level of P < 4.2 x 10(-7), some promising candidate genes for further research on genetic predisposition to AIP were identified including EPF1, NOVA1, and FIGN. CONCLUSIONS: Our finding may contribute to understanding of the pathophysiology of AIP as well as to a priori identification of patients vulnerable for development of AIP. PMID- 19680637 TI - Direct immobilization of functional single-chain variable fragment antibodies (scFvs) onto a polystyrene plate by genetic fusion of a polystyrene-binding peptide (PS-tag). AB - Single-chain Fv antibodies (scFv) genetically fused with polystyrene-binding peptides (PS-tags, (PS19-1; RAFIASRRIRRP, PS19-6; RIIIRRIRR)) were generated by recombinant Escherichia coli for direct and site-specific immobilization of scFv on polystyrene supports with high antigen-binding activity. PS-tag-fused scFvs (scFv-PS-tags) specific for human C-reactive protein (CRP) were successfully over expressed as an inclusion body and were refolded using the batch-dilution method. When scFv-PS-tags were immobilized on a hydrophilic PS (phi-PS) plate in the presence of Tween 20, they showed high antigen-binding activity comparable to, or greater than, that of a whole monoclonal antibody (mAb) on a hydrophobic PS (pho PS) plate, which has been the exclusive method for enzyme-linked immunosorbent assay (ELISA). Furthermore, when a scFv-PS-tag was used as a ligand antibody in one- and two-step ELISA, the assay time was reduced without loss of sensitivity. These results indicate that strong and specific attachment of PS-tags onto the phi-PS surface prevented scFv conformational changes and consequently, the high antigen-binding activities of scFvs were preserved. Nearly identical results were obtained by use of PS-tag-fused scFvs with different VH/VL pairs. Therefore, a variety of scFvs could be functionalized onto phi-PS plates by genetic fusion of PS-tags. ScFv-PS-tags, which possess high antigen-binding activity on the phi-PS plate, are more useful ligand antibodies than whole mAbs. Thus, scFv-PS-tags are applicable in both clinical diagnosis and proteomic research. PMID- 19680638 TI - DC- and RF-GD-OES measurements of adsorbed organic monolayers on copper. AB - Our direct current (DC)- and radiofrequency glow discharge optical emission spectroscopy (RF-GD-OES) measurements of adsorbed organic monolayers were inspired by the work of Shimizu et al., who presented the first example of depth profile analysis of an adsorbed monolayer by RF-GD-OES in 2004. The great potential of RF-GD-OES for analyses of layers with thicknesses in the subnanometer range was surprising. Shimizu et al. discussed not only the qualitative detection of atoms of the organic monolayer (C, H, N, S), but also the determination of the different orientation of the molecules relative to the surface due to a significant peak sequence. This latter assumption was questioned in the analytical community. We intend to demonstrate the potential of the GD-OES technique for surface analysis in terms of reliability and reproducibility by using an advanced vacuum instrumentation and presputtering with silicon. It will be shown that comparable measurements can be reproduced not only with RF-GD-OES but, above all, also with DC-GD-OES. The experimental steps to adsorb thiourea molecules on a copper substrate are described in detail. Further experiments with other organic molecules, e.g. benzotriazole (BTA) or benzothiazole (BTH), disprove the predicted correlation between the orientation of the molecules relative to the surface and the occurrence of peak separation. Ultimately, a quantification of compounds of the organic monolayer in the case of adsorbed thiourea is achieved. PMID- 19680639 TI - Voluntary action and causality in temporal binding. AB - Previous studies have documented temporal attraction in perceived times of actions and their effects. While some authors argue that voluntary action is a necessary condition for this phenomenon, others claim that the causal relationship between action and effect is the crucial ingredient. In the present study, we investigate voluntary action and causality as the necessary and sufficient conditions for temporal binding. We used a variation of the launching effect proposed by Michotte, in which participants controlled the launch stimulus in some blocks. Volunteers reported causality ratings and estimated the interval between the two events. Our results show dissociations between causality ratings and temporal estimation. While causality ratings are not affected by voluntary action, temporal bindings were only found in the presence of both voluntary action and high causality. Our results indicate that voluntary action and causality are both necessary for the emergence of temporal binding. PMID- 19680642 TI - Torsion of testicular appendage. PMID- 19680643 TI - Clinical image. The "wandering" spleen. PMID- 19680640 TI - Effects of visual gain on force control at the elbow and ankle. AB - Visual feedback is essential when minimizing force fluctuations. Varying degrees of somatotopic organization have been shown in different regions of the brain for the upper and lower extremities, and visual feedback may be processed differently based on the body effector where feedback-based corrections are used. This study compared the effect of changes in visual gain on the control of steady-state force at the elbow and ankle. Ten subjects produced steady-state isometric force to targets at 5 and 40% of their maximum voluntary contraction at seven visual gain levels. Visual gain was used effectively at both joints to reduce variability of the force signal and to improve accuracy, with a greater effect of visual gain at the elbow than the ankle. Visual gain significantly decreased the regularity of force output, and this effect was more pronounced at the elbow than the ankle. There were accompanying changes in the proportion of power in the 0-4, 4-8, and 8-12 Hz frequency bins of the force signal across visual gain at the elbow. Changes in visual gain were accompanied by changes in both agonist and antagonist electromyographic (EMG) activation at the elbow. At the ankle joint, there were only changes in agonist EMG. The results suggest better use of visual information in the control of elbow force than ankle force and this improved control may be related to the changes in the pattern of agonist and antagonist activation. PMID- 19680644 TI - Viruses: incredible nanomachines. New advances with filamentous phages. AB - During recent decades, bacteriophages have been at the cutting edge of new developments in molecular biology, biophysics, and, more recently, bionanotechnology. In particular filamentous viruses, for example bacteriophage M13, have a virion architecture that enables precision building of ordered and defect-free two and three-dimensional structures on a nanometre scale. This could not have been possible without detailed knowledge of coat protein structure and dynamics during the virus reproduction cycle. The results of the spectroscopic studies conducted in our group compellingly demonstrate a critical role of membrane embedment of the protein both during infectious entry of the virus into the host cell and during assembly of the new virion in the host membrane. The protein is effectively embedded in the membrane by a strong C-terminal interfacial anchor, which together with a simple tilt mechanism and a subtle structural adjustment of the extreme end of its N terminus provides favourable thermodynamical association of the protein in the lipid bilayer. This basic physicochemical rule cannot be violated and any new bionanotechnology that will emerge from bacteriophage M13 should take this into account. PMID- 19680646 TI - Simple adaptive pH control in bioreactors using gain-scheduling methods. AB - A simple well-performing adaptive control technique for pH control in fermentations of recombinant protein production processes is described and its design procedure is explained. First, the entire control algorithm was simulated and parameterized. Afterwards it was tested in real cultivation processes. The results show that this simple technique leads to significant reductions in the fluctuations of the pH values in microbial cultures at a minimum of expenditures. The signal-to-noise ratio and thus the information captured by the pH signal were increased by about an order of magnitude. This leads to a substantial improvement in the noise of many other process signals that are used to monitor and control the process. For instance, respiratory off-gas data of CO(2) and its derived carbon dioxide production rate signals from the cultures carry much less noise as compared to those values obtained with conventional pH control. Detailed process analysis revealed that even very small pH jumps of 0.03 values during the fermentation were shown to result in pronounced deflections in CO(2)-volume fraction of 8% (peak to peak). The proposed controller, maintaining the pH within the interval of 0.01 around the setpoint, reduces the noise considerably. PMID- 19680645 TI - In vivo magnetic resonance spectroscopy: basic methodology and clinical applications. AB - The clinical use of in vivo magnetic resonance spectroscopy (MRS) has been limited for a long time, mainly due to its low sensitivity. However, with the advent of clinical MR systems with higher magnetic field strengths such as 3 Tesla, the development of better coils, and the design of optimized radio frequency pulses, sensitivity has been considerably improved. Therefore, in vivo MRS has become a technique that is routinely used more and more in the clinic. In this review, the basic methodology of in vivo MRS is described-mainly focused on (1)H MRS of the brain-with attention to hardware requirements, patient safety, acquisition methods, data post-processing, and quantification. Furthermore, examples of clinical applications of in vivo brain MRS in two interesting fields are described. First, together with a description of the major resonances present in brain MR spectra, several examples are presented of deviations from the normal spectral pattern associated with inborn errors of metabolism. Second, through examples of MR spectra of brain tumors, it is shown that MRS can play an important role in oncology. PMID- 19680647 TI - Skeletal metastases: what is the future role for nuclear medicine? PMID- 19680649 TI - Autophagy facilitates major histocompatibility complex class I expression induced by IFN-gamma in B16 melanoma cells. AB - The reduction or loss of MHC-I antigen surface expression in human and murine tumor cells is partly attributable to the dysregulation of various components of the MHC-I antigen-processing machinery. Accumulating evidence suggests that autophagy, besides its vital role in maintaining the cellular homeostasis, plays an important role in MHC-II surface expression. Here, we report that autophagy is a negative regulator of MHC-I antigen expression in B16 melanoma cells; however, in the presence of IFN-gamma, it is converted to a positive regulator. We show that autophagy not only participates in the degradation of MHC-I antigen but also plays a role in the generation of MHC-I-binding peptides. For these two processes, IFN-gamma interferes with MHC-I antigen degradation, rather than affecting peptide generation. Using B16 melanoma mouse model, we further show that autophagy may enhance the cytolysis of CTL to melanoma cells at the early stage of melanoma, but impairs the cytolysis at the late stage. Such different consequences may be explained by the different levels of IFN-gamma during tumor progression. Taken together, our findings demonstrate that autophagy is involved in the regulation of MHC-I antigen expression, through which autophagy can play different roles in tumor immunity. PMID- 19680650 TI - Antibody to CCDC104 is associated with a paraneoplastic antibody to CDR2 (anti Yo). AB - Patients with cancer may develop paraneoplastic neurological syndromes (PNS) in which onconeural antibodies are important diagnostic findings. As the functional role of onconeural antibodies is largely unknown, insight gained by identifying associated antibodies may help to clarify the pathogenesis of the PNS. In this study, we identified patients with Yo antibodies who also had antibodies to an uncharacterized protein called coiled-coil domain-containing protein 104 (CCDC104). We found a significant association between CCDC104 and Yo antibodies (4 of 38, 10.5%), but not other onconeural antibodies (0 of 158) (P = 0.007, Fisher's exact test). The prevalence of CCDC104 antibodies was approximately similar in patients with cancer (8 of 756, 1.1%) and in healthy blood donors (2 of 300, 0.7%). CCDC104 antibodies were not associated with PNS, as this was found in only two of the ten CCDC104-positive patients. The CCDC104 protein, whose function is unknown, is expressed in various human tissues, including the brain, and is localized mainly to the nucleus, but is also found in the cytoplasm. The association between Yo and CCDC104 antibodies may indicate functional similarities. PMID- 19680651 TI - Impact of surgical approach on postoperative heterotopic ossification and avascular necrosis in femoral head fractures: a systematic review. AB - Heterotopic ossification (HO) and avascular necrosis (AVN) have been identified as post-traumatic complications of femoral head fractures and may lead to a restriction in hip function and permanent disability. The question of which surgical approach is the best for the femoral head fracture and its relationship with HO and AVN remains controversial. We conducted a systematic review in which all published studies were evaluated. We performed a literature search in MEDLINE, PubMed, EMBASE, MD Consult, and the Cochrane Controlled Trial Register from 1980 to April 2009. We found ten appropriate studies, describing 176 patients. A lower percentage of patients treated with a trochanteric flip approach was reported with HO than patients treated with anterior or posterior approach (33.3% versus 42.1% and 36.9%, respectively), although the difference was not statistically significant. The incidence of AVN was highest in the posterior approach group (16.9%), and subsequently with the trochanteric flip approach (12.5%) and the anterior group (7.9%). The investigators concluded that the use of the anterior approach may result in a higher risk for HO and the posterior approach may result in a higher risk for AVN. A new, posterior-based approach of trochanteric flip seems to be a better approach for femoral head fractures. A further case-control study would be appropriate to confirm the findings in our systematic review. PMID- 19680652 TI - Erlotinib as salvage treatment after failure to first-line gefitinib in non-small cell lung cancer. AB - PURPOSE: Chemotherapy is the mainstay treatment for advanced non-small cell lung cancer (NSCLC). Gefitinib, an epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI), has been recently shown to be effective as a first-line treatment in Asian patients with advanced NSCLC, especially for those with favourable clinical features such as female, non-smoker and adenocarcinoma. However, resistance to gefitinib ensues invariably and there is little evidence as for the effectiveness of subsequent salvage treatment. The purpose of this study is to evaluate the efficacy of erlotinib, another EGFR-TKI, after failed first-line use of gefitinib. METHOD: Retrospective review of NSCLC patients with favourable clinical features who received gefitinib as first-line treatment and subsequent salvage treatment with erlotinib. RESULTS: A total of 21 patients with NSCLC were included in the study. Among them, 18 (85.7%) patients had disease control with gefitinib and 12 (57.1%) patients with salvage erlotinib. There was an association between the disease control with gefitinib and erlotinib (p = 0.031). The disease control rate of erlotinib was independent of the chemotherapy use between the two EGFR-TKIs. CONCLUSION: For NSCLC patients with favourable clinical features, erlotinib was effective in those who had prior disease control with first-line gefitinib. PMID- 19680653 TI - Induction of necrosis and cell cycle arrest in murine cancer cell lines by Melaleuca alternifolia (tea tree) oil and terpinen-4-ol. AB - PURPOSE: To examine the in vitro anticancer activity of Melaleuca alternifolia (tea tree) oil (TTO), and its major active terpene component, terpinen-4-ol, against two aggressive murine tumour cell lines, AE17 mesothelioma and B16 melanoma. METHODS: Effects of TTO and terpinen-4-ol on the cellular viability of two tumour cell lines and fibroblast cells were assessed by MTT assay. Induction of apoptotic and necrotic cell death was visualised by fluorescent microscopy and quantified by flow cytometry. Tumour cell ultrastructural changes were examined by transmission electron microscopy and changes in cell cycle distribution were assessed by flow cytometry, with changes in cellular morphology monitored by video time lapse microscopy. RESULTS: TTO and terpinen-4-ol significantly inhibited the growth of two murine tumour cell lines in a dose- and time dependent manner. Interestingly, cytotoxic doses of TTO and terpinen-4-ol were significantly less efficacious against non-tumour fibroblast cells. TTO and terpinen-4-ol induced necrotic cell death coupled with low level apoptotic cell death in both tumour cell lines. This primary necrosis was clarified by video time lapse microscopy and also by transmission electron microscopy which revealed ultrastructural features including cell and organelle swelling following treatment with TTO. In addition, both TTO and terpinen-4-ol induced their inhibitory effect by eliciting G1 cell cycle arrest. CONCLUSION: TTO and terpinen 4-ol had significant anti-proliferative activity against two tumour cell lines. Moreover, the identification of primary necrotic cell death and cell cycle arrest of the aggressive tumour cells highlights the potential anticancer activity of TTO and terpinen-4-ol. PMID- 19680654 TI - Cetuximab-based therapy versus non-cetuximab therapy for advanced cancer: a meta analysis of 17 randomized controlled trials. AB - PURPOSE: To assess the efficacy and safety of cetuximab-based therapy versus non cetuximab therapy for advanced cancer. METHODS: A total of 7,954 patients from 17 randomized controlled trials are identified, with 3,965 patients in the cetuximab group and 3,989 patients in the non-cetuximab group. The outcome was progression free survival (PFS), overall survival (OS), overall response rate (ORR), and grade 3/4 advent events. RESULTS: There was a significant improvement of PFS (HR 0.83, 95%CI 0.78-0.88), OS (HR 0.89, 0.84-0.95), and ORR in the cetuximab group (OR 1.39, 1.22-1.58). In subgroup analysis, in colorectal cancer, there was a significant improvement of PFS (0.72, 0.66-0.78), OS (0.90, 0.81-1.00), and ORR in the cetuximab group (1.36, 1.15-1.60). In head and neck carcinoma, there was a significant improvement of PFS (0.63, 0.54-0.73), OS (0.78, 0.67-0.91), and ORR in the cetuximab group (1.57, 1.15-2.16). In non-small-cell lung cancer, there was a significant improvement of OS (0.86, 0.76-0.96) in the cetuximab group, and no difference on PFS (0.82, 0.64-1.07) and ORR (1.56, 0.85-2.88). In pancreatic cancer, there was no difference on PFS (1.11, 0.97-1.28), OS (1.07, 0.93-1.25), and ORR (0.94, 0.66-1.33). There were higher incidences of grade 3-4 toxicity (OR 1.84), skin-related toxicity (OR 31.80), acneiform rash (OR 30.14), and hypomagnesemia (OR 6.72) in the cetuximab group. CONCLUSIONS: Cetuximab-based therapy improved PFS and OS, and better ORR versus non-cetuximab therapy. The severe adverse events should be predictable and manageable. PMID- 19680655 TI - Limited inter-occasion variability in relation to inter-individual variability in chemotherapy-induced myelosuppression. AB - PURPOSE: A previously developed semi-physiological model of chemotherapy-induced myelosuppression has shown consistent system-related parameter and inter individual variability (IIV) estimates across drugs. A requirement for dose individualization to be useful is relatively low variability between treatment courses (inter-occasion variability [IOV]) in relation to IIV. The objective of this study was to evaluate and compare magnitudes of IOV and IIV in myelosuppression model parameters across six different anti-cancer drug treatments. METHODS: Neutrophil counts from several treatment courses following therapy with docetaxel, paclitaxel, epirubicin-docetaxel, 5-fluorouracil epirubicin-cyclophosphamide, topotecan, and etoposide were included in the analysis. The myelosuppression model was fitted to the data using NONMEM VI. IOV in the model parameters baseline neutrophil counts (ANC0), mean transit time through the non-mitotic maturation chain (mean transit time [MTT]), and the parameter describing the concentration-effect relationship (slope), were evaluated for statistical significance (P < 0.001). RESULTS: Inter-occasion variability in MTT was significant for all the investigated datasets, except for topotecan, and was of similar magnitude (8-16 CV%). IOV in slope was significant for docetaxel, topotecan, and etoposide (19-39 CV%). For all six investigated datasets, the IOV in myelosuppression parameters was lower than the IIV. There was no indication of systematic shifts in the system- or drug sensitivity-related parameters over time across datasets. CONCLUSION: This study indicates that the semi-physiological model of chemotherapy-induced myelosuppression has potential to be used for prediction of the time-course of myelosuppression in future courses and is, thereby, a valuable step towards individually tailored anticancer drug therapy. PMID- 19680656 TI - Epidermal growth factor increases prostaglandin E2 production via ERK1/2 MAPK and NF-kappaB pathway in fibroblast like synoviocytes from patients with rheumatoid arthritis. AB - High concentration of epidermal growth factor (EGF) is found in the synovial fluid of rheumatoid arthritis (RA) that might imply the involvement of EGF in the pathogenesis of arthritic diseases. In order to investigate if EGF is involved in the regulation of cyclooxygenase-2 (COX-2) and the prostaglandin E(2) (PGE(2)) production in fibroblast like synoviocytes (FLS) from patients with RA. The levels of COX-2 and microsomal prostaglandin E synthase-1 (mPGES-1) were evaluated using RT-PCR and Western blot analysis. Electrophoretic mobility shift assay (EMSA) was performed to investigate EGF mediated DNA binding activity of nuclear factor-kappaB (NF-kappaB). PGE(2) levels were analyzed by ELISA. EGF enhanced both COX-2 protein and mRNA expressions. mPGES-1 mRNA level was also increased by EGF treatment. EGF also stimulated ERK1/2 MAPK activity and the inhibition of ERK1/2 by PD098059 (ERK1/2 specific inhibitor) resulted in the suppression of EGF-induced COX-2 expression. The DNA binding activity of NF kappaB was remarkably increased by EGF treatment and the pretreatment of PD098059 abolished EGF-stimulated NF-kappaB activity. We also observed that the level of PGE(2) was significantly elevated with the treatment of EGF in FLS, and the pretreatment of PD098059 abolished this stimulating effect. These results suggest that EGF is involved in the inflammatory process of RA by stimulating COX-2 expression and PGE(2) production. And EGF enhanced PGE(2) production appears to be mediated via ERK1/2 MAPK and NF-kappaB pathway in FLS. PMID- 19680657 TI - Enhancement of tolerance to soft rot disease in the transgenic Chinese cabbage (Brassica rapa L. ssp. pekinensis) inbred line, Kenshin. AB - We developed a transgenic Chinese cabbage (Brassica rapa L. ssp. pekinensis) inbred line, Kenshin, with high tolerance to soft rot disease. Tolerance was conferred by expression of N-acyl-homoserine lactonase (AHL-lactonase) in Chinese cabbage through an efficient Agrobacterium-mediated transformation method. To synthesize and express the AHL-lactonase in Chinese cabbage, the plant was transformed with the aii gene (AHL-lactonase gene from Bacillus sp. GH02) fused to the PinII signal peptide (protease inhibitor II from potato). Five transgenic lines were selected by growth on hygromycin-containing medium (3.7% transformation efficiency). Southern blot analysis showed that the transgene was stably integrated into the genome. Among these five transgenic lines, single copy number integrations were observed in four lines and a double copy number integration was observed in one transgenic line. Northern blot analysis confirmed that pinIISP-aii fusion gene was expressed in all the transgenic lines. Soft rot disease tolerance was evaluated at tissue and seedling stage. Transgenic plants showed a significantly enhanced tolerance (2-3-fold) to soft rot disease compared to wild-type plants. Thus, expression of the fusion gene pinIISP-aii reduces susceptibility to soft rot disease in Chinese cabbage. We conclude that the recombinant AHL-lactonase, encoded by aii, can effectively quench bacterial quorum-sensing and prevent bacterial population density-dependent infections. To the best of our knowledge, the present study is the first to demonstrate the transformation of Chinese cabbage inbred line Kenshin, and the first to describe the effect of the fusion gene pinIISP-aii on enhancement of soft rot disease tolerance. PMID- 19680658 TI - Paraplegia complicating selective steroid injections of the lumbar spine. Report of five cases and review of the literature. AB - BACKGROUND: Selective steroid injections of the lumbar spine carry a risk of paraplegia of sudden onset. Seven cases have been reported in the English literature since 2002. MATERIALS AND METHODS: Five new cases have been analyzed, all coming from Paris area centers. Injections were performed between 2003 and 2008. The following items were searched for: location of a previous lumbar spine surgery if any, symptoms indicating the procedure, route of injection, imaging technique used for needle guidance, injection of a contrast medium, type of steroid, other drugs injected if any, paraplegia level, post-procedure MR findings. The current and reported cases were compared. RESULTS: MR findings were consistent with spinal cord ischemia of arterial origin. The high rate of patients who had been operated on in these cases does not correspond to that of patients undergoing injections. The presence of epidural scar might increase the risk. The foraminal route was the only one involved in nonoperated patients. Foraminal, interlaminar, or juxta-zygoapophyseal routes were used in operated-on patients. CONCLUSION: The high rate of French cases when compared to the literature might arise from the almost exclusive use of prednisolone acetate, a molecule with a high tendency to coalesce in macro-aggregates, putting the spinal cord at risk of arterial supply embolization. PMID- 19680659 TI - Evaluation of the effects of oral water and low-density barium sulphate suspension on bowel appearance on FDG-PET/CT. AB - The purpose of this study is to assess which of five bowel preparation regimes offers superior bowel distension and to assess if these regimes adversely affect FDG activity on PET/CT imaging. The study conformed to HIPAA regulations. Ninety patients were divided into five groups of 18 who received no oral contrast agent (group A); 900 ml of water orally (group B); or 900, 1,350, or 1,800 ml of LDB (groups C, D, E, respectively). PET/CT examinations were assessed quantitatively (bowel diameter, SUV) and qualitatively (visual assessment grading scale) for bowel distension and FDG activity by two blinded readers. ANOVA was utilized to determine if a statistically significant difference (SSD) existed between the groups in terms of distension and FDG uptake. Qualitatively superior bowel distension was observed in group C (LDB) compared to B (water) and greater distension was noted with increased volumes of LDB in C, D, and E. Quantitatively there was an SSD in mean distension between groups C and B (P < 0.001 except duodenum). Qualitatively and quantitatively there was no significant difference in bowel FDG uptake among the groups (P > 0.05). LDB as an oral contrast agent provides superior bowel distension over water and does not induce increased FDG bowel activity. PMID- 19680660 TI - Insertion of double J stent as a therapeutic test in management of adults presenting with loin pain and equivocal ureteropelvic junction obstruction. AB - PURPOSE: To evaluate temporary double J ureteric stenting (TDJS) as a test to diagnose ureteropelvic junction obstruction (UPJO) in equivocal cases. MATERIALS AND METHODS: One hundred and nine consecutive adult patients with loin pain and ipsilateral equivocal UPJO (EqUPJO) on radioisotope diuretic renogram (RDR) were enrolled in the present study. All patients underwent TDJS for 3 weeks. According to the response (relief of pain) to TDJS, which was assessed by visual analogue pain score (VAPS), patient preference and further management, patients were classified into five groups: Group Ia non-responders who elected conservative treatment for their condition, Group Ib non-responders who elected intervention, Group IIa responders who elected conservative treatment and Group IIb responders who elected intervention. Last group patients were randomized to either early (Group IIb(1)) or delayed intervention 3 months later (Group IIb(2)). Intervention was in the form of pyeloplasty (24 patients) and endopyelotomy (38 patients). All patients were followed up by measuring VAPS and RDR. RESULTS: After at least 6 months from management decision, 97 patients were available for evaluation. The VAPS dropped by 21.25% (P < 0.001), 32% (P = 0.004), 2% (P = 0.6), 54% (P < 0.001) and 65% (P < 0.001) in groups Ia, Ib, IIa, IIb1, and IIb2, respectively. On the other hand the T (1/2) of RDR dropped significantly only in groups Ib, IIb1, and IIb2 by a mean of 6.5 min (P = 0.005), 8.02 min (P < 0.001), and 7.3 min (P < 0.001), respectively. CONCLUSION: TDJS in cases of EqUPJO with loin pain is helpful in defining cases suitable for intervention versus conservative treatment. PMID- 19680661 TI - Quantitative evaluation of electrodes for external urethral sphincter electromyography during bladder-to-urethral guarding reflex. AB - PURPOSE: Accuracy in the recording of external urethral sphincter (EUS) electromyography (EMG) is an important goal in the quantitative evaluation of urethral function. The aim of this study was to quantitatively compare electrode recordings taken during tonic activity and leak point pressure (LPP) testing. METHODS: Several electrodes, including the surface electrode (SE), concentric electrode (CE), and wire electrode (WE), were placed on the EUS singly and simultaneously in six female Sprague-Dawley rats under urethane anesthesia. The bladder was filled via a retropubic catheter while LPP testing and EUS EMG recording were done. Quantitative baseline correction of the EUS EMG signal was performed to reduce baseline variation. Amplitude and frequency of 1-s samples of the EUS EMG signal were measured before LPP (tonic activity) and during peak LPP activity. RESULTS: The SE, CE, and WE signals demonstrated tonic activity before LPP and an increase in activity during LPP, suggesting that the electrodes accurately recorded EUS activity during tonic activity and during the bladder-to EUS guarding reflex, regardless of the size or location of detection areas. SE recordings required significantly less baseline correction than both CE and WE recordings. The activity in CE-recorded EMG was significantly higher than that of the SE and WE both in single and simultaneous recordings. CONCLUSIONS: These electrodes may be suitable for testing EUS EMG activity. The SE signal had significantly less baseline variation and the CE detected local activity more sensitively than the other electrodes, which may provide insight into choosing an appropriate electrode for EUS EMG recording. PMID- 19680662 TI - [Data security and the handling of patient data in home monitoring systems]. AB - Data security must be considered seriously in the context of telemedical home monitoring because of the transmission and communication of patients' personal data. The contract governing medical treatment allows the ophthalmologist to process all data relevant to treatment. In Germany the legal framework for this purpose is provided by the Data Protection Act, various German hospital acts, and codes of medical professional conduct. In principle, these rules apply to telemedical home monitoring as well as to common physician-patient relationships. The patient must be informed extensively in an understandable manner and must give his or her written consent. However, the advanced options of new IT technologies demand the development of technical and organizational concepts that guarantee compliance with legal and regulatory affairs, assure data security, and prevent data abuse. PMID- 19680663 TI - Cytogenetic evaluation of neuroblastoma using fine needle aspiration cultures. AB - PURPOSE: Open surgical biopsies of the primary tumor and bone marrow sampling in metastatic disease are the major sources of tumor material for genetic studies in neuroblastoma. The possibility of using cultures of material procured through Fine Needle Aspiration (FNA) for upfront cytogenetic studies of neuroblastomas is studied. METHODS: From January 2006 to April 2008, 18 patients were diagnosed as neuroblastoma on fine needle aspiration cytology (FNAC). FNA material was cultured and processed to obtain metaphases and chromosomal analysis was performed. Minimum of five selected metaphases were analyzed and changes were documented. RESULTS: The diagnosis of neuroblastoma was confirmed by FNAC in all. Among the 18 cases, 9 cases (50%) showed metaphases on cultures. Four cultures did not grow and five cultures got contaminated. Out of the nine cases that showed metaphases, four cases showed cytogenetic abnormalities. Near triploidy and Double minutes (Dmins) were seen in one case and tetraploidy in another case. 1q gain and 2q gain were observed respectively in third and fourth cases. CONCLUSION: FNA provides viable single cell suspension suitable for cultures. This may be a good alternative strategy to acquire material for cytogenetic studies in neuroblastoma. PMID- 19680664 TI - Hypoganglionic colorectum in the chick embryo: a model of human hypoganglionosis. AB - INTRODUCTION: The enteric nervous system is an intrinsic network of nerve cells and glia within the gastrointestinal wall, which originates in the vagal and sacral neural tube. The vagal neural tube is known to supply the colorectum with the majority of its nerve cells, and its ablation during early development produces a hypoganglionic colorectum. We hypothesized that the cholinergic nerve activity in the chick embryo hypoganglionic colorectum is decreased similar to the human situation and, therefore, this study is designed to investigate cholinergic innervations in the chick embryo hypoganglionic colorectum. MATERIALS AND METHODS: Chicken eggs were incubated until embryos reached the 10-12 somite stage. The vagal neural tube was microsurgically ablated and eggs were returned to the incubator until embryos reached E12 and E14. Whole embryos were fixed and embedded in paraffin wax. Transverse sections were cut and immunohistochemistry was performed using a neural crest cell antibody, human natural killer-1 (HNK-1), and a choline acetyltransferase antibody (ChAT). RESULTS: The results showed that in normal embryos, the colorectum contained many nerve cells (HNK-1) and ChAT positive nerve cells and fibres, while in embryos with a hypoganglionic colorectum, the number of nerve cells (HNK-1) and ChAT-positive nerve cells and fibres was decreased. CONCLUSION: Cholinergic nerve activity is decreased as a result of a reduction in nerve cell numbers in the chick embryo colorectum. These results suggest that the cholinergic activity in the hypoganglionic chick model resembles that of human hypoganglionosis. PMID- 19680665 TI - Long-term outcome of internal sphincter myectomy in patients with internal anal sphincter achalasia. AB - BACKGROUND: Internal anal sphincter achalasia (IASA) is a condition with presentation similar to Hirschsprung's disease (HD), but with the presence of ganglion cells on rectal suction biopsy (RSB). The diagnosis is made on anorectal manometry (ARM) by the absence of the rectosphincteric reflex on rectal balloon inflation. Internal sphincter myectomy (ISM) is the treatment of choice for patients with IASA. Recently, botulinum toxin has been used to treat IASA patients. The purpose of this study was to assess the long-term bowel function in patients with IASA following ISM. METHODS: The medical records of 24 patients with IASA managed by ISM during 1993-2005 were examined. There were 18 boys and 6 girls, aged 2-12 years. All patients presented with intractable constipation with or without soiling. The diagnosis was made by the demonstration of the absence of the rectosphincteric reflex on ARM. HD was excluded by the presence of ganglion cells and normal acetylcholinesterase activity in RSB. Patients were followed 4 14 years later. RESULTS: Fifteen (62.5%) patients at the time of follow-up had regular bowel motions without the use of laxatives. Six (25%) patients had regular bowel motions, but remained on small doses of laxatives. Two (8.3%) patients who suffered from constipation and soiling required twice weekly enemas to remain clean. One (4.2%) patient required resection of dilated rectosigmoid colon 3 years after myectomy, remains on laxatives, but has normal bowel control. No patients had faecal incontinence following ISM. CONCLUSION: This long-term follow-up study shows that the vast majority of IASA patients have normal bowel control following ISM. PMID- 19680666 TI - Tethered cord in patients with anorectal malformation: preliminary results. AB - BACKGROUND: A tethered cord (TC) has been reported in as much as 50% of the patients affected by anorectal malformation (ARM). No guidelines for timing and modality of diagnosis and treatment have been established. We present the preliminary results of a multidisciplinary protocol carried out at our center. METHODS: Seventy-four ARM patients underwent spinal magnetic resonance imaging (MRI). All TC patients underwent videourodynamic (UD), somatosensory-evoked potentials (SEPs), and neurological examination at baseline and, if normal, at 5 and 10 years of age. Conversely, when UD or SEP abnormalities were detected the follow-up was individually tailored at shorter time. RESULTS: 25/74 patients had a neuroradiological TC (33.7%). Based on the results of UD, SEP, and neurological status, four patients were untethered, eight are possible candidates, nine are stable, and four were excluded because of incomplete data. DISCUSSION: Tethered cord is frequent in ARM patients. Because neurological deficits secondary to TC can contribute to neurological disability, we recommend routine MRI examination and a multidisciplinary program of follow-up in cases of TC. Preliminary results suggest the combined use of SEPs and UD could represent a useful adjunct to clinical examination in patients in whom a "wait and see" approach is preferred to the prophylactic surgery. PMID- 19680667 TI - Prospective long-term functional and cosmetic results of ASARP versus PASRP in treatment of intermediate anorectal malformations in girls. AB - PURPOSE: Prospective randomized comparison of the functional and cosmetic results of anterior sagittal anorectoplasty (ASARP) and posterior sagittal anorectoplasty (PSARP) in the management of intermediate anorectal malformations (ARMs) in girls was performed. METHODS: Thirty-eight girls with intermediate ARMs were randomly allocated to group A treated by ASARP technique and group B treated by PSARP technique: 14 girls from each group passed 30-month age for early functional assessment by Templeton and Holschneider scores, 18 cases passed 55 months for late functional evaluation. Results were compared statistically where P value < or = 0.05 was considered significant. RESULTS: Cosmetic satisfaction was higher in ASARP group with a median score of 3 compared to 2 in PSARP group. Functional results were assessed early at median of 33.5 months post-repair where Templeton score was 4 versus 3.75 and Holschneider score was 11 versus 11. Late functional assessment was done for 8 cases from each group where Templeton score was 4 versus 4, while Holschneider score was 12 versus 11. CONCLUSIONS: ASARP is an optimal technique for treatment of intermediate ARM in girls. Cosmetic results were superior to PSARP. Despite the observations that functional results give higher scores in ASARP group versus PSARP group, no statistical significance could be reached. PMID- 19680668 TI - The effect of peri-stomal infiltration with bupivacaine/epinephrine on post operative pain, nausea and ease of surgery in reversal of loop ileostomies. AB - PURPOSE: Evidence to support the routine use of local anaesthetic in the reversal of loop ileostomy is equivocal. This randomized control study looked at the use of peri-operative infiltration of stoma with 0.25% bupivacaine with 1/200,000 epinephrine on the ease of surgery and its effect on post-operative pain and nausea. METHODS: Sixty patients were randomized to receive peri-stomal infiltration with either 0.25% bupivacaine with 1/200,000 epinephrine or normal saline. The surgeon graded the surgery as straightforward, intermediate or difficult, and the time for the operation was also recorded. Post-operatively, analgesia was provided via PCA for 24 h. Post-operative pain and nausea scores and total morphine usage median (inter-quartile range) were compared using the Mann-Whitney U test with p < 0.05 considered significant. RESULTS: There was no difference between the local anaesthetic groups and controls with respect to opiate consumption (p = 0.4), post-operative pain (p = 0.72) or nausea (p = 0.78). Shorter total anaesthetic and operative times were noted in study group, but this was not significant (p = 0.55). However, surgery was found to be easier in the local anaesthetic group (p = 0.0046). CONCLUSION: Peri-stomal infiltration with 0.25% bupivacaine with 1/200,000 epinephrine does not impact on post operative pain and nausea scores or opiate analgesia use. However, its use is recommended as an aid to dissection in surgery. PMID- 19680669 TI - [ASAS classification criteria for axial spondyloarthritis]. PMID- 19680670 TI - [Physiotherapy and exercise in osteoporosis and its complications]. AB - Osteoporosis is defined by decreased bone strength and increased susceptibility to fracture. Fractures and their consequences are the clinical manifestation of osteoporosis. Acute and chronic pain, functional limitations including permanent impairment and the need for long-term care may be caused by osteoporotic fractures. The aim of osteoporosis treatment is to prevent fractures by bone strengthening. The aims of rehabilitation in patients with osteoporosis are to reduce pain, maximize the level of musculoskeletal function, particularly following fractures, decrease risk of falls and optimize quality of life and independence. Certain sports and exercises greatly promote skeletal development in children and adolescents and augment bone strength in adults. Physiotherapy and therapeutic exercise may relieve pain, increase musculoskeletal function and form an important part of fall management. PMID- 19680671 TI - Evidence for antibody-mediated pathogenesis in anti-NMDAR encephalitis associated with ovarian teratoma. AB - We report the immunopathological analysis of the brain and tumor of two patients who died of anti-NMDAR-associated encephalitis, and of the tumor of nine patients who recovered. Findings included prominent microgliosis and deposits of IgG with rare inflammatory infiltrates in the hippocampus, forebrain, basal ganglia, and spinal cord. Detection of cells expressing markers of cytotoxicity (TIA, granzyme B, perforin and Fas/Fas ligand) was extremely uncommon. All tumors showed NMDAR expressing neurons and inflammatory infiltrates. All patients' NMDAR antibodies were IgG1, IgG2, or IgG3. No complement deposits were observed in any of the central nervous system regions examined. Overall, these findings coupled with recently reported in vitro data showing that antibodies downregulate the levels of NMDA receptors suggest that the antibody immune-response is more relevant than cytotoxic T-cell mechanisms in the pathogenesis of anti-NMDAR-associated encephalitis. PMID- 19680672 TI - Microcirculatory sequelae of the rotator cuff after antegrade nailing in proximal humerus fracture. AB - INTRODUCTION: Antegrade nailing allows a stable fixation and, thus, an early functional after treatment in proximal humerus fractures. Since the surgical procedure in antegrade humeral nailing requires a split of the supraspinatus tendon, the question arises whether the surgical approach causes microcirculatory dysfunction of the tendon. MATERIALS AND METHODS: A total of 15 consecutive patients suffering from proximal humerus fractures were enrolled. During the implantation of an antegrade humerus nail, microvascular perfusion of the supraspinatus tendon was directly visualized after the exposition and stabilization of the fracture using the OPS-imaging technique. RESULTS: Immediately after exposure, the nutritive perfusion showed physiological values of tendon microcirculation. After implanting antegrade humeral nails, the perfusion of the supraspinatus tendon reduced markedly. Capillary width was unaffected by the surgical procedure. CONCLUSION: The trauma leading to proximal humerus fracture causes no fundamental impairment of nutritive perfusion of the rotator cuff. Whereas the implantation of an antegrade humerus nail, which necessarily includes a splitting of the rotator cuff, nearly halves the functional capillary density of the supraspinatus tendon. Even though this effect seems to be reversible, the surgical dissection of the supraspinatus tendon should be performed in a soft tissue sparing way. PMID- 19680673 TI - Severe menopausal symptoms in middle-aged women are associated to female and male factors. AB - BACKGROUND: The frequency and intensity of menopausal symptoms within a given population, as assessed by several tools, may vary and depend on several factors, such as age, menopausal status, chronic conditions and personal and partner socio demographic profile. OBJECTIVE: To determine the frequency and intensity of menopausal symptoms and related risk factors among middle-aged women. METHODS: In this cross-sectional study a total of 404 women aged 40 to 59 years, visiting inpatients at the Enrique C. Sotomayor Gynecology and Obstetrics Hospital, Guayaquil, Ecuador, were requested to fill out the menopause rating scale (MRS) and a questionnaire containing personal and partner data. RESULTS: Mean age of surveyed women (n=404) was 48.2+/-5.7 years, 85.1% had 12 or less years of schooling and 44.8% were postmenopausal. None was on hormonal therapy (HT) for the menopause or psychotropic drugs. Regarding their partner, erectile dysfunction was present in 23.8%, premature ejaculation in 21.2% and 43.5% abused alcohol. The four most frequently found symptoms of those composing the MRS were muscle and joint problems (80%), depressive mood (73.5%), physical and mental exhaustion (71.3%) and irritability (68%). Mean total MRS score was 18+/-10.6 (median 17) and for subscales: 7.2+/-4.5 (somatic); 6.9+/-4.8 (psychological) and 3.9+/-3.4 (urogenital). Women presented severe scores in 53, 36.1, 48.3 and 49.8% for total MRS and somatic, psychological and urogenital subscales, respectively. After adjusting for confounding factors, logistic regression analysis determined that female higher parity and partner premature ejaculation increased the risk for presenting severe total MRS scores (impaired female quality of life), whereas women who had a positive perception of their health status were at decreased risk. CONCLUSION: In this middle-aged series psychological menopausal symptoms were the most frequent in which severity was associated to parity and partner sexual dysfunction. PMID- 19680674 TI - Evaluation of external exposures of the population of Ozyorsk, Russia, with luminescence measurements of bricks. AB - Recently discovered historical documents indicate that large releases of noble gases (mainly (41)Ar and radioactive isotopes of Kr and Xe) from the Mayak Production Association (MPA) over the period from 1948 to 1956 may have caused considerable external exposures of both, inhabitants of Ozyorsk and former inhabitants of villages at the upper Techa River. To quantify this exposure, seven brick samples from three buildings in Ozyorsk, located 8-10 km north northwest from the radioactive gas release points, were taken. The absorbed dose in brick was measured in a depth interval of 3-13 mm below the exposed surface of the bricks by means of the thermoluminescence (TL) and the optically stimulated luminescence (OSL) method. Generally, luminescence properties using TL were more favorable for precise dose determination than using OSL, but within their uncertainties the results from both methods agree well with each other. The absorbed dose due to natural radiation was assessed and subtracted under the assumption of the bricks to be completely dry. The weighted average of the anthropogenic dose for all samples measured by TL and OSL is 10 +/- 9 and 1 +/- 9 mGy, respectively. An upper limit for a possible anthropogenic dose in brick that would not be detected due to the measurement uncertainties is estimated at 24 mGy. This corresponds to an effective dose of about 21 mSv. A similar range of values is obtained in recently published dispersion calculations that were based on reconstructed MPA releases. It is concluded that the release of radioactive noble gases from the radiochemical and reactor plants at Mayak PA did not lead to a significant external exposure of the population of Ozyorsk. In addition, the study demonstrates the detection limit for anthropogenic doses in ca. 60-year-old bricks to be about 24 mGy, if luminescence methods are used. PMID- 19680675 TI - Amerindian mitochondrial DNA haplogroups predominate in the population of Argentina: towards a first nationwide forensic mitochondrial DNA sequence database. AB - The study presents South American mitochondrial DNA (mtDNA) data from selected north (N = 98), central (N = 193) and south (N = 47) Argentinean populations. Sequence analysis of the complete mtDNA control region (CR, 16024-576) resulted in 288 unique haplotypes ignoring C-insertions around positions 16193, 309, and 573; the additional analysis of coding region single nucleotide polymorphisms enabled a fine classification of the described lineages. The Amerindian haplogroups were most frequent in the north and south representing more than 60% of the sequences. A slightly different situation was observed in central Argentina where the Amerindian haplogroups represented less than 50%, and the European contribution was more relevant. Particular clades of the Amerindian subhaplogroups turned out to be nearly region-specific. A minor contribution of African lineages was observed throughout the country. This comprehensive admixture of worldwide mtDNA lineages and the regional specificity of certain clades in the Argentinean population underscore the necessity of carefully selecting regional samples in order to develop a nationwide mtDNA database for forensic and anthropological purposes. The mtDNA sequencing and analysis were performed under EMPOP guidelines in order to attain high quality for the mtDNA database. PMID- 19680676 TI - Avastin use in high risk corneal transplantation. AB - BACKGROUND: Corneal neovascularization is a major risk factor for graft failure after corneal transplantation. The purpose of this study was to investigate the effect of subconjunctival, perilimbal, and/or intrastromal bevacizumab (Avastin) on corneal neovascularization in patients with penetrating keratoplasty (PKP). METHODS: Fourteen eyes of 14 patients with high risk corneal transplantation and corneal neovascularization after PKP (nine men and five women with a mean age of 63 years) were included in this non-comparative interventional case series. Indications for PKP were: vascularized leukomas after herpetic keratitis and chemical burn, advanced pseudophakic bullous keratopathy with superficial and deep corneal vascularization, keratoconus, severe infection in hereditary corneal dystrophy, and failed corneal grafts. Subconjunctival, perilimbal, and/or intrastromal bevacizumab of dose of 2.5 mg/ 0.1 ml/ per affected quadrant was injected at the site of neovascularization in each patient at the end of surgery and/or at follow up visits. One or two injections were applied. At each visit a full eye examination with photo documentation was performed. Follow-up period was 2 to 8 months (mean 7.1 months). RESULTS: Decrease of corneal neovascularization was observed in eleven patients (78.6%). Regression of neovascularization with fading of small vessels was demonstrated. Despite high- risk patient pool, twelve grafts (85.7%) remained transparent for the period of observation, and patients maintained good visual acuity. In two patients with initial graft rejection and vascularization, subconjunctival and perilimbal application of bevacizumab was beneficial in overcoming the corneal inflammation and initial rejection. No adverse reactions have been detected to date in patients with subconjunctival, perilimbal, and/or intrastromal injection of bevacizumab. CONCLUSIONS: Bevacizumab is an efficient and safe additional treatment option for improvement of prognosis in high-risk corneal transplantation with pre- and postoperative corneal neovascularization. PMID- 19680677 TI - Antifibrotic effects of tocotrienols on human Tenon's fibroblasts. AB - PURPOSE: To compare the antifibrotic effect of vitamin E isoforms alpha-, gamma-, and delta-tocotrienol on human Tenon's fibroblasts (hTf) to the antimetabolite mitomycin C. METHODS: Antifibrotic effects of alpha- (40, 60, 80, 100, and 120 microM), gamma- (10, 20, 30, and 40 microM) and delta-tocotrienol (10, 20, 30, and 40 microM) on hTf cultures were evaluated by performing proliferation, migration and collagen synthesis assays. Whereas for vitamin E the exposure time was set to 7 days to mimic subconjunctival application, cultures were exposed only 5 min to mitomycin C 100 microg/ml to mimic intraoperative administration. Cell morphology (phase contrast microscopy) as an assessment for cytotoxicity and cell density by measuring DNA content in a fluorometric assay to determine proliferation inhibition was performed on day 0, 4, and 7. Migration ability and collagen synthesis of fibroblasts were measured. RESULTS: All tested tocotrienol isoforms were able to significantly inhibit hTf proliferation in a dose-dependent manner (maximal inhibitory effect without relevant morphological changes at day 4 for alpha-tocotrienol 80 microM with 36.7% and at day 7 for alpha-tocotrienol 80 microM with 42.6% compared to control). Degenerative cell changes were observed in cultures with concentrations above 80 microM for alpha- and above 30 microM for gamma- and delta-tocotrienol. The highest collagen synthesis inhibition has been found with 80 microM alpha-tocotrienol (62.4%) and no significant inhibition for mitomycin C (2.5%). Migration ability was significantly reduced in cultures exposed to 80 microM alpha- and 30 microM gamma-tocotrienol (inhibition of 82.2% and 79.5%, respectively, compared to control) and also after mitomycin C treatment (60.0%). Complete growth inhibition without significant degenerative cell changes could only be achieved with mitomycin C. CONCLUSION: In vitro, all tested tocotrienol isoforms were able to inhibit proliferation, migration and collagen synthesis of human Tenon's fibroblasts and therefore may have the potential as an anti-scarring agent in filtrating glaucoma surgery. PMID- 19680678 TI - Effects of folic acid and homocysteine on spinal cord morphology of the chicken embryo. AB - Maternal ingestion of folic acid (FA) reduces neural tube defects, which are associated with high homocysteine levels. Present study evaluated the effects of FA and homocysteine on cell proliferation and cell adhesion, as well as on apoptosis, throughout the development of the spinal cord and mesenchyme of chicken embryos. Normal closure of the neural tube and a regular distribution of the mesenchymal cells were observed in control and FA-treated embryos. All homocysteine-treated embryos and also 6 of 10 embryos treated with FA+homocysteine showed failure of closure of the neural tube. Homocysteine decreased the thickness of the mantle and marginal layers of the spinal cord, and FA did not prevent this effect. FA treatment reversed the decrease of proliferating cells in the spinal cord induced by homocysteine. FA-treated embryos showed the highest numerical density of apoptotic cells. Homocysteine treatment reduced NCAM expression in both spinal cord and mesenchymal tissue, and FA prevents this effect. These results are important because they demonstrate in situ that the imbalance between FA and homocysteine levels can lead to disruptions in spinal cord development, changing proliferation, apoptosis, and cell adhesion and consequently changing the arrangement of the spinal cord layers. PMID- 19680680 TI - Effect of exercise duration on session RPE at an individualized constant workload. AB - Session RPE (SRPE) permits subjective ratings of global effort following exercise. SRPE is useful for evaluating training load; however, mediating factors are not well understood. This study compared SRPE among treadmill trials at a clamped workload with a varied duration. Ten subjects (VO(2)max: 48.9 +/- 10.5 ml kg(-1) min(-1)) completed a maximal-exertion treadmill trial followed by treadmill trials of 20 (TM20), 30-(TM30), and 40-(TM40) min (counterbalanced) duration at approximately 70% individualized VO(2)max. Heart rate (HR) and acute RPE (during exercise) were recorded every 5 min with blood lactate concentration [La] recorded before initiating exercise and every 10 min throughout exercise to completion. Analyses showed no significant differences among TM20, TM30, and TM40 for mean HR, [La] or acute RPE which isolated effects of duration on SRPE. Session RPE (recorded 20 min post exercise) relative to overall (SRPEO), legs (SRPEL) and breathing (SRPEB) was analyzed using a 3 (trial) x 3 (SRPE: O, L, B) repeated measures ANOVA (alpha 0.05). There was no main effect for duration, but there was a significant main effect for SRPE type with SRPEO (4.40 +/- 2.18) greater than SRPEB (3.56 +/- 1.98) but no difference for SRPEL (4.43 +/- 2.16). Follow-up tests for within-trial differences showed no significant differences for SRPEL versus SRPEO, but blunted SRPEB estimations (vs. SRPEO and SRPEL). Primary results indicate affects of duration on SRPE are minimal. Further, within the limits of the current study, differentiating SRPE estimations do not appear advantageous; however, further work is warranted. PMID- 19680679 TI - Climbing time to exhaustion is a determinant of climbing performance in high level sport climbers. AB - We studied which physiological and kinanthropometric characteristics determine climbing performance in 16 high-level sports climbers aged 29.9 +/- 4.9 years. Body composition parameters were measured with dual energy X-ray absorptiometry scanner. We also measured kinanthropometric and physical fitness parameters. The sex-specific 75th percentile value of onsight climbing ability was used to divide the sample into expert (<75th) and elite (> or =75th) climbers. All the analyses were adjusted by sex. The 75th percentile value of onsight climbing ability was 7b in women and 8b in men. There were no differences between expert and elite climbers in the studied variables, except in climbing time to exhaustion and bone mineral density. Elite climbers had a significantly higher time to exhaustion than the expert group (770.2 +/- 385 vs. 407.7 +/- 150 s, respectively, P = 0.001). These results suggest that, among climbers with a high level of performance, as those analysed in this study, climbing time to exhaustion is a major determinant of climbing performance. PMID- 19680681 TI - Isometric handgrip exercise improves acute neurocardiac regulation. AB - Isometric handgrip (IHG) training (>6 weeks) has been shown to reduce resting arterial blood pressure (ABP) and improve cardiac autonomic modulation. However, the effects of a single bout of IHG on acute neurocardiac regulation remain unknown. The purpose of this study was to examine the effect of IHG exercise on nonlinear heart rate dynamics and cardiac vagal activity. Nonlinear dynamics were assessed by sample entropy, detrended fluctuation analysis (alpha(1)), and correlation dimension techniques. The 4-second exercise test was used to calculate the cardiac vagal index (CVI), an indirect measure of cardiac vagal activity. In a randomized crossover design, 18 older (70 +/- 5 years of age) subjects completed IHG exercise (four 2-min isometric contractions at 30% MVC) and a time-matched control condition. Following a single bout of bilateral IHG, there was a small reduction in systolic blood pressure (125 +/- 2 to 122 +/- 1 mmHg, P < 0.01), in addition to, a significant decrease in alpha(1) (1.42 +/- 0.12 to 1.22 +/- 0.10, P < 0.05), an increase in sample entropy (1.28 +/- 0.03 to 1.40 +/- 0.05, P < 0.001), and an increase in the CVI (1.24 +/- 0.03 to 1.29 +/- 0.03, P < 0.01). These results suggest improvements in acute cardiac autonomic modulation following a single bout of IHG. This may be mechanistically linked to the observed reductions in ABP seen in previous IHG training studies. Alternatively, these acute effects may have clinical applications and require further investigation. PMID- 19680682 TI - Differential atrophy of the lower-limb musculature during prolonged bed-rest. AB - Patients with medical, orthopaedic and surgical conditions are often assigned to bed-rest and/or immobilised in orthopaedic devices. Although such conditions lead to muscle atrophy, no studies have yet considered differential atrophy of the lower-limb musculature during inactivity to enable the development of rehabilitative exercise programmes. Bed-rest is a model used to simulate the effects of spaceflight and physical inactivity. Ten male subjects underwent 56 days of bed-rest. Magnetic resonance imaging of the lower-limbs was performed at 2-weekly intervals during bed-rest. Volume of individual muscles of the lower limb and subsequently, rates of atrophy were calculated. Rates of atrophy differed (F = 7.4, p < 0.0001) between the muscles with the greatest rates of atrophy seen in the medial gastrocnemius, soleus and vastii (p < 0.00000002). The hamstring muscles were also affected (p < 0.00015). Atrophy was less in the ankle dorsiflexors and anteromedial hip muscles (p > 0.081). Differential rates of atrophy were seen in synergistic muscles (e.g. adductor magnus > adductor longus, p = 0.009; medial gastrocnemius > lateral gastrocnemius, p = 0.002; vastii > rectus femoris, p = 0.0002). These results demonstrate that muscle imbalances can occur after extended periods of reduced postural muscle activity, potentially hampering recovery on return to full upright body position. Such deconditioned patients should be prescribed "closed-chain" simulated resistance exercises, which target the lower-limb antigravity extensor muscles which were most affected in bed-rest. PMID- 19680683 TI - Lysophosphatidylcholine- and MCP-1-induced chemotaxis of monocytes requires potassium channel activity. AB - One of the earliest cellular responses in atherogenesis is the focal recruitment of circulating monocytes, while the most important atherogenic chemoattractants are monocyte chemoattractant protein-1 (MCP-1) and lysophosphatidylcholine (LPC). Invading monocytes transform into activated macrophages and foam cells, which stimulate inflammatory processes and promote atherosclerosis. In this study, we have searched for common mechanisms involved in MCP-1- and LPC-stimulated monocyte migration. We have found that migration of THP-1 monocytes stimulated with MCP-1 was reduced upon inhibition of G(i/o) proteins with pertussis toxin and upon inhibition of platelet activating factor receptors with BN52021, whereas LPC-stimulated monocyte chemotaxis remained unaffected by both inhibitors. Furthermore, Cl(-) channels were only required for MCP-1-induced chemotaxis. However, activity of voltage-gated K+ channels and of Ca2+-activated K+ channels was found to be involved in migration of monocytes stimulated with either MCP-1 or LPC. Inhibition of voltage-gated K+ channels with 4-aminopyridine or margatoxin partially inhibited MCP-1- and LPC-stimulated migration of monocytes. Blockade of Ca2+-activated K+ channels with TRAM-34 also partially reduced migration of MCP-1- and LPC-stimulated monocytes. Simultaneous inhibition of voltage-gated and Ca2+-activated K+ channels abolished MCP-1- and LPC-induced chemotaxis of monocytes. Thus, K+ channel inhibition may represent a novel powerful strategy to reduce monocyte infiltration and subsequent inflammation in atherosclerosis. PMID- 19680684 TI - A technique for monitoring multiple signals with a combination of prism-based total internal reflection fluorescence microscopy and epifluorescence microscopy. AB - Physiological phenomena are regulated by multiple signal pathways upon receptor stimulation. Here, we have introduced a new technique with a combination of prism based total internal reflection fluorescence microscopy (PBTIRFM) and epifluorescence microscopy (EPI) to simultaneously monitor multiple signal pathways. This instrumentation allows us to visualize three signal pathways, Ca2+, cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA), and diacylglycerol (DAG)/protein kinase C (PKC) signals in living cells. Three fluorescent indicators were employed for this purpose: (1) Fura-2 AM as a calcium sensor; (2) Epac1-camp, a cyan fluorescent protein-yellow fluorescent protein fluorescence resonance energy transfer-based cAMP indicator, as a cAMP sensor; and (3) C1-tagged monomeric red fluorescent protein, a tandem DAG-binding domain of PKC gamma, as a DAG sensor or myristoylated alanine-rich C kinase substrate tagged DsRed for the PKC activation pathway. The DAG signal was monitored by PBTIRFM, whereas the Ca2+ and cAMP signals were monitored by EPI. Adenosine trisphosphate resulted in generation of all three second messengers in triple probe-loaded Cos-7 cells. The spectral overlap between these signal probes was evaluated by means of linear unmixing. Forskolin also evoked Ca2+, cAMP/PKA, and DAG/PKC signals whereas acetylcholine activated Ca2+ and DAG/PKC signals as well as inhibiting cAMP generation in triple probe-loaded insulin-secreting cells. Thus, the optical observation system combining PBTIRFM and EPI offers a great advance in analyzing interplay of multiple signaling pathways, such as these second messengers, upon G-protein-coupled receptor stimulation in living cells. PMID- 19680685 TI - Immunohistochemical expression of ubiquitin and telomerase in cervical cancer. AB - Ubiquitin and telomerase immunohistochemical expression patterns in cervical cancer were compared with normal cervical tissue samples. Eighty-one cervical cancer cases and 22 normal exo-endocervical tissue were examined with polyclonal antibody for ubiquitin and 44G12 clone for telomerase using tissue microarrays. The results were interpreted using a semiquantitative scale The average age of patients was 50.67 years. The most frequent histological types were moderately differentiated epidermoid carcinoma (43.5%), according to the degree of differentiation, and endocervical adenocarcinoma (42.1%). Immunohistochemical findings were as follows: 98.7% of cervical cancers showed immunoexpression for ubiquitin and 52.6% for telomerase. Statistically significant differences were found in tumor immunoreactivity when compared with control tissue (p < 0.0007) for both biomarkers. There was no significant difference in biomarker expression at different histological types of tumors, although telomerase was less expressed in endocervical adenocarcinoma. Our findings confirm that abnormal immunoexpression pattern of ubiquitin and telomerase is common in HPV-positive cervical cancer, indicating the existence of an intense degradation of proteins, subsequent cellular immortalization and maintenance of the malignant phenotype. PMID- 19680687 TI - Biochemical efficacy of N-carbamylglutamate in neonatal severe hyperammonaemia due to propionic acidaemia. PMID- 19680686 TI - Adhesion molecules and p16 expression in endocervical adenocarcinoma. AB - An immunohistochemical (IHC) study has been conducted on 34 cases of untreated endocervical adenocarcinomas collected among three institutions (Ospedale S. Andrea, Rome; Istituto Nazionale Tumori "Fondazione G. Pascale", Naples; and Clinica Malzoni, Avellino). The E-cadherin and alpha- and beta-catenin complex status has been investigated along with p16INK4a in all studied cases with the aim to study whether the pattern of expression of the cadherin-catenin complex could be causally related to the expression of P16INK4a protein. Results were evaluated for statistical significance by a non-parametric test (Kruskal-Wallis). Endocervical adenocarcinomas as a group were uniformly expressing p16INK4a except for two cases, and all lesions displayed downregulation of the cadherin-catenin complex, without demonstrating statistically significant differences among the different histotypes. The lack of nuclear accumulation of beta-catenin found in this group of lesions probably implies that no alteration of the beta-catenin/Wnt metabolic pathway is present in endocervical adenocarcinoma, as opposed to what is found in the literature for squamous carcinoma of the cervix. The diffuse expression of p16INK4a protein in this group of neoplasms stresses the important role of high-risk human papillomavirus infection in neoplastic causation possibly via the viral E7-mediated inactivation of pRB tumor-suppressor protein and also underlines the useful role of p16INK4a immunostaining in the diagnostic algorithm of endocervical adenocarcinomas. In consideration of these findings, investigation of downstream beta-catenin genes c-myc and cyclin D1 is sought as possibly contributive in the molecular pathogenesis of endocervical adenocarcinoma. PMID- 19680688 TI - Posttreatment surveillance after paraaortic radiotherapy for stage I seminoma: a systematic analysis. AB - CONTEXT: The extent and duration of routine follow-up after paraaortic (PA) radiotherapy for stage I seminoma remain controversial in terms of efficacy, costs of technical investigations and long-term morbidity. OBJECTIVE: To analyze the current literature assessing routine follow-up after PA radiotherapy for stage I seminoma. EVIDENCE ACQUISITION: We identified all published reports on PA radiotherapy for stage I seminoma (1986-2005). We analyzed time patterns of recurrence, sites and methods of detection of relapse, and follow-up programs used. EVIDENCE SYNTHESIS: We identified 11 publications reporting outcome in 2,280 patients. Median time to recurrence in 80 relapsing patients was 15.5 months. Less than 10% of recurrences were diagnosed beyond the third year of follow-up. Isolated locoregional or distant recurrence was observed in 52 and 20 patients, respectively, without significant difference in median time to relapse. 19 out of 43 recurrences with reported method of detection of relapse were diagnosed by routine technical investigations. There was no significant difference in time to relapse between those patients followed with low volume as compared to high-volume imaging protocols. CONCLUSIONS: Our data suggest that technical investigations in posttreatment surveillance should be restricted to the first 3 years of follow-up. Furthermore, surveillance programs with a high volume of imaging apparently do not lead to earlier detection or less advanced stage at the time of relapse as compared to protocols with low volume imaging. PMID- 19680689 TI - In vitro efficacy of ByeMite and Mite-Stop on developmental stages of the red chicken mite Dermanyssus gallinae. AB - The present in vitro study shows the efficacy of two antimite products (ByeMite = phoxim, Mite-Stop = neem seed extract) against all developing stages of the important red chicken mite Dermanyssus gallinae (obtained at two farms in France and Germany). While permanent contact with the active compound led to an efficacy of 100% in the case of Mite-Stop on mites in both farms, there was only a 96.2% killing effect of ByeMite on the mites of the French farm. Even short contacts of only 4 s killed 100% of mites in the case of Mite-Stop at the French farm and only 84.5% in the German farm. ByeMite, on the other hand, killed only 27.8% (Germany) and 30% (France) when mites got the chance to escape from the treated grounds to untreated ones. When using only the half doses of both products, Mite Stop(R) still reached, after permanent contact, 100% activity on the German farm and 98.2% in France, while ByeMite killed 93.8% (Germany) and 90.6% (France). Short contact to half doses of course reduced the activity of both products (Mite Stop = 59.3% in France, 22.1% in Germany; ByeMite = 28.8% in France, 18.8% in Germany). With respect to the fumigant activity of the products, the strains of D. gallinae reacted differently. While Mite-Stop(R) showed a clear fumigant activity in the case of the German mites, this product did not affect the French mites by air distribution, neither did ByeMite in both cases. Therefore, mites have to come in contact with both products. Against Mite-Stop, there was apparently no resistance and low doses have high efficacy after even short contacts, which regularly occur in a treated stable, where mites have the chance to leave treated places to untreated hidden spots. PMID- 19680690 TI - Cloning and characterization of peroxiredoxin in Babesia bovis. AB - We have identified the 2-Cys peroxiredoxin (Prx) from a bovine Babesia parasite, B. bovis. Prx is a recently described family of antioxidant enzymes that are highly conserved in eukaryotes and prokaryotes. B. bovis 2-Cys Prx (BbTPx-1) contained two conserved cysteine residues that corresponded to Cys47 and Cys170 of the yeast Prx and the amino acid sequences of two catalytic domains showed significant similarities to those of mammalian typical 2-Cys Prx. The antioxidant activity of the recombinant BbTPx-1 protein expressed in E. coli was demonstrated by a thiol mixed-function oxidation assay. Furthermore, we confirmed that BbTPx-1 was expressed in the cytoplasm of intra-erythrocytic B. bovis merozites. These results suggest that B. bovis likely uses TPx-1 as a way to reduce peroxides as a control of its intracellular redox balance so that it can live and grow in the host cell. PMID- 19680691 TI - Molecular characterization of hard ticks from Romania by sequences of the internal transcribed spacers of ribosomal DNA. PMID- 19680693 TI - Optimization of lipase-catalyzed glucose ester synthesis in ionic liquids. AB - Lipase-catalyzed esterification of glucose with fatty acids in ionic liquids (ILs) mixture was investigated by using supersaturated glucose solution. The effect of ILs mixture ratio, substrate ratio, lipase content, and temperature on the activity and stability of lipase was also studied. The highest yield of sugar ester was obtained in a mixture of 1-butyl-3-methylimidazolium trifluoromethanesulfonate ([Bmim][TfO]) and 1-methyl-3-octylimidazolium bis[(trifluoromethyl)-sulfonyl]amide ([Omim][Tf(2)N]) with a volume ratio of 9:1, while Novozym 435 (Candida antarctica type B lipase immobilized on acrylic resin) showed the optimal stability and activity in a mixture of [Bmim][TfO] and [Omim][Tf(2)N] with a 1:1 volume ratio. Reuse of lipase and ILs was successfully carried out at the optimized reaction conditions. After 5 times reuse of Novozym 435 and ILs, 78% of initial activity was remained. PMID- 19680694 TI - Enzymatic synthesis of glutathione using yeast cells in two-stage reaction. AB - In the present study, permeated yeast cells were used as the catalyst to synthesize glutathione. When waste cells of brewer's yeast were incubated with the three precursor amino acids and glucose for 36 h, 899 mg/L of glutathione were produced. To release the feedback inhibition of gamma-glutamylcysteine synthetase caused by glutathione, two-stage reaction was adopted. In the first stage, glycine was omitted from the reaction mixture and only gamma glutamylcysteine was formed. Glycine was then added in the second stage, and 1,569 mg/L of glutathione were produced. The conditions of the two-stage reaction were optimized using Plackett-Burman design and response surface methodology. Under the optimized condition, commercially available baker's yeast produced 3,440 mg/L of glutathione in 30 h, and most of the produced glutathione was in the medium. The two-stage reaction could effectively reduce the feedback inhibition caused by glutathione, but degradation of glutathione was significant. PMID- 19680692 TI - The junctions that don't fit the scheme: special symmetrical cell-cell junctions of their own kind. AB - Immunocytochemical, electron-, and immunoelectron-microscopical studies have revealed that, in addition to the four major "textbook categories" of cell-cell junctions (gap junctions, tight junctions, adherens junctions, and desmosomes), a broad range of other junctions exists, such as the tiny puncta adhaerentia minima, the taproot junctions (manubria adhaerentia), the plakophilin-2 containing adherens junctions of mesenchymal or mesenchymally derived cell types including malignantly transformed cells, the composite junctions (areae compositae) of the mature mammalian myocardium, the cortex adhaerens of the eye lens, the interdesmosomal "sandwich" or "stud" junctions in the subapical layers of stratified epithelia and the tumors derived therefrom, and the complexus adhaerentes of the endothelial and virgultar cells of the lymph node sinus. On the basis of their sizes and shapes, other morphological criteria, and their specific molecular ensembles, these junctions and the genes that encode them cannot be subsumed under one of the major categories mentioned above but represent special structures in their own right, appear to serve special functions, and can give rise to specific pathological disorders. PMID- 19680695 TI - Metabolic modelling of syntrophic-like growth of a 1,3-propanediol producer, Clostridium butyricum, and a methanogenic archeon, Methanosarcina mazei, under anaerobic conditions. AB - Clostridium butyricum can convert glycerol into 1,3-propanediol, thereby generating unfortunately a high amount of acetate, formate and butyrate as inhibiting by-products. We have proposed a novel mixed culture comprising C. butyricum and a methane bacterium, Methanosarcina mazei, to relieve the inhibition and to utilise the by-products for energy production. In order to examine the efficiency of such a mixed culture, metabolic modelling of the culture system was performed in this work. The metabolic networks for the organisms were reconstructed from genomic and physiological data. Several scenarios were analysed to examine the preference of M. mazei in scavenging acetate and formate under conditions of different substrate availability, including methanol as a co-substrate, since it may exist in glycerol solution from biodiesel production. The calculations revealed that if methanol is present, the methane production can increase by 130%. M. mazei can scavenge over 70% of the acetate secreted by C. butyricum. PMID- 19680696 TI - Resolution of sleep-disordered breathing in a dialysis-dependent child post-renal transplantation. AB - This case alerts paediatricians and renal physicians to the potential for significant sleep-disordered breathing in children with renal disease, particularly those with end stage kidney disease requiring dialysis. In some cases, management of the underlying renal disease may result in amelioration of the sleep-disordered breathing. Proactive sleep history taking and formal sleep monitoring in experienced centres may be indicated for these children to limit morbidity, especially if respiratory support is indicated. PMID- 19680697 TI - Clinical effects of nanocrystalline hydroxyapatite paste in the treatment of intrabony periodontal defects: a randomized controlled clinical study. AB - The purpose of the present randomized controlled clinical study was to compare the clinical outcomes of papilla preservation flap surgery with or without the application of a novel nanocrystalline hydroxyapatite (nano-HA) bone graft substitute. Fourteen patients with paired intrabony periodontal defects of >= 4 mm participated in this split-mouth design study. The defects in each subject were randomly selected to receive nano-HA paste in conjunction with papilla preservation flaps or papilla preservation flaps alone. Probing bone levels (PBL) from a customized acrylic stent and probing pocket depths (PPD) were measured at baseline and again 6 months following surgery. No differences in any of the investigated parameters were observed at baseline between the two groups. Healing was uneventful in all patients. Both treatments resulted in significant improvements between baseline and 6 months (p < 0.05). At 6 months after therapy, the sites treated with nano-HA paste showed a reduction in mean PPD from 8.3 +/- 1.2 to 4.0 +/- 1.1 mm and a gain in PBL of 4.3 +/- 1.4 mm, whereas in the control group, the mean PPD changed from 7.9 +/- 1.2 mm to 5.0 +/- 1.2 mm and PBL gain was 2.6 +/- 1.4 mm. Results demonstrated statistically greater PPD reduction and PBL gain (p < 0.05) in the test group as compared with the control group. In conclusion, after 6 months, the treatment of intrabony periodontal defects with a nano-HA paste leads to significantly improved clinical outcomes when compared with papilla preservation flap surgery alone. PMID- 19680698 TI - "Fatigue" having a role in the pathogenesis of osteonecrosis of the jaws. PMID- 19680699 TI - Tool-composite reuse in wild chimpanzees (Pan troglodytes): archaeologically invisible steps in the technological evolution of early hominins? AB - Recent etho-archaeological studies of stone-tool use by wild chimpanzees have contributed valuable data towards elucidating the variables that influenced the emergence and development of the first lithic industries among Plio-Pleistocene hominins. Such data help to identify potential behaviours entailed in the first percussive technologies that are invisible in archaeological records. The long term research site of Bossou in Guinea features a unique chimpanzee community whose members systematically use portable stones as hammers and anvils to crack open nuts in natural as well as in field experimental settings. Here we present the first analysis of repeated reuse of the same tool-composites in wild chimpanzees. Data collected over 5 years of experimental nut-cracking sessions at an "outdoor laboratory" site were assessed for the existence of systematic patterns in the selection of tool-composites, at group and at individual levels. Chimpanzees combined certain stones as hammer and anvil more often than expected by chance, even when taking into account preferences for individual stones by themselves. This may reflect an ability to recognise the nut-cracker as a single tool (composed of two elements, but functional only as a whole), as well as discrimination of tool quality-effectiveness. Through repeatedly combining the same pairs of stones--whether due to preferences for particular composites or for the two elements independently--tool-users may amplify use-wear traces and increase the likelihood of fracturing the stones, and thus of detaching pieces by battering. PMID- 19680700 TI - Eliciting health state utilities from the general public for severe chronic pain. AB - OBJECTIVE: There is an absence of data on public preferences for health states (HSs) associated with severe chronic pain. The aim of this study was to develop accurate HS descriptions for severe chronic pain requiring intrathecal (IT) therapy and to derive utility weights that describe the health-related quality of life (HRQL) impact of chronic pain associated with malignant (MP) and non malignant (NMP) aetiologies. RESEARCH DESIGN AND METHOD: Eight visual analogue scale pain index (VAS-PI) HSs were defined using ranges 0-40, 41-60, 61-80 and 81 100 applied to both MP and NMP. Additionally, eight HSs representing common adverse events associated with IT therapy were identified. The content and description of the HSs were ascertained by interviews with five United Kingdom clinical experts. In total, 16 HSs were compiled. These HS descriptions and HS questionnaires were administered to 102 members of the public, utilising a time trade off (TTO) approach to estimate utilities for the HSs. RESULTS: Participants generally were well matched to the general public in England and Wales, with some differences in mean age, race and education. A substantial decline in utility was observed with more severe VAS-PI values. The mean TTO utility values also decreased from mild pain to severe pain. Participants were able to differentiate between the side effects. CONCLUSION: The study shows a clear decrement in utility moving from different severity levels of severe chronic pain. PMID- 19680701 TI - Contribution of tissue composition and structure to mechanical response of articular cartilage under different loading geometries and strain rates. AB - Mechanical function of articular cartilage in joints between articulating bones is dependent on the composition and structure of the tissue. The mechanical properties of articular cartilage are traditionally tested in compression using one of the three loading geometries, i.e., confined compression, unconfined compression or indentation. The aim of this study was to utilize a composition based finite element model in combination with a fractional factorial design to determine the importance of different cartilage constituents in the mechanical response of the tissue, and to compare the importance of the tissue constituents with different loading geometries and loading rates. The evaluated parameters included water and collagen fraction as well as fixed charge density on cartilage surface and their slope over the tissue thickness. The thicknesses of superficial and middle zones, as based on the collagen orientation, were also included in the evaluated parameters. A three-level resolution V fractional factorial design was used. The model results showed that inhomogeneous composition plays only a minor role in indentation, though that role becomes more significant in confined compression and unconfined compression. In contrast, the collagen architecture and content had a more profound role in indentation than with two other loading geometries. These differences in the mechanical role of composition and structure between the loading geometries were emphasized at higher loading rates. These findings highlight how the results from mechanical tests of articular cartilage under different loading conditions are dependent upon tissue composition and structure. PMID- 19680702 TI - Deformation-induced hydrolysis of a degradable polymeric cylindrical annulus. AB - A thermodynamically consistent framework for describing the response of materials undergoing deformation-induced degradation is developed and applied to a particular biodegradable polymer system. In the current case, energy is dissipated through the mechanism of hydrolytic degradation and its effects are incorporated in the constitutive model by appropriately stipulating the forms for the rate of dissipation and for the degradation-dependent Helmholtz potential which changes with the extent of the degradation of the material. When degradation does not occur, the response of the material follows the response of a power-law generalized neo-Hookean material that fits the response of the non degraded poly(L: -lactic acid) under uniaxial extension. We study the inflation and extension of a degrading cylindrical annulus and the influence of the deformation on the mechanism of degradation and its consequent mechanical response. Depreciation of mechanical properties due to degradation confers time dependent characteristics to the response of the biodegradable material: the material creeps when subjected to constant loads and stresses necessary to keep a fixed deformation relax. PMID- 19680703 TI - Flavonoids and cognitive function: a review of human randomized controlled trial studies and recommendations for future studies. AB - Evidence in support of the neuroprotective effects of flavonoids has increased significantly in recent years, although to date much of this evidence has emerged from animal rather than human studies. Nonetheless, with a view to making recommendations for future good practice, we review 15 existing human dietary intervention studies that have examined the effects of particular types of flavonoid on cognitive performance. The studies employed a total of 55 different cognitive tests covering a broad range of cognitive domains. Most studies incorporated at least one measure of executive function/working memory, with nine reporting significant improvements in performance as a function of flavonoid supplementation compared to a control group. However, some domains were overlooked completely (e.g. implicit memory, prospective memory), and for the most part there was little consistency in terms of the particular cognitive tests used making across study comparisons difficult. Furthermore, there was some confusion concerning what aspects of cognitive function particular tests were actually measuring. Overall, while initial results are encouraging, future studies need to pay careful attention when selecting cognitive measures, especially in terms of ensuring that tasks are actually sensitive enough to detect treatment effects. PMID- 19680704 TI - Lupin protein compared to casein lowers the LDL cholesterol:HDL cholesterol-ratio of hypercholesterolemic adults. AB - BACKGROUND: Lupin protein had hypocholesterolemic effects in laboratory animals. However, the effect in humans has not been elucidated till now. AIM OF THE STUDY: To investigate the effect of lupin protein on circulating cholesterol in plasma and lipoproteins of hypercholesterolemic subjects. SUBJECTS AND METHODS: A randomised, double-blind, placebo-controlled, parallel trial (23 females and 20 males completed the trial) was conducted to compare the effects of lupin protein versus casein as control protein on plasma lipids and amino acids. Thirty-five grams of the test protein were consumed daily for 6 weeks. RESULTS: Both lupin protein and casein resulted in a reduction of circulating plasma cholesterol ( 0.50 +/- 0.64 and -0.47 +/- 0.79 mM; P < 0.05) from baseline to week 6. The reduction of plasma cholesterol was mainly caused by a reduction of LDL cholesterol in the lupin protein group (-0.31 +/- 0.46 mM; P < 0.05), while in the casein group HDL cholesterol significantly declined (-0.17 +/- 0.15 mM; P < 0.05). Comparing the lupin protein group with the casein group yielded a difference in the net changes from baseline to week 6 in the LDL:HDL cholesterol ratio of -0.24 (95% CI: -0.007, -0.479; P < 0.05). No significant differences in net changes were observed for plasma concentrations of triglycerides, glucose, homocysteine, taurine and most of the amino acids. CONCLUSIONS: Lupin protein compared to casein slightly lowered the concentration of LDL cholesterol in hypercholesterolemic subjects, without altering HDL cholesterol. No or minor effects of lupin protein were observed on circulating glucose, homocysteine and plasma amino acids. PMID- 19680705 TI - Effect of water soluble vitamins on Zn transport of Caco-2 cells and their implications under oxidative stress conditions. AB - BACKGROUND: The role of different water soluble vitamins in Zn metabolism beyond intestinal Zn absorption is poorly explored. AIM OF THE STUDY: Using Caco-2 cells, effects of different vitamins on intestinal Zn transport and their implications under oxidative stress (OS) were investigated. METHODS: Cells were apically treated with Zn (25 muM) and vitamins (Folic acid (FA), Nicotinic acid (NA), Ascorbic acid (AA), riboflavin, thiamine, pyridoxine) for 60 min. The effect of most promising vitamins on zinc transport, antioxidant enzymes (Catalase, Glutathione peroxidase, and superoxide dismutase), and intracellular OS status (ROS generation and mitochondrial transmembrane potential) were investigated. OS was generated by tert-butyl hydro peroxide and results for each vitamin were compared with respective Zn containing controls with and without OS. RESULTS: Without OS, Zn transport was slightly enhanced in presence of NA, while it was significantly reduced by thiamine, riboflavin, and pyridoxine. Under OS, NA significantly (P < 0.01) enhanced Zn transport in dose-dependent manner, while, pyridoxine and AA moderately improved it. Under both conditions, Zn transport exhibited decreasing trend with increase of FA. The antioxidant enzyme and OS markers levels varied significantly in Zn + vitamins. With Zn + FA + OS, enzyme activities decreased maximally, with twofold increase in 2',7' dichlorofluorescin diacetate (DCF-DA) (P < 0.01) and lowering of rhodamine fluorescence (P < 0.05). In Zn + AA + OS, DCF-DA fluorescence increased (P < 0.05) but with NA, cellular enzymes, and antioxidant profile were improved. CONCLUSIONS: Results for the first time demonstrate advantageous effects of NA and deleterious consequences of FA with no effect by AA on Zn transport, especially under OS. These observed changes in the transport of Zn seem to have an impact on OS markers. PMID- 19680706 TI - Influence of repetitive transcranial magnetic stimulation on psychomotor symptoms in major depression. AB - Psychomotor symptoms related to an impairment of the nigrostriatal dopaminergic system are frequent in major depression (MD). Repetitive transcranial magnetic stimulation (rTMS) has been discussed as a new treatment option for MD. In neurobiological terms, an influence of high-frequency rTMS on dopaminergic neurotransmission has previously been shown by several studies in animals and humans. Therefore, an improvement of psychomotor symptoms by rTMS could be assumed. The aim of this pilot study was to investigate the effect of high frequency rTMS on psychomotor retardation and agitation in depressive patients. We investigated the effect of left prefrontal 10 Hz rTMS on psychomotor retardation and agitation in 30 patients with MD. Patients were randomly assigned to real or sham rTMS in addition to a newly initiated standardized antidepressant medication. We found a trend in the reduction of agitation (t(28) = 1.76, p = 0.09, two-tailed), but not in the reduction of retardation. Furthermore, no general additional antidepressant effect of rTMS was observed. Although there was no statistical significant influence of high-frequency rTMS on psychomotor symptoms in depressive patients, the results showed a trend in the reduction of psychomotor agitation in MD. This effect should be systematically investigated as the primary end point in further studies with larger sample sizes. PMID- 19680707 TI - [Changes in labor market participation of older employees in Germany: the perspective of labor market research]. AB - For many years, Germany has been regarded in international comparisons as an example of a generous early retirement culture, resulting in a low labor market participation of older employees. Recently, however, employment rates of older employees have increased remarkably. Reasons are the demographic structure of older persons in Germany, a long-term trend of increasing female labor market participation, and reforms in labor-market policies and pension policies during the last 10 years. Despite an increasing labor market participation of older employees, traditional labor market risks for older persons partly remained, but some new risks evolved as well. Therefore, social differentiation among older employees increased.Although detailed macro descriptions exist, the causes of labor market developments cannot be fully understood with cross-sectional data alone. An important stimulus is to be expected from individual longitudinal data which reflect employment histories and labor market transitions such as employment exit and retirement. PMID- 19680708 TI - [Age management in operational practice: drivers, implementation strategies, and effects]. AB - Against the background of expected changes in demography, this paper takes a deeper look at the concept of organizational age management. On the basis of the analysis of 32 organizational examples of good practice in age management, this paper seeks to make statements about the drivers, implementation strategies and effects of age management and derive requirements from this. PMID- 19680709 TI - [Cognitive competence of older workers]. AB - The article provides a short overview on cognitive strengths and weaknesses of older workers. The older people who are under the age of 65 years already exhibit changes in performance and neurophysiological measures in laboratory tasks that test fluid intelligence. Apart from deficits, the physiological data of the older people show clear evidence for compensatory strategies. Cognitive performance on the current job is usually less impaired. This can be explained by practice, learning, and selection, which are, however, often linked with higher effort. The cognitive competence of older employees varies exorbitantly, which is due to the influence of various factors like education, lifestyle, and in particular the type of work. Thus, the cognitive competence of older employees can be preserved and possibly even enhanced by changes in the work situation as well as by individual training procedures such as cognitive training. PMID- 19680710 TI - [Post-retirement activity in Germany]. AB - During recent years, a discussion on work activity of retirees has commenced also in Germany. First, this contribution presents a survey of the North American phenomenon of bridge employment and, second, complements leadoff empirical results on individual motives and favored basic conditions in Germany. Individual and structural preconditions for work activity during retirement are presented as well as possible manifestations of retirement activities. This paper quantifies the extent of work activity of retirees in Germany into their upper seventies. Potential follow-up actions for the three levels individuals, organizations, as well as politics and society are recommended. PMID- 19680712 TI - [Communication of the Federal Association of Geriatrics]. PMID- 19680711 TI - [Communication of the German Society of Gerontology and Geriatrics (DGGG)]. PMID- 19680714 TI - Abstracts of the 57th Annual Scientific Meeting of the Irish Gerontological Society. Belfast, Ireland. September 25-26, 2009. PMID- 19680713 TI - Bone healing of the sheep tibia shaft after carbon dioxide laser osteotomy: histological results. AB - The aim of the study was to compare the histological results after complete osteotomies of the sheep tibia using either the prototype carbon dioxide (CO(2)) laser osteotome 'OsteoLAS' (n = 12) or an oscillating saw (n = 12). The laser parameters were as follows: wavelength 10.6 microm; energy of laser pulses 75-85 mJ; pulse duration 80 mus; pulse repetition rate 200 Hz; spot diameter 460 mum (1/e(2) level); radiant exposure 45-51 J/cm(2); peak irradiance 0.56-0.64 MW/cm(2). Both groups were divided into two subgroups (n = 6), and the animals were killed after 4 weeks or 12 weeks, respectively. Light and fluorescence microscopy with semiquantitative analysis and histomorphometry were performed to compare bone healing. Charring-free laser osteotomies were possible up to a depth of 20 mm with the short-pulsed CO(2) laser. The laser, however, required a significantly longer time to perform, and a wedge-shaped gap was present on the cis-cortex. After 4 weeks the osteotomy gaps were almost unchanged in both groups and filled with connective tissue. After 12 weeks the gaps were filled with newly formed bone in both groups. Primary gap healing was predominant in the laser group and longitudinal cortical remodelling in the control group. On a cellular level, no fundamental differences were observed for early and late stages of bone healing. Further research has to be focussed on improving the CO(2) laser ostetome in order to reduce the long duration of the laser osteotomy and the necessity of creating a wedge-shaped cut in thick bones. PMID- 19680715 TI - Abstracts of the 34th Sir Peter Freyer Memorial Lecture and Surgical Symposium. September 4-5, 2009. Galway, Ireland. PMID- 19680716 TI - Management of arterial hypertension occurring early after living donor liver transplantation in children: report of three cases and review of the literature. AB - Three pediatric patients with hypertension occurring early after liver transplantation are reviewed. The patients were all female, and underwent living donor liver transplantation at the age of 9 years, 1 month, and 7 months. The etiology of liver disease was cirrhosis due to biliary atresia in two patients and fulminant hepatitis in one patient. Antihypertensive therapy with calcium channel blocker alone was not effective. Blood pressure was eventually controlled after the administration of a beta-adrenergic blocker in addition to the calcium channel blocker to all patients. No end-organ damage was observed, except that two patients developed temporary left ventricular hypertrophy. PMID- 19680717 TI - Modeling the impacts of two bark beetle species under a warming climate in the southwestern USA: Ecological and economic consequences. AB - Predicted climate warming is expected to have profound effects on bark beetle population dynamics in the southwestern United States. Temperature-mediated effects may include increases in developmental rates, generations per year, and changes in habitat suitability. As a result, the impacts of Dendroctonus frontalis and Dendroctonus mexicanus on forest resources are likely subject to amplification. To assess the implications of such change, we evaluated the generations per year of these species under three climate scenarios using a degree-day development model. We also assessed economic impacts of increased beetle outbreaks in terms of the costs of application of preventative silvicultural treatments and potential economic revenues forgone. Across the southwestern USA, the potential number of beetle generations per year ranged from 1-3+ under historical climate, an increase of 2-4+ under the minimal warming scenario and 3-5+ under the greatest warming scenario. Economic benefits of applying basal area reduction treatments to reduce forest susceptibility to beetle outbreaks ranged from $7.75/ha (NM) to $95.69/ha (AZ) under historical conditions, and $47.96/ha (NM) to $174.58/ha (AZ) under simulated severe drought conditions. Basal area reduction treatments that reduce forest susceptibility to beetle outbreak result in higher net present values than no action scenarios. Coupled with other deleterious consequences associated with beetle outbreaks, such as increased wildfires, the results suggest that forest thinning treatments play a useful role in a period of climate warming. PMID- 19680718 TI - Compression of the inferior vena cava by the right iliac artery: a rare variant of May-Thurner syndrome. AB - May-Thurner syndrome is known as compression of the left common iliac vein by the right common iliac artery. We describe a case of an atypical compression of the inferior vena cava by the right common iliac artery secondary to a high aortic bifurcation. Despite an extensive collateral network, there was a significant venous gradient between the iliac veins and the inferior vena cava above the compression. After stenting the venous pressure gradient disappeared. Follow-up 4 months later revealed a good clinical response with a patent stent. PMID- 19680719 TI - Rhabdomyolysis developing after transcatheter arterial chemoembolization for hepatocellular carcinoma. AB - A 25-year-old man with hepatocellular carcinoma developed severe muscular weakness and pain 15 days after transcatheter arterial chemoembolization (TACE). The diagnosis of rhabdomyolysis was made based on myalgia localized in the bilateral upper extremities (bilateral trapezius, deltoid, biceps brachii, and teres major muscles) on magnetic resonance imaging and increased levels of muscle derived serum enzymes. In this case, some drugs administered during the clinical course of TACE (diclofenac, famotidine, and cefotiam dihydrochloride) were suspected to be involved in the rhabdomyolysis, but the exact cause of rhabdomyolysis was not identified. The symptoms were completely improved by right trisegmentectomy of the liver following conservative treatment. PMID- 19680720 TI - Selective internal radiation therapy with SIR-spheres for the treatment of unresectable colorectal hepatic metastases. AB - The purpose of this study was to evaluate the effectiveness of colorectal cancer (CRC) liver metastasis radioembolization with yttrium-90 (Y90), assessing toxicity and survival rates in patients with no response to chemotherapy through our 3-year experience. From February 2005 to January 2008, we treated 41 patients affected by CRC from a cohort of selective internal radiation therapy patients treated at our institution. All patients examined showed disease progression and arrived for our observation with an abdominal CT, a body PET, and a hepatic angiography followed by gastroduodenal artery coiling previously performed by us. We excluded patients with a bilirubin level>1.8 mg/dl and pulmonary shunt>20% but not patients with minor extrahepatic metastases. On treatment day, under fluoroscopic guidance, we implanted a dose of Y90 microspheres calculated on the basis of liver tumoral involvement and the body surface area formula. All patients were discharged the day after treatment. We obtained, according to Response Evaluation Criteria on Solid Tumors, a complete response in 2 patients, a partial response in 17 patients, stable disease in 14 patients, and progressive disease in 8 patients. In all cases, we obtained a carcinoembryonic antigen level decrease, especially in the week 8 evaluation. Technical success rate was 98% and technical effectiveness estimated at 3 months after treatment was 80.5%. Side effects graded by Common Terminology Criteria on Adverse Events were represented by one grade 4 hepatic failure, two grade 2 gastritis, and one grade 2 cholecystitis. The median survival and the progression-free survival calculated by Kaplan-Meier analysis were 354 and 279 days, respectively. In conclusion, according to our 3-year experience, Y90 SIR-Spheres radioembolization is a feasible and safe method to treat CRC liver metastases, with an acceptable level of complications and a good response rate. PMID- 19680721 TI - Bacterial diversity and bioprospecting for cold-active lipases, amylases and proteases, from culturable bacteria of kongsfjorden and Ny-alesund, Svalbard, Arctic. AB - Culturable bacterial diversity of seven marine sediment samples of Kongsfjorden and a sediment and a soil sample from Ny-Alesund, Svalbard, Arctic was studied. The bacterial abundance in the marine sediments of Kongsfjorden varied marginally (0.5 x 10(3)-1.3 x 10(4) cfu/g sediment) and the bacterial number in the two samples collected from the shore of Ny-Alesund also was very similar (0.6 x 10(4) and 3.4 x 10(4), respectively). From the nine samples a total of 103 bacterial isolates were obtained and these isolates could be grouped in to 47 phylotypes based on the 16S rRNA gene sequence belonging to 4 phyla namely Actinobacteria, Bacilli, Bacteroidetes and Proteobacteria. Representatives of the 47 phylotypes varied in their growth temperature range (4-37 degrees C), in their tolerance to NaCl (0.3-2 M NaCl) and growth pH range (2-11). Representatives of 26 phylotypes exhibited amylase and lipase activity either at 5 or 20 degrees C or at both the temperatures. A few of the representatives exhibited amylase and/or lipase activity only at 5 degrees C. None of the phylotypes exhibited protease activity. Most of the phylotypes (38) were pigmented. Fatty acid profile studies indicated that short chain fatty acids, unsaturated fatty acids, branched fatty acids, the cyclic and the cis fatty acids are predominant in the psychrophilic bacteria. PMID- 19680722 TI - The C-terminal domain of BinA is responsible for Bacillus sphaericus binary toxin BinA-BinB interaction. AB - The binary toxin (Bin) from Bacillus sphaericus consists of two polypeptides, BinA (42 kDa) and BinB (51 kDa) that work together to kill susceptible mosquito larvae. To investigate the functional regions of BinA involved in the interaction with BinB, four BinA truncated fragments, from both N- and C- termini, were constructed and expressed in Escherichia coli. Neither individual nor a mixture of fragments of BinA showed larvicidal activity against Culex quinquefasciatus larvae even using a high dose of toxins. Far-Western dot blot analysis showed strong binding of both C-terminal fragments (17 and 28 kDa) to BinB protein. This is the first report to demonstrate that the C-terminal part of BinA plays an important role for the interaction with BinB. PMID- 19680723 TI - The impact of dysphagia on quality of life in ageing and Parkinson's disease as measured by the swallowing quality of life (SWAL-QOL) questionnaire. AB - This prospective, cross-sectional study evaluated the impact of dysphagia on quality of life in healthy ageing and in subjects with Parkinson's disease (PD) using the Swallowing Quality of Life (SWAL-QOL) questionnaire. Sixteen healthy young adults (8 males, mean age = 25.1 years) and 16 healthy elders (8 males, mean age = 72.8 years) were recruited. Thirty-two subjects with idiopathic PD (mean age = 68.5 years) were recruited from a movement disorders clinic. The severity of PD was staged using the Hoehn and Yahr scale. Results revealed that elders experienced symptoms of dysphagia more frequently than young adults but the overall SWAL-QOL scores were not significantly different. Subjects with PD who experienced dysphagia reported greatly reduced QOL, and significant differences were found in all but one subsection of the SWAL-QOL. Disease progression detrimentally impacts QOL, with subjects in later-stage PD experiencing further reduction in the desire to eat, difficulty with food selection, and prolonged eating duration. These features, which increase with disease severity, are likely to impact negatively upon nutritional status, which is already under threat from PD-related dysphagia. PMID- 19680724 TI - Sensory and motor responses of normal young adults during swallowing of foods with different properties and volumes. AB - We examined the influence of rheological/textural properties and volumes of test foods on the sensory and motor aspects of swallowing in healthy young adults. Three test foods differing in thickening agent concentration (0.0, 1.5, and 3.0%) were prepared and delivered in different volumes (approximately 3, approximately 5, and approximately 7 ml) to subjects seated on a chair. Viscosity analyses of the 1.5 and 3.0% test foods revealed that they behaved as non-Newtonian fluids and were thixotropic. The 1.5% test food differed from the 3.0% test food in its textural properties (hardness, cohesiveness, and adhesiveness). As determined by a linear model equation method, the thickening agent concentration affected the scores of all six sensory evaluation questions that were answered by the subjects, which suggests that the concentration affected the food properties being evaluated. Consistent with previous reports, thickening agent concentration and test food volume also affected some durational parameters of laryngeal (recorded by a piezoelectric sensor) and suprahyoid muscle (recorded on an electromyogram) motor activity. However, thickening agent concentration and test food volume did not affect the single amplitude parameter of the electromyogram that was measured. The thixotropic property of foods can affect the motor aspect of oropharyngeal swallowing as well as the sensory aspect. PMID- 19680725 TI - 1-(3',5'-dihydroxyphenoxy)-7-(2'',4'',6-trihydroxyphenoxy)-2,4,9 trihydroxydibenzo-1,4-dioxin inhibits adipocyte differentiation of 3T3-L1 fibroblasts. AB - In this study, we isolated the phloroglucinol derivative, 1-(3',5' dihydroxyphenoxy)-7-(2'',4'',6-trihydroxyphenoxy)-2,4,9-trihydroxydibenzo-1,4 dioxin (1), from Ecklonia cava and evaluated its potential inhibition on adipocyte differentiation in 3T3-L1 cells. Lipid accumulation along with the expression of several genes associated with adipogenesis and lipolysis was examined at the end of differentiation. Lipid accumulation level was examined by measuring triglyceride content and Oil-Red O staining. The expression levels of several genes and proteins were examined using reverse-transcription polymerase chain reaction (RT-PCR), real-time RT-PCR, and Western blot analysis. Compound 1 significantly reduced lipid accumulation and downregulated peroxisome proliferator-activated receptor-gamma, sterol regulatory element-binding protein 1c, and CCAAT/enhancer-binding proteins alpha in a dose-dependent manner. Moreover, the presence of compound 1 induced downregulation of adipogenic target genes such as fatty acid binding protein 4, fatty acid transport protein 1, fatty acid synthase, acyl-CoA synthetase 1, lipoprotein lipase, and leptin. According to the lipolytic response, compound 1 downregulated perilipin and hormone sensitive lipase while upregulating tumor necrosis factor alpha. Therefore, these results suggest that compound 1 might decrease lipid accumulation during adipocyte differentiation by modulating adipogenesis and lipogenesis. Furthermore, compound 1 could be developed as a functional agent effective in improving obesity. PMID- 19680726 TI - Comparative toxicity of mycotoxins to navel orangeworm (Amyelois transitella) and corn earworm (Helicoverpa zea). AB - Mycotoxins, such as aflatoxins and ochratoxins, are widely distributed in nature and are frequently problematic crop contaminants that cause millions of dollars of annual losses in the United States. Insect infestations of crop plants significantly exacerbate mycotoxin contamination. Damage to a variety of nut species by Amyelois transitella Walker (navel orangeworm, NOW) is associated with infection by Aspergillus species and concomitant production of aflatoxins and ochratoxins. Resistance to aflatoxins in this lepidopteran is compared here with the levels of resistance in Helicoverpa zea (corn earworm, CEW), another lepidopteran that routinely encounters aflatoxins in its diet, albeit at lower levels. Measured as the developmental delay caused by aflatoxin B1 (AFB1), it is apparent that the LC(50) (defined as the concentration preventing 50% of newly hatched larvae from entering the 2nd instar within 48 h) for AFB1 is 100 times greater for A. transitella than for H. zea. Similarly, A. transitella 1st instars display substantially higher tolerance to ochratoxin A, another mycotoxin contaminant produced by Aspergillus species, than do H. zea. Our studies indicate that A. transitella, although a hostplant generalist, may well be highly specialized for mycotoxin detoxification. PMID- 19680727 TI - Use of percutaneous radiofrequency ablation in pulmonary metastases from renal cell carcinoma. AB - BACKGROUND: Surgical resection of pulmonary metastases from renal cell carcinoma (RCC) has been demonstrated in recent studies to produce good long-term survival outcomes. Radiofrequency ablation (RFA) may offer an alternative treatment option for this group of patients, especially for those who are unable to have surgery. METHODS: Nine patients had a total of 23 pulmonary metastases treated with percutaneous RFA under fluoro-computed tomography (CT) guidance. Patients underwent routine overnight hospitalization and monitoring for other potential complications. CT scans were performed at 1 month after the procedure and at 3 monthly intervals. RESULTS: A total of 25 ablations were performed to 23 pulmonary metastases for our nine patients in 12 RFA sessions. No patient died within 30 days of the procedure. Five of the 12 procedures resulted in a pneumothorax (42%) and 3 required insertion of a Pleurocath. One patient had a bronchopulmonary fistula with an associated small pleural effusion and one patient had pneumonia. Of the 25 ablations, 14 lesions had decreased in size (56%), 1 was stable in size (4%), and 9 had increased in size (36%). One patient had deceased before adequate follow-up. Of the nine patients, two are alive and free of disease (mean survival time of 74 months), two are alive with disease (mean survival time of 16 months), and five have died of disease (mean survival time of 26.2 months). CONCLUSION: RFA offers a treatment alternative for local tumor control, particularly in selected patients with multiple, small lesions who are unsuitable for pulmonary resections. PMID- 19680728 TI - Urologic complications of composite resection following combined modality treatment of colorectal cancer. AB - BACKGROUND: It is a common perception that preoperative radiation increases the incidence of urologic complications following composite resection, but there is little evidence to support or refute this claim. METHODS: Patients who underwent ureteric reconstruction as a component of en bloc resection for locally advanced or recurrent colorectal cancer were identified from a multicenter institutional database (1982-2007). Charts were reviewed to determine the incidence, nature, management, and predictors of serious urologic complications. RESULTS: 126 patients (34 female, 92 male) met the inclusion criteria: (1) division of one or both ureters; and (2) ureteric reconstruction involving anastomosis to ureter, bladder, small bowel or colon. Urologic complications requiring intervention occurred in 30 (24%) patients, including: anastomotic leak (n = 11), anastomotic stricture (n = 10), fistula (n = 5), conduit/stoma problem (n = 5), and other (n = 3). Eight patients required surgical revision. The rate of urologic complications was similar in patients who had received radiation at some time prior to the index surgery and those who had not (25% and 19%, respectively, P = 0.61). There was a trend towards a higher complication rate with more complex compared with simpler reconstructive procedures (38% versus 20%, P = 0.069). CONCLUSION: 24% of patients who underwent ureteric reconstruction following composite resection developed a urologic complication that required intervention. Preoperative radiotherapy did not appear to predispose patients to urologic complications. Optimization of surgical technique at the time of en bloc resection may obviate the need for subsequent revision. PMID- 19680729 TI - Intra-individual comparison of lymphatic drainage patterns using subareolar and peritumoral isotope injection for breast cancer. AB - BACKGROUND: Controversy exists in the literature regarding the optimal site for lymphatic mapping in breast cancer. This study was designed to characterize lymphatic drainage patterns within the same patient after subareolar (SA) and peritumoral (PT) radiopharmaceutical injections and examine the impact of reader interpretation on reported drainage. METHODS: In this prospective trial, 27 women with breast cancer underwent sequential preoperative SA and PT injections of 0.5 to 2.7 mCi of technetium-99 m filtered sulfur colloid 3 days or more apart. Patterns of radiopharmaceutical uptake were reviewed independently by two nuclear medicine physicians. Inter-reader agreement and injection success were assessed in conjunction with observed drainage patterns. RESULTS: There was near perfect inter-reader agreement observed on identification of axillary LN drainage after PT injection (P = 0.0004) and substantial agreement with SA injection (P = 0.0344). SA injection was more likely to drain to only axillary LNs, whereas PT injection appeared more likely to drain to both axillary and extra-axillary LNs, although no statistically significant differences were found. All patients with extra-axillary drainage after PT injection (n = 6 patients) had only axillary drainage after SA injection. Dual drainage was observed for six patients with PT injection and one patient with SA injection. CONCLUSIONS: Our findings suggest that radiopharmaceutical injected in the SA location has a high propensity to drain to axillary LNs only. After controlling for patient factors and demonstrating inter-reader agreement, the inability to demonstrate statistically significant differences in drainage based on injection site suggests that lymphatic drainage patterns may be a function of patient and tumor-specific features. PMID- 19680730 TI - Weight loss and quality of life after gastric band removal or deflation. AB - BACKGROUND: The study aim was to retrospectively assess whether patients were able to maintain their weight after gastric band removal or deflation and how they felt about gastric banding. METHODS: A total of 41 patients (93% female, mean age 34.1 (SD 10.5) years) were included in this study: patients who had their band removed/deflated without further surgical intervention (group 1, n = 26) and those who later underwent a second bariatric operation (group 2, n = 15). We evaluated weight gain after band removal/during the time between band removal and second bariatric operation. RESULTS: Of our patients, 31 (76%) suffered a complication (18 late pouch dilatations, six band infections, five band migrations, and two band leaks) requiring band removal. Ten patients wanted their band removed (six) or emptied (four). Mean time after band removal, when patients had neither a band nor a second bariatric operation, was 2.84 (SD 2.3) years. Five (12.2%) patients maintained their weight, four of whom experienced a learning effect; all others gained weight. Mean body mass index for both groups after the period without a band was 36.7 (SD 8.0) kg/m(2) (vs 29.4 (SD 7.0) at removal), and excess weight loss was 33.2% (SD 39.2; vs 69.8% (SD 32.9) at removal). Of our patients, 73% would not agree to gastric banding again. According to the bariatric analysis and reporting outcome system, long-term outcome of patients following band removal was a "failure" in 66% of patients. CONCLUSIONS: Long-term outcome following band removal is unsatisfactory in many patients. Nevertheless, a minority of patients was able to maintain its weight loss. PMID- 19680731 TI - When the brakes came off: re-feeding oedema after deflation of a gastric band: a case report. AB - Bariatric surgery is now the treatment of choice for morbid obesity, but is not without risk. Patients are cared for in specialised centres, but complications can present to non-specialised centres. We describe life-threatening re-feeding oedema in a patient following routine deflation of a gastric band. Band deflation or removal may be required for various reasons, but rapid release of the band without additional supplementation of electrolytes may be dangerous due to re feeding syndrome. PMID- 19680732 TI - Subfascial port placement in gastric banding surgery. AB - BACKGROUND: In some bariatric patients with predominantly intra-abdominal fat a shallow fat layer separates the gastric band access port from the skin. We hypothesise that subfascial port placement in these patients reduces skin erosions and port infections and improves cosmesis as weight loss occurs. AIM: This study aims to compare port complications, cosmetic outcome and ease of band adjustment with access ports in front of or behind the rectus muscle. METHOD: We retrospectively compared complications and cosmetic outcomes of patients with subfascial ports to a control group matched for gender, BMI and age. Each subject completed a questionnaire utilising a 1 to 10 scale for nine parameters related to comfort and cosmesis and two parameters related to discomfort during adjustments. RESULTS: Sixty-eight patients with subfascial ports were identified and the overall response rate was 84%. The groups were well matched for gender (m:f ratio 1.8:1 vs. 1.7:1, p = 1.000), age (51.0 vs. 49.6 years, p = 0.528) and BMI (39.8 vs. 40.3 kg/m2, p = 0.585). There was no difference in port infection rates (0/68 vs. 1/68, p = 1.000) but the subfascial group had more hernias (3/68 vs. 0/68, p = 0.244). Subfascial patients experienced more pain during adjustments (score 4.3 vs. 2.6, p = 0.047) but a combined analysis of cosmesis showed a slight positive trend (1.58 vs. 1.76, p = 0.379). CONCLUSION: Both port locations are well tolerated. Subfascial placement is associated with more pain during adjustments but there is no difference in port infection or skin erosion rates. PMID- 19680733 TI - Liver transplantation for acute-on-chronic liver failure. AB - PURPOSE: To evaluate the outcome of liver transplantation for acute-on-chronic liver failure. PATIENTS AND METHODS: From November 1991 to December 2007, 517 patients underwent liver transplantation at Queen Mary Hospital, Hong Kong. Among them, 149 had acute-on-chronic liver failure as defined in the recent Asian Pacific Association for the Study of Liver Consensus Meeting. Their clinical data were reviewed and their survival outcomes were compared with those of patients who underwent liver transplantation for fulminant hepatic failure and for cirrhosis only in the same period. RESULTS: The patients with acute-on-chronic liver failure included 50 patients having acute exacerbation of chronic hepatitis B and 99 cirrhotic patients with acute deterioration. Their median model for end stage liver disease scores were 35 and 37, respectively. Preoperative infection (35%), hepatorenal syndrome (38%), and respiratory failure (28.8%) were common. One hundred and three patients received living donor liver grafts and 46 patients received deceased donor liver grafts. The hospital mortality rate was 4.7%. The 5 year survival rates were 93.2% for patients with acute exacerbation of chronic hepatitis B and 90.5% for cirrhotic patients with acute deterioration. The results were similar to those of the patients with fulminant hepatic failure (n = 37) and the patients having cirrhosis only (n = 301). CONCLUSIONS: Liver transplantation for acute-on-chronic liver failure is life-saving, and the survival rates it attains are similar to those attained by transplantation for other liver conditions. PMID- 19680734 TI - Case report: absent C6 cervical pedicle in a collegiate football player. AB - Congenital absence of a cervical pedicle is a rare clinical finding with only 70 reported cases in the literature from 1946 until present. The congenitally absent pedicle has clinical importance owing to the frequency of misdiagnosis and inappropriate invasive treatments. We present the case of a 21-year-old college football player who experienced neck and shoulder pain after violent twisting of his neck by the face mask. The player walked off the field under his own power. He was sent to the locker room, where he underwent right shoulder and cervical spine radiographs. Initial review of the radiographs raised concern for a jumped right C6 facet. The patient then underwent CT and MRI of the cervical spine, confirming the diagnosis of an absent cervical pedicle. He was treated nonoperatively for a short time and completed the season. He had no symptoms at last followup at 8 months. The most frequent location of the absent cervical pedicle is at the C6 level, and the next most common is at the C5 level. Neural compression or instability is uncommon and nonsurgical treatment is the mainstay of treatment. Misdiagnosis can lead to inappropriate treatment such as halo or tong application with traction, which occurred in seven of 57 cases in one series, and exploratory surgery, which occurred in four of 57 cases. PMID- 19680735 TI - Complications of femoral nerve block for total knee arthroplasty. AB - Preemptive and multimodal pain control protocols have been introduced to enhance rehabilitation after total knee arthroplasty (TKA). We determined the complication rate associated with preoperative femoral nerve block (FNB) for TKA. Among 1018 TKA operations, we performed 709 FNBs using a single-injection technique into the femoral nerve sheath and confirming position with nerve stimulation before induction. After TKA, weightbearing as tolerated was initiated using a walker or crutches on postoperative Day 1. Twelve patients (1.6%) treated with FNB sustained falls, three (0.4%) of whom underwent reoperations. Five patients had postoperative femoral neuritis, which may have been secondary to the block. One patient had new onset of atrial fibrillation after FNB, and the TKA was postponed. Femoral nerve block before TKA is not a harmless intervention. We recommend postoperative protocols be modified for patients who have FNB to account for decreased quadriceps function in the early postoperative period, which can lead to falls. LEVEL OF EVIDENCE: Level III, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence. PMID- 19680736 TI - Hydrogen sulfide scavenges the cytotoxic lipid oxidation product 4-HNE. AB - Highly reactive alpha,beta-unsaturated aldehydes like 4-hydroxy-2-nonenal (4 HNE), generated from oxidation of polyunsaturated fatty acids, can bind to proteins, polynucleotides and exert cytotoxicity. 4-HNE is known to react readily with thiol and amino groups on free or bound amino acids. Recently, hydrogen sulfide (H(2)S) has been identified as an endogenous vascular gasotransmitter and neuromodulator which can reach up to 160 micromol/l in the brain. Markedly higher 4-HNE concentrations were reported in the brain of patients suffering from Alzheimer's disease. Assuming that the low molecular thiol H(2)S may react with 4 HNE, we have tested the ability of H(2)S to counteract the cytotoxic and protein modifying activity of 4-HNE. The results show that H(2)S at physiologically relevant concentrations could effectively protect neuronal cells (SH-SY5Y) from the cytotoxic action of 4-HNE. The HNE-modification of cellular proteins was also inhibited in presence of H(2)S. These data suggest that H(2)S may be an important protective factor against carbonyl stress by inactivating/modulating the action of highly reactive alpha,beta-unsaturated aldehydes like 4-HNE in the brain. PMID- 19680737 TI - Effects of proNGF on neuronal viability, neurite growth and amyloid-beta metabolism. AB - Alzheimer's disease (AD) is characterized pathologically by the deposition of amyloid-beta peptides (Abeta), neurofibrillary tangles, distinctive neuronal loss and neurite dystrophy. Nerve growth factor (NGF) has been suggested to be involved in the pathogenesis of AD, however, the role of its precursor (proNGF) in AD remains unknown. In this study, we investigated the effect of proNGF on neuron death, neurite growth and Abeta production, in vitro and in vivo. We found that proNGF promotes the death of different cell lines and primary neurons in culture, likely dependent on the expression of p75(NTR). We for the first time found that proNGF has an opposite role in neurite growth to that of mature NGF, retarding neurite growth in both cell lines and primary neurons. proNGF is localized to the Abeta plaques in AD mice brain, however, it had no significant effect on Abeta production in vitro and in vivo. Our findings suggest that proNGF is an important factor involving AD pathogenesis. PMID- 19680738 TI - Simple and rapid analysis of aristolochic acid contained in crude drugs and Kampo formulations with solid-phase extraction and HPLC photodiode-array detection. AB - Simple and rapid analysis of aristolochic acid (AA) in crude drugs and Kampo extracts using a solid-phase extraction method and HPLC-PDA analysis was investigated. Extraction of AA from samples was accomplished by adding methanol containing 1% ammonia. The addition of ammonia ionized the AA of acidic substances so that they adhered to an acrylamide copolymer of a strong anion exchange resin (Sep-Pak QMA) coupled to diol silica easily. Furthermore, a mixture of acetonitrile-water-phosphoric acid (75:25:2, v/v) was effective in isolating AA from its carrier. Since almost all interfering peaks originating from contaminants in crude drugs and Kampo extract formulations could be removed, a satisfactory HPLC chromatogram of AA was obtained. A good result was also obtained when Aristolochiaceae and crude drugs containing AA were tested. Particularly in the case of the medicinal parts of Asarum, several interfering peaks and a ghost peak detected near the AA peak were eliminated. The AA contents of two Kampo extract formulations, tokishigyakukagoshuyushokyoto and ryutanshakanto, were calculated by HPLC analysis. The AA content (the sum of AA-I and AA-II) was 1.25-6.13 mg per daily dose. From an additional recovery experiment for Kampo formulations, high recovery rates of AA were obtained. Neither LC/MS nor special instrumentation was necessary. Our results suggest that this simple, quick, and sensitive analytical method to detect AA in crude drugs and Kampo extract formulations would be valuable in safety inspections of AA in crude drugs and their products. PMID- 19680739 TI - Development of an ultra-high resolution SPECT system with a CdTe semiconductor detector. AB - OBJECTIVE: The aim of this work was to evaluate an ultra-high spatial resolution SPECT system with a semiconductor detector and a high-resolution parallel-hole collimator or a pinhole collimator for small animal imaging. METHODS: We evaluated an ultra-high spatial resolution SPECT system with a high-resolution parallel-hole collimator attached to a cadmium telluride (CdTe) semiconductor detector for small animal imaging. The sizes of an active area and a pixel in the semiconductor detector were 44 x 44 and 0.5 x 0.5 mm(2), respectively. In the high-resolution parallel-hole collimator the size of a hole was 0.4 x 0.4 mm(2), the thickness of a septum 0.1 mm, and the hole-length 30 mm. We also used a high resolution pinhole collimator with a hole size of 0.3 or 0.5 mmvarphi. The physical performance of this SPECT system was evaluated with some experiments with phantoms filled with (99m)Tc-pertechnatate solution. In addition ideal performance and limitations of the system were evaluated with Monte Carlo simulations under the same geometrical conditions as in the experiments. In the evaluation for small animal imaging, we used mice that were administered with (99m)Tc-MDP. We also conducted an ultra-high resolution X-ray CT of the mice to verify the accumulated location of (99m)Tc-MDP using the bone CT images of the mice. RESULTS: The results of the phantom experiments showed that we could resolve 1 mmvarphi hot-channels and 1.6 mmvarphi cold-rods with the high resolution parallel-hole collimator and pinhole collimators. We could image 0.3 mmvarphi hot-channels with the high-resolution pinhole collimators. The results of the simulations showed that the resolution limit in the pinhole imaging was about 0.6 mm FWHM. And the results of experiments with mice showed that we could reconstruct high-resolution images of (99m)Tc-MDP. Furthermore, the distribution of (99m)Tc-MDP in a mouse was found to correspond closely to the location of the bones of the mouse in reconstructions made with the ultra-high resolution X-ray CT system. CONCLUSIONS: Our results demonstrated that the ultra-high spatial resolution SPECT system was feasible for small animal imaging allowing a relatively long data acquisition time. PMID- 19680740 TI - Diagnostic usefulness of 18F-FDG PET/CT in the differentiation of pulmonary artery sarcoma and pulmonary embolism. AB - OBJECTIVE: The purpose of this study was to evaluate the usefulness of 18F fluorodeoxyglucose (FDG) positron emission tomography (PET) in differentiating pulmonary artery sarcoma from pulmonary embolism. MATERIALS AND METHODS: We evaluated three patients with pulmonary artery sarcoma and 10 patients with proximal pulmonary embolism (6 men and 7 women, ranging in age from 35 to 94 years). All the patients had evidence of perfusion defects in their proximal pulmonary arteries on contrast-enhanced computed tomography (CT) scans performed prior to PET/CT. The maximum standardized uptake value (SUV(max)) of FDG uptake in all the lesions was measured using PET/CT. The location of lesions, background uptake or thrombi in the legs were evaluated in both groups as basic characteristics. RESULTS: The mean SUV(max) of the pulmonary artery sarcomas (7.63 +/- 2.21, n = 3) and the pulmonary embolisms (2.31 +/- 0.41, n = 10) were significantly different (P < 0.05). The mean times between the initial contrast enhanced CT scan and PET/CT scan were similar in both groups (P = 0.7804). The differences in the locations in lesions between the three groups (right, left and bilateral) or background uptakes were not significant. CONCLUSION: FDG PET/CT could distinguish pulmonary artery sarcoma from pulmonary embolism based on the SUV(max) value. PMID- 19680742 TI - A radiological investigation of the hepatorenal space: the importance of the ventral decubitus for safer percutaneous right renal access. AB - We investigated anatomical and radiological morphometric parameters, aiming to minimize the risk of hepatic and colonic injuries during right percutaneous kidney access under either ventral or dorsal decubitus of patients. Prone and supine abdominal computerized tomographic examinations from 31 normal adult volunteers (men = 12; women = 19; without history of abdominal pathology) were analyzed morphometrically in order to study the dynamic anatomical relations between the liver and the right kidney. The age of the volunteers ranged from 22 to 64 years old (mean +/- SD = 42.77 +/- 2.10). We observed a significantly greater distance between the liver and the right kidney (hepatorenal space) when the examinee is positioned in ventral decubitus (3.93 +/- 0.37 cm) in comparison with dorsal decubitus (1.98 +/- 0.20 cm). Accordingly, we conclude that right percutaneous access to the inferior right renal pole implies a significantly lower risk (P < 0.01) of both hepatic and biliary injuries when performed in ventral decubitus, comparatively to dorsal decubitus. PMID- 19680741 TI - Immunohistochemical expression of retinoblastoma and phospho-retinoblastoma protein in sheep lung exposed to fluoro-edenite fibers. AB - The immunohistochemical expression of retinoblastoma protein (Rb) and its phosphorylated form (pRb) was investigated in an in vivo sheep model, i.e., lung tissue exposed to asbestos-like fibers. Rb is a phosphoprotein with a central role in controlling the cell cycle, cell growth regulation, differentiation, and apoptosis. pRb functions as a checkpoint in the G(1) phase, favoring cell growth, while the hypophosphorylated form (Rb) not only promotes cell-cycle arrest and a return to the G(1) phase, but also appears to be a key player in most instances of apoptosis. pRb overexpression was detected in alveolar epithelium and the interstitium, especially close to the fluoro-edenite fibers, whereas Rb immunostaining was faint or absent. The data suggest that the altered balance between pRb and Rb expression may be a programmed response to protect the organism against uncontrolled cell proliferation. PMID- 19680743 TI - NT-3 gene modified Schwann cells promote TrkC gene modified mesenchymal stem cells to differentiate into neuron-like cells in vitro. AB - Reports of neuronal differentiation of bone marrow derived mesenchymal stem cells (MSCs) suggested the possibility that these cells could serve as a source of treatment for spinal cord injury. However, the percentages of neuron-like cells differentiated from the MSCs were relatively low both in vitro and in vivo. Here, we investigated whether co-culture of human neurotrophin-3 (NT-3) gene modified Schwann cells (SCs) and human NT-3 receptor tyrosine protein kinase C (TrkC) gene modified MSCs could increase differentiation of neuron-like cells from MSCs. It was shown that MSCs were significantly promoted to differentiate into neuron-like cells, as evidenced immunocytochemically by the expression of neuronal markers, including nestin, beta-III-tubulin, MAP2 and PSD95, 7 days after co-culture. However, the expression of glial fibrillary acidic protein (GFAP)--an astrocyte marker in these cells--was not so obvious. These results demonstrate that the binding of overexpressed NT-3 in SCs and its receptor TrkC in MSCs can be considered to stimulate the increased rate of neuronal differentiation. PMID- 19680744 TI - Middle thymothyroid artery arising from the common carotid artery: case report of a rare variation. AB - A middle thymothyroid artery, arising from the anterior aspect of the right common carotid as an anomalous branch was observed in a 71-year-old Japanese male cadaver. It soon divided into a thyroidea ima artery, a branch supplying the sternoclavicular joints, and a thymic branch. In addition, twigs from these three main branches supplied the sternohyoid and sternothyroid muscles, right inferior parathyroid gland and also some deep cervical lymph nodes. Anatomical features, clinical implications and a brief account of the developmental aspects of this rare variation are included in this report. PMID- 19680745 TI - Molecular characterization of a member of a new species of grapevine viroid. PMID- 19680746 TI - Celiac disease IgA modulates vascular permeability in vitro through the activity of transglutaminase 2 and RhoA. AB - Celiac disease is characterized by the presence of specific autoantibodies targeted against transglutaminase 2 (TG2) in untreated patients' serum and at their production site in the small-bowel mucosa below the basement membrane and around the blood vessels. As these autoantibodies have biological activity in vitro, such as inhibition of angiogenesis, we studied if they might also modulate the endothelial barrier function. Our results show that celiac disease patient autoantibodies increase endothelial permeability for macromolecules, and enhance the binding of lymphocytes to the endothelium and their transendothelial migration when compared to control antibodies in an endothelial cell-based in vitro model. We also demonstrate that these effects are mediated by increased activities of TG2 and RhoA. Since the small bowel mucosal endothelium serves as a "gatekeeper" in inflammatory processes, the disease-specific autoantibodies targeted against TG2 could thus contribute to the pathogenic cascade of celiac disease by increasing blood vessel permeability. PMID- 19680747 TI - Expression of chlorophyll synthase is also involved in feedback-control of chlorophyll biosynthesis. AB - At the last step of the chlorophyll biosynthetic pathway chlorophyll synthase (CHLG) esterifies chlorophyllide a and b with phytyl or geranyl-geranyl pyrophosphate in chloroplasts. Transgenic tobacco plants expressing CHLG RNA in sense and antisense orientation were examined for the effects of excessive and reduced ectopic CHLG expression, respectively, on the chlorophyll biosynthetic pathway and the expression of chlorophyll-binding proteins. Reduced chlorophyll synthase activity does not result in accumulation of chlorophyllide and caused reduced ALA formation and Mg and ferrochelatase activity, while CHLG overexpression correlated with enhanced ALA synthesizing capacity and more chelatase activities. The transcript levels of genes expressing proteins of chlorophyll biosynthesis and chlorophyll-binding proteins were down-regulated in response to reduced CHLG expression. Thus, reduced expression and activity of chlorophyll synthase caused a feedback-controlled inactivation of the initial and rate limiting step of the pathway leading to down regulation of the metabolic flow, while overexpression can mediate a stimulation of the pathway. Chlorophyll synthase is proposed to be important for the co-regulation of the entire pathway and the coordination of synthesis of chlorophyll and the chlorophyll-binding proteins. PMID- 19680748 TI - Population genetic structure of wild and farmed rusa deer (Cervus timorensis russa) in New-Caledonia inferred from polymorphic microsatellite loci. AB - Historical records indicate that 12 rusa deer (Cervus timorensis russa) were introduced in New-Caledonia during the 1870s. We used eight polymorphic microsatellite DNA loci to assess the genetic differentiation and diversity of farmed and wild deer populations. Past genetic bottlenecks were detected in both sub-populations, although higher genetic diversity was maintained in farmed populations, probably due to the regular introduction of reproducers from wild populations and from other farms. The genetic structure of farmed and wild populations differed significantly. There was a significant isolation by distance for wild populations, whereas farmed populations were significantly differentiated between farms independently from their geographical proximity. Wild rusa deer consisted of small populations (with effective population sizes ranging between 7 and 19 individuals depending on the methods used), with a low parent-offspring dispersion range (0.20-2.02 km). Genetic tools and direct observations provided congruent estimates of dispersion and population sizes. We discuss the relevance of our results for management purposes. PMID- 19680749 TI - Is the incidence of invasive vulvar cancer increasing in the United States? AB - OBJECTIVE: To document incidence rates of vulvar cancer, specifically invasive vulvar cancer, from 1973 to 2004 in the United States. METHODS: Nine US cancer registries from the Surveillance, Epidemiology, and End Results (SEER) databases were used to identify women aged 15-84 years, who were first diagnosed with vulvar cancer during 1973-2004. Age-adjusted incidence rates and annual percentage changes were calculated for different time periods, stage of the disease, age, race, and geographic area. RESULTS: During 1973-2004, the incidence of in situ vulvar tumors increased by an average of 3.5% per year (95% CI: 2.9%, 4.1%), while the incidence of invasive tumors increased 1.0% per year (95% CI: 0.6%, 1.4%). An increasing incidence was observed for localized and regional invasive tumors. To at least some degree, the rise of incidence rates of incidence tumors was evident in every age category, race, and geographic region. CONCLUSIONS: Incidence rates of invasive vulvar cancer have increased in the United States during the last three decades. The reasons for this increase are unknown. PMID- 19680750 TI - Polymorphisms in arsenic metabolism genes, urinary arsenic methylation profile and cancer. AB - Arsenic-metabolism-related genes can regulate the arsenic methylation process and may influence susceptibility to cancer. We evaluated the roles of arsenic metabolism genes on urinary arsenic profiles of repeated measurement with 15-year follow-up (1988-2004) through general linear model (GLM) and assessed the effect of the changed extent of urinary arsenic profiles on cancer risk. Questionnaire information and blood samples and two urines (1988 and 2004) were collected from 208 subjects in an arseniasis hyperendemic area in Taiwan. Profiles for concentrations of urinary arsenic were determined using HPLC-HG-AAS. The relative proportion of each arsenic species was calculated by dividing the concentration of each arsenic species by the total arsenic concentration. Genotyping was done using the 5' nuclease allelic discrimination (Taqman) assay. The incidence of cancer was identified through linking to the National Cancer Registry Systems. The Cox proportional hazards model and survival curves were used in the analyses. After a 15-year follow-up, baseline monomethylarsonic acid percentage (MMA%) and change in MMA% exhibited a significant dose-response relationship with cancer risk. Individuals with a higher baseline MMA% and a lower change in MMA% had the earliest cancer incidence (statistically significant). Through GLM, significant gene effects of arsenic (+3 oxidation state)-methyltransferase (AS3MT) on MMA%, dimethylarsinic acid percentage (DMA%) and DMA/MMA, purine nucleoside phosphorylase (PNP) on DMA% and glutathione S-transferase omega 2 (GSTO2) on inorganic arsenics (InAs%) were found. Our results show that MMA% might be a potential predictor of cancer risk. The change in MMA% was linked to individual cancer susceptibility related to AS3MT rs3740393. PMID- 19680751 TI - Metallic components of traffic-induced urban aerosol, their spatial variation, and source apportionment. AB - This study proposes a practical method to estimate elemental composition and distribution in order to attribute source and quantify impacts of aerosol particles at an urban region in Kolkata, India. Twelve-hour total particulates were collected in winter (2005-2006) and analyzed by energy-dispersive X-ray fluorescence technique to determine multi-elemental composition, especially trace metals. The aerosols consist of various elements including K, Ca, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Se, and Pb which exhibit significant concentration at various sites (p < 0.05). The concentration of different metallic elements were found in the order of Zn > Pb > Ni > Cu > Cr > Co. Statistical multivariate analysis and correlation matrix analyses were performed for factor identification and consequent source apportionment. Contour profiles demonstrate spatial variation of elemental compositions indicating possible source contribution along with meteorological influences. Spatial differences were clearly most significant for Zn, Ni, Pb, and Cu reflecting the importance of anthropogenic inputs, primarily from automobile sources. PMID- 19680752 TI - Trihalomethane formation potential in treated water supplies in urban metro city. AB - Trihalomethane (THM) formation potential (TFP) is very useful test to assess the level of the formation of trihalomethanes in worst case scenario. Organics in water have the potential to generate harmful disinfection by-products (DBPs) such as THMs, as a result of their reaction with disinfectant chlorine used in drinking water. DBPs are increasingly recognized as cancerous agents. TFP of postchlorinated treated water were investigated at six water treatment plants (WTPs) in Delhi City (India). The present paper presents the current trends of TFP so that prevention and control measures can be initiated by the regulating agencies responsible for drinking water supply. Liquid-liquid extraction method, followed by qualitative and quantitative estimation by gas chromatograph equipped with electron capture detector, had been used for the determination of THMs in water samples collected at the outlet just before supplying to the consumers during 2000-2007. The TFP values from 2004 onward of all WTPs did not exceed the WHO guideline value of 0.05) of heat stress measurement on days open or services per conception in either multiparous or primiparous cows. During hotter months of calving, multiparous cows showed higher services per conception and primiparous cows showed higher days open and services per conception (P < 0.05). Maximum prepartum degree-days were positively associated (P < 0.05) with calving difficulty score. Multiparous cows with high body condition score at calving were 1.47 times more likely to present a very difficult calving than cows that calved in October (P < 0.05). Collectively, these results suggest that reproductive performance was not affected by cumulative prepartum heat stress although it was associated with very difficult calving score. PMID- 19680775 TI - Serodiagnosis of bovine leptospirosis by IgG-enzyme-linked immunosorbent assay and latex agglutination test. AB - The efficacy of a recombinant leptospiral outer membrane protein LipL41 as an antigen for conducting IgG-Enzyme linked immunosorbent assay (ELISA) and latex agglutination test (LAT) for serodiagnosis of bovine leptospirosis was evaluated. The recombinant LipL41 antigen developed and used for detecting the antibodies was specific in detection of the pathogenic serovars of Leptospira, as the expression of the LipL41 antigen is restricted only to pathogenic leptospires. A total of 430 bovine serum samples were subjected to IgG-ELISA and LAT, and the sensitivity and specificity were assessed in comparison with microscopic agglutination test (MAT). The sensitivity and specificity of IgG-ELISA and LAT were 86.84% and 93.16%, and 95.42% and 98.33% respectively. Both the tests are found to be sensitive, specific and concurred with the standard MAT. The study concluded that the rLipL41 protein could be used as a potential diagnostic antigen in different assay formats for bovine leptospirosis. PMID- 19680776 TI - Erectile dysfunction: a warning sign of silent vascular disease. PMID- 19680778 TI - Purification and characterization of cytochrome c(6) from Acaryochloris marina. AB - Cytochrome c(6), (cyt c(6)) a soluble monoheme electron transport protein, was isolated and characterized from the chlorophyll d-containing cyanobacterium Acaryochoris marina, the type strain MBIC11017. The protein was purified using ammonium sulfate precipitation, ion exchange and gel filtration column chromatography, and fast performance liquid chromatography. Its molecular mass and pI have been determined to be 8.87 kDa and less than 4.2, respectively, by mass spectrometry and isoelectrofocusing (IEF). The protein has an alpha helical structure as indicated by CD (circular dichroism) spectroscopy and a reduction midpoint potential (E(m)) of +327 mV versus the normal hydrogen electrode (NHE) as determined by redox potentiometry. Its potential role in electron transfer processes is discussed. PMID- 19680779 TI - Effect of iron overload on exercise capacity in thalassemic patients with heart failure. AB - In b-thalassemia, myocardial iron overload contributes to heart failure, despite chelation treatment. We hypothesized that myocardial T2*, an index of iron overload, influences patients' physical activity. We assessed a thalassemic population by both cardiovascular magnetic resonance imaging (CMR) and ergospirometry test. Sixty-six thalassemic patients aged 27 (19-40) years, 30 without (NHF) and 36 with heart failure (HF), were studied. Cardiac T2* and left ventricular ejection fraction (LVEF) were evaluated using a 1.5 T system. VO(2max), AT, Mets and duration of exercise by ergospirometry were also assessed. Myocardial T2* was lower in HF compared to NHF patients (14.7 +/- 6.6 vs. 39 +/- 2 ms, P < 0.001). LVEDV and LVESV were higher in HF group compared to NHF patients (139.9 +/- 16.3 vs. 124.6 +/- 20.86 ml, P < 0.01 and 94.9 +/- 24 vs. 38.3 +/- 10.1 ml, P < 0.001, respectively). Additionally, LVEF in HF was lower compared to NHF patients (21.3 +/- 6.1% vs. 69.6 +/- 3.7, P < 0.001, respectively). All exercise parameters were lower in HF compared to NHF patients (P < 0.001). Patients within the HF group were additionally analyzed according to T2* values (<10 ms). HF patients with T2* < 10 ms (n = 13) were considered as high iron overloaded (HF-H) and the rest of them (n = 23) as (HF-L). Although LVEDV, LVESV, LVEF were similar in the two subgroups, the exercise parameters were significantly lower in the HF-H group (P < 0.001). Heart T2* correlated with all exercise parameters (P < 0.001). HF thalassemic patients have reduced exercise indexes compared to non HF. Myocardial iron overload, expressed as T2*, has a direct influence on exercise capacity, independent of LV ejection fraction and functional class. PMID- 19680780 TI - An appearance of "non-compaction" of the right systemic ventricle is common in congenitally corrected transposition of the great arteries. PMID- 19680782 TI - Cloning and characterization of a SnRK1-encoding gene from Malus hupehensis Rehd. and heterologous expression in tomato. AB - Sucrose non-fermenting-1-related protein kinase-1 (SnRK1) plays an important role in metabolic regulation in plant. To understand the molecular mechanism of amino acids and carbohydrate metabolism in Malus hupehensis Rehd. var. pinyiensis Jiang (Pingyi Tiancha, PYTC), a full-length cDNA clone encoding homologue of SnRK1 was isolated from PYTC by Rapid Amplification of cDNA Ends (RACE). The clone, designated as MhSnRK1, contains 2063 nucleotides with an open reading frame of 1548 nucleotides. The deduced 515 amino acids showed high identities with other plant SnRK1 genes. Quantitative real-time PCR analysis revealed this gene was expressed in roots, stems and leaves. Exposing seedlings to nitrate caused and initial decrease in expression of the MhSnRK1 gene in roots, leaves and stems in short term. Ectopic expression of MhSnRK1 in tomato mainly resulted in higher starch content in leaf and red-ripening fruit than wild-type plants. This result supports the hypothesis that overexpression of SnRK1 causes the accumulation of starch in plant cells. All the results suggest that MhSnRK1 may play important roles in carbohydrate and amino acid metabolisms. PMID- 19680781 TI - Comparison of genotypes between environmental and clinical isolates of Cryptococcus neoformans var. grubii based on microsatellite patterns. AB - We applied multilocus microsatellite typing (MLMT) method to investigate the genetic relation between Cryptococcus neoformans var. grubii clinical and environmental isolates in Sao Paulo, Brazil. This MLMT method includes three functional gene sequences of C. neoformans var. grubii, which are dispersed on three chromosomes. In all, 89 strains (36 clinical and 53 environmental isolates) were analyzed. Of 36 clinical strains, 20 belonged to a major type of MLMT-13 (55.6%). They were mainly isolated from clinical specimens. About 52.8% of strains from the environment belong to a major type of MLMT-36, which are indigenous to environments and which were not isolated from clinical samples. Thus, we recognized two genotypes that distinguish majority of clinical and environmental strains. No differences were found in antifungal susceptibility and capsule size between major environmental and clinical MLMT types. PMID- 19680783 TI - IGF-1 gene polymorphism in obese patients with insulin resistance. AB - IGFs (Insulin like growth factors) are important regulators of pancreatic beta cell development, growth and maintenance. Mutations in the IGF genes have been found to be associated with diabetes mellitus, myocardial infarction obesity. These associations could result from changes in insulin secretion. We aimed to investigate IGF-1 gene polymorphism in obese patients with insulin resistance. We included 100 obese patients with insulin resistance 30 healthy subjects to study. At baseline examinations, antropometric measurements were done. Genomic DNA from the patients and controls were prepared. Thyroid function tests and serum IGFBP3 levels were similar between patients and controls whereas IGF, GH levels were significantly lower in obese patients. We categorized the IGF-1 (CA)19 polymorphism area into 3 groups as lower than 192 bp (group 1), 192-194 bp (group 2), and higher than 194 bp(group 3). Group 3 was more frequent in both obese and control groups. IGF-1 levels were also significantly lower in obese group (138.51 +/- 49.3) in than controls (218.14 +/- 69.15). IGF-1 levels were significantly lower in obese patients. The most frequent IGF-1 gen polymorphism allel is >194 bp in both obese insulin resistant patients and controls. IGF-1 levels and the other biochemical and hormonal parameters were similar in different genotype groups. The cause of lower IGF-1 levels in obese patients might be different from IGF-1 gene polymorphism and it may be insulin resistance. PMID- 19680784 TI - Lipopolysaccharide promotes adhesion and invasion of hepatoma cell lines HepG2 and HepG2.2.15. AB - Inflammation and infection have been linked to the bionomics of many cancers, including hepatocellular carcinoma (HCC). Some bacteria, such as Helicobacter pylori, have been found in pathological specimens from patients with HCC. However, little is known about the direct effects of bacteria or their components on hepatoma cells. We analyzed the in vitro proliferation, adhesion and invasion responses of the tumor cell lines HepG2 and HepG2.2.15 to lipopolysaccharide (LPS), a cell wall constituent of Gram-negative bacteria. HepG2 and HepG2.2.15 cells show increased proliferation in response to LPS. Their invasion and adhesion abilities were also increased in response to LPS, which may be related to increased gene expression of interleukin-8 and transforming growth factor beta1. We infer that bacteria may be ignored by immune systems and directly promote adhesion and invasion of hepatoma cells through LPS. PMID- 19680785 TI - Isolation and expression analysis of a novel major latex-like protein (MLP151) gene from Panax ginseng. AB - This is the first reports on isolation and expression analysis of a major latex like protein (MLP151) gene in Panax ginseng C.A. Meyer. A full-length cDNA of MLP151 was 850 bp and contained a 456 bp open reading frame encoding a polypeptide of 151 amino acids. A theoretical pI value of MLP151 was 4.86 and calculated molecular weight was about 16.87 kDa. The MLP homolog proteins are found in various plants and the neighbor-joining analysis revealed that MLP151 has the closest distance with Sn-1 (bell pepper, MLP homolog gene). We analyzed the expression of MLP151 in different levels in various organs of ginseng and plantlet. In the result, the gene was low expressed in plantlet. We treated the ginseng plantlets with nine kinds of different stresses and analyzed the expression profile of MLP151. Transcript levels were significantly induced by stress treatment of light and mannitol, whereas transcript levels were drastically decreased in dark, H(2)O(2), salicylic acid and wounding samples. PMID- 19680786 TI - Characterization of reference genes for quantitative real-time PCR analysis in various tissues of Salvia miltiorrhiza. AB - Five reference genes, 18S, EF1alpha, alpha-Tubulin, Ubiquitin and Actin, from Salvia miltiorrhiza were analyzed as internal controls for gene expression profiling assay using quantitative real-time polymerase chain reaction (qRT-PCR). The five candidate genes were measured for their transcriptional level in seven tissues, including roots, stems, leaves, sepals, petals, stamens and pistils. Then they were ranked by the GeNorm tool. The results showed that Actin and Ubiquitin were the most stable whereas EF1alpha and 18S did not favor normalization of qRT-PCR results in these tissues. Expression levels of the SmDXR gene were studied in parallel, with Actin and Ubiquitin both or each as reference in the seven tissues, and varying relative quantifications of the SmDXR gene in seven tissues. This study indicated that selection of the most stable genes plays an important role in gene expression profiling assays. PMID- 19680788 TI - Isolation of high quality RNA and construction of a suppression subtractive hybridization library from ramie (Boehmeria nivea L. Gaud.). AB - Isolation of high quality RNA from ramie (Boehmeria nivea L. Gaud.) is difficult due to its high levels of polyphenols, polysaccharides, pectin, fat, wax and other secondary metabolites. A modified procedure based on guanidinium isothiocyanate for RNA preparation of ramie was developed in this study. High concentrations (5%, v/v) of guanidinium isothiocyanate, PVP-4000, sodium citrate and sodium lauryl sarcosinate and beta-mercaptoethanol were used in the extraction buffer, together with a low pH sodium acetate (pH 4.0) added to improve the RNA quality. The average yield was about 400 microg RNAg(-1) fresh leaves. One SSH library which was induced by ramie anthracnose was constructed by utilizing the RNA extracted through the present method. These results showed that our protocol was applicable for RNA isolation from recalcitrant ramie tissues. PMID- 19680787 TI - Overexpression of PIP5KL1 suppresses cell proliferation and migration in human gastric cancer cells. AB - Phosphatidylinositol-4-phosphate 5-kinase-like 1 (PIP5KL1), the forth member of phosphatidylinositol-4-phosphate 5-kinases (PIPKs) type I, acts as a scaffold for localization and activation of PIPKs, which mediates numerous cellular processes. However, the role of PIP5KL1 in the development of human cancer is still lacking. We therefore examined the expression of PIP5KL1 in human normal and cancer tissues by tissue microarrays (TMAs). Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence imaging analysis were used to testify the mRNA and protein levels of PIP5KL1 in human gastric cancer cell line (BGC823). The cell proliferation was investigated with 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay. Both wound healing and transwell migration assay were performed to study the cell migration. The phosphorylation of v-akt murine thymoma viral oncogene homolog 1 (AKT1) was determined by western immunoblot analysis. Immunostaining of gastric cancer tissue microarrays revealed a negative correlation between PIP5KL1 overexpression and gastric cancer in situ. Transient transfection PIP5KL1 induced a significant increase expression at both transcriptional and translational levels and consequent robust inhibition of proliferation (P < 0.05) and migration (P < 0.05) of BGC823 cells. Overexpression of PIP5KL1 markedly inhibited (P < 0.05) serum induced phosphorylation of AKT1. Taken together, these studies indicate a functional negative correlation between elevated levels of PIP5KL1 and the development of human gastric cancer, suggesting that PIP5KL1 overexpression may suppress gastric cancer formation. PMID- 19680790 TI - Bivariate microarray analysis: statistical interpretation of two-channel functional genomics data. AB - Conventional statistical methods for interpreting microarray data require large numbers of replicates in order to provide sufficient levels of sensitivity. We recently described a method for identifying differentially-expressed genes in one channel microarray data 1. Based on the idea that the variance structure of microarray data can itself be a reliable measure of noise, this method allows statistically sound interpretation of as few as two replicates per treatment condition. Unlike the one-channel array, the two-channel platform simultaneously compares gene expression in two RNA samples. This leads to covariation of the measured signals. Hence, by accounting for covariation in the variance model, we can significantly increase the power of the statistical test. We believe that this approach has the potential to overcome limitations of existing methods. We present here a novel approach for the analysis of microarray data that involves modeling the variance structure of paired expression data in the context of a Bayesian framework. We also describe a novel statistical test that can be used to identify differentially-expressed genes. This method, bivariate microarray analysis (BMA), demonstrates dramatically improved sensitivity over existing approaches. We show that with only two array replicates, it is possible to detect gene expression changes that are at best detected with six array replicates by other methods. Further, we show that combining results from BMA with Gene Ontology annotation yields biologically significant results in a ligand-treated macrophage cell system. PMID- 19680789 TI - Improvement of heart function in postinfarct heart failure swine models after hepatocyte growth factor gene transfer: comparison of low-, medium- and high-dose groups. AB - Despite advances in surgical and reperfusion therapy, there is no effective therapy currently exists to prevent the progressive decline in cardiac function following myocardial infarction. Hepatocyte growth factor has potent angiogenic and anti-apoptotic activities. The aim of this study was to investigate the therapeutic effect and dose-effect relationship on postinfarction heart failure with different doses of adenovirus-mediated human hepatocyte growth factor (Ad(5) HGF) transference in swine models. Totally twenty swine were randomly divided into four groups: (a) control group (null- Ad(5), 1 ml); (b) low-dose group (1 x 10(9) Pfu/ml Ad(5)-HGF, 1 ml); (c) medium-dose group (5 x 10(9) Pfu/ml Ad(5)-HGF, 1 ml); (d) high-dose group (1 x 10(10) Pfu/ml Ad(5)-HGF, 1 ml). Four weeks after left anterior descending coronary artery (LAD) ligation, different doses of Ad(5) HGF were transferred in three therapeutic groups via right coronary artery. Four and seven weeks after LAD ligation, gate cardiac perfusion imaging was performed to evaluate cardiac perfusion and left ventricular ejection fraction (LVEF). Seven weeks after surgery, the apoptotic index of cardiocyte was observed by TUNEL, the expression of Bcl-2, Bax, alpha-SMA and Factor VIII in the border zones were evaluated by immunohistochemistry, respectively. Four weeks after myocardial infarction, no significant difference was observed among four groups. Three weeks after Ad(5)-HGF transfer, the improvement of cardiac perfusion and LVEF was obviously observed, especially after 1 x 10(10) Pfu Ad(5)-HGF transfer. TUNEL assay showed that 5 x 10(9) Pfu and 1 x 10(10) Pfu Ad(5)-HGF treatment had a obvious reduction in the apoptotic index compared with the null-Ad(5) group, especially after 1 x 10(10) Pfu Ad(5)-HGF treatment. The expression of Bcl-2 protein was increased and the expression of Bax protein was inhibited in the 5 x 10(9) Pfu and 1 x 10(10) Pfu Ad(5)-HGF groups compared with the null-Ad(5) group. The vessel density of Factor VIII(+) and alpha-SMA(+) was increased in Ad(5)-HGF groups compared with the null-Ad(5) group. There were no significant differences in angiogenesis, reducing apoptosis and ameliorating heart function between the 1 x 10(9) Pfu Ad(5)-HGF group and the null-Ad(5) group. Although no statistical difference was observed between 1 x 10(10) Pfu and 5 x 10(9) Pfu Ad(5)-HGF groups, the cardiac protective effects of 1 x 10(10) Pfu Ad(5)-HGF treatment were greater than 5 x 10(9) Pfu Ad(5)-HGF treatment. Different doses of Ad5-HGF injected via noninfarct-related artery could induce angiogenesis, reduce apoptosis and ameliorate heart function, and the cardiac protective effects of 1 x 10(10) Pfu Ad5-HGF is of most significance. PMID- 19680791 TI - Parent-teen interactions as predictors of depressive symptoms in adolescents with headache. AB - This study investigated parent-adolescent conflict, family functioning, and adolescent autonomy as predictors of depressive symptoms in adolescents with primary headache. Frequent headaches during adolescence can have a negative impact on activity levels and psychological functioning. Depression is particularly prevalent in adolescents with headache but little research has examined the role of parent-teen interactions in predicting depressive symptoms. Thirty adolescents diagnosed with migraine or chronic daily headache completed self-report measures of pain intensity, parent-adolescent conflict, family functioning, and depression. Adolescents and their parents also participated in three videotaped interaction tasks, scored by independent raters to assess adolescent autonomy. Regression models revealed that pain intensity, parent adolescent conflict, and autonomy predicted depressive symptoms. Higher levels of conflict, poorer family functioning and lower levels of autonomy were associated with more depressive symptoms. This study highlights the association between parent-teen interactions and psychological functioning in adolescents with primary headache. Implications for intervention are discussed. PMID- 19680792 TI - Do coping strategies discriminate eating disordered individuals better than eating disorder features? An explorative study on female inpatients with anorexia and bulimia nervosa. AB - The purpose of this explorative research was to examine how the COPE (Coping Orientation to Problem Experienced Inventory), an established instrument for measuring coping styles, and EDI-2 (Eating Disorder Inventory-2), a widely used questionnaire for assessing psychological and behavioural features of eating disorders (ED), discriminate among healthy individuals, inpatients with anorexia nervosa (AN) and inpatients with bulimia nervosa (BN). A discriminant analysis approach was used. Results showed that coping styles such as positive attitude, planning and social support are even more discriminative variables than eating disorder features. Implications for further studies are discussed. PMID- 19680793 TI - Age and disability employment discrimination: occupational rehabilitation implications. AB - INTRODUCTION: As concerns grow that a thinning labor force due to retirement will lead to worker shortages, it becomes critical to support positive employment outcomes of groups who have been underutilized, specifically older workers and workers with disabilities. Better understanding perceived age and disability discrimination and their intersection can help rehabilitation specialists and employers address challenges expected as a result of the evolving workforce. METHODS: Using U.S. Equal Employment Opportunity Commission Integrated Mission System data, we investigate the nature of employment discrimination charges that cite the Americans with Disabilities Act or Age Discrimination in Employment Act individually or jointly. We focus on trends in joint filings over time and across categories of age, types of disabilities, and alleged discriminatory behavior. RESULTS: We find that employment discrimination claims that originate from older or disabled workers are concentrated within a subset of issues that include reasonable accommodation, retaliation, and termination. Age-related disabilities are more frequently referenced in joint cases than in the overall pool of ADA filings, while the psychiatric disorders are less often referenced in joint cases. When examining charges made by those protected under both the ADA and ADEA, results from a logit model indicate that in comparison to charges filed under the ADA alone, jointly-filed ADA/ADEA charges are more likely to be filed by older individuals, by those who perceive discrimination in hiring and termination, and to originate from within the smallest firms. CONCLUSION: In light of these findings, rehabilitation and workplace practices to maximize the hiring and retention of older workers and those with disabilities are discussed. PMID- 19680794 TI - Reliability, validity, and cut scores of the south oaks gambling screen (SOGS) for Chinese. AB - We examined the reliability, validity, and classification accuracy of the South Oaks Gambling Screen (SOGS) when adopted for use in Chinese. The DSM-IV criteria for pathological gambling served as the standard against which the classification accuracy of the SOGS was tested. A total of 283 Chinese adults in the community and 94 Chinese treatment-seeking gamblers were recruited. The internal reliability of the SOGS was satisfactory for the general sample and acceptable for the gambling sample. The SOGS was correlated with the DSM-IV criteria items as well as psychosocial and gambling-related problems. Relative to the DSM-IV criteria, the SOGS tended to overestimate the number of pathological gamblers in both samples. In general, we were relatively confident that individuals were not pathological gamblers if the SOGS scores were between 0 and 4 and were pathological gamblers if the SOGS were between 11 and 20. There was about 50-50 chance of being pathological gamblers if the SOGS scores were between 8 and 10. However, the probability of individuals being pathological gamblers was about 0.30 if the SOGS scores were between 5 and 7. We proposed a SOGS cut score of 8 to screen for probable pathological gambling in Chinese societies. PMID- 19680796 TI - Solvent effects on the spectroscopic properties of styrylquinolinium dyes series. AB - The study on the relationship between the structure and spectroscopic properties of styrylquinolinium dyes were carried out by measuring the electronic visible absorption, steady-state and time-resolved fluorescence spectra of quinoline based hemicyanine dyes. The influence of the solvent on absorption and emission spectra and the solvatochromic properties, observed for both ground and first excited states, for all the dyes were applied for the evaluation of their excited state dipole moments. The ground state dipole moments of dyes under the study were established by applying ab initio calculations. The measured, using solvatochromic methods, excited state dipole moments of tested hemicyanines are in the range from 5.38 to 18.90 D and the change in the dipole moments caused by excitation were found to differ from 1.88 to 6.64 D. It was observed that for all tested dyes the dipole moments of the excited states were higher than those of a ground states. The fluorescence lifetime measurements with picosecond resolution was performed for entire series of hemicyanine dyes possessing different dialkylamino groups attached to the phenyl ring. The average lifetimes of the dye fluorescence, determined from the measured data by multi-order exponential decay curve fitting, were in the range from about 120 to 1200 ps at the fluorescence peak wavelength. The fluorescence lifetime measurements were performed for dyes in ethyl acetate solutions. The time-resolved fluorescence spectra measurements allowed to propose the mechanism of the dyes excited states deactivation. PMID- 19680795 TI - Performance in population models for count data, part II: a new SAEM algorithm. AB - Analysis of count data from clinical trials using mixed effect analysis has recently become widely used. However, algorithms available for the parameter estimation, including LAPLACE and Gaussian quadrature (GQ), are associated with certain limitations, including bias in parameter estimates and the long analysis runtime. The stochastic approximation expectation maximization (SAEM) algorithm has proven to be a very efficient and powerful tool in the analysis of continuous data. The aim of this study was to implement and investigate the performance of a new SAEM algorithm for application to count data. A new SAEM algorithm was implemented in MATLAB for estimation of both, parameters and the Fisher information matrix. Stochastic Monte Carlo simulations followed by re-estimation were performed according to scenarios used in previous studies (part I) to investigate properties of alternative algorithms (Plan et al., 2008, Abstr 1372 [ http://wwwpage-meetingorg/?abstract=1372 ]). A single scenario was used to explore six probability distribution models. For parameter estimation, the relative bias was less than 0.92% and 4.13% for fixed and random effects, for all models studied including ones accounting for over- or under-dispersion. Empirical and estimated relative standard errors were similar, with distance between them being <1.7% for all explored scenarios. The longest CPU time was 95 s for parameter estimation and 56 s for SE estimation. The SAEM algorithm was extended for analysis of count data. It provides accurate estimates of both, parameters and standard errors. The estimation is significantly faster compared to LAPLACE and GQ. The algorithm is implemented in Monolix 3.1, (beta-version available in July 2009). PMID- 19680798 TI - Multiple list learning in adults with autism spectrum disorder: parallels with frontal lobe damage or further evidence of diminished relational processing? AB - To test the effects of providing relational cues at encoding and/or retrieval on multi-trial, multi-list free recall in adults with high-functioning autism spectrum disorder (ASD), 16 adults with ASD and 16 matched typical adults learned a first followed by a second categorised list of 24 words. Category labels were provided at encoding, retrieval, both or not at all. Both groups showed enhanced recall when labels were available during encoding or throughout the task. ASD individuals showed reduced recall of the second list and reduced clustering. Clustering and recall were correlated in both groups, which also showed similar levels of subjective organisation. The findings are discussed in relation to theories of frontal and medial temporal lobe contributions to memory in ASD. PMID- 19680799 TI - Biological validation of self-reported condom use among sex workers in Guinea. AB - Self-reported condom use may be prone to social desirability bias. Our aim was to assess the validity of self-reported condom use in a population of female sex workers using prostate specific antigen (PSA) as a gold standard biomarker of recent unprotected vaginal intercourse. We collected data on 223 sex-workers in Conakry, Guinea in order to assess the sensitivity and specificity of self reported condom use as well as to examine the predictors of discordance between self-report and PSA presence. PSA was detected in 38.4% of samples. Sensitivity of self-reported condom use was 14.6% and its specificity was 94.7%. Self perceived high risk of HIV infection was the only significant independent predictor of misreported condom use. PSA could be useful to validate self reported condom use in surveys and to allow a better understanding of factors associated with social desirability in sexual behaviour reporting. PMID- 19680800 TI - AIDS-related stigma among Black and Hispanic young adults. AB - Telephone surveys with national probability samples of English-speaking adults have suggested that popular support for punitive policies toward people living with HIV/AIDS (PLWHA) declined in the 1990s, but AIDS-related stigma persists in the United States. Our aim was to assess the prevalence and impact of AIDS related stigma in non-Hispanic Black and Hispanic communities. A cross-sectional computer-assisted telephone-interview survey was conducted in summer 2003 with African-American, Afro-Caribbean, Haitian, and Hispanic 18-39 year-old residents of 12 high AIDS-incidence areas in Broward County, Florida. Stigma items were adopted from national surveys, but interviews were conducted in Spanish and Haitian Creole as well as in English. Stigma scores were higher than those reported for national samples, especially among Haitians interviewed in Creole. AIDS-related stigma was associated with never receiving an HIV-antibody test (adjusted odds ratio [AOR] = 0.78, 95% confidence interval [CI]: 0.62-0.99, P = .046), an elevated perception of HIV risk (AOR = 1.32, 95% CI: 1.01-1.73, P = .045) and a failure to participate in HIV-prevention efforts (AOR = 0.53, 95% CI: 0.34-0.85, P = .008). Interventions are needed to mitigate the pernicious effects of AIDS-related stigma. PMID- 19680801 TI - Fish based preimplantation genetic diagnosis to prevent DiGeorge syndrome. AB - PURPOSE: To report the performance of fluorescence in-situ hybridization in the setting of preimplantation genetic diagnosis in order to diagnose embryos affected by DiGeorge syndrome. DESIGN: Case report. SETTING: Academic referral center. PATIENT: A 32 year-old female affected by DiGeorge syndrome. INTERVENTION(S): History and physical examination, karyotyping, amniocentesis, preimplantation genetic diagnosis, fluorescence in-situ hybridization. MAIN OUTCOME MEASURE(S): Avoidance of pregnancy with embryo affected by DiGeorge syndrome. RESULT(S): Termination of pregnancy with an affected embryo followed by fluorescence in-situ hybridization based preimplantation genetic diagnosis and delivery of healthy offspring. CONCLUSION(S): The combination of preimplantation genetic diagnosis with fluorescence in-situ hybridization is recommended to prevent pregnancies with DiGeorge syndrome affected embryos in properly selected patients. PMID- 19680802 TI - Case report: successful delivery following the transfer of a human re-vitrified day-7 spontaneously hatched blastocyst developed from vitrified cleaved embryos. AB - PURPOSE: To report a successful delivery after the transfer of a re-cryopreserved day-7 hatched blastocyst. METHODS AND RESULTS: A 30-year-old woman underwent a long-treatment protocol for ovarian stimulation. Fourteen mature oocytes were obtained, and twelve were fertilized. On day 3, two cleaved embryos were transferred, but no implantation occurred. The remaining embryos were vitrified. Subsequently, two vitrified day-3 embryos were transferred. The woman became pregnant and delivered a healthy baby. Subsequently, two vitrified day-3 embryos were transferred, but no pregnancy occurred. Subsequently, all the remaining vitrified day-3 embryos were cultured. On day 5, no blastocyst was obtained. The remaining embryos were continued to be cultured. On day 7, a hatched blastocyst was obtained and re-vitrified. Subsequently, the re-vitrified day-7 hatched blastocyst was transferred. The woman became pregnant and delivered a healthy female. CONCLUSIONS: The day-7 hatched blastocyst developed from vitrified embryos can be re-vitrified and have pregnancy potential after re-warming. PMID- 19680803 TI - Congenital anomalies and other perinatal outcomes in ICSI vs. naturally conceived pregnancies: a comparative study. AB - BACKGROUND: Intracytoplasmic sperm injection (ICSI) procedures have become accepted worldwide and their effect on society is well-known. However, the full extent of the possible complications of these procedures on maternal and neonatal outcome is still unclear. MATERIALS AND METHODS: This is a retrospective case controlled study from January 2003 to December 2007 which compared 253 women that had conceived using assisted reproduction (ICSI) and delivered 327 children at our center (study group) with a matched group of 349 women who naturally conceived and delivered 354 children at Abha General Hospital (control group) during the same period. The obstetrical and neonatal characteristics of the women and their children were assessed to determine any significant differences between the groups. RESULTS: The number of gestations per pregnancy (1.34 +/- 0.57 vs. 1.01 +/- 0.12) and number of children born per woman (1.28 +/- 0.49 vs. 1.01 +/- 0.12) was significantly higher in the ICSI group (p < 0.001). In addition, the gestational age at delivery (37.23 +/- 2.68 vs. 38.56 +/- 1.89) was significantly shorter in the ICSI group (p < 0.001) and this led to an increased number of obstetrical interventions, as well as the incidence of cesarean deliveries. Examination of the new-born children revealed similar incidence of congenital anomalies in both groups. CONCLUSION: ICSI conceived pregnancies were characterized by an increased number of gestations and live-born, and there was no increase in congenital malformations compared to naturally conceived pregnancies. PMID- 19680804 TI - Peer perceptions of social skills in socially anxious and nonanxious adolescents. AB - Previous studies using adult observers are inconsistent with regard to social skills deficits in nonclinical socially anxious youth. The present study investigated whether same age peers perceive a lack of social skills in the socially anxious. Twenty high and 20 low socially anxious adolescents (13-17 years old) were recorded giving a 5-min speech. Unfamiliar peer observers (12-17 years old) viewed the speech samples and rated four social skills: speech content, facial expressions, posture and body movement, and way of speaking. Peer observers perceived high socially anxious adolescents as significantly poorer than low socially anxious adolescents on all four social skills. Moreover, for all skills except facial expressions, group differences could not be attributed to adolescents' self-reported level of depression. We suggest that therapists take the perceptions of same age peers into account when assessing the social skills of socially anxious youth. PMID- 19680805 TI - Pre-sleep arousal and sleep problems of anxiety-disordered youth. AB - The current study examined sleep problems and pre-sleep arousal among 52 anxious children and adolescents, aged 7-14 years, in relation to age, sex, ethnicity, and primary anxiety disorder. Assessment included structured diagnostic interviews and parent and child completed measures of sleep problems and pre sleep arousal. Overall, 85% of parents reported clinically-significant child sleep problems, whereas 54% of youth reported trouble sleeping. Young children, those with primary generalized anxiety disorder, and Latino youth experienced the greatest levels of sleep disturbance. Additionally, greater levels of pre-sleep cognitive rather than somatic arousal were found and pre-sleep thoughts were associated with decreased total sleep duration and greater sleep problems. Findings suggest that attention to sleep should be part of assessment procedures for anxious children in both research and clinical settings. PMID- 19680807 TI - Effects of hydroxysafflor yellow A in the attenuation of MPTP neurotoxicity in mice. AB - Parkinson's disease (PD) is a progressive neurodegenerative disorder whose etiology is not understood. The 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated mouse model is widely used for studying PD. The present study was undertaken to investigate the effect of hydroxysafflor yellow A (HSYA) on MPTP induced neurotoxicity in mice. Pretreatment with HSYA at a dose of 2, 8 mg/kg for a week was followed by intraperitoneal injection with MPTP (30 mg/kg) for five consecutive days. Next, the subsequent behavior, biochemical index and immunohistochemical manifestations in mice were determined. Behavioral testing showed that MPTP-treated mice exhibited motor deficits but HSYA at dose of 8 mg/kg prevented the appearance of motor abnormalities. Treatment with HSYA at dose of 8 mg/kg attenuated the reduction of dopamine (DA), 3,4 dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in striatum. It also showed that the activity of SOD, catalase activity and GSH levels were significantly higher, while the levels of malondialdehyde (MDA) and hydroxyl radicals was lower, in the HSYA-treated mice compared to the MPTP-treated mice. The MPTP-treated mice exhibited the loss of tyrosine hydroxylase-containing dopaminergic neurons in substantia nigra. However, HSYA-treated mice showed a protective effect. Our results indicated that HSYA possesses neuroprotective effects and is a promising anti-Parkinson's disease drug which is worthy of further study. PMID- 19680808 TI - Hepatic stellate cell and myofibroblast-like cell gene expression in the explanted cirrhotic livers of patients undergoing liver transplantation. AB - BACKGROUND: Hepatic stellate cells (HSC) are involved in hepatic fibrogenesis. Cell signaling associated with an insult to the liver affects an HSC transdifferentiation to fibrogenic myofibroblast-like cells. AIMS: To investigate the transcriptional expression distinguishing HSC and myofibroblast-like cells between livers with and without cirrhosis. METHODS: Tissue from ten cirrhotic livers (undergoing transplant) and four non-cirrhotic livers from the National Disease Research Interchange underwent cell separation to extract HSC and myofibroblast-like cell populations. Separated cell types as well as LI-90 cells were subjected to microarray analysis. Selected microarray results were verified by quantitative real-time PCR. RESULTS: Differential expression of some genes, such as IL-1beta, IL-1alpha, and IL-6, was associated with both transdifferentiation and disease. Other genes, such as fatty acid 2-hydroxylase only show differential expression in association with disease. Functional analysis supported these findings, indicating some signal transduction pathways (IL-6) are involved in disease and activation, whereas retinoid X receptor signaling in HSC from cirrhotic and non-cirrhotic livers varies in scope and quality. CONCLUSIONS: These findings indicate distinct phenotypes for HSC from cirrhotic and non-cirrhotic livers. Furthermore, coordinated differential expression between genes involved in the same signal transduction pathways provides some insight into the mechanisms that may control the balance between fibrogenesis and fibrolysis. PMID- 19680809 TI - Activated protein C improves the severity of severe acute pancreatitis via up regulating the expressions of endothelial cell protein C receptor and thrombomodulin. AB - BACKGROUND AND AIMS: Activated protein C (APC) is increasingly understood to have diverse regulatory functions in inflammation. However, the exact mechanism of action remains unclear in severe acute pancreatitis (SAP). The aim of this study was to demonstrate the effects of APC on expressions of thrombomodulin (TM) and endothelial cell protein C receptor (EPCR), and its subsequent effect on the severity of SAP. METHODS: Sprague-Dawley rats were randomly divided into four groups. The rats were given intravenous injections of APC (50, 10 microg/kg, respectively, treated groups) or saline (SAP group) just before induction of SAP. One group of rats underwent only sham operation as control group. Experimental samples were harvested at 16 h after induction. The protein and mRNA levels of matrix metalloprotease 9 (MMP-9), TM, and EPCR in pancreatic tissue were investigated. Serum tumor necrosis factor alpha (TNF-alpha) and interleukin-8 (IL 8) levels were determined. The severity of disease was evaluated by histological score of pancreatic injury, wet/dry weight ratio of pancreatic tissue, and serum amylase. RESULTS: In the APC 50 microg/kg-treated group, serum TNF-alpha, IL-8, and pancreatic MMP-9 levels were decreased and the levels of pancreatic EPCR and TM were up-regulated compared with the SAP group. A significant dose-dependent relationship was found between the decreased levels of serum IL-8 and the APC treated dosage. Furthermore, the severity of SAP was ameliorated by APC treatment. CONCLUSIONS: APC could augment the anti-coagulation and anti inflammatory activity by up-regulating EPCR and TM expressions, thus attenuating the severity of SAP. PMID- 19680810 TI - Immunopathology on hepatocyte expression of HBV surface, core, and x antigens in chronic hepatitis B: clinical and virological correlation. AB - PURPOSE: Hepatocyte expression of HBV surface, core, and x antigens (HBsAg, HBcAg and HBxAg), semi-quantitated by immunopathology, were correlated with clinical and virological data in 80 patients with chronic hepatitis B. RESULTS: Seventy patients were HBsAg positive in cytoplasm, 61 were HBcAg positive, including 45 in both nucleus and cytoplasm and 16 in cytoplasm only, and 47 were HBxAg positive in cytoplasm. The detection rates for HBcAg increased while those for HBsAg and HBxAg decreased with HBV DNA levels. Positive HBcAg staining usually suggested the presence of HBV DNA levels >10(6) copies/ml. HBcAg, HBsAg, and HBxAg expressions showed no significant differences between patients with genotype B and C. Serum HBeAg and HBV DNA levels correlated positively with nuclear or cytoplasmic HBcAg expression but inversely with HBsAg expression. By multiple regression analysis, HBV DNA levels correlated significantly only with nuclear HBcAg expression. ALT levels and inflammatory grades correlated with cytoplasmic HBcAg expression. There was an inverse quantitative relationship between HBcAg and HBsAg expression. Furthermore, HBxAg expression correlated significantly with HBsAg expression as well as male gender. CONCLUSIONS: With diminishing HBV DNA levels following HBeAg seroconversion, HBcAg expression decreased but HBsAg expression increased with a concomitant increase in HBxAg expression. Whether the finding that a significantly higher expression of HBxAg observed in males than females may account for the gender difference in long-term sequelae of chronic HBV infection needs further investigation. PMID- 19680806 TI - Oxidative mechanisms of brain dysfunction during sepsis. AB - Oxidative stress has drawn a lot of attention in the past few decades, since it has been reported to participate in the mechanism of many diseases. Therefore, it seemed to be a good rationale to aim oxidative stress on therapeutic research. Sepsis is a complex systemic syndrome characterized by an imbalance between pro- and anti-inflammatory responses to a pathogen; its pathophysiology is a dynamic process which involves components of the immune system, the coagulation pathway, parenchymal cells, and the endocrine and metabolic pathways. It is well characterized that oxidative stress plays a crucial role in sepsis development, but the relation between central nervous system dysfunction and oxidative stress during sepsis is not well understood. Thus, we here summarize the current knowledge on the role of free radicals in the development of brain dysfunction in sepsis focusing on oxidative damage and the redox control of brain inflammatory pathways. PMID- 19680813 TI - Characterization of cell mechanical properties by computational modeling of parallel plate compression. AB - A substantial body of work has been reported in which the mechanical properties of adherent cells were characterized using compression testing in tandem with computational modeling. However, a number of important issues remain to be addressed. In the current study, using computational analyses, the effect of cell compressibility on the force required to deform spread cells is investigated and the possibility that stiffening of the cell cytoplasm occurs during spreading is examined based on published experimental compression test data. The effect of viscoelasticity on cell compression is considered and difficulties in performing a complete characterization of the viscoelastic properties of a cell nucleus and cytoplasm by this method are highlighted. Finally, a non-linear force-deformation response is simulated using differing linear viscoelastic properties for the cell nucleus and the cell cytoplasm. PMID- 19680811 TI - Are female psychiatry residents better to propose in emergency a voluntary hospitalization? AB - This study analyses assessment, intervention and admission decisions made by emergency psychiatry residents, to determine whether these differ depending on the gender of the resident. Data from all patients presenting to a psychiatric emergency room were collected prospectively for a 3 months period as part of a local quality check project. A questionnaire was used to collect patient demographic data, diagnosis, treatment decisions and the personal and professional characteristics of the residents who performed the assessments. During the 3 months of the study period we obtained data on all 251 emergency assessments carried out by all six residents working in the service. These were 3 female and 3 male 3rd year residents in psychiatry. There was no difference between male and female residents concerning ICD-10 diagnostic assessment, adherence to local hospitalization criteria guidelines, psychotherapeutical and pharmacological treatments administered. A similar distribution between male and female residents was found for diagnoses. No difference was found in the rate of hospitalization decisions between male and female residents. However, surprisingly, there were more voluntary hospitalizations by the women residents (P = 0.035; chi2 = 4.443) and more involuntary admissions by the men residents (P = 0.005; chi2 = 7.643). There was no correlation between the gender of the patient and the assessment or hospitalization decision of either male or female residents. Although this study has methodological limitations, it suggests that female emergency psychiatry residents are more likely to propose voluntary hospitalizations. PMID- 19680812 TI - Experimental validation of a theoretical model of cytokine capture using a hemoadsorption device. AB - Sepsis, a systemic inflammatory response in the presence of an infection, is characterized by overproduction of inflammatory mediators called cytokines. Removal of these cytokines using an extracorporeal hemoadsorption device is a potential therapy for sepsis. We are developing a cytokine adsorption device (CAD) filled with microporous polymer beads and have previously published a mathematical model which predicts the time course of cytokine removal by the device. The goal of this study was to show that the model can experimentally predict the rate of cytokine capture associated with key design and operational parameters of the CAD. We spiked IL-6, IL-10, and TNF into horse serum and perfused it through an appropriately scaled-down CAD and measured the change in concentration of the cytokines over time. These data were fit to the mathematical model to determine a single model parameter, Gamma( i ), which is only a function of the cytokine-polymer interaction and the cytokine effective diffusion coefficient in the porous matrix. We compared Gamma( i ) values, which by definition should not change between experiments. Our results indicate that the Gamma( i ) value for a specific cytokine was statistically independent of all other parameters in the model, including initial cytokine concentration, flow rate, serum reservoir volume, CAD size, and bead size. Our results also indicate that competitive adsorption of cytokines and other middle-molecular weight proteins, which is neglected in the model, does not affect the rate of removal of a given cytokine. The model of cytokine capture in the CAD developed in this study will be integrated with a systems model of sepsis to simulate the progression of sepsis in humans and to develop a therapeutic CAD design and intervention protocol that improves patient outcomes in sepsis. PMID- 19680814 TI - Predicting high school truancy among students in the Appalachian south. AB - Truancy is a considerable problem among adolescents. Considering the historical emphasis on studying truancy in urban regions, a concerted effort is needed to extend this research into rural areas to examine cultural generalizability of findings. The purpose of this study was to assess variables associated with truancy in a rural sample (N = 367) of students attending high school in a southern rural region of the Appalachian Mountains. The primary objective was to assess the relative predictive strength of the following variables: academic performance, religiosity, environmental factors (family structure, parental education, and adolescent perceptions of family functioning), internalizing problems (anxiety, depression, thought problems, attentional problems), externalizing problems (substance use and rule-breaking behaviors), and prosocial overt behaviors (participation in school and leadership activities). Regression analysis indicated that truancy was significantly associated with poor school performance, increased depression, social problems, having a less educated mother, a less structured home environment, higher grade, and decreased participation in school sports. EDITORS' STRATEGIC IMPLICATIONS: These findings are critical for the understanding of truancy in rural areas, and they highlight contextual factors that must be identified and addressed through systematic prevention programs targeting adolescents at risk for truancy. PMID- 19680816 TI - The neuropsychology of schizophrenia circa 2009. PMID- 19680815 TI - Pilot study to gauge acceptability of a mindfulness-based, family-focused preventive intervention. AB - The purpose of the present study was to conduct a test of acceptability of a new model for family-focused drug prevention programs for families of early adolescents. An existing evidence-based behavioral intervention, the Strengthening Families Program: For Parents and Youth 10-14 (SFP), was adapted to include concepts and activities related to mindfulness and mindful parenting (an extension of mindfulness to the interpersonal domain of parent-child relationships). The foundation for this innovative intervention approach stems from research on the effects of mind-body treatments involving mindfulness meditation and the function of stress and coping in relation to parenting and parent well-being. One group of families participated in a seven-week pilot of this mindfulness-enhanced version of SFP. Results of a mixed-method implementation evaluation suggest that the new intervention activities were generally feasible to deliver, acceptable to participants, and perceived to yield positive benefits for family functioning and parent psychological well-being. The next phase of this research will involve curriculum refinement based upon results of this initial study, and a larger pilot efficacy trial will be conducted. PMID- 19680819 TI - Purification and characterization of two extracellular xylanases from Penicillium sclerotiorum: a novel acidophilic xylanase. AB - Two xylanases from the crude culture filtrate of Penicillium sclerotiorum were purified to homogeneity by a rapid and efficient procedure, using ion-exchange and molecular exclusion chromatography. Molecular masses estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were 23.9 and 33.1 kDa for xylanase I and II, respectively. The native enzymes' molecular masses of 23.8 and 30.8 kDa were estimated for xylanase I and II, respectively, by molecular exclusion chromatography. Both enzymes are glycoproteins with optimum temperature and pH of 50 degrees C and pH 2.5 for xylanase I and 55 degrees C and pH 4.5 for xylanase II. The reducing agents beta-mercaptoethanol and dithio-treitol enhanced xylanase activities, while the ions Hg(2+) and Cu(2+) as well the detergent SDS were strong inhibitors of both enzymes, but xylanase II was stimulated when incubated with Mn(2+). The K (m) value of xylanase I for birchwood xylan and for oat spelt xylan were 6.5 and 2.6 mg mL(-1), respectively, whereas the K (m) values of xylanase II for these substrates were 26.61 and 23.45 mg mL(-1). The hydrolysis of oat spelt xylan by xylanase I released xylobiose and larger xylooligosaccharides while xylooligosaccharides with a decreasing polymerization degree up to xylotriose were observed by the action of xylanase II. The present study is among the first works to examine and describe an extracellular, highly acidophilic xylanase, with an unusual optimum pH at 2.5. Previously, only one work described a xylanase with optimum pH 2.0. This novel xylanase showed interesting characteristics for biotechnological process such as feed and food industries. PMID- 19680818 TI - The utilization of gum tragacanth to improve the growth of Rhodotorula aurantiaca and the production of gamma-decalactone in large scale. AB - The production of gamma-decalactone and 4-hydroxydecanoic acid by the psychrophilic yeast R. aurantiaca was studied. The effect of both compounds on the growth of R. aurantiaca was also investigated and our results show that gamma decalactone must be one of the limiting factors for its production. The addition of gum tragacanth to the medium at concentrations of 3 and 4 g/l seems to be an adequate strategy to enhance gamma-decalactone production and to reduce its toxicity towards the cell. The production of gamma-decalactone and 4 hydroxydecanoic acid was significantly higher in 20-l bioreactor than in 100-l bioreactor. By using 20 g/l of castor oil, 6.5 and 4.5 g/l of gamma-decalactone were extracted after acidification at pH 2.0 and distillation at 100 degrees C for 45 min in 20- and 100-l bioreactors, respectively. We propose a process at industrial scale using a psychrophilic yeast to produce naturally gamma decalactone from castor oil which acts also as a detoxifying agent; moreover the process was improved by adding a natural gum. PMID- 19680817 TI - Life and death of microglia. AB - The importance of microglial cells in the maintenance of a well-functioning central nervous system (CNS) cannot be overstated. As descendants of the myelomonocytic lineage they are industrious housekeepers and watchful sentries that safeguard a homeostatic environment through a number of mechanisms designed to provide protection of fastidious neurons at all times. Microglia become particularly active after homeostasis has been perturbed by physical injury or other insults and they enter into a state of activation which is determined largely by the nature and severity of the lesion. Microglial activation is the main cellular event in acute neuroinflammation and essential for wound healing in the CNS. Recent studies from this laboratory have been focused on microglia in the aging brain and identified structural abnormalities, termed microglial dystrophy, that are consistent with cell senescence and progress to a form of accidental cell death that is marked by cytoplasmic degeneration and has been termed cytorrhexis. Cytorrhexis of microglia is infrequent in the normally aged human brain and non-detectable in aged rodents, but its occurrence increases dramatically during neurodegenerative conditions, including Alzheimer's disease (AD) in humans and motoneuron disease in transgenic rats. The identification of degenerating microglia has given rise to a novel theory of AD pathogenesis, the microglial dysfunction hypothesis, which views the loss of microglial neuroprotection as a central event in neurodegenerative disease development. PMID- 19680820 TI - Treatment with tertiary oximes prevents seizures and improves survival following sarin intoxication. AB - The capability of the tertiary oximes, monoisonitrosoacetone (MINA) and diacetylmonoxime (DAM), to reactivate acetylcholinesterase (AChE) inhibited by sarin (GB) in the blood, brain, and peripheral tissues of guinea pigs was compared with that of the quaternary oximes 2-PAM, HLo7, and MMB-4. Animals were injected subcutaneously (s.c.) with 1.0 x LD(50) of GB and treated intramuscularly (i.m.) 5 min later with one of these oximes. Sixty minutes after GB exposure, tissues were collected for AChE analysis. At low doses, MINA and DAM produced significant increases in AChE activity in all brain areas examined, but no significant AChE reactivation in peripheral tissues or blood. At higher doses, MINA and DAM increased AChE activity in the brain, peripheral tissues, and blood. In contrast, the quaternary oximes produced significant reactivation in peripheral tissues and blood AChE, but no significant reactivation of brain AChE. In another study, animals were pretreated i.m. with pyridostigmine 30 min prior to s.c. challenge with 2.0 x LD(50) of GB and treated i.m. 1 min later with atropine sulfate (2.0 mg/kg), plus a varied dose of oximes. MINA and DAM prevented or terminated GB-induced seizure activity and protected against GB lethality in a dose-dependent fashion. In contrast, none of the quaternary oximes prevented or stopped GB-induced seizures. Thus, tertiary oximes reactivated AChE in the brain, improved survival, and terminated seizures following GB intoxication. PMID- 19680821 TI - Expression and Localization of PRiMA-linked globular form acetylcholinesterase in vertebrate neuromuscular junctions. AB - Acetylcholinesterase (AChE) is well known to process different molecular forms via the distinct interacting partners. Proline-rich membrane anchor (PRiMA) linked tetrameric globular AChE (G4 AChE) is mainly found in the vertebrate brain; however, recent studies from our laboratory have suggested its existence at neuromuscular junctions (nmjs). Both muscle and motor neuron express AChE at the nmjs. In muscle, the expression of PRiMA-linked AChE is down-regulated during myogenic differentiation and by motor neuron innervation. As compared with muscle, spinal cord possessed higher total AChE activity and contained PRiMA linked AChE forms. The spinal cord expression of this form increased during development. More importantly, PRiMA-linked G4 AChE identified as aggregates localized at nmjs. These findings suggest that the restricted localization of PRiMA-linked G4 AChE at the nmjs could be contributed by the pre-synaptic motor neuron and/or the post-synaptic muscle fiber. PMID- 19680822 TI - Cholinergic involvement in Alzheimer's disease. A link with NGF maturation and degradation. AB - Basal forebrain cholinergic neurons are highly dependent on nerve growth factor (NGF) supply for the maintenance of their cholinergic phenotype as well as their cholinergic synaptic integrity. The precursor form of NGF, proNGF, abounds in the CNS and is highly elevated in Alzheimer's disease. In order to obtain a deeper understanding of the NGF biology in the CNS, we have performed a series of ex vivo and in vivo investigations to elucidate the mechanisms of release, maturation and degradation of this neurotrophin. In this short review, we describe this NGF metabolic pathway, its significance for the maintenance of basal cholinergic neurons, and its dysregulation in Alzheimer's disease. We are proposing that the conversion of proNGF to mature NGF occurs in the extracellular space by the coordinated action of zymogens, convertases, and endogenous regulators, which are released in the extracellular space in an activity dependent fashion. We further discuss our findings of a diminished conversion of the NGF precursor molecule to its mature form in Alzheimer's disease as well as an augmented degradation of mature NGF. These combined effects on NGF metabolism would explain the well-known cholinergic atrophy found in Alzheimer's disease and would offer new therapeutic opportunities aimed at correcting the NGF dysmetabolism along with Abeta-induced inflammatory responses. PMID- 19680823 TI - Differential regulation of alpha7 nicotinic receptor gene (CHRNA7) expression in schizophrenic smokers. AB - The alpha7 neuronal nicotinic receptor gene (CHRNA7) has been implicated in the pathophysiology of schizophrenia by genetic and pharmacological studies. Expression of the alpha7* receptor, as measured by [(125)I]alpha-bungarotoxin autoradiography, is decreased in postmortem brain of schizophrenic subjects compared to non-mentally ill controls. Most schizophrenic patients are heavy smokers, with high levels of serum cotinine. Smoking changes the expression of multiple genes and differentially regulates gene expression in schizophrenic hippocampus. We examined the effects of smoking on CHRNA7 expression in the same tissue and find that smoking differentially regulates expression of both mRNA and protein for this gene. CHRNA7 mRNA and protein levels are significantly lower in schizophrenic nonsmokers compared to control nonsmokers and are brought to control levels in schizophrenic smokers. Sufficient protein but low surface expression of the alpha7* receptor, seen in the autoradiographic studies, suggests aberrant assembly or trafficking of the receptor. PMID- 19680824 TI - Bedside ultrasound screening for pretracheal vascular structures may minimize the risks of percutaneous dilatational tracheostomy. AB - BACKGROUND AND PURPOSE: Percutaneous dilatational tracheostomy (PDT) continues to gain in popularity as a bedside method for tracheostomy placement in the intensive care unit. Here, we present a description of ultrasound technique and two case examples to show the utility of bedside ultrasound screening to select patients with appropriate anatomy for PDT. METHODS: We have instituted a protocol at our institution to use bedside screening ultrasound to confirm appropriate anatomy prior to PDT. In this report, we present our ultrasound methodology and present two cases with clear correlations between screening ultrasound findings and intraoperative findings. RESULTS: We describe an easily applied method for pretracheal ultrasound screening. To show the utility of this screening technique, we then present two example cases showing pretracheal vascular structures seen on ultrasound and during open operative exploration. CONCLUSION: Bedside ultrasound screening allows for easy identification of pretracheal vascular structures that might pose a hemorrhage risk during PDT. PMID- 19680825 TI - Microarray analysis of altered gene expression in ERbeta-overexpressing HEK293 cells. AB - Estrogen receptors (ERs), ERalpha and ERbeta, mediate estrogen actions in a broad range of target tissues. With the introduction of microarray techniques, a significant understanding has been gained regarding the interplay between the ERalpha and ERbeta in breast cancer cell lines. To gain a more comprehensive understanding of ERbeta-dependent gene regulation independent of ERalpha, we performed microarray analysis on HEK293/mock and HEK293/ERbeta cells. A total of 332 genes was identified as ERbeta-upregulated genes and 210 identified as ERbeta downregulated genes. ERbeta-induced and ERbeta-repressed genes were involved in cell-cell signaling, morphogenesis, and cell proliferation. The ERbeta repressive effect on genes related to proliferation was further studied by proliferation assays, where ERbeta expression resulted in a significant decrease in cell proliferation. To identify primary ERbeta target genes, we examined a number of ERbeta-regulated genes using chromatin immunoprecipitation assays for regions bound by ERbeta. Our results showed that ERbeta recruitment was significant to regions associated with 12 genes (IL1RAP, TMSB4X, COLEC12, ENPP2, KLRC1, RERG, RGS16, TNNT2, CYR61, FER1L3, FAM108A1, and CYP4X1), suggesting that these genes are likely to be ERbeta primary target genes. This study has provided novel information on the gene regulatory function of ERbeta independent of ERalpha and identified a number of ERbeta primary target genes. The results of Gene Ontology analysis and proliferation assays are consistent with an antiproliferative role of ERbeta independent of ERalpha. PMID- 19680826 TI - Intracystic papillary carcinoma of the breast: one of the youngest patient in the literature. PMID- 19680827 TI - Suppression of matrix metalloproteinase-9 expression by RNA interference inhibits SGC7901 gastric adenocarcinoma cell growth and invasion in vitro and in vivo. AB - Matrix metalloproteinase-9 (MMP-9) is considered the key enzyme that degrades extracellular matrix (ECM) via breaking down type IV collagens. Up-regulated MMP 9 promotes growth and invasion of gastric adenocarcinomas. The present study is to block MMP-9 expression in gastric adenocarcinoma cells in order to inhibit tumor growth and invasion. The association between MMP-9 expression and tumor pathology was reconfirmed by applying immunohistochemistry on tissue arrays. Small interference RNAs (siRNA) targeted on human MMP-9 were used to suppress gene expression in SGC7901 human gastric adenocarcinoma cells. Cell growth and invasion were significantly inhibited in specific siRNA-targeted cells. In addition, we generate a SGC7901-subcutaneous mice model to observe anti-tumor effects from RNA interference (RNAi). Data showed tumor masses in MMP-9 siRNA treated mice were significantly smaller than those in control mice. The expression of vascular endothelial growth factor and proliferating cell nuclear antigen were down-regulated in MMP-9 siRNA treated cells. Our results demonstrate that MMP-9 targeted RNAi is able to successfully suppress MMP-9 gene expression and inhibit cell growth and invasion of SGC7901 gastric adenocarcinoma in vitro and in vivo. MMP-9 is a potential therapeutic target for gastric adenocarcinomas. PMID- 19680829 TI - Emergency medicine training: is undergraduate training sufficient for the Swiss health-care system? PMID- 19680831 TI - Management of patients with nephrotic syndrome. AB - Nephrotic syndrome is characterised by proteinuria >3.5 g/24h, oedema, hypoalbuminaemia and hyperlipidaemia. Several glomerular diseases, either primary or secondary, may lead to nephrotic syndrome. Investigations for nephrotic syndrome include immunological and infectious evaluations. Renal biopsy is often mandatory, except in diabetes. Depending on aetiology specific treatment, often with immunosuppressive agents, may be implemented. In any cases nonspecific treatment should be started with ACE inhibitors or ARBs. Urinary protein loss leads to several complications: water and sodium retention, hyperlipidaemia, increased risk of thromboembolism and infection, anaemia and alteration of mineral metabolism. Each of these complications must be identified. PMID- 19680830 TI - Swiss clinical practice guidelines for skin cancer in organ transplant recipients. AB - Patients with a solid organ transplant have increased in numbers and in individual survival in Switzerland over the last decades. As a consequence of long-term immunosuppression, skin cancer in solid organ recipients (SOTRs) has been recognized as an important problem. Screening and education of potential SOTRs about prevention of sun damage and early recognition of skin cancer are important before transplantation. Once transplanted, SOTRs should be seen by a dermatologist yearly for repeat education as well as early diagnosis, prevention and treatment of skin cancer. Squamous cell carcinoma of the skin (SCC) is the most frequent cancer in the setting of long-term immunosuppression. Sun protection by behaviour, clothing and daily sun screen application is the most effective prevention. Cumulative sun damage results in field cancerisation with numerous in-situ SCC such as actinic keratosis and Bowen's disease which should be treated proactively. Invasive SCC is cured by complete surgical excision. Early removal is the best precaution against potential metastases of SCC. Reduction of immunosuppression and switch to mTOR inhibitors and potentially, mycophenolate, may reduce the incidence of further SCC. Chemoprevention with the retinoid acitretin reduces the recurrence rate of SCC. The dermatological follow up of SOTRs should be integrated into the comprehensive post-transplant care. PMID- 19680832 TI - Emergency medicine training: a prospective, comparative study of an undergraduate clinical clerkship and an army programme. AB - OBJECTIVES: To evaluate and compare the educational impact of the University of Geneva Faculty of Medicine (UGFM) emergency medicine clerkship training with that provided by the Swiss Army medical officer cadets school (ARMY). The assessment was designed to assess students' clinical knowledge and competency in major emergency situations, ACLS (Advanced Cardiac Life Support) and ATLS (Advanced Traumatic Life Support). METHODS: Prospectively, 56 UGFM students were compared with 52 ARMY officer cadets by a multiformat pre- and post-training examination. The exam consisted of a multiple-choice questionnaire (MCQ), a standardised vignette-based oral exam (SOE) and a standardised practical cardio-pulmonary resuscitation (CPR) exercise. RESULTS: Overall, on the pre- and post-training testing, total scores improved significantly for the UGFM by 10% (from 63 to 73%) and for the ARMY by 9% (from 60 to 69%). Knowledge assessed on the MCQ improved for the UGFM by 8% (64 to 72%) but not significantly for the ARMY. Performance on the SOE improved by 10% for UGFM (54 to 64%) and the ARMY (47 to 57%) as well as performance on the CPR, which improved by 15% for UGFM (72 to 87%) and 19% for the ARMY (67 to 86%). Post-training performance indicated that, respectively, UGFM scored significantly higher than the ARMY on the MCQ (72 and 68%) and the SOE (64 and 57%) but not on the CPR. Internal reliability indexes for the MCQ, SOE and CPR were respectively 0.72, 0.86 and 0.92. Correlations between the MCQ, SOE and CPR varied between 0.07 to 0.19. CONCLUSIONS: In general, the multimethod assessment seemed to provide a complementary approach to evaluation of the trainees' competency in emergency training. Except for the ARMY MCQ performance, both training programmes seemed to be effective in improving trainees' overall knowledge and clinical performance. The trainees' performances are reviewed and discussed in terms of the specific skills assessed on the SOE, the context of the trainees' expected level of performance, the teaching and evaluation approaches, and implications in establishing the equivalence of the two programmes. PMID- 19680833 TI - Economic burden of unjustified medications at hospital discharge. AB - QUESTION UNDER STUDY: Medication errors are a major concern for health care since they may cause or lead to inappropriate medication use or patient harm. However, little is known regarding the economic burden of unjustified medications. METHODS: Hospital discharge records of 577 patients were prospectively screened for the presence of unjustified medications. From this sample population, 318 (55%) were eligible and their data were used to assess the monthly costs of unjustified discharge medications, their relationship to the total and each individual's drug expenditure, and the relative cost weights of relevant unjustified drug classes. RESULTS: The results found that 619 out of 3691 prescriptions (16.8%) were unjustified. The mean (median; 95% CI) monthly costs of unjustified discharge medications were 32 euro (27 euro; 29 euro to 35 euro). The percentage of unnecessary treatments was inversely linked to the amount of total individual drug expenditure. For this collective, monthly extra costs due to unjustified medications were 18585 euro, and the relative cost weights of the relevant drug classes were 45.8% for gastrointestinal agents (33.8% for proton pump inhibitors), 17.7% for cardiovascular drugs, and 17.2% for psychiatric drugs. CONCLUSIONS: There is a considerable financial burden imposed by unjustified medications at hospital discharge. Discharge medications not motivated by appropriate diagnoses should be questioned. This study should be repeated in other institutions and in a larger population. PMID- 19680834 TI - Intravenous leiomyomatosis of the uterus: link with new fertilisation methods? PMID- 19680835 TI - Estimated intake of the sweeteners, acesulfame-K and aspartame, from soft drinks, soft drinks based on mineral waters and nectars for a group of Portuguese teenage students. AB - In a survey of levels of acesulfame-K and aspartame in soft drinks and in light nectars, the intake of these intense sweeteners was estimated for a group of teenage students. Acesulfame-K was detected in 72% of the soft drinks, with a mean concentration of 72 mg l(-1) and aspartame was found in 92% of the samples with a mean concentration of 89 mg l(-1). When data on the content of these sweeteners in soft drinks were analysed according to flavour, cola drinks had the highest mean levels for both sweeteners with 98 and 103 mg l(-1) for acesulfame-K and aspartame, respectively. For soft drinks based on mineral water, aspartame was found in 62% of the samples, with a mean concentration of 82 mg l(-1) and acesulfame-K was found in 77%, with a mean level of 48 mg l(-1). All samples of nectars contained acesulfame-K, with a mean concentration of 128 mg l(-1) and aspartame was detected in 80% of the samples with a mean concentration of 73 mg l(-1). A frequency questionnaire, designed to identify adolescents having high consumption of these drinks, was completed by a randomly selected sample of teenagers (n = 65) living in the city of Coimbra, in 2007. The estimated daily intakes (EDI) of acesulfame-K and aspartame for the average consumer were below the acceptable daily intakes (ADIs). For acesulfame-K, the EDI was 0.7 mg kg(-1) bw day(-1) for soft drinks, 0.2 mg kg(-1) bw day(-1) for soft drinks based on mineral waters, and 0.5 mg kg(-1) bw day(-1) for nectars, representing 8.0%, 2.2%, and 5.8% of the ADI, respectively. A similar situation was observed for aspartame. In this way, the EDI for soft drinks was 1.1 mg kg(-1) day(-1), representing only 2.9% of the ADI. In respect of nectars, the EDI was 0.2 mg kg( 1) bw day(-1), representing 0.5% of the ADI. Soft drinks based on mineral waters showed the lowest EDI values of 0.3 mg kg(-1) bw day(-1), accounting for 0.7% of the ADI. PMID- 19680836 TI - Coumarin and cinnamaldehyde in cinnamon marketed in Italy: a natural chemical hazard? AB - Some plants that are processed into foods often contain natural substances that may be hazardous to human health. One example is coumarin, which is known to cause liver and kidney damage in rats, mice and probably humans. The main source of coumarin in the diet is cinnamon. The name 'cinnamon' is correctly used to refer to Ceylon cinnamon, also known as 'true cinnamon'. However, other plant species are sometimes sold with the label of cinnamon. This is the case of Cinnamomun aromaticum (cassia). In recent years, due to its cheaper price, cassia is replacing true cinnamon in the European food market being largely used in the preparation of some kinds of sweets. Several European health agencies have recently warned against consuming high amounts of cassia due to its high content of coumarin. In this study, 34 samples of cinnamon and 50 samples of cinnamon containing foodstuffs were collected from the Italian market. Quantitative determinations of coumarin and cinnamaldehyde were performed by high-performance liquid chromatography (HPLC) with diode array detector (DAD). The analytical method was in-house validated assessing recovery, repeatability, linearity, limit of detection (LOD) and limit of quantitation (LOQ). The results showed that about 51% of cinnamon samples consisted of cassia, 10% were probably a blend of cassia and Ceylon cinnamon, whereas only 39% were actually Ceylon cinnamon. As far as cinnamon-containing foods are concerned, the samples often exceeded the maximum level fixed in the European Flavourings Directive of 2 mg kg(-1). PMID- 19680837 TI - Assessment of hydroxymethylfurfural intake in the Spanish diet. AB - Estimation of hydroxymethylfurfural (HMF) exposure of the Spanish population from heat-processed food was performed. HMF levels in Spanish foods were obtained from the data previously published in peer-reviewed papers; in addition, it was identified which food categories contributed significantly to HMF exposure. The potential HMF exposure was calculated for three different scenarios by using individual food intakes and the minimum (scenario 1), median (scenario 2) and maximum (scenario 3) values of analytical data on the HMF content in foods. A mean HMF intake of 10 mg day(-1) (corresponding to scenario 2) was obtained, which is only ten-fold lower than the tolerable daily intake. Coffee and bread are the most important food items that contribute nearly 85% to the total HMF daily exposure, although biscuits, breakfast cereals, beer, UHT milk and tomato products are also important for HMF exposure. PMID- 19680838 TI - Erythromycin residue in honey from the Southern Marmara region of Turkey. AB - Honey samples, collected from the Southern Marmara region of Turkey, were analysed for erythromycin residues by liquid chromatography-mass spectrometry using electrospray ionization in the positive ion mode (LC-ESI-MS). Fifty samples, comprising chestnut, pine, linden and multi-flower honeys, were collected directly from hives and analyzed. The limit of detection and quantification were 6 and 20 ng g(-1), respectively, and recovery ranged from 85 to 89%. Four of the honey samples (8%) were found to be contaminated with erythromycin residues at concentrations ranging from 50 to 1776 ng g(-1). An erythromycin-fortified cake feeding assay was also performed in a defined hive to test the transfer of erythromycin residue to the honey matrix. In this test hive, the residue level in the honey, 3 months after dosing, was approximately 28 ng g( 1). PMID- 19680840 TI - Balancing the risks and the benefits of local fish consumption in Bermuda. AB - Fish consumption today is widely recognized as highly beneficial since it constitutes a good source of several essential nutrients, such as selenium and polyunsaturated fatty acids (n-3 PUFA). However, fish can also contain contaminants such as mercury, which make the consumer, especially pregnant women, confused about the risk-benefit balance associated with fish consumption. This is particularly true for tropical fish species for which little information is available. We have previously reported that some Bermudian neonates had elevated mercury in their umbilical blood compared with international guidelines. The objective of this study was to give precise and balanced information on the content of mercury, selenium and PUFA in the most consumed fish species in Bermuda. In 2003 and 2006, a total of 307 fish were collected from 43 fish species and 351 samples were analysed (305 flesh samples, 44 liver samples, one roe and one fat sample) by inductively coupled plasma-mass spectrometry (ICP-MS) (metals) and high-resolution gas chromatography (HRGC) (fatty acids). Results show that mercury varies among species from 0.03 to 3.3 microg g(-1) and that it is possible for at-risk groups such as pregnant women to make informed choices concerning fish consumption, e.g. maximizing fish species rich in nutrients and low in mercury. PMID- 19680839 TI - Feasibility of using capillary zone electrophoresis with photometric detection for the trace level detection of bromate in drinking water. AB - The feasibility of applying a capillary zone electrophoretic (CZE) method for the trace analysis of bromate, a suspect human carcinogen, in drinking water was studied. Using a bare fused-silica capillary (75 microm inner diameter) coupled with indirect ultraviolet detection (200 nm), 0.25 mM cetyltrimethylammonium bromide, 5% methanol and 5.0 mM phthalate at pH 5.0-5.2, an electrokinetic injection of 15 kV and 10 s, a separation voltage of 18 kV (negative polarity) and a capillary temperature of 15 degrees C, bromate was detected in high purity water at < or =10 microg l(-1). The method was applied to three bottled water sources and to local municipal water. If needed, additional sample-handling steps, consisting of an off-line pre-concentration step and pH adjustment to 5.5, was used to improve detection limits (from a high of 400 microg l(-1) to < or =10 microg l(-1)) and baseline noise. Signal-to-noise ratios also increased by adding sodium phosphate (1.1 microg ml(-1)) to all sources prior to analysis. Although the CZE method was capable of detecting bromate at levels < or =10 microg l(-1) with an analysis time of 8-9 min, high variability (>10% relative standard deviation) precluded its application to some of sources without further method development. Nonetheless, this method could serve as the basis for the detection of bromate to specific water sources with minimal or no optimization. PMID- 19680841 TI - Anatomical distribution of heavy metals in the scallop Pecten maximus. AB - This paper studied the anatomical distribution of mercury (Hg), cadmium (Cd), lead (Pb), chromium (Cr), nickel (Ni), arsenic (As), silver (Ag), copper (Cu) and zinc (Zn) in the scallop Pecten maximus and the possible implications in terms of shellfish management. Six organs were analysed: mantle, gills, foot, digestive gland, kidney and gonad. On the basis of their anatomical distribution, two groups of metals were able to be distinguished: the first included Pb, Hg, Ni, Zn and Ag; and the second comprised the four other metals studied. The metals in the first group preferentially accumulated in the kidney (except for Pb), with generally much lower concentrations in the other organs. The metals in the second group accumulated mainly in the digestive gland. As and Cu were included in the second group, but they also had particular inter-organ distribution characteristics. Among the edible organs of the scallop only the adductor muscle contained important proportions of one metal, As (which is very likely accumulated as a non-toxic derivative). A selective evisceration of the metal rich non-edible organs may therefore be considered a reliable measure to be taken with a view to reduce the metal content of scallops used for human consumption. This could be especially relevant for Cd, which is accumulated in high concentrations in the digestive gland. PMID- 19680842 TI - Effect of cooking method and rice type on arsenic concentration in cooked rice and the estimation of arsenic dietary intake in a rural village in West Bengal, India. AB - Arsenic (As) contamination of rice plants can result in high total As concentrations (t-As) in cooked rice, especially if As-contaminated water is used for cooking. This study examines two variables: (1) the cooking method (water volume and inclusion of a washing step); and (2) the rice type (atab and boiled). Cooking water and raw atab and boiled rice contained 40 microg As l(-1) and 185 and 315 microg As kg(-1), respectively. In general, all cooking methods increased t-As from the levels in raw rice; however, raw boiled rice decreased its t-As by 12.7% when cooked by the traditional method, but increased by 15.9% or 23.5% when cooked by the intermediate or contemporary methods, respectively. Based on the best possible scenario (the traditional cooking method leading to the lowest level of contamination, and the atab rice type with the lowest As content), t-As daily intake was estimated to be 328 microg, which was twice the tolerable daily intake of 150 microg. PMID- 19680844 TI - Feeding of pregnant sows with mycotoxin-contaminated diets and their non-effect on foetal and maternal hepatic transcription of genes of the insulin-like growth factor system. AB - The insulin-like growth factor (IGF) system regulates foetal and placental growth. It can be influenced by dietary factors, but little is known about the effects of mycotoxin feeding on hepatic levels of the IGF system. The aim was (1) to determine the normal foetal and maternal hepatic transcription of major IGF genes at distinct stages of pregnancy and (2) to find out if the mycotoxins deoxynivalenol (DON) and zearalenone (ZON) affect transcript concentrations. Pregnant sows were fed with naturally contaminated diets from days 35 to 70 (experiment 1), or from days 75 to 110 (experiment 2), or with control diets. Foetal hepatic IGF transcripts were determined at days 70 and 110, maternal transcripts at day 70 by quantitative polymerase chain reaction (qPCR). The highest levels of IGF-I transcripts were found in sows. On the contrary, IGF-II was predominantly expressed in foetuses at day 70. Expression of IGF-IR and IGFBP 3 decreased, whereas that of IGFBP-2 increased towards term. IGF-IIR and IGFBP-1 expression was similar in both foetuses and full-term piglets. IGF-IIR showed reduced levels in the maternal liver. The exposure of pregnant sows to DON- and ZON-contaminated diets significantly reduced the maternal and tended to reduce the weight gain of piglets in experiment 2, but had no effects on hepatic levels of IGF transcripts in both experiments. This suggests that mycotoxin-contaminated diets can induce growth depression in pigs during pregnancy without affecting hepatic transcription of major IGF genes. PMID- 19680843 TI - Co-occurrence of fumonisins with aflatoxins in home-stored maize for human consumption in rural villages of Tanzania. AB - This study determined maize-user practices that influence the presence of fumonisin and aflatoxin contamination of maize in food consumed in the rural areas of Tanzania. Samples of the 2005 maize harvest in Tanzania were collected from 120 households and examined for fumonisins and aflatoxins. Information on whether the maize was sorted to remove defective (visibly damaged or mouldy) maize before storage and whether the damaged and mouldy maize or the non-dehulled maize was used as food was also collected. In addition, the percentage of defective kernels in the samples was determined. Ninety per cent of the households sorted out defective maize, 45% consumed the defective maize and 30% consumed non-dehulled maize. In 52% of the samples fumonisins were determined at levels up to 11,048 microg kg(-1) (median = 363 microg kg(-1)) and in 15% exceeded 1000 microg kg(-1); the maximum tolerable limit (MTL) for fumonisins in maize for human consumption in other countries. Aflatoxins were detected in 18% of the samples at levels up to 158 microg kg(-1) (median = 24 microg kg(-1)). Twelve per cent of the samples exceeded the Tanzanian limit for total aflatoxins (10 microg kg(-1)). Aflatoxins co-occurred with fumonisins in 10% of the samples. The percentage defective kernels (mean = 22%) correlated positively (r = 0.39) with the fumonisin levels. Tanzanians are at a risk of exposure to fumonisins and aflatoxins in maize. There is a need for further research on fumonisin and aflatoxin exposure in Tanzania to develop appropriate control strategies. PMID- 19680845 TI - Fate of Fusarium mycotoxins in maize flour and grits during extrusion cooking. AB - Extrusion technology is used widely in the manufacture of a range of breakfast cereals and snacks for human consumption and animal feeds. To minimise consumer exposure to mycotoxins, the levels of deoxynivalenol (DON) and zearalenone (ZON) in cereals/cereal products and fumonisins B(1) and B(2) (FB(1) and FB(2)) in maize are controlled by European Union legislation. Relatively few studies, however, have examined the loss of Fusarium mycotoxins during processing. The behaviour of FB(1), FB(2) and fumonisin B(3) (FB(3)), DON and ZON during extrusion of naturally contaminated maize flour and maize grits is examined using pilot-scale equipment. DON and ZON are relatively stable during extrusion cooking but the fumonisins are lost to varying degrees. There is some loss of ZON when present in low concentrations and extruded at higher moisture contents. The presence of additives, such as reducing sugars and sodium chloride, can also affect mycotoxin levels. Moisture content of the cereal feed during extrusion is important and has a greater effect than temperature, particularly on the loss of fumonisins at the lower moistures. The effects are complex and not easy to explain, although more energy input to the extruder is required for drier materials. However, on the basis of these studies, the relationship between the concentration of Fusarium toxins in the raw and finished product is toxin- and process-dependent. PMID- 19680846 TI - Sphingoid base levels in humans consuming fumonisin-contaminated maize in rural areas of the former Transkei, South Africa: a cross-sectional study. AB - High incidences of oesophageal cancer are associated with the consumption of subsistence-grown maize by rural populations in the former Transkei region of Eastern Cape Province, South Africa. This cross-sectional study was conducted in the north-eastern magisterial area of Bizana (a previously low oesophageal cancer incidence area) and the south-eastern area of Centane (a previously high incidence area). Plasma and urine samples of male and female participants were analysed for the sphingoid bases, sphinganine and sphingosine. Good home-grown and visibly mouldy maize samples, collected from the households of the participants, were analysed for fumonisin B(1), B(2) and B(3). Plasma sphinganine/sphingosine ratios in males and females were significantly lower (p < 0.05) due to lower sphinganine levels in Bizana compared to Centane. In contrast, the urinary female and combined (males + females) sphinganine/sphingosine ratios were significantly higher (p < 0.05) in Bizana due to the significantly lower (p < 0.05) urinary sphingosine levels. Interestingly, urinary sphingoid base levels were significantly lower (p < 0.05) in males than females within each area. Based on the mean total fumonisin levels in good maize, the estimated mean probable daily intake (PDI) was 5.8 microg kg(-1) body weight day(-1) in Bizana during 2000 and 4.4 and 6.7 5.8 microg kg(-1) body weight day(-1) in Centane during 1997 and 2000, respectively, exceeding the maximum tolerable daily intake proposed by JECFA. However, there was no significant difference in the mean total fumonisin levels in the maize between the magisterial areas. The observed differences in plasma and urinary sphingoid base levels could not be ascribed as a biomarker of fumonisin exposure and further studies at an individual level are required. PMID- 19680847 TI - Mycotoxicological quality evaluation of corn samples used by processing industries in the Northern region of Parana State, Brazil. AB - Based on fungal and fumonisin contamination of 870 freshly harvested samples, the quality of corn used by processing industries in the Northern region of Parana State, Brazil (2003 and 2004 crop-year) was evaluated. Sampling was carried out for each crop at two points in the production chain, i.e. at reception by the processors and at the pre-drying step. Corn samples were more frequently contaminated with Fusarium sp. (100%) and Penicillium sp. (84.1-95.3%) than Aspergillus sp. (5.6-19.8%). Fumonisin B(1) (FB(1)) was detected in all samples from the two points in both crop-years. FB(1) levels ranged 0.02-11.83 microg g( 1) in the reception and 0.02-10.98 microg g(-1) in the pre-drying samples of the 2003 crop. Samples from the 2004 crop showed FB(1) levels ranging 0.03-12.04 microg g(-1) in the reception and 0.06-7.74 microg g(-1) in the pre-drying samples. FB(2) levels ranged 0.02-5.25 microg g(-1) in the reception and 0.01 7.89 microg g(-1) in the pre-drying samples (2003 crop-year). In samples from the 2004 crop, FB(2) levels ranged 0.02-6.12 microg g(-1) in the reception and 0.05 3.47 microg g(-1) in the pre-drying samples. Low fumonisin levels were detected in most corn samples used by processors in the Northern region of Parana State, showing a decreasing trend in fumonisin contamination over the years. PMID- 19680848 TI - Increased aflatoxin contamination of dried figs in a drought year. AB - Dried figs (4917 samples) destined for export from Turkey to the European Union were collected between September and December during the very dry crop year of 2007 and tested for aflatoxins B(1), B(2), G(1) and G(2) by immunoaffinity column clean-up and reverse-phase high-performance liquid chromatography (RP-HPLC). While 32% of the samples contained detectable levels of total aflatoxins, 9.8% of them exceeded the European Union limits. Aflatoxin levels were in the range of 0.2-259.46 microg kg(-1) and 2.04-259.46 microg kg(-1) for all samples and samples that exceeded the limits, respectively. A substantial increase in the incidence of aflatoxins was observed in 2007 compared with previous years, most likely due to the drought stress, high temperatures and low relative humidity encountered during the period from January to September of that year. In 2007, the mean temperature was 1-2 degrees C higher, there was 300 mm less total rain, and the mean relative humidity was 10-15% lower than in 2002-06. The average concentration of individual aflatoxins present in the samples was quantified to determine whether the drought conditions promoted certain types of aflatoxins. Among the contaminated samples, aflatoxin B(1) occurred in 97% of the contaminated samples, followed by G(1) in 47%, B(2) in 24%, and G(2) in 6% of samples. Concentrations of individual aflatoxins exhibited great variability among the samples but were not significantly different from those reported in previous studies, which were conducted under conditions without drought and high temperatures. PMID- 19680849 TI - Diffusion behaviour of additives in polypropylene in correlation with polymer properties. AB - The migration behaviour of polymer additives in 17 polypropylene (PP) samples is described. These samples cover the major types of PP used in food packaging. The diffusion coefficients of additives with relatively small molecular masses, M(r) = 136 (limonene), as well as the migration of typical antioxidants used in PP up to M(r) = 1178 (IRGANOX 1010), were measured at different temperatures. In addition, the diffusion data and percentages of xylene-soluble fractions were correlated. This enables a prediction of the migration behaviour of a PP sample by testing its 'isotactic index' with xylene. The results clearly indicate that PP can be subdivided, from the migration point of view, into the monophasic homopolymer (h-PP), the monophasic random copolymer (r-PP), and the heterophasic copolymer (heco-PP). The diffusion coefficients for r-PP are at least one order of magnitude higher than those of h-PP and comparable with the values for heco PP. Upper limits for the diffusion values can be calculated based on the safety margin required by consumer protection laws. PMID- 19680850 TI - Determination of 2,6-diisopropylnaphthalene (DIPN) and n-dibutylphthalate (DBP) in food and paper packaging materials from US marketplaces. AB - A gas chromatography-ion-trap tandem mass spectrometry procedure was developed for the determination of 2,6-diisopropylnaphthalene (DIPN) and n-dibutylphthalate (DBP) in domestic and imported paper packages and food sold in US marketplaces. The procedure involved ultrasonic extraction with dichloromethane, followed by analysis with the gas chromatography-ion-trap tandem mass spectrometry. Calibration curves for DIPN and DBP were achieved with concentrations ranging from 0.01 to 10 microg ml(-1) and the corresponding r(2) values were 0.9976 and 0.9956, respectively. In most of the fortified samples the recoveries were higher than 80% with a relative standard deviation (RSD) <10%. Using this procedure, it was found that less than 20% of the tested domestic packages and more than 60% of the tested imported food packages contained both DIPN and DBP. The concentrations of DIPN and DBP ranged from 0.09 to 20 mg kg(-1) and 0.14 to 55 mg kg(-1), respectively, with most of the DINP and DBP levels lower than 20 mg kg(-1). DIPN was not detected (<0.01 mg kg(-1)) in 41 food samples and DBP was only detected in two domestic and four imported food samples with concentrations ranging from <0.01 to 0.81 mg kg(-1). PMID- 19680851 TI - Development and characterization of an active polyethylene film containing Lactobacillus curvatus CRL705 bacteriocins. AB - The development and characterization of a bacteriocin-containing polyethylene based film is described, incorporating lactocin 705 and lactocin AL705, produced by Lactobacillus curvatus CRL705, and nisin. Three different procedures to obtain lactocin 705 and AL705 solution were evaluated, with the partially purified aqueous bacteriocin solution showing the highest inhibitory activity against indicator strains (Lactobacillus plantarum CRL691 and Listeria innocua 7). Pouch contact, soaking and a contact method were compared for incorporating bacteriocins onto PE-based films. Contact between the PE film and bacteriocin solution was the most effective, resulting in a more uniform distribution of bacteriocins on the film surface and using less active solution. The minimal inhibitory concentration of bacteriocin solution was 267 AU cm(-3) (lactocin 705) and 2133 AU cm(-3) (lactocin AL705), while the minimal contact time was 1 h. When relative inhibition area for antilisterial activity of the active films was compared, those treated with L. curvatus CRL705 bacteriocins displayed higher inhibitory activity than nisin-treated films. Functional properties of active PE films containing lactocin 705 and AL705 showed no differences compared with non active control films. Bacteriocin-active PE-based films are shown to be highly effective in inhibiting growth of Listeria. The potential use of commercially available packaging films as bacteriocins carriers may benefit active-packaging systems. PMID- 19680853 TI - Development of veterinary drug residue controls by the Codex Alimentarius Commission: a review. AB - Within the Codex Alimentarius, transparency is and has been an important consideration with regard to how and why actions and decisions are made in providing stakeholders with sufficient information and clarity on matters of interest and decision-making. This review focuses on the development of analytical methods performance criteria for measuring veterinary drug residues in food-producing animals for the Codex Committee on Residues of Veterinary Drugs in Foods. It describes the initial development as well as the evolution and refinement of analytical method performance criteria as well as method validation criteria from its first meeting to the present. Reference is routinely made to published Codex, Food and Agriculture of the United Nations (FAO) reports, guidance documents and manuals as well as those of other relevant analytical method standard-setting organizations with regard to analytical method performance relevant to Codex Alimentarius food safety standards development. Key Codex decisions that have substantially impacted on analytical methods-related issues are described and the subsequent outcomes. It is a reasoned conclusion that the refinement of analytical method performance criteria and validation is an ongoing process that must continuously adapt to present and future scientific issues and decisions within the Codex Alimentarius Commission and its subsidiary bodies as well as relevant analytical standard-setting organizations. PMID- 19680854 TI - Comparison of methods for the estimation of measurement uncertainty for an analytical method for sulphonamides. AB - A simple and inexpensive liquid chromatographic method for the determination of seven sulphonamides in animal tissues was validated. The measurement uncertainty of the method was estimated using two approaches: a 'top-down' approach based on in-house validation data, which used either repeatability data or intra laboratory reproducibility; and a 'bottom-up' approach, which included repeatability data from spiking experiments. The decision limits (CCalpha) applied in the European Union were calculated for comparison. The bottom-up approach was used to identify critical steps in the analytical procedure, which comprised extraction, concentration, hexane-wash and HPLC-UV analysis. Six replicates of porcine kidney were fortified at the maximum residue limit (100 microg kg(-1)) at three different stages of the analytical procedure, extraction, evaporation, and final wash/HPLC analysis, to provide repeatability data for each step. The uncertainties of the gravimetric and volumetric measurements were estimated and integrated in the calculation of the total combined uncertainties by the bottom-up approach. Estimates for systematic error components were included in both approaches. Combined uncertainty estimates for the seven compounds using the 'top-down' approach ranged from 7.9 to 12.5% (using reproducibility) and from 5.4 to 9.5% (using repeatability data) and from 5.1 to 9.0% using the bottom-up approach. CCalpha values ranged from 105.6 to 108.5 microg kg(-1). The major contributor to the combined uncertainty for each analyte was identified as the extraction step. Since there was no statistical difference between the uncertainty values obtained by either approach, the analyst would be justified in applying the 'top-down' estimation using method validation data, rather than performing additional experiments to obtain uncertainty data. PMID- 19680855 TI - AFNOR validation of Premi Test, a microbiological-based screening tube-test for the detection of antimicrobial residues in animal muscle tissue. AB - Premi Test contains viable spores of a strain of Bacillus stearothermophilus which is sensitive to antimicrobial residues, such as beta-lactams, tetracyclines, macrolides and sulphonamides. The growth of the strain is inhibited by the presence of antimicrobial residues in muscle tissue samples. Premi Test was validated according to AFNOR rules (French Association for Normalisation). The AFNOR validation was based on the comparison of reference methods (French Official method, i.e. four plate test (FPT) and the STAR protocol (five plate test)) with the alternative method (Premi Test). A preliminary study was conducted in an expert laboratory (Community Reference Laboratory, CRL) on both spiked and incurred samples (field samples). Several method performance criteria (sensitivity, specificity, relative accuracy) were estimated and are discussed, in addition to detection capabilities. Adequate agreement was found between the alternative method and the reference methods. However, Premi Test was more sensitive to beta-lactams and sulphonamides than the FPT. Subsequently, a collaborative study with 11 laboratories was organised by the CRL. Blank and spiked meat juice samples were sent to participants. The expert laboratory (CRL) statistically analysed the results. It was concluded that Premi Test could be used for the routine determination of antimicrobial residues in muscle of different animal origin with acceptable analytical performance. The detection capabilities of Premi Test for beta-lactams (amoxicillin, ceftiofur), one macrolide (tylosin) and tetracycline were at the level of the respective maximum residue limits (MRL) in muscle samples or even lower. PMID- 19680857 TI - Validation of radioimmunoassay screening methods for beta-agonists in bovine liver according to Commission Decision 2002/657/EC. AB - Validation studies were carried out on a multi-residue screening method for anilinic type beta-agonists (clenbuterol, mabuterol, brombuterol, cimaterol, cimbuterol, mapenterol, clenpenterol) and a method for the phenolic type beta agonist, salbutamol, in bovine liver. The validation was performed according to the European Union Commission Decision 2002/657/EC (European Commission 2002), which establishes criteria and procedures for the determination of parameters such as the detection capability (CCbeta), specificity, stability of standard solutions and stability of the analyte in matrix. CCbeta values for the eight target compounds were between 0.25 and 0.5 microg kg(-1). The stability of standard solutions and analytes in matrix and the specificity of the antibody were characterized. The methods are applicable for qualitative screening of beta agonists for regulatory programmes according to European Union performance requirements, or as a semi-quantitative research tool for known target compounds. PMID- 19680858 TI - Determination of the performance characteristics of a new multi-residue method for non-steroidal anti-inflammatory drugs, corticosteroids and anabolic steroids in food animal tissues. AB - A new LC-MS/MS method was developed for the analysis of 29 veterinary drug residues, spanning three different drug classes, in animal tissues. The procedures used to measure the characteristic performance parameters of the method and the results obtained using fortified blank bovine muscle and kidney tissue are described. For a quantitative and confirmatory method, the characteristic performance parameters to be determined are the limits of quantification, trueness, recovery, precision, selectivity, ruggedness, and stability. The characteristic performance parameters defined for the method will be verified during a validation study by an independent experienced analyst to determine whether the method is suitable for use in a regulatory monitoring and control program for residues of the 29 analytes. PMID- 19680856 TI - Collaborative study of a microbiological screening method (three-plate) for the banned antimicrobial growth promotors tylosin, virginiamycin, spiramycin, zinc bacitracin and avoparcin in animal feed. AB - A microbiological screening method (three-plate) for the detection of the antimicrobial growth promoters tylosin, spiramycin, virginiamycin, zinc bacitracin, and avoparcin in animal feed has been developed and validated successfully. A collaborative study involving 18 laboratories receiving 172 samples was carried out to verify the performance characteristics. The detection level for tylosin/virginiamycin/spiramycin, expressed in microbiological activity, was 1 mg kg(-1) (false-positives, 2%; false-negatives, 3, 0, and 6%, respectively). Avoparcin could be detected at 1 mg kg(-1) in feed in general (false-positives, 2%; false-negatives, 0%). However, in calf feed the sensitivity was lower. The percentages of false-negatives were found to be 12%, 7%, and 0% at 1, 3, and 5 mg kg(-1), respectively (false-positives, 4%). The limit of detection for zinc bacitracin was 3-5 mg kg(-1) (false-positives, 5-10%; false-negatives, 77% at 1 mg kg(-1), 45% at 2 mg kg(-1), 12% at 3 mg kg(-1), and 4% at 5 mg kg( 1)). The method allowed for a distinction to be made between the groups of antibiotics: avoparcin/zinc bacitracin versus tylosin/virginiamycin/spiramycin. This definitely gives added value to the method in the framework of a follow-up of positive screening results by post-screening and confirmatory analysis. PMID- 19680859 TI - Determination of ibuprofen, ketoprofen, diclofenac and phenylbutazone in bovine milk by gas chromatography-tandem mass spectrometry. AB - A method has been developed to analyse for ibuprofen (IBP), ketoprofen (KPF), diclofenac (DCF) and phenylbutazone (PBZ) residues in bovine milk. Milk samples were extracted with acetonitrile and sample extracts were purified on Isolute C(18) solid-phase extraction cartridges. Aliquots were analysed by gas chromatography-tandem mass spectrometry (GC-MS/MS). The method was validated in bovine milk, according to the criteria defined in Commission Decision 2002/657/EC. The decision limits (CCalpha were 0.59, 2.69, 0.90 and 0.70 ng ml( 1), respectively, for IBP, KPF, DCF and PBZ, and detection capabilities (CCbeta) of 1.01, 4.58, 1.54 and 1.19 ng ml(-1), respectively, were obtained. The measurement uncertainty of the method was 17.8%, 80.9%, 28.2% and 20.2% for IBP, KPF, DCF and PBZ, respectively. Fortifying bovine milk samples (n = 18) in three separate assays show the accuracy of the method to be between 104% and 112%. The precision of the method, expressed as relative standard deviations for the within laboratory reproducibility at the three levels of fortification (5, 7.5 and 10 ng ml(-1)) was less than 8% for IBP, DCF and PBZ, respectively. Poor precision was obtained for KPF with a relative standard deviation of 28%. PMID- 19680860 TI - Multi-residue quantitation of aminoglycoside antibiotics in kidney and meat by liquid chromatography with tandem mass spectrometry. AB - Quantitative methods using liquid chromatography coupled with tandem mass spectrometry were developed for seven kinds of aminoglycoside antibiotics in kidney and muscle tissues. Mass spectral acquisition was performed in the positive-ion mode by applying multiple reaction monitoring. Liquid chromatographic separation employed a ZIC-HILIC column (SeQuant) for hydrophilic interaction chromatography. Extraction of the aminoglycosides was performed using liquid extraction with a phosphate buffer containing trichloroacetic acid, followed by a solid-phase clean-up procedure on a weak cation-exchange column with carboxypropyl (CBX) SPE cartridge (Mallinckrodt Baker). The limits of quantification were 25 ng g(-1) for gentamicin, 50 ng g(-1) for spectinomycin, dihydrostreptomycin, kanamycin and apramycin, and 100 ng g(-1) for streptomycin and neomycin. These are well below the maximum residue limits set by the Codex Alimentarius Commission. The recoveries of all compounds from all tissues fortified at the level of quantification limits of 500 and 1000 ng g(-1) were >70%, and the variability (relative standard deviation) was generally <12%. PMID- 19680861 TI - Identification of hormone esters in injection site in muscle tissues by LC/MS/MS. AB - The detection of hormone abuse for growth promotion in food animal production is a global concern. Initial testing for hormones in Canada was directed at the compounds approved for use in beef cattle, melengestrol acetate, trenbolone acetate and zeranol, and the banned compound diethylstilbestrol (DES). No hormonal growth promoters are approved for use in veal production in Canada. However, instances of use of trenbolone and clenbuterol were detected in Canada in the 1990s. During the development of a new analytical method for testosterone and progesterone, there were reports of suspicious injection sites being found in veal calves. Upon implementation of the method, analysis of investigative samples revealed significant residues of testosterone in some injection sites. To prove that the source of these residues was exogenous, a fully validated method for hormone esters was developed to confirm the presence of exogenous hormones in these injection sites. The QUECHERS model was employed in methods development and resulted in a simple, effective extraction technique that consisted of sample pre homogenization, liquid/liquid partitioning, extract dilution, filtration and use of LC/MS/MS to provide detection selectivity. The result was an adaptable MS/MS confirmation technique that meets the needs of Canadian regulatory authorities to confirm the misuse of injectable testosterone, and potentially other hormones, in food animal production. PMID- 19680862 TI - Characterisation of biotoxins produced by a cyanobacteria bloom in Lake Averno using two LC-MS-based techniques. AB - Cyanobacteria (blue-green algae) cause blooms in eutrophic lakes and drinking water reservoirs. They also produce biotoxins, including microcystins (MCs), highly toxic cyclic heptapeptides that cause poisoning in animals and human. In this paper, we present a method for the analysis of four MCs by ion trap LC-MS and MALDI-TOF/MS. The data are compared to evaluate the performance and reliability of the different MS detection systems. The method was applied to the analysis of water and algae samples from Lake Averno, near Naples, as a consequence of a cyanobacteria bloom. The analysis of algae cell extracts showed no contamination by known microcystins, but the three main substances were detected. MALDI-TOF/MS was successful for screening of the biotoxins in the samples, identifying anabaenopeptin B and anabaenopeptin F as the major contaminants on the basis of literature mass spectrometry data. The structure of the third compound was not identified and is under further investigation. The method could characterise the biotoxins produced in Lake Averno for evaluating health risks related to their presence. PMID- 19680864 TI - Modelling consumer exposures to nutrients and additives in animal products. AB - Current guidelines for assessing human exposures to nutrients and other substances used in animal feed are based on methods used for veterinary pharmaceuticals. These methods assume high-level daily consumption of animal products and do not take account of differences between species or between consuming humans. A more detailed dietary modelling approach is described, which takes these factors into account as well as high-level consumption by different age groups. The proposed approach is evaluated in three case studies, iodine, selenium and astaxanthin, which have previously undergone thorough evaluation by EU authorities. When applied in a tiered approach, the proposed model provides results that are consistent with previous assessments and with results obtained using other modelling techniques. The results indicate that the method has the potential to provide a conservative, reliable and flexible alternative to existing approaches to intake estimation. PMID- 19680865 TI - Dietary exposure of secondary school students in Hong Kong to benzoic acid in prepackaged non-alcoholic beverages. AB - This study evaluated the dietary exposure of secondary school students in Hong Kong to benzoic acid from pre-packaged non-alcoholic beverages. Exposure was estimated using local food consumption data of secondary school students obtained by a semi-quantitative food frequency questionnaire in 2000 and the benzoic acid level detected in pre-packaged beverages, including soft drink (both diet/light and regular types), fruit juice, soy milk, Chinese tea and coffee/tea) available locally in late 2006. The estimated dietary exposure to benzoic acid from pre packaged beverages of average and high consumers (95(th) percentile) was 0.31 and 0.97 mg kg(-1) bw day(-1), respectively. These exposures accounted for 6.1 and 19.3% of the acceptable daily intake (ADI: 0-5 mg kg(-1) bw) of benzoic acid for average and high consumers, respectively. As in other countries, soft drinks contributed most to dietary exposure to benzoic acid from pre-packaged beverages in Hong Kong. PMID- 19680867 TI - Stability of added iodine in processed cereal foods. AB - The stability of iodine from iodized salt was measured in white bread, grain bread, sweet biscuits and the breakfast cereals, Weetbix (a flaked, pressed, wheat product), Ricies (a puffed rice product) and toasted muesli, as part of the New Zealand Government's initiatives to address the public health issue of low iodine intake by most New Zealanders. Triplicate runs of each product were manufactured by commercial food manufacturers between September 2006 and May 2007 with iodized and non-iodized salt. Triplicate samples were taken at various steps during processing of each food and analysed for moisture and iodine content. Iodine concentration was determined by inductively coupled plasma-mass spectrometry following an alkaline digestion. Iodine, at the equivalent of 25-65 mg of iodine per kg salt, was 100% retained in each of the selected foods from the time of mixing to the final product. These results imply that all iodine added via salt at the time of manufacture is available for consumption but not necessarily bioavailable. These data can be used for modelling the impact of strategies to increase iodine exposure. Salt as an ingredient is not a good predictor of iodine intake due to the inhomogeneity of iodine in iodized salt. PMID- 19680866 TI - Determination of bixin and norbixin in meat using liquid chromatography and photodiode array detection. AB - The development of an analytical method that enables routine analysis of annatto dye, specifically bixin and norbixin, in meat tissue is described. Liquid-solid extraction was carried out using acetonitrile. Analysis was by HPLC with photodiode array detection using two fixed wavelengths (458 and 486 nm). The possibilities of ion trap mass spectrometry (MS) were also assessed. Method performance characteristics, according to Commission Decision 2002/657/EC, were determined, with recoveries between 99 and 102% and calibration curves being linear in the 0.5-10 mg kg(-1) range. The limit of quantification was 0.5 mg kg( 1). PMID- 19680868 TI - Haloanisole and halophenol contamination in Spanish aged red wines. AB - This is the first study to be carried out on the incidence of halophenols and haloanisoles, including trichloroanisole, in aged red wines. A total of 966 red wines, aged for 6, 12 and 24 months in oak barrels and from different Spanish production areas, were analysed by stir bar sorptive extraction followed by gas chromatography/mass spectrometry (GC/MS). From the wines sampled, 155 (16.1%) were contaminated with one or several compounds, with 7.6, 6.9 and 1.5% corresponding to the 12- (aged-12), 6- (aged-6) and 24-month-aged (aged-24) wines, respectively. The most abundant compounds causing taint were 2,3,4,6 tetrachloroanisole and 2,4,6-trichloroanisol (6.8 and 5.3%, respectively). No 2,4,6-tribromophenol was found in any of the samples. Contamination with halo compounds was highest in samples from South-West Spain, followed by those from Northern Spain. The mean concentration for all compounds were always higher than their respective olfactory threshold, but none of these halo compounds represent a health hazard to humans through the consumption of commercial red aged wines. PMID- 19680869 TI - Determination of fluoroquinolones in aquaculture products by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). AB - An ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for the determination of fluoroquinolones-ciprofloxacin (CIPRO), danofloxacin (DANO), enrofloxacin (ENRO) and sarafloxacin (SARA)-in aquaculture products, specifically salmon, shrimp and tilapia. After initial sample extraction with an acidic acetonitrile solution, the extract was diluted with dichloromethane and centrifuged, then an aliquot was concentrated and applied to a C18 solid-phase extraction cartridge and concentrated for a second time. The resultant residue was dissolved in acetonitrile, diluted with water, and then further defatted with hexane. The fluoroquinolone residues were determined by UPLC with an HSS T3 C18 reverse-phase column using an ammonium hydroxide-formic acid buffer in an acetonitrile gradient with MS/MS detection using multiple reaction monitoring. Average recoveries for salmon tissue ranged from 73% for DANO to 95% for SARA, for shrimp from 71% for DANO to 109% for SARA, and from 62% for DANO to 111% for SARA in tilapia, fortified at the 1.0 ng g(-1) level. Standard curves were linear between 0.002 and 0.5 ng injected for all compounds. Detection limits of 0.2 ng g(-1) for CIPRO, DANO, ENRO, and SARA were easily obtainable. The operational errors, interferences, and recoveries for fortified samples demonstrate that this described method is suitable for routine use in a regulatory programme. The recommended method is simple, rapid, specific and reliable for the routine monitoring of fluoroquinolone residues in aquatic species such as salmon, tilapia and shrimp. PMID- 19680870 TI - Semicarbazide is non-specific as a marker metabolite to reveal nitrofurazone abuse as it can form under Hofmann conditions. AB - Detecting illegal nitrofurazone treatment of food-producing animals has been undermined by the reliance on semicarbazide as a marker metabolite for analysis because, during processing, semicarbazide forms in a variety of foods not exposed to nitrofurazone by previously unexplained pathways. In this study, formation of semicarbazide was examined under dairy product processing conditions. Hypochlorite dosed into milk on an industrial scale, at concentrations extreme for unintentional residues, produced monochloramine, but, without pH adjustment, hypochlorite alone did not generate semicarbazide. Dosed hypochlorite and peroxyacetic acid each generated low ng g(-1) levels of semicarbazide in milk (and whey) only when used in conjunction with localised high pH conditions, which can occur during both anion exchange and neutralisation. Dosed hydroxyurea generated semicarbazide at any pH, with greatest levels formed at high pH. Detecting nitrofurazone abuse can be better accomplished by analysis either for nitrofurazone itself or for the marker metabolite 5-nitro-2-furaldehyde, rather than semicarbazide. PMID- 19680872 TI - Validation of the procedure for the determination of maduramicin in concentrates, premixes, and feeds by liquid chromatography. AB - A single laboratory validation was carried out for the determination of maduramicin in concentrates, premixes, and feed. The method comprised sample extraction of maduramicin, derivatization with dansylhydrazine and liquid chromatography with ultraviolet light detection. The limit of detection (LOD) and limit of quantification (LOQ) were 0.4 and 1.0 mg kg(-1), respectively. The repeatability expressed as the average difference between the results of duplicate measurements was 5.9% at the concentration level of 1% (concentrate), 7.1% at the concentration level of 1 g kg(-1) (premix), and 11% with the feed containing maduramicin with the nominal concentration of 5 mg kg(-1) and feed spiked at the concentration level of 1 mg kg(-1). The relative standard deviations for the within-laboratory reproducibility (RSD(W)) were 9.2%, 16%, 18%, and 17% at the concentration levels of 1%, 1 g kg(-1), 5 mg kg(-1), and 1 mg kg(-1), respectively. The measurement uncertainties were +/-0.2%, +/-0.3 g kg( 1), +/-1.9 mg kg(-1), and +/-0.3 mg kg(-1) at the same concentration levels, respectively. PMID- 19680873 TI - Influence of post-harvest treatments with fludioxonil and soy lecithin co application in controlling blue and grey mould and fludioxonil residues in Coscia pears. AB - The residue levels of fludioxonil (FLU) were determined in Coscia pear following a 1-, 2- or 4-min dip in an aqueous mixture of FLU containing 300 or 100 mg l(-1) (active ingredient, a.i.) at 20 and 50 degrees C, respectively, with or without 2% soy lecithin. The efficacy of heat treatment with water and FLU mixtures was investigated on artificially inoculated pears for the control of post-harvest decay caused by blue (Penicillium expansum Link) and grey (Botrytis cinerea Pers. ex Fr.) mould. Treatment with 300 mg l(-1) FLU at 20 degrees C increased residues significantly when treatment time rose from 1 to 2 min; no further increase was recorded when dip time raised from 2 to 4 min. FLU residue rates were unaffected by treatment time when 300 mg l(-1) a.i. was applied in combination with lecithin at 20 degrees C. While treatment with 100 mg l(-1) a.i. at 50 degrees C for 1 and 2 min resulted in similar residue levels, significantly higher residues were detected when dip time increased from 1 to 4 min. Co-application of lecithin significantly decreased FLU residues with respect to fruit treated with FLU alone. Treatments with FLU at 20 or 50 degrees C effectively controlled decay over 10 days of incubation. While co-application of lecithin did not affect the efficacy of FLU at 300 mg l(-1)and 20 degrees C, treatment efficacy decreased when lecithin was applied in combination with 100 mg l(-1) FLU and 50 degrees C for 4 min and to a greater extent when dip time was 1-2 min. PMID- 19680871 TI - Thermal stability of antiparasitic macrocyclic lactones milk residues during industrial processing. AB - The chemical stability of residues of different antiparasitic macrocyclic lactone compounds in milk subjected to thermal treatment was assessed. Concentrations of ivermectin (IVM), moxidectin (MXD) and eprinomectin (EPM) in sheep milk, equivalent to those measured in vivo in milk excretion studies, were subjected to 65 degrees C over 30 min or to 75 degrees C for 15 s. Residue concentrations of IVM, MXD and EPM in milk were measured by high-performance liquid chromatography (HPLC) (fluorescence detection) before and after heat treatment of the drug fortified milk samples. No evidence of chemical loss was obtained in either of the thermal treatments under evaluation. The stability of the parent compounds in milk was evidenced by the lack of bioconversion products (metabolites) after both thermal treatments. Only very minor changes on drug concentrations were observed at the end of the treatments, which fell within the limits of the variation of the validated analytical method. In conclusion, residue concentrations of macrocyclic lactones are unaffected by industrial-simulated milk thermal procedures. Based on the reported findings, it can be postulated that residue concentrations of IVM, MXD and EPM measured in raw sheep milk may be used to estimate consumer exposure and dietary intake for these veterinary drugs. PMID- 19680874 TI - Purification of Candida guilliermondii and Pichia ohmeri killer toxin as an active agent against Penicillium expansum. AB - An antifungal assay with cell-free culture supernatant of Pichia ohmeri 158 and Candida guilliermondii P3 was tested against Penicillium expansum strain #2 at 25 degrees C by measuring hyphal length and percentage conidia germination. C. guilliermondii was more effective against P. expansum conidia germination (58.15% inhibition), while P. ohmeri showed higher inhibition of mycelial growth (66.17%), indicating a probable mechanism associated with killer activity. This killer toxin (molecular mass <3 kDa) was partially purified by normal phase HPLC, using TSKgel Amide-80 analytical and preparative columns. Compared with crude extract, the killer toxin eluted from the post analytical column significantly inhibited P. expansum:% inhibition rose from 42.16 to 90.93% (C. guilliermondii) and 39.32 to 91.12% (P. ohmeri) (p < 0.05). The one-step purification process was adequate in isolating killer toxin from culture supernatant and also increased anti-Penicillium activity. PMID- 19680875 TI - Agaritine content of 53 Agaricus species collected from nature. AB - Fifty-three different species of the genus Agaricus were collected in the Czech Republic during the period 1998-2001 and identified by an experienced mycologist. The samples were analysed for agaritine (N2-(gamma-L-glutamyl)-4 hydroxymethylphenylhydrazine) content, a precursor to a suspected rodent carcinogen. There was a huge variation in agaritine content between species, but less variation between samples of a species. Whereas the cultivated mushroom Agaricus bisporus commonly contain 200-500 mg agaritine kg(-1) fresh weight, no less than 24 of the 53 species contained agaritine levels above 1000 mg kg(-1) fresh weight. The highest level was found in A. elvensis containing up to 10, 000 mg kg(-1) fresh weight. Twenty species contained intermediate levels (100-1000 mg kg(-1)), and nine species were below 100 mg kg(-1). Some of the species producing low levels of agaritine might be candidates for future strain development of Agaricus mushrooms for cultivation. No correlation could be observed between agaritine content and size of the mushroom, week of the year when collected, year of collection, or site of collection. Besides occurring in the genus Agaricus, some species of the genera Leucoagaricus and Macrolepiota were also shown to contain agaritine. PMID- 19680876 TI - Fumonisin B2 production by Aspergillus niger in Thai coffee beans. AB - During 2006 and 2007, a total of 64 Thai dried coffee bean samples (Coffea arabica) from two growing sites in Chiangmai Province and 32 Thai dried coffee bean samples (Coffea canephora) from two growing sites in Chumporn Province, Thailand, were collected and assessed for fumonisin contamination by black Aspergilli. No Fusarium species known to produce fumonisin were detected, but black Aspergilli had high incidences on both Arabica and Robusta Thai coffee beans. Liquid chromatography (LC) with high-resolution mass spectrometric (HRMS) detection showed that 67% of Aspergillus niger isolates from coffee beans were capable of producing fumonisins B(2) (FB(2)) and B(4) when grown on Czapek Yeast Agar with 5% NaCl. Small amounts (1-9.7 ng g(-1)) of FB(2) were detected in seven of 12 selected coffee samples after ion-exchange purification and LC-MS/MS detection. Two samples also contained FB(4). This is the first record of freshly isolated A. niger strains producing fumonisins and the first report on the natural occurrence of FB(2) and FB(4) in coffee. PMID- 19680877 TI - Purification of patulin from Penicillium expansum culture: high-speed counter current chromatography (HSCCC) versus preparative high-performance liquid chromatography (prep-HPLC). AB - Patulin is a mycotoxin produced by species of Penicillium and Aspergillus and is toxic to a wide range of organisms, including humans and livestock. To produce large amount of pure patulin for research purposes, high-speed counter-current chromatography (HSCCC) and preparative high-performance liquid chromatography (prep-HPLC) were applied to the purification of patulin. Apple juice was inoculated with P. expansum and containing 0.5 mg patulin per ml was used as a starting material for separation. For HSCCC, a biphasic solvent system consisted of ethyl acetate-hexane-pH 4 acetic acid (7.5:2.5:10, v/v/v) was used. For prep HPLC, the separation was carried out in a C18 reversed-phase preparative column with a mobile phase containing acetonitrile-pH 4 acetic acid (5:95, v/v). Fractions containing patulin were collected and analysed by analytical HPLC and identified by congruent retention time and ultraviolet/visible (UV-VIS) spectrum of the standard. The structure of the purified patulin was confirmed by mass spectrometry and nuclear magnetic resonance. HSCCC produced 21.9 mg of patulin from 50 ml apple juice culture whereas the prep-HPLC yielded 18.1 mg. HSCCC also produced purer patulin than the prep-HPLC (98.6 versus 96.3%) and higher recovery (86.2 versus 71.3%). In addition, the HSCCC method is advantageous for its lower cost and a simpler procedure compared with the prep-HPLC. This one-step HSCCC method can potentially provide a simple, effective and environmentally friendly tool for obtaining gram-level pure patulin for toxicology, detoxification and many other patulin-related studies. PMID- 19680878 TI - Aflatoxin-albumin adducts and correlation with decreased serum levels of vitamins A and E in an adult Ghanaian population. AB - A study of aflatoxin (AF) exposure and the levels of vitamins A and E was carried out with a group of 507 Ghanaian participants. AFB(1)-albumin adducts (AFB-AA) were measured by radioimmunoassay and vitamins A and E were measured by high performance liquid chromatography (HPLC). The average level of serum AFB-AA was 0.94 +/- 0.64 (range = 0.1-4.44) pmol mg(-1) albumin. Mean levels of vitamins A and E were 1.32 +/- 0.48 (range = 0.41-4.85) micromol l(-1) and 15.68 +/- 4.12 (range = 6.35-30.40) micromol l(-1), respectively. A significantly negative correlation was found between serum AFB-AA and vitamin A levels (r = -0.110, p = 0.013). An even stronger, significant negative, correlation was found between serum AFB-AA and vitamin E levels (r = -0.149, p < 0.001). Serum AFB-AA levels were statistically higher (median = 0.985 pmol mg(-1) albumin) in subjects who had low levels of both vitamins A and E as compared with the levels (median = 0.741 pmol mg(-1) albumin) subjects who had high vitamins A and E levels (p(trend) = 0.001). To verify these findings, blood samples were again collected from 165 of the 507 people 3 months after the initial collection. Significantly negative correlations were confirmed between levels of serum AFB-AA and both vitamins A (r = -0.232, p = 0.003) and E (r = -0.178, p = 0.023). Again, high serum AFB-AA concentrations (median = 1.578 pmol mg(-1) albumin) were found in subjects with low levels of vitamins A and E compared with the concentrations (median = 1.381 pmol mg(-1) albumin) in subjects with high levels of vitamins A and E (p(trend) = 0.002). These data show that AF exposure was associated with decreased levels of serum vitamins A and E in high-risk human populations, which may significantly influence the incidence of AF-related adverse health effects. PMID- 19680879 TI - Effects of live yeast cell supplementation to high concentrate diets on the toxicokinetics of ochratoxin A in sheep. AB - Previous studies have indicated that high concentrate feeding reduces the ruminal degradation of the mycotoxin ochratoxin A (OA) to the less-toxic ochratoxin alpha (Oalpha) in ruminants. This is due to a pH-induced decrease in ruminal protozoa and subsequent increasing transfer of OA into the systemic circulation. The present study investigated whether stabilization of rumen pH by the live yeast cell supplementation to high concentrate diets affects the toxicokinetics of OA in sheep. Sheep were fed diets consisting of 70% concentrates and 30% grass silage (dry matter basis) supplemented without or with live yeast cells (Saccharomyces cerevisiae CNCM I-1077). After an adaptation period of 3 weeks, animals were given a single dose of OA (2.46 mg) in the form of contaminated wheat. Even though live yeast cells accelerated the recovery of ruminal pH from the decrease in pH induced by feeding, no effect on ruminal degradability and systemic availability of OA was recorded. Based on in vitro studies, live yeast cells and extracts of live yeast cell walls have been suggested as a mycotoxin binding agent. However, supplementation with live yeast cells had no effect on the excretion pattern of OA in sheep, indicating that binding of OA to yeast components may be limited in ruminants. With respect to the toxicokinetics of OA, our results are in agreement with earlier results demonstrating that the hydrolysis of OA in the gastrointestinal tract of sheep is substantially less than previously described, especially if OA is ingested in combination with concentrate-rich diets. Our study demonstrates that feeding a live yeast cells product, registered as a feed additive for improving zootechnical performance, had no impact on the toxicokinetics of OA under the chosen conditions. PMID- 19680880 TI - Determination of sterigmatocystin in cheese by high-performance liquid chromatography-tandem mass spectrometry. AB - Different cheese samples produced in Latvia (eight) and Belgium (13) were analysed for their sterigmatocystin (STC) content. Only two (9.5%) of the samples were positive for STC with concentration levels of 1.23 and 0.52 microg kg(-1), respectively. Five (24%) samples contained STC above the limit of detection (0.03 microg kg(-1)) but below the limit of quantification (0.1 microg kg(-1)), A sensitive liquid chromatography-electrospray positive ionisation-tandem mass spectrometry (LC-MS/MS) method, which was previously developed for the analysis of STC in grains, was modified and applied to the analysis of STC in cheese. This method involved sample extraction with acetonitrile-water (90 : 10, v/v), defatting with n-hexane, solid-phase extraction, separation on a reversed-phase C(18) column, and STC detection by LC-MS/MS. PMID- 19680881 TI - Determination of phthalates in raw bovine milk by gas chromatography/time-of flight mass spectrometry (GC/TOF-MS) and dietary intakes. AB - Low levels of phthalates, including di(2-ethylhexyl) phthalate (DEHP), in raw bovine milk were determined using gas chromatography/time-of-flight mass spectrometry (GC/TOF-MS). A fast and convenient process of sample treatment combined with TOF-MS analysis (medium resolution >5000), yielded good recoveries (85-125%) and low limits of detection (<0.002 mg kg(-1)). The most commonly used phthalate, DEHP, was found in 15 out of 30 samples monitored in this study. DEHP concentrations in raw milk ranged from not detected to 0.154 mg kg(-1), and the mean concentration was 0.057 mg kg(-1). The dietary intake of DEHP was about 0.004 mg kg(-1) body weight day(-1) if a child (24 months, 13 kg body weight) drinks 1 L day(-1) of milk that contains the mean concentration of DEHP found in raw milk. The estimated dietary intake corresponded to 8% of the European Union tolerable daily intake (TDI) of 0.05 mg kg(-1) body weight day(-1). Dimethyl phthalate (DMP) and di-n-butyl phthalate (DBP) were found from two and 20 samples, respectively, at low levels. Diethyl phthalate (DEP), butylbenzyl phthalate (BBP), and di-n-octyl phthalate (DnOP) were not found in any of the samples. PMID- 19680882 TI - A sensitive method for the determination of chlorine-36 in foods using accelerator mass spectrometry. AB - A method using accelerator mass spectrometry (AMS) has been developed to offer a more sensitive alternative to scintillation techniques for the determination of chlorine-36 ((36)Cl) in foods. The main problem in method development was the potential interference of the sulfur-36 ((36)S) isobar. This was overcome by reducing the sulfur level of acid digests of food by precipitation of chloride as silver chloride, then purification by washing, dissolution and reprecipitation to present silver chloride as the AMS target. The limit of detection was around 0.1 Bq kg(-1) and the limit of quantitation was around 0.2 Bq kg(-1). The AMS method was only semi-quantitative at the lowest levels of interest. To test the method a few samples of milk (five) and blackberries (three) collected from near two nuclear power stations as potential sources of contamination were analysed. Blackberries spiked at 0.2 Bq kg(-1) and milk spiked at 0.1 Bq kg(-1) could be distinguished from method blanks. There was no (36)Cl detectable in the unspiked samples. PMID- 19680883 TI - Isolation and identification of hydroxythiohomosildenafil in herbal dietary supplements sold as sexual performance enhancement products. AB - An unknown compound is detected and isolated from two herbal dietary supplements bought on the internet. The structure of the unknown compound is elucidated using ESI-MS/MS, NMR, UV and IR. The compound, named hydroxythiohomosildenafil, is identified as an analogue of sildenafil in which the oxygen atom is substituted with a sulfur atom in the pyrazolopyrimidine moiety, and a hydroxyethyl group instead of a methyl group is attached to the piperazinyl nitrogen. It is the first report of this compound being detected in herbal dietary supplements. The UV, IR and completely assigned NMR data of hydroxythiohomosildenafil is recorded. PMID- 19680884 TI - Effect of ion supplementation of a commercial soft drink on tooth enamel erosion. AB - Acidic soft drinks are potentially erosive for dental hard tissues. This in vitro study evaluated the effect of calcium, fluoride, iron and phosphate, supplemented alone or in combination to a commercial citric acid-based carbonated beverage on dental erosion. Ninety enamel samples (4 x 4 x 3 mm) were randomly allocated to nine groups (n = 10): G1-pure beverage (control); G2-with 1 mM Ca; G3-with 0.047 mM F; G4-with 1 mM Fe; G5-with 1 mM P; G6 - with 1 mM Ca and 0.047 mM F; G7-with 1 mM Ca and 1 mM P; G8-with 1 mM Fe and 0.047 mM F; and G9-with 1 mM Ca, 1 mM P, 0.047 mM F and 1.0 mM Fe. The samples were subjected to six pH cycles over a 24-h period. In each cycle, the samples were immersed in pure or modified beverage (1 min) and in artificial saliva (59 min). During the remaining period (18 h), the samples were maintained in artificial saliva. Enamel loss was assessed by profilometry (microm). Data were tested using ANOVA and Tukey's tests (p < 0.05). Highest enamel losses were observed in the control group (G1) and in the groups containing Fe (G4 and G8). The groups containing Ca (G2 and G6) showed significantly less wear compared to control. In conclusion, the modification of an erosive soft drink with low concentrations of Ca with or without F may reduced its erosive potential. PMID- 19680885 TI - Fate of imidacloprid and acetamiprid residues during black tea manufacture and transfer into tea infusion. AB - The objective of this paper is to investigate the loss/stability of neonicotinoids (imidacloprid and acetamiprid) residues during the manufacture of black tea and to study the transfer of these pesticides from made tea to its infusion. For orthodox black tea, the manufacturing process involves leaf harvesting (plucking two leaves and a bud), withering, rolling, fermentation (oxidation), and drying. Initial withering and final drying resulted in the loss of pesticide residues, but no significant reduction in residue levels resulted from the rolling and fermentation steps. The drying process resulted in a residue transfer of 64-70% and 69-74% of the initial level, whereas the brewing process resulted in a residue transfer of 37-39% and 45-49% of imidacloprid and acetamiprid, respectively, from dried tea into the infusion. The decrease in acetamiprid levels during drying was also significant (8-13%), whereas the total loss during the manufacturing process ranged from 26 to 31%. Further, brewing for longer periods (an extended brewing time) resulted in higher transfer (up to 44% for imidacloprid and 53% for acetamiprid) of pesticides to tea infusion. Further, the extent of pesticide leaching depends on its water solubility, partition coefficient, and the brewing time. PMID- 19680886 TI - Fate and distribution of pyrimethanil, metalaxyl, dichlofluanid and penconazol fungicides from treated grapes intended for winemaking. AB - Tempranillo grapes were immersed in solutions of pyrimethanil, metalaxyl, dichlofluanid and penconazol fungicides at different concentrations for several different times. Determinations of the fungicide residues was carried out by GC NPD, with an additional confirmation by GC-MS. The percentage absorption was determined as well as the distribution between surface, skin and pulp of the grapes. The percentage absorption ranged from 7.2 to 85.5%. These values depended on the time the grapes were in contact with the fungicide solutions. Residues were mainly found in skins, with percentages in skins ranging from 66.3 to 90.3% of total residues. In pulp, these values ranged from 3.5 to 31.0%. The overall methodology was applied to the determination of pyrimethanil in Tempranillo grapes treated with the recommended doses, respecting the safety period. Pyrimethanil residues found in treated grapes were higher in the skin. In grapes collected within the safety period (21 days), the levels found for fungicide residues were below maximum residues levels. PMID- 19680887 TI - Nitrate content of lettuce (Lactuca sativa L.) after fertilization with sewage sludge and irrigation with treated wastewater. AB - A romaine-type lettuce (Lactuca sativa L.) was cultivated over three crop seasons (spring 2005, spring 2006 and autumn-winter 2006) in six 36 m(2) plots in Alcazar de San Juan, Spain. A drip irrigation system was used to water all plots: five plots with drinking water and one plot with wastewater from the activated sludge system of a wastewater treatment plant (WWTP). One drinking water-irrigated plot was not fertilized (control). Five different treatments were applied to the soil: three organic mixtures (sewage sludge, sewage sludge mixed with pine bark and municipal solid waste with composted sludge) and a conventional fertilizer were applied to the four plots irrigated with drinking water. The last plot was irrigated with treated wastewater. The treatments were tested for their effect on plant growth and nitrate concentration in vegetable tissue. An increase in fresh weight in the lettuce was linked to the dosage of sewage sludge. The highest nitrate level was observed in the sewage sludge treatment in all crops and seasons, although, in general, all values were below the maximum limits established by the European Commission for nitrate content in fresh romaine lettuce. In the third crop season, a significant increase in nitrate content was observed in lettuce from organic treatments. Nitrate concentration in lettuce from irrigated treated wastewater was higher than control, although significant differences were not found. PMID- 19680888 TI - Simultaneous detection of the main black aspergilli responsible for ochratoxin A (OTA) contamination in grapes by multiplex real-time polymerase chain reaction. AB - This paper reports a duplex real-time polymerase chain reaction (PCR) assay for the simultaneous detection of members of the Aspergillus niger aggregate and A. carbonarius, which are the main responsible species for ochratoxin A (OTA) contamination in grapes. This single tube reaction targets the beta-ketosynthase and the acyl transferase domains of the poliketide synthase of A. carbonarius and the A. niger aggregate, respectively. Melting curve analysis using a SYBR Green I real-time PCR approach showed characteristic T(m)-values demonstrating the specific, efficient and balanced amplification of the two PCR fragments. Subsequently, a TaqMan real-time PCR approach was settled, using 6-carboxy fluorescein group (FAM) and VIC-labelled specific probes for automated detection. Results indicated no differences in sensitivity when using either the two sets of primers and probes in separate or in the same reaction. However, when both targets are in very different amounts, there is a preferential amplification of the target which is in more concentration. CT-values obtained in the presence of grape DNA were very similar to those observed when only fungal purified DNA was present, indicating that the grape DNA does not interfere in the real-time PCR reaction. This procedure provides a fast and accurate tool to monitor, in a single reaction, the presence of OTA-producing species in grapes which, to some extent, will facilitate OTA contamination surveys to guarantee food safety in the wine industry. PMID- 19680890 TI - Single-laboratory validation of a method for ergosterol determination in cereals. AB - Ergosterol (ERG) is a sterol produced by most fungi, but not by most plants. Thus, measurement of ERG in cereals makes it possible to determine the presence of fungi in cereals that can cause quality problems, such as mycotoxin contamination. This study developed and performed a single-laboratory validation for a method to test for ERG in various cereals. ERG was extracted by refluxing samples for 1 h with methanol-sodium hydroxide. ERG was extracted from the extract with hexane and then purified using a silica gel cartridge column. ERG was then separated and detected by reverse-phase high-performance liquid chromatography (HPLC). 'Within-day' recoveries of ERG at low levels were 92-99% with relative standard deviations (RSDs) of 3.2-6.5%. 'Between-day' recoveries of ERG at low levels were 97% and RSDs were 4.2-10.2%, respectively. Average recoveries of ERG over the range from 1.0 to 100.0 mg kg(-1) were 81-105% and RSDs were 3.9-16.3%. PMID- 19680889 TI - Occurrence of total aflatoxins, ochratoxin A and deoxynivalenol in foodstuffs available on the Lebanese market and their impact on dietary exposure of children and teenagers in Beirut. AB - Mycotoxins are fungal metabolites commonly present in food and constituting a potential threat to food safety. Total aflatoxins (AFT), ochratoxin A (OTA) and deoxynivalenol (DON) are among the most widespread mycotoxins. We assessed the occurrence of AFT, OTA and DON in some foodstuffs available on the Lebanese market and evaluated the potential risk to the health of children and teenagers in Beirut from dietary exposure to these mycotoxins. Analytical data on the contamination of raw and processed cereals and cereal-based products, pulses and nuts were collected. The mean levels of AFT, OTA and DON were calculated for each food items. Levels of dietary exposure were obtained by combining food consumption data with the mean mycotoxin levels, adopting a deterministic approach. Intakes were calculated for average and high consumers (75th and 95th percentile) among children and teenagers, and compared with the provisional tolerable daily intake (PTDI) or provisional tolerable weekly intake (PTWI) of the respective mycotoxin. The results showed that mycotoxin levels in the food samples were generally below national/European maximum limits. However, high levels of AFT, OTA and DON were found in some samples of nuts, biscuits and bread. The calculated intake for AFT exceeded its respective PTDI in all groups by a factor ranging from 3 to 7. The intakes of OTA and DON were found to be below the threshold of toxicological concern established for these mycotoxins by international expert groups, although the intake of DON in children at the highest percentile (P95) was close to its PTDI. Our study highlights the need to reconsider the maximum permissible levels of total aflatoxins in some foodstuffs and to establish regulatory guidelines regarding OTA and DON. The results also emphasise the need for routine monitoring of the levels of mycotoxin contamination in foodstuffs that are highly consumed by Lebanese children and teenagers, i.e. cereals and cereal-based products. PMID- 19680892 TI - Assessment of a new lateral flow immunochromatographic (LFIC) assay for the okadaic acid group of toxins using naturally contaminated bivalve shellfish from the Portuguese coast. AB - A new rapid assay for the okadaic acid group of toxins, based on lateral flow immunochromatographic (LFIC) test strips developed by Jellett Rapid Testing Ltd., was assessed on naturally contaminated bivalves from the Portuguese coast. One prototype was evaluated using samples harvested during 2005, extracted with 80% methanol, followed by dilution with the running buffer of a methanolic extract after alkaline hydrolysis for esters. The second prototype was assessed using samples harvested during 2006, extracted with 100% methanol and, after alkaline hydrolysis, the methanol was evaporated by a nitrogen stream followed by re suspension with the running buffer. The first prototype failed to detect 20% of samples that were positive by LC-MS in the range 160-480 microg kg(-1), and were classified as negative or trace level by LFIC. The presence of methanol in the extracts made correct detection of toxins more difficult. The second prototype classified as positive all samples above 160 microg kg(-1), as confirmed by LC MS. However, in the second prototype, matrix effects were a major drawback and led to 45% false positives, particularly for mussels, due to compounds in shellfish extracts interfering with the antibodies and reducing the test line intensity. Extraction with a higher percentage of methanol was thought responsible for these matrix effects. Regarding sample migration, both prototypes needed one hour before reading. In an attempt to speed-up sample preparation, a direct digestion of bivalve tissues with sodium hydroxide was evaluated. Low recoveries for esters were found by LC-MS with this hydrolysis technique compared to conventional hydrolysis of methanolic extracts. While prototype A was not sensitive enough, prototype B had too many false positives to be of use to the shellfish industry or in a monitoring program. PMID- 19680893 TI - Determination of yessotoxins and pectenotoxins in shellfish by capillary electrophoresis-electrospray ionization-mass spectrometry. AB - Conditions for the determination of lipophilic marine toxins, such as yessotoxins and pectenotoxins (PTX)-6, were investigated with capillary electrophoresis coupled to mass spectrometry (MS) with an electrospray ionization source. After optimization, a simple and MS compatible alkaline volatile buffer solution of ammonium acetate was selected as background electrolyte, with isopropanol/water (80/20, v/v) sheath liquid modified with ammonium acetate used at the electrospray ionization (ESI) source. Previously to capillary electrophoresis (CE) separations, the application of an on-line sample pre-concentration approach based on field-amplified sample stacking was accomplished to increase sensitivity. As a result, the limits of detection provided by capillary electrophoresis-electrospray ionization-mass spectrometry (CE-ESI-MS) were 0.02 microg ml(-1) (0.01 microg g(-1)), which corresponded to 1.25 pg for yessotoxin and 0.25 microg ml(-1) (0.13 microg g(-1) and 13.25 pg on capillary) for PTX-6. Accuracy tests showed 97.7% recovery from spiked blank mussel samples that showed no significant matrix influence running under optimal conditions. Intermediate precision was close to 4% relative standard deviation (RSD) for the migration time, and an RSD of 7.5% for peak areas. The method was successfully applied to naturally contaminated seafood samples in which yessotoxins and pectenotoxins-6 were clearly determined. This work demonstrated the potential of CE-ESI-MS to be applied for a sensitive determination of lipophilic toxins from the marine environment as alternative to liquid chromatography-electrospray ionization single quadrupole mass spectrometry (LC-ESI-MS) for this purpose. PMID- 19680891 TI - Preparation and characterization of the conjugated Fusarium mycotoxins zearalenone-4O-beta-D-glucopyranoside, alpha-zearalenol-4O-beta-D-glucopyranoside and beta-zearalenol-4O-beta-D-glucopyranoside by MS/MS and two-dimensional NMR. AB - Glucosides of several Fusarium mycotoxins occur in naturally infected cereals and may contribute to an increased content to the total mycotoxin load of food and feed. The paper presents the results of a fermentation procedure to produce zearalenone-4O-beta-D-glucopyranoside from zearalenone using an engineered Saccharomyces cerevisiae strain, expressing the Arabidopsis thaliana UDP glucosyltransferase UGT73C6. About 24 mg of zearalenone-4O-beta-D-glucopyranoside was obtained from 50 mg of zearalenone and further purified. A total of 10 mg of the glucoside were reduced with sodium borohydride, yielding 4.1 mg alpha zearalenol-4O-beta-D-glucopyranoside and 4.5 mg beta-zearalenol-4O-beta-D glucopyranoside at purities higher than 99%. To confirm the identities of the three produced glucosides, MS and MS/MS spectra were acquired using negative electrospray ionization. Besides the deprotonated ions at m/z 479 or 481, respectively, in full-scan mode, fragments, adducts, and dimers were recorded and assigned. MS/MS spectra of the glucosylated substances yielded the deprotonated ions of the mycotoxins zearalenone, alpha-zearalenol, beta-zearalenol and their fragments, respectively. Unambiguous structural assignment of the three substances was achieved using two-dimensional NMR methods. This way, the glucose attachment to position C-4, the beta-configuration of the sugar unit and the stereo-chemical assignment of the zearalenol hydroxyl group at C-6' were proven. PMID- 19680894 TI - Extraction recoveries and stability of diarrhetic shellfish poisoning (DSP) toxins in naturally contaminated samples. AB - During the last few years the occurrence of a high percentage of esters of diarrhetic shellfish poisoning (DSP) toxins has been observed in shellfish from the Portuguese coast. Most of the commercial bivalves contain DSP toxins in ester forms, either acyl derivatives of okadaic acid (OA) or of dinophysistoxin-2 (DTX 2). The stability of these toxins in shellfish tissues and in raw methanol extracts was investigated in two different naturally contaminated species, mussel and carpet shell, over a 4-week period. The results for both species revealed that DSP toxins were more stable in tissue than in raw methanol extracts. Losses of DSP toxins were seen in the first 2 weeks and were more than 30%, but after that a period of stabilization was observed. The decrease was due probably from losses of esters of OA and DTX-2, the free toxins were stable over the period studied. The extraction most commonly used for chemical and biochemical assays relied on methanolic extraction with aqueous 80% methanol. In this work we have tested the extraction solvent on the extractability of DSP toxins from several naturally contaminated species. A single dispersive extraction with methanol, with solvent ratios of 70%, 80%, 90% and 100%, were tested. After alkaline hydrolysis of esterified toxins and clean-up with hexane and dichloromethane, the samples were analysed by liquid chromatography-mass spectrometry (LC-MS). The recovery of DSP toxins increased with increasing percentages of methanol up to 90%. A decrease in recovery with 100% methanol was observed probably due to problems during the liquid-liquid partitioning. PMID- 19680895 TI - Solid-phase extraction clean-up of ciguatoxin-contaminated coral fish extracts for use in the mouse bioassay. AB - Florisil solid-phase extraction (SPE) cartridges were used for purifying ciguatoxin (CTX)-contaminated coral fish extracts, with the aim of removing extracted lipid but retaining optimal level of CTXs in the purified fractions. The CTX-containing fraction (target fraction) in fish ether extract was isolated and purified by eluting through a commercially available Florisil cartridge with hexane-acetone-methanol solvent mixtures of increasing polarity (hexane-acetone (4:1, v/v) < acetone-methanol (7:3, v/v) < 100% methanol). Application of Florisil SPE using acetone-methanol (7:3, v/v) condition facilitated the separation of 4.2 +/- 0.4 mg (mean +/- standard error of the mean (SEM)) of purified target fraction from 20 mg ether extract with good retention of CTXs. The mouse bioassay was used to demonstrate that the average CTX recovery of the target fraction from CTX-spiked samples was 75.8% +/- 3.3%, which was significantly increased by 96.7% +/- 15% when compared with CTX recovery from ether extracts (44.8% +/- 5.2%) without performing SPE purification. Over 70% of non-target lipids were removed in which no CTX toxicity was found. Moreover, the target fractions of both CTX-spiked and naturally CTX-contaminated samples gave more prominent toxic responses of hypothermia and/or induced more rapid death of the mice. The use of acetone-methanol (7:3, v/v) condition in the elution could significantly improve overall recovery of CTXs, while minimizing the possible interferences of lipid matrix from co-extractants on mice. PMID- 19680896 TI - Photocatalytic properties of nanocrystalline titanium dioxide films in the degradation of domoic acid in aqueous solution: potential for use in molluscan shellfish biotoxin depuration facilities. AB - Domoic acid (DA) is a water-soluble marine neurotoxin produced and released by certain species of the diatom genus Pseudo-nitzschia. Present in coastal waters, it can be a threat to public health and marine life, and can result in severe economic losses to the molluscan shellfish and crustacean harvesting industries. Here we report on the efficiency of nanocrystalline (NC) titania (TiO(2)) thin films used as a photocatalyst in the ultraviolet light photodegradation of DA. Titanium dioxide thin films produced by a sol-gel dip-coating method in the presence of polyethylene glycol of different molecular weights (200, 400 and 600) were deposited on glass substrates and crystallised at 90 degrees C. The films were characterised using spectroscopic ellipsometry, Fourier transform infrared spectrometer (FTIR), scanning electron microscopy (SEM), X-ray diffraction (XRD) and Raman spectroscopy. The photocatalytic activity measurements were carried out by immersing the NC TiO(2) films in a DA solution (2500 ng ml(-1)) and then exposing them for various times at room temperature to UVA irradiation (lambda = approximately 350 nm). The degradation of DA, quantified by HPLC analysis, was not significant when using daylight or ultraviolet light irradiation alone, whereas the NC TiO(2) films prepared at low temperature proved to be a very efficient photocatalyst when used in conjunction with UVA light. The effectiveness of the photodegradation was improved by increasing molecular weight of polyethylene glycol, which increased the thickness of the film. The presence, transformation and degradation of three DA isomers were observed. The approach may eventually be practical for destroying DA in seawater used by aquaculture industry depuration facilities. PMID- 19680897 TI - Pentachlorophenol, polychlorodibenzodioxin and polychlorodibenzofuran in eggs from hens exposed to contaminated wood shavings. AB - Laying hens may be exposed to pentachlorophenol (PCP) present in bedding materials derived from treated timber. As a result, this chemical and its contaminants or degradation products, such as polychlorodibenzodioxins (PCDDs) and polychlorodibenzofurans (PCDFs), may be present in eggs. The litter-to-eggs transfer and depletion of these compounds were studied in a flock of laying hens reared on contaminated wood shavings. PCP determination was carried out via high resolution gas chromatography coupled to low resolution mass spectrometry/mass spectrometry (HRGC-LRMS/MS); PCDDs and PCDFs were quantified by HRGC-HRMS (SIM). After substitution of the litter contaminated with PCDDs and PCDFs at an average of 50 pg WHO-TE g(-1) and with PCP at 15 microg g(-1), pooled eggs from six selected hens were sampled twice a month for the depletion study. At steady state, PCDDs and PCDFs showed a transfer ratio of 0.9; for PCP it was 0.03. PCP concentration in eggs (500 ng g(-1) whole weight) fell sharply the second week after exposure withdrawal; for PCDDs and PCDFs (47 pg WHO-TE g(-1) fat, at day 0), the overall TEQ half-life was estimated at an average of 3.8 weeks. Due to differences in toxicokinetics (a faster depletion), PCP does not appear to be a suitable marker of PCDD and PCDF violative levels. However, the prominent analytical contribution of H(7)CCD and O(8)CCD in the contamination profile may help to trace the source of contamination. Among congeners, 2,3,7,8 T(4)CDF exhibited a different depletion pattern, indicating a possible mechanism of active transport. PMID- 19680898 TI - Determination of bisphenol A in canned fatty foods by coacervative microextraction, liquid chromatography and fluorimetry. AB - Decanoic acid reverse micelle-based coacervates were used to provide simple, rapid and almost solventless extraction of bisphenol A (BPA) from canned fatty foods. The procedure involved the extraction of 200-400 mg of homogenised food sample with an aqueous solution containing 20% THF and 200 mg of decanoic acid, conditions under which the coacervate (around 550 microl) formed in situ and instantaneously. The overall sample treatment took about 30 min and several samples could be simultaneously treated using conventional laboratory equipment. No clean-up or solvent evaporation were required before determination of BPA by liquid chromatography and fluorescence detection. Recoveries in samples were between 90 and 99%, with relative standard deviations in the range 2-7%. The limit of quantification ranged 29-15 ng g(-1) for 200-400 mg of sample, being far below the current specific migration limit (SML) set by the European Commission (600 ng g(-1)). The method was successfully applied to the determination of BPA in the solid content of canned fish (from 20 to 129 ng g(-1)) and meat (from undetected to 37 ng g(-1)). PMID- 19680900 TI - Characterisation of vitamin B12 immunoaffinity columns and method development for determination of vitamin B12 in a range of foods, juices and pharmaceutical products using immunoaffinity clean-up and high performance liquid chromatography with UV detection. AB - New rapid and simpler procedures, using immunoaffinity columns, have been developed for the determination of vitamin B12 in a range of samples including three different US National Institute of Standard and Technology (NIST) Reference Materials, infant formula, powdered energy drinks and bars, wheat breakfast cereal, carbonated soft drinks, fruit juices and vitamin B12 tablets. The procedures involved extraction of vitamin B12 using water or sodium acetate buffer and enzyme digestion (using pepsin or alpha-amylase, or both) if necessary. The extract was clarified and passed through "EASI-EXTRACT Vitamin B12", an immunoaffinity column containing monoclonal antibody with high affinity and specificity to vitamin B12. Subsequently, the vitamin B12 immunoaffinity column was washed with 10 ml water and the vitamin B12 was released from the column with 3 ml methanol. Following evaporation, the samples were reconstituted in mobile phase and analysed by HPLC-UV at 361 nm on an ACE 3AQ analytical column using a gradient elution consisting of 0.025% trifluoroacetic acid in water and acetonitrile. Analysis of three types of NIST Standard Reference Materials in triplicate demonstrated the results of the immunoaffinity column method were comparable to microbiological assay results. Method repeatability was determined for all samples analysed and ranged between 0.8 and 10%, demonstrating the method was repeatable with complex matrices (NIST 2383) containing low levels of vitamin B12 (0.44 microg per 100 g), as well as simpler matrices, such as vitamin tablets containing high levels (2000 microg per 0.849 g) of vitamin B12. PMID- 19680899 TI - Caffeine levels in beverages from Argentina's market: application to caffeine dietary intake assessment. AB - The caffeine content of different beverages from Argentina's market was measured. Several brands of coffees, teas, mates, chocolate milks, soft and energy drinks were analysed by high-performance liquid chromatography (HPLC) with ultraviolet detection. The highest concentration level was found in short coffee (1.38 mg ml( 1)) and the highest amount per serving was found in instant coffee (95 mg per serving). A consumption study was also carried out among 471 people from 2 to 93 years of age to evaluate caffeine total dietary intake by age and to identify the sources of caffeine intake. The mean caffeine intake among adults was 288 mg day( 1) and mate was the main contributor to that intake. The mean caffeine intake among children of 10 years of age and under was 35 mg day(-1) and soft drinks were the major contributors to that intake. Children between 11 and 15 years old and teenagers (between 16 and 20 years) had caffeine mean intakes of 120 and 240 mg day(-1), respectively, and mate was the major contributor to those intakes. Drinking mate is a deep-rooted habit among Argentine people and it might be the reason for their elevated caffeine mean daily intake. PMID- 19680901 TI - Dietary exposure to acrylamide from potato crisps to the Spanish population. AB - Potato crisps are one of the food commodities that contribute most to overall dietary human exposure of acrylamide. This investigation has estimated the dietary exposure to acrylamide form potato crisps in the Spanish population. Sampling of potato crisps (n = 36) from 16 different producers were carried out in March 2008. An average level of 740 microg kg(-1) (ranging from 81 to 2622 microg kg(-1); minimum to maximum) and a median of 592 microg kg(-1) were obtained. Acrylamide levels in marketed potato crisps have been significantly reduced (nearly to 50%) compared with a previous sampling performed 4 years earlier. The observed signal value (90th percentile) was 1377 microg kg(-1) with 86% of the samples with acrylamide levels lower than 1000 microg kg(-1). Dietary exposure to acrylamide from potato crisp consumption in the total Spanish population was estimated to be 0.042 microg kg(-1) body weight day(-1) by using a deterministic approach based on the National consumption database. In a second study, dietary exposure (based on a 3-day food record) was determined to be 0.053 microg kg(-1) body weight day(-1) for the adult population (17-60 years) and 0.142 microg kg(-1) body weight day(-1) for children (7-12 years). The contribution of potato crisps to the estimated dietary acrylamide exposure of the Spanish population is moderate as compared with other European Member States. PMID- 19680902 TI - Analysis of heterocyclic amines and beta-carbolines by liquid chromatography-mass spectrometry in cooked meats commonly consumed in Korea. AB - Heterocyclic amines (HAs), which form in meats during heating and cooking, are recognized as mutagenic and carcinogenic compounds. In this study, 13 HAs and 2 beta-carbolines (BCs) were analyzed in cooked Korean meat products, including griddled bacon, griddled pork loin, boiled pork loin, boiled chicken meat, chicken meat stock, chicken breast for salad and chicken patty. The samples were either cooked in the laboratory or purchased from local fast-food restaurants. The HAs and BCs in the samples were separated using solid-phase extraction and were analyzed by high performance liquid chromatography-mass spectrometry (HPLC MS). The most frequently detected HAs and BCs in the cooked meats were harman (1 methyl-9H pyrido[4,3-b]indole; 990.9 ng g(-1)), norharman (9H-pyrido[4,3 b]indole; 412.7 ng g(-1)) and PhIP (2-amino-1-methyl-6-phenylimidazo[4,5 b]pyridine; 258.2 ng g(-1)). The griddled pork loin and bacon contained higher levels of norharman, harman and PhIP than the other cooked meats. PhIP, which is classified as a Group 2B carcinogen by the International Agency for Research on Cancer, had levels of 258.2 and 168.2 ng g(-1) in the griddled pork loin and griddled bacon, respectively. The griddled bacon was the only sample containing TriMeIQx (2-amino-3,4,7,8-tetramethylimidazo[4,5-f]quinoxaline; 79.9 ng g(-1)). IQ (2-amino-3-methyl imidazo[4,5-f]quinoline), 7,8-DiMeIQx (2-amino-3,7,8 trimethylimidazo[4,5-f]quinoxaline), 4,8-DiMeIQx (2-amino-3,4,8 trimethylimidazo[4,5-f]quinoxaline) and AalphaC (2-amino-9H-pyrido[2,3-b]indole) were detected at trace levels in all samples. PMID- 19680903 TI - Rapid determination of 19 quinolone residues in spiked fish and pig muscle by high-performance liquid chromatography (HPLC) tandem mass spectrometry. AB - A multiresidue method has been developed for the confirmation and quantification of 19 quinolones (enrofloxacin, ciprofloxacin, norfloxacin, ofloxacin, flumequine, oxolinic acid, difloxacin, sarafloxacin, sparfloxacin, danofloxacin, fleroxacin, marbofloxacin, enoxacin, orbifloxacin, pefloxacin, nalidixic acid, pipemidic acid, lomefloxacin and cinoacin) in pig and fish by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The samples were extracted with acetonitrile, analytes separated by LC on a C18 column using 0.1% formic acid-methanol with a linear gradient elution programme, and detected by MS/MS. The linear range was 0.3-50 microg kg(-1) with correlation coefficients (r) more than 0.9956. The limits of detection were 0.1 microg kg(-1). Mean recoveries for each analyte in pig muscle and fish ranged from 75.3% to 96.3% and from 79.7% to 94.2% with relative standard deviations below 10%. The method is fast, safe, sensitive and precise, and can be used simultaneously to analyse residual quinolones. PMID- 19680904 TI - Development of immunoassays for the detection of sulfamethazine in swine urine. AB - The use of sulfonamides, such as sulfamethazine (SM2), in pig production is recognized as a public health risk as it inevitably results in sulfamethazine residues in pork. This study is aimed at establishing rapid, simple, reliable methods, with both sensitivity and specificity, for detecting sulfamethazine residues. For this purpose, monoclonal antibodies against sulfamethazine were prepared and characterized. No cross-reaction of the monoclonal antibodies was identified with other sulfonamides or analytes. Based on the competitive immunoassay principle, an indirect competitive ELISA kit (SM2 kit) and a rapid detection strip for detecting sulfamethazine residues were developed using monoclonal antibodies and the colloidal gold technique. The indirect competitive ELISA kit and the strip assay could be performed within 2 h and 5-10 min, respectively. The results showed that the detection limits were 1 ng ml(-1) for the indirect competitive ELISA kit and 8 ng ml(-1) with the unaided eye and 1 ng ml(-1) with the strip reader for the rapid strip assay. Comparing the HPLC method with the SM2-kit or the strip in pig urine spiked with standards of SM2, the difference was <4.6% for SM2-kit and 4.3% for the strip. The two methods are suitable for the rapid screening of sulfamethazine residues in swine urine. Experimental data revealed that the two methods, especially the strip, proved to be sensitive, specific, rapid and easy to use for the quantitative, semi quantitative or qualitative detection of SM2 residues in swine urine. PMID- 19680905 TI - Accumulation of trace elements in cattle from rural and industrial areas in Belgium. AB - Trace element concentrations were determined in meat, kidney and liver tissue of cattle, which had resided more than 18 months in areas historically contaminated by emissions from the non-ferrous metal industry or in regions with normal background metal contamination levels in Belgium. Trace element levels were determined by inductively coupled plasma-mass spectrometry. Lead, arsenic and cadmium concentrations in meat were low for all cattle. Concentrations in kidneys were, respectively, 1.8-, 2.2- and 2.5-fold higher in cattle from the contaminated areas compared to animals from reference areas. The European maximum level for cadmium in kidney was exceeded by 75% of the kidneys from contaminated areas and 47% of the kidneys from reference areas. Lead, arsenic and cadmium concentrations in bovine livers from contaminated areas were increased by a factor of 2.3. Zinc levels in livers and kidneys were within normal ranges, but 20% higher in the contaminated areas compared to reference areas. Copper levels in livers were high in all areas. Dairy cattle accumulated significantly higher levels of cadmium in kidneys and of lead in liver and kidneys than did beef cattle. PMID- 19680906 TI - Pesticide multiresidue analysis of peanuts using automated gel permeation chromatography clean-up/gas chromatography-mass spectrometry. AB - A method based on automated gel permeation chromatography (GPC) and on-line quantitative concentration was developed for the determination of residues of 38 typically used pesticides in high-oil peanuts. Pesticides were extracted using acetonitrile into an oil fraction containing little of the peanut matrix. After changing the solvent to the mobile phase (1 : 1 cyclohexane : ethyl acetate), clean-up was carried out using GPC, the final collected solution being automatically concentrated to a fixed volume and transferred into vials for gas chromatography (GC) injection. The pesticides were analysed by gas chromatography mass spectrometry (GC/MS) in the selected ion monitoring (SIM) mode. Average recoveries (spiked levels of 0.02, 0.05, and 0.1 microg g(-1)) were between 70 and 117%. The relative standard deviation of the method was 3.5-21% (n = 6). This approach gave comparable results with a previously published method. PMID- 19680907 TI - Organophosphorus pesticide residues in milled rice (Oryza sativa) on the Chinese market and dietary risk assessment. AB - The present study investigates the occurrence of acetylcholinesterase (AChE) inhibiting organophosphorus (OP) pesticide residues in milled rice samples obtained form local markets in China during the period 2004-2006 and estimates their cumulative exposure. Concentrations of OP pesticides were determined by gas chromatography with flame photometric detection (GC-FPD). The results showed that 9.3% of the samples contained detectable residues of at least one of the seven target OP pesticides (chlorpyrifos, dichlorvos, omethoate, methamidophos, parathion-methyl, parathion and triazophos) mainly used for agriculture in China, with concentrations ranging 0.011-1.756 mg kg(-1). Rice consumption data was obtained from an individual food consumption survey. Relative potency factors (RPFs) for each pesticide were calculated with methamidophos as the index compound (IC), using 1- or 2-year chronic non-observed adverse effect levels (NOAEL) for AChE inhibition, mostly in rat brain, obtained from international evaluations of pesticides. Exposure to AChE-inhibiting pesticides for the population above 7 years old at P99.9 represented 52-94.5% of the acceptable daily intake (ADI) expressed as methamidophos. Estimated exposure for children aged 2-4 and 4-7 years at P99.9 were 119 and 104.3% of the ADI level, respectively. This study suggests that a yearly monitoring program for OP pesticide residues and strict implementation of the national safety standard for milled rice is necessary. PMID- 19680908 TI - Assessment of average exposure to organochlorine pesticides in southern Togo from water, maize (Zea mays) and cowpea (Vigna unguiculata). AB - Drinking water, cowpea and maize grains were sampled in some potentially exposed agro-ecological areas in Togo and analysed for their contamination by some common organochlorine pesticides. A total of 19 organochlorine pesticides were investigated in ten subsamples of maize, ten subsamples of cowpea and nine subsamples of drinking water. Analytical methods included solvent extraction of the pesticide residues and their subsequent quantification using gas chromatography-mass spectrometry (GC/MS). Estimated daily intakes (EDIs) of pesticides were also determined. Pesticides residues in drinking water (0.04-0.40 microg l(-1)) were higher than the maximum residue limit (MRL) (0.03 microg l( 1)) set by the World Health Organization (WHO). Dieldrin, endrin, heptachlor epoxide and endosulfan levels (13.16-98.79 microg kg(-1)) in cowpea grains exceeded MRLs applied in France (10-50 microg kg(-1)). Contaminants' levels in maize grains (0.53-65.70 microg kg(-1)) were below the MRLs (20-100 microg kg( 1)) set by the Food and Agriculture Organization (FAO) and the WHO. EDIs of the tested pesticides ranged from 0.02% to 162.07% of the acceptable daily intakes (ADIs). Population exposure levels of dieldrin and heptachlor epoxide were higher than the FAO/WHO standards. A comprehensive national monitoring programme on organochlorine pesticides should be undertaken to include such other relevant sources like meat, fish, eggs and milk. PMID- 19680909 TI - Endotoxin and biogenic amine levels in Atlantic mackerel (Scomber scombrus), sardine (Sardina pilchardus) and Mediterranean hake (Merluccius merluccius) stored at 22 degrees C. AB - Whole Atlantic mackerel (Scomber scombrus), sardine (Sardina pilchardus) and Mediterranean hake (Merluccius merluccius) from the Croatian Adriatic were stored at 22 degrees C and changes in histamine, putrescine, tyramine and cadaverine levels were monitored in relation to bacterial endotoxin. After 12 h, histamine levels in sardine were above the legal limit of 50 mg kg(-1), set by the US Food and Drug Administration, and an increase in putrescine content preceded the increase in histamine. After 24 h, histamine contents in mackerel and sardine reached 1090 +/- 101 and 577 +/- 275 mg kg(-1), respectively, which exceeded the toxic threshold of 500 mg kg(-1). At the same time, the putrescine content was also high in both fish (353-420 mg kg(-1)). The time-course of endotoxin production was similar in all fish species stored at 22 degrees C. A high correlation was found between endotoxin and histamine, and between endotoxin and putrescine in mackerel and sardine. On the other hand, high endotoxin levels in hake, after 24 h, were associated with the low histamine and putrescine content (40-60 mg kg(-1)). PMID- 19680910 TI - Occurrence of Fusarium mycotoxins in maize imported into the UK, 2004-2007. AB - This study examined a total of 82 consignments of French and Argentinean raw maize as received at maize mills in the UK between 2004 and 2007. Samples were analysed for deoxynivalenol (DON), nivalenol (NIV), other trichothecenes, zearalenone (ZON), and fumonisins B(1), B(2), and B(3) (FB(1), FB(2), and FB(3)) using fully validated analytical methods with limits of quantification of 10 microg kg(-1) for DON, NIV, and each fumonisin mycotoxin and 3 microg kg(-1) for ZON. All samples except two containing fumonisins met the European Commission statutory maximum permissible levels for DON, ZON, and FB(1) + FB(2) as operating in 2007. The maximum concentrations found for DON, NIV, ZON, and FB(1) + FB(2) were 444, 496, 165 and 5002 microg kg(-1), respectively. Fumonisins were detected in almost every sample with 65% of Argentinean maize containing more than 1000 microg kg(-1) of FB(1) + FB(2). In contrast, ZON was not detectable in almost 50% of consignments. During this period there was a distinct difference in mycotoxin concentrations between harvests and geographic origin. Flint maize from Argentina usually contained lower concentrations of DON and related trichothecenes and higher levels of fumonisins than maize from France, although concentrations of fumonisins up to 2000 microg kg(-1) or greater occurred in samples from both regions. The incidence and concentrations of fumonisins were similar to those in a similar previous survey, while zearalenone concentrations were lower. The distribution of mycotoxins in multi-hold ships was also investigated showing that fumonisins were much more evenly distributed than DON, thus indicating their general level in the ship as a whole. The effect of cleaning regimes was found to be very variable, especially for DON, ranging from no removal of mycotoxins to greater than 50% in some instances, but was not related to concentration. Evidence here suggests that while cleaning is essential for removing foreign bodies before milling, it cannot be used as a reliable tool for reducing mycotoxins. PMID- 19680911 TI - Distribution of aflatoxins and fumonisins in dry-milled maize fractions. AB - The aim was to evaluate the distribution of aflatoxins and fumonisins in fractions derived from the dry-milling of contaminated maize. Two maize lots with different contamination levels were processed and sampled: the first (maize 1) had aflatoxin B(1) (AFB(1)) and fumonisin B(1) (FB(1)) levels of 3.6 and 5379 microg kg(-1), respectively; the second (maize 2) had corresponding levels of 91.1 and 8841 microg kg(-1), respectively. The cleaning step reduced AFB(1) and FB(1) levels by 8 and 11% in maize 1 and by 57 and 34% in maize 2. The subsequent removal of bran and germ led to a further decrease in contamination levels in the products destined for human consumption. In the latter, AFB(1) was uniformly distributed, while FB(1) was concentrated in the finer size fractions. Contamination of raw maize 1 (3.6 microg kg(-1)) was below the European Union AFB(1) limit of 5 microg kg(-1) for unprocessed maize, but among the final products only coarse flour (1.7 microg kg(-1)) was within the European Union limit of 2 microg kg(-1), while grits and fine flour showed higher levels (2.7 and 2.5 microg kg(-1), respectively). As regards cleaned maize, a different distribution of the two toxins was observed in the kernels: AFB(1) contamination was more superficial and concentrated in germ, while FB(1) contamination affected the inner layers of the kernels. PMID- 19680912 TI - Delivery systems for biological control agents to manage aflatoxin contamination of pre-harvest maize. AB - While soil application of a competitive non-toxigenic Aspergillus flavus strains is successful in reducing aflatoxin contamination in certain crops, direct application to aerial reproductive structures could be more effective for maize. A sprayable, clay-based water-dispersible granule formulation was developed to deliver non-toxigenic A. flavus strain K49 directly to maize ears. The efficacy of the K49 water-dispersible granule in mitigating aflatoxin in maize (Zea mays L.) was evaluated. Field studies were conducted to compare K49 colonization and effectiveness in reducing aflatoxin contamination when applied either as a soil inoculant or as a directed spray in plots infested with toxigenic strain F3W4. Fifty percent of non-toxigenic A. flavus was recovered from non-treated controls and from plots soil inoculated with K49 on wheat. In spray treatments with formulated or unformulated K49 conidia, over 90% of A. flavus recovered was non toxigenic. Soil-applied K49 reduced aflatoxin contamination by 65% and spray applications reduced contamination by 97%. These findings suggest direct spray application of non-toxigenic A. flavus strains may be better than soil inoculation at controlling maize aflatoxin contamination and that a water dispersible granule is a viable delivery system for maintaining viability and efficacy of the biological control agent, K49. PMID- 19680913 TI - Dominance of Group 2 and fusaproliferin production by Fusarium subglutinans from Iowa maize. AB - Fusarium subglutinans (teleomorph Gibberella subglutinans, member of the Gibberella fujikuroi complex) is an important toxigenic pathogen of maize. Recently, two cryptic species (Groups 1 and 2) have been described within F. subglutinans, but little is known about the occurrence of the two groups in North America or their relative capacities to produce mycotoxins. In this study, 58 F. subglutinans strains from kernels of maize grown in Iowa, USA, were evaluated for cryptic speciation and production of the mycotoxins fusaproliferin and beauvericin. Fifty-six of the 58 strains (97%) belonged to Group 2, and two strains belonged to Group 1, based on restricted fragment length polymorphisms derived from amplification of histone H3 and beta-tubulin gene fragments. Fifty four Group 2 strains and both Group 1 strains produced fusaproliferin at concentrations ranging from 12 to 3000 microg g(-1) of solid maize culture. None of the F. subglutinans strains from Iowa produced beauvericin at detectable amounts, although most F. subglutinans strains from Europe and elsewhere are beauvericin producers. These results indicate that F. subglutinans strains infecting maize kernels in Iowa belong almost exclusively to Group 2 and that they have a high potential for fusaproliferin production; furthermore, the results confirm an association between Group 2 genotypes and lack of beauvericin production. This is the first report characterizing the phylogenetic groups of F. subglutinans occurring in Iowa; the predominance of Group 2 suggests that populations of the fungus in Iowa and Europe remain isolated from each other. Fusaproliferin contamination of grain appears to be a risk wherever F. subglutinans occurs, but beauvericin contamination from F. subglutinans is associated only with Group 1. PMID- 19680914 TI - Determination of the overall migration from silicone baking moulds into simulants and food using 1H-NMR techniques. AB - Different silicone baking moulds (37 samples) were characterized with respect to potential migrating substances using 1H-NMR, RP-HPLC-UV/ELSD and GC techniques. In all cases cyclic organosiloxane oligomers with the formula [Si(CH3)2-O]n were identified (n = 6 ... 50). Additionally, linear, partly hydroxyl-terminated organosiloxanes HO-[Si(CH3)2-O]n-H (n = 7 ... 20) were found in 13 samples. No substances other than siloxanes could be detected, meaning the migrants mainly consist of organopolysiloxanes. Based on this knowledge, a 1H-NMR quantification method for siloxanes was established for the analysis of both simulants and foodstuffs. Validation of the 1H-NMR method gave suitable performance characteristics: limit of detection 8.7 mg kg(-1) oil, coefficient of variation 7.8% (at a level of 1.0 mg kg(-1) food). Migration studies were carried out with simulants (olive oil, isooctane, ethanol (95%), Tenax) as well as preparation of different cakes. From the 1st to 10th experiment, siloxane migration into cakes only slightly decreased, with a significant dependence on fat content. Migration never exceeded a level of 21 mg kg(-1) (3 mg dm(-2)) and was, therefore, well below the overall migration limit of 60 mg kg(-1) (10 mg dm(-2)). However, migration behaviour into simulants differed completely from these results. PMID- 19680915 TI - Simultaneous detection of DNA from 10 food allergens by ligation-dependent probe amplification. AB - The simultaneous detection of DNA from different allergenic food ingredients by a ligation-dependent probe amplification (LPA) system is described. The approach allows detection of several targets in a one-tube assay. Synthetic oligonucleotides were designed to detect DNA from peanuts, cashews, pecans, pistachios, hazelnuts, sesame seeds, macadamia nuts, almonds, walnuts and brazil nuts. The specificity of the system was tested with DNA from more than 50 plant and animal species. The sensitivity of the method was suitable to detect allergenic ingredients in the low mg kg(-1) range. The limit of detection (LOD) for single allergens in different food matrices was 5 mg kg(-1). The novel analytical strategy represents a useful tool for the surveillance of established legislation on food allergens within the European Union. PMID- 19680916 TI - Monitoring of free glutamic acid in Malaysian processed foods, dishes and condiments. AB - A study to quantify the free glutamic acid content of six processed foods, 44 dishes and 26 condiments available in Malaysia was performed using high performance liquid chromatography with a fluorescence detector (HPLC-FRD). Recovery tests were carried out with spiked samples at levels from 6 to 31 mg g( 1). High recovery in different matrices was achieved ranging from 88% +/- 13% to 102% +/- 5.12%, with an average of 97% +/- 8.92%. Results from the study revealed that the average free glutamic acid content ranged from 0.34 +/- 0.20 to 4.63 +/- 0.41 mg g(-1) in processed foods, while in prepared dishes it was as low as 0.24 +/- 0.15 mg g(-1) in roti canai (puffed bread served with curry or dhal) to 8.16 +/- 1.99 mg g(-1) in dim sum (a small casing of dough, usually filled with minced meat, seafood, and vegetables, either steamed or fried). Relatively, the content of free glutamic acid was found to be higher in condiments at 0.28 +/- 0 mg g(-1) in mayonnaise to 170.90 +/- 6.40 mg g(-1) in chicken stock powder. PMID- 19680917 TI - Comparative study of three screening tests, two microbiological tube tests, and a multi-sulphonamide ELISA kit for the detection of antimicrobial and sulphonamide residues in eggs. AB - The screening of antimicrobial residues in eggs is an especially important subject. Three different commercial kits for the screening of sulphonamides and other antimicrobials in eggs were validated in accordance with Decision 2002/657/EC: one enzyme-linked immunoabsorbant assay (ELISA) kit multi sulphonamides (from RAISIO Diagnostics) and two microbiological tests (a Premi test from DSM and an Explorer kit from Zeu-Inmunotec). The false-positive rates were lower than 2% for all kits. The detection capabilities (CCbeta) have to be as low as possible for banned substances and lower than the maximum residue limit (MRL) when MRLs have been set. The sensitivity of the Premi test was better than that of the Explorer test, probably because of the dilution of the eggs before the Explorer test was used. The CCbeta values towards most of the tested sulphonamides were satisfactory with the Premi test (< or = 100 microg kg(-1)). Performance in a proficiency test for the detection of sulphonamides in eggs with the Premi test confirmed these results. The detection capabilities of tetracycline and doxycycline were at the level of the MRL or twice the MRL maximum. The detection capabilities for chlortetracycline and oxytetracycline were higher (four to six times the MRL). The detection capabilities for amoxicillin, neomycin, tylosin and erythromycin were lower than their respective MRLs. Detection capabilities for sulphonamides were much lower for the ELISA kit than for microbiological tests. The ELISA kit could be recommended for the targeted screening of sulphonamides in eggs. On the other hand, the Explorer and Premi tests could be used as wide screening tests allowing the detection of most of the antimicrobial families. PMID- 19680919 TI - Assessment of endogenous androgen levels in meat, liver and testis of Iranian native cross-breed male sheep and bull by gas chromatography-mass spectrometry. AB - Androgenic steroids always exist in different animal tissues at trace level, with significant numbers of interfering compounds, which makes their determination difficult. To solve some of the problems in quantification of the natural steroids in those tissues, a new GC-MS method was developed in this study. By using a surrogate analyte approach, which was developed in the authors' previous studies, and extensive sample preparation procedure, which successfully eliminates many of the interfering compounds and resulting in a cleaner extract, accuracy, precision, sensitivity and selectivity of the method for the determination of steroids in complex matrices such as meat, liver and testis were improved. By aid of this method, the levels of androgens in different tissues of Iranian native cross-breed bulls and male sheep were determined. According to the results obtained in the present study, although the androgenic profile (contents and ratios of precursors and metabolites to the main hormones) is similar between the same tissues of both animals, the total androgenic content of each tissue is higher in the bull than the same tissue in male sheep. In addition, in both animals higher amount of androgens were found in liver in comparison with meat and testis. PMID- 19680918 TI - Comparison of a fluoroquinolone surface plasmon resonance biosensor screening assay with established methods. AB - The performance of a previously developed immunochemical biosensor screening method for fluoroquinolone (FQ) antibiotics in poultry muscle, fish and egg was compared with established methods. Blank sample material of the target matrices was individually spiked with the FQs at half maximum residue levels. Homogeneity of the test materials was confirmed by liquid chromatography-mass spectrometry (LC-MS/MS). Identical sets of spiked samples as well as incurred samples from a previous feeding experiment were sent to three independent laboratories and analysed by LC-MS/MS, a microbiological assay and the new biosensor assay. The new method correctly identified all contaminated samples and demonstrated advantages in sensitivity and analysis time compared to the microbiological screening assay. PMID- 19680920 TI - Use of glyceroltriheptanoate as marker for processed animal by-products: development and validation of an analytical method. AB - A recently published European Regulation requires that the artificial marker, glycerol triheptanoate (GTH), be added to processed animal by-product (ABPs) prohibited from entering the food chain. The objective of this new requirement is to allow full traceability and ensure that these materials are disposed of in a proper way. Here, we report the development and single-laboratory validation of an analytical method for the determination of GTH in meat and bone meal plus animal fat. The method comprises three steps: (1) extraction of GTH from the samples with petroleum ether when analysing meat and bone meal or dissolving the sample in n-hexane when analysing fat; (2) clean-up of the extract using commercially available SPE cartridges; (3) determination of GTH by GC/MS or GC with flame ionisation detection (FID). The results of the validation study demonstrated that the relative standard for intermediate precision varied between 2.5 and 8.2%, depending on GTH concentration and the detector utilised. In all cases, the relative recovery rate was above 96%. The limit of quantification was 16 mg kg(-1) (GTH/fat content of the sample) with MS as detector and 20 mg kg(-1) with FID. Moreover, the method has been successfully applied in a second laboratory, indicating its transferability. Considering the minimum GTH concentration in ABPs of 250 mg kg(-1), the method is considered suitable for the intended purpose and can be utilised by EU Member States laboratories for official control and monitoring. PMID- 19680921 TI - Residues of rotenone, azadirachtin, pyrethrins and copper used to control Bactrocera oleae (Gmel.) in organic olives and oil. AB - Rotenone, azadirachtin, pyrethrins and copper fungicide decay curves were determined in olives and olive oil samples after experimental trials, consisting of one, two and three applications of each active ingredient, were carried out twice in 2005 and 2006. Rotenone, azadiracthin and pyrethrins were analyzed by extraction with acetonitrile and determined by liquid chromatography; copper was extracted into aqueous HCl and determined by chemical stripping. Pyrethrins were always found but in levels below the acceptable limits, whereas rotenone and copper residues always exceeded, after the pre-harvest interval, the maximum threshold allowed for olives. As regards residues in olive oil, rotenone was found in concentrations higher than those detected in olives. Copper accumulated in olives according to the number of applications, whereas, in oil, the residue was always lower than the maximum residue limit. The sensitivity of the method applied did not allow detection of azadirachtin. PMID- 19680923 TI - Impact of durum wheat milling on deoxynivalenol distribution in the outcoming fractions. AB - The milling behaviour of two naturally infected samples of durum wheat grain with contrasting levels of mycotoxins was studied. Although the two samples showed a similar milling behaviour, an increase of approximately 20% in deoxynivalenol (DON) levels was found in semolina from the sample containing the higher level of mycotoxin. However, even if the highest concentration of DON was found in fractions originating from the grain outer layers, the mycotoxin contamination in semolina and flours were not related to the amount of two compounds (ash or phytic acid) used to monitor these external tissues. The presence of the trichothecene-producing fungi in the inner-most semolina fraction was also shown using specific DNA primers and PCR amplification. Comparison of DON concentrations in the feed stock and corresponding output at each milling step or grinding of semolina fractions followed by sizing showed that concentration of mycotoxin occurs in the finest particles at the first processing steps. Therefore, DON contamination of milling fractions is not simply due to the presence of peripheral grain tissues. PMID- 19680922 TI - An isotope dilution headspace method with gas chromatography-mass spectrometry for determination of propylene oxide in food. AB - A method based on isotope dilution headspace and gas chromatography-mass spectrometry was developed for the determination of propylene oxide in foods. Optimum method sensitivity was achieved by the addition of NaCl in water at saturation and with the sample solution incubated at 90 degrees C. The method had good repeatability with relative standard deviations of 6.0, 7.6 and 2.2% at 5, 20 and 40 microg l(-1), respectively. The method was used to determine propylene oxide in 36 selected food composite samples from the 2007 Canadian total diet study. Propylene oxide was not detected in any samples analyzed with an average method detection limit of 0.5 ng g(-1). Hydrolysis of propylene oxide in water was observed as a first-order reaction with a half-life of 15 h at room temperature and less than 10 min at 90 degrees C. This confirms that it is very unlikely to find propylene oxide in foods as consumed due to its volatility and reaction with water. PMID- 19680924 TI - Fusarium mycotoxin content of UK organic and conventional wheat. AB - Each year (2001-2005), 300 samples of wheat from fields of known agronomy were analysed for ten trichothecenes by gas chromatography-mass spectrometry (GC/MS) including deoxynivalenol (DON), nivalenol, 3-acetyl-DON, 15-acetyl-DON, fusarenone X, T2 toxin, HT2 toxin, diacetoxyscirpenol, neosolaniol and T-2 triol and zearalenone by high-performance liquid chromatography (HPLC). Of the eleven mycotoxins analysed from 1624 harvest samples of wheat, only eight were detected, and of these only five-deoxynivalenol, 15-acetyl-DON, nivalenol, HT-2 and zearalenone-were detected above 100 microg kg(-1). DON was the most frequently detected Fusarium mycotoxin, present above the limit of quantification (10 microg kg(-1)) in 86% of samples, and was usually present at the highest concentration. The percentage of samples that would have exceeded the recently introduced legal limits varied between 0.4% and 11.3% over the five-year period. There was a good correlation between DON and zearalenone concentrations, although the relative concentration of DON and zearalenone fluctuated between years. Year and region had a significant effect on all mycotoxins analysed. There was no significant difference in the DON concentration of organic and conventional samples. There was also no significant difference in the concentration of zearalenone between organic and conventional samples, however organic samples did have a significantly lower concentration of HT2 and T2. Overall, the risk of UK wheat exceeding the newly introduced legal limits for Fusarium mycotoxins in cereals intended for human consumption is low, but the percentage of samples above these limits will fluctuate between years. PMID- 19680926 TI - Reduction of fumonisin B1 in extruded corn breakfast cereals with salt, malt and sugar in their formulation. AB - The objective was to determine the effect of added sodium chloride, barley malt and sucrose on the stability of fumonisin B1 (FB1) present in corn flour. Two levels of both sodium chloride (0.4% and 2%) and barley malt (0.8% and 5%) were added to the unextruded corn flour, and six levels of sucrose (3-10%) were used. The addition of sucrose at the lowest salt content (0.4%) as well as salt, either at 0.4% or at 2%, led to a significant decrease of FB1 levels in extruded samples, whereas malt, either at 0.8% or at 5%, did not significantly affect FB1 stability. Decontamination rates depended on the concentrations of added ingredients and ranged from 2% to 92%. The greatest reductions in FB1 content were achieved with extrusion cooking with a high salt content, whilst the lowest reductions were the result of processing corn flour with low contents of both salt and sucrose. Salt at 2% was the most effective ingredient in reducing FB1 content of the final extruded food. PMID- 19680925 TI - Occurrence of deoxynivalenol and its 3-beta-D-glucoside in wheat and maize. AB - Deoxynivalenol-3-beta-D-glucoside (D3G), a phase II plant metabolite of the mycotoxin deoxynivalenol (DON), occurs in naturally Fusarium-contaminated cereals. In order to investigate the frequency of occurrence as well as the relative and absolute concentrations of D3G in naturally infected cereals, 23 wheat samples originating from fields in Austria, Germany and Slovakia as well as 54 maize samples from Austrian fields were analysed for DON and D3G by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Both analytes were detected in all the 77 field samples. DON was found at levels from 42 to 4130 ng g(-1) (977 +/- 1000 ng g(-1) on average). The D3G concentrations in all cereal samples were in the range 10-1070 ng g(-1) (216 +/- 253 ng g(-1) on average), corresponding to about 5-46 mol% of their DON concentrations (15 +/- 8 mol% on average). PMID- 19680927 TI - Structural analysis of enniatin H, I, and MK1688 and beauvericin by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and their production by Fusarium oxysporum KFCC 11363P. AB - The molecular structures of enniatins H, I, and MK1688 and beauvericin were investigated by liquid chromatography-tandem mass spectrometry (LC-MS/MS). MS fragmentation occurred by loss of -CO after opening of the cyclic molecule to carbonyl carbon, and cleavage of the peptide and ester bonds in the molecular structure. Fusarium oxysporum KFCC 11363P was tested for its ability to produce beauvericin and enniatins H, I, and MK1688 on five cereal substrates: rice, barley, maize, wheat, and Indian millet kernels. Furthermore, optimal conditions for the production of the four mycotoxins by the Fusarium isolate were examined on maize at four temperatures (15, 20, 25, and 30 degrees C) and at three moisture contents (10, 20, and 40%). Large amounts of beauvericin and enniatin H were present in maize cultures at 25 degrees C (232.4 and 196.4 microg g(-1), respectively). Enniatins I and MK1688 were maximally formed at 20 degrees C (221.5 and 180.2 microg g(-1), respectively). The optimal moisture contents for the production of enniatins H (196.4 microg g(-1)) and MK1688 (165.6 microg g( 1)), were 40%. PMID- 19680928 TI - Moulds and mycotoxins in rice from the Swedish retail market. AB - A survey of moulds and mycotoxins was performed on 99 rice samples taken from the Swedish retail market. The main objective was to study the mould and mycotoxin content in basmati rice and rice with a high content of fibre. Samples of jasmine rice as well as long-grain rice were also included. The samples were analysed for their content of ochratoxin A (high-performance liquid chromatography (HPLC)), aflatoxin B(1), B(2), G(1), and G(2) (HPLC, RIDA(R)QUICK), and mould (traditional cultivation methods in combination with morphological analysis). The majority of samples were sampled according to European Commission Regulation 401/2006. Subsamples were pooled and mixed before milling and both mould and mycotoxin analyses were performed on milled rice. The results showed that the majority of basmati rice (71%) and many jasmine rice samples (20%) contained detectable levels of aflatoxin B(1) (level of quantification = 0.1 microg aflatoxin kg(-1) rice). Two samples of jasmine rice and ten basmati rice samples contained levels over the regulated European maximum limits of 2 microg kg(-1) for aflatoxin B(1) or 4 microg kg(-1) for total aflatoxins. Aspergillus was the most common mould genus isolated, but also Penicillium, Eurotium, Wallemia, Cladosporium, Epicoccum, Alternaria, and Trichotecium were found. The presence of Aspergillus flavus in 21% of the samples indicates that incorrect management of rice during production and storage implies a risk of mould growth and subsequent production of aflatoxin. Rough estimates showed that high rice consumers may have an intake of 2-3 ng aflatoxin kg(-1) bodyweight and day(-1) from rice alone. This survey shows that aflatoxin is a common contaminant in rice imported to Europe. PMID- 19680929 TI - New hierarchical classification of food items for the assessment of exposure to packaging migrants: use of hub codes for different food groups. AB - This paper describes development work undertaken to expand the capabilities of an existing two-dimensional probabilistic modelling approach for assessing dietary exposure to chemicals migrating out of food contact materials. A new three-level hub-coding system has been devised for coding different food groups with regards to their consumption by individuals. The hub codes can be used at three different levels representing a high, medium and low level of aggregation of individual food items. The hub codes were developed because they have a greater relevance to packaging migration than coding used (largely and historically) for nutritional purposes. Also, the hub codes will assist pan-europeanization of the exposure model in the future, when up to 27 or more different food coding systems from 27 European Union Member States will have to be assimilated into the modelling approach. The applicability of the model with the new coding system has been tested by incorporating newly released 2001 UK consumption data. The example used was exposure to a hypothetical migrant from coated metal packaging for foodstuffs. When working at the three hierarchical levels, it was found that the tiered approach gave conservative estimates at the cruder level of refinement and a more realistic assessment was obtained as the refinement progressed. The work overall revealed that changes in eating habits over time had a relatively small impact on estimates of exposure. More important impacts are changes over time in packaging usage, packaging composition and migration levels. For countries like the UK, which has sophisticated food consumption data, it is uncertainties in these other areas that need to be addressed by new data collection. PMID- 19680930 TI - Occurrence of 2,2,4-trimethyl-1,3-pentanediol monoisobutyrate (Texanol) in foods packed in polystyrene and polypropylene cups. AB - After simultaneous distillation-extraction (SDE) of foods packed in polystyrene (n = 77) and polypropylene cups (n = 42) from 61 different suppliers, coupled capillary gas chromatography-mass spectrometric (HRGC-MS) analyses indicated the presence of diastereomers of 2,2,4-trimethyl-1,3-pentanediol monoisobutyrate (TMPD-MIB; Texanol), a known coalescent of paints and printing inks. The contaminant was found in 55 and 50% of the polystyrene and polypropylene packed samples, respectively. Amounts ranged 1.2-64.5 microg kg(-1) in polystyrene cups (average 25.1 microg kg(-1)) and 0.9-45.7 microg kg(-1) in polypropylene cups (average 10.8 microg kg(-1)). The origin of Texanol in the printed plastic cups was demonstrated by separate HRGC-MS analysis, showing amounts in the higher microg kg(-1) range. In addition, the presence of two pairs of enantiomers, both found to be racemic by enantioselective multi-dimensional gas chromatography-mass spectrometry (enantio-MDGC-MS), excluded it being of natural origin. The detection limit of overall procedure (DLOP) and the reliable quantification limit (RQL) were 0.2 and 0.9 microg kg(-1), respectively. As the diester, 2,2,4 trimethyl-1,3-pentanediol diisobutyrate (TXIB), is on the EU list of regulated substances (restricted to single-use gloves only) with a migration limit of 5 mg kg(-1) in food and is metabolised rapidly by hydrolysis, the observed migration of the monoester Texanol at the microg kg(-1) level poses no risk of adverse effects. PMID- 19680931 TI - Cell transformation activities of abietic acid and dehydroabietic acid: safety assessment of possible contaminants in paper and paperboard for food contact use. AB - Abietic acid (AA) and dehydroabietic acid (DHA) have been detected in virgin paper products and recycled paper products used for food packaging. In order to evaluate the cell transformation activities of AA and DHA, the Bhas 42 cell transformation assay for initiation and promotion was carried out. Tested in the initiation stage, AA and DHA did not significantly increase transformation frequencies. On the other hand, both chemicals induced transformed foci dose dependently at the promotion stage. The highest transformed foci density induced by AA was about 13 foci/well at 60 nmol ml(-1), and that of DHA was about 16 foci/well at 40 nmol ml(-1) (solvent control = 2.3 +/- 1.4 foci/well). The present results suggest that AA and DHA may have tumour-promoting potential. PMID- 19680932 TI - Use of overall migration methodology to test for food-contact substances with specific migration limits. AB - This work investigated if overall migration test procedures could also be used to test for the migration of specific substances from plastics. The overall migration test procedure used was the evaporative gravimetric method used with volatile food simulants. Thirty food-contact substances (additives and monomers) were tested for their chemical stability and volatile loss during the heated evaporation stage of the overall migration procedure. Eighteen of the 30 were determined in an acceptable yield. It is concluded that in the list of approximately 620 European Union substances that have specific migration limits of 5 mg kg(-1) or higher, and based on considerations of stability and volatility, more than half could be amenable to control using overall migration methodology. This is particularly the case for inert plastics with low intrinsic overall migration values of oligomers. This means that based on the overall migration test result found, testing laboratories could decide on a case-by-case basis if known additives and starting substances are covered by the overall migration result and no separate testing would be required for specific migration, with time and resource cost savings. PMID- 19680933 TI - Formation of styrene during the Maillard reaction is negligible. AB - The elucidation of chemical pathways and the identification of intermediates leading to vinylogous compounds such as acrylamide by the Maillard reaction have proven challenging. This study was conducted to assess the formation of styrene from L-phenylalanine, employing binary mixtures of the amino acid heated together with simple C(3)-sugar analogue (1-hydroxyacetone) or methylglyoxal. The formation of the corresponding vinylogous product, i.e. styrene, was measured under different moisture, pH, and temperature conditions. The formation of intermediates over time was monitored by gas chromatography-mass spectrometry (GC MS) and liquid chromatography-mass spectrometry (LC-MS) together with the target compound styrene. Two intermediates, i.e. 1-phenethylaminopropan-2-one and 2 phenylethylamine, play a role in the formation of styrene, the latter of more importance in high-moisture systems, whilst the former favours the release of styrene in low-moisture systems. The model further showed that Strecker-type reactions are of less importance in the formation of styrene, as the yield from single immediate precursors was maximally 0.03 mol%. The low conversion rate of L phenylalanine to the vinylogous product and existing data on the occurrence of free L-phenylalanine in food plants suggests that the amounts of styrene expected in foods subjected to thermal treatment are negligible. PMID- 19680934 TI - Determination of synthetic drugs used to adulterate botanical dietary supplements using QTRAP LC-MS/MS. AB - Adulteration of botanical dietary supplements with prohibited synthetic drugs has become a serious problem. In this paper, a method for testing synthetic drugs used to adulterate botanical dietary supplements was developed using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) coupled with a linearity ion-trap system in the multiple reaction monitoring (MRM) plus enhanced product ion (EPI) mode. Twenty-three drugs exhibiting various pharmacological effects, comprising blood pressure and lipid-lowering agents, sedative drugs, anti-diabetic drugs, weight-reducing agents and aphrodisiac compounds, were studied. For all drugs, a single transition was monitored using protonated molecules as precursor ions. EPI spectra were stored in a library and recognized by library searching. Several undeclared drugs were identified in herbal remedies, e.g., glibenclamide, sibutramine hydrochloride and sildenafil. Overall, 35 positive samples were found out of a total of 105 botanical dietary supplements tested. The method was selective, sensitive, rapid, high-throughput and reliable. PMID- 19680935 TI - Levels of preservatives (sulfite, sorbate and benzoate) in New Zealand foods and estimated dietary exposure. AB - Thirty foods assessed as being the likely major contributors to dietary preservative exposure were purchased, prepared as normally consumed and analyzed for sulfite, sorbate and benzoate. The majority of preservative concentrations (>98%) were within maximum permitted levels (MPLs) specified in the Australia New Zealand Food Standards Code. Mean population level estimates of dietary exposure were well below the respective acceptable daily intakes (ADIs) for all age-gender groups for all preservatives at 7-27%, 1-4% and 1-8% of the ADI for sulfites, sorbates and benzoates, respectively. All population level 95th percentile estimates of dietary exposure were below the ADI, with the exception of estimates for sulfite exposure for 5-12-year-old boys. The results of the current survey indicate that dietary exposure to the preservatives, sulfite, sorbate and benzoate, represent a low level of public health risk. However, it should be noted that the exposure estimates determined in the current survey will be influenced by the assumptions made. PMID- 19680936 TI - Development and single-laboratory validation of an HPLC method for the determination of cyclamate sweetener in foodstuffs. AB - A method for the determination of cyclamate has been developed and single laboratory validated for a range of foodstuffs including carbonated and fruit juice drinks, fruit preserves, spreads, and dairy desserts. The method uses the peroxide oxidation of cyclamate to cyclohexylamine followed by derivatization with trinitrobenzenesulfonic acid and analysis by a modified reversed-phase high performance liquid chromatography-ultraviolet light (HPLC-UV). Cycloheptylamine is used as an internal standard. The limits of detection were in the range 1-20 mg kg(-1) and the analysis was linear up to 1300 mg kg(-1) cyclamic acid in foods and up to 67 mg l(-1) in beverages. Analytical recovery was between 82% and 123%, and results were recovery corrected. Precision was within experimentally predicted levels for all of the matrices tested and Horrat values for the combined standard uncertainty associated with the measurement of cyclamate between 0.4 (water-based drinks) and 1.7 (spreads). The method was used successfully to test three soft drink samples for homogeneity before analytical performance assessment. The method is recommended for use in monitoring compliance and for formal testing by collaborative trial. PMID- 19680937 TI - Core food of the French food supply: second Total Diet Study. AB - As first described in the 1980s, the core food intake model allows a precise assessment of dietary nutrient intake and dietary exposure to contaminants insofar as it reflects the eating habits of a target population and covers the most important foods in terms of consumption, selected nutrient and contaminant contribution. This model has been used to set up the sampling strategy of the second French Total Diet Study (TDS) with the aim of obtaining a realistic panorama of nutrient intakes and contaminant exposure for the whole population, useful for quantitative risk assessment. Data on consumption trends and eating habits from the second French individual food consumption survey (INCA2) as well as data from a 2004 purchase panel of French households (SECODIP) were used to identify the core foods to be sampled. A total of 116 core foods on a national scale and 70 core foods on a regional scale were selected according to (1) the consumption data for adults and children, (2) their consumer rates, and (3) their high contribution to exposure to one or more contaminants of interest. Foods were collected in eight French regions (36 cities) and prepared 'as consumed' to be analysed for their nutritional composition and contamination levels. A total of 20 280 different food products were purchased to make up the 1352 composite samples of core foods to be analysed for additives, environmental contaminants, pesticide residues, trace elements and minerals, mycotoxins and acrylamide. The establishment of such a sampling plan is essential for effective, high-quality monitoring of dietary exposure from a public health point of view. PMID- 19680938 TI - Presence and metabolism of endogenous androgenic-anabolic steroid hormones in meat-producing animals: a review. AB - The presence and metabolism of endogenous steroid hormones in meat-producing animals has been the subject of much research over the past 40 years. While significant data are available, no comprehensive review has yet been performed. Species considered in this review are bovine, porcine, ovine, equine, caprine and cervine, while steroid hormones include the androgenic-anabolic steroids testosterone, nandrolone and boldenone, as well as their precursors and metabolites. Information on endogenous steroid hormone concentrations is primarily useful in two ways: (1) in relation to pathological versus 'normal' physiology and (2) in relation to the detection of the illegal abuse of these hormones in residue surveillance programmes. Since the major focus of this review is on the detection of steroids abuse in animal production, the information gathered to date is used to guide future research. A major deficiency in much of the existing published literature is the lack of standardization and formal validation of experimental approach. Key articles are cited that highlight the huge variation in reported steroid concentrations that can result when samples are analysed by different laboratories under different conditions. These deficiencies are in most cases so fundamental that it is difficult to make reliable comparisons between data sets and hence it is currently impossible to recommend definitive detection strategies. Standardization of the experimental approach would need to involve common experimental protocols and collaboratively validated analytical methods. In particular, standardization would need to cover everything from the demographic of the animal population studied, the method of sample collection and storage (especially the need to sample live versus slaughter sampling since the two methods of surveillance have very different requirements, particularly temporally), sample preparation technique (including mode of extraction, hydrolysis and derivatization), the end-point analytical detection technique, validation protocols, and the statistical methods applied to the resulting data. Although efforts are already underway (at HFL and LABERCA) to produce more definitive data and promote communication among the scientific community on this issue, the convening of a formal European Union working party is recommended. PMID- 19680939 TI - Confirmatory method for the determination of various acetylgestagens in animal kidney fat using liquid chromatography-tandem mass spectrometry. AB - A confirmatory method has been developed and validated that allows for the simultaneous detection of medroxyprogesterone acetate (MPA), megestrol acetate (MGA), melengestrol acetate (MLA), chlormadinone acetate (CMA) and delmadinone acetate (DMA) in animal kidney fat using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The compounds were extracted from kidney fat using acetonitrile, defatted using a hexane wash and subsequent saponification. Extracts were then purified on Isolute CN solid-phase extraction cartridges and analysed by LC-MS/MS. The method was validated in animal kidney fat in accordance with the criteria defined in Commission Decision 2002/657/EC. The decision limit (CCalpha) was calculated to be 0.12, 0.48, 0.40, 0.63 and 0.54 microg kg(-1), respectively, for MPA, MGA, MLA, DMA and CMA, with respective detection capability (CCbeta) values of 0.20, 0.81, 0.68, 1.07 and 0.92 microg kg(-1). The measurement uncertainty of the method was estimated at 16, 16, 19, 27 and 26% for MPA, MGA, MLA, DMA and CMA, respectively. Fortifying kidney fat samples (n = 18) in three separate assays showed the accuracy of the method to be between 98 and 100%. The precision of the method, expressed as % RSD, for within-laboratory reproducibility at three levels of fortification (1, 1.5 and 2 microg kg(-1) for MPA, 5, 7.5 and 10 microg kg(-1) for MGA, MLA, DMA and CMA) was less than 5% for all analytes. PMID- 19680941 TI - Influence of physiological status on residues of lipophilic xenobiotics in livestock. AB - Data on the transfer of lipophilic xenobiotics from livestock feed and the environment to meat and milk are required for risk assessment purposes. Often, data are only available for lactating dairy cattle. Physiologically-based pharmacokinetic (PBPK) modeling has been used to explore differences between classes of food-producing animals. Blood and tissue levels of lipophilic xenobiotics under conditions approximating steady-state were simulated. Simulations of constant exposure were performed for lactating cows, non-lactating cattle, sheep, goats and swine. The tissue : blood partition coefficient, fat volume and fat blood flow were identified as critical determinants of predicted tissue concentrations. There may be significant breed differences in residues in milk and fat following exposure. Modeling was used to derive scaling factors that can be used to assist the extrapolation of transfer studies, carried out on lactating dairy cows, to other classes of cattle and different species. PMID- 19680940 TI - Occurrence of alpha- and beta-nortestosterone residues in the urine of injured male cattle. AB - The administration of anabolic steroids, for the purposes of growth promotion, to food-producing animals is banned in the EU. Among the compounds covered by this prohibition is ss-nortestosterone (beta-NT). This hormone is known to occur naturally in stallions and boars, and its main bovine metabolite, alpha nortestosterone (alpha-NT), occurs naturally in pregnant cows and neonatal calves. However, neither compound is believed to occur naturally in male cattle. During 2006, the presence of alpha-NT and, on occasion, beta-NT was confirmed in male cattle (bulls and steers) slaughtered in Northern Ireland on welfare grounds, as a result of acute injury. Subsequent investigations revealed no evidence of abuse at any of the farms involved and revealed that the phenomenon also occurred in three other regions of the EU, in similarly injured animals. A hypothetical link to release of the adrenal steroid, dehydroepiandrosterone (DHEA), in response to the stress of the injury was tested. Following the intravenous administration of DHEA to two normal steers, beta-NT (but not alpha NT) was confirmed in the urine of one steer. Thus, it may be concluded that both beta-NT and, by implication, alpha-NT can occur naturally in male cattle (or a specific cohort thereof) in contrast to previously accepted scientific knowledge. PMID- 19680942 TI - Application of enzyme-linked immunosorbent assay for quantification of the insecticides imidacloprid and thiamethoxam in honey samples. AB - An enzyme-linked immunosorbent assay (ELISA) was used for the determination of residues of imidacloprid and thiamethoxam insecticides in honey after simple dilution of the samples without either extraction or cleanup. The ELISA enabled accurate determination of imidacloprid and thiamethoxam down to limits of 20 and 5 ng g(-1) in honey, respectively. Average recoveries of imidacloprid and thiamethoxam from the fortified honey samples were 90-120 and 96-122%, and coefficients of variation ranged 5-12 and 3-15%, respectively. The results from the ELISA agreed well with those by liquid chromatography-mass spectrometry (LC MS) for the insecticide-spiked samples, with a correlation coefficient (r(2)) of 0.96 and a regression coefficient (slope) of 1.03. The results indicate that ELISA is a suitable tool for the quantification of imidacloprid and thiamethoxam in honey. PMID- 19680943 TI - Control strategies against grey mould (Botrytis cinerea Pers.: Fr) and corresponding fungicide residues in grapes and wines. AB - This study examines the most effective anti-Botrytis strategies leading to possible lower pesticides residues in wine. To provide wine growers with a number of high-quality solutions for protection against Botrytis for their vineyards while minimizing pesticide residues in the final product, various treatment approaches were tested. A total of 10 strategies with different specific fungicide treatments for controlling Botrytis cinerea were applied to grapes at different growing stages: flowering, bunch closure and colour change. The type of vine chosen was Gamay, as it is very sensitive to Botrytis cinerea. In each experimental plot, disease incidence and severity were assessed at harvest. In addition, pesticide residue analysis was carried out on grapes, musts and wines to monitor residue levels in each treatment and to follow changes at each stage of the wine-making process. A correlation was established between the efficiency of anti-Botrytis fungicide treatment and pesticide residues in wine. Several strategies using various fungicides showed good results in terms of treatment efficiency while minimizing pesticide residues in wine, thus providing interesting alternatives to limit the development of fungal resistance. PMID- 19680944 TI - Effect of particle size and spectral sub-range within the UV-VIS-NIR range using diffuse reflectance spectra on multivariate models in evaluating the severity of fusariosis in ground wheat. AB - Control (crops grown in natural conditions) and Fusarium head blight (FHB) damaged (crops inoculated with Fusarium culmorum conidia) grain of four wheat cultivars was ground and sieved into three fractions of different particle size. A series of blended samples differing in content of damaged material were prepared within fractions and cultivars, and diffuse reflectance spectra recorded within the 200-2500 nm wavelength range. Partial least-squares (PLS) models for the percentage of damaged material in blended samples were built for each of twelve series within different spectral ranges, and the root-mean-squared error of cross-validation (RMSECV) was used for the assessment of model performance. Errors using the models were lowest for the finest fraction independent of spectral range; however, their values depended on the cultivar. RMSECV for the finest fraction averaged over cultivars ranged from a little below 3.0 (when the ultraviolet light sub-range was used or participated with another one) to 8.1% (when only the near infrared (NIR) sub-range was used). For the medium and coarse fractions, averaged errors showed the same tendency of dependence on the sub range(s); however, with higher values that increased with an increase in particle size. In conclusion, within the different fractions of particle size and spectral ranges, the most sensitive to the presence of damaged material were models developed for the finest fraction and when the ultraviolet light sub-range was used in modelling. PMID- 19680946 TI - Fusarium mycotoxins in milling streams from the commercial milling of maize imported to the UK, and relevance to current legislation. AB - A study in three large commercial UK maize mills showed that Fusarium mycotoxins, such as deoxynivalenol, zearalenone and fumonisins present at mill intake, are distributed in milling streams approximately according to their occurrence in the maize seed structure. Fractions derived from the endosperm tended to contain the lowest levels of mycotoxins. Concentrations of mycotoxins within the endosperm are also related to the particle size. However, the products derived from the embryo or outer seed layers contained the highest mycotoxin levels being concentrated up to five times or more, although these components are normally used for animal feed or industrial use. The general pattern of mycotoxin distribution found when milling French and Argentinean maize was similar, although very variable, and it is concluded that this variability stems from different milling strategies used at each mill and from the nature and condition of each consignment of maize. Mycotoxins in maize grits (particle sizes >500 microm) were usually reduced by the greatest amount when compared with the whole maize, while flour (< or =500 microm) could be both reduced or increased depending on the mill and consignment. Thus, in most situations mycotoxin concentrations in whole maize that meet European Commission legislation on intake should give rise to levels in milled ingredients that should also do so. However, this was not always true in some ingredients, especially for fumonisins in those fractions with particle size < or =500 microm. PMID- 19680945 TI - In vitro and in vivo characterization of mycotoxin-binding additives used for animal feeds in Mexico. AB - The study was conducted to characterize and compare twelve different additives distributed in Mexico as mycotoxin binders utilizing: (1) equilibrium isothermal analysis for aflatoxin B(1) (AFB(1)) adsorption, (2) a variety of mineralogical probes, and (3) Hydra toxicity bioassay. The test additives Milbond-TX (MLB), Mycoad (MCA), Volclay FD181 (VOL), Fixat (FXT), Toxinor (TOX), Mexsil (MEX), Mycosil (MYC), Klinsil (KLS), Zeotek (ZEO), Duotek (DUO), Mycosorb (MSB), and Mycofix Plus 3.0 (MIX) were compared with NovaSil Plus (NSP). Isotherms for AFB(1) adsorption were conducted at pH 2 and pH 6.5, mimicking pH conditions in the stomach and small intestine. Mineralogical analysis included determination of swelling volume, X-ray diffraction analysis, and fractionation procedures. A Hydra vulgaris toxicity study was performed to evaluate the potential safety of the additives. Computer-generated isotherm data were fit using the Langmuir model, and parameters of Q(max) and K(d) were estimated. The most effective additives for AFB(1) at both pH conditions were NSP, MLB, MCA and VOL, while the least effective was MSB. The amounts of sand, silt and clay fractions varied among the additives. Nine of the additives showed the presence of smectite. Most of the additives were found to be non-toxic to Hydra except for the organoclays (ZEO, DUO) and MSB. In general, NSP demonstrated the highest sorption capacity in the bulk material and the different fractions. Studies to characterize these binding additives further and to evaluate their multiple mycotoxin sorption claims are ongoing. PMID- 19680948 TI - Off-flavours in wines through indirect transfer of volatile organic compounds (VOCs) from coatings. AB - This paper assesses the impact of volatile organic compounds (VOCs) from the drying of coatings on the sensory characters of corks and wines. According to Italian National Standard Method 11021:2002, a small-scale chamber was used (1) to expose wines to the drying of coatings with both low and high VOCs, and (2) to expose corks to the same coatings. After exposure to the coatings, the corks were then placed in direct contact with wine. Different styles of white, red and rose wines were tested. In both directly exposed wines and in wines after contact with the exposed cork, the taste and smell off-flavour perception and intensity were assessed by a panel of eight experienced wine tasters using a five-point numerical scale according to International Organization for Standardization (ISO) standard methods. The results showed that the sensory characters of wines, especially taste, were influenced by the VOC content of the coatings. The taste off-flavour perception was found to be higher than the smell in wines exposed to coatings with either high or low VOCs contents. Analysis of variance (ANOVA) and Duncan analysis prove that: (1) panellist's answers were significantly different, (2) it was difficult to differentiate the off-flavour perception on the high level scale, and (3) the panellist off-flavour perceptions versus wine style discriminated the wines into two groups (red and white/rose). For all the wine styles, Pearson's test showed no significant correlation between off-flavour perception levels and the main chemical characters of the wines. For the wines that were in direct contact with the exposed corks, the panellists detect the off flavours according to the levels of VOCs in the coating and wine styles and they assessed the highest levels of alteration were to the taste. PMID- 19680947 TI - Quantitative analysis of patulin in apple juice by thin-layer chromatography using a charge coupled device detector. AB - A method was developed and validated in-house for the detection and quantification of patulin in apple juice concentrate using a charge coupled device (CCD) on thin-layer chromatography (TLC) plates. Samples were extracted with ethyl acetate and then cleaned-up by extraction with a sodium carbonate solution. The method showed a mean recovery of 95%. The quantification and detection limit were 14 microg l(-1) and 0.005 microg per spot, respectively. The CCD camera is sufficiently sensitive to detect changes in spot fluorescence intensity caused by small differences in mycotoxin concentration under homogeneous illumination from a UV light source. The results of validation confirmed the efficiency of the method, which is sensitive enough to be used to quantify patulin in apple juice by producers or for government monitoring/survey programs. The method was applied to the analysis of 16 apple juice concentrate samples and patulin levels ranged from 15 to 46 microg l(-1). This study demonstrated the applicability of the TLC-CCD technique as a tool for monitoring patulin in apple juice. PMID- 19680949 TI - Effect of cooking or handling conditions on the furan levels of processed foods. AB - The aim of this study was to investigate the possible effects of cooking or handling conditions on the concentration of furan in processed foods. The analytical method used to analyse furan levels in foods was optimized based on solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS). In baby soups, the concentration of furan decreased by up to 22% after opening a lid for 10 min. In the baby food in retort packaging, the level of furan was reduced by 15-33% after heating the foods at 50 degrees C without a lid. Furan in rice seasonings was evaporated completely after heating the foods at 60 degrees C. Regarding powered milk, the levels of furan were too low to be compared under various conditions. The levels of furan decreased to 58% in beverage products for babies, after storing them at 4 degrees C for 1 day without a lid. The levels of furan in canned foods such as cereal and vegetable were reduced by zero to 52% when they were stored without stirring in a refrigerator at 4 degrees C for 1 day. When we boiled canned fish, the furan present was almost completely evaporated. It is recommended that canned meats be heated up to 50-70 degrees C for the reduction (26-46%) of furan levels. The levels of furan in instant and brewed coffee samples were significantly reduced after storing for 11 to 20 min at room temperature without a lid (p < 0.05). PMID- 19680950 TI - Risk assessment of furan in commercially jarred baby foods, including insights into its occurrence and formation in freshly home-cooked foods for infants and young children. AB - Furan is a possible human carcinogen (IARC group 2B) with widespread occurrence in many types of foods. In this study, a survey of furan contamination in 230 commercially jarred ready-to-eat infant food products was conducted using headspace sampling in combination with gas chromatography and mass spectrometry (HS-GC/MS) with a detection limit of 0.2 microg kg(-1). The incidence of furan contamination in jarred infant beverages, cereals and fruits was relatively low, with average concentrations below 10 microg kg(-1). Significantly higher concentrations were found in pasta (34.8 +/- 14.5 microg kg(-1)), meals containing meat (28.2 +/- 15.0 microg kg(-1)), and meals containing vegetables (31.2 +/- 17.3 microg kg(-1)). The average exposure of 6-month-old infants to furan was estimated to be 0.2 microg per kg bodyweight per day. The margin of exposure calculated using the T25 dose descriptor would be 2692, which points to a possible public health risk. In contrast to commercially jarred food products, none of 20 freshly home-prepared baby foods contained furan above the limit of detection. Only after re-heating in closed vessels was furan found to have formed. Furan was especially prevalent in reheated foods containing potatoes, with values ranging between 2.3 and 29.2 microg kg(-1). The formation of furan in potato-containing baby foods was increased by addition of ascorbic acid, by longer heating times above 1 h and by temperatures above 50 degrees C. Research regarding reduction of furan in commercial baby foods should be conducted, with a priority aimed at reducing this heat-induced contaminant without concomitantly increasing the microbiological risk. PMID- 19680951 TI - Analysis of furan in coffee of different provenance by head-space solid phase microextraction gas chromatography-mass spectrometry: effect of brewing procedures. AB - A simple, sensitive and accurate method for the analysis of furan in roasted coffee has been used based on headspace-solid-phase micro-extraction (HS-SPME) coupled to gas chromatography-mass spectrometry (GC-MS). The extraction was performed using 75-microm carboxen/polydimethylsiloxane fiber. Ionic strength, extraction time and temperature, and desorption time were assessed as the most important parameters affecting the HS-SPME procedure and d(4)-furan was used as the internal standard. The linearity range was in the range 0.0075-0.486 ng g( 1); the LOD and LOQ calculated using the signal-to-noise ratio approach were 0.002 and 0.006 ng g(-1), respectively. The inter- and intra-day precision was 8 and 10%, respectively. The concentration of furan found in batches of roasted coffee powder different producing countries ranged from 57.3 to 587.3 ng g(-1). The mean reduction in furan levels observed when brewing coffee by either infusion, using a moka pot or an expresso machine was 57, 67.5 and 63.3%, respectively. PMID- 19680952 TI - Effects of various cooking conditions on acrylamide formation in rolled patty. AB - In this study on acrylamide formation, the effects of the type of frying oil, frying period and covering with egg during frying of a rolled patty (a traditional Turkish carbohydrate-rich food) were investigated. The differences between frying periods were statistically significant for each oil (p < 0.01). For comparable frying periods, the maximum acrylamide content was found in the rolled patties fried with sunflower oil, and the minimum acrylamide content was found in the rolled patties fried with corn oil. A decrease of 39-65% in acrylamide formation in the rolled patties covered with egg was found for each of the three types of oil. In addition, a high linear correlation (R > 0.90) was found between L (light) values and acrylamide amounts. PMID- 19680954 TI - Aniline in vegetable and fruit samples from the Canadian total diet study. AB - An isotope dilution method based on solvent extraction followed by GC-MS analysis was developed and used to determine aniline in vegetable and fruit samples collected from the Canadian total diet study. Aniline was not detected in any of the 23 vegetable samples from the 2005 total diet study at a method detection limit of 0.01 mg kg(-1). Among the 16 fruit samples, it was detected only in apple samples, with an average concentration of 0.278 mg kg(-1). Aniline was not detected in apple samples collected in the 2002, 2003, 2006 or 2007 total diet studies, but it was detected in the apple samples collected from the 2001 and 2004 studies, at concentrations of 0.085 and 0.468 mg kg(-1), respectively. The average aniline concentration for the 2001, 2004 and 2005 apple samples was 0.277 mg kg(-1). Good repeatability of the method was observed with replicate analysis of apple samples, with relative standard deviations (RSD) ranging 3.8-21% and an average of 11%. PMID- 19680955 TI - Evaluation of the illegal use of clenbuterol in Portuguese cattle farms from drinking water, urine, hair and feed samples. AB - The recent discovery of clenbuterol contamination in Portuguese food led to the specific inspection of 16 cattle farms for beta-agonists, involving the analysis of a total of 486 samples (78 feed, 106 drinking water, 168 urine and 134 hair). The samples were screened for the beta-agonists: bromobuterol, cimaterol, clenbuterol, clenpenterol, clenproperol, hydroxymethylclenbuterol, mapenterol, salbutamol and terbutaline. Only clenbuterol was found in all analyzed matrices and the most likely method of illegal administration to animals was through drinking water. Of all samples analysed, 14.15% of drinking water were found positive in the range 0.03-3.80 mg l(-1) clenbuterol. Inclusion of hair samples in the Portuguese plan for clenbuterol residue control in live animals is discussed. PMID- 19680953 TI - Development of enzyme-linked immunosorbent assays for Sudan dyes in chilli powder, ketchup and egg yolk. AB - This study aimed at developing sensitive competitive enzyme-linked immunosorbent assays (ELISAs) for the banned Sudan dyes using polyclonal antibodies. Three different formats were developed and characterized in terms of sensitivity, selectivity and rapidity. A competitive indirect ELISA was developed, which showed an IC(50) of 3.8 microg l(-1). Two competitive direct ELISAs were also developed, in which the antibody was added before or simultaneously with the other reagents; the first showed an IC(50) of 8.3 microg l(-1) and the latter showed an IC(50) of 4.9 microg l(-1). Nevertheless, considering dilution of extracts which is needed to offset matrix interference, the limits of detection of the three formats were substantially the same (10 microg kg(-1)). The antibodies in all three test formats were able to recognize Sudan I and partially Sudan II, III and IV; no cross-reactivity was observed with the five edible dyes. Twenty food samples, including chilli powder, paprika, ketchup, and egg, were extracted by a simple sample preparation and very limited dilution. Extracts were analyzed by the developed competitive direct ELISA with the simultaneous addition of reagents. A good correlation was observed (y = 1.19 x-10.0, r(2) = 0.991, n = 20) when the data was compared with that obtained through a conventional HPLC method. PMID- 19680956 TI - Dietary exposure to lead by children and adults in the Jinhu area of China. AB - This paper reports an estimate of the dietary intake of lead by children and adults from the Jinhu area in South-eastern China. A duplicate diet approach was used to collect food samples in November and December 2007. The lead content was measured by furnace atomic absorption spectrometry. Median lead intakes for different groups were as follows: children: 1.65 microg(kg(bw))(-1) day(-1); and adults: 1.10 microg(kg(bw))(-1) day(-1). No exposure exceeded the provisional tolerable weekly intake (PTWI) of 25 microg(kg(bw))(-1) week(-1) proposed by the World Health Organization (WHO). However, median and maximum exposure for the children group amounted to 60.8% and 97.2% of the PTWI, respectively, which were higher than those for adults, which were 40.5% and 86.4%, respectively. It was concluded that health risks due to dietary lead intake seem to be high in Jinhu. PMID- 19680957 TI - Optimizing conditions for methylmercury extraction from fish samples for GC analysis using response surface methodology. AB - Response surface methodology (RSM) was used to determine the optimum experimental conditions to extract methylmercury from fish samples for GC analysis. The influence of four variables - acid concentration (3-12 M), cysteine concentration (0.5-2% w/v), solvent volume (3-9 ml) and extraction time (10-30 min) - on recovery of methylmercury was evaluated. The detection limit for methylmercury analysis using a microelectron capture detector was 7 ng g(-1) in fish samples. The mean recovery under optimum conditions was 94%. Experimental data were adequately fitted into a second-order polynomial model with multiple regression coefficients (r(2)) of 0.977. The four variables had a significant effect (p < 0.05) on the recovery of methylmercury from a reference material (BCR-463). Optimum conditions for methylmercury extraction were found using an acid concentration of 12.2 M, cysteine concentration of 2.4%, solvent volume of 1.5 ml and extraction time of 35 min. The validation of the developed method to analyze methylmercury in fish samples exhibited good agreement with mercury content in the samples. PMID- 19680958 TI - Determination of arsenic in foods by flow injection on-line sorption pre concentration with hydride generation atomic fluorescence spectrometry. AB - A method was developed for the determination of total arsenic in foods using flow injection on-line sorption coupled with hydride generation atomic fluorescence spectrometry (HG-AFS) using a cigarette filter as the sorbent material. After reducing As(V) to As(III) by using L-cysteine, the determination of total arsenic was achieved through on-line formation and retention of the pyrrolidine dithiocarbamate arsenic complex (As(III)-PDC) on the cigarette filter, which was packed in the pre-concentration column and total arsenic was determined by HG AFS. The analytes were eluted with 1.68 mol l(-1) HCl from the sorbent material. With consumption of 22 ml of the sample solution, an enrichment factor of 25.6 was obtained at a sample throughput of 11.6 h(-1). The detection limits (3 standard deviations) and the precision (relative standard deviation) in foods ranged from 2.5 to 9.9 ng g(-1) and from 1.1 to 2.2%, respectively. The method was used to determine arsenic in carrot, mushroom, chicken tissue, cod fish, rice, common carp and shrimp. PMID- 19680959 TI - Correlation of lead, cadmium and mercury levels in tissue and liver samples with age in cattle. AB - The aim of this study was to determine the accumulation of selected heavy metals (Pb, Cd, Hg, As) in meat and liver of cattle. The animals were divided into four age-groups which allowed the analysis of statistical-mathematical correlations between the age of the animals and contamination of meat. The research material for determination of heavy metal levels was taken from the longissimus back muscle (m. longissimus dorsi) and samples from the tail lobe of the liver. Analysis showed that contamination by Cd and Pb is clearly dependent on the age of the animal. PMID- 19680960 TI - Pesticide residues in apples grown under a conventional and integrated pest management system. AB - This paper describes method validation for determination of more than 40 pesticides in apples using a GC technique. Target compounds belonged to the organochlorine, organophosphorus, carbamates, pyrethroids, triazoles, dicarboximides and strobilurins groups, among others. Sample preparation consisted of acetone extraction and subsequent cleanup/concentration by SPE with a polymer-based sorbent. Single quadrupole GC-MS operating in SIM mode and electron impact ionization was used for identification and quantification of the pesticides. Average recoveries for analytes ranged between 70 and 110% at three fortification levels - 0.01, 0.1 and 0.2 mg kg(-1). Relative standard deviations were lower than 20% for all tested compounds. Calculated limits of detection and limits of quantification were below 0.01 mg kg(-1), which were sufficiently low compared to the maximum residue levels (MRLs) established by European legislation. The proposed method was applied for determination of pesticide residue in four selected apple varieties after harvesting. Whole and processed fruits (peel and peeled fruits) were analyzed from different treatment systems: two conventional, one based on integrated pest management (IPM) and two variants based on organic production (controls). Higher levels of pesticide residues were found in apple fruits under conventional conditions. Fenitrothion and chlorpyrifos residues were detected frequently in apple peel at concentrations up to 0.45 and 0.77 mg kg(-1), respectively. The levels found in the whole fruits of the same samples were much lower than in peel and below the respective MRLs (0.5 mg kg(-1) for both pesticides). Measurable residues of triadimenol only, up to 0.05 mg kg(-1), concentrated in the peel, were found in the apples from IPM. PMID- 19680961 TI - Multiresidue method for the determination of 77 pesticides in wine using QuEChERS sample preparation and gas chromatography with mass spectrometry. AB - A method based on a QuEChERS (quick, easy, cheap, effective, rugged, safe) sample preparation method and gas chromatography with mass spectrometric detection by selected ion monitoring (GC/MS-SIM) was developed for simultaneous determination of 77 pesticide residues in wine. An extraction of 10 ml of sample with acetonitrile followed by liquid-liquid partition formed by the addition of 4 g MgSO(4) and 3 g NaCl was applied in the sample preparation. The clean-up was carried out by applying dispersive solid-phase with 150 mg MgSO(4) as well as 50 mg primary secondary amine (PSA). One quantitation ion and at least two identification ions were selected in the analytical method for each pesticide compound by GC/MS. The recovery data were obtained by spiking blank samples at two concentration levels (0.05 and 0.2 mg l(-1)). The recoveries of all pesticides were in the range 70-110%, with intra-day precision of less than 15%, and the inter-day precision of less than 22% and 15% for 0.05 and 0.2 mg l(-1) fortification levels, respectively. Linearity was between 0.02 and 2 mg l(-1) with determination coefficients (R(2)) greater than 0.98 for all compounds. The limits of quantification (LOQs) for the 77 pesticides ranged from 0.003 to 0.05 mg l(-1). This method was applied for routine analysis in market products. PMID- 19680962 TI - Geostatistical analysis of the spatial distribution of mycotoxin concentration in bulk cereals. AB - Deoxynivalenol (DON) and ochratoxin A (OTA) in agricultural commodities present hazards to human and animal health. Bulk lots are routinely sampled for their presence, but it is widely acknowledged that designing sampling plans is particularly problematical because of the heterogeneous distribution of the mycotoxins. Previous studies have not explicitly looked at the interactions between the spatial distribution of the mycotoxin and the strategy used to take samples from bulk. Sampling plans are therefore designed on the assumption of random distributions. The objective of this study was to analyse the spatial distribution of DON and OTA in bulk commodities with geostatistics. This study was the first application of geostatistical analysis to data on mycotoxins contamination of bulk commodities. Data sets for DON and OTA in bulk storage were collected from the literature and personal communications, of which only one contained data suitable for geostatistical analysis. This data set represented a 26-tonne truck of wheat with a total of 100 sampled points. The mean concentrations of DON and OTA were 1342 and 0.59 microg kg(-1), respectively. The results showed that DON presented spatial structure, whilst OTA was randomly distributed in space. This difference between DON and OTA probably reflected the fact that DON is produced in the field, whereas OTA is produced in storage. The presence of spatial structure for DON implies that sampling plans need to consider the location of sample points in addition to the number of points sampled in order to obtain reliable estimates of quantities such as the mean contamination. PMID- 19680963 TI - Molecularly imprinted solid-phase extraction of fumonisin B analogues in bell pepper, rice and corn flakes. AB - A molecularly imprinted polymer (MIP) for the recognition of fumonisin B analogues (FB) using 2-(diethylamino) ethyl methacrylate (DEAEM) as functional monomer and trimethylolpropane trimethacrylate (TRIM) as cross-linker was prepared by bulk polymerization in acetonitrile. Fumonisin B(1) (FB(1)) was used as a template molecule. A molecularly imprinted solid-phase extraction (MISPE) procedure was developed for further application in the analysis of FB. The performance of the MIP throughout the clean-up of spiked bell pepper, rice and corn flake sample extracts was compared with the results obtained when using non imprinted polymer, C(18), strong anion exchange and immunoaffinity sorbents. Extracts were analysed for FB with liquid chromatography-tandem mass spectrometry (LC-MS/MS) after clean-up. Depending on the food matrix and the concentration range of the fumonisins, recoveries after MISPE varied from 62 to 86%, from 62 to 83%, and from 67 to 81% for fumonisin B(1) (FB(1)), fumonisin B(2) (FB(2)) and fumonisin B(3) (FB(3)), respectively. The selectivity of the synthesized MIP for mycotoxins belonging to the group of FB was confirmed by evaluating cross reactivity from analogue structures and other mycotoxins. Analysis of 39 naturally contaminated samples (corn flakes) by liquid chromatography tandem mass spectrometry indicated that the synthesized MIP could be an excellent alternative for clean-up and pre-concentration of FB in food samples. Pearson correlations between immunoaffinity clean-up and MISPE were calculated and amounted to 0.923 for FB(1), 0.808 for FB(2), and 0.759 for FB(3). It was shown that the developed MIP could be reused more than 50 times. The synthesis of an FB(1) imprinted polymer and its application in food analysis is reported for the first time. PMID- 19680964 TI - LC-MS/MS multi-analyte method for mycotoxin determination in food supplements. AB - A multi-analyte method for the liquid chromatography-tandem mass spectrometric determination of mycotoxins in food supplements is presented. The analytes included A and B trichothecenes (nivalenol, deoxynivalenol, 3 acetyldeoxynivalenol, 15-acetyldeoxynivalenol, neosolaniol, fusarenon-X, diacetoxyscirpenol, HT-2 toxin and T-2 toxin), aflatoxins (aflatoxin-B(1), aflatoxin-B(2), aflatoxin-G(1) and aflatoxin-G(2)), Alternaria toxins (alternariol, alternariol methyl ether and altenuene), fumonisins (fumonisin B(1), fumonisin-B(2) and fumonisin-B(3)), ochratoxin A, zearalenone, beauvericin and sterigmatocystin. Optimization of the simultaneous extraction of these toxins and the sample pretreatment procedure, as well as method validation were performed on maca (Lepidium meyenii) food supplements. The results indicated that the solvent mixture ethyl acetate/formic acid (95:5, v/v) was the best compromise for the extraction of the analytes from food supplements. Liquid-liquid partition with n-hexane was applied as partial clean-up step to remove excess of co extracted non-polar components. Further clean-up was performed on Oasis HLB cartridges. Samples were analysed using an Acquity UPLC system coupled to a Micromass Quattro Micro triple quadrupole mass spectrometer equipped with an electrospray interface operated in the positive-ion mode. Limits of detection and quantification were in the range of 0.3-30 ng g(-1) and 1-100 ng g(-1), respectively. Recovery yields were above 60% for most of the analytes, except for nivalenol, sterigmatocystine and the fumonisins. The method showed good precision and trueness. Analysis of different food supplements such as soy (Glycine max) isoflavones, St John's wort (Hypericum perforatum), garlic (Allium sativum), Ginkgo biloba, and black radish (Raphanus niger) demonstrated the general applicability of the method. Due to different matrix effects observed in different food supplement samples, the standard addition approach was applied to perform correct quantitative analysis. In 56 out of 62 samples analysed, none of the 23 mycotoxins investigated was detected. Positive samples contained at least one of the toxins fumonisin-B(1), fumonisin-B(2), fumonisin-B(3) and ochratoxin A. PMID- 19680965 TI - Changes in ochratoxin A and type B trichothecenes contained in wheat flour during dough fermentation and bread-baking. AB - Ochratoxin A (OTA) and type B trichothecenes are mycotoxins that occur frequently in cereals and thus can be found in cereal by-products such as bread. The aim of this work was to study the variation of the levels of OTA, deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON) and nivalenol (NIV) during the bread-making process. This was done by using wheat flour spiked with different levels of toxins. Mycotoxin levels were controlled after fermentation of the dough with yeasts (Saccharomyces cerevisiae) and after further baking at different temperature-time combinations. Analysis of variance (ANOVA) of the results showed a significant reduction in OTA level (p < 0.05) during fermentation of the dough. The reduction ranged between 29.8% and 33.5%, depending on the initial concentration of toxin in the flour. During this period, the level of the other mycotoxins studied was not modified. By contrast, in the baking phase there were significant changes in the levels of the four mycotoxins, although the reduction was similar under all the baking conditions. Considering all the temperature-time conditions tested, it can be concluded that during the baking period the average reduction of OTA, NIV, 3-ADON, and DON was 32.9%, 76.9%, 65.6%, and 47.9%, respectively. PMID- 19680966 TI - Rapid and non-invasive analysis of deoxynivalenol in durum and common wheat by Fourier-Transform Near Infrared (FT-NIR) spectroscopy. AB - Fourier transform near-infrared spectroscopy (FT-NIR) was used for rapid and non invasive analysis of deoxynivalenol (DON) in durum and common wheat. The relevance of using ground wheat samples with a homogeneous particle size distribution to minimize measurement variations and avoid DON segregation among particles of different sizes was established. Calibration models for durum wheat, common wheat and durum + common wheat samples, with particle size <500 microm, were obtained by using partial least squares (PLS) regression with an external validation technique. Values of root mean square error of prediction (RMSEP, 306 379 microg kg(-1)) were comparable and not too far from values of root mean square error of cross-validation (RMSECV, 470-555 microg kg(-1)). Coefficients of determination (r(2)) indicated an "approximate to good" level of prediction of the DON content by FT-NIR spectroscopy in the PLS calibration models (r(2) = 0.71 0.83), and a "good" discrimination between low and high DON contents in the PLS validation models (r(2) = 0.58-0.63). A "limited to good" practical utility of the models was ascertained by range error ratio (RER) values higher than 6. A qualitative model, based on 197 calibration samples, was developed to discriminate between blank and naturally contaminated wheat samples by setting a cut-off at 300 microg kg(-1) DON to separate the two classes. The model correctly classified 69% of the 65 validation samples with most misclassified samples (16 of 20) showing DON contamination levels quite close to the cut-off level. These findings suggest that FT-NIR analysis is suitable for the determination of DON in unprocessed wheat at levels far below the maximum permitted limits set by the European Commission. PMID- 19680967 TI - Brominated dioxins (PBDD/Fs) and PBDEs in marine shellfish in the UK. AB - The occurrence of brominated dioxins (PBDD/Fs) and polybrominated diphenyl ethers (PBDEs) was investigated in commonly consumed species of marine shellfish in the UK. Individual samples of Pacific oysters (Crassostrea gigas), native oysters (Ostrea edulis), mussels (Mytilus edulis), scallops (Pecten maximus), and cockles (Cerastoderma edule) were collected from different coastal regions between 2006 and 2007. Samples of a particular species from each site were composited and 60 samples were analysed. Polybrominated dibenzofurans (PBDFs) occurred more frequently and generally at a higher level than polybrominated dibenzodioxins (PBDDs), except for 237-TriBDD, which was the predominant PBDD/F congener in some species, notably oysters. This profile may reflect the environmental distribution of these compounds and the effects of removal mechanisms, such as degradation, selective uptake and metabolism. PBDEs were detected in all samples. The dominant congeners were BDEs 47, 49, 99 and 100 and, to a lesser extent, BDEs 66 and 154. The occurrence of BDE-209 was observed in most samples and appears to be species selective, with the highest values occurring almost exclusively in mussels and cockles. Among the species studied, oysters and mussels displayed relatively higher levels of both sets of contaminants; native oysters, in particular, showed elevated levels of 237-TriBDD (up to 14.5 ng/kg). In general, contaminant levels appeared to be consistent with the extent of local industrialisation with lower levels observed in more remote areas such as the north of Scotland. Polybrominated biphenyls (PBBs) were also measured, and PBBs 49, 52 and 77 were the most frequently detected, although levels were very low. Dietary intakes, estimated for PBDD/Fs, showed that 237-TriBDD from single portions of oysters constituted a high proportion of the total dietary intake of the congener but, otherwise, dietary intakes of PBDD/Fs from shellfish were relatively low. PMID- 19680969 TI - Migration study of carvacrol as a natural antioxidant in high-density polyethylene for active packaging. AB - The migration behaviour of low molecular weight compounds from food packaging materials is one of the key issues in assessing the possibility of use in such applications. The aim of this work was to study the migration of carvacrol (1% and 2% w/w) when added to high-density polyethylene. All materials were exposed to the food simulants olive oil and distilled water separately at 40 degrees C and 25 degrees C. Three significant variables influencing the migration process were considered: incubation temperatures, the initial concentration of antioxidant, and the type of simulant (oil and aqueous). The amount of carvacrol migrating to olive oil was significantly higher than in water because of the higher solubility of this antioxidant in oil. Experimental results agreed reasonably well with those obtained by the application of a simple model derived from Fick's Second Law. Carvacrol could therefore be used in active packaging formulations as its release from the polymer matrix can be controlled. PMID- 19680968 TI - Migration of bisphenol A from plastic baby bottles, baby bottle liners and reusable polycarbonate drinking bottles. AB - Human exposure to bisphenol A (BPA) has recently received special attention. It has been shown that exposure to BPA may occur through the consumption of beverages or foods that have been in contact with polycarbonate (PC) plastic containers or epoxy resins in food packaging. A BPA migration study was conducted using a variety of plastic containers, including polycarbonate baby bottles, non PC baby bottles, baby bottle liners, and reusable PC drinking bottles. Water was used to simulate migration into aqueous and acidic foods; 10% ethanol solution to simulate migration to low- and high-alcoholic foods; and 50% ethanol solution to simulate migration to fatty foods. By combining solid-phase extraction, BPA derivatization and analysis by GC-EI/MS/MS, a very low detection limit at the ng l(-1) level was obtained. Migration of BPA at 40 degrees C ranged from 0.11 microg l(-1) in water incubated for 8 h to 2.39 microg l(-1) in 50% ethanol incubated for 240 h. Residual BPA leaching from PC bottles increased with temperature and incubation time. In comparison with the migration observed from PC bottles, non-PC baby bottles and baby bottle liners showed only trace levels of BPA. Tests for leachable lead and cadmium were also conducted on glass baby bottles since these represent a potential alternative to plastic bottles. No detectable lead or cadmium was found to leach from the glass. This study indicated that non-PC plastic baby bottles, baby bottle liners and glass baby bottles might be good alternatives for polycarbonate bottles. PMID- 19680970 TI - Formation of trihalomethanes in foods and beverages. AB - Trihalomethanes (THMs) are suspected carcinogens and reproductive toxicants commonly found in chlorinated drinking water. This study investigates THM formation during the preparation of beverages and foods using chlorinated drinking water. A total of 11 foods and 17 beverages were tested. Under the experimental conditions, each food and beverage formed THMs, primarily chloroform, although low or trace levels of brominated THMs were also detected. Tea formed the highest THM levels (e.g., chloroform levels from 3 to 67 microg l( 1)), followed by coffee (from 3 to 13 microg l(-1)), rice (9 microg l(-1)), soups (from 0.4 to 3.0 microg l(-1)), vegetables (<1 microg l(-1)), and baby food (<0.7 microg l(-1)). Chloroform formation with instant tea, used as a highly reproducible model system, increased with free chlorine concentration, decreased with higher food (tea) concentration, and was unaffected by reaction (steeping) time and bromide ion concentration. These findings indicate that chlorine-food reactions are fast, but that formation decreases as the chlorine demand of the food system increases. THMs are formed in the preparation and cooking of a wide variety of foods if free chlorine is present, and our results suggest that tea can be a significant source of exposure to THMs. PMID- 19680972 TI - Preparation of quality control materials for the determination of sulfonamides in animal feed. AB - A feasibility study of the preparation of quality control materials for the analysis of medicated feeds has been carried out. Two analytical methodologies for the analysis of sulfonamides in feeds were developed, validated and applied to homogeneity and stability studies. Pig feeds spiked with sulfadiazine and sulfadimidine were prepared. The drugs were spiked at 500 microg g(-1), representing what can be expected in a commercial medicated feed, and at 2 and 5 microg g(-1), which roughly correspond to drug-free feeds cross-contaminated during the fabrication process. The homogeneity of both the bulk and the bottled materials was verified. A stability study of the materials containing 2 and 5 microg g(-1) of sulfonamides was carried out over an 18-month period at room temperature, at 4 degrees C and -20 degrees C. The determination of sulfadiazine and sulfadimidine in samples coming from these homogeneity and stability studies of the quality control materials was carried out by high-performance liquid chromatography (HPLC) with either ultraviolet light or fluorimetric detection, depending on the concentration of the analytes in the samples. PMID- 19680973 TI - Evaluation of three different microbial inhibition tests for the detection of sulphamethazine residues in the edible tissues of rabbit. AB - The aim of the present study was to evaluate three microbial inhibition tests (MIT) based on inhibition of growth of the test organisms: (a) four plate test (FPT) containing Bacillus subtilis BGA, (b) screening test for antibiotic residues (STAR) containing Bacillus stearothermophilus var. calidolactis_ATCC 10149 and (c) the Premi(R)Test containing Bacillus stearothermophilus var. calidolactis. The tests were used to determine sulphamethazine (SMZ) residues in edible tissues of rabbit after oral administration up to day 15 of the withdrawal period (WP). A solvent extraction procedure was used to enhance the capability of the tests to detect SMZ residues at or below the maximum residue limit (MRL). Para-aminobenzoic acid (PABA) was employed to previously identify SMZ residues in the first stage of the residue screening. The presence of SMZ residues in the samples was confirmed and quantified by a validated HPLC method. The Premi(R)Test detected SMZ residues in the muscle and heart tissue up to day 9 of the WP, and in the liver, lungs and kidneys up to day 10 of the WP. The STAR detected SMZ residues in the edible organs of rabbits up to day 8 of the WP. The kidneys were positive up to day 5 of the WP, the liver until day 4 of the WP and the lungs until day 3 of the WP. No SMZ residues were detected in the muscle and heart. By using the solvent extraction procedure, SMZ residues were detected in the muscle extract up to day 10 of the WP and the muscle was positive until day 6 of the WP. No detection sensitivity was observed using the FPT. After solvent extraction, SMZ residues were detected in the muscle extract until day 8 of the WP and the muscle was positive until day 3 of the WP. No positive results were detected after the addition of PABA into/onto the agar medium. PABA at a concentration of 10 microg ml(-1) completely reversed the inhibitory activity of SMZ and enabled reliable identification of SMZ in the examined samples. Using HPLC, SMZ was detected in the muscle samples until day 10 of WP (0.02 mg kg(-1)) and in the liver until day 12 of the WP (0.09 mg kg(-1)). The results obtained by the HPLC method and the limit of detection (LOD) of screening tests for SMZ (FPT 0.4 microg ml(-1), STAR 0.2 microg ml(-1), Premi(R) Test 0.05 microg ml(-1)) allowed us to state that the most suitable screening tests for the detection of SMZ residues in the edible tissues of rabbits at level corresponding to the MRL of 0.1 mg kg(-1), established for sulphonamides, are the Premi(R)Test and STAR in conjunction with the solvent-extraction procedure. PMID- 19680971 TI - Estimate of intake of benzoic acid in the Belgian adult population. AB - An exposure assessment was performed to estimate average daily benzoic acid intake for Belgian adults. Food consumption data were retrieved from the national food-consumption survey. As a first step, individual food-consumption data were multiplied with the maximum permitted use levels for benzoic acid per food group (Tier 2). As a second step, a label survey to identify the foods where benzoic acid is effectively used as an additive and a literature review of the possible occurrence of benzoic acid as a natural substance were performed. With this information, a refined list of foods was drafted for the quantification of benzoic acid, which was performed by a high-performance liquid chromatography (HPLC) method, optimized and validated for this purpose. Individual food consumption data were then multiplied with the actual average concentrations of benzoic acid per food group (Tier 3). Usual intakes were calculated using the Nusser method. The mean benzoic acid intake was 1.58 mg kg(-1) body weight day( 1) (Tier 2) and 1.25 mg kg(-1) body weight day(-1) (Tier 3). In Tier 2, men exceeded the acceptable daily intake (ADI) of 5 mg kg(-1) body weight day(-1) at the 99th percentile. The greatest contributors to the benzoic acid intake were soft drinks. Benzoic acid as a natural substance represents only a small percentage of the total intake. The results show that actual benzoic acid intake is very likely to be below the ADI. However, there is a need to collect national food-consumption data for children as they might be more vulnerable to an excessive intake. PMID- 19680974 TI - Predicted intake of trace elements and minerals via household drinking water by 6 year-old children from Krakow (Poland). Part 4: Copper. AB - The aim of this study was to assess exposure of children to copper (Cu) from household drinking water (DW). DW samples were collected between 1997 and 2004 in approximately 650 households and pre-schools using a double-sampling method (morning - W1; evening - W2). The study group comprised approximately 300 children (5-7 years old) living in Krakow (urban, peripheral) and rural areas in southern Poland. Cu concentrations were determined by flame atomic absorption spectrometry. There was no significant relationship between the sampling period and Cu concentration, but statistically significant differences were found between urban and non-urban (rural, peripheral) sites and between morning and evening samples. Geometric means of Cu concentration in evening DW (95% confidence interval, in microg L(-1)) were 8 (1-110), 20 (1-274) and 12 (0-364) in urban, peripheral and rural sites, respectively. DW contamination after overnight standing was comparable in all sites (average increase approximately 20 microg Cu L(-1)). The adopted threshold of 100 microg Cu L(-1) was exceeded in evening DW by 3.6 and 15% in urban and non-urban households, respectively. Exceedance of the EC limit (2 mg L(-1)) was not significant. The mean predicted contribution of evening DW to Cu intake by children were 18-37 microg day(-1) or 0.8-1.6 microg kg(-1) bw day(-1) but were 2-3-fold higher for morning DW. The Polish RDA (1 mg Cu day(-1)) was exceeded for morning (evening) DW in 3.7 (0.2)% of children, with a mean intake of 159 (118)% of RDA. The observed copper levels and predicted intakes can be considered low and should not raise nutritional or toxicological concerns for the age group studied. Nevertheless, due to overnight contamination, the suggestion that the stagnant portion of drinking water should be discarded remains valid. PMID- 19680975 TI - Trace elements in Canadian field peas: a grain safety assurance perspective. AB - Canada is a significant exporter of field peas and, thus, issues of food safety are important in all grain and food products within the international marketplace. Environmental contaminants, such as trace elements, may be present in all foods and, as a result, international standards have been established for a number of toxic trace elements, such as cadmium, lead, mercury and arsenic, in raw food commodities and food products. The Canadian Grain Commission has undertaken a baseline study of various trace elements in Canadian peas to ensure compliance with international food safety legislation. Mean total cadmium content was found to be 0.023 mg kg(-1); arsenic and lead mean values were below the method limit of quantification of 0.050 mg kg(-1) and the total mean mercury level was below the quantification level of 0.002 mg kg(-1). All measured values in the study were below the maximum residue levels (MRLs) established by the FAO and WHO in the Codex Alimentarius. The mean total selenium content was 0.331 mg kg(-1), with 56% of the measured values exceeding the MRL established by the People's Republic of China (PRC) of 0.3 mg kg(-1). No Codex MRL has been established for selenium as it is regarded as an essential trace element for human health and the PRC is currently reviewing its MRL for selenium in light of this fact. For those parts of the world where selenium-deficiency is of nutritional concern, the higher level of selenium in Canadian peas and their products may be of nutritional benefit. PMID- 19680976 TI - Development of an LC-MS/MS method for the determination of pesticides and patulin in apples. AB - A method for the simultaneous determination of 33 pesticides or degradation products together with patulin in apples by liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed. The method involved homogenization of the apples, extraction with ammonium acetate-acetic acid solution in methanol-water by ultrasonication, filtration, and determination by LC-MS/MS. The repeatability and within-laboratory reproducibility for the three spiking levels 0.02, 0.04 and 0.2 mg kg(-1) were between 4% and 35%. In general, the repeatability and reproducibility were about 10-20%. The limits of quantification (LOQs) were between 0.01 and 0.14 mg kg(-1). The method was used on incurred samples from parts of the ISAFRUIT project financed by the European Commission under the 6th Framework Programme. Samples were analysed at four different stages: after harvest, after storage (controlled), after a water bath, and after 28 days at room temperature. Pesticide residues were found at all stages, but no significant differences in the concentration were seen between the stages analysed. The concentration decreased significantly only for tolylfluanid after storage at room temperature for 28 days when only 0-6% of the original amount of tolylfluanid remained in the apples. No patulin was found in the apples stored for 28 days at room temperature and no growth of Penicillium expansum was observed on these apples. However, when the apples were inoculated with a spore suspension of P. expansum, high concentrations of patulin were found. PMID- 19680977 TI - Exposure to pesticides residues from consumption of Italian blood oranges. AB - This paper reports the results of a 5-year study to evaluate pesticide levels, derived from orchard activities, on Italy's most common orange cultivar (Citrus sinensis, L. Osbeck, cv. Tarocco). Using a Bayesian approach, the study allowed both the qualitative (number) and quantitative distributions (amount) of pesticides to be determined with its own probability value. Multi-residue analyses of 460 samples highlighted the presence of ethyl and methyl chlorpyrifos, dicofol, etofenprox, fenazaquin, fenitrothion, imazalil, malathion and metalaxil-m. A total of 30.5% of samples contained just one pesticide, 2.16% two pesticides and 0.65% of samples had three pesticides present simultaneously. The most common residue was ethyl chlorpyrifos followed by methyl chlorpyrifos. Estimated daily intake (EDI) values for ethyl and methyl chlorpyrifos, as well as the distance from the safety level (non-observed adverse effect level, NOAEL), were calculated. The risk was differentiated (1) to take account of the period of actual citrus consumption (180 days) and (2) to discriminate the risk derived from eating oranges containing a certain level of chlorpyrifos from unspecified pesticides. The most likely EDI values for ethyl chlorpyrifos derived from Italian blood orange consumption are 0.01 and 0.006 mg/day calculated for 180 and 365 days, respectively. Considering the probability of the occurrence of ethyl chlorpyrifos, these EDI values are reduced to 2.6 x 10(-3) and 1.3 x 10(-3) mg/day, respectively. For methyl chlorpyrifos, the most likely EDI values are 0.09 and 0.04 mg/day, respectively; considering the probability of its occurrence, the EDI values decrease to 6.7 x 10(-3) and 3.4 x 10(-3) mg/day, respectively. The results confirmed that levels of pesticides in Italian Tarocco oranges derived from a known controlled chain of production are safe. PMID- 19680978 TI - Influence of post-harvest application rates of cyprodinil, treatment time and temperature on residue levels and efficacy in controlling green mould on 'Valencia' oranges. AB - The effectiveness of heat treatments with water and cyprodinil in controlling post-harvest green mould caused by Penicillium digitatum was investigated on artificially inoculated 'Valencia' oranges. Residue levels of cyprodinil were determined in the oranges as a function of active ingredient concentration, temperature and treatment time. Cyprodinil residues were significantly dependent on treatment time when applied at 600 mg l(-1) and 20 degrees C, but not when fruit were treated at 150-300 mg l(-1). The application of cyprodinil at 50 or 100 mg l(-1) at 55 degrees C for 30 s produced similar residue levels, while residues increased when the application rate was 150 mg l(-1). Cyprodinil at 100 mg l(-1) and 60 degrees C produced a significant increase in residues compared to treatment at 50 mg l(-1); no significant increase in residues was found when the application rate was raised from 100 to 150 mg l(-1). In comparison to treatments performed at 20 degrees C, the application of a heated cyprodinil mixture resulted in significantly higher residues in fruit. All treatments with cyprodinil at 20 degrees C similarly reduced green mould after 7 days of storage at 20 degrees C. After 18 days, treatment with cyprodinil at 600 mg l(-1) for 30 s was more effective than at 150-300 mg l(-1). When dip time was extended to 90 or 180 s, treatment efficacy was positively related to fungicide concentration. Treatments with water at 55 degrees C for 30 s were as effective as cyprodinil at 50-100 mg l(-1), but less effective than cyprodinil at 150 mg l(-1). After 7 days, treatment with water or cyprodinil at 50-150 mg l(-1) and 60 degrees C were equally effective in controlling green mould; while, after 18 days, treatment with cyprodinil at 150 mg l(-1) was consistently more effective than at 50-100 mg l(-1) or hot water alone. PMID- 19680979 TI - Probabilistic assessment of the cumulative dietary acute exposure of the population of Denmark to organophosphorus and carbamate pesticides. AB - Organophosphorus and carbamate pesticides are acetylcholinesterase-inhibiting pesticides and as such have a common mode of action. We assessed the cumulative acute exposure of the population of Denmark to 25 organophosphorus and carbamate pesticide residues from the consumption of fruit, vegetables and cereals. The probabilistic approach was used in the assessments. Residue data obtained from the Danish monitoring programme carried out in the period 2004-2007, which included 6704 samples of fruit, vegetables and cereals, were used in the calculations. Food consumption data were obtained from the nationwide dietary survey conducted in 2000-2002. Contributions from 43 commodities were included in the calculations. We used the relative potency factor (RPF) approach to normalize the toxicity of the various organophosphorus and carbamate pesticides to the two index compounds chlorpyriphos and methamidophos. RPF values derived from the literature were used in the calculations. We calculated the cumulative acute exposure to 1.8% and 0.8% of the acute reference dose (ARfD) of 100 microg kg(-1) body weight (bw) day(-1) of chlorpyrifos as an index compound at the 99.9th percentile (P99.5) for children and adults, respectively. When we used methamidophos as the index compound, the cumulative acute intakes were calculated to 31.3% and 13.8% of the ARfD of 3 microg kg(-1) bw day(-1) at P99.9 for children and adults, respectively. With both index compounds, the greatest contributor to the cumulative acute exposure was apple. The results show that there is no cumulative acute risk for Danish consumers to acetylcholinesterase inhibiting pesticides. PMID- 19680981 TI - Fusarium mycotoxin content of UK organic and conventional oats. AB - Every year between 2002 and 2005 approximately 100 samples of oats from fields of known agronomy were analysed by GC/MS for 10 trichothecenes: deoxynivalenol (DON), nivalenol, 3-acetylDON, 15-acetylDON, fusarenone X, T-2 toxin (T2), HT-2 toxin (HT2), diacetoxyscirpenol, neosolaniol and T-2 triol. Samples were also analysed for moniliformin and zearalenone by HPLC. Of the 10 trichothecenes analysed from 458 harvest samples of oat only three, 15-acetylDON, fusarenone X and diacetoxyscirpenol, were not detected. Moniliformin and zearalenone were absent or rarely detected, respectively. HT2 and T2 were the most frequently detected fusarium mycotoxins, present above the limit of quantification (10 microg kg(-1)) in 92 and 84% of samples, respectively, and were usually present at the highest concentrations. The combined mean and median for HT2 and T2 (HT2 + T2) was 570 and 213 microg kg(-1), respectively. There were good correlations between concentrations of HT2 and all other type A trichothecenes detected (T2, T2 triol and neosolaniol). Year and region had a significant effect on HT2 + T2 concentration. There was also a highly significant difference between HT2 + T2 content in organic and conventional samples, with the predicted mean for organic samples five times lower than that of conventional samples. This is the largest difference reported for any mycotoxin level in organic and conventional cereals. No samples exceeded the legal limits for DON or zearalenone in oats intended for human consumption. Legislative limits for HT2 and T2 are currently under consideration by the European Commission. Depending on the limits set for unprocessed oats intended for human consumption, the levels detected here could have serious consequences for the UK oat-processing industry. PMID- 19680980 TI - Antimutagenic and antigenotoxic effects of vegetable matrices on the activity of pesticides. AB - Two in vitro tests, one to detect bacterial mutagenicity (Ames test) on Salmonella typhimurium TA98, TA100, and TA1535 and the other the primary DNA damage (SOS Chromotest) on Escherichia coli PQ37, were applied to determine the overall genotoxic activity of 12 pesticides (azinphos methyl, chlorothalonil, chlorphyriphos ethyl, chlorphyriphos methyl, lambda-cyhalothrin, cypermethrin, cyprodinil, fenazaquin, fludioxonil, indoxacarb, iprodione and penconazol). These were detected by gas chromatography (GC) analysis with electron capture (ECD) and nitrogen phosphorus detection (NPD) in 18 samples of vegetables. Some extracts of vegetables, found positive for pesticides with GC, were subjected to the Ames test and SOS Chromotest to evaluate the possible antimutagenic and/or antigenotoxic effects of vegetable matrices. The same bioassays were also performed on the mixtures of pesticides found in these samples to evaluate whether interactions could occur between pesticides and be responsible for the possible antimutagenic and/or antigenotoxic effects of the contaminated matrices. Experiments were also carried out to compare the results found for contaminated vegetables with their content of antioxidant components. Significant differences in mutagenicity and genotoxicity were found among the pesticides selected for this study. Of the 12 pesticides tested, only azinphos methyl, cyprodinil, fludioxonil and iprodione were found to be positive for both S. typhimurium and E. coli. No mutagenic/genotoxic activity was found in the extracts of vegetables contaminated by pesticides. S. typhimurium TA1535 showed a strong positive mutagenic effect for the mixtures of pesticides while they were not able to induce the SOS system. The data concerning the content of polyphenols and the total reducing activity of the contaminated vegetables indicated high amounts of antioxidants that could explain the inhibitory effect on the activity of pesticides shown by vegetables. PMID- 19680982 TI - Deoxynivalenol suppresses circulating and splenic leukocyte subpopulations in BALB/c mice: dose response, time course and sex differences. AB - It was hypothesized that suppression of peripheral blood leukocyte subsets were markers of exposure to dietary deoxynivalenol (DON), a Fusarium graminearum mycotoxin in grain, at 1.0 mg kg(-1) but not at lesser doses in BALB/c mice. Groups of 10 female and 10 male BALB/c mice were fed 0, 0.25, 0.5, 1.0, and 2.0 mg kg(-1) DON for 14 and 28 days. Using flow cytometry with staining for leukocyte surface markers, the percentage of CD19(+) leukocytes (B cells) in peripheral blood was decreased in both sexes of BALB/c mice after 14 days of exposure to 1.0 or 2.0 mg kg(-1) DON, whereas exposure to DON over 28 days did not inhibit B cells compared to the control diet. The percentage of mononuclear cells in peripheral blood was decreased in female BALB/c mice fed 1 and 2 mg kg( 1) DON after 14 days compared with control diet. The percentage of CD11b(+) leukocytes (monocytes) in peripheral blood and total CD11b(+) splenic leukocytes were decreased only in female mice fed 1.0 and 2.0 mg kg(-1) DON after 28 days compared with control diet, which shows the greater sensitivity to DON in females compared to males. It was concluded that BALB/c mice adapted to DON exposure because peripheral blood cellular effects of DON at 14 days disappeared by 28 days with the exception of monocyte changes in females. This suggests that female sex hormones potentiate one potential marker of DON immunotoxicity in BALB/c mice. PMID- 19680983 TI - Impact of industrial treatments on ochratoxin A content in artificially contaminated cocoa beans. AB - Ochratoxin A (OTA) is a mycotoxin mainly produced by mould species of the genera Aspergillus and Penicillium, which grow on a variety of agricultural products. OTA-contaminated foodstuffs pose a major health hazard to consumers, including human and animal. In Cote d'Ivoire, numerous studies are being carried out to find the best way of preventing OTA contamination of cocoa raw material. The objectives of this investigation were to assess the impact of industrial treatment on OTA content in cocoa-derived products. Samples of cocoa pods were prepared under specific conditions promoting fungal proliferation on cocoa beans before processing. The beans underwent the usual industrial treatments - roasting, shelling, crushing, pressing and additive addition - and samples were taken at each stage. OTA was extracted with a methanol/3% sodium hydrogen carbonate solution and purified using an immunoaffinity column prior to HPLC analysis with fluorescence detection. OTA was detected in artificially contaminated cocoa beans at levels ranging from 3.4 to 44.7 microg kg(-1) with a mean value of 22.9 +/- 3.6 microg kg(-1). OTA was mainly concentrated in the shell (93%). Roasting, shelling and additive addition significantly decreased levels of OTA by 24-40, 76 and 52%, respectively, with an overall reduction of approximately 91%. These results indicate that industrial processing of cocoa has a real impact on the reduction of OTA in final cocoa products. PMID- 19680984 TI - Effect of some habitual cooking processes on the domoic acid concentration in the cockle (Cerastoderma edule) and Manila clam (Ruditapes philippinarum). AB - The effect of cooking on the concentration and burden of domoic acid in two bivalve molluscs was studied. The Manila clam (Ruditapes philippinarum) and cockle (Cerastoderma edule) were subjected to steaming and boiling, respectively. In both cases, factorial plans were used to evaluate the effects of common cooking methods and the variations likely to take place during the cooking procedure (cooking time and salt concentration in both species, in addition to ethanol percentage in Manila clam). The domoic acid concentration and toxin content were affected by cooking in very different ways in the two species studied. The cockle lost a significant part of its domoic acid content, while the clam did not. Since the weight of the soft tissues in cooked bivalves was lower than in the raw samples in both species, the toxin concentration decreased less than the toxin burden in the cockle, while it increased in the clam, where the toxin burden did not change significantly. Among the cooking variables tested, only cooking time had a noticeable effect on the domoic acid content in the clam and cockle, with the bivalves that were cooked for a longer time having smaller amounts of toxin. It is clear that cooking affects the toxin concentration in bivalves in a way that is species specific. This characteristic must be taken into account when evaluating epidemiological information, establishing allowable toxin levels and in cases where pre-processing treatments such as cooking or similar methods are used in monitoring systems. PMID- 19680985 TI - Determination of polycyclic aromatic hydrocarbons (PAHs) in commonly consumed Nigerian smoked/grilled fish and meat. AB - Smoking and/or grilling, when carried out with traditional methods involving direct contact with wood combustion fumes, is responsible for high contamination levels with carcinogenic polycyclic aromatic hydrocarbons (PAHs). The aim of this work was to investigate the PAH content of different smoked or grilled meat and fish products commonly consumed in Nigeria. A rapid method involving microwave assisted saponification and simultaneous extraction followed by solid-phase extraction (SPE), high-performance liquid chromatography (HPLC) separation and spectrofluorometric detection was employed. Samples that were smoked or grilled using traditional systems, which use a wood fire, were heavily contaminated with benzo[a]pyrene (BaP) at levels ranging from 2.4 to 31.2 microg kg(-1) wet weight. Considerably lower contamination levels were found in samples smoked or grilled in the laboratory using a charcoal fire (BaP from 0.7 to 2.8 microg kg(-1) wet weight). The health risk associated with a daily consumption of 100 g of these products was also evaluated using the margin of exposure (MOE) approach. MOE values lower than 10,000 were obtained for all smoked/grilled commercial samples, indicating a potential concern for consumer health. PMID- 19680986 TI - Effect-based and chemical analysis of polycyclic aromatic hydrocarbons in smoked meat: a practical food-monitoring approach. AB - Polycyclic aromatic hydrocarbons (PAHs), which are generated by heat treatment and smoke curing of meat, pose a risk to human health. At present, the determination of these unwanted contaminants requires costly, time-consuming chemical analysis of smoked meat. An alternative is effect-directed high throughput bioassays, which could also be used as a pre-screening method. The authors recently adapted the in vitro chemical-activated luciferase gene expression (CALUX) assay as a rapid, sensitive, and inexpensive screening technique for compounds such as dioxins, polychlorinated biphenyls, and PAHs. The aim of the present study was to apply a practical approach under realistic conditions. Custom-made meat samples produced under defined conditions with different PAH levels were analysed using this bioassay and gas chromatography mass spectrometry (GC/MS) to determine the influence of different smoking conditions (temperature and duration) on PAH levels. It was found that cold smoking for up to 6 h did not result in strong PAH contamination, whereas hot (65 degrees C) and longer smoking times caused a considerable increase in both the bioassay response and the levels of 31 individually determined PAHs. The response in the effect-based bioassay was in good agreement with the values of chemical analysis. The bioassay made it possible to determine accurately the degree of contamination. The results show that this assay is suitable for high-throughput screening for unknown levels of toxicologically relevant PAHs in meat samples and is sensitive enough to differentiate between different PAH levels generated under various smoking conditions. Effect-based screening techniques, therefore, provide a new instrument for official food monitoring. PMID- 19680987 TI - Search for a more adequate test to predict the long-term migration from the PVC gaskets of metal lids into oily foods in glass jars. AB - As shown previously, the conventional testing procedure for simulating long-term migration from the gaskets of metal closures into oily foods does not adequately reflect reality. It appears to be impossible to accelerate migration to the extent that the situation at the end of the shelf life of a product can be anticipated in a few days or weeks. Therefore, we investigated whether long-term migration could be extrapolated from migration rates determined for new lids. Jars were kept in the normal upright position. Since heat treatment may have a strong temporary impact, migration during the initial heating for pasteurization or sterilization and storage at ambient temperature were determined using different lids. Commercial products were recalled from sales points throughout Europe to determine the real migration over extended periods of time and for jars with differing histories. This migration was compared with data from the short term testing to investigate whether an empirical relationship could be derived. The results show that the short-term test enables the comparison of lids and plasticizers in the initial phase of migration, but that long-term extrapolation presupposes more complex kinetic modeling. The results also demonstrate that the legal relevance of "official" testing methods should be reconsidered to avoid conflict when food contact materials comply with migration limits in the test but not in actual application. PMID- 19680988 TI - Long-term functional recovery in the rat auditory system after unilateral auditory cortex ablation. AB - CONCLUSIONS: An intact bilateral auditory corticofugal projection is necessary for the auditory system, and above all its main targets, to start working correctly after an acoustic stimulation. After restricted unilateral cortical lesions, the auditory system is able to recover its function in adult animals at 90 days after surgery. (Post-lesion plasticity in adults.) OBJECTIVES: To study the influence of the cortex on the auditory system functionality and to asses its ability for post-lesion recovery. METHODS: Restricted unilateral lesions were made in the auditory cortex of adult rats. To evaluate the functionality of the auditory pathway after corticofugal deafferentation, the acoustic startle reflex and prepulse inhibition, together with the auditory brainstem response (ABR), were tested along the survival time. RESULTS: All the three tests showed a decrease in their responses at 15 days post lesion, and a full recovery at 90 days post lesion for the ABR and at 180 days post lesion for the acoustic startle reflex and prepulse inhibition. PMID- 19680989 TI - Basic fibroblast growth factor combined with biodegradable hydrogel promotes healing of facial nerve after compression injury: an experimental study. AB - CONCLUSION: Topical application of basic fibroblast growth factor (bFGF) hydrogel facilitates faster healing from traumatic facial paralysis due to continuous release of bFGF. OBJECTIVES: bFGF is considered a potent agent to facilitate recovery from neuronal damage; however, exogenously applied bFGF does not work well because of its short acting time. To enhance the effects in vivo, we developed a new drug delivery system by embedding bFGF in a gelatin hydrogel that degrades slowly. In this study, the effects of bFGF-hydrogel on traumatic facial nerve paralysis were investigated in guinea pigs. METHODS: The intratemporal facial nerve was exposed and clamped at the vertical portion using micro needle forceps. The animals were then subjected to one of the following three procedures: group A, no further treatment; group B, one-shot application of bFGF to the nerve; and group C, application of bFGF-hydrogel instead. Six weeks later, facial nerve functions were evaluated by three test batteries: observation of facial movements, electrophysiological testing, and histological study. RESULTS: The results for groups A and B were similar in the three tests, indicating that one-shot application of bFGF did not benefit facial nerve recovery. In contrast, group C achieved better results in all tests. PMID- 19680990 TI - Human papillomavirus (HPV)-positive tonsillar carcinomas are frequent and have a favourable prognosis in males in Norway. AB - CONCLUSIONS: This study confirms a high prevalence of human papillomavirus (HPV) positive tonsillar tumours (52%). The survival of the HPV-positive group was significantly better in males. OBJECTIVES: We assessed the prevalence of HPV in 137 patients with tonsillar carcinomas, measured the p53- and Ki-67-positive tumour cell fractions and correlated the results with clinical variables. PATIENTS AND METHODS: Tumour DNA from patients with squamous cell carcinoma of the tonsillar region was amplified by PCR and sequenced for detection of HPV subtypes. RESULTS: HPV was found in 71/137 (52%) of the tumours; HPV-16 was the most frequent subtype (87%). HPV positivity did not correlate with gender, stage, T- and N categories, Ki-67 expression or p53 positivity. The HPV-positive group had a significantly better survival (p < 0.01) compared with the HPV-negative group in males. In a multivariate analysis HPV status gave prognostic information in addition to the earlier established factors, i.e. age, gender and stage (p < 0.05). PMID- 19680991 TI - The effects of age at cochlear implantation and hearing aid trial on auditory performance of Chinese infants. AB - CONCLUSION: Most of the infants demonstrated rapid improvement in the three different auditory skills within the first year after switch-on. Infants undergoing hearing aid trial and habilitation demonstrated a significant positive effect on the development of auditory skills in comparison with infants without trial and habilitation. OBJECTIVE: This paper aims to evaluate the auditory performance of infants of different age at cochlear implantation, emphasize the importance of the hearing aid trial and habilitation before implant, and provide baseline data of auditory development. METHODS: In all, 259 infants with prelingually profound hearing loss participated in this study. The Infant-Toddler Meaningful Auditory Integration Scale (IT-MAIS) was used to assess auditory skills in infants at different intervals. RESULTS: The mean scores for the auditory skills improved significantly over time. The mean scores of three skills for each group at each interval were significantly different. The mean scores of the auditory skills were significantly superior for infants undergoing hearing aid trial and habilitation in comparison with those of infants without hearing aid trial and habilitation. PMID- 19680996 TI - Dehydroepiandrosterone increased oxidative stress in a human cell line during differentiation. AB - Dehydroepiandrosterone (DHEA), a reversible inhibitor of glucose-6-phosphate dehydrogenase (G6PD), is increasingly taken as an antioxidative and anti-ageing supplement. This study investigated the effects of DHEA on the expression of G6PD and on the state of oxidative stress in a human promyelocytic leukaemia cell line, HL60, during the differentiation to neutrophil-like cell. This study differentiated HL60 with dimethyl sulfoxide (DMSO) in the presence (DMSO HL60/DHEA) or absence (DMSO-HL60) of DHEA. During the differentiation, activity, mRNA and protein levels of G6PD were increased. DHEA increased these levels further. DHEA by itself suppressed the production of superoxide from DMSO-HL60 upon stimulation with phorbol myristate acetate (PMA). However, DMSO-HL60/DHEA stimulated with PMA in the absence of DHEA produced superoxide and 8-oxo deoxyguanosine more than PMA-stimulated DMSO-HL60. After addition of H(2)O(2), the ratio of reduced glutathione to oxidized glutathione was lower in DMSO HL60/DHEA than in DMSO-HL60. These findings indicate that DHEA acts both as an antioxidant and as a pro-oxidant. PMID- 19680997 TI - Pitfalls in the use of arachidonic acid oxidation products to assign lipoxygenase activity in cancer cells. AB - Arachidonic acid (AA) reaction with cyclooxygenase (COX) and lipoxygenases (LOX) yield eicosanoids that can mediate prostate cancer proliferation and enhance both tumour vascularization and metastasis. Increasingly measurement of eicosanoids with liquid chromatography is employed to implicate LOX activity in different biological systems and in particular link LOX activity to the progression of cancer in experimental models. This study demonstrates that simply identifying patterns of eicosanoid regio-isomerism is insufficient to designate LOX activity in prostate cancer cells and the analysis must include complete stereochemical assignment of the various isomers in order to validate the assignment of LOX activity. PMID- 19680998 TI - Induction of heat shock proteins prevents the arrhythmogenic substrate for atrial fibrillation. AB - Atrial fibrillation (AF) is the commonest arrhythmia. Studies have shown that atrial tachypacing (artificial persistent AF) causes electrical remodelling. This is characterised by the shortening of the atrial effective refractory period (ERP), in which reduction in L-type Ca(2+) channel current plays an essential part. Atrial fibrosis, a feature of structural remodelling, is induced by continuous infusion of angiotensin II, and has been associated with conduction delay in atria, which promotes AF. Acute atrial ischaemia, frequently observed during development of acute coronary syndrome, has been associated with atrial conduction heterogeneity, which also promotes AF. Induction of heat shock proteins (Hsp72 and Hsp27) by hyperthermia and/or geranylgeranylacetone has demonstrated to protect the heart against such atrial remodelling. The potent protective role of Hsp72 and Hsp27 against clinical AF in patients who underwent open heart surgery has been shown. Taken together, interventions that induce heat shock responses (including induction of Hsp72 and Hsp27) may prevent newly developed AF and delay the progression of paroxysmal AF to persistent AF. PMID- 19681000 TI - Understanding mealtime changes for adults with cerebral palsy and the implications for support services. AB - BACKGROUND: Changes in the swallowing capabilities of adults with cerebral palsy as they age may impact on their health, safety, and well-being. METHOD: Thirty two adults with cerebral palsy aged between 30 and 69 years participated in in depth interviews about their experiences of changes in their swallowing and related management of their mealtimes within the last two years. A constant comparative qualitative analysis of the interviews elucidated the changes they experienced. RESULTS: Changes included increased coughing and choking, digestive or gastro-oesophageal symptoms, diet modification, loss of independence with psychosocial consequences. Participants reported unsatisfactory collaboration with service providers over mealtime management decisions and interventions. CONCLUSIONS: Adults with cerebral palsy may experience gradual changes in their swallowing and mealtime capabilities from as early as 30 years of age. Regular collaborative assessment and involvement of all stakeholders in decisions are important to facilitate compliance with recommendations, ongoing safety, and optimal well-being. PMID- 19681001 TI - Meta-analysis of the effectiveness of individual intervention in the controlled multisensory environment (Snoezelen) for individuals with intellectual disability. AB - BACKGROUND: The Snoezelen is a multisensory intervention approach that has been implemented with various populations. Due to an almost complete absence of rigorous research in this field, the confirmation of this approach as an effective therapeutic intervention is warranted. METHOD: To evaluate the therapeutic influence of the Snoezelen approach. Twenty-eight relevant articles relating to individual (one-to-one) Snoezelen intervention with individuals with intellectual and developmental disabilities (IDD) were reviewed. A meta-analysis regarding the significance of the reduction of maladaptive behaviour and the enhancement of adaptive behaviour was implemented. An analysis of standardised mean differences was used through the use of fixed effect models. RESULTS: The primary finding was that the Snoezelen approach, when applied as an individual intervention for individuals with IDD, enabled significant and large effect size in adaptive behaviours, with generalisation to the participants' daily life. CONCLUSIONS: Weaknesses in the examined research methodologies, the heterogeneity between research designs, the small number of available research projects, and the small number of participants in each research project, prevent a confirmation of this method as a valid therapeutic intervention at this time. PMID- 19681002 TI - Down or up? Explaining positive and negative emotions in parents of children with Down's syndrome: Goals, cognitive coping, and resources. AB - BACKGROUND: Some studies find that parents of children with Down's syndrome may experience symptoms of depression, while other studies find that parents adapt well. This study aimed to clarify this variability in adaptive strength by investigating a stress-coping model to explain depressive symptoms and positive affect. METHOD: Questionnaires were completed by 553 parents of children (aged 0 18) with Down's syndrome, containing measures of goal disturbance, cognitive coping, social support, partner bonding, and coping self-efficacy. RESULTS: Different models for positive affect and depressive symptoms were found. The coping strategies of self-blame and rumination were positively related to depressive symptoms, and positive reappraisal was positively related to positive affect. Partner bonding characteristics played relevant roles in both models, as did coping self-efficacy and goal disturbance. Social support seemed mainly relevant in explaining positive affect. CONCLUSIONS: Different psychological factors were related to depressive symptoms and positive affect in parents of children with Down's syndrome. Implications are discussed. PMID- 19681003 TI - Standardised Observation Analogue Procedure (SOAP) for assessing parent and child behaviours in clinical trials. AB - BACKGROUND: Observational measures of parent and child behaviours have a long history in child psychiatric and psychological intervention research, including the field of autism and developmental disability. We describe the development of the Standardised Observational Analogue Procedure (SOAP) for the assessment of parent-child behaviour before and after a structured parent training program for children with pervasive developmental disorders (PDD). We report on the use of this procedure in a pilot study of 12 participants with PDD. RESULTS: Inter-rater reliability across behaviours coded ranged from 75-100% agreement. Blindly scored observations of behaviour showed medium effect sizes for changes in inappropriate child behaviour. Analyses of baseline scores revealed a moderate positive correlation between inappropriate child behaviours as measured in all four SOAP conditions and parent ratings of child noncompliance (r(s) = .66, p < .05). By contrast, the correlations of SOAP scores with parent ratings of irritability was lower (r(s) = .40, p >.05). CONCLUSIONS: As our treatment targeted compliance, these preliminary results suggest that the SOAP provides a valid measure of noncompliant behaviour in children with PDD and is sensitive to treatment effects on inappropriate child behaviours. PMID- 19681004 TI - Service and support needs of Australian carers supporting a family member with disability at home. AB - BACKGROUND: As part of an international, multicentre project, the service and support needs of Australian family carers were investigated. METHOD: A sample of 1,390, 448 family carers completed a self-report survey, including an adaptation of the Family Needs Survey (FNS) and several open-ended questions. A mixed method design was used, employing quantitative and qualitative analyses. RESULTS: On the FNS the most frequently endorsed items were those relating to the need for information about services and, in particular, future, out-of-home accommodation. Similarly, the need for respite services was endorsed by over 80% of respondents. Comments indicated that access to and the quality of respite, day support, and therapy programs were a priority. CONCLUSIONS: Participants expressed the need for greater access to information. Access to appropriate respite options, together with quality day support and therapy services, remain a priority for family carers. PMID- 19681005 TI - Transitions from school for young adults with intellectual disability: Parental perspectives on "life as an adjustment". AB - BACKGROUND: Few studies have investigated transition programs and outcomes for young adults with disabilities as viewed from the parent perspective. The current Australian study provided a voice for parents to report on the experiences of and outcomes for young adults following their recent transition from school into post school life. METHOD: A mailed survey gathered statewide data from parents (N = 218) whose family member had completed schooling at a secondary or special school. The majority of those participating were parents of adults with intellectual disability and high support needs. RESULTS: Findings detail school preparation for employment, community activities, and daily living; parent and student involvement in transition goal-setting; and post-school outcomes for young adults and their families. CONCLUSIONS: Parental reflections on family adjustment and life satisfaction for the young adult since leaving school highlight the lack of post-school options and the impact of these circumstances on the whole family. PMID- 19681006 TI - A content analysis from a US statewide survey of memorable healthcare decisions for individuals with intellectual disability. AB - BACKGROUND: Little is known about surrogate healthcare decision-making for individuals with intellectual disability (ID). This study examined healthcare decision-making by residential-agency directors to learn their process and the extent to which the individual is included. METHOD: Content analysis of qualitative data from a mailed survey of residential-agency directors in a large US northeastern state. RESULTS: Narrative comments of 102 directors (65% of respondents) are reported. Three themes emerged: (a) Identifying someone else's "best interest" is challenging; (b) Perceptions of the healthcare community, especially related to quality of life, can influence care provided; and (c) Surrogate decision-making is a team effort. CONCLUSIONS: With knowledge of how decisions are made, the healthcare community can better interact with the complex array of service agencies and persons who determine care for this vulnerable population. PMID- 19681007 TI - The Kirkpatrick model: A useful tool for evaluating training outcomes. AB - BACKGROUND: Services employing staff to support people with disability usually provide training in a range of areas including communication and managing challenging behaviour. Given that such training can be costly and time-consuming, it is important to evaluate the evidence presented in support of such programs. Efficacy in clinical practice is measured using evidence-based practice. However, there is currently no model that is widely used to compare and evaluate training programs despite the large number of training programs reported each year. METHOD: Six studies published in the last decade that reported the outcomes of communication-based training and six that reported on the outcomes of challenging behaviour training were evaluated using the 4-level Kirkpatrick model. RESULTS: Comparison of the levels of evidence is made for these 12 studies. CONCLUSION: The Kirkpatrick model provides one technique for appraisal of the evidence for any reported training program and could be used to evaluate whether a training program is likely to meet the needs and requirements of both the organisation implementing the training and the staff who will participate. PMID- 19681011 TI - Preparing for the worst. PMID- 19681008 TI - Extrapyramidal symptoms and medication use in Mucopolysaccharidosis type III. AB - BACKGROUND: We report the case of a 16-year-old male with Mucopolysaccharidosis III type A (Sanfilippo syndrome) who was commenced on risperidone for behaviour management. He rapidly developed extrapyramidal symptoms that have not resolved. METHOD: The medication histories of 20 patients with Mucopolysaccharidosis III seen at a Lysosomal Storage Diseases Clinic were reviewed to determine the incidence of extrapyramidal side effects. RESULTS: Six patients had been treated with risperidone, olanzapine, or lamotrigine. Five of these patients developed extrapyramidal side effects. CONCLUSIONS: The incidence of extrapyramidal side effects was considerably higher than expected. We suggest that these medications be used with considerable caution in these patients. PMID- 19681010 TI - State codification of federal regulatory ambiguities in biobanking and genetic research. PMID- 19681012 TI - Legal remedies for online defamation of physicians. PMID- 19681013 TI - A physician's drug use and the duty to disclose provider problems. PMID- 19681014 TI - ERISA preemption and the question of pharmacy benefit managers' fiduciary duty. PMID- 19681017 TI - Radial deformation by tissue Doppler imaging in multiple myocardial layers. AB - OBJECTIVES: Measuring strain in multiple myocardial layers using 2-dimensional tissue Doppler imaging may provide valuable diagnostic information about non transmural disease. However, its feasibility in humans has not been demonstrated previously, and optimal machine settings not defined. DESIGN: From parasternal short axis, zoomed tissue Doppler imaging of the left ventricular inferior wall was obtained in 23 young, healthy humans. Images were recorded with six different machine settings. Velocity, strain rate and strain were measured in two and three layers across the wall. RESULT: For two-layer-measurements, all subendocardial values were significantly higher than the subepicardial ones (p<0.003 for all data sets). Minimal radial and maximal lateral averaging resulted in largest strain differences: 106.4+/-32.6% (mean+/-SD) subendocardially versus 54.2+/ 20.1% subepicardially. By similar settings in three layers, strain was 105.7+/ 34.5% subendocardially, 81.2+/-26.9% midmyocardially and 48.3+/-26.9% subepicardially (p <0.05). CONCLUSION: We have demonstrated that it is feasible to measure radial velocity, strain rate and strain in up to three individual layers in young healthy humans, and the diagnostic potentials should be tested on patient groups. PMID- 19681018 TI - Peripheral neuropathy in inflammatory bowel disease patients: a prospective cohort study. PMID- 19681019 TI - Empiric antimicrobial therapy of community-acquired pneumonia: clinical diagnosis versus procalcitonin levels. PMID- 19681020 TI - Prevalence of ST26 among untreated smear-positive tuberculosis patients from Karachi indicating ongoing transmission. AB - Tuberculosis (TB) control is a major healthcare priority for Pakistan. We have studied Mycobacterium tuberculosis strains from the sputa of 100 treatment-naive, smear-positive pulmonary TB cases from Karachi, Pakistan, to identify strains most responsible for active transmission in this population. DNA extracted from M. tuberculosis isolates were subjected to spacer oligotyping (spoligotyping). Sixty-six (66%) clinical isolates were grouped into 9 different clusters. The largest cluster comprised the Central Asian Strain (CAS) 1 or ST26 (n=40). The remaining isolates (34%) had unique spoligotypes. We conclude that ST26 being the most prevalent strain in smear-positive cases contributes greatly towards ongoing transmission in Karachi. Our data further suggest that ST26 may have a selection advantage not afforded by other genotypes. This conclusion is further supported by DESTUS analysis (Detecting Emerging Strains of Tuberculosis Using Spoligotypes) identifying ST26 as the only emerging spoligotype. Reasons for the spread of ST26 require further study. PMID- 19681021 TI - Community-acquired complicated intra-abdominal infections in children hospitalized during 1995-2004 at a paediatric surgery department. AB - Information on the epidemiologic, clinical, microbiologic and therapeutic aspects of community-acquired complicated intra-abdominal infections in paediatrics is limited. The objectives of this study were to investigate the epidemiologic, clinical, microbiologic and therapeutic characteristics of community-acquired complicated intra-abdominal infections occurring in children aged 1 month-15 y. Medical charts and microbiology data of all children hospitalized with complicated intra-abdominal infections were retrospectively examined. Complicated intra-abdominal infections were defined as infections extending beyond the viscus of origin into the peritoneal space, with subsequent development of abscess or peritonitis. One hundred and twenty-three patients with complicated intra abdominal infections (99/123, 80% >5 y of age) were included; 113 (92%) had complicated acute appendicitis. Twelve (10%) patients underwent computerized tomography-guided percutaneous drainage of periappendicular abscesses. The mean rate of complicated intra-abdominal infections among patients with complicated acute appendicitis was 10% without significant changes during the study years. Positive intra-abdominal cultures were recorded in 97/108 (90%) evaluable patients; 65/97 (67%) cases were characterized by mixed bacterial flora growth. One hundred and ninety pathogens (aerobes n=164, 86%; anaerobes n=26, 14%) were isolated. Escherichia coli was the most common pathogen (94 isolates, 57%). In vitro amoxicillin/clavulanate coverage of E. coli and Klebsiella spp was modest (81% and 86%, respectively). The ampicillin/gentamicin/metronidazole regimen was more appropriate in vitro than the amoxicillin/clavulanate regimen (3/80, 4% resistant pathogens compared with 8/43, 19%; p=0.02). Post-operative complications were recorded in 33/123 (27%) patients. Time until defervescence to < 37.5 degrees C was shorter in children with periappendicular abscess than in children with generalized peritonitis (6+/-4 vs 4+/-3 days; p=0.009). IN CONCLUSION: (1) most community-acquired complicated intra-abdominal infections occurred as a result of acute appendicitis; (2) the rate of complicated intra abdominal infections among patients with acute appendicitis was low and without significant changes during the study period; (3) E. coli was the most frequently isolated pathogen; (4) amoxicillin/clavulanate provided only partial coverage for complicated intra-abdominal infection pathogens and should be used with caution in the empiric treatment of complicated intra-abdominal infections. PMID- 19681023 TI - Indications for the use of endoscopic mucosal resection for early gastric cancer in Japan: a comparative study with endoscopic submucosal dissection. AB - BACKGROUND AND STUDY AIMS: Endoscopic submucosal dissection (ESD) has been reported to produce excellent treatment results for early gastric cancer. In terms of lesions that previously met the criteria for endoscopic mucosal resection (EMR), there is now controversy about which of the two methods is superior, and whether the two methods are comparable. PATIENTS AND METHODS: A total of 177 patients (202 lesions) with early gastric cancer who met the guidelines for EMR and who underwent either EMR or ESD were studied. The rates of en bloc resection, complete resection, local recurrence, and complications were compared between EMR and ESD. RESULTS: The overall en bloc and complete resection rates were lower in patients undergoing EMR than in those undergoing ESD (en bloc: 53.8 % vs. 94.3 %, P < 0.001; complete: 37.5 % vs. 92.6 %, P < 0.001). The overall 5-year recurrence-free rate was lower in the EMR group than in the ESD group (82.5 % vs. 100 %; P < 0.001). However, with regard to the tumor size, the two groups did not differ in en bloc ( P = 1.0) or complete resection rate ( P = 0.8) for tumors < or = 5 mm and in 5-year recurrence-free rate ( P = 0.19) for tumors < or = 10 mm. The mean time required for resection was longer for ESD than for EMR ( P < 0.001). Perforation and bleeding requiring blood transfusion occurred in a small percentage in the ESD group, but in none in the EMR group. CONCLUSION: In this study, EMR was comparable to ESD for the millimeter-sized lesions. We suggest that such small lesions might be well suited to treatment with EMR. PMID- 19681025 TI - Comparative study of the hygienic behavior of Carniolan and Africanized honey bees directed towards grouped versus isolated dead brood cells. AB - In Apis mellifera, hygienic behavior involves recognition and removal of sick, damaged or dead brood from capped cells. We investigated whether bees react in the same way to grouped versus isolated damaged capped brood cells. Three colonies of wild-type Africanized honey bees and three colonies of Carniolan honey bees were used for this investigation. Capped worker brood cells aged 12 to 14 days old were perforated with the pin-killing method. After making holes in the brood cells, the combs were placed back into the hives; 24 h later the number of cleaned cells was recorded in areas with pin-killed and control brood cells. Four repetitions were made in each colony. Isolated cells were more frequently cleaned than grouped cells, though variance analysis showed no significant difference (P = 0.1421). Carniolan bees also were somewhat, though not significantly more hygienic than Africanized honey bees (P = 0.0840). We conclude that honey bees can detect and remove both isolated and grouped dead brood. The tendency towards greater hygienic efficiency directed towards grouped pin-killed brood may be a consequence of a greater concentration of volatiles emanating from the wounds in the dead pupae. PMID- 19681024 TI - Latissimus dorsi/rib intercostal perforator myo-osseocutaneous free flap reconstruction in composite defects of the scalp: case series and review of literature. AB - Adequate coverage of complex, composite scalp defects in previously radiated, infected, or otherwise compromised tissue represents a challenge in reconstructive surgery. To provide wound closure with bony protection to the brain, improve cranial contour, and prevent or seal cerebrospinal fluid (CSF) leaks, composite free tissue transfer is a reliable and safe option. We report our experience with the latissimus dorsi/rib intercostal perforator myo osseocutaneous free flap in the reconstruction of bony and soft tissue defects of the cranium and overlying scalp. The surgical technique, design, and outcomes of the latissimus dorsi/rib intercostal perforator myo-osseocutaneous free flap reconstruction in five patients with cranial defects between 2003 and 2007 were retrospectively evaluated. Patient characteristics, defect size, underlying cause, reconstructive details, and complications were analyzed. All patients (age 43 to 81) had composite defects ranging from 36 to 750 cm2 (mean size 230 cm2) for the bony component and from 16 to 400 cm2 (mean size 170 cm2) for the soft tissue defect. All patients had a history of prior or current infection of the affected area, and two patients had a CSF leak. Defects were due to malignancy and infection (n = 2), infiltrative cutaneous mucormycosis with osteomyelitis (n = 1), and hemorrhagic stroke requiring craniectomy (n = 2), complicated by infection and failed cranioplasty in one patient and continuous CSF leak in the other. The latissimus dorsi composite free flap consisting of skin, muscle, and vascularized rib can successfully cover large complex cranial defects, provide skeletal support, improve contour, and significantly enhance functional outcome with limited donor site morbidity. PMID- 19681026 TI - Pollination of tomatoes by the stingless bee Melipona quadrifasciata and the honey bee Apis mellifera (Hymenoptera, Apidae). AB - The pollination effectiveness of the stingless bee Melipona quadrifasciata and the honey bee Apis mellifera was tested in tomato plots. The experiment was conducted in four greenhouses as well as in an external open plot in Ribeirao Preto, SP, Brazil. The tomato plants were exposed to visits by M. quadrifasciata in one greenhouse and to A. mellifera in another; two greenhouses were maintained without bees (controls) and an open field plot was exposed to pollinators in an area where both honey bee and stingless bee colonies are abundant. We counted the number of tomatoes produced in each plot. Two hundred tomatoes from each plot were weighed, their vertical and transversal circumferences were measured, and the seeds were counted. We collected 253 Chrysomelidae, 17 Halictidae, one Paratrigona sp, and one honey bee from the flowers of the tomato plants in the open area. The largest number of fruits (1414 tomatoes), the heaviest and largest tomatoes, and the ones with the most seed were collected from the greenhouse with stingless bees. Fruits cultivated in the greenhouse with honey bees had the same weight and size as those produced in one of the control greenhouses. The stingless bee, M. quadrifasciata, was significantly more efficient than honey bees in pollinating greenhouse tomatoes. PMID- 19681028 TI - Bee communities (Hymenoptera: Anthophila) of the "Cerrado" ecosystem in Sao Paulo State, Brazil. AB - Five surveys of the bee communities in four "Cerrado" ecosystem reserves in Sao Paulo State were compared for species richness and similarity. These areas are fragment vegetation reser-ves located in the Cerrado Corumbatai Reserve (Corumbatai), Jatai Ecological Park (Luiz Antonio), Cajuru (Cajuru), and Vassununga State Park - "Gleba de Cerrado de Pe-de-Gigante" (Santa Rita do Passa Quatro). The methodology consisted of capturing bees foraging on flowers along transects, though with small differences between surveys. These "cerrado" areas have a large number of species of native bees, which are important pollinators in several Brazilian ecosystems. The community of bees varied among these different fragments. Based on 500 individuals (standardized by rarefaction), Cajuru, Corumbatai 1 and Corumbatai 2 were the areas with highest species richness, and Jatai and Pe-de-Gigante had the lowest species richness in the bee communities. The bee faunas of Corumbatai 2 and Pe-de-Gigante had the highest similarity, forming a group with the bee fauna of Cajuru. The bee faunas of Corumbatai 1 and Jatai were isolated from this group. We found that the bee species richness and similarity found in these "cerrado" areas cannot be explained by general factors such as the size of the fragment, the species richness of plants and the distance between the areas. Therefore, we suppose that local factors that differ among areas, such as interactions between populations, and competition and interference from surrounding areas influence and determine bee species richness and similarity in these reserves. PMID- 19681027 TI - Does beekeeping reduce genetic variability in Melipona scutellaris (Apidae, Meliponini)? AB - Many factors have contributed to reductions in wild populations of stingless bees, such as: deforestation, displacement and destruction of nests by honey gatherers, as well as use of insecticides and other agrochemicals. All of these can potentially affect the populational structure of native species. We analyzed genetic variability and populational structure of Melipona scutellaris, based on five microsatellite loci, using heterologous primers of M. bicolor. Samples were taken from 43 meliponaries distributed among 30 sites of four northeastern states of Brazil (Pernambuco, Alagoas, Sergipe, and Bahia). Thirty-one alleles were found to be well distributed among the populations, with sizes ranging from 85 to 146 bp. In general, there was a variable distribution and frequency of alleles among populations, with either exclusive and/or fixed alleles at some sites. The population of Pernambuco was the most polymorphic, followed by Bahia, Alagoas and Sergipe. The heterozygosity was Ho = 0.36 on average, much lower than what has been reported for M. bicolor (Ho = 0.65). Most populations were not under Hardy Weinberg equilibrium. We found a higher variation within rather than among populations, indicating no genetic structuring in those bees maintained in meliponaries. This apparent homogenization may be due to intense beekeeping activity, including exchange of genetic material among beekeepers. Based on our findings, we recommend more studies of meliponaries and of wild populations in order to help orient management and conservation of these native pollinators. PMID- 19681029 TI - Experimental studies of the distribution of gene frequencies in very small populations of Drosophila melanogaster: I. Forked. 1954. PMID- 19681030 TI - Variation in genotypic responses and biochemical analysis of callus induction in cultivated wheat. AB - Wheat is notorious for callus induction, which is a major hindrance in direct gene transfer and consequently for genetic improvement programs. In order to provide a successful platform for gene transformation, good callus quantity and quality is important. We investigated the variation in callus induction capabilities of Pakistani wheat cultivars and measured the reducing sugar content in the induced calluses. Ten elite wheat varieties, developed and cultivated in Pakistan were selected on the basis of agronomic and stress tolerance parameters. Significant differences were found between and among wheat cultivars for callus induction response, shoot length and callus quality. The callus induction responses of Punjab-81, Punjab-96 and Zarghoon-79 were found to be the best among the 10 varieties. The induced calluses were of two types, embryogenic (hard) and non-embryogenic (soft). The seeds gave good germination. The highest reducing sugar concentration was found in cultivar Sutlaj-86, which needs to be tested for stress resistance, a measure of its utility for genetic engineering programs. The relative callus induction rate and reducing sugar content of the wheat cultivars were found to be genotype-dependent. PMID- 19681031 TI - Frequency of the S65C mutation in the hemochromatosis gene in Brazil. AB - Development of hereditary hemochromatosis is associated with the C282Y, H63D or S65C mutations in the hemochromatosis gene. Though there is extensive knowledge about the former two, there is little information on the mechanism of action and the allelic frequency of the S65C mutation. We examined the prevalence of the S65C mutation of the hemochromatosis gene in Brazilians with clinical suspicion of hereditary hemochromatosis. Genotyping for this mutation was carried out in 633 individuals with clinical suspicion of hereditary hemochromatosis, using the polymerase chain reaction, followed by enzymatic digestion. The sample comprised 77.1% men and 22.9% women, giving a ratio of approximately 3:1; the mean age was 48.8 +/- 13.8 years. More than half (57.3%) of the individuals in the sample were 41 to 60 years old. The frequency of heterozygotes for this mutation was 0.016; no homozygous mutant patients were found. This is the first analysis of the S65C mutation in individuals suspected of having hereditary hemochromatosis in Brazil. PMID- 19681032 TI - Single nucleotide polymorphisms from Theobroma cacao expressed sequence tags associated with witches' broom disease in cacao. AB - In order to increase the efficiency of cacao tree resistance to witches' broom disease, which is caused by Moniliophthora perniciosa (Tricholomataceae), we looked for molecular markers that could help in the selection of resistant cacao genotypes. Among the different markers useful for developing marker-assisted selection, single nucleotide polymorphisms (SNPs) constitute the most common type of sequence difference between alleles and can be easily detected by in silico analysis from expressed sequence tag libraries. We report the first detection and analysis of SNPs from cacao-M. perniciosa interaction expressed sequence tags, using bioinformatics. Selection based on analysis of these SNPs should be useful for developing cacao varieties resistant to this devastating disease. PMID- 19681033 TI - Molecular characterization of wheat germplasm using microsatellite markers. AB - We investigated the genetic diversity of 63 wheat genotypes, composed of 48 accessions and 15 varieties, using 56 polymorphic simple sequence repeat primers. One hundred and eighty-six loci were found, with a mean of 131.26 alleles per locus. Cluster analysis based on microsatellite allelic diversity discriminated the accessions and varieties into different clusters; genetic diversity was the highest between variety Kohistan-97 and accession number 011512, giving a genetic similarity value of 0.4198. Accession numbers 011484 and 011356 gave a genetic similarity value of 0.9589, indicating that these accessions were 95.89% similar. We found that microsatellite markers could characterize and discriminate all of the genotypes; more primers could be used for saturation of different regions in further studies. PMID- 19681034 TI - Abnormal glucose tolerance is associated with diminished postload change in leptin levels in women. AB - BACKGROUND: It is generally accepted that the metabolic effects of leptin are diminished in the obese due to leptin resistance. Hormone resistance may develop if diurnal (including meal-related) changes in hormone levels are disrupted. We sought to describe leptin changes after a 75 g oral glucose tolerance test (OGTT) in women with a prior diagnosis of gestational diabetes mellitus (a high risk group for the metabolic syndrome) compared to that in healthy controls. METHODS: In 2000 a retrospective cohort study was performed on women who had been diagnosed with gestational diabetes mellitus (WHO criteria 1985, n = 57) between 1996 and 1998 and on a healthy control female group (n = 36) all of whom had had a prior pregnancy without any diagnosis of diabetes. All the women underwent a standard 75 g OGTT. Serum leptin was measured by radioimmunoassay before and 90 min after the OGTT. RESULTS: Using multilevel models of change, fasting leptin levels were shown to be associated with body mass index; 10.1% (95% CI 8.1-12.1%) increase per 1 kg/m(2) increase in body mass index), homeostasis model assessment insulin sensitivity; 0.4% (95% CI 0.2-0.7%) decrease per 1% increase in insulin sensitivity); abnormal glucose tolerance (24% decrease, 95% CI 8-37%); and smoking (31% decrease, 95% CI 16-44%). Postload (90 min) leptin levels decreased significantly in women with normal glucose tolerance by 13% (95% CI 8-18%), while no significant change in postload leptin level was apparent in women with abnormal glucose tolerance (3% increase, 95% CI -4% to 29%). CONCLUSIONS: Disturbed leptin changes were found following an OGTT in women with abnormal glucose tolerance that might be either a cause or a consequence of leptin resistance. PMID- 19681035 TI - Microvascular dysfunction in diabetic foot disease and ulceration. AB - Diabetic foot disease and ulceration is a major complication that may lead to the amputation of the lower limbs. Microangiopathy may play a significant role in the pathogenesis of tissue breakdown in the diabetic foot. However, the precise mechanisms of this process remain unclear and poorly understood. Microvasculature in the skin is comprised of nutritive capillaries and thermoregulatory arteriovenous shunt flow. It is regulated through the complex interaction of neurogenic and neurovascular control. The interplay among endothelial dysfunction, impaired nerve axon reflex activities, and microvascular regulation in the diabetic patient results in the poor healing of wounds. Skin microvasculature undergoes both morphologic changes as well as functional deficits when parts of the body come under stress or injury. Two important theories that have been put forward to explain the abnormalities that have been observed are the haemodynamic hypothesis and capillary steal syndrome. With advances in medical technology, microvasculature can now be measured quantitatively. This article reviews the development of microvascular dysfunction in the diabetic foot and discusses how it may relate to the pathogenesis of diabetic foot problems and ulceration. Common methods for measuring skin microcirculation are also discussed. PMID- 19681036 TI - Immobilized protein-polymer nanoreactors. PMID- 19681037 TI - Resonance Raman spectroscopy as an effective tool for the determination of antioxidative stability of cosmetic formulations. AB - Carotenoids beta-carotene, lutein, lycopene and others are well-known powerful antioxidants acting as an effective neutralizer of free radicals produced in the human organism as a result of the influence of stress factors, such as UV irradiation. The protective effect of antioxidants is used in cosmetic products to increase the skin protection against the destructive action of free radicals and for the stabilization of formulations against oxidation. In the skin, the different antioxidant substances form protection chains to avoid their destruction by the interaction with the free radicals. Similar effects have to be expected also in topically applied formulations. In the present study the influence of different mixtures of antioxidants (beta-carotene, vitamins C and E) on the stability of antioxidants in formulations used for skin treatment was investigated. The measurements were carried out by using non-invasive resonance Raman spectroscopy for the detection of the carotenoid concentration in the cosmetic formulations. PMID- 19681038 TI - Slug is a downstream mediator of transforming growth factor-beta1-induced matrix metalloproteinase-9 expression and invasion of oral cancer cells. AB - Members of Snail family of transcription factors play an important role in oral cancer progression by inducing epithelial-mesenchymal transition, by promoting invasion and by increasing matrix metalloproteinase (MMP) expression. Although Snail (Snai1) is the best characterized and the most extensively studied member of this family, the role and regulation of Slug (Snai2) in oral cancer progression is less well understood. In this report, we show that transforming growth factor-beta1 (TGF-beta1) increases Slug levels in tert-immortalized oral keratinocytes and in malignant oral squamous cell carcinoma (OSCC) cells. Inhibiting ERK1/2 signaling, but not PI3-kinase signaling, blocked TGF-beta1 induced Slug expression in the malignant UMSCC1 cells. To further examine the role of Slug in OSCC progression, we generated UMSCC1 cells with inducible expression of Slug protein. Induction of Slug in UMSCC1 cells did not repress E cadherin levels or regulate individual movement of UMSCC1 cells. Instead, Slug enhanced cohort migration and Matrigel invasion by UMSCC1 cells. Slug increased MMP-9 levels and MMP-9-specific siRNA blocked Slug-induced Matrigel invasion. Interestingly, Slug-specific siRNA attenuated TGF-beta1-induced MMP-9 expression and Matrigel invasion. These data demonstrate that TGF-beta1 increases Slug via ERK1/2 signaling, and thereby contributes to OSCC progression. PMID- 19681039 TI - Phosphorylation of phospholipase C-delta 1 regulates its enzymatic activity. AB - Phosphorylation of phospholipase C-delta(1) (PLC-delta(1)) in vitro and in vivo was investigated. Of the serine/threonine kinases tested, protein kinase C (PKC) phosphorylated the serine residue(s) of bacterially expressed PLC-delta(1) most potently. It was also demonstrated that PLC-delta(1) directly bound PKC-alpha via its pleckstrin homology (PH) domain. Using deletion mutants of PLC-delta(1) and synthetic peptides, Ser35 in the PH domain was defined as the PKC mediated in vitro phosphorylation site of PLC-delta(1). In vitro phosphorylation of PLC delta(1) by PKC stimulated [(3)H]PtdIns(4,5)P(2) hydrolyzing activity and [(3)H]Ins(1,4,5)P(3)-binding of the PLC-delta(1). On the other hand, endogenous PLC-delta(1) was constitutively phosphorylated and phosphoamino acid analysis revealed that major phosphorylation sites were threonine residues in quiescent cells. The phosphorylation level and the species of phosphoamino acid were not changed by various stimuli such as PMA, EGF, NGF, and forskolin. Using matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry, we determined that Thr209 of PLC-delta(1) is one of the constitutively phosphorylated sites in quiescent cells. The PLC activity was potentiated when constitutively phosphorylated PLC-delta(1) was dephosphorylated by endogenous phosphatase(s) in vitro. Additionally, coexpression with PKC-alpha reduced serine phosphorylation of PLC-delta(1) detected by an anti-phosphoserine antibody and PLC-delta(1)-dependent basal production of inositol phosphates in NIH-3T3 cells, suggesting PKC-alpha activates phosphatase or inactivates another kinase involved in PLC-delta(1) serine phosphorylation to modulate the PLC delta(1) activity in vivo. Taken together, these results suggest that PLC delta(1) has multiple phosphorylation sites and phosphorylation status of PLC delta(1) regulates its activity positively or negatively depends on the phosphorylation sites. PMID- 19681040 TI - Three-dimensional porous scaffolds at the crossroads of tissue engineering and cell-based gene therapy. AB - In the last 20 years, more than 1,500 gene therapy clinical trials have been approved worldwide targeting a variety of indications, from inherited monogenic diseases to acquired conditions such as cancer, cardiovascular and infectious diseases. However, concerns about the safety and efficacy of gene therapy pharmaceuticals justify the development of alternative strategies to ensure the clinical translation of this still promising field. In particular, ex vivo gene therapy strategies using autologous adult stem cells coupled to three-dimensional (3D) porous scaffolds show great promises in preclinical studies. Developments in the fields of biomaterial sciences and tissue engineering have already helped understanding how we can harness to regenerative potential of many cell types to create artificial tissues and organs and vastly improve the engraftment of ex vivo manipulated adult stem cells. In this article, we will review the current state of the art in tissue engineering by exploring the various types of clinically available biomaterials and the methods used to process them into complex 3D scaffolds. We will then review how these technologies are applied in cell-based gene therapy and identify novel avenues of research that may benefit patients in the near future. PMID- 19681041 TI - Productive infection of Piscirickettsia salmonis in macrophages and monocyte-like cells from rainbow trout, a possible survival strategy. AB - Piscirickettsia salmonis is the etiologic agent of the salmonid rickettsial septicemia (SRS), an endemic disease which causes significant losses in salmon production. This intracellular bacterium is normally cultured in salmonid epithelial cell lines inducing characteristic cytopathic effects (CPEs). In this study we demonstrate that P. salmonis is able to infect, survive, replicate, and propagate in the macrophages/monocytes cell line RTS11 derived from rainbow trout spleen, without inducing the characteristic CPEs and the host cells showing the same expression levels as non-infected control cell. On the other hand, bacteria were capable of expressing specific proteins within infected cells. Infected macrophages cease proliferation and a fraction of them detached from the plate, transform to non-adhesive, monocyte-like cells with proliferative activity. Productive infection of P. salmonis into salmonid macrophage/monocyte cells in culture provides an excellent model for the study of host-pathogen interactions, almost unknown in the case of P. salmonis. Our results suggest that the infection of cells from the salmonid innate immune system without inducing an important cell death response should lead to the persistence of the bacteria and consequently their dissemination to other tissues, favoring the evasion of the first line of defense against pathogens. PMID- 19681042 TI - Phosphate regulates embryonic endochondral bone development. AB - Phosphate is required for terminal differentiation of hypertrophic chondrocytes during postnatal growth plate maturation. In vitro models of chondrocyte differentiation demonstrate that 7 mM phosphate, a concentration analogous to that of the late gestational fetus, activates the mitochondrial apoptotic pathway in hypertrophic chondrocytes. This raises the question as to whether extracellular phosphate modulates chondrocyte differentiation and apoptosis during embryonic endochondral bone formation. To address this question, we performed investigations in the mouse metatarsal culture model that recapitulates in vivo bone development. Metatarsals were cultured for 4, 8, and 12 days with 1.25 and 7 mM phosphate. Metatarsals cultured with 7 mM phosphate showed a decrease in proliferation compared to those cultured in 1.25 mM phosphate. This decrease in proliferation was accompanied by an early enhancement in hypertrophic chondrocyte differentiation, associated with an increase in FGF18 expression. By 8 days in culture, an increase caspase-9 activation and apoptosis of hypertrophic chondrocytes was observed in the metatarsals cultured in 7 mM phosphate. Immunohistochemical analyses of embryonic bones demonstrated activation of caspase-9 in hypertrophic chondrocytes, associated with vascular invasion. Thus, these investigations demonstrate that phosphate promotes chondrocyte differentiation during embryonic development and implicate a physiological role for phosphate activation of the mitochondrial apoptotic pathway during embryonic endochondral bone formation. PMID- 19681043 TI - The biological activity of ubiquitinated BoNT/B light chain in vitro and in human SHSY-5Y neuronal cells. AB - BoNT/B light chain is a zinc-dependent endopeptidase. After entering its target, the neuronal cell, BoNT/B is responsible for synaptobrevin-2 (VAMP-2) cleavage. This results in reduced neurotransmitter (acetylcholine) release from synaptic vesicles, yielding muscular paralysis. Since the toxin persists in neuronal cells for an extended period, regeneration of VAMP-2 is prevented. We evaluated therapeutic targets to overcome botulinum persistence because early removal would rescue the neuronal cell. The ubiquitination/proteasome cellular pathway is responsible for removing "old" or undesirable proteins. Therefore, we assessed ubiquitination of BoNT/B light chain in vitro, and characterized the effects of ubiquitination modulating drugs, PMA (phorbol 12-myristate 13-acetate) and expoxomicin, on ubiquitination of BoNT/B light chain in neuronal cells. Both drugs altered BoNT/B light chain ubiquitination. Ubiquitination in vitro and in cells decreased the biological activity of BoNT/B light chain. These results further elucidate BoNT protein degradation pathways in intoxicated neuronal cells and mechanisms to enhance toxin removal. PMID- 19681044 TI - Microtubule-associated protein tau in human prostate cancer cells: isoforms, phosphorylation, and interactions. AB - Tau is a microtubule-associated protein whose function has been investigated primarily in neurons. Recently, tau expression has been correlated with increased drug resistance in various cancers of non-neuronal tissues. In this report, we investigate the tau expressed in cancerous prostate lines ALVA-31, DU 145, and PC 3. Prostate cancer tau is heat-stable and highly phosphorylated, containing many of the modifications identified in Alzheimer's disease brain tau. RT-PCR and phosphatase treatment indicated that all six alternatively spliced adult brain tau isoforms are expressed in ALVA-31 cells, and isoforms containing exon 6 as well as high molecular weight tau isoforms containing either exon 4A or a larger splice variant of exon 4A are also present. Consistent with its hyperphosphorylated state, a large proportion of ALVA-31 tau does not bind to microtubules, as detected by confocal microscopy and biochemical tests. Finally, endogenous ALVA-31 tau can interact with the p85 subunit of phosphatidylinositol 3-kinase, as demonstrated by co-immunoprecipitations and in vitro protein-binding assays. Our results suggest that tau in prostate cancer cells does not resemble that from normal adult brain and support the hypothesis that tau is a multifunctional protein. PMID- 19681045 TI - Caffeic acid phenethyl ester, an active component of honeybee propolis attenuates osteoclastogenesis and bone resorption via the suppression of RANKL-induced NF kappaB and NFAT activity. AB - Receptor activator NF-kappaB ligand (RANKL)-activated signaling is essential for osteoclast differentiation, activation and survival. Caffeic acid phenethyl ester (CAPE), a natural NF-kappaB inhibitor from honeybee propolis has been shown to have anti-tumor and anti-inflammatory properties. In this study, we investigated the effect of CAPE on the regulation of RANKL-induced osteoclastogenesis, bone resorption and signaling pathways. Low concentrations of CAPE (<1 microM) dose dependently inhibited RANKL-induced osteoclastogenesis in RAW264.7 cell and bone marrow macrophage (BMM) cultures, as well as decreasing the capacity of human osteoclasts to resorb bone. CAPE inhibited both constitutive and RANKL-induced NF kappaB and NFAT activation, concomitant with delayed IkappaBalpha degradation and inhibition of p65 nuclear translocation. At higher concentrations, CAPE induced apoptosis and caspase 3 activities of RAW264.7 and disrupts the microtubule network in osteoclast like (OCL) cells. Taken together, our findings demonstrate that inhibition of NF-kappaB and NFAT activation by CAPE results in the attenuation of osteoclastogenesis and bone resorption, implying that CAPE is a potential treatment for osteolytic bone diseases. PMID- 19681046 TI - Matrix association of latent TGF-beta binding protein-2 (LTBP-2) is dependent on fibrillin-1. AB - The components of the extracellular matrix (ECM) and their differential expression patterns play important roles in tissue formation. The deposition of latent TGF-beta binding proteins (LTBPs) to the ECM exhibit distinct distribution profiles. We have analyzed here the temporal and spatial ECM association of latent TGF-beta binding protein LTBP-2 in cultured human embryonic lung fibroblasts. We found that LTBP-2 was not assembled to the ECM until by confluency of cultures following the deposition of fibronectin (FN) and fibrillin 1. In 5-day-old cultures LTBP-2 was rapidly secreted from cells and it subsequently associated with the ECM as shown by metabolic labeling and immunoprecipitation. LTBP-2 colocalized transiently with fibronectin and failed to assemble to the ECM of FN deficient mouse fibroblasts. Analysis of different cultured human cell lines revealed partial colocalization of LTBP-2 and fibrillin 1 in the ECM of fibroblasts, MG-63 osteosarcoma cells and human vascular endothelial cells. Silencing of fibrillin-1 expression by lentiviral shRNAs profoundly disrupted the deposition of LTBP-2. Current results suggest that LTBP 2 is not an element of the provisional ECM of fibroblasts but is more likely a component of more mature ECM and indicate that matrix association of LTBP-2 depends on a pre-formed fibrillin-1 network. PMID- 19681047 TI - Kinetics of transport and phosphorylation of glucose in cancer cells. AB - Metabolic control analysis of tumor glycolysis has indicated that hexokinase (HK) and glucose transporter (GLUT) exert the main flux control (71%). To understand why they are the main controlling steps, the GLUT and HK kinetics and the contents of GLUT1, GLUT2, GLUT3, GLUT4, HKI, and HKII were analyzed in rat hepatocarcinoma AS-30D and HeLa human cervix cancer. An improved protocol to determine the kinetic parameters of GLUT was developed with D-[2-(3)H-glucose] as physiological substrate. Kinetic analysis revealed two components at low- and high-glucose concentrations in both tumor cells. At low glucose and 37 degrees C, the V(max) was 55 +/- 20 and 17.2 +/- 6 nmol (min x mg protein)(-1), whereas the K(m) was 0.52 +/- 0.7 and 9.3 +/- 3 mM for hepatoma and HeLa cells, respectively. GLUT activity was partially inhibited by cytochalasin B (IC(50) = 0.44 +/- 0.1; K(i) = 0.3 +/- 0.1 microM) and phloretin (IC(50) = 8.7 microM) in AS-30D hepatocarcinoma. At physiological glucose, GLUT1 and GLUT3 were the predominant active isoforms in HeLa cells and AS-30D cells, respectively. HK activity in HeLa cells was much lower (60 mU/mg protein) than that in AS-30D cells (700 mU/mg protein), but both HKs were strongly inhibited by G6P. HKII was the predominant isoform in AS-30D carcinoma and HeLa cells. The much lower GLUT V(max) and catalytic efficiency (V(max)/K(m)) values in comparison to those of G6P-sensitive HK suggested the transporter exerts higher control on the glycolytic flux than HK in cancer cells. Thus, GLUT seems a more adequate therapeutic target. PMID- 19681048 TI - NACE-ESI-TOF MS to reveal phenolic compounds from olive oil: introducing enriched olive oil directly inside capillary. AB - Most CE methods for the analysis of phenols from olive oil use an aqueous electrolyte separation medium, although the importance of NACE is obvious, as this kind of CE seems to be more compatible with the hydrophobic olive oil matrix and could facilitate its direct injection. In the current work we develop a method involving SPE and NACE coupled to ESI-TOF MS. All the CE and ESI-TOF MS parameters were optimized in order to maximize the number of phenolic compounds detected and the sensitivity in their determination. Electrophoretic separation was carried out using a CE buffer system consisting of 25 mM NH(4)OAc/AcH in methanol/ACN (1/1 v/v) at an apparent pH value of 5.0. We studied in depth the effect of the nature and concentration of different electrolytes dissolved in different organic solvents and other experimental and instrumental CE variables. The results were compared with those obtained by CZE (with aqueous buffers) coupled to ESI-TOF MS; both methods offered to the analyst the chance to study phenolic compounds of different families (such as phenolic alcohols, lignans, complex phenols, flavonoids, etc.) from virgin olive oil by injecting methanolic extracts with efficient and fast CE separations. In the case of NACE method, we also studied the direct injection of the investigated matrix introducing a plug of olive oil directly into the capillary. PMID- 19681049 TI - CE separation approaches for combinations of anthracyclines and taxanes. AB - For the first time the versatility of CE is demonstrated for the separation of different types of anticancer drugs - anthracyclines and taxanes simultaneously. The use of these drugs in combination therapy for cancer has sparked interest in the development of methods for potential application. The simultaneous analysis of anthracyclines and taxanes can significantly increase a sample throughput of a clinical laboratory. The study shows the potential of CE for such a challenge: anthracyclines and taxanes were separated by CZE, MEKC and MEEKC. The MEEKC method was successfully applied to these compounds for the first time and was characterised by very short separation time, high efficiencies of peaks and was proven to be generic for the separation of different combinations of anthracyclines and taxanes. This separation approach could be highly beneficial for clinical analysis if applied with a sensitive detection system. MEKC and high speed MEEKC methods were proven to show good potential in their application to plasma samples. PMID- 19681050 TI - Expanding the scope of CE reactor to ssDNA-binding protein-ssDNA complexes as exemplified for a tool for direct measurement of dissociation kinetics of biomolecular complexes. AB - CE reactor (CER), which was developed as a tool for direct measurement of the dissociation kinetics of metal complexes, was successfully applied to the complexes of Escherichia coli ssDNA-binding protein (SSB) with ssDNA. The basic concept of CER is the application of CE separation process as a dissociation kinetic reactor for the complex, and the observation of the on-capillary dissociation reaction profile of the complex as the decrease of the peak height of the complex with increase of the migration time. The peak height of [SSB ssDNA] decreases as the migration time increases since the degree of the decrease of [SSB-ssDNA] through the on-capillary dissociation reaction is proportional to the degree of the decrease of the peak height of [SSB-ssDNA]. The dissociation degree-time profiles for the complexes are quantitatively described by analyzing a set of electropherograms with different migration times. Dissociation rate constants of [SSB-ssDNA] consisting of 20-mer, 25-mer and 31-mer ssDNA were directly determined to be 3.99x10(-4), 4.82x10(-4) and 1.50x10(-3)/s, respectively. CER is a concise and effective tool for dissociation kinetic analysis of biomolecular complexes. PMID- 19681051 TI - Determination of enantiomerization barrier of thioridazine by dynamic capillary electrophoresis using sulfated cyclodextrins as chiral selectors. AB - The enantiomerization of thioridazine (THD) using sulfated beta-CDs (S-beta-CDs) as chiral selectors in a citrate buffer at pH 3.0 was investigated by dynamic CE. The enantiomers of THD were well separated with dual CD systems consisting of S beta-CD and a neutral CD. The electropherograms featuring a plateau formation, which indicated the occurrence of the enantiomerization of THD were obtained. The unified equation implemented in the software program DCXplorer was employed to evaluate elution profiles and to determine rate constants of the enantiomerization of THD. Activation parameters were evaluated from temperature dependent measurements between 15 and 25 degrees C with an increment of 2 degrees C. The enantiomerization barriers of THD in two different electrophoretic systems were determined. Comparative studies on enantioseparation of THD using S-beta-CDs with different degree of substitution and positions of sulfate substituent, such as randomly sulfate-substituted beta-CD, 18-sulfate-substituted beta-CD and heptakis(2,3-dihydroxy-6-O-sulfo)-beta-CD reveal that the interactions between chiral selectors and THD plays an important role in the enantioseparation and enantiomerization of THD. PMID- 19681052 TI - Development of micropump-actuated negative pressure pinched injection for parallel electrophoresis on array microfluidic chip. AB - A micropump-actuated negative pressure pinched injection method is developed for parallel electrophoresis on a multi-channel LIF detection system. The system has a home-made device that could individually control 16-port solenoid valves and a high-voltage power supply. The laser beam is excitated and distributes to the array separation channels for detection. The hybrid Glass-PDMS microfluidic chip comprises two common reservoirs, four separation channels coupled to their respective pneumatic micropumps and two reference channels. Due to use of pressure as a driving force, the proposed method has no sample bias effect for separation. There is only one high-voltage supply needed for separation without relying on the number of channels, which is significant for high-throughput analysis, and the time for sample loading is shortened to 1 s. In addition, the integrated micropumps can provide the versatile interface for coupling with other function units to satisfy the complicated demands. The performance is verified by separation of DNA marker and Hepatitis B virus DNA samples. And this method is also expected to show the potential throughput for the DNA analysis in the field of disease diagnosis. PMID- 19681053 TI - Contribution of the fibrinolytic pathway to hematopoietic regeneration. AB - Hematopoietic stem cells (HSCs) can differentiate and proliferate in response to hematopoietic stress (e.g., myelosuppression, infections, and allergic reactions), thereby ensuring a well-regulated supply of mature and immature hematopoietic cells within the circulation and prompt adjustment of blood cell levels within normal ranges. The recovery of tissues and organs from hematopoietic stress (e.g., myelosuppression or ionizing irradiation) is dependent on two cell types: resident HSCs which repopulate the bone marrow (BM) cavity, and stromal cells. BM regeneration critically depends on the release of soluble factors from cells such as stromal cells, a process regulated by proteases. Two proteolytic systems, the fibrinolytic system and the matrix metalloproteinases (MMPs), have recently been shown to be involved in this process (Heissig B, 2007, Cell Stem Cell 1: 658-670). The plasminogen/plasmin system is mostly recognized for its fibrinolytic activity, but it is also involved in processes such as cell invasion, chemotaxis, growth factor activity modulation, and tissue remodeling. This review focuses on the role of plasmin and its activators as key players in controlling the hematopoietic stress response after myelosuppression (hematopoietic regeneration). Aspects of plasmin regulation, especially regulation of its ability to activate MMPs and the functional consequences of this enzyme activation, such as plasmin-mediated release of biologically relevant cytokines from the matrix and cell surfaces, will be discussed. PMID- 19681054 TI - Biomarkers of lung-related diseases: current knowledge by proteomic approaches. AB - The lung epithelial surface is one of the vital barriers or sensors in the body responding to the external atmosphere and thereby always subjecting to direct toxicological exposure, stress, stimulus, or infection. Due to its relatively higher sensitivity in response to toxicants, the use of lung epithelial cell culture and lung tissue from animal models or patients has facilitated our learning to lung physiopathology and toxicopharmacology. The recent advancement of proteomics has made it possible to investigate the cellular response at a global level. In this review, the potential applications of proteomic approach in studying lung-related diseases and biomarker discovery will be discussed. PMID- 19681055 TI - A novel peak detection approach with chemical noise removal using short-time FFT for prOTOF MS data. AB - Peak detection is a pivotal first step in biomarker discovery from MS data and can significantly influence the results of downstream data analysis steps. We developed a novel automatic peak detection method for prOTOF MS data, which does not require a priori knowledge of protein masses. Random noise is removed by an undecimated wavelet transform and chemical noise is attenuated by an adaptive short-time discrete Fourier transform. Isotopic peaks corresponding to a single protein are combined by extracting an envelope over them. Depending on the S/N, the desired peaks in each individual spectrum are detected and those with the highest intensity among their peak clusters are recorded. The common peaks among all the spectra are identified by choosing an appropriate cut-off threshold in the complete linkage hierarchical clustering. To remove the 1 Da shifting of the peaks, the peak corresponding to the same protein is determined as the detected peak with the largest number among its neighborhood. We validated this method using a data set of serial peptide and protein calibration standards. Compared with MoverZ program, our new method detects more peaks and significantly enhances S/N of the peak after the chemical noise removal. We then successfully applied this method to a data set from prOTOF MS spectra of albumin and albumin-bound proteins from serum samples of 59 patients with carotid artery disease compared to vascular disease-free patients to detect peaks with S/N> or =2. Our method is easily implemented and is highly effective to define peaks that will be used for disease classification or to highlight potential biomarkers. PMID- 19681056 TI - Soluble TNF-alpha but not transmembrane TNF-alpha sensitizes T cells for enhanced activation-induced cell death. AB - In addition to its proinflammatory effects, TNF-alpha exhibits immunosuppression. Here, we compared the capacities of transmembrane TNF-alpha (tmTNF) and soluble TNF-alpha (sTNF) in regulating expansion of activated T cells by apoptosis. Splenic CD4(+) T cells from wtTNF, TNF-alpha-deficient (TNF(-/-)) and TNF(-/-) mice expressing a non-cleavable mutant tmTNF showed comparable proliferation rates upon TCR-mediated stimulation. Activation-induced cell death (AICD), however, was significantly attenuated in tmTNF and TNF(-/-), compared with wtTNF CD4(+) T cells. Addition of sTNF during initial priming was sufficient to enhance susceptibility to AICD in tmTNF and TNF(-/-) CD4(+) T cells to levels seen in wtTNF CD4(+) T cells, whereas addition of sTNF only during restimulation failed to enhance AICD. sTNF-induced, enhanced susceptibility to AICD was dependent on both TNF receptors. The reduced susceptibility of tmTNF CD4(+) T cells for AICD was also evident in an in vivo model of adoptively transferred CD4(+) T-cell mediated colonic inflammation. Hence, the presence of sTNF during T-cell priming may represent an important mechanism to sensitize activated T cells for apoptosis, thereby attenuating the extent and duration of T-cell reactivities and subsequent T-cell-mediated, excessive inflammation. PMID- 19681057 TI - Shifting therapeutic attention in MS to osteopontin, type 1 and type 2 IFN. AB - It is widely debated whether MS is mediated solely by Th1, or solely by Th17, whether it might be mediated by both pathways, or perhaps by neither pathway. We must integrate the following four facts: first, MS lesions have a signature of IL 6-, IL-17-, osteopontin- and IFN-driven transcriptional activity. Second, MS is worsened with administration of IFN-gamma, the quintessential type 2 IFN and Th1 cytokine. Third, blockade of TNF-alpha worsened MS in clinical trials. Fourth, inhibiting the main driver of Th17, IL-23, failed to modulate relapsing-remitting MS (RRMS). Against this backdrop, standing outside the framework of the Th1 and Th17 pathways are the type 1 IFN, notably IFN-beta, the most widely used approved therapy for RRMS. A paper in this issue of the European Journal of Immunology demonstrates that IFN-beta suppresses production in CD4(+) T cells of both osteopontin and IL-17. In this commentary, the roles of these two molecules, i.e. osteopontin and IL-17, are discussed in relation to the pathogenesis of MS. Osteopontin may be more important than Th1 or Th17 in the pathogenesis of RRMS. Trials targeting this small integrin-binding protein ought to be pursued in RRMS. PMID- 19681058 TI - Acceleration of germination and early growth of wheat and bean seedlings grown under various magnetic field and osmotic conditions. AB - Magnetic field (MF) can have different effects on plant metabolism depending on its application style, intensity, and environmental conditions. This study reports the effects of different intensities of static MF (4 or 7 mT) on seed germination and seedling growth of bean or wheat seeds in different media having 0, 2, 6, and 10 atmosphere (atm) osmotic pressure prepared with sucrose or salt. The germination percentages of the treated seeds were compared with untreated seeds germinated in different osmotic pressure during 7 days of incubation. The application of both MFs promoted the germination ratios of bean and wheat seeds, regardless of increasing osmotic pressure of sucrose or salt. Growth data measured on the 7th day showed that the treated plants grew faster than control. After 7 days of incubation, the mean length of treated seedlings was statistically higher than control plants in all the media. The greatest germination and growth rates in both plants were from the test groups exposed to 7 mT MF. Strikingly, effects of static MF on germination and growth increased positively with increasing osmotic pressure or salt stress compared to their respective controls. On the other hand, MF application caused an increase in dry biomass accumulation of root and shoots of both seedlings; however, this effect was found statistically important in all the conditions for wheat but not for bean, in general. PMID- 19681059 TI - Idiopathic environmental intolerance attributed to electromagnetic fields (formerly 'electromagnetic hypersensitivity'): An updated systematic review of provocation studies. AB - Idiopathic Environmental Intolerance attributed to electromagnetic fields (IEI EMF; formerly 'electromagetic hypersensitivity') is a medically unexplained illness in which subjective symptoms are reported following exposure to electrical devices. In an earlier systematic review, we reported data from 31 blind provocation studies which had exposed IEI-EMF volunteers to active or sham electromagnetic fields and assessed whether volunteers could detect these fields or whether they reported worse symptoms when exposed to them. In this article, we report an update to that review. An extensive literature search identified 15 new experiments. Including studies reported in our earlier review, 46 blind or double blind provocation studies in all, involving 1175 IEI-EMF volunteers, have tested whether exposure to electromagnetic fields is responsible for triggering symptoms in IEI-EMF. No robust evidence could be found to support this theory. However, the studies included in the review did support the role of the nocebo effect in triggering acute symptoms in IEI-EMF sufferers. Despite the conviction of IEI-EMF sufferers that their symptoms are triggered by exposure to electromagnetic fields, repeated experiments have been unable to replicate this phenomenon under controlled conditions. A narrow focus by clinicians or policy makers on bioelectromagnetic mechanisms is therefore, unlikely to help IEI-EMF patients in the long-term. PMID- 19681060 TI - N-acetylcysteine improves renal hemodynamics in rats with cisplatin-induced nephrotoxicity. AB - This work investigated the effect of N-acetylcysteine (NAC), on renal hemodynamics in cisplatin (CP)-induced nephrotoxicity in Wistar-Kyoto (WKY) rats. The animals were divided into four groups (n = 5 or 6). The first and second groups received normal saline (control) and intraperitoneal (i.p.) N acetylcysteine (500 mg kg(-1) per day for 9 days), respectively. The third and fourth groups were given a single intraperitoneal (i.p.) injection of CP (5 mg kg(-1)) and an i.p. injection of CP (5 mg kg(-1)) together with i.p. NAC (500 mg kg(-1) per day for 9 days), respectively. At the end of the experiment, rats were anesthetized and blood pressure and renal blood flow were monitored, followed by intravenous (i.v.) injection of norepinephrine (NE) for measurement of renal vasoconstrictor responses. CP caused a significant reduction in renal blood flow but did not affect NE-induced renal vasoconstriction. In addition, CP significantly increased plasma concentrations of urea and creatinine and urinary N-acetyl-beta-D-glucosaminidase (NAG) activity and kidney relative weight. CP decreased body weight and creatinine clearance. Histopathologically, CP caused remarkable renal damage compared with control. NAC alone did not produce any significant change in any of the variables measured. However, NAC significantly ameliorated CP-induced hemodynamic, biochemical and histopathological changes. The concentration of platinum in the kidneys of CP ? NAC treated rats was less than in CP-treated rats by 37%. The results show that administration of i.p. NAC (500 mg kg(-1) per day for 9 days) reversed the renal hemodynamic changes as well as the biochemical and histopathological indices of CP-induced nephrotoxicity in WKY rats. PMID- 19681061 TI - The role of children's routines of daily living, supervision, and maternal fatigue in preschool children's injury risk. AB - The purpose of this study was to (a) explore the relationship between the lack of everyday routines and children's unintentional injuries, (b) examine two mechanisms of effect through which lack of routines might operate, children's sleep and maternal supervision, and (c) explore the influence of maternal fatigue in routines and maternal supervision. The sample comprised 264 mothers and their 3-year-old children. Routines were not directly related to children's injuries, but they moderated the relationship between maternal supervision and injuries. Lack of supervision was positively related to injuries only for children with infrequent everyday routines. Infrequent everyday routines were also related to inadequate children's sleep and maternal fatigue was inversely related to routines and maternal supervision. PMID- 19681062 TI - Further evidence that the rs1858830 C variant in the promoter region of the MET gene is associated with autistic disorder. AB - Previous studies in three independent cohorts have shown that the rs1858830 C allele variant in the promoter region of the MET gene on chromosome 7q31 is associated with autism. Another study has found correlations between other alterations in the MET gene and autism in two unrelated cohorts. This study screened two cohorts, an Autistic Disorder cohort from South Carolina and a Pervasive Developmental Disorder (PDD) cohort from Italy, for the presence of the C allele variant in rs1858830. A significant increase in the C allele variant frequency was found in the South Carolina Autistic Disorder patients as compared to South Carolina Controls (chi(2)=5.8, df=1, P=0.02). In the South Carolina cohort, a significant association with Autistic Disorder was found when comparing the CC and CG genotypes to the GG genotype (odds ratio (OR)=1.64; 95% confidence interval (CI)=1.12-2.40; chi(2)=6.5, df=1, P=0.01) in cases and controls. In the Italian cohort, no significant association with PDD was found when comparing the CC or CG genotype to the GG genotype (OR=1.20; 95% CI=0.56-2.56; chi(2)=0.2, df=1, P=0.64). This study is the third independent study to find the rs1858830 C variant in the MET gene promoter to be associated with autism. PMID- 19681063 TI - Rituximab (B-cell depleting antibody) associated lung injury (RALI): a pediatric case and systematic review of the literature. AB - INTRODUCTION: Pulmonary toxicity of delayed onset is a rare complication of B lymphocyte depleting antibody therapy and has been almost exclusively reported in older patients with B-cell malignancies. AIMS: To describe a pediatric patient with rituximab-associated lung injury (RALI), to systematically analyze previous reports of pulmonary complications, and to summarize common clinico-pathological features, treatment, and outcome. RESULTS: A teenage boy with focal segmental glomerulosclerosis (FSGS) presented with progressive dyspnea, fever, hypoxemia and fatigue 18 days after the completion of a second course of rituximab infusions for calcineurin inhibitor-dependent nephrotic syndrome. Respiratory symptoms started while he received high-dose prednisone for persistent proteinuria. Bilateral, diffuse ground-glass infiltrates corresponded to the presence of inflammatory cells in the bronchioalveolar lavage fluid. Empiric antibiotic treatment including clarithromycin was given, but the microbiological work-up remained negative. Serum IgE, C3, and C4 concentrations were normal. He recovered within 3 weeks after onset.We systematically reviewed 23 reports describing 30 additional cases of rituximab-associated lung disease. Twenty eight patients had received rituximab for B-cell malignancies, one for graft-versus host disease and one for immune thrombocytopenia. Median age was 64 years (interquartile range [IQR] 58-69 years). Seventy one percent received concomitant chemotherapy. Time to onset from the last rituximab dose was 14 days (IQR 11-22 days). Eleven of 31 patients required mechanical ventilation, and 9 died (29%). Ventilation was a significant predictor of fatal outcome (odds ratio 46.7; confidence interval 9.5-229.9). High dose glucocorticoid therapy did not improve survival or prevent severe lung disease or death. CONCLUSIONS: With the expanding use of rituximab for novel indications, additional cases of RALI affecting younger age groups are expected to emerge. Mechanical ventilation predicts poor outcome. Glucocorticoids may not be protective. PMID- 19681065 TI - Application of hyphenated mass spectrometry techniques for the analysis of urinary free glucocorticoids. AB - Alteration of levels of glucocorticoids in plasma and urine can be related to several diseases. In particular, the determination of endogenous glucocorticoids in urine has been reported to provide information on cortisol and cortisone status, on the activities of steroid hormone enzymes and on glucocorticoid metabolism. In this study, the application of hyphenated mass spectrometry techniques (GC/MS without derivatization and LC/MS) for the simultaneous analysis of free urinary cortisol (F), cortisone (E), tetrahydrocortisol (THF), allo tetrahydrocortisol (A-THF) and tetrahydrocortisone (THE) was evaluated. A sample preparation protocol by solid-phase extraction, mass spectrometry parameters and chromatographic conditions for both techniques were carefully optimized in terms of extracting phase and solvents, matrix effects, recovery, sensitivity and compound resolution. Baseline separation was achieved for the five underivatized analytes both in GC and LC. The LC/MS/MS technique was more suitable for the analysis of urine samples, being less influenced by matrix effects and showing excellent sensitivity and selectivity. A preliminary application of the reported method for the diagnosis of metabolic diseases was also described. The determination of each analyte in its free form, described for the first time in the paper, offers new perspectives in the application of glucocorticoid analysis for diagnostic purposes. PMID- 19681066 TI - Diastereomeric differentiation of norbornene amino acid peptides by electrospray ionization tandem mass spectrometry. AB - A new class of diastereomeric pairs of non-natural amino acid peptides derived from butyloxycarbonyl (Boc-)protected cis-(2S,3R)- and trans-(2S,3S)-beta norbornene amino acids including a monomeric pair have been investigated by electrospray ionization (ESI) tandem mass spectrometry using quadrupole time-of flight (Q-TOF) and ion-trap mass spectrometers. The protonated cis-BocN-beta-nbaa (2S,3R) (1) (betanbaa = beta-norbornene amino acid) eliminates the Boc group to form [M+H-Boc+H](+), whereas an additional ion [M+H-C(4)H(8)](+) is formed from trans-BocN-beta-nbaa (2S,3S) (2). Similarly, it is observed that the peptide diastereomers (di-, tri- and tetra-), with cis-BocN-beta-nbaa (2S,3R)- at the N terminus, initially eliminate the Boc group to form [M+H-Boc+H](+) which undergo further fragmentation to give a set of product ions that are different for the peptides with trans-BocN-beta-nbaa (2S,3S)- at the N-terminus. Thus the Boc group fragments differently depending on the configuration of the amino acid present at the N-terminus. It is also observed that the peptide bond cleavage in these peptides is less favoured and most of the product ions are formed due to retro Diels-Alder fragmentation. Interestingly, sodium-cationized peptide diastereomers mainly yield a series of retro-Diels-Alder fragment ions which are different for each diastereomer as they are formed starting from [M+Na-Boc+H](+) in peptides with cis-BocN-beta-nbaa (2S,3R)- at the N-terminus, and [M+Na-C(4)H(8)](+) in peptides with trans-BocN-beta-nbaa (2S,3S)- at the N-terminus. All these results clearly indicate that these diastereomeric pairs of peptides yield characteristic product ions which help distinguish the isomers. PMID- 19681064 TI - Beryllium lymphocyte proliferation test surveillance identifies clinically significant beryllium disease. AB - BACKGROUND: Workplace surveillance identifies chronic beryllium disease (CBD) but it remains unknown over what time frame mild CBD will progress to a more severe form. METHODS: We examined physiology and treatment in 229 beryllium sensitization (BeS) and 171 CBD surveillance-identified cases diagnosed from 1982 to 2002. Never smoking CBD cases (81) were compared to never smoking BeS patients (83) to assess disease progression. We compared CBD machinists to non-machinists to examine effects of exposure. RESULTS: At baseline, CBD and BeS cases did not differ significantly in exposure time or physiology. CBD patients were more likely to have machined beryllium. Of CBD cases, 19.3% went on to require oral immunosuppressive therapy. At 30 years from first exposure, measures of gas exchange were significantly worse and total lung capacity was lower for CBD subjects. Machinists had faster disease progression as measured by pulmonary function testing and gas exchange. CONCLUSIONS: Medical surveillance for CBD identifies individuals at significant risk of disease progression and impairment with sufficient time since first exposure. PMID- 19681068 TI - A novel multistep mechanism for the stereocontrolled ring opening of hindered sulfamidates: mild, green, and efficient reactivity with alcohols. AB - Cyclic hindered sulfamidates exhibited an outstanding performance in their ring opening reactions with alcohols and in the absence of any external activator. The mechanism of this unprecedented transformation was thoroughly studied both experimentally and theoretically. As a result, a nontrivial stepwise pathway involving solvent-induced conversion of the sulfamidates to activated aziridinium and then to oxazolinium cations, which are finally opened at their 5-position with inversion of configuration, is proposed. The presence of the SO(3) moiety in the sulfamidate was revealed as a "built-in activator". In fact, the spontaneous SO(3) cleavage takes place under the reaction conditions and avoids the subsequent step of hydrolysis after the ring opening of the sulfamidates. This is another important improvement of this methodology with respect to the standard basic conditions, allowing a greater compatibility with other functional groups. Furthermore, the carbamate group plays a key role in this mechanism. Briefly, a highly chemoselective and stereoespecific formal solvolysis of hindered sulfamidates with alcohols without further activation is described. This reaction takes place exclusively at the quaternary center with inversion of configuration, providing a new straightforward synthetic route to O-substituted alpha methylisoserines. PMID- 19681067 TI - S(N)2' reaction of organozinc reagents activated by catalytic tBu-P4 base in the presence of LiCl. AB - The tBu-P4 base was found to be an excellent catalyst for the activation of organozinc reagents. The base was used to promote the S(N)2' reaction of alpha,beta-unsaturated esters bearing a gamma-chloride in the reactions with various organozinc reagents in the presence of LiCl. The reactions proceeded in high yield with excellent chemo-and regioselectivity. The role of LiCl appears to be the activation of the gamma-chloride of the alpha,beta-unsaturated esters, which was confirmed by NMR spectroscopic study. PMID- 19681069 TI - Self-assembly of a chiral lipid gelator controlled by solvent and speed of gelation. AB - Glutamine derivative 1 with two-photon absorbing units has been synthesized and was found to show gelation ability in some solvents. Its self-assembly in the gel phase could be controlled by the solvent and speed of gelation. For example, in DMSO the organogelator self-assembled into H-aggregates with weak exciton coupling between the aromatic moieties. On the other hand, in DMSO/diphenyl ether (1:9, v/v) the molecules formed 1D aggregates, but with strong exciton coupling due to the small distance between the chromophores. Moreover, the formation of these two kinds of aggregates could be adjusted by the ratio of DMSO to diphenyl ether. In DMSO/toluene, DMSO/butanol, DMSO/butyl acetate, and DMSO/acetic acid systems similar results were observed. Therefore, conversion of the packing model occurs irrespective of the nature of the solvent. Notably, a unique sign inversion in the CD spectra could be realized by controlling the speed of gelation in the DMSO/diphenyl ether (1:9, v/v) system. It was found that a low speed of gelation induces the gelator to adopt a packing model with strong pi-pi interactions between the aromatic units. Moreover, the gels, when excited at 800 nm, emit strong green fluorescence and the quantum chemical calculations suggest that intramolecular charge transfer leads to two-photon absorption of the gelator molecule. PMID- 19681070 TI - Conditional deletion of the Lkb1 gene in the mouse mammary gland induces tumour formation. AB - Heterozygous germline mutations in the LKB1 (STK11) gene cause Peutz-Jeghers syndrome (PJS), an autosomal dominant disorder characterized by hamartomatous polyposis of the gastrointestinal tract and an increased risk of colorectal, breast, and other cancers. To model the role of LKB1 mutation in mammary tumourigenesis, we have used a conditional gene targeting strategy to generate a mouse in which exons encoding the kinase domain of Lkb1 were deleted specifically in the mammary gland. Mammary gland tumours developed in these mice with a latency of 46-85 weeks and occurred in the thoracic or inguinal glands. These tumours were grade 2 invasive ductal carcinomas or solid papillary carcinomas with histological features similar to those described in breast cancers arising in patients with PJS. This mouse model of Lkb1 deficiency provides a potentially useful tool to investigate the role of Lkb1 in tumourigenesis and to guide the development of therapeutic approaches. PMID- 19681071 TI - Tid1 functions as a tumour suppressor in head and neck squamous cell carcinoma. AB - Human tumourous imaginal disc (Tid1), a human homologue of the Drosophila tumour suppressor protein Tid56, is involved in multiple intracellular signalling pathways such as apoptosis, cell proliferation, and cell survival. Here, we investigated the anti-tumourigenic activity of Tid1 in head and neck squamous cell carcinoma (HNSCC) in vitro and in vivo. Firstly, the clinical association between Tid1 expression and progression of HNSCC was explored. It was found that expression of Tid1 was negatively associated with tumour status, recurrence, and survival prognosis using immunohistochemical analysis of primary HNSCC patient tumour tissue. Secondly, ectopic expression of Tid1 in HNSCC cells was shown to significantly inhibit cell proliferation, migration, invasion, anchorage independent growth, and xenotransplantation tumourigenicity. Thirdly, we showed that overexpression of Tid1 attenuated EGFR activity and blocked the activation of AKT in HNSCC cells, which are known to be involved in the regulation of survival in HNSCC cells. On the other hand, ectopic expression of constitutively active AKT greatly reduced apoptosis induced by Tid1 overexpression. Together, these findings suggest that Tid1 functions as a tumour suppressor in HNSCC tumourigenesis. PMID- 19681072 TI - Exploring the anion-cation interaction in m-terphenyltetrafluorosilicates by using multinuclear NMR spectroscopy, X-ray diffraction, and ICR-FT-MS. AB - The synthesis of a series of m-terphenyl-substituted tetrafluorosilicates with different cations (Na(+), K(+), Rb(+), Cs(+), Ag(+), Tl(+)) is described and the interactions between the anion and cation are investigated in the solid, solution, and gas states by using multinuclear NMR spectroscopy, X-ray diffraction, and ion cyclotron resonance Fourier-transform mass spectrometry (ICR FT-MS). In solution, heteronuclear NMR spectroscopy parameters show only limited sensitivity to the nature of the cation, which furthermore can be affected by solvent effects. More pronounced effects are observed in the structural data obtained from X-ray diffraction studies, which are in good agreement with experimental gas-phase data from ESIMS. ESIMS also reveals the existence of dimeric species of the type [M(DmpSiF(4))(2)](-) (Dmp = 2,6-dimesitylphenyl), the stability of which was determined by normalized collision energy experiments. PMID- 19681073 TI - Gold manno-glyconanoparticles: multivalent systems to block HIV-1 gp120 binding to the lectin DC-SIGN. AB - The HIV envelope glycoprotein gp120 takes advantage of the high-mannose clusters on its surface to target the C-type lectin dendritic cell-specific intracellular adhesion molecule-3-grabbing non-integrin (DC-SIGN) on dendritic cells. Mimicking the cluster presentation of oligomannosides on the virus surface is a strategy for designing carbohydrate-based antiviral agents. Bio-inspired by the cluster presentation of gp120, we have designed and prepared a small library of multivalent water-soluble gold glyconanoparticles (manno-GNPs) presenting truncated (oligo)mannosides of the high-mannose undecasaccharide Man(9)GlcNAc(2) and have tested them as inhibitors of DC-SIGN binding to gp120. These glyconanoparticles are ligands for DC-SIGN, which also interacts in the early steps of infection with a large number of pathogens through specific recognition of associated glycans. (Oligo)mannosides endowed with different spacers ending in thiol groups, which enable attachment of the glycoconjugates to the gold surface, have been prepared. manno-GNPs with different spacers and variable density of mannose (oligo)saccharides have been obtained and characterized. Surface plasmon resonance (SPR) experiments with selected manno-GNPs have been performed to study their inhibition potency towards DC-SIGN binding to gp120. The tested manno-GNPs completely inhibit the binding from the micro- to the nanomolar range, while the corresponding monovalent mannosides require millimolar concentrations. manno-GNPs containing the disaccharide Manalpha1-2Manalpha are the best inhibitors, showing more than 20 000-fold increased activity (100 % inhibition at 115 nM) compared to the corresponding monomeric disaccharide (100 % inhibition at 2.2 mM). Furthermore, increasing the density of dimannoside on the gold platform from 50 to 100 % does not improve the level of inhibition. PMID- 19681074 TI - 5-(1,2,3-Triazol-1-yl)tetrazole derivatives of an azidotetrazole via click chemistry. AB - N-C bonded (non-bridged) 5-(1,2,3-triazol-1-yl)tetrazoles were synthesized by the Cu(I)-catalyzed 1,3-dipolar azide-alkyne cycloaddition click reaction using 5 azido-N-(propan-2-ylidene)-1H-tetrazole (1). For example, the click reaction of 1 in the presence of CuSO(4)5 H(2)O and Na ascorbate at 65-70 degrees C for 48 h in CH(3)CN/H(2)O co-solvent was found to be limited to only terminal alkynes that have electron-withdrawing groups, CF(3)C[triple chemical bond]CH (2 a) and SF(5)C[triple chemical bond]CH (2 b), giving rise to isopropylidene-[5-(4 trifluoromethyl-1,2,3-triazol-1-yl)tetrazol-1-yl]amine (3 a) and isopropylidene [5-(4-pentafluorosulfanyl-1,2,3-triazol-1-yl)tetrazol-1-yl]amine (3 b) in 47 % and 66 % yields, respectively. When carried out under conditions using CuI and 2,6-lutidine as catalysts at 0 degrees C for 13 h in CHCl(3), the click reaction was versatile toward alkynes even those having electron-donating groups. Properties of new products were determined and compared with those of 1. Heats of formation, detonation pressures, detonation velocities and impact sensitivities are reported for these new 5-(1,2,3-triazol-1-yl)tetrazoles. PMID- 19681075 TI - Stereocontrolled and versatile total synthesis of bispyrrolidinoindoline diketopiperazine alkaloids: structural revision of the fungal isolate (+) asperdimin. AB - Homo- and heterodimeric bispyrrolidinoindoline diketopiperazine alkaloids have been synthesized following a concise, versatile, and stereoselective route. Highlights of the sequence are a diastereoselective construction of the C3a-bromo hexahydropyrrolo[2,3-b]indole nucleus, its Co(I)-induced C3a-C3a' dimerization, and the twofold or sequential amide-bond formation before cyclization to the diketopiperazine of the homo- or heterodimeric alkaloids, respectively. Stereochemical diversity is achieved through the choice of the appropriate amino acids combined with the base-induced epimerization of the C2-acyl hexahydropyrrolo[2,3-b]indole at C2. According to this strategy, the natural products (+)-WIN 64821 1, (+)-WIN 64745 2 and (+)-asperdimin 6 as well as analogues (5, 22, 32, 44) with different relative and absolute configuration have been efficiently synthesized. The flexibility of this synthetic methodology has facilitated the structural revision of the natural product (+)-asperdimin, whose structure has been corrected to diastereomer 6. PMID- 19681077 TI - Novel prolinamide-camphor-containing organocatalysts for direct asymmetric Michael addition of unmodified aldehydes to nitroalkenes. PMID- 19681076 TI - Hydrogen-deuterium exchange reactions of aromatic compounds and heterocycles by NaBD4-activated rhodium, platinum and palladium catalysts. AB - Conventional thermal and microwave conditions were compared for hydrogen deuterium (H/D) exchange reactions of aminobenzoic acids catalysed by NaBD(4) activated Pd/C or RhCl(3) with D(2)O as the deuterium source. We also investigated different NaBD(4)-activated metal catalysts (including Pd/C, RhCl(3) and Pt/C) under microwave conditions for an efficient H/D exchange of aromatic and heterocyclic compounds. Even higher deuterium incorporations were obtained for Pd/C and Pt/C catalyst mixtures due to the previously observed synergistic effect. Finally, we have applied these optimised conditions for one-step syntheses of the MS standards of several pharmaceutically active compounds. PMID- 19681078 TI - Anionic gold(I) complexes-twelve- and ten-vertex monocarba-closo-borate anions with carbon-gold sigma bonds. AB - The anionic gold(I) complexes [1-(Ph(3)PAu)-closo-1-CB(11)H(11)](-) (1), [1 (Ph(3)PAu)-closo-1-CB(9)H(9)](-) (2), and [2-(Ph(3)PAu)-closo-2-CB(9)H(9)](-) (3) with gold-carbon 2c-2e sigma bonds have been prepared from [AuCl(PPh(3))] and the respective carba-closo-borate dianion. The anions have been isolated as their Cs(+) salts and the corresponding [Et(4)N](+) salts were obtained by salt metathesis reactions. The salt Cs-3 isomerizes in the solid state and in solution at elevated temperatures to Cs-2 with DeltaH(iso)=(-75+/-5) kJ mol(-1) (solid state) and DeltaH( not equal)=(118+/-10) kJ mol(-1) (solution). The compounds were characterized by vibrational and multi-NMR spectroscopies, mass spectrometry, elemental analysis, and differential scanning calorimetry. The crystal structures of [Et(4)N]-1, [Et(4)N]-2, and [Et(4)N]-3 were determined. The bonding parameters, NMR chemical shifts, and the isomerization enthalpy of Cs-3 to Cs-2 are compared to theoretical data. PMID- 19681079 TI - White-light-emitting DNA (WED). PMID- 19681080 TI - Synthesis, structure, photoluminescence and photoreactivity of 2,3-diphenyl-4 neopentyl-1-silacyclobut-2-enes. AB - A series of six 2,3-diphenyl-4-neopentyl-1-silacyclobut-2-enes with different 1,1 substituents has been prepared and characterized by single-crystal X-ray crystallography. These compounds possess a cis-stilbene-like chromophore involving also the four-membered ring, and exhibit a photophysical behavior similar to that of previously reported 1,2-diphenyl-cycloalkenes. This chromophore system is confirmed by a theoretical investigation of the electronic structure and excitation spectra. The absorption and photoluminescence of selected derivatives were studied in solution, as solid powder samples, and in doped-polymer thin films. In well-dissolved solution, the silacyclobutenes show only very weak fluorescence emission (quantum yield approximately 0.1%), due to competition with photochemical and non-radiative photophysical relaxation. When the solubility is degraded in a poor (aqueous) solvent, the formation of nanoscale aggregates leads to a significant enhancement factor in the emission intensity, due to the suppression of the photoreactivity in the more rigid molecular environment, although the quantum yield still remains below a few percent. In the solid-state, however, photoreactivity is completely suppressed leading to fluorescence quantum efficiencies of 8-23% depending on the 1,1 substituents, which demonstrates these compounds' potential as chromophores for condensed-phase luminescence applications. Two dominant competing photochemical reactions have been identified in solution (for excitation in the lowest-energy absorption band, >260 nm), which are analogous to related (sila-)cyclobutenes and stilbenoids. The first involves ring-opening due to cleavage of the 1,4-Si-C bond to form metastable silabutadienes, which was confirmed by isolating the stereospecifically formed allylsilane which results from a secondary reaction with trapping agents such as methanol or water. The second photochemical reaction involves ring closure of the 2,3-diphenyl substructure to form a dihydrophenanthrene analogue, which was confirmed by isolating the phenanthrene derivative that results following subsequent hydrogen abstraction in the presence of oxygen. Measurements of the silacyclobutenes in doped-polymer thin films reveal a spectroscopic behavior ranging from that in solution to the nano aggregate case as the silacyclobutene dopant concentration is increased. PMID- 19681081 TI - Catalytic C-C, C-N, and C-O Ullmann-type coupling reactions. AB - Copper-catalyzed Ullmann condensations are key reactions for the formation of carbon-heteroatom and carbon-carbon bonds in organic synthesis. These reactions can lead to structural moieties that are prevalent in building blocks of active molecules in the life sciences and in many material precursors. An increasing number of publications have appeared concerning Ullmann-type intermolecular reactions for the coupling of aryl and vinyl halides with N, O, and C nucleophiles, and this Minireview highlights recent and major developments in this topic since 2004. PMID- 19681082 TI - A general and mild palladium-catalyzed domino reaction for the synthesis of 2H indazoles. PMID- 19681083 TI - Monitoring protein kinases in cellular media with highly selective chimeric reporters. PMID- 19681084 TI - A high-resolution interaction map of three transcriptional activation domains with a key coactivator from photo-cross-linking and multiplexed mass spectrometry. PMID- 19681086 TI - Polymer-functionalized platinum-on-gold bimetallic nanorods. PMID- 19681085 TI - Inclusion of anionic guests inside a molecular cage with palladium(II) centers as electrostatic anchors. PMID- 19681087 TI - Lipopeptides from Staphylococcus aureus as Tlr2 Ligands: prediction with mrna expression, chemical synthesis, and immunostimulatory activities. PMID- 19681088 TI - The effect of pressure on ZIF-8: increasing pore size with pressure and the formation of a high-pressure phase at 1.47 GPa. PMID- 19681089 TI - Pure samples of individual conformers: the separation of stereoisomers of complex molecules using electric fields. PMID- 19681090 TI - Controlling bacterial biofilms. PMID- 19681091 TI - Are the catalytic properties of enzymes from piezophilic organisms pressure adapted? AB - We report the crystal structure of dihydrofolate reductase (DHFR) from the psychropiezophilic bacterium Moritella profunda, which was isolated from the deep ocean at 2 degrees C and 280 bar. The structure is typical of a chromosomal DHFR and we were unable to identify any obvious structural features that would suggest pressure adaptation. In particular, the core regions of the enzyme are virtually identical to those of the DHFR from the mesophile Escherichia coli. The steady state rate at pH 9, which is limited by hydride transfer at atmospheric pressure, is roughly constant between 1 and 750 bar, falling at higher pressures. However, the value of K(M) increases with increasing pressure, and as a result k(cat)/K(M) decreases over the entire pressure range studied. Isotope effect studies showed that increasing the pressure causes a change in the rate-limiting step of the reaction. We therefore see no evidence of pressure adaptation in either the structure or the activity of this enzyme. PMID- 19681092 TI - Magnetic alignment in solid state and temperature hysteresis in aqueous tetrahydrofuran solution for tetrathiafulvaleno[18]annulenes. AB - Tris(tetrathiafulvaleno)dodecadehydro[18]annulene-hexaesters have a multi functionality that is very sensitive to small differences in the ester side chain. Self-aggregation of the [18]annulenes in amphiphilic media such as THF H(2)O (v/v, 1:1) either produce a fibrous structure or result in temperature hysteresis of the color and (1)H NMR signals. This temperature hysteresis in solution is due to both strong self-aggregation behavior and unique cluster formation in a binary solution of THF and water. PMID- 19681093 TI - Structure-Based Design, Synthesis, and Evaluation of 2'-(2-Hydroxyethyl)-2' deoxyadenosine and the 5'-Diphosphate Derivative as Ribonucleotide Reductase Inhibitors. AB - Analysis of the recently solved X-ray crystal structures of Saccharomyces cerevisiae ribonucleotide reductase I (ScRnr1) in complex with effectors and substrates led to the discovery of a conserved water molecule located at the active site that interacted with the 2'-hydroxy group of the nucleoside ribose. In this study 2'-(2-hydroxyethyl)-2'-deoxyadenosine 1 and the 5'-diphosphate derivative 2 were designed and synthesized to see if the conserved water molecule could be displaced by a hydroxymethylene group, to generate novel RNR inhibitors as potential antitumor agents. Herein we report the synthesis of analogues 1 and 2, and the co-crystal structure of adenosine diphosphate analogue 2 bound to ScRnr1, which shows the conserved water molecule is displaced as hypothesized. PMID- 19681094 TI - Up-regulation of autophagy in small intestine Paneth cells in response to total body gamma-irradiation. AB - Macroautophagy (mAG) is a lysosomal mechanism of degradation of cell self constituents damaged due to variety of stress factors, including ionizing irradiation. Activation of mAG requires expression of mAG protein Atg8 (LC3) and conversion of its form I (LC3-I) to form II (LC3-II), mediated by redox-sensitive Atg4 protease. We have demonstrated upregulation of this pathway in the innate host defense Paneth cells of the small intestine (SI) due to ionizing irradiation and correlation of this effect with induction of pro-oxidant inducible nitric oxide synthase (iNOS). CD2F1 mice were exposed to 9.25 Gy gamma-ionizing irradiation. Small intestinal specimens were collected during 7 days after ionizing irradiation. Assessment of ionizing irradiation-associated alterations in small intestinal crypt and villus cells and activation of the mAG pathway was conducted using microscopical and biochemical techniques. Analysis of iNOS protein and the associated formation of nitrites and lipid peroxidation products was performed using immunoblotting and biochemical analysis, and revealed increases in iNOS protein, nitrate levels and oxidative stress at day 1 following ionizing irradiation. Increase in immunoreactivity of LC3 protein in the crypt cells was observed at day 7 following ionizing irradiation. This effect predominantly occurred in the CD15-positive Paneth cells and was associated with accumulation of LC3-II isoform. The formation of autophagosomes in Paneth cells was confirmed by transmission electron microscopy (TEM). Up-regulation of LC3 pathway in the irradiated SI was accompanied by a decreased protein-protein interaction between LC3 and chaperone heat shock protein 70. A high-level of LC3 immunoreactivity in vacuole-shaped structures was spatially co-localized with immunoreactivity of 3-nitro-tyrosine. The observed effects were diminished in iNOS knockout B6.129P2-NOS2(tm1Lau)/J mice subjected to the same treatments. We postulate that the observed up-regulation of mAG in the irradiated small intestine is at least in part mediated by the iNOS signalling mechanism. PMID- 19681095 TI - Chronic exercise decreases cytokine production in healthy rat skeletal muscle. AB - Skeletal muscle is the source of pro- and anti-inflammatory cytokines, and recently, it has been recognized as an important source of interleukin-6 (IL-6). Acute physical exercise is known to induce a pro-inflammatory cytokine profile in the plasma. However, the effect of chronic physical exercise in the production of pro- and anti-inflammatory cytokines by the skeletal muscle has never been examined. We assessed IL-6, TNF-alpha, IL-1beta and IL-10 levels in the skeletal muscle of rats submitted to endurance training. Animals were randomly assigned to either a sedentary group (S, n = 7) or an endurance exercise trained group (T, n = 8). Trained rats ran on a treadmill for 5 days week(-1) for 8 weeks (60% VO(2max)). Detection of IL-6, TNF-alpha, IL-1beta and IL-10 protein expression was carried out by ELISA. We found decreased expression of IL-1beta, IL-6, TNF alpha and IL-10 (28%, 27%, 32% and 37%, respectively, p < 0.05) in the extensor digital longus (EDL) from T, when compared with S. In the soleus, IL-1beta, TNF alpha and IL-10 protein levels were similarly decreased (34%, 42% and 50%, respectively, p < 0.05) in T in relation to S, while IL-6 expression was not affected by the training protocol. In conclusion, exercise training induced decreased cytokine protein expression in the skeletal muscle. These data show that in healthy rats, 8-week moderate-intensity aerobic training down regulates skeletal muscle production of cytokines involved in the onset, maintenance and regulation of inflammation, and that the response is heterogeneous according to fibre composition. PMID- 19681096 TI - Isotopologue enrichment factors of N(2)O reduction in soils. AB - Isotopic signatures can be used to study sink and source processes of N(2)O, but the success of this approach is limited by insufficient knowledge on the isotope fractionation factors of the various reaction pathways. We investigated isotope enrichment factors of the N(2)O-to-N(2) step of denitrification (epsilon) in two arable soils, a silt-loam Haplic Luvisol and a sandy Gleyic Podzol. In addition to the epsilon of (18)O (epsilon(18O)) and of average (15)N (epsilon(bulk)), the epsilon of the (15)N site preference within the linear N(2)O molecule (epsilon(SP)) was also determined. Soils were anaerobically incubated in gas tight bottles with N(2)O added to the headspace to induce N(2)O reduction. Pre treatment included the removal of NO(3) (-) to prevent N(2)O production. Gas samples were collected regularly to determine the dynamics of N(2)O reduction, the time course of the isotopic signatures of residual N(2)O, and the associated isotope enrichment factors. To vary reduction rates and associated fractionation factors, several treatments were established including two levels of initial N(2)O concentration and anaerobic pre-incubation with or without addition of N(2)O. N(2)O reduction rates were affected by the soil type and initial N(2)O concentration. The epsilon(18O) and epsilon(bulk) ranged between -13 and -20 per thousand, and between -5 and -9 per thousand, respectively. Both quantities were more negative in the Gleyic Podzol. The epsilon of the central N position (epsilon(alpha)) was always larger than that of the peripheral N-position (epsilon(beta)), giving epsilon(SP) of -4 to -8 per thousand. The ranges and variation patterns of epsilon were comparable with those from previous static incubation studies with soils. Moreover, we found a relatively constant ratio between epsilon(18O) and epsilon(bulk) which is close to the default ratio of 2.5 that had been previously suggested. The fact that different soils exhibited comparable epsilon under certain conditions suggests that these values could serve to identify N(2)O reduction from the isotopic fingerprints of N(2)O emitted from any soil. PMID- 19681097 TI - Effects of substrate surfaces in matrix-assisted laser desorption/ionization mass spectrometry. AB - For matrix-assisted laser desorption/ionization (MALDI) mass spectra, undesirable ion contamination can occur due to the direct laser excitation of substrate materials (i.e., laser desorption/ionization (LDI)) if the samples do not completely cover the substrate surfaces. In this study, comparison is made of LDI processes on substrates of indium and silver, which easily emit their own ions upon laser irradiation, and conventional materials, stainless steel and gold. A simultaneous decrease of ion intensities with the number of laser pulses is observed as a common feature. By the application of an indium substrate to the MALDI mass spectrometry of alkali salts and alkylammonium salts mixed with matrices, 2,5-dihydroxybenzoic acid (DHB) or N-(4-methoxybenzylidene)-4 butylaniline (MBBA), the mixing of LDI processes can be detected by the presence of indium ions in the mass spectra. This method has also been found to be useful for investigating the intrinsic properties of the MALDI matrices: DHB samples show an increase in the abundance of fragment ions of matrix molecules and cesium ions with the number of laser pulses irradiating the same sample spot; MBBA samples reveal a decrease in the level of background noise with an increase in the thickness of the sample layer. PMID- 19681098 TI - Mass spectrometric analysis of peptides from an immobilized lipase: focus on oxidative modifications. AB - Liquid chromatography/tandem mass spectrometry (LC/MS/MS) was used to study the primary structure of immobilized Candida antarctica lipase B (Novozym(R)435) without detaching the enzyme from the carrier. The immobilized enzyme packed in a miniature column was subjected to proteolysis and the peptides released were injected into the mass spectrometer for analysis. The set-up was utilized to determine amino acid oxidation after treatment of the biocatalyst with hydrogen peroxide. In total, sequence coverage of more than 90% was obtained, containing almost all of the amino acids sensitive to oxidation. Oxidation of methionine, tryptophan and cystine residues was observed. The flow system also allowed evaluation of the enzyme activity prior to peptide analysis. The developed method is general and should be applicable to other immobilized enzyme systems and to different treatments. PMID- 19681099 TI - Two-injection workflow for a liquid chromatography/LTQ-Orbitrap system to complete in vivo biotransformation characterization: demonstration with buspirone metabolite identification. AB - The relatively high background matrix in in vivo samples typically poses difficulties in drug metabolite identification, and causes repeated analytical runs on unit resolution liquid chromatography/mass spectrometry (LC/MS) systems before the completion of biotransformation characterization. Ballpark parameter settings for the LTQ-Orbitrap are reported herein that enable complete in vivo metabolite identification within two HPLC/MS injections on the hybrid LTQ Orbitrap data collection system. By setting the FT survey full scan at 60K resolution to trigger five dependent LTQ MS(2) scans, and proper parameters of Repeat Duration, Exclusion Duration and Repeat Count for the first run (exploratory), the Orbitrap achieved the optimal parallel data acquisition capability and collected maximum number of product ion scans. Biotransformation knowledge based prediction played the key role in exact mass ion extraction and multiple mass defect filtration when the initial data was processed. Meanwhile, product ion extraction and neutral loss extraction of the initial dependent data provided additional bonus in identifying metabolites. With updated parent mass list and the data-dependent setting to let only the ions on the parent mass list trigger dependent scans, the second run (confirmatory) ensures that all precursor ions of identified metabolites trigger not only dependent product ion scans, but also at or close to the highest concentration of the eluted metabolite peaks. This workflow has been developed for metabolite identification of in vivo or ADME studies, of which the samples typically contain a high level of complex matrix. However, due to the proprietary nature of the in vivo studies, this workflow is presented herein with in vitro buspirone sample incubated with human liver microsomes (HLM). The major HLM-mediated biotransformation on buspirone was identified as oxidation or hydroxylation since five mono- (+16 Da), seven di- (+32 Da) and at least three tri-oxygenated (+48 Da) metabolites were identified. Besides the metabolites 1-pyrimidinylpiperazine (1-PP) and hydroxylated 1-PP that formed by N-dealkylation, a new metabolite M308 was identified as the result of a second N-dealkylation of the pyrimidine unit. Two new metabolites containing the 8-butyl-8-azaspiro[4,5]decane-7,9-dione partial structure, M240 and M254, were also identified that were formed apparently due to the first N-dealkylation of the 1-PP moiety. PMID- 19681100 TI - Experimental determination of a Viviparus contectus thermometry equation. AB - Experimental measurements of the (18)O/(16)O isotope fractionation between the biogenic aragonite of Viviparus contectus (Gastropoda) and its host freshwater were undertaken to generate a species-specific thermometry equation. The temperature dependence of the fractionation factor and the relationship between Deltadelta(18)O (delta(18)O(carb.) - delta(18)O(water)) and temperature were calculated from specimens maintained under laboratory and field (collection and cage) conditions. The field specimens were grown (Somerset, UK) between August 2007 and August 2008, with water samples and temperature measurements taken monthly. Specimens grown in the laboratory experiment were maintained under constant temperatures (15 degrees C, 20 degrees C and 25 degrees C) with water samples collected weekly. Application of a linear regression to the datasets indicated that the gradients of all three experiments were within experimental error of each other (+/-2 times the standard error); therefore, a combined (laboratory and field data) correlation could be applied. The relationship between Deltadelta(18)O (delta(18)O(carb.) - delta(18)O(water)) and temperature (T) for this combined dataset is given by: T = - 7.43( + 0.87, - 1.13)*Deltadelta18O + 22.89(+/- 2.09) (T is in degrees C, delta(18)O(carb.) is with respect to Vienna Pee Dee Belemnite (VPDB) and delta(18)O(water) is with respect to Vienna Standard Mean Ocean Water (VSMOW). Quoted errors are 2 times standard error).Comparisons made with existing aragonitic thermometry equations reveal that the linear regression for the combined Viviparus contectus equation is within 2 times the standard error of previously reported aragonitic thermometry equations. This suggests there are no species-specific vital effects for Viviparus contectus. Seasonal delta(18)O(carb.) profiles from specimens retrieved from the field cage experiment indicate that during shell secretion the delta(18)O(carb.) of the shell carbonate is not influenced by size, sex or whether females contained eggs or juveniles. PMID- 19681101 TI - H2O, H2, HF, F2 and F2O nuclear magnetic shielding constants and indirect nuclear spin-spin coupling constants (SSCCs) in the BHandH/pcJ-n and BHandH/XZP Kohn-Sham limits. AB - Good performance of segmented contracted basis sets XZP, where X = D, T, Q and 5, for obtaining H(2)O, H(2), HF, F(2) and F(2)O nuclear isotropic shielding constants in the BHandH Kohn-Sham basis set limit was shown. The results of two- and three-parameter complete basis set limit extrapolation schemes were compared with experimental results, earlier literature data and benchmark ab initio results. Similar convergence patterns of shieldings obtained from calculations using general purpose XZP basis sets and from polarization-consistent basis sets pcS-n and pcJ-n, where n = 0, 1, 2, 3 and 4, designed to accurately predict magnetic properties were observed. On the contrary, the SSCCs were more sensitive to the XZP basis set size and generally less accurate than those estimated using pcJ-n basis set family. The BHandH density functional markedly outperforms B3LYP method in predicting heavy atom shieldings and SSCCs values in the studied systems. PMID- 19681102 TI - Acute stroke therapy with tissue plasminogen activator (tPA) since it was approved by the U.S. Food and Drug Administration (FDA). AB - Tissue plasminogen activator (tPA) for acute ischemic stroke was approved by the U.S. Food and Drug Administration (FDA) in 1996. Since then it has been severely underutilized. At the time when most practitioners were first being exposed to the literature concerning tPA, there were many concerns about safety and the restrictions on use were quite onerous. Since then a good deal of further work has been done to loosen the restrictions and allay concerns about the risks. The true risk to benefit ratio is far better than is generally realized. Now it is mostly economic problems related to the costs of constantly supplying emergency care that is limiting access. Furthermore, in the current litigious environment, failure to treat is likely to be a more hazardous course of action than legal exposure due to poor outcomes. It must be emphasized that the drug is quite safe and highly effective, and current utilization rates are unacceptably low. Ann Neurol 2009;66:6-10. PMID- 19681105 TI - Journeys in complex genetics: music and mental illness. PMID- 19681106 TI - Fusion processing of itraconazole solid dispersions by kinetisol dispersing: a comparative study to hot melt extrusion. AB - KinetiSol Dispersing (KSD) is a novel high energy manufacturing process investigated here for the production of pharmaceutical solid dispersions. Solid dispersions of itraconazole (ITZ) and hypromellose were produced by KSD and compared to identical formulations produced by hot melt extrusion (HME). Materials were characterized for solid state properties by modulated differential scanning calorimetry and X-ray diffraction. Dissolution behavior was studied under supersaturated conditions. Oral bioavailability was determined using a Sprague-Dawley rat model. Results showed that KSD was able to produce amorphous solid dispersions in under 15 s while production by HME required over 300 s. Dispersions produced by KSD exhibited single phase solid state behavior indicated by a single glass transition temperature (T(g)) whereas compositions produced by HME exhibited two T(g)s. Increased dissolution rates for compositions manufactured by KSD were also observed compared to HME processed material. Near complete supersaturation was observed for solid dispersions produced by either manufacturing processes. Oral bioavailability from both processes showed enhanced AUC compared to crystalline ITZ. Based on the results presented from this study, KSD was shown to be a viable manufacturing process for the production of pharmaceutical solid dispersions, providing benefits over conventional techniques including: enhanced mixing for improved homogeneity and reduced processing times. PMID- 19681107 TI - Mindfulness and attention deficit hyperactivity disorder. AB - Attention deficit hyperactivity disorder (ADHD) is a disorder characterized by attentional difficulties. Mindfulness is a receptive attention to present experience. Both ADHD and mindfulness are associated with attention and personality. This study tests whether individuals with ADHD have lower mindfulness scores than controls and, if true, whether personality contributes to these differences. One hundred and five adults (half with ADHD) were assessed for mindfulness, using the Kentucky Inventory of Mindfulness Skills, and personality, using the Tridimensional Character Inventory. Individuals with ADHD report themselves as less mindful than non-ADHD controls and more novelty-seeking, less self-directed, and more self-transcendent. Mindfulness is negatively associated with ADHD and positively associated with self-directedness and self transcendence. Analyses of subscales of mindfulness suggest that ADHD is associated most with the "Acting in Awareness" dimension, perhaps because of shared items reflecting attentional variability. The current findings support that a large portion of variability in trait mindfulness can be explained by ADHD status and personality traits of self-directedness and self-transcendence. It further suggests that interventions that increase mindfulness might improve symptoms of ADHD and increase self-directedness and/or self-transcendence. PMID- 19681108 TI - DeltaNp63 knockdown mice: A mouse model for AEC syndrome. AB - Dominant mutations in TP63 cause ankyloblepharon ectodermal dysplasia and clefting (AEC), an ectodermal dysplasia characterized by skin fragility. Since DeltaNp63alpha is the predominantly expressed TP63 isoform in postnatal skin, we hypothesized that mutant DeltaNp63alpha proteins are primarily responsible for skin fragility in AEC patients. We found that mutant DeltaNp63alpha proteins expressed in AEC patients function as dominant-negative molecules, suggesting that the human AEC skin phenotype could be mimicked in mouse skin by downregulating DeltaNp63alpha. Indeed, downregulating DeltaNp63 expression in mouse epidermis caused severe skin erosions, which resembled lesions that develop in AEC patients. In both cases, lesions were characterized by suprabasal epidermal proliferation, delayed terminal differentiation, and basement membrane abnormalities. By failing to provide structural stability to the epidermis, these defects likely contribute to the observed skin fragility. The development of a mouse model for AEC will allow us to further unravel the genetic pathways that are normally regulated by DeltaNp63 and that may be perturbed in AEC patients. Ultimately, these studies will not only contribute to our understanding of the molecular mechanisms that cause skin fragility in AEC patients, but may also result in the identification of targets for novel therapeutic approaches aimed at treating skin erosions. (c) 2009 Wiley-Liss, Inc. PMID- 19681112 TI - Preventable deaths, never events, and comparative effectiveness: it is time for US cardiologist to switch to transradial. PMID- 19681113 TI - 30-day risk prediction after coronary angioplasty: first step in decision making. PMID- 19681114 TI - Now, I am proud to be a Radialist. PMID- 19681115 TI - Ten years after: "I'd love to change the world". PMID- 19681116 TI - Percutaneous coronary intervention and 30-day mortality: the British Columbia PCI risk score. AB - OBJECTIVES: To construct a calculator to assess the risk of 30-day mortality following PCI. BACKGROUND: Predictors of 30-day mortality are commonly used to aid management decisions for cardiac surgical patients. There is a need for an equivalent risk-score for 30-day mortality for percutaneous coronary intervention (PCI) as many patients are suitable for both procedures. METHODS: The British Columbia Cardiac Registry (BCCR) is a population-based registry that collects information on all PCI procedures performed in British Columbia (BC). We used data from the BCCR to identify risk factors for mortality in PCI patients and construct a calculator that predicts 30-day mortality. RESULTS: Patients (total n = 32,899) were divided into a training set (n = 26,350, PCI between 2000 and 2004) and validation set (n = 6,549, PCI in 2005). Univariate predictors of mortality were identified. Multivariable logistic regression analysis was performed on the training set to develop a statistical model for prediction of 30 day mortality. This model was tested in the validation set. Variables that were objective and available before PCI were included in the final risk score calculator. The 30-day mortality for the overall population was 1.5% (n = 500). Area under the ROC curve was 90.2% for the training set and 91.1% for the validation set indicating that the model also performed well in this group. CONCLUSIONS: We describe a large, contemporary cohort of patients undergoing PCI with complete follow-up for 30-day mortality. A robust, validated model of 30-day mortality after PCI was used to construct a risk calculator, the BC-PCI risk score, which can be accessed at www.bcpci.org. PMID- 19681117 TI - "Drip-and-ship," "Stay-and-pray," "Freight-and-wait (and possibly inflate)" or simply opening the window wider?: Decision-making regarding time delays in the treatment of ST elevation MI. PMID- 19681118 TI - A new option for the "no-option" patient with refractory angina? PMID- 19681119 TI - RAS signaling dysregulation in human embryonal Rhabdomyosarcoma. AB - Rhabdomyosarcoma (RMS) is a common childhood solid tumor, resulting from dysregulation of the skeletal myogenesis program. Two major histological subtypes occur in childhood RMS, embryonal and alveolar. While chromosomal rearrangements account for the majority of alveolar tumors, the genetic defects underlying the pathogenesis of embryonal RMS remain largely undetermined. A few studies performed on small series of embryonal tumors suggest that dysregulation of RAS function may be relevant to disease pathogenesis. To explore further the biological and clinical relevance of mutations with perturbing consequences on RAS signaling in embryonal RMS, we investigated the prevalence of PTPN11, HRAS, KRAS, NRAS, BRAF, MEK1, and MEK2 mutations in a relatively large cohort of primary tumors. While HRAS and KRAS were found to be rarely mutated, we identified somatic NRAS lesions in 20% of cases. All mutations were missense and affected codon 61, with the introduction of a positive charged amino acid residue representing the most common event. PTPN11 was found mutated in one tumor specimen, confirming that somatic defects in this gene are relatively uncommon in RMS, while no mutation was observed in BRAF and MEK genes. Although no clear association of mutations with any clinical variable was observed, comparison of the outcome between mutation-positive and mutation-negative cases indicated a trend for a higher percentage of patients exhibiting a better outcome in the former. Our findings provide evidence that dysregulation of RAS signaling is a major event contributing to embryonal RMS pathogenesis. PMID- 19681120 TI - Protocadherin PCDH10, involved in tumor progression, is a frequent and early target of promoter hypermethylation in cervical cancer. AB - Cervical cancer (CC) is the second most common cancer in women. Currently, no tractable molecular-based therapeutic targets exist for patients with invasive CC and no predictive markers of risk assessment for progression of precancerous lesions are identified. New molecular insights into CC pathogenesis are urgently needed. Towards this goal, we first determined the copy number alterations of chromosome 4 and then examined the role of PCDH10 mapped to 4q28 as a candidate tumor suppressor gene. We identified monosomy 4 in 47% of 81 invasive CC studied by SNP array and found that 91% of 130 invasive CC harboring methylation in the promoter region of the PCDH10 gene. We then showed that aberrant promoter hypermethylation of PCDH10 is associated with downregulation of gene expression and cell lines exposed to demethylating agent resulted in profound reactivated gene expression. We also showed that the promoter methylation in the PCDH10 gene occurs at an earliest identifiable stage of low-grade squamous intraepithelial lesion. Our studies demonstrate that inactivation of PCDH10 may be a critical event in CC progression and form a potentially useful therapeutic target for CC. PMID- 19681121 TI - The shared tumor associated antigen cyclin-A2 is recognized by high-avidity T cells. AB - Cyclin-A2, a key cell cycle regulator, has been shown to be overexpressed in various types of malignancies with little expression in normal tissue. Such tumor associated genes potentially are useful targets for cancer immunotherapy. However, high-avidity cyclin-specific T cells are considered to be thymically deleted. We identified at least one nonameric HLA-A*0201 binding cyclin-A2 epitope by a reverse immunology approach. Using a highly efficient T-cell expansion system that is based on CD40-activated B (CD40-B) cells as sole antigen presenting cells we successfully generated cyclin-A2 specific CTL from HLA A*0201(+) donors. Interestingly, high-avidity cyclin-A2 specific CTL lines, which recognized peptide-pulsed and antigen expressing target cells, were indeed generated by stimulation with CD40-B cells when pulsed with low concentrations of peptide, whereas CD40-B cells pulsed at saturating concentrations could only induce low-avidity CTL, which recognized peptide-pulsed target cells only. One high-avidity CTL line was subcloned and CTL clones, whose peptide concentration required for half-maximal lysis were less than 1 nM, could lyse cyclin-A2 expressing tumor cells. Taken together, cyclin A2 is an attractive candidate for immune intervention in a significant number of cancer patients and high-avidity T cells can be readily generated using CD40-B cells as antigen-presenting cells. PMID- 19681122 TI - Oldies but goodies. PMID- 19681123 TI - Importance of creativity: new applications of the Proxis catheter. PMID- 19681124 TI - EGFR and KRAS status of primary sarcomatoid carcinomas of the lung: implications for anti-EGFR treatment of a rare lung malignancy. AB - Sarcomatoid carcinomas (SC) of the lung are uncommon malignant tumors composed of carcinomatous and sarcomatous cell components and characterized by a more aggressive outcome than other histological subtypes of nonsmall cell lung cancer (NSCLC). Although epidermal growth factor receptor (EGFR)-targeted therapies have emerged as a promising therapeutic approach in patients with advanced typical NSCLC such as adenocarcinoma, the potential clinical activity of these drugs in lung SC is still unknown. To investigate this point, we have analyzed the status of 4 EGFR pathways biomarkers in a series of lung SC. EGFR protein expression, EGFR gene copy number, EGFR mutational status and KRAS mutational status were assessed in a series of 22 consecutive cases of primary lung SC. EGFR protein overexpression was observed in all the cases. High level of polysomy (>or=4 copies of the gene in >40% of cells) was detected in 5 cases (23%). No EGFR mutation was detected. KRAS mutations were found in 8 patients (38%; Gly12Cys in 6 cases and Gly12Val in 2 cases). The consistent EGFR protein overexpression and the high rate of KRAS mutation may contribute to the poorer outcome of lung SC in comparison with typical NSCLC. The rare incidence of increased EGFR gene copy number, the lack of EGFR mutation and the high rate of KRAS mutation observed in our series also suggest that most patients with lung SC are not likely to benefit from anti-EGFR therapies. PMID- 19681125 TI - Novel method for delivering the Amplatzer muscular VSD occluder in a patient with double outlet right ventricle after bidirectional Glenn procedure and pulmonary artery band. AB - We report the first use of bilateral femoral venovenous rail creation for the delivery of an Amplatzer Muscular Ventricular Septal Defect Occluder in a patient with a large mid-to-apical muscular ventricular septal defect before Rastelli operation. The presence of a right-sided bidirectional Glenn shunt, a banded main pulmonary artery, and double outlet right ventricle anatomy precluded the use of standard delivery techniques. The patient underwent successful transcatheter device placement followed by Rastelli operation on the following day. PMID- 19681126 TI - Multiple device implantation in coronary sinus for functional mitral regurgitation: which is first? PMID- 19681127 TI - Trans-radial treatment of in-stent restenosis of the superficial femoral artery- the importance of case selection. PMID- 19681128 TI - Dermatologic findings of ankyloblepharon-ectodermal defects-cleft lip/palate (AEC) syndrome. AB - Hay-Wells syndrome, caused by mutations in the p63 gene, is an autosomal dominant ectodermal dysplasia with the main features of ankyloblepharon filiforme adnatum, ectodermal defects, and cleft lip/palate, from which the disorder's other name, AEC syndrome, is derived. The National Foundation for Ectodermal Dysplasias convened the International Research Symposium for AEC Syndrome on November 8-10, 2006, at Texas Children's Hospital/Baylor College of Medicine, Houston, TX with appropriate IRB approval. This multidisciplinary conference was the largest gathering of such patients to date and allowed us to further characterize dermatologic features of AEC syndrome, which included: sparse and wiry hair, nail changes, past or present scalp erosions, decreased sweat production, palmar/plantar changes, and unique pigmentary anomalies. Early recognition of the features of AEC syndrome and subsequent early diagnosis is important in minimizing invasive diagnostic studies, improving morbidity and mortality, and providing genetic counseling. Skin erosions, especially those of the scalp, were identified as the most challenging cutaneous aspect of this syndrome. Although the reasons for the skin erosions and poor healing are not known, mutations of p63 may lead to a diminished store of basal cells capable of replenishing the disrupted barrier. Therapeutic strategies currently under exploration include gene therapy, as well as epidermal stem cell therapy. Until then, gentle wound care and limiting further trauma seem to be the most prudent treatment modalities. PMID- 19681129 TI - Gene therapy and protein therapy of ectodermal dysplasias: a perspective. AB - Ectodermal dysplasias (EDs) form a complex and heterogeneous group of diseases currently defined and classified according to their clinical symptoms. The characterization, for several EDs, of the molecular events underlying their development, not only challenges this classification but also opens the door to new therapeutic options such as gene or protein therapy. This article provides a concise overview of the most recent successes and failures of this new type of treatment and sets in perspective how the specificities of given EDs will influence their feasibility in the near future. It makes the case for the need of new classification of EDs that is based on our most recent knowledge of the molecular basis of these diseases. PMID- 19681130 TI - Perspective on the classification of ectodermal dysplasia. AB - The need for revisiting the classification of ectodermal dysplasia syndromes has come. Prior to considering a new way to classify this group of disorders, however, thought should be given to some basic ideas about terminology and the process of classification. Consequently, this article reiterates the meanings of the words ectoderm, dysplasia, genetic and syndrome, and describes the process by which numeric taxonomists go about classification. In the process, a new family with the Jorgenson syndrome is described and the inheritance of the Schopf syndrome is clarified. Finally, the backgrounds of those with vested interests in the ectodermal dysplasia syndromes are described in order that all approaches to classification are covered. PMID- 19681131 TI - Towards a unified classification of the ectodermal dysplasias: opportunities outweigh challenges. AB - The ectodermal dysplasias include a complex and highly diverse group of heritable disorders that share in common developmental abnormalities of ectodermal derivatives. The broader definition of ectodermal dysplasias (as heritable disorders involving at least two of the ectodermal derivatives nails, teeth, hair, and eccrine sweat glands) encompasses 170-200 conditions. Some conditions included by this definition are relatively common; others are rare and, in some cases, family-specific. Classification of the ectodermal dysplasias has largely been approached by categorizing patterns of clinical findings (phenotypic grouping). In the last 2 decades great progress has been made in understanding the molecular pathogenesis and inter-relatedness of some of these conditions and a new consensus approach to classification that incorporates this new information is needed. A comprehensive and definitive classification of these disorders would be highly valuable for the many stakeholders in ED. As disease-specific molecular treatments are developed, accurate classification will assume greater importance in designing registries to enable rapid identification of those with rare disorders who may wish to participate in clinical trials. Ideally a working classification of such a disparate collection of conditions would have a design and architecture that would facilitate easy accessibility by each of the key stakeholder groups and would encourage enhanced interaction between these parties. Attaining this objective is a major challenge but is achievable. This article reviews the historical-clinical perspective and the impact of recent developments in molecular biology in the field. Reflections are offered as to the future direction of classification systems in these disorders. PMID- 19681132 TI - Molecular aspects of hypohidrotic ectodermal dysplasia. AB - Hypohidrotic (anhidrotic) ectodermal dysplasia (HED) is a congenital syndrome characterized by sparse hair, oligodontia, and reduced sweating. It is caused by mutations in any of the three Eda pathway genes: ectodysplasin (Eda), Edar, and Edaradd which encode a ligand, a receptor, and an intracellular signal mediator of a single linear pathway, respectively. In rare cases, HED is associated with immune deficiency caused by mutations in further downstream components of the Eda pathway that are necessary for the activation of the transcription factor NF kappaB. Here I present a brief research update on the molecular aspects of this evolutionarily conserved pathway. The developmental role of Eda will be discussed in light of loss- and gain-of-function mouse models with emphasis on the past few years. PMID- 19681133 TI - What the future holds for ectodermal dysplasias: future research and treatment directions. AB - A contrarian view suggests that the ectodermal dysplasias, including more than 200 different disorders, represent clinical variability and molecular heterogeneity as well as complex multigene heritable conditions often characterized by dysmorphogenesis of derivatives of embryonic ectoderm and beyond. Controversy exists over which syndromes do or do not belong in the classification of the clinical features that characterize ectodermal dysplasias. For example, Ellis-van Creveld syndrome is characterized by abnormalities of the teeth and hair, as well as of the skeleton and the cardiovascular system. Precision in diagnosis often is a preamble for improved patient diagnosis, treatment and desired outcomes. In tandem, molecular studies of complex epithelial-mesenchymal interactions required for ectodermal derivatives (e.g., hair, nail, skin, teeth, and exocrine glands) continue to identify and explain many signal transduction pathways and networks related to ectodermal dysplasias. Meanwhile, major international investments in fundamental biomedical research continue to yield significant benefits to the larger society. The convergence of informatics, nanotechnology, genomics, and epigenetic studies with clinical medicine and dentistry promise major progress for special needs patients such as ectodermal dysplasias. For example, investments in the molecular biology of genes and their regulation and function now provide more than 30 candidates for specific biomarkers to improve diagnosis, prognosis, treatments, therapeutics, and biomaterials for ectodermal dysplasias. Innovations in high throughput genotyping, gene mapping, single nucleotide polymorphisms (SNPs), interference RNA treatments, bioimaging, tissue engineering and related biomimetic approaches to design and fabricate biomaterials, offer enormous promise for the future of ectodermal dysplasias. PMID- 19681135 TI - A novel missense mutation in SCYL1BP1 produces geroderma osteodysplastica phenotype indistinguishable from that caused by nullimorphic mutations. AB - Effect of aging on decreased skin elasticity and bone mass is well known. Geroderma osteodysplastica (GO) is a very rare autosomal recessive disorder that recapitulates these two phenotypes at a much younger age. Using homozygosity mapping and linkage analysis in four Saudi families we have identified two mutations in SCYL1BP1, consistent with the very recent report by Hennies et al. [Hennies et al. (2008); Nat Genet 40: 1410-1412]. Interestingly, the missense mutation identified in our study is associated with an identical phenotype to that seen with the other null mutations including the other mutation in this study. Our study, therefore, supplements the limited available data on SCYL1BP1 and further establishes deficiency of this recently described golgin as the only known cause of GO. PMID- 19681138 TI - Approach towards a new classification for ectodermal dysplasias: integration of the clinical and molecular knowledge. AB - Rapid advances in new basic knowledge have led to a greater understanding of the genetic, embryologic, molecular biologic, and functional mechanisms underlying the ectodermal dysplasias. Scientists, researchers, and clinicians from diverse fields desire a classification that meets the needs of these rapidly changing concepts which, in addition to being clinically useful, can help further advances in understanding the basic underlying mechanisms, and can also spur the development of new therapies. The currently used classification was designed for clinical utility and has not easily integrated with the new concepts. Various users have suggested that a new classification should be based on underlying genetic and molecular abnormality, functional mechanism, embryology, or clinical features. The goal of this conference was to advance towards a consensus and this manuscript is a summary of a workshop focused on an approach towards the development of a classification that integrates the clinical and molecular knowledge. PMID- 19681134 TI - Targeted deletion of Dicer disrupts lens morphogenesis, corneal epithelium stratification, and whole eye development. AB - Dicer, a ribonuclease essential for miRNA processing, is expressed abundantly in developing mouse cornea and lens. We studied the roles of Dicer and miRNAs in eye development by conditionally deleting the Dicer gene in the mouse lens and corneal epithelium. Adult Dicer conditional null (DicerCN) mice had severe microphthalmia with no discernible lens and a poorly stratified corneal epithelium. Targeted deletion of Dicer effectively inhibited miRNA processing in the developing lens at 12.5 day of embryogenesis (E12.5). Lens development initiated normally but underwent progressive dystrophy between E14.5 and E18.5. Microarray analysis revealed activation of P53 signaling in DicerCN lenses at E13.5, consistent with increased apoptosis and reduced cell proliferation between E12.5 and E14.5. Expression of Pax6 and other lens developmental transcription factors were not greatly affected between E12.5 and E14.5 but decreased as the lens degenerated. Our data indicated an indispensible role for Dicer and miRNAs in lens and corneal development. PMID- 19681140 TI - Fight the stupids! PMID- 19681141 TI - Nervecenter: consumer gene tests face uncertain legal, regulatory future. PMID- 19681142 TI - Oral findings in ankyloblepharon-ectodermal dysplasia-cleft lip/palate (AEC) syndrome. AB - This article outlines the dental conditions and treatment findings of individuals ranging in age from 4 months to 30 years of age diagnosed with ankyloblepharon ectodermal dysplasia-cleft lip/palate (AEC) syndrome. The average number of permanent teeth present is 4.75 with a range of 0-12 teeth. The most frequently present permanent teeth are first molars, canines and maxillary incisors. The dentition of AEC-affected individuals in our study is similar to reports in other ectodermal dysplasia syndromes. The dental treatment findings, including those necessitated by the cleft palate defect seen in the syndrome, are also similar to age appropriate treatment seen in other cases of ectodermal dysplasia syndromes. Only 22% of subjects had prosthetic replacement of missing teeth. History indicated that issues related to treatment of the palatal cleft and lack of recognition of the oral defects as a part of the underlying medical problem were factors in receiving comprehensive tooth replacement care. PMID- 19681143 TI - Diet composition, food intake, body condition, and fecal consistency in captive tapirs (Tapirus spp.) in UK collections. AB - Intake measurements were carried out in 22 tapirs from seven UK zoological collections. Dry matter intake (DMI) ranged from 48 to 86 g/kg(0.75)/d. Across collections, the highest proportion of the ingested diet consisted of pelleted feeds (including grains and bread) at 46+/-17% DMI, followed by commercial produce at 26+/-12% DMI, roughage (excluding browse) at 17+/-11% DMI, and browse at 11+/-11% DMI. The proportion of roughage, crude protein, crude fiber, and neutral detergent fiber levels in the diets investigated were well below levels recommended for domestic horses and other ungulates. Intakes of digestible energy (DE) as estimated from food nutrients using of a standard equation for domestic horses ranged from 0.58 to 0.88 MJ/DE/kg(0.75)/d, with many individuals exceeding the assumed maintenance requirement of 0.6 MJ/DE/kg(0.75)/d for hindgut fermenters. At values exceeding this DE intake, animals had higher than ideal body condition scores (BCS). Animals with higher BCS (i.e. more obese animals) generally had higher fecal scores (FS) (i.e. softer feces), and both BCS and FS were positively correlated to DMI and calculated DE intake. This suggests that the population studied was generally overfed, with resulting obesity and softer fecal consistency. The use of highly digestible feeds such as commercial produce and pelleted feeds should be restricted in the diets of these animals and roughage intake promoted in order to attempt to achieve normal BCS and FS in this captive population. PMID- 19681144 TI - Variation in visitor perceptions of a polar bear enclosure based on the presence of natural vs. un-natural enrichment items. AB - A debate exists among modern zoo staff as to whether or not the addition of un naturalistic enrichment takes away from, or even defeats, the educational messages designers are trying to incorporate in naturalistic exhibits. A visitor study was conducted at the Central Park Zoo's polar bear exhibit in order to determine whether or not the type of enrichment in an enclosure actually alters guest perceptions. Visitors were exposed to one of two enrichment treatments in the bear enclosure: Naturalistic or Un-naturalistic. The results of this study suggest that enrichment type did not alter the perceptions of visitors. However, it did identify some of the different ways adults and youths perceive animals and zoos. Additionally, the study highlighted the varying perceptions individuals have of the concept of polar bears vs. their perceptions of the captive individuals at the Central Park Zoo. Implications for enrichment usage and exhibit design are discussed. PMID- 19681139 TI - Gene expression profiles of lens regeneration and development in Xenopus laevis. AB - Seven hundred and thirty-four unique genes were recovered from a cDNA library enriched for genes up-regulated during the process of lens regeneration in the frog Xenopus laevis. The sequences represent transcription factors, proteins involved in RNA synthesis/processing, components of prominent cell signaling pathways, genes involved in protein processing, transport, and degradation (e.g., the ubiquitin/proteasome pathway), matrix metalloproteases (MMPs), as well as many other proteins. The findings implicate specific signal transduction pathways in the process of lens regeneration, including the FGF, TGF-beta, MAPK, Retinoic acid, Wnt, and hedgehog signaling pathways, which are known to play important roles in eye/lens development and regeneration in various systems. In situ hybridization revealed that the majority of genes recovered are expressed during embryogenesis, including in eye tissues. Several novel genes specifically expressed in lenses were identified. The suite of genes was compared to those up regulated in other regenerating tissues/organisms, and a small degree of overlap was detected. PMID- 19681145 TI - Biochemical and physiological observations in meerkats (Suricata suricatta) at two zoos during a dietary transition to a diet designed for insectivores. AB - This research (involving two separate institutions) assessed the serum chemistries and body weights of meerkats (Suricata suricatta) over a 6-10-week feeding trial to determine the acceptability of a commercially available manufactured diet intended for the feeding of insectivorous animals. Five animals at two zoos were heavier than desired and otherwise healthy at the start of the studies. Measurements of blood chemistries including cholesterol and cell blood count remained within physiologic expected ranges throughout the short-term study. Plasma and serum amino acid levels also remained within normal expected ranges, except taurine, which was much higher than predicted, although there were no significant changes in taurine over time. After initiation of the 6-week trial, these trials were extended for longer-term evaluation of animals in regards to body weight maintenance. In the long-term study, some individuals were restricted in their access to diet, and achieved a beneficial weight loss while maintaining good health. Approximate dietary consumption for captive meerkats in our study averaged 32-44 g, or calculated 92-127 kcal GE/meerkat*day (83-114 kcal ME/meerkat*day), and weight loss of animals at one institution of 10.4% was accomplished over 151 days from day 0, at approximately 30 g, or calculated 86 kcal GE/meerkat*day (78 kcal ME/meerkat*day). PMID- 19681146 TI - Enhancing captive breeding in giant pandas (Ailuropoda melanoleuca): maintaining lactation when cubs are rejected, and understanding variation in milk collection and associated factors. AB - From 1997 to 2002, a female giant panda (Ailuropoda melanoleuca) was artificially stimulated and lactation was maintained, after her neonates were removed due to the female's inability to provide maternal care. Milk samples were collected and the amount of milk collected was quantified. The lactation curve of this animal was estimated based on the Gamma function: Y(t)=at(b)e(-ct). The amount of milk collected showed significant, positive relationships with the number of days after parturition both in 1999 and in the whole study period from 1998 to 2002. This female's lactation curves fit the type I pattern of a typical mammalian lactation curve. Daily milk collection (g) during the first 30 days after parturition, and from 31 to 60 days after parturition, showed a consistent pattern with one peak at around 8:00 hr. More milk was collected during the latter period than during the former period. The amount of milk (g) collected on mucus excretion days was significantly less than that on days after mucus excretion had ended, yet no significant difference was found between milk collected one day before mucus days and on mucus days, or between milk collected one day before and one day after mucus days. Mucus excretion from the gastrointestinal tract significantly impacted the amount of milk collected. The results from this study may aid the captive propagation and conservation of giant pandas and other endangered and rare captive mammal species. PMID- 19681147 TI - Classifying ectodermal dysplasias: Incorporating the molecular basis and pathways (Workshop II). AB - Hereditary conditions are traditionally classified based either on physical/physiological attributes or using the names of the individuals credited with identifying the condition. For the 170 plus conditions classified as ectodermal dysplasias (EDs), both of these nosological systems are used, at times interchangeably. Over the past decade our knowledge of the human genome and the molecular basis of the EDs have greatly expanded providing the impetus to consider alternative classification systems. The incorporation of the molecular basis of hereditary conditions adds important information allowing effective transfer of objective genetic information that can be lacking from traditional classification systems. Molecular information can be added to the nosological system for the EDs through a hierarchical- and domain-based approach that encompasses the condition's name, mode of inheritance, molecular pathway affected, and specific molecular change. As new molecular information becomes available it can be effectively incorporated using this classification approach. Integrating molecular information into the ED classification system, while retaining well-recognized traditional syndrome names, facilitates communication at and between different groups of people including patients, families, health care providers, and researchers. PMID- 19681149 TI - Importance of PORCN and Wnt signaling pathways in embryogenesis. PMID- 19681150 TI - Captive reproduction of the Jamaican iguana (Cyclura collei). AB - The Jamaican iguana (Cyclura collei) was considered to be extinct until its rediscovery in 1990 in the Hellshire Hills region of Jamaica. Between 1994 and 1996, several wild caught juveniles were placed in six North American zoological institutions including the Indianapolis Zoo. The intent was to establish a self sustaining captive population outside of Jamaica in order to serve as an ancillary population should the wild numbers decline. Several environmental parameters such as temperature, humidity, photoperiod and diet were manipulated to encourage reproduction in captivity. In 2006, two clutches of eggs were deposited by two separate females for a total of 35 eggs. Twenty-six eggs were fertile and 22 hatched after a 76-83-day incubation at 30.3-30.8 degrees C (86.5 86.7 degrees F). The average weight of the neonates was 26.4 g and the average total length was 21.7 cm. This successful reproduction of C. collei constitutes the first North American hatching for this species. Environmental parameters, incubation techniques and neonate morphometry can serve as a baseline for further propagation of the species. This represents a positive milestone for the continuing conservation of this critically endangered species. PMID- 19681151 TI - Genetic testing in ectodermal dysplasia: availability, clinical utility, and the nuts and bolts of ordering a genetic test. AB - "Ectodermal Dysplasia syndromes" comprise a diverse group of heritable conditions characterized by congenital anomalies of one or more ectodermal structures and their appendages: hair, teeth, nails, and sweat glands. Genetic testing is available for many types of ectodermal dysplasia (ED) through clinical and/or research laboratories. We address the distinctions between genetic testing as performed on a clinical versus research basis, and summarize the clinical aspects, testing methodology, and sensitivity for those ED syndromes for which testing is available in a clinical laboratory. Lastly, we leave the laboratory for the clinical setting to discuss the utility of genetic testing for patients and their families, and summarize the practical issues involved in ordering a genetic test. PMID- 19681152 TI - 2008 International Conference on Ectodermal Dysplasias Classification: conference report. AB - There are many ways to classify ectodermal dysplasia syndromes. Clinicians in practice use a list of syndromes from which to choose a potential diagnosis, paging through a volume, such as Freire-Maia and Pinheiro's corpus, matching their patient's findings to listed syndromes. Medical researchers may want a list of syndromes that share one (monothetic system) or several (polythetic system) traits in order to focus research on a narrowly defined group. Special interest groups may want a list from which they can choose constituencies, and insurance companies and government agencies may want a list to determine for whom to provide (or deny) health-care coverage. Furthermore, various molecular biologists are now promoting classification systems based on gene mutation (e.g., TP63 associated syndromes) or common molecular pathways. The challenge will be to balance comprehensiveness within the classification with usability and accessibility so that the benefits truly serve the needs of researchers, health care providers, and ultimately the individuals and families directly affected by ectodermal dysplasias. It is also recognized that a new classification approach is an ongoing process and will require periodical reviews or updates. Whatever scheme is developed, however, will have far-reaching application for other groups of disorders for which classification is complicated by the number of interested parties and advances in diagnostic acumen. Consensus among interested parties is necessary for optimizing communication among the diverse groups whether it be for equitable distribution of funds, correctness of diagnosis and treatment, or focusing research efforts. PMID- 19681153 TI - Intake and digestion of horned guan Oreophasis derbianus diets measured in three Mexican zoos. AB - We conducted nutritional analyses of diets offered to and ingested by seven pairs of horned guans (Oreophasis derbianus) in three zoos. Digestibility was calculated with individually housed birds (n=1 at each zoo). Diets offered varied widely among institutions, both in ingredients fed as well as in nutrient composition. Feeding selectivity was evident through differences in composition of diets offered vs. consumed, with fruit (bananas, grape, and/or plantain) and avocado (when offered) highly preferred; green leaves, poultry pellets, and other vegetables comprised lesser proportions of the diet. All facilities fed 2-3X more food than consumed, allowing a great degree of choice of preferred items and potentially consumption of nutritionally imbalanced diets-in particular, mineral constituents. Diets were highly digestible; dry matter (DM) digestion coefficients ranged from 70 to approximately 90%; protein digestibility varied from 30 to 80%; fat was >90% digestible. Diet composition was compared with known nutritional requirements of domestic avian species, and feeding recommendations discussed. Despite the wide variability in nutrient composition of diets eaten (i.e. protein 6-10% of DM; fat 2-17% of DM), no overt health problems were noted and all pairs had successfully reproduced on these diets. It is suggested that horned guans may have nutrient requirements more similar to those suggested for other frugivorous birds than values determined for poultry as the physiologic model. Comparisons with native food items, as well as more detailed nutrient balance studies, may provide even better guidelines for captive management of this highly endangered species. PMID- 19681154 TI - Ectodermal dysplasias: clinical and molecular review. AB - Ectodermal dysplasias (EDs) as defined by Freire-Maia [Freire-Maia (1971); Hum Hered 21: 309-312; Freire-Maia (1977); Acta Genet Med Gemellol 26: 121-131] are congenital disorders characterized by alterations in two or more ectodermal structures, at least one of these involving alterations in hair, teeth, nails, or sweat glands. Suggestions for a new definition and, consequently, for a new classification of EDs have being proposed lately, mainly with the purpose of connecting clinical knowledge with recent discoveries of gene mutations responsible for about 30% of EDs. The aim of this review was to update the clinical classification of EDs with recent molecular (64 genes and 3 chromosome regions) and clinical data, mainly of EDs of the A group (N = 186), in order to contribute information for the evaluation of the ED definition proposed by Freire Maia. Our conclusion is that the combination of both procedures-clinical and molecular-only brings advantages for a deeper knowledge of EDs. First, it allows a rapid diagnosis that may become even more precise whenever DNA exams are available. Secondly, the comprehension of the biological mechanisms that cause EDs is needed for the design of efficient prevention and treatment approaches. PMID- 19681155 TI - New clinico-genetic classification of trichothiodystrophy. AB - Trichothiodystrophy (TTD) is a congenital hair dysplasia with autosomal recessive transmission. Cross banding pattern under polarized light plus trichoschisis and a low sulfur content of hair shafts define the disorder, which is associated with variable and neuroectodermal symptoms. So-called photosensitive forms of TTD (with low level of in vitro UV-induced DNA repair, not constantly associated with marked clinical photosensitivity) are caused by mutations in genes encoding subunits of the transcription/repair factor IIH (TFIIH). Ten percentage of nonphotosensitive patients are known to have TTDN1 mutations, the specific role of which is unknown. We studied nine patients recruited at our institution and reviewed 79 with molecular analysis out of 122 TTD patients reported in literature with the aim to collect systematically the clinical findings in TTD patients and establish genotype-phenotype correlations. The frequency of congenital ichthyosis, collodion-baby type, was significantly higher in the TFIIH mutated group. Hypogonadism was significantly more frequent in the non photosensitive group. There was no statistical difference regarding osseous anomalies. Mutations in TFIIH sub-units leading to abnormal expression in genes involved in epidermal differentiation could explain the particular dermatological changes seen in photosensitive cases of TTD. We suggest a new clinico-genetic classification of TTD, which may help clinicians confused by the current acronyms used (IBIDS, PIBIDS...). Understanding the TTD ichthyotic phenotype could lead to therapeutic advances in the management of TTD and other types of ichthyoses. PMID- 19681156 TI - Classifying functional manifestations of ectodermal dysplasias. AB - Ectodermal dysplasias (ED) encompass more than 200 conditions involving some combination of disorders of hair, nails, teeth, and sweat glands. The incidence of ED is relatively rare affecting about 7 of 10,000 births [Itin and Fistarol (2004)]. Individuals manifesting ED present with a wide range of disorders involving hair, nails, teeth, and sweat glands and in many cases other characteristics as well. The complex nature of the disorder has presented challenges for clinical practice and required the involvement of multiple approaches and disciplines. It has also resulted in significant research initiatives on cause and symptomatology. A significant challenge has been the search for comprehensive documentation of the varied and complex manifestations associated with ED. Existing classification systems of ED have focused on physiological and structural dimensions. Classification approaches with a broader focus including characteristics of functioning in persons with ED could facilitate clinical work and research initiatives. In this context, the potential utility of available classifications that address functioning and disability would be appropriate to consider in the search for a consensus classification of ED. To that end, the purpose of this article is to (a) review the status of classification of ED, (b) provide a brief overview of the International Classification of Functioning, Disability and Health-Children and Youth, ICF-CY [World Health Organization (2007); International Classification of Functioning, Disability and Health-Children and Youth. Geneva: WHO.], and (c) identify possible contributions of the ICF-CY to classification of ED's. PMID- 19681158 TI - Polarized expression of integrin beta1 in diencephalic roof plate during chick development, a possible receptor for SCO-spondin. AB - The roof plate of the caudal diencephalon is formed by the posterior commissure (PC) and the underlying secretory ependyma, the subcommissural organ (SCO). The SCO is composed by radial glial cells bearing processes that cross the PC and attach to the meningeal basement membrane. Since early development, the SCO synthesizes SCO-spondin, a glycoprotein that shares similarities to axonal guidance proteins. In vitro, SCO-spondin promotes neuritic outgrowth through a mechanism mediated by integrin beta1. However, the secretion of SCO-spondin toward the extracellular matrix that surrounds the PC axons and the expression of integrins throughout PC development have not been addressed. Here we provide immunohistochemical evidence to suggest that during chick development SCO cells secrete SCO-spondin through their basal domain, where it is deposited into the extracellular matrix in close contact with axons of the PC that express integrin beta1. Our results suggest that SCO-spondin has a role in the development of the PC through its interaction with integrin beta1. PMID- 19681157 TI - Discoidin domain receptor 2 (DDR2) is required for maintenance of spermatogenesis in male mice. AB - Discoidin domain receptor 2 (DDR2) is a receptor tyrosine kinase (RTK). We recently identified homozygous smallie mutant mice (BKS.HRS. Ddr2(slie/slie)/J, Ddr2(slie/slie) mutants), which lack a functional DDR2. Ddr2(slie/slie) mutant mice are dwarfed and infertile due to peripheral dysregulation of the endocrine system. To understand the role of DDR2 signaling in spermatogenesis, we studied the expression of several receptors, enzymes, and proteins related to spermatogenesis in wild-type and Ddr2(slie/slie) mutant mice at 10 weeks and 5 months of age. DDR2 were expressed in adult wild-type male mice in Leydig cells. The number of differentiated spermatozoa in the seminal fluid was significantly lower in the Ddr2(slie/slie) mutant mice than in the wild-type mice. The number of TUNEL-positive cells was significantly greater in 5-month-old Ddr2(slie/slie) mutants. Testosterone was significantly reduced at 5 months of age, but LH was similar in both types of mice at both 10 weeks and 5 months of age. The expression levels of LH receptors (Lhcgr), StAR, P450scc, and Hsd3beta6 were not significantly different between the two types of mice at 10 weeks of age, but they were significantly reduced in 5-month-old Ddr2(slie/slie) mutants compared to wild-type mice of the same age. DDR2 was expressed in the Leydig cells of adult wild-type male mice. In conclusion, our results indicated that DDR2 signaling plays a critical role in the maintenance of male spermatogenesis. PMID- 19681159 TI - Wnt signaling modulates pre- and postsynaptic maturation: therapeutic considerations. AB - Wnt signaling regulates a wealth of aspects of nervous system development and function in embryonic stages and in adulthood. The expression of Wnt ligands and components of the Wnt signaling machinery in early stages of neural development has been related to its role in neurite patterning and in synaptogenesis. Moreover, its expression in the mature nervous system suggests a role for this pathway in synaptic maintenance and function. Therefore, it is of crucial relevance the understanding of the mechanisms by which Wnt signaling regulates these processes. Herein, we discuss how different Wnt ligands, acting through different Wnt signaling pathways, operate in pre- and postsynaptic regions to modulate synapse structure and function. We also elaborate on the idea that Wnt signaling pathways are a target for the treatment of neurodegenerative diseases that affect synaptic integrity, such as Alzheimer's disease. PMID- 19681160 TI - Wnt won the war: antagonistic role of Wnt over Shh controls dorso-ventral patterning of the vertebrate neural tube. AB - The spinal cord has been used as a model to dissect the mechanisms that govern the patterning of tissues during animal development, since the principles that rule the dorso-ventral patterning of the neural tube are applicable to other systems. Signals that determine the dorso-ventral axis of the spinal cord include Sonic hedgehog (Shh), acting as a bona fide morphogenetic signal to determine ventral progenitor identities, and members of the Bmp and the Wnt families, acting in the dorsal neural tube. Although Wnts have been initially recognized as important in proliferation of neural progenitor cells, their role in the dorso ventral patterning has been controversial. In this review, we discuss recent reports that show an important contribution of the Wnt canonical pathway in dorso ventral pattern formation. These data allow building a model by which the ventralizing activity of Shh is antagonized by Wnt activity through the expression of Gli3, a potent inhibitor of the Shh pathway. Therefore, antagonistic interactions between canonical Wnt, promoting dorsal identities, and Shh pathways, inducing ventral ones, would define the dorso-ventral patterning of the developing central nervous system. PMID- 19681162 TI - C-terminal deletion of the atrophin-1 protein results in growth retardation but not neurodegeneration in mice. AB - Dentatorubral-pallidoluysian atrophy (DRPLA) is a dominant hereditary neurodegenerative disorder caused by the expansion of a poly-glutamine (poly-Q) repeat in Atrophin-1 protein. Ectopic expression of a poly-Q expanded human Atrophin-1 is sufficient to induce DRPLA phenotypes in mice. However, it is still unclear whether the dominant effect of poly-Q expansion is due to the functional interference with wild-type Atrophin-1 proteins, which exist in both patients and transgenic mice. Here we report the generation and analysis of an Atrophin-1 targeting allele that expresses a truncated protein lacking both the poly-Q repeat and following C-terminal peptides. Homozygous mutants exhibit growth retardation and progressive male infertility, but no obvious signs of neurodegeneration. Moreover, the mutant allele neither blocked nor enhanced the neurodegenerative phenotypes caused by a poly-Q expanded transgene. These results support the model that poly-Q expanded Atrophin-1 proteins cause DRPLA in a manner independent of any functional interaction with wild-type Atrophin-1 proteins. PMID- 19681161 TI - Differential expression patterns and developmental roles of duplicated scinderin like genes in zebrafish. AB - Scinderin, the closest homologue of the actin-severing protein, gelsolin, has two similar paralogs (Scinla and Scinlb) in zebrafish. Scinla is abundant in the adult cornea; Scinlb comprises considerably less corneal protein. Here, we show that scinla is expressed in the nose, lens, brain, cornea and annular ligament of the iridocorneal angle; by contrast, scinlb is expressed in the hatching gland, floor plate, notochord, otic vesicle, brain, pharynx, cartilage, swim bladder and cornea. Activity of scinla and scinlb promoter fragments driving the EGFP reporter gene in transgenic zebrafish resembled scinla or scinlb expression. Previously, we showed that reduction of scinla by injection of antisense morpholino oligonucleotides ventralized embryos; here, specific reduction of scinlb expression led to subtle brain abnormalities associated with increased cell death, decreased shhb expression in the floor plate, and slightly reduced eye distance. Thus, scinla and scinlb have different expression patterns and developmental roles during zebrafish development. PMID- 19681163 TI - The Pitx2c N-terminal domain is a critical interaction domain required for asymmetric morphogenesis. AB - The paired-like homeodomain transcription factor Pitx2c has an essential role in patterning the left-right axis. However, neither its transcriptional targets nor the molecular mechanisms through which it exerts its patterning function are known. Here we provide evidence that the N-terminal domain of Pitx2c is important for this activity. Overexpression of the Pitx2c N-terminus in ovo randomizes the direction of heart looping, the first morphological asymmetry conserved in vertebrate embryos. In addition, the Pitx2c N-terminal domain blocks the ability of Pitx2c to synergize with Nkx2.5 to transactivate the procollagen lysyl hydroxylase (Plod-1) promoter in transient transfection assays. A five amino acid region containing leucine-41 is required for both of these effects. Our data suggest that the Pitx2c N-terminal domain competes with endogenous Pitx2c for binding to a protein interaction partner that is required for the activation of genes that direct asymmetric morphogenesis along the left-right axis. PMID- 19681165 TI - Single-needle laser treatment with drainage of hydrothorax in fetal bronchopulmonary sequestration with hydrops. AB - Bronchopulmonary sequestration (BPS) is sometimes associated with hydrothorax and hydrops in utero. In the absence of fetal hydrops, perinatal outcome is favorable and justifies expectant management. In the presence of fetal hydrops, perinatal outcome is reported to be extremely poor and intervention should be considered. Therapeutic options include open fetal surgery, minimally invasive coagulation of the blood supply and thoracoamniotic shunting. We present the first case of fetal hydrops and a large hydrothorax due to BPS treated successfully with one ultrasound-guided thin needle insertion, through which both laser coagulation of the feeding artery and drainage of the hydrothorax were performed. Following the procedure the hydrothorax and hydrops gradually disappeared and the BPS diminished in size. A healthy neonate was delivered uneventfully at term. We describe the case and discuss the different therapeutic options. PMID- 19681166 TI - Reticulon 3 is an interacting partner of the SALM family of adhesion molecules. AB - Synaptic adhesion-like molecules (SALMs) are a recently discovered family of adhesion molecules that is widely distributed in the central nervous system and has been implicated in neurite outgrowth and synapse formation. To identify proteins that interact with extracellular domains of SALMs, we carried out yeast two-hybrid screening using the extracellular domain of SALM1 as bait. A clone encoding full-length reticulon 3A1 was isolated. This interaction was shown to occur through the LRR domain, which is found on all SALMs. To determine whether this relationship also occurs in brain, we performed immunoprecipitation using antibodies to SALMs 1-4. A 19-kDa band, identified as reticulon 3C, bound to all four SALMs, whereas a 90-kDa band, which did not comigrate with any known reticulon 3 variant, bound to SALMs 2 and 3. These results show that reticulon 3 may play a role in the trafficking of the SALM family of adhesion molecules. PMID- 19681167 TI - Single doublecortin gene therapy significantly reduces glioma tumor volume. AB - We employed lentivirus-based doublecortin (DCX), as a glioma suppressor gene therapy in an intracranial glioma tumor xenograft model in nude rats. Single DCX expressing lentivirus was directly administered into the tumor on day 8 after U87 tumor cell implantation. DCX treatment significantly reduced U87 glioma tumor volume (approximately 60%) on day 14 after DCX lentivirus injection and significantly improved median survival of tumor-bearing nude rats. DCX synthesis induced neuronal markers MAP2, TUJ1, and PSA-NCAM and the glial marker glial fibrillary acidic protein (GFAP) in the implanted U87 glioma tumors. DCX synthesis induced GFAP that colocalized with tubulin in the mitotic stage, inhibited cleavage furrow during cytokinesis, and blocked mitosis in glioma cells. DCX lentivirus infection did not induce apoptosis but significantly inhibited expression of the proliferation marker Ki-67 and the blood vessel marker von-Willebrand factor (vWF). U87 and other glioma cells except for brain tumor stem cells (BTSCs) do not express neuronal markers or both neuronal and glial markers. DCX-synthesizing glioma cells express a phenotype of antiangiogenic BTSC-like cells with terminal differentiation that causes remission of glioma cells by blocking mitosis via a novel DCX/GFAP pathway. Direct local delivery of lentivirus-based DCX gene therapy is a potential differentiation-based therapeutic approach for the treatment of glioma. PMID- 19681168 TI - Somatic Ca2+ signaling in cerebellar Purkinje neurons. AB - Activity-driven Ca(2+) signaling plays an important role in a number of neuronal functions, including neuronal growth, differentiation, and plasticity. Both cytosolic and nuclear Ca(2+) has been implicated in these functions. In the current study, we investigated membrane-to-nucleus Ca(2+) signaling in cerebellar Purkinje neurons in culture to gain insight into the pathways and mechanisms that can initiate nuclear Ca(2+) signaling in this neuronal type. Purkinje neurons are known to express an abundance of Ca(2+) signaling molecules such as voltage-gated Ca(2+) channels, ryanodine receptors, and IP3 receptors. Results show that membrane depolarization evoked by brief stimulation with K(+) saline elicits a prominent Ca(2+) signal in the cytosol and nucleus of the Purkinje neurons. Ca(2+) influx through P/Q- and L-type voltage-gated Ca(2+) channels and Ca(2+) induced Ca(2+) release (CICR) from intracellular stores contributed to the Ca(2+) signal, which spread from the plasma membrane to the nucleus. At strong K(+) stimulations, the amplitude of the nuclear Ca(2+) signal exceeded that of the cytosolic Ca(2+) signal, suggesting the involvement of a nuclear amplification mechanism and/or differences in Ca(2+) buffering in these two cellular compartments. An enhanced nuclear Ca(2+) signal was more prominent for Ca(2+) signals elicited by membrane depolarization than for Ca(2+) signals elicited by activation of the metabotropic glutamate receptor pathway (mGluR1), which is linked to Ca(2+) release from intracellular stores controlled by the IP3 receptor. PMID- 19681170 TI - Live FISH: imaging mRNA in living neurons. AB - This paper describes a novel technique to identify and study neurons that express specific mRNAs. The method is called Live FISH because it is like fluorescent in situ hybridization (FISH) but can be applied to living tissues. In Live FISH, molecular beacons, which fluoresce only in the presence of the complementary mRNA, are delivered by gene gun into living neurons. One of its many benefits over existing gene expression assays is that the neurons are alive. Because identified neurons are living, they can be targeted for subsequent single-cell assays, such as electrophysiology. In addition, dye delivered with the molecular beacons illuminates dendritic morphology. Live FISH is cheaper than generating transgenic animals, and the endogenous mRNA is assayed without covalent modification or genetic manipulation. This report demonstrates the feasibility of Live FISH with yellow fluorescent protein (YFP) mRNA in living mouse retinas, although molecular beacons can be rationally designed to any mRNA of interest. PMID- 19681169 TI - Melatonin treatment potentiates neurodegeneration in a rat rotenone Parkinson's disease model. AB - Parkinson's disease (PD) is characterized pathologically by progressive neurodegeneration of the nigrostriatal dopamine (DA) system. Currently, the cause of the disease is unknown, except for a small percentage of familial cases (<10% of total). The rat rotenone model reproduces many of the pathological features of the human disease, including apomorphine-responsive behavioral deficits, DA depletion, loss of striatal DA terminals and nigral dopaminergic neurons, and alpha-synuclein/polyubiquitin-positive cytoplasmic inclusions reminiscent of Lewy bodies. Therefore, this model is well-suited to examine potential neuroprotective agents. Melatonin is produced mainly by the pineal gland and is known primarily for regulating circadian rhythms. It also has potent free radical scavenging and antiinflammatory properties. Melatonin has been reported to be neuroprotective in the 6-hydroxydopamine (6-OHDA) and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) models of PD. However, there are conflicting reports suggesting that melatonin does not provide neuroprotection in these models. Melatonin elicits significant functional changes in the nigrostriatal DA system that may affect 6 OHDA and MPTP entry into cells. Therefore, rotenone is an ideal model for assessing protection, because it does not rely on the dopamine transporter uptake to exert neurotoxicity. In this study, the neuroprotective potential of melatonin in the rotenone PD model was assessed. Melatonin potentiated striatal catecholamine depletion, striatal terminal loss, and nigral DA cell loss. Indeed, melatonin alone elicited alterations in striatal catecholamine content. Our findings indicate that melatonin is not neuroprotective in the rotenone model of PD and may exacerbate neurodegeneration. PMID- 19681164 TI - Wnt to build a tube: contributions of Wnt signaling to epithelial tubulogenesis. AB - Epithelial tubes are crucial to the function of organ systems including the cardiovascular system, pulmonary system, gastrointestinal tract, reproductive organ systems, excretory system, and auditory system. Using a variety of animal model systems, recent studies have substantiated the role of Wnt signaling via the canonical/beta-catenin-mediated trajectory, the non-canonical Wnt trajectories, or both, in forming epithelial tubular tissues. This review focuses on the involvement of the Wnt pathways in the induction, specification, proliferation, and morphogenesis involved in tubulogenesis within tissues including the lungs, kidneys, ears, mammary glands, gut, and heart. The ultimate goal is to describe the developmental processes forming the various tubulogenic organ systems to determine the relationships between these processes. PMID- 19681171 TI - Regulation of activin A synthesis in microglial cells: pathophysiological implications for bacterial meningitis. AB - Previous studies have shown that activin A, a neuroprotective cytokine and dimeric polypeptide composed of two betaA subunits, is elevated in the cerebrospinal fluid of patients suffering from bacterial meningitis. In this study, to elucidate further the functional significance and pathophysiological implications of these findings, we demonstrated that microglial cells are not only the source but also the target cells of activin A in the central nervous system: immunohistochemistry and RT-PCR revealed expression of activin subunit betaA mRNA as well as activin receptor type I and type II mRNA in rat microglia in vitro. Further studies showed that activin enhances microglial proliferation and decreases the gamma-interferon-induced synthesis of nitric oxide, one of several microglial mediators involved in the inflammatory response in microglia activation. Furthermore, quantitative RT-PCR, Western blotting, and ELISA showed an inhibitory effect of activin on inducible nitric oxide synthase, tumor necrosis factor-alpha, interleukin-6, and interleukin-1beta gene and protein levels after lipopolysaccharide treatment. We suggest that the increased synthesis of activin A is directly involved, via influence on microglia cell functions, in the modulation of the inflammatory response in bacterial meningitis. PMID- 19681172 TI - Differentiation of dementia with Lewy bodies from Alzheimer disease using the frontal assessment battery test. PMID- 19681173 TI - Out-of-hospital medication errors: a 6-year analysis of the national poison data system. AB - PURPOSE: Previous research and reporting has focused on the incidence and prevention of medication errors in the hospital setting; however, no previous study has reported the frequencies, characteristics and outcomes of out-of hospital medication (OHME) errors. METHOD: Data from the National Poison Data System (NPDS) was collected for 2000-2005 and information regarding out-of hospital medication errors reported to Poison Control Centers (PCC) was collected by a trained investigator. RESULTS: From 2000-2005 there were 1,166,116 OHME reported to PCC. Of these patients, 88,451 (7.5%) received medical evaluation by a healthcare provided and 229 (0.01%) deaths reported. The most common drug classes involved included cough/cold medications, analgesics, cardiovascular agents, antihistamines, antidepressants and antimicrobial agents. The most common error reported in both children and adults was taking or giving medication twice. CONCLUSIONS: OHME occur frequently and the NPDS may be a useful resource for data collection and evaluation in this previously overlooked population. The majority of OHME reported did not result in any significant morbidity or mortality and were managed at home without need for healthcare referrral. Further study of OHME is needed, and in particular whether healthcare professionals can target educational instruction to patients so as to effectively reduce the frequency of the most common or injurious errors. PMID- 19681175 TI - [Does one have a right to death without resuscitation attempts?]. PMID- 19681174 TI - Characterization of chromosomal integration sites for heterologous gene expression in Saccharomyces cerevisiae. AB - The construction of mitotically stable yeast strains for heterologous gene or pathway expression often requires chromosomal integration. However, transcription levels vary between different chromosome regions. We therefore characterized 20 different integration sites of the Sacchromyces cerevisiae genome by inserting lacZ as a reporter gene under the control of two different promoters and determining expression levels through enzyme activity measurement. An up to 8.7 fold difference was detected between the sites conferring lowest and highest expression, respectively. This opens the opportunity for modulating gene expression levels while retaining promoter and culture conditions. PMID- 19681177 TI - Vaccine refusal and the risks of vaccine-preventable diseases. PMID- 19681176 TI - Dipyridamole plus aspirin and hemodialysis graft patency. PMID- 19681178 TI - Case 15-2009: a man with coma after cardiac arrest. PMID- 19681179 TI - Case 15-2009: a man with coma after cardiac arrest. PMID- 19681180 TI - [Topical skin care in nursing homes]. PMID- 19681181 TI - Bolivia tackles maternal and child deaths. PMID- 19681182 TI - Proceedings and abstracts of the 8th International Veterinary Immunology Symposium, August 15-19, 2007, Ouro Preto, Brazil. PMID- 19681184 TI - Model evacuation plan. PMID- 19681185 TI - Not so silent nurse. PMID- 19681186 TI - Holmesglen debate continues. PMID- 19681188 TI - E-health e-volves. PMID- 19681189 TI - South Africa tries new approach to resistant tuberculosis. PMID- 19681190 TI - Reality check for checklists. PMID- 19681191 TI - Cognitive behavior therapy may ease anxiety. Changing the way you think about problems can reduce worry and lead to healthier behaviors. PMID- 19681192 TI - I've heard that high-fat foods and alcohol can raise triglyceride levels. I take Lipitor for high cholesterol, don't eat high-fat foods and don't drink alcohol except for half a glass of wine a few times a month. My triglycerides are always high when checked--my last reading was 222. What else can I do to help lower my triglycerides? PMID- 19681193 TI - Clash of the titans. The pacemaker and the MRI can't work together--at least not yet. PMID- 19681194 TI - The riddle of MyPyramid. The new food pyramid makes a mystery of healthful eating. PMID- 19681195 TI - Finding your way through the (mini) Maze. Minimally invasive operations for atrial fibrillation are on the rise. How well they work is up in the air. PMID- 19681196 TI - Bone thinning linked to blood thinner. PMID- 19681198 TI - Radiation in medicine: a double-edged sword. X-rays, CT scans, and other procedures should be used judiciously. PMID- 19681197 TI - Ask the doctor. My doctor and I are confused by my cholesterol levels. My good cholesterol is extraordinarily high, at 94, but my bad cholesterol is on the high side, too, at 154. To resolve this good news/bad news story, my doctor ordered a C-reactive protein test which, of course, came back right in the middle of the normal range. I don't smoke or have other factors for heart disease. Would you advise me at start a statin or some other drug? PMID- 19681199 TI - Women's hearts need extra attention. Diagnosis and treatment of heart disease in women still lags behind that in men. PMID- 19681200 TI - Potassium and sodium out of balance. Too little potassium and too much sodium is bad for the heart and general health. PMID- 19681201 TI - Binge drinking and stroke. PMID- 19681202 TI - Ask the doctor. My husband is taking arginine and citrulline supplements because he read that they will protect his heart and arteries. Should I try these supplements, too, or is this a waste of money? PMID- 19681203 TI - Ask the doctor. I have an electrocardiogram as part of my yearly checkup. After the last one, my doctor mentioned that my ST segment was longer this year than it was last year. He recommended that I have a stress test to check this out. I passed with flying colors. When I asked the cardiologist who did the stress test about the ST segment, he said the length isn't really important, that the height and shape are what matter. Can you explain? PMID- 19681204 TI - Defining a moderate-intensity workout. A simple walking test can gauge the intensity of your walking pace. PMID- 19681205 TI - Exercise equals angioplasty for leg pain. The slow-but-steady approach has benefits the quick fix can't offer. PMID- 19681206 TI - Treat "mini-strokes" as an emergency, not a gentle warning. Evaluating a transient ischemic attack right away can prevent a stroke. PMID- 19681207 TI - New guidelines refine aspirin prescription. Task force offers recommendation for women and guidance on dosage. PMID- 19681208 TI - Atrial fibrillation and blood pressure. PMID- 19681209 TI - Ask the doctor. After being diagnosed with high blood pressure several years ago, I started taking diltiazem and Atacand. The results were good, giving me an average blood pressure of 110/65. I recently developed gastroenteritis and aspiration pneumonia. While I was in the hospital, my blood pressure got so low I was told to stop taking these medications. I have been off them since, and my blood pressure has remained normal, averaging 105/65. How can this be? Will high blood pressure return? PMID- 19681210 TI - Ask the doctor. I am due to have a colonoscopy. My cardiologist told me that I will need to stop taking Coumadin, which I take for atrial fibrillation, a few days before the procedure and get some injections. Is this really necessary? PMID- 19681211 TI - [Suicide patterns and characteristics in Akita, Japan]. PMID- 19681212 TI - [Psychological effects brought about by the exposure to the atomic bomb]. PMID- 19681214 TI - [Psychological effects and intervention following a traffic disaster involving a large number of victims]. PMID- 19681213 TI - [Morbidity survey on community-residing aged individuals 3 years after Niidata Chuetsu Earthquake]. PMID- 19681215 TI - Physical anthropology: the search for general processes and principles. AB - Physical anthropology consists of two interdependent types of study: (1) the biological history of man and (2) general biological processes in man (such as mechanisms of evolution and growth). Popular interest may focus on the former, the fascinating story of the origin of man and of specific people, but the latter affords physical anthropology potential practical value in respect to medicine, dentistry, public health, and population policy. The study of general processes is the study of human beings in particular situations, not for what we can learn about these particular populations but for the sake of generalization about mankind anywhere in comparable situations. This is, of course, the purpose of experimental science in general, but in anthropology the method is usually comparative. Long ago the study of the growth of the two sexes and of children in different countries was started on a comparative basis as was the study of the so called secular change in adult stature. By 1911 Franz Boas had compared the changes in stature and head form of children of several different immigrant groups in the United States. There have since been comparative studies of the amount and distribution of body fat (but not yet adequate comparative measurements of the relation of tissue components to diet and to diseases). Demographic patterns, inbreeding, outbreeding, and their effects are other general problems. The Human Adaptability Project of the International Biological Program promises studies of human response to heat, cold, altitude, and other conditions on a wide international basis. If supported, these could turn physical anthropology's search in a useful direction. The functional biology of people of even out-of-the-way communities will be compared with each other. These studies can yield general statements concerning human response to types of ecological situation including such sociocultural conditions as those of hunting-gathering tribes and urban slums. PMID- 19681216 TI - The psychological interdependence of family, school, and bureaucracy in Japan. AB - The Japanese "examination hell" phenomenon is viewed as a series of crisis rites through which the child passes from family-centered to peer group - centered values in a "particularistic" society. It is held that this model has greater explanatory power than the "minimization of competition" model proposed by others and that it also helps to explain the phenomenon of student radicalism and centrifugal relationships in middle-class communities. PMID- 19681217 TI - On Mexican folk medicine. AB - In this paper traditional medical beliefs and practices in a Mexican village are described and interpreted. The analysis focuses on the notion that health is a balance of hot and cold within the body. Several lines of evidence are used to reveal the metaphorical meanings of hot and cold, and these meanings are then seen to be related to structural features of peasant society. PMID- 19681218 TI - Vindolanda and the dating of Roman footwear. PMID- 19681219 TI - Review: ritual and interpretation in Provincial Roman cemeteries. PMID- 19681220 TI - A Roman bath-house at Duntocher on the Antonine Wall. PMID- 19681221 TI - Animal remains from temples in Roman Britain. PMID- 19681222 TI - "Wandering about" and depression: more examples. PMID- 19681223 TI - "Wandering about" as a topos of depression in Ancient Near Eastern literature and in the Bible. PMID- 19681224 TI - "When you perform the ritual of 'rubbing'": on medicine and magic in Ancient Mesopotamia. PMID- 19681225 TI - Thinking with drinking: wine and the Symposium in Aristophanes. PMID- 19681226 TI - [Poverty in Turkish cinema and the poor heroes]. PMID- 19681227 TI - [Territorial and agrarian rights: land mobility and land market after 18 years of neoliberal politics in Mexico]. PMID- 19681228 TI - [Poverty and richness in novels]. PMID- 19681229 TI - [The power of the rural!]. PMID- 19681231 TI - Socratic suicide. AB - When is it rational to commit suicide? More specifically, when is it rational for a Platonist to commit suicide, and more worryingly, is it ever not rational for a Platonist to commit suicide? If the Phaedo wants us to lear that the soul is immortal, and that philosophy is a preparation for a state better than incarnation, then why does it begin with a discussion defending the prohibition of suicide? In the course of that discussion, Socrates offers (but does not necessarily endorse) two arguments for the prohibition of self-killing, at least in most circumstances, which have exerted a long and powerful influence over subsequent discussion of the topic, particularly in theist contexts. In the context of the Phaedo itself, however, this introductory conversation plays a crucial role in setting the agenda for the remainder of the dialogue and offering an initial discussion of the major concerns of the argument as a whole. In par- ticular, the discussion of thte nature of suicide is intimately bound up with Socrates' conception of true philosophy as a 'preparation for death', the relationship between soul and body, and the immortality of the soul. My intention is to provide a reading of that passage (61e-69e) which asks whether the Phaedo can offer a philosophically satisfying distinction between suicide and philosophy and how it relates to other ancient philosophical attitudes to self-killing. I argue that Socrates does not think that being dead is always preferable to being alive, and that the religious views expressed in the passage are consistent with his general stance on the benevolence of the gods. PMID- 19681230 TI - [The restructuring and internationalization of the food sector in Turkey]. PMID- 19681232 TI - Cardiac computed tomography: current practice and future applications. AB - Cardiac computed tomography (CT) has continued to make great strikes in cardiovascular imaging. New advances and refinements in technology have enable CT to render astonishingly detailed images. Cardiac CT depends on a high temporal resolutions to minimize coronary artery movement-related motion artifacts. There is ongoing debate as to the position of CT in the algorithm of diagnostic imaging in the realm of the cardiovascular sciences. There is a definite role for cardiac CT in patients with chest pain who have low or intermediate probability of coronary artery disease; a negative result on cardiac CT angiography could potentially exclude significant coronary artery disease and thus obviate further expensive workup. Further advancements in technique and validation of results will help establish CT on firmer ground. Such developments should proceed in tandem with reimbursement practices that encourage use of this tool in the right clinical context. PMID- 19681233 TI - The Greek demographic expansion: models and comparisons. AB - For much of the first millennium BC, the number of Greeks increased considerably, both in the Aegean core and in the expanding periphery the Mediterranean and Black Sea regions. This paper is the first attempt to establish a coherent quantitative framework for the study of this process. In the first section, I argue that despite the lack of statistical data, it is possible to identify a plausible range of estimates of average long-term demographic growth rates in mainland Greece from the Early Iron Age to the Classical period. Elaborating on this finding, the second section offers a comprehensive rebuttal of the notion of explosive population growth in parts of the eighth and seventh centuries BC. In the third section, I seek to determine the probable scale and demographic consequences of Greek settlement overseas. A brief preliminary look at the relationship between population growth and the quality of life concludes my survey. The resultant series of interlocking parametric models is meant to contextualize the demographic development of ancient Greece within the wider ambit of pre-modem demography, and to provide a conceptual template for future research in this area PMID- 19681234 TI - Familiarity breeds: incest and the Ptolemaic Dynasty. AB - This paper examines the problem of Ptolemaic incest from a variety of cross disciplinary perspectives. Specifically, it seeks to establish the following: that there is little in the ancient record to support the common claim that the Ptolemies suffered extensively from the deleterious genetic effects of inbreeding; that the various theories so far put forward as explanations for Ptolemaic incest offer at best only a partial rationale for this dynastic practice; that the most compelling rationale for Ptolemaic incest is to be found in complex, and perhaps unconscious, symbolic motivations analogous to those observed by anthropologists in other cultures; and finally, that, for the Ptolemies, incest was, like the "truphe" for which they were so notorious, a dynastic signature which highlighted their singularity and above all, their power. PMID- 19681235 TI - Introduction. PMID- 19681236 TI - The role of LTRAs in the management of persistent asthma. AB - Two leukotriene receptor antagonists (LTRAs)--montelukast and zafirlukast--are currently available in the United States. Montelukast is approved for the treatment of asthma in patients as young as 2 years old and zafirlukast, in patients 7 years of age or older. In more than 15 clinical trials of LTRA therapy for mild to moderate persistent asthma, both of these oral agents produced rapid improvement in pulmonary function and daytime asthma symptoms. They also decreased the frequency of nocturnal awakening and the need for rescue therapy with beta2-adrenergic agonists and oral corticosteroids. These effects were maintained throughout the treatment period; tolerance did not develop. When used concomitantly with an inhaled corticosteroid (ICS), each LTRA further improved asthma control, thus permitting the partial or complete tapering off of the ICS dose needed to maintain clinical control. Although rare, previously undiagnosed cases of Churg-Strauss syndrome have been unmasked when ICS use is decreased or discontinued. The improved adherence gained with an oral agent administered as infrequently as once daily maximizes the effectiveness of these newest antiasthma medications. PMID- 19681237 TI - Dialysis in the Kingdom of Saudi Arabia. PMID- 19681238 TI - [Study on psychiatric disorders that develop one month after injuries associated with traffic accidents and factors to predict the development of such disorders]. PMID- 19681239 TI - [Mental health of the families of crime victims and factors related to their recovery]. PMID- 19681240 TI - [Post-partum depression and support extended to child care]. PMID- 19681241 TI - A picture's worth. PMID- 19681242 TI - The team approach. PMID- 19681243 TI - Prof Dr Peter Scharer. May 17, 1933. December 22, 2004. PMID- 19681244 TI - Peter Scharer--a photo essay. PMID- 19681245 TI - [Depression among housewives and couple therapy]. PMID- 19681246 TI - [Psychiatric aspect of brain trauma]. PMID- 19681247 TI - [PDFTBI and temporal pole lesion--hypothetical cognitive mechanism of delusional perception]. PMID- 19681248 TI - [Basic information necessary for diagnosis of epilepsy by psychiatrists]. PMID- 19681249 TI - [One hundred years of Seishin Shinkeigaku Zasshi--1909. Dr. Shuzo Kure: The relationship between beri-beri and psychiatric disease]. PMID- 19681250 TI - You're the flight surgeon: a psychiatric patient in a combat zone. PMID- 19681251 TI - Tooth whitening clinical trials: a global perspective. AB - Tooth whitening has been the subject of extensive clinical trials research since the introduction of the first hydrogen-peroxide whitening strips in 2000. Availability of digital image analysis, an unambiguous and reproducible method for assessing color change, has contributed to global clinical research and product development on whitening strips. The research has included a series of global randomized controlled trials in distinct sites and cultures, involving 6 6.5% hydrogen peroxide whitening strips used for 7-21 days. These studies, conducted at research hospitals, dental schools, and private dental practice, demonstrated significant color improvement with whitening strips relative to baseline and/or various controls without serious adverse events. This integrated clinical trials research provides important evidence of long-term safety and effectiveness of tooth whitening with 6-6.5% hydrogen peroxide whitening strips. PMID- 19681252 TI - Application of digital imaging in tooth whitening randomized controlled trials. AB - PURPOSE: The development of novel peroxide-based bleaching systems during the last several years has prompted the need for robust clinical methods to evaluate whitening response. Advances in digital camera technology and image analysis software provided the basis for an instrumental method to assess tooth color closely following a technique previously used to quantify plaque on tooth surfaces. In vitro and in vivo research was conducted to determine reproducibility of color measurements using this objective, digital imaging method. METHODS: Each of the 16 tabs in a standard shade guide system was mounted in a jig, and measurement reproducibility was assessed in vitro from paired digital images collected over a 2-day period. Separately, clinical measurement reproducibility was assessed in vivo from paired images of 14 healthy adult volunteers collected over a 2-day period. From these digital images, mean L*, a*, and b* color values were derived for each of the 16 individual shade tabs (in vitro study), or the facial surfaces of the maxillary six anterior teeth (in vivo study) of the 14 subjects. For each data set, variability was determined using ANOVA, and between-visit color measurement reliability was determined from intra class correlation coefficients (ICCs). RESULTS: In the in vitro study, shade tab yellowness (b*) ranged from 9.0-18.6, lightness (L*) ranged from 63.4-76.2, and redness (a*) ranged from 0.9-3.6. Overall daily means differed by 0.08 units or less, and intra-class correlations for the image pairs were 0.998 for L*, 0.996 for a* and 0.998 for b*. In the in vivo assessment, the 14 volunteers exhibited considerable range in tooth color. Yellowness (b*) ranged from 13.5-21.3, lightness (L*) ranged from 69.2-78.0, and redness (a*) ranged from 5.2-8.8. Clinical measurement of mean tooth color from digital images was highly reproducible across visits. Intra-class correlations for the image pairs were 0.989 for b*, 0.970 for L* and 0.979 for a*. PMID- 19681253 TI - Professional whitening strips in a university population. AB - PURPOSE: To evaluate the clinical response of a professional whitening strip system used by a university-based population residing in Mexico City, Mexico. METHODS: A randomized, double-blind, placebo-controlled study was conducted to evaluate the safety and efficacy of 6.5% hydrogen peroxide whitening strips used over a 3-week period. A total of 30 volunteer students and staff at the National Autonomous University of Mexico (Mexico City) were randomly assigned to the peroxide or placebo strip groups. Strips were worn for 30 minutes two times a day for 3 weeks. Efficacy was evaluated using digital image analysis to assess change in L* a* b* tooth color, while safety was assessed by oral examination and subject interview. RESULTS: Relative to placebo, the 6.5% hydrogen peroxide strip group experienced nearly a 4-unit color improvement (Deltab*). Treatment groups differed significantly (P< 0.0001) with respect to yellowness (Deltab*), lightness (DeltaL*) and redness (Deltaa*). Adjusted mean (SE) overall color improvement (DeltaW*) was -4.76 (0.27) for the peroxide strips, compared to the near zero, -0.21 (0.28) for the placebo control. Strip use was well tolerated. Minor, transient tooth sensitivity occurred more frequently in the peroxide group, and overall, no subjects modified or discontinued treatment early because of adverse events. PMID- 19681254 TI - Daytime use of a custom bleaching tray or whitening strips: initial and sustained color improvement. AB - PURPOSE: To compare the clinical response of 6% hydrogen peroxide whitening strips and a 10% carbamide peroxide custom tray system under common daytime usage conditions, in an Italian dental research center. METHODS: Informed consent and baseline measurements were collected, and 43 healthy adults were randomly assigned to 6% hydrogen peroxide whitening strips (Crest Whitestrips) or the 10% carbamide peroxide custom tray (Opalescence 10%). The maxillary arch was treated twice daily for 30 minutes at-home. Treatment was discontinued after 2 weeks, and subjects were monitored for an additional 4 weeks. Efficacy (initial and sustained) was measured objectively from standard digital images of the maxillary facial tooth surfaces using the international CIELAB system. Safety was assessed from interview and examination. Treatments were compared after 2 weeks (end-of treatment) and 6 weeks (4 weeks post-treatment) using analysis of covariance methods. RESULTS: Both groups exhibited color improvement at the Week 2 end-of treatment visit. For yellowness, mean (SD) Deltab* at Week 2 was -2.10 (0.70) for the strip group and -1.61 (1.03) for the tray group. For lightness, mean (SD) DeltaL* at Week 2 was 1.25 (0.92) for the strip group and 1.17 (1.19) for the tray group. Compared to Week 2, the strip group retained 89-92% of the initial Deltab* and DeltaL* color improvement at Week 6 (4 weeks post-treatment), while the tray group had 80-90%. Groups differed significantly (P< 0.05) on end-of treatment and post-treatment Deltab*, favoring the strips. Both daytime treatments were well-tolerated, with minor tooth sensitivity and oral irritation representing the most common findings. PMID- 19681255 TI - Clinical trial of long-term color stability of hydrogen peroxide strips and sodium percarbonate film. AB - PURPOSE: To compare initial and sustained clinical response of 6% hydrogen peroxide whitening strips and a 19% sodium percarbonate film in a randomized controlled trial. METHODS: Informed consent was obtained, after which 72 subjects were randomized to 6% hydrogen peroxide whitening strips (Crest Whitestrips), 19% sodium percarbonate brush-applied gel that dries as a film (Crest Night Effects), or placebo brush-applied gel without peroxide. Efficacy (digital imaging) and safety (clinical examination and interview) were assessed after 2 weeks treatment, and again at up to eight post-treatment timepoints over an 18-month post-treatment period. RESULTS: For Deltab* (yellowness), end-of-treatment adjusted means +/- standard errors (SE) were -2.37 +/- 0.088 for the strip group, -1.36 +/- 0.091 for the film group, and -0.08 +/- 0.090 for the placebo group. For DeltaL* (brightness), end-of-treatment adjusted means +/- SE were 2.40 +/- 0.121 for the strip group, 1.47 +/- 0.125 for the film group, and 0.06 +/- 0.122 for the placebo group. Groups differed significantly (P< 0.02) at end-of treatment and throughout post-treatment. All treatments were well-tolerated, both peroxide-containing systems exhibited appreciable color retention throughout the 18-month post-treatment period, and there were no meaningful, persistent adverse events seen with long-term follow-up. PMID- 19681256 TI - Randomized clinical trial comparing whitening strips, paint-on gel and negative control. AB - PURPOSE: To evaluate efficacy and safety of peroxide-containing whitening strips and a paint-on gel relative to a non-peroxide experimental control. METHODS: After informed consent, 52 healthy adults in Shanghai, China were randomized to one of three treatment groups: 6% hydrogen peroxide whitening strips (Crest Whitestrips), 5.9% hydrogen peroxide paint-on gel (Colgate Simply White), or water rinse which served as a negative experimental control. Strip use was twice daily over 7 days, while the paint-on gel and rinse were used twice daily over 14 days. Efficacy was measured from standard digital images of the maxillary anterior teeth, and safety was assessed from interview and intraoral examination. RESULTS: Whitening strips provided the greatest end-of-treatment reduction in yellowness (Deltab*), with adjusted means +/- standard errors of -1.72 +/- 0.18 for the strip group, -0.48 +/- 0.10 for the paint-on gel group, and 0.13 +/- 0.09 for the water rinse group. For DeltaL* (lightness), end-of-treatment adjusted means +/- standard errors were 1.88 +/- 0.21 for the strip group, 0.60 +/- 0.15 for the paint-on gel, and -0.10 +/- 0.18 for the negative control. Groups differed significantly (P< 0.007) with respect to Deltab* and DeltaL* at end-of treatment, as well as other color parameters. All treatments were well-tolerated. PMID- 19681257 TI - Clinical trial of tooth whitening with 6% hydrogen peroxide whitening strips and two whitening dentifrices. AB - PURPOSE: To compare tooth whitening with 6% hydrogen peroxide whitening strips and two whitening dentifrices in a 12-week randomized controlled trial at a Belgian dental school. METHODS: After informed consent, 46 healthy adults were randomly assigned to one of three strip + dentifrice treatment groups. Subjects received either 6% hydrogen peroxide whitening strips (Crest Whitestrips) and an anticavity toothpaste (Crest Cavity Protection), placebo strips and a sodium fluoride (NaF) whitening dentifrice (Mentadent Whitening Toothpaste) or placebo strips and a sodium monofluorophosphate (MFP) whitening dentifrice (Rembrandt Low Abrasion Whitening Toothpaste). Strip use (peroxide or placebo) was for 30 minutes, twice daily for 2 weeks, while dentifrice use was at least twice daily for 12 weeks. Efficacy was measured from standardized digital images of the maxillary facial tooth surfaces, while safety was evaluated from oral examination and interview. Treatments were compared after 2 weeks (strip use) and 12 weeks (dentifrice use) using analysis of covariance. RESULTS: All subjects completed the 12-week evaluation. Adjusting for baseline and age, the peroxide strip group had -2.45 Deltab*, 2.39 DeltaL*, and -0.96 Deltaa* at Week 2. Between-group comparisons demonstrated significant (P< 0.0001) reductions in yellowness and redness, and increased brightness favoring the peroxide strip group. The peroxide strip group demonstrated 95%+ color retention (Deltab* & DeltaL*) at Week 12, differing significantly (P< 0.0001) versus either of the continuously used whitening dentifrices. There were no significant (P> 0.18) differences between the whitening dentifrice groups at any timepoints. All treatments were well tolerated, with minor tooth sensitivity and oral irritation representing the most common findings. PMID- 19681258 TI - Risk assessment for Clostridium perfringens in ready-to-eat and partially cooked meat and poultry products. AB - An assessment of the risk of illness associated with Clostridium perfringens in ready-to-eat and partially cooked meat and poultry products was completed to estimate the effect on the annual frequency of illnesses of changing the allowed maximal 1-log growth of C. perfringens during stabilization (cooling after the manufacturing heat step). The exposure assessment modeled stabilization, storage, and consumer preparation such as reheating and hot-holding. The model predicted that assuming a 10- or 100-fold increase from the assumed 1-log (maximal allowable) growth of C. perfringens results in a 1.2- or 1.6-fold increase of C. perfringens-caused illnesses, respectively, at the median of the uncertainty distribution. Improper retail and consumer refrigeration accounted for approximately 90% of the 79,000 C. perfringens illnesses predicted by the model at 1-log growth during stabilization. Improper hot-holding accounted for 8% of predicted illnesses, although model limitations imply that this is an underestimate. Stabilization accounted for less than 1% of illnesses. Efforts to reduce illnesses from C. perfringens in ready-to-eat and partially cooked meat and poultry products should focus on retail and consumer storage and preparation methods. PMID- 19681259 TI - Surrogate selection for Escherichia coli O157:H7 based on cryotolerance and attachment to romaine lettuce. AB - Using nonpathogenic surrogates in place of pathogens when evaluating commercial food processing operations offers safety advantages, but their usefulness may be limited if they do not behave in the same manner in challenge situations. Nonpathogenic Escherichia coli strains were compared with E. coli O157:H7 based on cryotolerance, cell surface characteristics (hydrophobicity, zeta potential, and morphology), and attachment to lettuce. Populations for all strains were reduced less than 1 log CFU/ml over 7 days of storage at -18 degrees C. After 1 day of storage, the survival rate for E. coli ATCC 25922 was 44.3%, similar to that of E. coli O157:H7 (49%). No capsule was produced by any of the strains. E. coli O157:H7 expressed curli at both 20 and 37 degrees C, whereas E. coli ATCC 25922 expressed curli only when grown at 20 degrees C. Hydrophobicity of E. coli ATCC 25922 was 53.5%, similar to that of E. coli O157:H7 (56.2%). The zeta potentials of nonpathogenic E. coli and E. coli O157:H7 cells were -4.95 to 10.92 mV. The zeta potential of E. coli ATCC 25922 was not significantly different (P > 0.05) from that of E. coli O157:H7 at 37 degrees C and was the closest value to that of E. coli O157:H7 at 20 degrees C. E. coli ATCC 25922 exhibited the greatest attachment to lettuce among the surrogates and was not significantly different from E. coli O157:H7 (P > 0.05). Based on cryotolerance and cell surface characteristics, E. coli ATCC 25922 is a useful surrogate for E. coli O157:H7 for studies involving attachment to fresh produce. PMID- 19681260 TI - Efficacy of antimicrobial agents in lettuce leaf processing water for control of Escherichia coli O157:H7. AB - The objectives of this research were to study transfer and control of Escherichia coli O157:H7 during simultaneous washing of inoculated and uninoculated lettuce pieces and to determine the efficacy of antimicrobial agents (peroxyacetic acid, mixed peracid, and sodium hypochlorite) on reducing the transfer of E. coli O157:H7 through processing water with or without organic load. Lettuce leaf pieces (5 by 5 cm) were inoculated with a five-strain mixture of green fluorescent protein-labeled E. coli O157:H7 at 5.6 log CFU per piece. One inoculated lettuce piece was added to five uninoculated leaves during washing. Peroxyacetic acid and mixed peracid were tested at 10, 20, and 30 ppm, and chlorine was tested at 30 and 50 ppm. No organic load (liquefied lettuce leaves) and 10% organic load in processing water were compared. Without organic load, peroxyacetic acid at 30 ppm, mixed peracid at 10, 20, and 30 ppm, and chlorine at 30 and 50 ppm all significantly reduced E. coli O157: H7 in processing water by 1.83, 1.73, 1.50, 1.83, 1.34, and 1.83 log CFU/ml, respectively, compared with washing with water alone. These antimicrobials at all concentrations tested also significantly reduced transfer of the bacteria from an inoculated leaf to uninoculated leaves in the processing water by 0.96 to 2.57 log CFU per piece. A 10% organic load in the processing water reduced efficacy of antimicrobial agents. In this contaminated water, peroxyacetic acid at 10 and 20 ppm and chlorine at 30 ppm produced effects not significantly different from those of water alone. Therefore, it is important to understand the impact of organic load when validating the effectiveness of antimicrobial treatments. PMID- 19681261 TI - Survival of Escherichia coli after isoelectric solubilization and precipitation of fish protein. AB - Protein recovery for fish processing by-products utilizes extreme pH shifts for isoelectric solubilization and precipitation. The purpose of this study was to determine if Escherichia coli would survive exposure to the extreme pH shifts during the protein recovery process. Fresh rainbow trout were beheaded, gutted, and minced and then inoculated with approximately 10(9) CFU of E. coli ATCC 25922 per g, homogenized, and brought to the target pH of 2.0, 3.0, 11.5, or 12.5 by the addition of concentrated hydrochloric acid or sodium hydroxide to solubilize muscle proteins. The homogenate was blended and centrifuged to separate the lipid and insoluble components (bones, skin, insoluble protein, etc.) from the protein solution. The protein solution was subjected to a second pH shift (pH 5.5) resulting in protein precipitation that was recovered with centrifugation. Microbial analysis was conducted on each fraction (i.e., lipid, insoluble components, protein, and water) with selective and nonselective media. The sums of the surviving E. coli in these fractions were compared with the initial inoculum. The greatest total microbial reduction occurred when the pH was shifted to 12.5 (P < 0.05), i.e., a 4.4-log reduction of cells on nonselective media and a 6.0-log reduction of cells on selective media. The use of selective and nonselective media showed that there was significant (P < 0.05) injury sustained by cells exposed to alkaline treatment (pH 11.5 and 12.5) in all fractions except the insoluble fraction at pH 11.5. Increasing the exposure time or the pH may result in greater bacterial reductions in the recovered protein. PMID- 19681262 TI - Thermal inactivation of Escherichia coli O157:H7 in blade-tenderized beef steaks cooked on a commercial open-flame gas grill. AB - Beef subprimals were inoculated on the lean side with ca. 4.0 log CFU/g of a cocktail of rifampin-resistant (Rif(r)) Escherichia coli O157:H7 strains and then passed once through a mechanical blade tenderizer with the lean side facing upward. Inoculated subprimals that were not tenderized served as controls. Two core samples were removed from each of three tenderized subprimals and cut into six consecutive segments starting from the inoculated side. A total of six cores were also obtained from control subprimals, but only segment 1 (topmost) was sampled. Levels of E. coli O157:H7 recovered from segment 1 were 3.81 log CFU/g for the control subprimals and 3.36 log CFU/g for tenderized subprimals. The percentage of cells recovered in segment 2 was ca. 25-fold lower than levels recovered from segment 1, but E. coli O157:H7 was recovered from all six segments of the cores obtained from tenderized subprimals. In phase II, lean-side inoculated (ca. 4.0 log CFU/g), single-pass tenderized subprimals were cut into steaks of various thicknesses (1.91 cm [0.75 in.], 2.54 cm [1.0 in.], and 3.18 cm [1.25 in.]) that were subsequently cooked on a commercial open-flame gas grill to internal temperatures of 48.8 degrees C (120 degrees F), 54.4 degrees C (130 degrees F), and 60 degrees C (140 degrees F). In general, regardless of temperature or thickness, we observed about a 2.6- to 4.2-log CFU/g reduction in pathogen levels following cooking. These data validate that cooking on a commercial gas grill is effective at eliminating relatively low levels of the pathogen that may be distributed throughout a blade-tenderized steak. PMID- 19681264 TI - Effect of gamma radiation on the quality and shelf life of refrigerated rainbow trout (Oncorhynchus mykiss) fillets. AB - The effect of gamma radiation (0, 1, 3, and 5 kGy) on the shelf life of farmed rainbow trout (Oncorhynchus mykiss) fillets that were treated with sodium acetate and vacuum packaged and subsequently stored under refrigeration was studied by measuring microbiological, chemical, and organoleptic changes. Radiation affected populations of bacteria, namely, H2S-producing bacteria and Enterobacteriaceae (P < 0.05). Initial total viable counts of the control samples were ca. 4.41 log CFU/g, whereas the respective counts in samples irradiated at 1, 3, and 5 kGy were 3.08, 1.46, and < or = 1 log CFU/g at day 1 of refrigerated storage. The maximum count of Enterobacteriaceae reached 2.29 and 1.45 log CFU/g at the end of storage for 1 and 3 kGy, respectively, but at a 5-kGy dose no growth of Enterobacteriaceae was observed. Of the biochemical indicators, thiobarbituric acid values for irradiated trout were higher than for nonirradiated fish (P < 0.05). Sensory evaluation (taste) showed a reasonable and good correlation with bacterial populations with storage time. The results revealed that radiation at a high dose (5 kGy) might induce lipid and protein oxidation, although the growth of microorganisms was inhibited. Therefore, radiation at a low dose (3 kGy) could be used to control the microbial and safety biochemical indices of O. mykiss for up to 4 weeks at refrigerator temperature without adverse effects on quality and acceptability. PMID- 19681263 TI - Heat and acid tolerance responses of Listeria monocytogenes as affected by sequential exposure to hurdles during growth. AB - This study aimed to evaluate the effects of the level and sequence of hurdles, applied during growth, on the subsequent heat and acid tolerances of a 10-strain composite of Listeria monocytogenes. Individual strains were grown in glucose free tryptic soy broth with 0.6% yeast extract (TSBYE-G). Then cultures were mixed and inoculated in fresh TSBYE-G (0.5% NaCl, pH 7.42; control), TSBYE-G that was supplemented with 3% NaCl (3.5% NaCl in total), or TSBYE-G with pH adjusted to 6.01 or 5.04 with lactic acid and incubated at 30 degrees C for 24 h. Furthermore, the culture composite was exposed to the following five combinations of double sequential hurdles (12 h in each at 30 degrees C): NaCl then pH 6.01, NaCl then pH 5.04, pH 7.42 then NaCl, pH 5.04 then NaCl, and pH 6.01 then NaCl. The heat and acid tolerances of the culture were assessed at 57 degrees C (for 2 h) and at pH 3.5 (for 7 h), respectively, in TSBYE-G. No significant (P > or = 0.05) differences in thermotolerance were observed among cultures exposed to various stresses. In contrast, the acid resistance followed the order: pH 6.01 = NaCl > NaCl then pH 5.04 > pH 6.01 then NaCl = pH 5.04 > pH 5.04 then NaCl > pH 7.42 then NaCl > control. The results suggest that exposure of L. monocytogenes to NaCl and low pH during growth may not affect its heat (57 degrees C) tolerance, but it may increase its acid (pH 3.5) resistance, depending on the sequence and intensity of the applied stresses. PMID- 19681265 TI - Colletotrichum gloeosporioides growth-no-growth interface after selected microwave treatments. AB - To study microwave heating for potential postharvest treatments against anthracnose disease, Colletotrichum gloeosporioides growth-no-growth response after selected microwave treatments (2,450 MHz) was fitted by using a logistic regression model. Evaluated variables were power level, exposure time, presence or absence of water in the medium during treatment, and incubation-observation time. Depending on the setting, the applied power ranged from 77.2 to 435.6 W. For the experiments on dry medium (mold spores over filter paper), exposure times were 1, 2, 3, or 4 min, whereas spores dispersed in potato dextrose agar, a wet medium, had exposure times of 3, 6, or 9 s. Growth (response = 1) or no growth (response = 0) was observed after two different incubation-observation times (4 or 10 days). As expected, high power levels and long exposure times resulted in complete inhibition of C. gloeosporioides spore germination. In a number of cases (such as low power levels and short treatment times), only a delay in mold growth was observed. Scanning electron micrographs showed signs of mycelia dehydration and structural collapse in the spores of the studied mold. Cell damage was attributed to heating during microwave exposure. Reduced logistic models included variables and interactions that significantly (P < 0.05) affected mold growth, and were able to predict the growth-no-growth response in at least 83% of the experimental conditions. Microwave treatments (4 min at any of the studied power levels in dry medium, and 9 s at power levels of 30% or more for wet medium) proved effective in the inhibition of C. gloeosporioides in model systems. These no-growth conditions will be tested further on fresh fruits in order to develop feasible postharvest microwave treatments. PMID- 19681266 TI - Development of a high pressure processing inactivation model for hepatitis A virus. AB - High pressure processing (HPP) inactivation data were obtained for hepatitis A virus (HAV) suspended in buffered growth medium containing salt at either 15 or 30 g/liter. Pressures between 300 and 500 MPa were applied for treatment times of 60 to 600 s. In medium containing 15 g/liter salt, the HAV titer was reduced by approximately 1 and 2 log 50% tissue culture infectious dose units (TCID50) per ml after 600 s of treatment with 300 and 400 MPa, respectively. Under the same treatment conditions but in medium containing 30 g/liter salt, HAV was reduced by <0.50 log TCID50/ml. HAV was inactivated by >3 log TCID50/ml after treatment with 500 MPa for 300 and 360 s in medium containing 15 and 30 g/liter salt, respectively. Weibull and log-linear models were fitted to inactivation data. Individual Weibull curves generally provided a good fit at each pressure and salinity, but the curve shapes were qualitatively inconsistent between treatments, making interpolation between pressures difficult and unreliable. High variability was observed in the inactivation data, but the log-linear model described the entire data set and interpolated between specific treatment conditions. Therefore, this model was evaluated by using high pressure to treat HAV artificially inoculated into Pacific oyster (Crassostrea gigas) homogenate adjusted to 15 or 30 g/liter salinity. The log-linear model generally provided fail-safe predictions at pressures > or = 375 MPa and may aid shellfish processors wishing to incorporate HPP into an oyster processing regime. Additional inactivation data with greater reproducibility should be collected to enable expansion of the model and to increase the accuracy of its predictions. PMID- 19681267 TI - Foodborne and indicator bacteria in farmed molluscan shellfish before and after depuration. AB - Galicia's coast (northwestern Spain) is a major producer of bivalve molluscs. Over an 18-month period, the presence of Salmonella, Aeromonas, Plesiomonas shigelloides, Vibrio parahaemolyticus, and Clostridium botulinum was determined by PCR methods in mussels (22 batches) and infaunal bivalves (31 batches of clams and cockles) before and after depuration. All batches were harvested from Galician class B harvesting areas where bivalve molluscs must not exceed 4,600 Escherichia coli per 100 g of flesh and liquor in 90% of the samples. Virulence associated genes of Salmonella (invA), Aeromonas (aerA, hlyA, alt, ast, and laf), P. shigelloides (hugA), V. parahaemolyticus (tdh and trh), and C. botulinum (BoNT) were not detected. The pR72H chromosomal DNA fragment, which is conservative in V. parahaemolyticus strains, was detected in five (4.7%) samples. A number of 192 suspect isolates did not fit the description of clinical Aeromonas phenospecies, pathogenic Vibrio spp., or P. shigelloides. The effectiveness of commercial depuration in reducing bacterial indicators was also examined. E. coli was reduced to < or = 230/100 g of flesh and liquor in 90.9% of mussel lots but in only 70.9% of infaunal bivalve lots. For total coliform elimination, mussels were also more effective. Total counts significantly (P < 0.005) correlated with numbers of Pseudomonas, Aeromonas, and Vibrio. Our data indicate that Salmonella and pathogenic bacteria indigenous to estuarine environments do not appear to be significant hazards in Galician molluscan shellfish. A reason for concern, however, is that clearance of E. coli to acceptable levels was not always achieved especially in infaunal bivalves. PMID- 19681268 TI - Listeria monocytogenes in the Irish dairy farm environment. AB - Listeria monocytogenes is a potentially lethal foodborne pathogen commonly found in the environment. European Union hygiene legislation places responsibility for safety on primary production facilities, including farms, as part of a policy to introduce traceability throughout the food chain. This study aimed to determine the occurrence of L. monocytogenes in the Irish dairy farm environment and in particular the milking facility. Two hundred ninety-eight environmental samples were collected from 16 farms in the southern region of Ireland. A number of farms within the group supply raw milk to the unpasteurized milk cheese industry. The samples taken included cow feces, milk, silage, soil, water, etc. Samples were enriched in Listeria enrichment broth and incubated for 48 h, followed by plating on chromogenic agar Listeria Ottavani & Agosti and further incubation of the plates for 24 to 48 h. Presumptive L. monocytogenes isolates were purified and confirmed by PCR targeting the hly gene. Overall, 19% of the samples (57 of 298) were positive for L. monocytogenes. These were serotyped using conventional and PCR methods; serotypes 1/2a, 1/2b, and 4b made up 78% of the typeable isolates. A correlation was found between the level of hygiene standards on the farm and the occurrence of L. monocytogenes. There was little difference in the occurrence of L. monocytogenes between farms supplying milk to the unpasteurized milk cheese industry and those supplying milk for processing. This study demonstrates the prevalence of L. monocytogenes in the dairy farm environment and the need for good hygiene practices to prevent its entry into the food chain. PMID- 19681269 TI - Prevalence of Mycobacterium avium subsp. paratuberculosis in ileocecal lymph nodes and on hides and carcasses from cull cows and fed cattle at commercial beef processing plants in the United States. AB - Clinical associations between Crohn's disease in humans and Mycobacterium avium subsp. paratuberculosis (MAP) have been suggested but not confirmed. Cattle could be sources for MAP, but little information on MAP prevalence with beef has been reported. Samples of ileocecal lymph nodes and swabs of hides and carcasses from 343 animals at cull cattle slaughtering facilities and 243 animals at fed cattle slaughtering facilities across the United States were analyzed for the presence of MAP. Amplification of genetic sequences detected MAP DNA predominantly on hides and in lymph nodes of samples taken at both types of processing facilities. More than 34% of the cattle at cull cow slaughtering facilities had ileocecal lymph nodes that tested positive for MAP DNA. From these same cattle, hide prevalence was more than twofold greater than the prevalence in ileocecal lymph nodes, suggesting that cross-contamination could be occurring during transport and lairage. The prevalence of MAP DNA decreased during processing, and less than 11% of the carcasses tested positive after interventions in the cull cow processing facilities. Using standard double-decontamination and culture techniques, less than 1% of the postintervention carcasses tested positive for viable MAP at cull cow facilities. In samples from the facilities processing only fed cattle, MAP prevalence of 1% or less was detected for ileocecal lymph node, hide, and carcass samples, and viable MAP was not detected. Based on this study, fed cattle carcasses are unlikely sources of MAP, and carcasses at cull cow plants have only a slight risk for transmitting viable MAP, due to current interventions. PMID- 19681270 TI - Impact of potential changes to the current bovine spongiform encephalopathy surveillance programs for slaughter cattle and fallen stock in Japan. AB - Cattle slaughtered in Japan for human consumption, regardless of their age, have been tested for bovine spongiform encephalopathy (BSE) since October 2001. Beginning in April 2004, all fallen stock from 24 months of age also have been tested. We evaluated the impact of potential changes to the current BSE surveillance programs for both slaughter cattle and fallen stock using a simple stochastic model. We calculated the probability that a BSE-infected dairy cow, Wagyu beef animal, Wagyu-Holstein cross steer or heifer, or Holstein steer slaughtered for human consumption or arising as fallen stock would be tested and detected. Four surveillance strategies were explored for cattle slaughtered for human consumption, with the minimum age at testing set at 0, 21, 31, or 41 months. Three surveillance strategies were explored for fallen stock, with the minimum age at testing set at 24, 31, or 41 months. Increasing the minimum age of testing from 0 to 21 months for both dairy cattle and Wagyu beef cattle had very little impact on the probability that a BSE-infected animal slaughtered for human consumption would be detected. Although increasing the minimum age at testing from 21 to 31 or 41 months would lead to fewer slaughtered animals being tested, the impact on the probability of detecting infected animals would be insignificant. The probability of infected Wagyu-Holstein crosses and Holstein steers being detected at slaughter or as fallen stock would be very low under all surveillance strategies. PMID- 19681271 TI - Food protective effect of geraniol and its congeners against stored food mites. AB - The acaricidal activities of compounds derived from the oil of Pelargonium graveolens leaves against the storage food mite, Tyrophagus putrescentiae, were compared with the activity of a commercial acaricide, benzyl benzoate, in an impregnated fabric disk bioassay. Purification of the active constituent from P. graveolens was accomplished by silica gel chromatography and high-performance liquid chromatography. Structural analysis of the active constituent by 1H nuclear magnetic resonance (NMR), 13C-NMR, 1H-13C shift correlated spectroscopy NMR, and distortionless enhancement by polarization transfer NMR identified trans 3,7-dimethyl-2,6-octadien-1-ol (geraniol). Based on the 50% lethal dose values, the most toxic compounds against T. putrescentiae were geraniol (1.95 microg/cm3), which was followed by nerol (2.21 microg/cm3), citral (9.65 microg/cm3), benzyl benzoate (11.27 microg/cm3), and beta-citronellol (15.86 microg/cm3). Our results suggest that geraniol is more effective in controlling T. putrescentiae than benzyl benzoate is. Furthermore, geraniol, which is used as a flavoring for beverages, candies, ice creams, and baked goods and congeners (citral and nerol), may be useful for managing populations of T. putrescentiae. PMID- 19681272 TI - Evaluation of a new one-step enrichment in conjunction with a chromogenic medium for the detection of Cronobacter spp. (Enterobacter sakazakii) in powdered infant formula. AB - The aim of the present study was to evaluate a new one-step enrichment protocol, consisting of a combined preenrichment and enrichment broth (Cronobacter Enrichment Broth [CEB]) used in conjunction with selective-differential agar ChromID Sakazakii, to facilitate a shortened 2-day cultural method for detection of Cronobacter spp. (Enterobacter sakazakii) in powdered infant formula (PIF). The CEB was evaluated using samples artificially inoculated with low concentrations of 10 lyophilized strains, representative of the genus Cronobacter. The detection of strains was compared in parallel with the enrichment medium from ISO/TS 22964 and a recently proposed differential screening broth for the detection of Cronobacter. All of the Cronobacter strains were recovered using the CEB, and a significantly higher final bacterial concentration was obtained with the CEB than with the other enrichment broths (P < 0.01). There was no significant difference between the cell concentrations for cultures grown in CEB at 37 degrees C and those grown at 41.5 degrees C. Cronobacter was recovered from both 1/10 (50 g:450 ml) and 1/5.5 (100 g:450 ml) sample-to-broth ratios, with no significant difference observed between the final bacterial concentrations obtained from the two ratios. Further studies on a wider range of PIFs, including naturally contaminated samples, are warranted to determine if the use of this protocol may facilitate the rapid release (within 40 to 48 h) of PIF. PMID- 19681273 TI - Biofilm formation ability of Listeria monocytogenes isolates from raw ready-to eat seafood. AB - Listeria monocytogenes is of great concern as a foodborne pathogen. Many ready-to eat foods are widely contaminated with this organism and have caused listeriosis outbreaks and sporadic cases in many countries. In Japan, there is a high incidence of L. monocytogenes contamination, specifically in raw ready-to-eat seafood. Identical L. monocytogenes subtypes have been isolated repeatedly from samples of food manufactured at a given store or processing plant, and researchers suspected that certain L. monocytogenes isolates have formed biofilms at these sites. A microtiter plate biofilm formation assay was conducted, and all raw ready-to-eat seafood isolates tested were able to form biofilms to various degrees. Biofilm formation by L. monocytogenes isolates of lineage I was significantly greater (P = 0.000) than that by isolates of lineage II. However, isolates of clonal lineages formed different levels of biofilms, indicating that the ability to form a biofilm is affected positively or negatively by environmental factors. PMID- 19681274 TI - Effectiveness of bacteriophages in reducing Escherichia coli O157:H7 on fresh-cut cantaloupes and lettucet. AB - Consumption of produce contaminated with Escherichia coli O157:H7 has resulted in cases of foodborne illness. We determined the efficacy of a mixture of three E. coli O157:H7-specific bacteriophages (ECP-100) in reducing the number of viable E. coli O157:H7 on contaminated fresh-cut iceberg lettuce and cantaloupe. E. coli O157:H7 was spot inoculated on lettuce pieces (9 cm2) with a population of 3.76 log CFU/cm2, allowed to dry, and then sprayed with a control (phosphate-buffered saline) or ECP-100 to deliver 7.98 log PFU/cm2 to lettuce stored for 2 days at 4 degrees C. Cut pieces of cantaloupe were spot inoculated with E. coli O157:H7 (4.55 log CFU/ml) and treated with the control or ECP-100 (6.69 log PFU/ml), and then stored at 4 or 20 degrees C for up to 7 days. On days 0, 2, 5, and 7, cantaloupe samples were homogenized, and populations of E. coli O157:H7 were enumerated. Populations of E. coli O157:H7 on lettuce treated with ECP-100 on 0, 1, and 2 days (0.72, <0.22, and 0.58 log CFU/cm2 of lettuce) and stored at 4 degrees C were significantly (P < 0.05) lower than those treated with the control (2.64, 1.79, and 2.22 log CFU/cm2), respectively. Populations on cut cantaloupes treated with ECP-100 on days 2, 5, and 7 (0.77, 1.28, and 0.96 log CFU/ml) and stored at 4 degrees C were significantly lower than those cut cantaloupes treated with the control (3.34, 3.23, and 4.09 log CFU/ml), respectively. This study is the first to show the effectiveness of bacteriophages to reduce E. coli O157:H7 on fresh-cut lettuce and cantaloupes. PMID- 19681275 TI - Efficacy of detergents and fresh produce disinfectants against microorganisms associated with mixed raw vegetables. AB - Efficacy of commercial detergent and disinfectants to eliminate microorganisms associated with fresh vegetables eaten raw in Iran, including radish, parsley, basil, coriander (cilantro), Allium porrum (leek), and peppermint were studied. The raw vegetables were subjected to a triple wash treatment of washing in tap water for mud removal, washing in water containing a detergent (dishwashing liquid) or disinfectant individually, and rinsing in tap water. The population of total mesophilic microbes on the surface of untreated vegetables ranged from 10(5) to 10(6) CFU/g. Washing in tap water or treatment with detergent (333 ppm for 10 min) or benzalkonium chloride (92 ppm for 15 min) reduced the total microbial count, most probable number (MPN) of coliforms, MPN of fecal coliforms, and MPN of fecal streptococci by about 1.2 to 2.3 log. No significant differences in microbial populations were found on vegetables after decontamination with tap water, detergent, or benzalkonium chloride (P > 0.05). Treatments with peracetic acid (100 ppm for 15 min) and hydrogen peroxide (133 ppm for 30 min) reduced the total mesophilic microbial counts by about 2.8 log. The microbial reductions with calcium hypochlorite (300 ppm for 15 min) and combined hydrogen peroxide and silver ion (133 ppm for 30 min) were significantly higher than those obtained after rinsing in tap water or after detergent or benzalkonium chloride wash (P < 0.05). Pretreatment with detergent slightly enhanced the efficacy of all decontamination treatments, but results were not significantly different from those obtained after individual application of disinfectants. PMID- 19681277 TI - Development of primers for detection of heat-treated cetacean materials in porcine meat and bone meal. AB - The feed ban introduced after the detection of the first case of bovine spongiform encephalopathy in 2001 in Japan has been modified to allow some of the previously prohibited animal materials to be used in animal feed. Recently, porcine materials were allowed to be used in feed for pigs, poultry, and fish. Materials from other mammals, including whales, remain prohibited. In the absence of a method to detect the prohibited whale materials in porcine materials, there is a possibility that the whale materials are being used for feed for pigs, poultry, and fish. To detect illegal use of whale materials mixed with porcine materials, we have developed PCR primers specific to a group of most cetacean species, using a computer-based method we developed previously. The primer sets were capable of detecting whale meat meal that had been autoclaved at 133 degrees C for up to 20 min. The detection limit of whale material in porcine meat and bone meal was 0.1%. PMID- 19681276 TI - Specific PCR detection of Arcobacter butzleri, Arcobacter cryaerophilus, Arcobacter skirrowii, and Arcobacter cibarius in chicken meat. AB - An enrichment PCR assay using species-specific primers was developed for the detection of Arcobacter butzleri, Arcobacter cryaerophilus, Arcobacter skirrowii, and Arcobacter cibarius in chicken meat. Primers for A. cryaerophilus, A. skirrowii, and A. cibarius were designed based on the gyrA gene to amplify nucleic acid fragments of 212, 257, and 145 bp, respectively. The A. butzleri specific primers were designed flanking a 203-bp DNA fragment in the 16S rRNA gene. The specificity of the four primer pairs was assessed by PCR analysis of DNA from a panel of Arcobacter species, related Campylobacter, Helicobacter species, and other food bacteria. The applicability of the method was then validated by testing 42 fresh retail-purchased chicken samples in the PCR assay. An 18-h selective preenrichment step followed by PCR amplification with the four Arcobacter primer sets revealed the presence of Arcobacter spp. in 85.7% of the retail chicken samples analyzed. A. butzleri was the only species present in 50% of the samples, and 35.7% of the samples were positive for both A. butzleri and A. cryaerophilus. A. skirrowii and A. cibarius were not detected in any of the chicken samples analyzed. The enrichment PCR assay developed is a specific and rapid alternative for the survey of Arcobacter contamination in meat. PMID- 19681278 TI - High hydrostatic pressure for development of vaccines. AB - Disease management in the food industry is complex and includes use of good hygienic practices, antimicrobials, and immunization. Vaccines are available against many, but not all, disease agents affecting animals reared for human food. Fewer vaccines are currently licensed and widely available for human foodborne pathogens. Increased resistance to antimicrobials provides additional impetus to develop new vaccines. In addition to the need for new vaccines, new methods of vaccine production are desired. Some current methods of vaccine production can involve use of hazardous chemicals, provide inconsistent results, or present risk to vaccine recipients with certain allergies. The efficacy of high hydrostatic pressure (HHP) for inactivation of a variety of foodborne pathogenic microorganisms has been well established, and some of these microorganisms have been demonstrated to retain immunogenic properties, suggesting HHP may have application for the development of vaccines. Studies on the effect of HHP on infectivity and immunogenicity of various viruses, a protozoan parasite, and one bacterial species are presented. Control of several of these pathogens is important for animal health and economic stability in several sectors of the food industry. The research to date on the potential for vaccine development by HHP is presented. PMID- 19681279 TI - A novel approach to enhance food safety: industry-academia-government partnership for applied research. AB - An independent collaborative approach was developed for stimulating research on high-priority food safety issues. The Fresh Express Produce Safety Research Initiative was launched in 2007 with $2 million in unrestricted funds from industry and independent direction and oversight from a scientific advisory panel consisting of nationally recognized food safety experts from academia and government agencies. The program had two main objectives: (i) to fund rigorous, innovative, and multidisciplinary research addressing the safety of lettuce, spinach, and other leafy greens and (ii) to share research findings as widely and quickly as possible to support the development of advanced safeguards within the fresh-cut produce industry. Sixty-five proposals were submitted in response to a publicly announced request for proposals and were competitively evaluated. Nine research projects were funded to examine underlying factors involved in Escherichia coli O157:H7 contamination of lettuce, spinach, and other leafy greens and potential strategies for preventing the spread of foodborne pathogens. Results of the studies, published in the Journal of Food Protection, help to identify promising directions for future research into potential sources and entry points of contamination and specific factors associated with harvesting, processing, transporting, and storing produce that allow contaminants to persist and proliferate. The program provides a model for leveraging the strengths of industry, academia, and government to address high-priority issues quickly and directly through applied research. This model can be productively extended to other pathogens and other leafy and nonleafy produce. PMID- 19681280 TI - A novel approach to investigate the uptake and internalization of Escherichia coli O157:H7 in spinach cultivated in soil and hydroponic medium. AB - Internalization of Escherichia coli O157:H7 into spinach plants through root uptake is a potential route of contamination. A Tn7-based plasmid vector was used to insert a green fluorescent protein gene into the attTn7 site in the E. coli chromosome. Three green fluorescent protein-labeled E. coli inocula were used: produce outbreak O157:H7 strains RM4407 and RM5279 (inoculum 1), ground beef outbreak O157:H7 strain 86-24h11 (inoculum 2), and commensal strain HS (inoculum 3). These strains were cultivated in fecal slurries and applied at ca. 10(3) or 10(7) CFU/g to pasteurized soils in which baby spinach seedlings were planted. No E. coli was recovered by spiral plating from surface-sanitized internal tissues of spinach plants on days 0, 7, 14, 21, and 28. Inoculum 1 survived at significantly higher populations (P < 0.05) in the soil than did inoculum 3 after 14, 21, and 28 days, indicating that produce outbreak strains of E. coli O157:H7 may be less physiologically stressed in soils than are nonpathogenic E. coli isolates. Inoculum 2 applied at ca. 10(7) CFU/ml to hydroponic medium was consistently recovered by spiral plating from the shoot tissues of spinach plants after 14 days (3.73 log CFU per shoot) and 21 days (4.35 log CFU per shoot). Fluorescent E. coli cells were microscopically observed in root tissues in 23 (21%) of 108 spinach plants grown in inoculated soils. No internalized E. coli was microscopically observed in shoot tissue of plants grown in inoculated soil. These studies do not provide evidence for efficient uptake of E. coli O157:H7 from soil to internal plant tissue. PMID- 19681281 TI - Effect of route of introduction and host cultivar on the colonization, internalization, and movement of the human pathogen Escherichia coli O157:H7 in spinach. AB - Human pathogens can contaminate leafy produce in the field by various routes. We hypothesized that interactions between Escherichia coli O157:H7 and spinach are influenced by the route of introduction and the leaf microenvironment. E. coli O157:H7 labeled with green fluorescent protein was dropped onto spinach leaf surfaces, simulating bacteria-laden raindrops or sprinkler irrigation, and survived on the phylloplane for at least 14 days, with increasing titers and areas of colonization over time. The same strains placed into the rhizosphere by soil infiltration remained detectable on very few plants and in low numbers (10(2) to 10(6) CFU/g fresh tissue) that decreased over time. Stem puncture inoculations, simulating natural wounding, rarely resulted in colonization or multiplication. Bacteria forced into the leaf interior survived for at least 14 days in intercellular spaces but did not translocate or multiply. Three spinach cultivars with different leaf surface morphologies were compared for colonization by E. coli O157:H7 introduced by leaf drop or soil drench. After 2 weeks, cv. Bordeaux hosted very few bacteria. More bacteria were seen on cv. Space and were dispersed over an area of up to 0.3 mm2. The highest bacterial numbers were observed on cv. Tyee but were dispersed only up to 0.15 mm2, suggesting that cv. Tyee may provide protected niches or more nutrients or may promote stronger bacterial adherence. These findings suggest that the spinach phylloplane is a supportive niche for E. coli O157:H7, but no conclusive evidence was found for natural entry into the plant interior. The results are relevant for interventions aimed at minimizing produce contamination by human pathogens. PMID- 19681282 TI - Interaction of Escherichia coli O157:H7 with leafy green produce. AB - Enterohemorrhagic Escherichia coli (EHEC) is a foodborne pathogen responsible for human diarrheal disease. EHEC lives in the intestinal tract of cattle and other farm and wild animals, which may be the source of environmental contamination particularly of agricultural fields. Human infections are associated with consumption of tainted animal products and fresh produce. How the bacteria interact with the plant phyllosphere and withstand industrial decontamination remain to be elucidated. The goals of the present study were to investigate the environmental conditions and surface structures that influence the interaction of EHEC O157:H7 with baby spinach and lettuce leaves in vitro. Independently of the production of Shiga toxin, EHEC O157:H7 colonizes the leaf surface via flagella and the type 3 secretion system (T3SS). Ultrastructural analysis of EHEC-infected leafy greens revealed the presence of flagellated bacteria, and mutation of the fliC flagellin gene in EHEC EDL933 rendered the bacteria significantly less adherent, suggesting the involvement of flagella in the bacteria-leaf interaction. EDL933 mutated in the escN (ATPase) gene associated with the function of the T3SS but not in the eae (intimin adhesin) gene required for adherence to host intestinal cells had significantly reduced adherence compared with that of the parental strain. The data suggest a compelling role of flagella and the T3SS in colonization of leafy green produce. Colonization of salad leaves by EHEC strains may be a strategy that ensures survival of these bacteria in the environment and allows transmission to the human host. PMID- 19681283 TI - Inactivation of Escherichia coli O157:H7 and natural microbiota on spinach leaves using gaseous ozone during vacuum cooling and simulated transportation. AB - The aim of this study was to integrate an ozone-based sanitization step into existing processing practices for fresh produce and to evaluate the efficacy of this step against Escherichia coli O157:H7. Baby spinach inoculated with E. coli O157:H7 (approximately 10(7) CFU/g) was treated in a pilot-scale system with combinations of vacuum cooling and sanitizing levels of ozone gas (SanVac). The contribution of process variables (ozone concentration, pressure, and treatment time) to lethality was investigated using response-surface methodology. SanVac processes decreased E. coli O157:H7 populations by up to 2.4 log CFU/g. An optimized SanVac process that inactivated 1.8 log CFU/g with no apparent damage to the quality of the spinach had the following parameters: O3 at 1.5 g/kg gas mix (935 ppm, vol/vol), 10 psig of holding pressure, and 30 min of holding time. In a separate set of experiments, refrigerated spinach was treated with low ozone levels (8 to 16 mg/kg; 5 to 10 ppm, vol/vol) for up to 3 days in a system that simulated sanitization during transportation (SanTrans). The treatment decreased E. coli populations by up to 1.4 log CFU/g, and the optimum process resulted in a 1.0-log inactivation with minimal effect on product quality. In a third group of experiments, freshly harvested unprocessed spinach was inoculated with E. coli O157:H7 and sequentially subjected to optimized SanVac and SanTrans processes. This double treatment inactivated 4.1 to > or = 5.0 log CFU/g, depending on the treatment time. These novel sanitization approaches were effective in considerably reducing the E. coli O157: H7 populations on spinach and should be relatively easy to integrate into existing fresh produce processes and practices. PMID- 19681284 TI - Association of Escherichia coli O157:H7 with filth flies (Muscidae and Calliphoridae) captured in leafy greens fields and experimental transmission of E. coli O157:H7 to spinach leaves by house flies (Diptera: Muscidae). AB - The recent outbreak of Escherichia coli O157:H7 infection associated with contaminated spinach led to an investigation of the role of insects, which frequent fields of leafy greens and neighboring rangeland habitats, in produce contamination. Four leafy greens fields adjacent to cattle-occupied rangeland habitats were sampled using sweep nets and sticky traps. Agromyzid flies, anthomyiid flies, and leafhoppers were caught consistently in both rangeland and leafy greens production fields at all sites. An unexpected number of flies (n = 34) in the Muscidae and Calliphoridae families (known as filth flies because of their development in animal feces) were caught in one leafy greens field. A subset of these filth flies were positive (11 of 18 flies) for E. coli O157:H7 by PCR amplification using primers for the E. coli O157:H7-specific eae gene. Under laboratory conditions, house flies were confined on manure or agar medium containing E. coli O157:H7 tagged with green fluorescent protein (GFP) and then tested for their capacity to transfer the microbes to spinach plants. GFP-tagged bacteria were detected on surfaces of 50 to 100% of leaves examined by fluorescence microscopy and in 100% of samples tested by PCR. These results indicate that flies are capable of contaminating leafy greens under experimental conditions and confirm the importance of further investigation of the role of insects in contamination of fresh produce. PMID- 19681285 TI - Impact of preinoculation culture conditions on the behavior of Escherichia coli O157:H7 inoculated onto romaine lettuce (Lactuca sativa) plants and cut leaf surfaces. AB - Inoculum preparation methods can impact growth or survival of organisms inoculated into foods, thus complicating direct comparison of results among studies. The objective of this study was to evaluate preinoculation culture preparation for impact on Escherichia coli O157:H7 inoculated onto leaves of romaine lettuce plants and cut leaf surfaces. E. coli O157:H7 was grown quiescently or shaken at 15, 25, or 37 degrees C to different growth phases in tryptic soy or M9 minimal salts broth or agar. Cells were harvested, washed, and suspended in 0.1% peptone, Milli Q water, or well water and refrigerated for 0 or 18 h. Prepared inoculum was spotted onto cut romaine lettuce (10 microl; 3 x 10(4) CFU/10 g) or onto romaine lettuce plants (20 microl; 3 x 10(6) CFU per leaf). Cut lettuce was sealed in 100-cm2 bags (made from a commercial polymer film) and incubated at 5 or 20 degrees C. Lettuce plants were held at 23 degrees C for 24 h. For all tested conditions, levels of E. coli O157:H7 increased at 20 degrees C on cut lettuce and decreased on cut lettuce stored at 5 degrees C or on leaves of lettuce plants. At 20 degrees C, preinoculation culture conditions had little impact on growth of E. coli O157:H7 on cut lettuce. However, survival at 5 degrees C was significantly better (P < 0.05) for cultures grown at 15 or 37 degrees C in minimal medium and to late stationary phase. Impact of preinoculation handling on survival on lettuce plants was less clear due to relatively high standard deviations observed among samples. PMID- 19681287 TI - Microbial antagonists of Escherichia coli O157:H7 on fresh-cut lettuce and spinach. AB - Fresh-cut lettuce and spinach can become contaminated with pathogens at numerous points from the field to the retail market. Natural microflora present on fresh produce may help reduce the pathogen load. The objective of this study was to isolate natural microflora from fresh-cut iceberg lettuce and baby spinach and to determine whether these bacteria were antagonistic toward Escherichia coli O157:H7. Samples were collected under conditions that mimicked actual practices between production and retail sale. Evidence of naturally occurring microorganisms on fresh lettuce (295 isolates) and spinach (200 isolates) and of possible antagonistic activity toward E. coli O157:H7 was documented. Inhibitory activity by several isolates was due to either acid production or antimicrobial peptides. Bacteria with inhibitory activity were isolated from every step in the processing and handling of the fresh-cut iceberg lettuce and baby spinach. PMID- 19681286 TI - Use of the systems approach to determine the fate of Escherichia coli O157:H7 on fresh lettuce and spinach. AB - Lettuce and spinach inoculated with Escherichia coli O157:H7 were processed and handled in ways that might occur in commercial situations, including variations in holding times before and after product cooling, transportation conditions and temperatures, wash treatments, and product storage temperatures and times. Populations of background microflora and E. coli O157:H7 were enumerated after each step in the system. Data analysis was done to predict response variables with a combination of independent categorical variables. Field temperature, time before cooling, and wash treatment significantly affected E. coli O157:H7 populations on both products. The lowest populations of E. coli O157:H7 were encountered when precool time was minimal, lettuce was washed with chlorine, and storage temperature was 4 degrees C. For lettuce, field and transportation temperature were not important once the storage period started, whereas after 2 days E. coli O157:H7 populations on packaged baby spinach were not affected by field temperature. On chopped iceberg lettuce and whole leaf spinach that was packaged and stored at 4 degrees C, E. coli O157:H7 contamination could still be detected after typical handling practices, although populations decreased from initial levels in many cases by at least 1.5 log units. In abusive cases, where populations increased, the product quality quickly deteriorated. Although E. coli O157:H7 levels decreased on products handled and stored under recommended conditions, survivors persisted. This study highlights practices that may or may not affect the populations of E. coli O157:H7 on the final product. PMID- 19681288 TI - Factors impacting the regrowth of Escherichia coli O157:H7 in dairy manure compost. AB - The environmental variables affecting Escherichia coli O157:H7 regrowth in dairy manure compost were investigated. Factors evaluated were moisture content, strain variation, growth medium of inoculum, level of background microflora and inoculum, different days of composting, and acclimation at room temperature. A mathematical model was applied to describe E. coli O157 regrowth potential in compost. Repopulation occurred in autoclaved compost with a moisture content as low as 20% (water activity of 0.986) in the presence of background microflora of 2.3 to 3.9 log CFU/g. The population of all three E. coli O157 strains increased from ca. 1 to 4.85 log CFU/g in autoclaved compost, with the highest increase in the spinach-outbreak strain. However, E. coli O157 regrowth was suppressed by background microflora at ca. 6.5 log CFU/g. By eliminating acclimation at room temperature and increasing the inoculum level to ca. 3 log CFU/g, E. coli O157:H7 could regrow in the presence of high levels of background microflora. E. coli O157:H7 regrowth in the autoclaved compost collected from the field study was evident at all sampling days, with the population increase ranging from 3.49 to 6.54 log CFU/g. The fate of E. coli O157:H7 in compost was well described by a Whiting and Cygnarowicz-Provost model, with R2 greater than 0.9. The level of background microflora was a significant factor for both growth and death parameters. Our results reveal that a small number of E. coli O157 cells can regrow in compost, and both background microflora and moisture content were major factors affecting E. coli O157:H7 growth. PMID- 19681289 TI - Post-traumatic lesions of the aortic isthmus. AB - Lesions of the isthmus are the most frequent among post-traumatic lesions of the thoracic aorta (LTA): almost always secondary to closed thoracic traumas (road accidents, falls, crushing, and explosions), they are rarely iatrogenic (operatory catheterisms) or caused by penetrating wounds. In the review of the literature concerned in the report, from the analysis of 89 bibliographic sources, we note that the etiopathogenesis and the pathophysiology of the LTA still entail a very high immediate mortality, but we also note that, in recent years, remarkable improvements have been made not only in prevention, first-aid, diagnostic definition and in the understanding of the development of the LTA, but above all in therapeutic results. The correct use of the conservative approach, particularly in the immediately post-traumatic phases, the increasingly wide spread use of endovascular exclusion (T-EVAR), even if not without numerous technical difficulties, and the further improvement of open surgery, currently make it possible to guarantee the individual patient the treatment that can offer the best probabilities of success, at least immediately. Final development, and a more complete and rigorous assessment of the medium and long term results of TEVAR will allow the formulation of therapeutic strategies that are even better defined and increasingly simple to implement, on the basis of algorithms, such as the one proposed by the Authors. PMID- 19681290 TI - Significance of super-extended (D3) lymphadenectomy in gastric cancer surgery. AB - BACKGROUND: The extension of lymphadenectomy is a matter of debate in gastric cancer surgery. The purpose of the present study was to analyse our experience on D3 lymphadenectomy in the treatment of gastric cancer with special reference to post-operative morbidity and mortality, incidence of para-aortic nodal metastases and long-term prognosis. METHODS: The results of 201 patients who underwent potentially curative gastrectomy with D3 lymphadenectomy for non-metastatic gastric adenocarcinoma at the First Department of General Surgery, University of Verona, from January 1988 to December 2004, were analysed statistically. The analysis did not include gastric stump and linitis plastica type tumors. RESULTS: Twenty-six out of the 201 patients (12.9%) showed para-aortic nodal metastases. Para-aortic node involvement was significantly higher in upper third tumors (29.1%) with respect to middle (6.1%) and lower third (7.5%) (P<0.001). Sixty-two patients (30.8%) developed post-operative complications with pulmonary affections (7%), pancreatic fistulas (4.5%) and abdominal abscesses (4.5%) as the most frequently observed complaints. In-hospital mortality was 1.5%. Overall 5-year survival rate for R0-patients was 53.6%. Considering survival in relation to nodal involvement, interestingly, patients with non-regional lymph node metastases (M1a) showed a slightly better prognosis with respect to pN3 patients (3-year survival: 13.9% and 19.4% for pN3 and M1a classes, respectively). CONCLUSIONS: D3 lymphadenectomy should be considered in curative surgery for advanced gastric cancer, especially for upper third tumors, with an acceptable morbidity and no increase in mortality. Further studies with a larger number of patients are required to confirm its prognostic value. PMID- 19681291 TI - Main bile duct carcinoma management. Our experience on 38 cases. AB - BACKGROUND: Cholangiocarcinoma (CC) is rare malignant tumors arising from cells of the biliary tract. It presents some difficulties to diagnose, and is associated with a high mortality. Traditionally extrahepatic CC is divided into klatskin tumors, intermediate tract and distal or iuxtapapillar tumors according to its location within the biliary tree. CT RM, PET may provide useful diagnostic information in those patients. Surgical resection is the only chance for cure, with results depending on selected patients and careful surgical technique. Liver transplantation could offer long-term survival in selected patients when combined with chemotherapy. Chemotherapy, radiation therapy, and external drainage remain as the only treatment for inoperable patients. MATERIAL AND METHODS: The authors report their experience since 1997 inherent to 38 cases of extrahepatic CC, 21 of which were treated by surgery: their outcome has been evaluated. RESULTS: Surprisingly four of them (2 with intermediate tract tumor and 2 with distal tract tumors) are still alive and apparently disease-free after 5 years since surgery. Moreover another one patient with papillar tumor has reached 5 years survival despite has undergone surgery two times. CONCLUSION: Surgery remains the best chance for long-term survival, and lymph node status is the most important prognostic factor following R0 resection. PMID- 19681293 TI - [Chron's disease and cancer]. AB - INTRODUCTION: The patients with Crohn's disease (CD) have a greater risk of gastrointestinal and hemopoietic cancers compared with background population. Identify groups of people affected by CD with exposition to factors that play a role in the development of cancer could be useful to plan correct diagnostic and therapeutic methods. METHODS: A reaserch on "pubmed" with "Risk factors for cancer in patients with Crohn's disease", "Cancer and Crohn's disease" Crohn's disease and cancer risk" as key-words was conducted. We identified 35 studies and analized also references reported inside each single work. RESULTS: Patients with CD have a higher risk of colorectal cancer (CRC). The risk of colon cancer alone was found to be increased, with no significant increase in the risk of rectal cancer. Factors that play a role in the development of CRC are: diagnosis of CD before 25-years-old, duration of disease, severity of inflammation, habit to cigarette's smoke, a familial history of CRC and immunosuppressive therapies. The risk of small bowel cancer was found to be increased, even if overall risk remains low. Although the role of immunosuppressive therapies remain to clarify yet, patients with CD have a higher risk of development lymphoma compared with background population. CONCLUSION: Patients with CD are at high risk of large bowel, small bowel, extraintestinal and hemopoietic cancers. Selected patients with extensive colonic disease, which has been present from a young age, are at high risk of cancer and should be candidates for colonscopic surveillance. PMID- 19681292 TI - [Role of intragastric air filled ballon (Heliosphere Bag) in severe obesity. Personal experience]. AB - INTRODUCTION: Obesity leads to serious health consequences, therefore many strategies are recommended for preventing or curing this emerging problem. Treatments are various: diet, physical activity, psychotherapy, drugs and bariatric surgery. In order to try to improve the tolerance of intragastric balloons, a new device inflated with air to improve weight loss was developed in 2004 (Heliosphere BAG). We report our personal experience about this tool. MATERIAL AND METHODS: Between January 2005 and December 2007, in our unit, 50 intragastric air filled insertion were performed under analgosedation and endoscopic control. The balloon was removed (24 hours) in two patients (4%) for acute intolerance. In other 2 patients (4%) the balloon was easily removed after 5 months because of premature desuflation, radiologically confirmed. The remnant 46 balloons were removed after six months. We evaluated efficacy, tolerance and the safety of this procedure. RESULTS: Forty one women and 9 men, with a mean age of 38.1 years (range 18-62), mean basal BMI of 39.8 (range 28-64) were included, after providing informed consent. Weight and BMI loss were evaluated on all patients. BMI decreased 5.9%, weight loss was 16.8 kg. Tolerance was very good, limited only to some dispeptic symptoms during the first 2 days after insertion. No serious technical problems were noted at balloon insertion. Balloon removal was very simple after correct desuflation after the conclusion of learning curve (10 procedures). DISCUSSION: The aim to treat obesity before bariatric surgery is based on reduction of bariatric surgical risks, general surgical risks and the prevalence of cardiovascular diseases, diabetes, musculoskeletal disorders and some cancers. CONCLUSIONS: The intragastric air filled balloon showed an acceptable profile of efficacy, good tolerance and improvement of comorbidities after 6 months. PMID- 19681294 TI - [Preliminary experience on the application of metallic stents for treatment of colorectal malignant stenosis]. AB - OBJECTIVE: Application of SEMS in treating colorectal obstruction caused by both intrinsic and extrinsic tumours. METHODS: From December 2007 to February 2008 two patient underwent colorectal stenting. The first patient was affected by sigma neoplasia with multiple lung and liver metastases; the second one had a distal colonic obstruction caused by pelvic relapse of endometrial adenocarcinoma. RESULTS: In both patients successful decompression, defined as complete relief of bowel obstruction as judged by clinical symptoms and radiographic observation, was achieved. The first patient died 1 month later for disease progression after the I cycle with Capecitabine. The second patient is undergoing the II cycle with Adriamicina and Cisplatino. DISCUSSION: In our experience no precocious or posthumous complications were observed and we evaluated that SEMSs are useful in both intrinsic and extrinsic colorectal malignancies. CONCLUSIONS: SEMSs allow a rapid decompression, reduce the number of emergency surgical procedures--and also the need for stomas--in poor general condition patients, achieving a better quality of life for patient with a short estimated life and a one-stage elective surgery for patient with resectable disease. PMID- 19681295 TI - [The colonoscoy in elderly patients]. AB - AIM: The increase in the average life led an increase of diseases, acute and chronic, not only cardiovascular and respiratory system, but also gastrointestinal. Likewise we see an increase in neoplastic lesions, sometimes diagnosed in advanced stage of disease, not susceptible to radical therapy. The purpose of this work is to evaluate the usefulness, reliability and security of colonoscopy in elderly patients, age 75 years or more. MATHERIALS END METHODS: We analysed 2407 colonoscopies, performed from January 2005 to December 2006 in Unit of Digestive Endoscopy of Gastrointestinal Surgery ("Federico II" University of Naples) and in Unit of Digestive Endoscopy of "Villa Maria Clinica" (Mirabella Eclano). Of these four hundred and sixty-nine patients (19.5%), 276 men and 139 women, were 75-years-old or more. Were considered indications to endoscopy completeness of the examination, findings, presence of any complications. In all patients were performed premedication with midazolam, 2,5-5 mg and floroglucina biidrata, 40-80 mg. The patients were constantly monitored with pulsiossimetro. RESULTS: The colonoscopy was completed in 432/469 patients (92%). In 19 cases (4%) there was a neoplastic stenosis; other reasons for the failure of cecal intubation were the poor intestinal preparation (2.5%) and intolerance of the patient (1.5%). Were excluded patients in follow-up to previous surgery or polypectomy (33.5%). Other indications in the investigation were, in descending order, abdominal pain (32.8%), presence of anaemia (22.4%), rectal bleeding (19.2%), diarrhea or constipation (18.4%), presence of abdominal mass (72%). There were no complications observed, or local (perforation/bleeding), or general (cardio-respiratory or neurological). Most frequent findings were: diverticoular disease (34.4%), polyps (22.4%), cancer (12.8%), find colitis also aspecific (10.4%), hemorrhoids (8.8%). In 11.2% of patients colonoscopy was negative. CONCLUSION: The endoscopic examination of the gastrointestinal tract is now practiced daily in almost all hospitals. The advanced age of patient, with the possible presence of diseases associated is not, in our opinion, a controindication. Although present in the literature trials that emphasize the possibility of a higher incidence of complications in elderly patients, our experience shows how this control is quite safe, if executed in appropriate and correct manner, thanks also to appropriate preparation/sedation and monitoring of the patient. Compared to other diagnostic methods, such as air contrast barium enema, the computed tomography and virtual colonoscopy, "traditional" colonoscopy has the undoubted and decisive advantage of being able to run biopsies, and complete removal of the lesion, as in the case of polypectomy. In our case we have seen a high number of elderly patients, well 469/2407, equivalent to 19.5%. A neoplastic lesion was found in 165 patients (35.2%) and of these over 80% has successfully received curative treatment (surgery or polipectomy). PMID- 19681297 TI - Hemoperitoneum caused by the rupture of a giant ovarian teratoma in a 9-year-old female. Case report and literature review. AB - The Authors report the case of a 9-year-old girl suffering from acute abdominal pain, combined with mild anaemia (Hb 10.9 g/dL), leukocytosis (24.3 x 10(3) cells/dL), and a large palpable mass in the upper left quadrant. The child underwent an appendectomy 20 days before the admission to our Department. The operation performed in urgency, as well as the removal of a bulky mass situated in the left flank and the right ovary whence it arose, made it clear that abdominal signs and symptoms were caused by the twisting and rupturing of a neoformation, that would hence cause an impressive hemoperitoneum. The histopathologic examination showed a three-germ layer mature mixed teratoma. Clinical, radiologic and biochemical test (alpha-FP, beta-hcG) performed in a postoperative 2 months follow-up revealed no residual disease. PMID- 19681296 TI - Early intestinal morphological changes following benzalkonium chloride treatment in a rat model of short bowel syndrome. AB - BACKGROUND: The aim of this study was to assess early morphological changes of the residual small intestine 30 days after application of Benzalkonium Chloride (BAC) in a rat model of short bowel syndrome (SBS). MATERIALS AND METHODS: Twenty nine Wistar rats (260 +/- 20 g) underwent 80% midsmall bowel resection with end to end anastomosis. In group 1 (n=14) BAC solution 0.1% was applied to a 2 cm segment of jejunum, and in group 2 (n=15) normal saline was applied in a similar manner and the rats were sacrificed 30 days after operation. Specimens for histological examination were obtained initially and at sacrifice. RESULTS: In the BAC treated jejunal segment (group 1), a statistically significant increase (p< 0.05) was noted in villous height by 33.2%, in crypt depth by 26.4%, in muscle thickness by 26%, 109.6% in intestinal diameter, and 20% in total intestinal length, compared to group 2. CONCLUSIONS: BAC application to the serosal surface of rat's jejunum in SBS is a simple method that within only 4 weeks can topically augment the natural adaptation process noticed following intestinal resection. Further research with a tapering technique performed in sequence is suggested, to prevent possible problems associated with pseudoobstruction in the long term. PMID- 19681298 TI - [A singular bullet run in a firearm wound in the neck. A case report]. AB - The paper deals with the peculiarities of a firearm wound regarding the dynamic of the accident and particularly considering the path followed by the bullet. The patient's firearm wound is described on the basis of the instrumental examinations done. These last give us a precious help to confirm the thesis, already guessed on the clinical examination of an introduction and path followed through the digestive tract, and to show the dynamics and the still positions of the bullet inside the patient's body. The bullet was expelled spontaneously by defecation. After control of no internal damages this firearm wound has been treated simply as a normal tip wound. PMID- 19681299 TI - [Diagnosis of synovial sarcoma of the knee accidentally revealed by trauma. Role of ultrasound. Differential diagnosis by scar-hematoma]. AB - Synovial sarcoma accounts for 8-10% of all of the soft tissue sarcomas; it's characterized by high risk of local relapse, even after surgical complete excision, deceiving onset ed slowly growth. Generally arising in the contest of joint or from immediately surrounding anatomical sites, first of all affecting inferior limbs (2/3), a sarcoma of the knee, elective anatomical site, is described, accidentally diagnosed after traumatic event. Exhibiting a very poor 5 year survey, (55%), related to dimension, distal or proximal arising, necrosis rate and grading, it's the most fequently soft tissue malignancy misdiagnosed with benign neoplasms, such as Baker cyst or villonodular pigmented synovitis, considering its deceiving macroscopic and chronological features; the differential diagnosis seems to be very hard, relying on histhological biopsy. Though the normal conventional x-ray finding, clinical examination and anamnesis have suggested in our patient the ultrasonographic investigation in urgency, which revealed the suspected nature of the lesion, which must be distinguished by hematoma, much more frequent thraumatic pathology, inducing severe prognosis in false negative cases. PMID- 19681300 TI - Observation and participation. PMID- 19681301 TI - A human factors analysis of cardiopulmonary bypass machines. AB - The practice of cardiovascular surgery demands daily interface with sophisticated technologies including most commonly the cardiopulmonary bypass (CPB) machine. Although other industries have recognized the importance of considering human factors in the design of machines to reduce operator error, the evolution of the CPB machine over the past half-century has been characterized by incremental modifications of component parts with attention principally to mechanical efficiency and biocompatibility, but with little awareness of the impact of design changes on the human user. As a first step in the redesign of a safer pump, systematic observations of perfusionists during 10 adult and pediatric cases were conducted by staff cardiac surgeons and by human factors experts. Observations were classified according to accepted ergonomics principles. Perfusionists also performed usability evaluations and provided feedback concerning the design and functionality of bypass machines. Problems identified clustered around several usability themes. Issues with displays (8% of total comments) included location, legibility, format, and integration. Multiple problems with controls (11% of total comments) including location, sensitivity, and shape were identified, as were issues with audible alarms (6% of total comments). Component integration (14% of comments) and work-space design (21% of comments) were suboptimal as well. Procedural and communication issues (21% of comments) related to pump utilization, rather than pump design, were also identified, which stemmed from a lack of standardized operating room practices. Clinical issues (14% of comments) surrounding care of the patient were also identified but were not further analyzed, because these comments did not pertain to the design of the perfusion pump. Our observations confirmed the hypothesis that opportunities exist to incorporate usability and ergonomics insights into CPB machine design to optimize the human/technology interface. Such fundamental design considerations may improve the safety of the conduct of CPB and, consequently, outcomes after cardiovascular surgery. PMID- 19681302 TI - Australian and New Zealand perfusion survey: management and procedure. AB - In this report, we will discuss management and procedural aspects of perfusion practice. This report allows us to compare and contrast recent trends and changes in perfusion with historic practices. A survey comprised of 233 single-answer and 12 open-ended questions was sent by e-mail to senior perfusionists or individuals in charge of perfusion in 40 hospital groups. The survey encompasses a review of the perfusion practices for the calendar year of 2003, and respondents were required to answer the survey based on the predominant practice in their institutions. Standard management of routine adult cardiopulmonary bypass (CPB) in 2003 consisted of perfusion strategies that achieved a target temperature of 32.0 degrees C (range, 28.0-35.0 degrees C), a flow index of 2.4 L/min/m2 (range, 1.6-3.0 L/min/m2) during normothermia and 1.8 L/min/m2 (range, 1.2-3.0 L/min/m2) during hypothermia, and a pressure during CPB between 50 (range, 30-65 mmHg) and 70 mmHg (range, 60-95 mmHg). Myocardial protection with blood cardioplegia was used in 77% of the 20,688 CPB cases, whereas in 53% cases, cardiotomy blood was never processed. Pre-operatively, 76% of perfusion groups assessed their patients (21% directly with the patient), and 85% responded that perfusionists performed or participated in a formal pre-bypass checklist. The majority of the perfusion groups used a handwritten perfusion record (62%), 12% used an electronic perfusion record, and 26% used both, whereas more than one half of the groups were involved in quality assurance (79%), incident reporting (74%), audits (62%), research (53%), participating in interdisciplinary meetings (53%), and morbidity and mortality meetings (65%). Only 26% conducted formal perfusion team meetings. This report outlines the status of clinical management and procedural performance for perfusion practices in Australia and New Zealand in 2003. Awareness of these trends will allow perfusionists to assess both individual practices and unit performance. PMID- 19681303 TI - Trends and emerging technologies in extracorporeal life support: results of the 2006 ECLS survey. AB - Extracorporeal life support (ECLS) is a procedure used to support the failing heart and/or lungs via a heart lung machine. Over 145 institutions perform this practice in the United States with more than 24,000 ECLS cases recorded. While many articles are published each year on common perfusion practice, little information is shared on emerging technologies in ECLS and common practices among perfusionists and ECLS specialists. This article presents our 2006 ECLS survey results and discusses emerging technologies and management topics new to the ECLS arena. ECLS specialists were asked to participate in an online survey. Two hundred twenty-two ECLS specialists responded. This survey suggests positive displacement roller pumps are still the leading pump used for ECLS 122/188 (64.9%). Silicone membrane oxygenators are used by responders 75% of the time for long-term use, while hollow fiber membrane oxygenators are used 44%. Forty-five percent of responders are using heparin or biocoated circuits exclusively, while 14.6% restrict their use to specific subpopulations. The most common coating is heparin coating (67.9%). Activated clotting time (ACT) management is still standard of care for coagulation monitoring (98%), while partial thromboplastin time (PTT) follows at 71.7%. The interquartile range for ACTs is 160-220 seconds and 160-200 seconds with active bleeding. This article suggests ECLS specialists are beginning to incorporate different technology into their practice, such as centrifugal pumps with hollow fiber oxygenators and coated-circuits. PMID- 19681305 TI - Invited commentary: be prepared to change your mind. PMID- 19681304 TI - Anti-inflammatory effect of aprotinin: a meta-analysis. AB - It is important to define the extent, and any limitations, of potential anti inflammatory regimens used in cardiac surgery to guide the rational combination of drugs to suppress the systemic inflammatory response. Aprotinin (Trasylol) is an anti-fibrinolytic agent with reported anti-inflammatory properties. In this study, we investigated the published data on aprotinin's effect on acute phase protein and cytokine levels in cardiac surgery patients. Randomized placebo controlled trials of aprotinin published between 1985 and 2007, in adult cardiac surgery using cardiopulmonary bypass, reporting tumor necrosis factor-alpha (TNF alpha), interleukin-6 (IL-6), IL-8, and IL-10 levels were included for review. Two independent reviewers graded each paper and collected information on inflammatory markers. RevMan 4.3 statistical software was used to calculate and plot the weighted mean difference between placebo and aprotinin groups. Thirteen studies met the review criteria. None of the inflammatory markers were reduced by high-dose aprotinin treatment. Low-dose aprotinin significantly reduced IL-10 levels after protamine administration (-41.3 pg/ mL; 95% CI: -59.5, -23.1), but this result was gone by the first post-operative day. These meta-analyses showed no significant effect of aprotinin on acute phase proteins or systemic cytokine markers of inflammation during clinical adult cardiac surgery using cardiopulmonary bypass. While recognizing that other host defense systems, such as coagulation and complement, contribute to the overall systemic inflammatory response, the evidence presented here does not support the clinical use of aprotinin as an anti-inflammatory agent on its own. PMID- 19681306 TI - Gravity separation of pericardial fat in cardiotomy suction blood: an in vitro model. AB - Fat emboli generated during cardiac surgery have been shown to cause neurologic complications in patients postoperatively. Cardiotomy suction has been known to be a large generator of emboli. This study will examine the efficacy of a separation technique in which the cardiotomy suction blood is stored in a cardiotomy reservoir for various time intervals to allow spontaneous separation of fat from blood by density. Soybean oil was added to heparinized porcine blood to simulate the blood of a patient with hypertriglyceridemia (> 150 mg/dL). Roller pump suction was used to transfer the room temperature blood into the cardiotomy reservoir. Blood was removed from the reservoir in 200-mL aliquots at 0, 15, 30 45, and 60 minutes. Samples were taken at each interval and centrifuged to facilitate further separation of liquid fat. Fat content in each sample was determined by a point-of-care triglyceride analyzer. Three trials were conducted for a total of 30 samples. The 0-minute group was considered a baseline and was compared to the other four times. Fat concentration was reduced significantly in the 45- and 60-minute groups compared to the 0-, 15-, and 30-minute groups (p < .05). Gravity separation of cardiotomy suction blood is effective; however, it may require retention of blood for more time than is clinically acceptable during a routing coronary artery bypass graft surgery. PMID- 19681307 TI - Carotid artery diameter, plaque morphology, and hematocrit, in addition to percentage stenosis, predict reduced cerebral perfusion pressure during cardiopulmonary bypass: a mathematical model. AB - Cerebral complications after cardiac surgery are a significant cause of morbidity, mortality, and financial cost. Numerous risk factors have been proposed to explain the risk of cerebral damage. Carotid artery disease has an important role. Percentage carotid artery stenosis is the only measure of carotid artery disease that is used by cardiac surgeons to determine the need for either a carotid endarterectomy and/or a higher pump perfusion pressure. Identification of patients through their carotid plaque morphology who might benefit from higher pump perfusion pressures or concomitant carotid endarterectomy may reduce cerebral morbidity and mortality. A mathematical model using finite element analysis was created to model the carotid artery vessel and its stenotic plaque. Analysis showed that the degree of carotid artery stenosis, the length of the carotid artery plaque, the diameter of the carotid artery, and the blood hematocrit all independently significantly affect the required pump perfusion pressure to maintain adequate cerebral perfusion during cardiopulmonary bypass (CPB). The results from a mathematical model showed that carotid artery diameter, carotid artery plaque length, and hematocrit, in addition to percentage stenosis, should be included in any thought process involving carotid artery stenosis and cardiac surgery. Estimating cerebral risk during CPB should no longer rely on only the percentage stenosis. PMID- 19681308 TI - Influence of cardiopulmonary bypass on the interaction of recombinant factor VIIa with activated platelets. AB - Recombinant factor VIIa (rFVIIa) interacts preferentially with coated platelets characterized by a high exposure of phosphatidyl serine (PS), FV, FVIII, FIX, and FX binding, and fibrinogen. Cardiopulmonary bypass (CPB) is known to impair platelet function. In this study, the influence of CPB on formation of coated platelets and the interaction of rFVIIa with the platelets were studied. Blood was either exposed to a closed CPB circuit or obtained from patients undergoing CPB-assisted cardiac surgery, and platelets were analyzed by flow cytometry with and without dual agonist stimulation with thrombin and a GPVI collagen receptor agonist known to induce coated platelet formation. Platelets circulated within a closed CPB circuit did not spontaneously form coated platelets. Dual agonists stimulation caused formation of coated platelets at a reduced level compared to pre-CPB level (51 +/- 21% vs. 80 +/- 17% before CPB, p < .001). The rFVIIa interaction with the coated platelets was not impaired after CPB. Platelets isolated from patients undergoing CPB-assisted cardiac surgery also formed coated platelets only after dual agonist stimulation but to the same level as before surgery (76 +/- 8% vs. 83 +/- 14% before surgery, p = .17, n = 10). rFVIIa interaction with the coated platelets was not impaired after surgery. No spontaneous rFVIIa-binding platelets were found. The data indicate that CPB exposure in vivo does not compromise the platelet-dependent effects of rFVIIa either by spontaneous formation of coated platelets, thereby limiting the risk of systemic coagulation, or by impairing rFVIIa interaction with the agonist-induced coated platelets, thereby retaining the hemostatic potential of rFVIIa after CPB. PMID- 19681309 TI - Triboelectric charging and dissipation characteristics of the Maquet Quadrox-D membrane oxygenator. AB - Triboelectric charging is commonly detected during cardiopulmonary bypass in circuits using roller pumps and PVC tubing. Dissipation of this charging is needed to prevent a spontaneous discharge from occurring. We evaluated the ability of the Quadrox-D (Maquet) to effectively remove the electrostatic charge accumulation, with and without a heater/cooler connection (H/C). A Quadrox-D oxygenator was evaluated using a Stockert SIII pump head, Cincinnati subzero H/E, and a custom adult extracorporeal membrane oxygenation tubing pack with 1/2" PVC raceway. The circuit was primed and evaluated for triboelectric accumulation with and without H/C use. The results showed a linear relationship between increasing pump flow and static charge buildup when an H/C was not applied. The calculated r2 value was .95. Incorporation of the H/C effectively eliminated charge accumulation. Increasing pump speed increases the amount of static charge created without the use of an H/C. Incorporation of an H/C effectively eliminates charge build-up in the Quadrox-D and is recommended while priming the circuit. PMID- 19681310 TI - Massive carbon dioxide gas embolism: a near catastrophic situation averted by use of cardiopulmonary bypass. AB - Endovascular vein harvesting is used as a less invasive method when compared to the standard open surgical method of dissecting and procurement of the greater saphenous vein. The benefits include smaller incision, decreased blood loss, less pain, decreased transfusion, decreased risk of infection and enhanced epidermal cosmetics. However, endovascular vein harvesting does have a learning curve to master the technique and although endovascular vein harvesting is the preferred standard of practice over the open surgical technique, it does have inherent potential complications. During endovascular vein harvesting, there is a potential for the patient's circulatory system to collapse which can be identified initially through hemodynamic monitoring, blood gas results, and vigilance through clinical observation. The suspected source of the crisis was confirmed when the surgeon cannulated the right atrium, where a massive gas source that (appeared pressurized) escaped upon incision of the right atrial appendage. Cardiopulmonary bypass (CPB) was utilized to support the patient and rectify the impending catastrophic event. Once full CPB was attained, we achieved hemodynamic stability and eventually all blood gases were normalized. Massive CO2 embolism is a life threatening emergency which must be identified and corrected instantaneously. CPB was the modality used to salvage this situation. Attention to the set-up of the EVH equipment, use of the transesophageal echocardiography, cerebral cximetry, vigilance and cooperation of all disciplines in the OR are definite recommendations to prevent such an occurrence. Experience gained by the perfusion team with a previous case was applied and helped to solve the immediate problems presented in this case. PMID- 19681311 TI - Treatment of malperfusion during surgery for type A aortic dissection. AB - During surgery for acute type A aortic dissection, malperfusion may occur during cardiopulmonary bypass. Retrograde perfusion trough femoral cannulation is considered a predisposing factor. However, this may occur even with antegrade perfusion, because of the presence of multiple flaps or compression of the true lumen by the false lumen. In this particular setting, the aim is to reach a perfusion of the true lumen of the dissected aorta. A technique of epicardial ultrasound-guided direct cannulation of the dissected aorta's true lumen by a Seldinger technique may help in those cases. We describe the technique with particular interest to the epicardial ultrasound control and the type of arterial cannula to be inserted by the Seldinger technique. PMID- 19681312 TI - Should tourniquets be used in upper limb surgery? A systematic review and meta analysis. AB - This study compares the intra- and post-operative outcomes of upper limb orthopaedic surgical procedures performed with and without tourniquet assistance. A systematic review was undertaken assessing the electronic databases Medline, CINAHL, AMED and EMBASE. The evidence-base was critically appraised using the Cochrane Bone, Joint and Muscle Trauma Group quality assessment tool. Study heterogeneity was statistically tested using Chi2 and I2 statistics. Where appropriate a random-effects meta-analysis was undertaken to pool results of primary studies assessing mean difference of each outcome. Two studies investigating fifty-five patients undergoing upper limb surgery were identified. The limited findings suggest that the use of tourniquets may reduce the incidence of technical difficulties during upper limb surgery. It remains unclear whether the application of a tourniquet can influence pain perception or operative duration. The evidence-base was considerably limited in both size and methodological quality. Further study is recommended to address the literature's methodological weaknesses. PMID- 19681313 TI - Comparison of early and delayed failed total shoulder arthroplasty. AB - The purpose of the present study is to determine what factors contribute to early failure after total shoulder arthroplasty (TSA). Implants were retrieved from 69 patients after failed TSA and were retrospectively assigned to two cohorts based on time of failure: Early, less than 2 years (N = 34); and Delayed, longer than 2 years (N = 35). The clinical information, intraoperative information, most recent radiographs, and damage mapping were collected for all patients. Patients in the Early failure group were significantly older (63.9 +/- 9.5 years) than those in the Delayed group (49.9 +/- 12.8 years) by an average of 14 years. The proportion of osteoarthritis cases was significantly higher for the Early group compared to the Delayed group (62% vs. 40%). The Delayed group had higher damage scores for several damage modalities. Elderly age and osteoarthritis were significant factors that were associated with early failures after TSA. PMID- 19681314 TI - Distal metaphyseal radius fractures in children: reduction with or without pinning. AB - We retrospectively reviewed 39 distal metaphyseal radius fractures in children. This study compares the results of closed reduction with or without percutaneous pinning. Twenty-four fractures were treated by closed reduction and above-elbow cast immobilisation. Re-displacement was noted after one week in three patients, for which they were revised with reduction and pinning. Fifteen patients were initially treated by closed reduction and pinning. In these patients we saw no redisplacement after six weeks immobilisation in a forearm cast. We recommend closed reduction and pinning as a predictable and safe alternative for unstable distal metaphyseal radius fractures. PMID- 19681315 TI - Discharging pin sites following K-wire fixation of distal radial fractures: a case for pin removal? AB - The aim of this study was to find out whether discharging pin sites following Kirschner-wire fixation of distal radial fractures warrant early wire removal. In a prospective study of 50 patients with a closed distal radial fracture treated by manipulation and either percutaneous or mini-incision Kirschner-wiring, we identified 14 patients (28%) with discharging pin sites. All of the cases were observed within the first two weeks following surgery. Of the 14 cases, only 2 patients had a positive swab culture and were treated with systemic antibiotics. In all cases the Kirschner wires were left in situ until serial radiographs demonstrated satisfactory union. Three patients (8.6%) in the percutaneous group had superficial radial nerve irritation, which settled following wire removal. Deep soft tissue infection, pyoarthrosis, osteomyelitis, and extensor tendon rupture were not encountered. We conclude that pin site discharge, whether sterile or infected, is in general not an indication for early wire removal. PMID- 19681316 TI - The Darrach procedure for post-traumatic reconstruction. AB - Over a 6-year-period, 15 women and 11 men with a mean age of 53 years (range, 24 to 80 years) had resection of the distal part of the ulna (Darrach's procedure) to address stiffness, instability, non-union, or substantial radioulnar length discrepancy after trauma. At an average follow-up of 21 months (range, 4 to 60 months), the improvement in total arc of forearm rotation averaged 87 degrees (range, 0 degrees to 160 degrees), from an average of 49 degrees to an average of 136 degrees (p < 0.001). The proportion of patients with occasional or continuous pain after the Darrach procedure (7 after vs. 16 prior ; p = 0.04) was significantly reduced. Only two patients had reoperation related to the residual ulna. In this study, the Darrach procedure improved forearm rotation and pain in patients with posttraumatic stiffness, instability, nonunion, or substantial radioulnar length discrepancy with a low complication and re-operation rate. PMID- 19681317 TI - Outcome of four-corner arthrodesis for advanced carpal collapse: circular plate versus traditional techniques. AB - Twenty eight patients with advanced carpal collapse (20 with scapho-lunate advanced collapse [SLAC] and 8 with scaphoid-nonunion advanced collapse [SNAC]) who were treated with a four-corner arthrodesis were reviewed; 23 had a good or excellent outcome. The mean DASH score was 41. The flexion/extension arc decreased from 72 degrees to 52 degrees. Gripping force increased from 46% to 72%. The range of motion was better in those with a traditional fixation (K-wires or screws) compared to the newer circular plates. There was no significant difference concerning pain relief, gripping force and disability. PMID- 19681318 TI - Traumatic hip dislocation in children. AB - Traumatic hip dislocation is rare in children. The purpose of this study was to investigate the epidemiological features, dislocation types, treatments, and clinical and radiological outcomes. Seven cases of traumatic hip dislocation in children treated between 1996 and 2006 were included in this study. There were six boys and one girl with a mean age of 6.5 years. Six children had a low-energy injury. One child had a road traffic accident. All had a posterior dislocation of the hip without any associated fracture. All children underwent closed reduction of their dislocation. The mean time interval between dislocation and reduction was 4 hours and 50 minutes. Following reduction, they were immobilised for six weeks: skin traction was applied for 3 weeks, followed in six children by a hip spica cast and in one child by non weight bearing mobilization. The mean follow up was 6.7 years. After clinical examination the hip was classified as normal in 6 children. One child had a stiff hip and a radiograph showed signs of avascular necrosis. The severity of injury was related to the age at the time of injury. Factors predisposing to avascular necrosis were delayed reduction and severity of trauma. PMID- 19681319 TI - Outcome of varus derotation closed wedge osteotomy in Perthes disease. AB - The aim of this study is to evaluate the clinical and radiological outcome of varus derotation osteotomy in Perthes disease. We studied 45 children with a mean age of 9.2 years, belonging to Herring's lateral pillar group B and C treated with varus derotation osteotomy. Containment was achieved in all patients on postoperative radiographs. At a mean follow-up of 6.4 years, good results were obtained in 23, fair in 20, and poor in 2 patients using Catterall's postoperative classification. Radiological evaluation was done using Mose's index and the epiphyseal quotient. Statistical analysis has concluded that better outcome was achieved in patients younger than 10 years of age as compared to those older than 10 years. Our study suggests that varus derotation osteotomy is an effective and easy surgical containment method for children with severe Perthes disease, especially who are younger than 10 years of age. PMID- 19681320 TI - Long-stem revision prosthesis for salvage of failed fixation of extracapsular proximal femoral fractures. AB - A retrospective review was made of radiographs and case notes of patients with failed fixation of extracapsular proximal femoral fractures subsequently managed with long-stem revision arthroplasty. Follow-up radiographs, objective scoring, mobility, and complications were assessed. Twenty five hips were managed with long-stem hip arthroplasty in 24 patients with a mean age of 73 years. The mean follow-up was 24 months. Patients received uncemented acetabular components and long-stem uncemented femoral implants. Complications included two intraoperative femoral fractures which were strut-grafted, three wound infections (one required washout), and one recurrent dislocation managed conservatively. Average postoperative Oxford Hip score was 29. We report a low complication rate and no specific implant related problems, with good functional outcome as evidenced by the outcome scores and mobility status following salvage arthroplasty. PMID- 19681321 TI - Impact of stem-broach sizing on the cement mantle of Lubinus SP II stems. A CT scan analysis. AB - Lubinus SP II stems are cemented either line-to-line with the largest broach or one-size undersized. The purpose of this study was to compare both implantation techniques. We used 18 polymeric stem replicas cemented line-to-line and undersized in paired cadaveric femora and analyzed them with CT scan images. Cementing Lubinus stems line-to-line resulted in higher medullary canal-filling indices (28.26 +/- 4.10%), thinner cement mantles (3.29 +/- 0.40 mm), more cement defects (5.12 +/- 1.69%) and more areas of thin cement (23.81 +/- 7.13%) than undersizing (respectively: 23.61 +/- 4.24%, 3.62 +/- 0.43 mm, 1.48 +/- 2.04%, 15.11 +/- 5.93%). In both settings, over 80% of areas of thin or deficient cement were supported by cortex. Using a line-to-line technique, adequate stem alignment was achieved without distal centralizer. Undersizing the stem and using a distal centralizer reduced the incidence of distal cement defects by a factor 10. While stems cemented line-to-line might have mechanical advantages, undersizing and using distal centralizers reduced potential pathways for debris migration to the bone-cement interface. PMID- 19681323 TI - High tibial valgus osteotomy using the Tomofix plate--medium-term results in young patients. AB - Proximal tibial valgus osteotomy is one of the treatment options for painful medial compartment osteoarthrosis and varus deformity of the knee in a young patient. We report our experience with medial opening wedge tibial osteotomy using the Tomofix plate in 46 patients (50 knees). The mean age was 39.5 years (range 30-49). All were male. Mean duration of follow-up was 60 months (36-72 months). There were no non-unions of the osteotomy site and the medial open-wedge healed without any need for bone graft or bone substitutes. There was functional improvement, as seen from the Oxford and Knee Society scores. Preoperative average knee flexion was 110 degrees (range, 90 - 125 degrees) which remained unchanged. The mean preoperative tibio-femoral angle (mechanical) was 7 degrees varus (range, 5-10 degrees); the postoperative angle was 6 degrees valgus (range 5 degrees-8 degrees). One knee was revised to total knee replacement after two years and was considered a failure. The Tomofix plate provided immediate stability, satisfactory healing of the osteotomy site without the need for bone graft or bone substitutes, and good functional results in young patients with isolated medial compartment degenerative disease. PMID- 19681322 TI - Rotatory stability of the knee after arthroscopic meniscus suture repair: a 5-to 17-year follow-up study of isolated medial and lateral meniscus tears. AB - The long-term rotatory stability of meniscus suture repair has not been firmly established clinically. Up to now there are only experimental studies done. This retrospective study aimed to compare the long-term rotatory stability of a knee with a meniscus suture repair with the stability of the uninjured knee in each of a cohort of patients. We evaluated both knees of 64 patients at an average follow up of 11 years (range : 5 to 17) after successful arthroscopic meniscus suture repair. Each patient's injury was an isolated longitudinal-vertical meniscus tear and each patient's opposite knee was uninjured. All repairs were performed with the same outside in meniscus suture repair technique. Evaluation included standardised clinical examination, anterior stability testing with a ligament testing device, and rotational testing with a rotational laxiometer. In the stability assessments, the mean anteroposterior translation was the same for repaired and uninjured knees: 3 mm at 67 N and 5 mm at 89 N. The mean external rotation was the same in repaired knees and uninjured knees at 20 degrees of flexion (22 degrees) and similar in repaired knees (22 degrees) and uninjured knees (23 degrees) at 90 degrees flexion. The mean internal rotation was similar in repaired knees (12 degrees) and uninjured knees (13 degrees) at 20 degrees and 90 degrees of flexion. Our findings indicate that a knee with arthroscopic meniscus suture repair displays a rotational stability that is equal to or nearly equal to the stability of an uninjured knee. However, whether rotational stability will only be preserved due to meniscus repair and how much of the meniscus needs to be preserved in order to maintain the biomechanical stability of the knee remains unclear. PMID- 19681324 TI - Bifocal and trifocal bone transport for failed limb reconstruction after tumour resection. AB - The aim of the present study is to assess the results of bifocal and trifocal bone transport for elimination of bone defects due to failed limb reconstruction after tumour resection. Thirteen patients, nine with giant-cell tumour and four with osteosarcoma, with bone defects resulting from wide tumour resection were managed by bone transport with an Ilizarov frame. The ages ranged from 12 to 46 years. Bone transport was used as a second line of management after failure of other modalities for reconstruction. According to the Enneking system for the functional evaluation after surgical treatment of musculoskeletal tumours, the percentage rating of function was 47% in one case, 70% in 2 cases and more than 85% in 10 cases. Most of the complications were treated successfully during the course of treatment. Bone transport with two or three osteotomies is a reliable method for eliminating bone defects as a limb salvage procedure after failure of other modalities for reconstruction. PMID- 19681325 TI - Complications following correction of the planovalgus foot in cerebral palsy by arthroereisis. AB - Pes valgus is a pathological condition which occurs in up to 25% of patients with cerebral palsy. Its correction in early age is essential to prevent progression of the deformity and to optimize the patient's function. In younger patients arthroereisis can be considered as a treatment that fills the void between orthotics and arthrodesis. We treated 15 patients (27 feet) with intra- or extra sinus tarsi arthroereisis. Concomitant Achilles tendon lengthening was performed in 12 feet. Results were good in 19 feet and poor in 8. Even though arthoereisis presents some complications, it can be considered a useful treatment to delay or avoid a Grice subtalar arthrodesis in flexible pes valgus due to cerebral palsy. PMID- 19681326 TI - Results of triple arthrodesis in children and adolescents. AB - We retrospectively evaluated 38 patients who had a total of 52 triple arthrodeses. The average age at the time of operation was 14.2 years and the mean followup period was 10.2 years. Results were rated good in 17 feet, fair in 28 and poor in 7. Talonavicular pseudarthrosis was observed in two feet, residual varus deformity in four, residual valgus deformity in one, wound infections in two, and wound haematomas in six. Better results were observed in cavovarus and planovalgus foot deformity, whereas poor results were especially seen in rigid equinovarus, calcaneoplanovalgus and equinoplanovalgus deformity. The role of supplementary operations performed prior to or concomitantly with the triple arthrodesis is crucial for correction of the deformity. Triple arthrodesis is a reliable procedure which generally produces good and satisfactory results. Very often, it needs to be supplemented by other operations for correction of complex foot deformities, as well as for the functional improvement of the patients. PMID- 19681327 TI - Long-term functional outcome after type A3 spinal fractures: operative versus non operative treatment. AB - The authors retrospectively studied, by questionnaires, the long-term (5 years) functional outcome after operative (posterior instrumentation: 38 cases) and non operative treatment (25 cases) for type A3 spinal fractures (Comprehensive Classification) without neurological deficit. A possible bias of this study was the fact that the operative group included 60% A3.2 and A3.3 fractures, versus only 12% in the nonoperative group. Two disease-specific questionnaires were used: the Visual Analogue Scale Spine Score and the Roland-Morris Disability Questionnaire. At follow-up the mean VAS scores were 82.6 and 80.8 in the operatively and non-operatively treated group, respectively; the difference was not significant. The mean RMDQ scores were 3.3 and 3.1 in the operatively and non operatively treated groups, respectively; again the difference was not significant. Functional outcome appeared to be equally good five years after operative or non-operative treatment of type A3 "burst" fractures. PMID- 19681328 TI - Does timing matter in performing kyphoplasty? Acute versus chronic compression fractures. AB - The objective of this prospective consecutive cohort study was to compare the clinical outcomes, the radiographic outcomes and the complication rates of symptomatic acute (< 10 weeks) and chronic (> 16 weeks) osteoporotic vertebral compression fractures (VCFs) treated with kyphoplasty. Twenty-eight consecutive patients had 52 symptomatic osteoporotic VCFs treated with kyphoplasty; 5 of these patients were treated in two sessions. The Oswestry Disability Index (ODI) for back pain, a Visual Analog Scale for pain assessment, a patients' satisfaction scale, and medication usage served to evaluate the clinical outcomes. All these variables improved significantly in both groups, and more so in the acute group, but the difference was most often not significant. Vertebral height, local kyphosis angle, global sagittal alignment and dynamic fracture mobility significantly improved in both groups (except global sagittal alignment), and again more so in the acute group (except global sagittal alignment); the difference between groups was significant regarding radiological variables, except global sagittal alignment. Timing of kyphoplasty certainly matters, as the clinical and radiological outcomes were mostly better in acute fractures than in chronic fractures, which somehow responded satisfactorily. Controlled studies (kyphoplasty versus natural history) are needed to establish the real value of the procedure. PMID- 19681330 TI - Fishtail deformity as a result of a non-displaced supracondylar fracture of the humerus. AB - Fishtail deformity is a very rare complication of undisplaced supracondylar fractures of the humerus in children. We report the case of a 10-year old girl presenting with pain in the right elbow eight years after a non-displaced supracondylar fracture of the humerus. Radiographs also demonstrated necrosis of the lateral part of the trochlea and of the head of the radius. With this long term clinical and radiographic follow-up after a non-displaced supracondylar fracture of the humerus, we would like to point out the possibility of the development of this very rare deformity. PMID- 19681329 TI - Dorsally angulated proximal phalanx fractures: closed reduction and rigid fixation using a reversed extension splint. AB - Dorsally angulated proximal phalanx fractures have always presented a difficulty for treatment. A variety of options for treatment have been tried in the past, many of these are operative options and therefore carry the risks of a surgical procedure. We present a case of a conservatively managed proximal phalanx fracture using a reversed dynamic or static finger extension splint, such as Roylan Sof-Stretch. The finger was immobilised using this splint and achieved bony union and very good function at both the metacarpophalangeal and proximal interphalangeal joints. PMID- 19681331 TI - Dislocation of the mobile bearing component of a patellofemoral arthroplasty: a report of two cases. AB - Isolated degenerative osteoarthritis of the patellofemoral joint can be treated with a patellofemoral arthroplasty. Improvements in patellar resurfacing designs have resulted in diminished complication rates. We describe two cases of dislocation of the polyethylene bearing, which is an unusual complication of a mobile bearing metal-backed patellar component of a patellofemoral arthroplasty. PMID- 19681332 TI - Augmentation of a patellar tendon repair with an autologous semitendinosus graft. AB - Patellar tendon ruptures are rare but very invalidating lesions. We describe a case of re-rupture of a sutured patellar tendon. An autologous semitendinosus graft was used for tendon augmentation in the reconstruction procedure. This technique uses an easy-to-harvest graft which has low donor site morbidity. Additionally, the strength of the graft allows early rehabilitation, and no further surgery for hardware removal is necessary. For these reasons we recommend this procedure for acute patellar tendon ruptures with a poor tissue quality or for revision surgery of the patellar tendon. PMID- 19681333 TI - Pseudodystrophy of the ankle and lower leg in a 9-year-old girl. AB - A 9-year-old girl presented with pain, swelling, redness and functional impairment of the left foot after a minor trauma. Clinical assessment revealed atrophy of the left calf and cyanosis and coldness of the left foot. Bone scintigraphy showed diffusely decreased tracer uptake in the left lower leg. Further examinations were normal. Pseudodystrophy was diagnosed and intensive physiotherapy was started. This resulted in complete functional recovery. Pseudodystrophy is typically found in children, adolescents and young women. The clinical features usually include severe pain at a joint or part of a limb with major functional disability, cyanosis, coldness and oedema or atrophy. This is caused by disuse of the affected limb after a minor trauma. The physical lesions may be improved or cured by means of intensive physiotherapy, sometimes combined with medication. As psychogenic factors often play an important role, one of the important elements of treatment is psychotherapy. The most important differential diagnosis is reflex sympathetic dystrophy (RSD). The distinction can be made by bone scintigraphy. PMID- 19681335 TI - A case of infantile scurvy treated only with vitamin C: a forgotten disease. AB - Scurvy is a rare disease occurring because of a nutritional deficiency of vitamin C. In the paediatric age group the disease is usually characterized by musculoskeletal manifestations. Treatment is straightforward and consists of vitamin C administration. However, if the patient is left untreated, scurvy may be life-threatening. We report here the case of a 16-month-old infant with scurvy. After proper treatment, the complaints disappeared in a very short time period and the boy grew up as a completely normal child during the 12 years follow-up. Nowadays only few physicians have experience with this disease, and ascorbic acid deficiency can thus easily be overlooked. With this paper we aimed to remind of the efficacy of vitamin C administration for patients with scurvy. PMID- 19681334 TI - Posterior epidural mass: can a posteriorly migrated lumbar disc fragment mimic tumour, haematoma or abscess? AB - A 60-year-old woman complained of low back pain radiating to both buttocks and to the anterior aspect of the left thigh. MRI showed a left posterolateral epidural mass at the L1-L2 level. An epidural abscess was suspected, but the biochemistry was normal. Excision yielded complete relief of symptoms. Pathological examination demonstrated that the specimen was a migrated disc fragment. The authors found 29 other cases of disc migration to the posterior epidural space; two of these were at the thoracic level. Eleven of the 27 lumbar cases (40%) were complicated with Cauda Equina Syndrome (CES). MRI is the method of choice to make the diagnosis. The differential diagnosis includes tumour, haematoma and abscess. PMID- 19681336 TI - Comparative effectiveness research: get to know the term. PMID- 19681337 TI - Pain control after cryotherapy. PMID- 19681338 TI - What is your diagnosis? Multiple dermatofibromas. PMID- 19681339 TI - Acute generalized exanthematous pustulosis: an enigmatic drug-induced reaction. AB - Acute generalized exanthematous pustulosis (AGEP) is a diffuse pustular disorder that is primarily drug induced and characterized by acute, extensive, small, nonfollicular, sterile pustules that usually begin in intertriginous folds with widespread edema and erythema. This article reports a case in which thalidomide, dexamethasone, or meloxicam may have been the etiologic agent to induce AGEP and the skin condition may have worsened with administration of additional medications during hospital admission. A good thorough medical history, including a drug history, along with clinicopathologic correlation is extremely important in a patient presenting with acute diffuse pustular lesions. PMID- 19681340 TI - A comparative split-face study of cryosurgery and trichloroacetic acid 100% peels in the treatment of HIV-associated disseminated facial molluscum contagiosum. AB - A comparative split-face study of 20 participants with disseminated facial molluscum contagiosum (MC) and human immunodeficiency virus (HIV) infection was conducted to assess treatment with cryosurgery (left side of the face) versus trichloroacetic acid (TCA) 100% peels (right side of the face). A total of 2 treatments were administered at 4-week intervals. At week 8, treatment with TCA 100% proved to be superior to cryosurgery with an average reduction in lesion count of 90% versus 55%, respectively. Demonstrating a statistically significant rate of lesion clearance (P < or = .05), TCA 100% appears to be more effective than cryosurgery at treating disseminated facial MC lesions in the setting of HIV infection. PMID- 19681341 TI - Ellis-van Creveld syndrome: case report and review of the literature. AB - Ellis-van Creveld syndrome (EVCS) is a rare. autosomal recessive disorder. We present a 9-year-old boy who was referred to the dermatology clinic for evaluation of congenital nevi. His history was consistent with the classic tetrad of EVCS. We discuss this potentially serious condition with congenital heart malformations that can result in failure to thrive and even death if not recognized early. PMID- 19681342 TI - Necrolytic acral erythema: a review of the literature. AB - Necrolytic acral erythema (NAE) has been described as an early cutaneous marker for hepatitis C virus (HCV) infection. It most commonly presents as a well defined, dusky, erythematous eruption with marked hyperkeratosis and a dark red rim associated with pruritus or burning. Necrolytic acral erythema bears microscopic and clinical resemblance to other necrolytic erythemas, including necrolytic migratory erythema (NME) and several nutrient-deficient syndromes. It is distinct, however, in its predominantly acral distribution and strong association with HCV infection. The pathogenesis is unknown, but a relationship to metabolic alterations has been hypothesized. Optimal therapy appears to be treatment of the underlying HCV infection using a combination of ribavirin and interferon alfa; oral zinc therapy may be an alternative but useful therapy. Cases of NAE without HCV infection suggest that more work needs to be done defining NAE and its relationship to HCV. PMID- 19681343 TI - Exercise-induced vasculitis associated with autoimmune disease. AB - Exercised-induced vasculitis (EIV) is an underreported and frequently misdiagnosed condition that occurs on the lower extremities shortly after exercise. Most reported cases have presented in healthy-appearing individuals, but some cases have been linked to other disease processes. A case report is presented of recurring EIV in a 65-year-old woman with a history of dermatitis herpetiformis; chronic, mildly elevated liver transaminases of unknown cause; microscopic colitis; celiac disease; multiple miscarriages; and heart block who was found to have autoimmune hepatitis upon workup of her rash. Both EIV and autoimmune hepatitis were misdiagnosed over many years by several clinicians in various specialties. Her family history was remarkable for 2 sisters with systemic lupus erythematosus and similar recurring exercise-induced rashes of the lower extremities, suggesting a familial link for this condition. Clinicians should recognize EIV and consider the possibility that this disorder may be the presenting sign of subclinical connective-tissue diseases. PMID- 19681344 TI - Study of trolamine-containing topical emulsion for wound healing after shave biopsy. AB - The efficacy and safety of trolamine-containing topical emulsion (Biafine) for enhancing wound healing after shave biopsy were evaluated in a single-center, open-label, single-group study. Fifteen participants applied trolamine-containing topical emulsion twice daily to the entire wound area for 4 weeks. Twelve of 15 participants (80%) completed the study. Wound healing was evaluated at weeks 1, 2, and 4. After 1 week of treatment with trolamine-containing topical emulsion, wound size was reduced by half. Complete healing occurred by week 4. Intention-to treat (ITT) analysis of the investigators global assessment (IGA) of efficacy (primary end point) over time showed trolamine-containing topical emulsion was moderately to very effective in 80% of participants (12/15) at week 1, 86.7% (13/15) at week 2, and 80% (12/15) at week 4; corresponding figures for the perprotocol analysis were 85.7% (12/14), 100% (13/13), and 100% (12/12), respectively. Treatment was well-tolerated with, mild or moderate application site signs (ie, erythema, erosion, inflammation, crusting) present during the first 2 weeks of treatment; mild or moderate subjective symptoms (ie, irritation, itchiness, burning, tenderness, pain) were present predominantly during the first 2 weeks of treatment. One participant withdrew from the study because of application-site erythema/burning probably related to trolamine-containing topical emulsion. These data indicate that 4 weeks of treatment with trolamine containing topical emulsion promotes rapid healing and is safe and well-tolerated in patients who undergo shave biopsies. PMID- 19681345 TI - Safe and effective treatment of seborrheic dermatitis. AB - Seborrheic dermatitis is a common chronic inflammatory skin disorder that can vary in presentation from mild dandruff to dense, diffuse, adherent scale. The disorder occurs throughout the world without racial or geographic predominance; it is more common in males than females. Its precise etiology remains unknown, but the condition is strongly associated with lipophilic Malassezia yeasts found among the normal skin flora and represents a cofactor linked to several risk factors, including T-cell depression, increased sebum levels, and activation of the alternative complement pathway. The goal of treatment is symptom control, with an emphasis on the importance of maintaining patient adherence to therapy to achieve low rates of recurrence. Available therapies include corticosteroids, antifungal agents, immunomodulators, and medicated keratolytic shampoos. Although corticosteroids are associated with recurrence, they sometimes may be recommended in combination with antifungal agents. Antifungal therapy is considered primary, but some agents are more effective than others because of their favorable pharmacokinetic profiles, high rates of absorption, anti-inflammatory and antipruritic properties, and vehicle. PMID- 19681346 TI - [Lasers: principles, characteristics and tissue interactions]. AB - Since their appearance in 1960, lasers have been considered useful light sources for medical applications. Laser light is monochromatic, the bundle is parallel and can be directed and focussed, as a result of which very high energy densities can be achieved. Several applications in dentistry have been investigated over the past decades. This article describes the physics behind lasers, the characteristics of the laser beam and overviews the laser wavelengths currently used in dentistry. The different interactions between the laser beam and the target are explained. PMID- 19681347 TI - [Lasers in general dental practice, an added value]. AB - Laser treatment in promoting dental care is present in many areas and disciplines. Modern practice management implies also the introduction of new technology. As there is evidence of the added value for lasers in different disciplines in dentistry practitioners should not be hold back and not wait for patients demanding for this technology for dental treatment. PMID- 19681348 TI - [Cavity preparation using an Er:YAG laser in the adult dentition]. AB - Many lasers (i.e., different wavelengths) are available today for clinical applications. Not all lasers are to be used for cavity preparation and removal of carious tissues. Erbiumlasers (Er:YAG and Er,Cr:YSGG) are suitable for these purposes (the wavelengths coincide with the highest absorption peaks of water and hydroxyapatite). The advantages during cavity preparation and caries removal are smear layer free cavity walls, selective and localised removal of tooth substance, a restricted need or the absence of anaesthesia, and cavity walls with a higher acid resistance resulting in a better protection against secondary decay. Possible side effects of erbium lasers with water cooling and used with respect for correct power setting are minimal and can be compared with those of rotary instruments. Erbium laser tooth preparation is now one of the options in the minimal invasive approach. The needs for acid etching has been debated for long. At present it has become clear that acid etching is mandatory to obtain a good adhesion and retention with resin composites. PMID- 19681349 TI - [Comparative study on dogs between CO2 laser and conventional technique in direct pulp capping]. AB - Three dogs have been used in this experiment. Class V cavities were made in sixty teeth. A pulpal communication was provoked intentionally in these cavities. Teeth were randomly split in 2 groups (30 teeth for each). On first group, the pulp bleeding was stopped until appearance of coagulum on exposed pulp surfaces by means of CO2 laser irradiation (Output Power: 3 W, Pulse duration: 0.1 sec, frequency: 1 Hz, spot size diameter: 0.3 mm, Energy density: 425 J/cm2). Calcium Dihydroxide was deposited followed by a temporary filling (IRM, Dentsply, De Trey, Germany). In the second group, the calcium Dihydroxide was deposited directly on exposed bleed pulp (conventional technique) followed by the same temporary filling. Ten weeks later, all teeth were extracted and prepared for histological study. RESULTS: 93% of treated teeth preserved their pulp vitality in the group treated with CO2 laser for direct pulp capping versus 82% in the group treated by conventional technique. The average of the thickness of the dentinal bridge newly formed was 391.5 +/- 33 microm for teeth irradiated with laser and 294.1 +/- 28 microm for teeth treated by conventional technique. The thickness of the dentinal bridge newly formed in teeth treated by means of CO2 laser was 33% more important than in those treated by the conventional technique. Statistical analysis showed a significant difference between the averages of values in all groups (p < 0.05). To conclude, CO2 Laser use in the direct pulp capping increases significantly the percentage of pulp vitality preservation and the thickness of the dentinal bridge newly formed after pulp exposition. PMID- 19681350 TI - [Surgical treatment of maxillary osteonecrosis due to bisphosphonates using an Er:YAG (2940 nm) laser. Discussion of 17 clinical cases]. AB - Reports of cases of ONJ are significantly increased during the last five years as a iatrogenic complication of therapy with bisphosphonates (BPT). The aim of this work is to present the advantages of surgery using Er:YAG laser for treatment of ONJ. Er:YAG laser can gradually reach the healthy bone without causing any heating damage of tissues. This device results very versatile and gives the possibility of choose among different surgical techniques depending by the case (e.g.: vaporization or ostectomy). Moreover, different studies have demonstrated the presence of both bactericidal and biomodulating effect on bone and surrounding tissues, with biostimulation of microcirculation and neoangiogenesis. Seventeen sites of ONJ, classified according to the staging system developed by Ruggiero and observed in 12 patients with multiple myeloma (9 patients), bone metastases (2 patients) and osteoporosis (1 patient), were treated with Er:YAG laser (Fidelis Plus, Fotona-Slovenia). Laser device was used in non-contact or near-contact way (VSP, 300 m3 30 Hz, Fluence 60 J/cm2) on 17 sites (4 Stage I and 13 Stage II) on 3 different types of surgery: sequestrectomy + debridement, sequestrectomy + corticotomy and vaporization. For an average follow-up of 9 months (SD +/- 6 months), complete healing of ONJ (Stage 0) was obtained for 13 sites (76.5%) and resolution of symptoms was obtained (Stage 1) for 3 sites (17.5%). For one site at Stage II (6%), recovery was obtained but this result was not maintained over 3 months. Positive results were independent by the anatomical area (mandible or maxilla), primary disease (osteoporosis, multiple myelomas or metastasis) and discontinuation of BPT before surgery. Er:YAG laser (2940 nm), in our experience, represents a valid therapeutic option for ONJ-BP related, especially in early stages of the disease. PMID- 19681351 TI - [Original work. The impact of tobacco use on the oral cavity. 1946]. PMID- 19681352 TI - [A critical and ironic instant in science and attitude of health personnel]. AB - Smoking habits have a negative impact on general health and are detrimendous for teeth, dental implants, periodontal and peri-implant tissues. In this paper, however, a critical note is placed based on a comparison of the current evidence based knowledge with scientifically sound communications dating 1946. It seems that not only science has turned its own conclusions upside down but also health care workers and dentists have changed their opinion regarding smokers enormously. Some reflection and relativism should be taken into account when dentists are planning dental care in smoking patients. PMID- 19681353 TI - Interview with Mark R. Neaman, FACHE, president and chief executive officer, NorthShore University HealthSystem. Interview by Kyle L. Grazier. PMID- 19681354 TI - The lifelong iterative process of physician retention. PMID- 19681355 TI - Helping hands: leaders in the healthcare sustainability movement. PMID- 19681356 TI - Organ transplantation: is the best approach a legalized market or altruism? AB - More than 100,000 individuals are on the waiting list to receive a lifesaving transplant, but many of them will not receive the organ they need. Legislation has been passed to support organ donation--the current organ procurement system in the United States is governed by the Uniform Anatomical Gift Act of 1968 and the National Organ Transplant Act of 1984--but a shortage of donated organs remains. Potential solutions include a legalized organ market, expanded legislation, therapeutic cloning, and xenotransplantation. If no change occurs, the black market in organ procurement will continue to expand. A comprehensive approach consisting of both short-term (increased education and national paired kidney exchange) and long-term solutions (expanded legislation to fund and promote therapeutic cloning) is presented in this essay. PMID- 19681357 TI - Consumer-driven healthcare marketing: using the web to get up close and personal. AB - This essay examines the emergence of consumer-driven healthcare marketing, including its operational definition, how it has been used in the past, and how it has evolved. Specifically, marketing practices in other industries are inspected to understand the factors that have contributed to their successes and to determine the relevance of these efforts to healthcare marketing. The advantages of new, technology-enabled marketing opportunities are considered as well, such as stealth ads, blogs, podcasts, and corporate participation in social networks. The implications of the regulation on healthcare websites, along with the work-around strategies used, are analyzed. Lastly, the essay submits recommendations for the healthcare executive when implementing a consumer-driven healthcare marketing plan. PMID- 19681358 TI - Factors associated with hospital bankruptcies: a political and economic framework. AB - Between 2000 and 2006, 42 U.S. acute care hospitals filed for bankruptcy protection under federal law. This article explores hospital bankruptcies over a six-year period. Bankrupt hospitals are compared with their competitors, and hospitals surviving bankruptcy are compared with those organizations that eventually close. Finally, this article identifies nonfinancial factors associated with the filings and categorizes these factors into a political and economic framework. A literature review of hospital trade publications is used to identify organizations filing for bankruptcy during this period. Data gathered from these resources are used in concert with American Hospital Association data to identify hospital characteristics and publicly available information on factors surrounding hospital bankruptcy filings. Data on the status of hospitals after filing are also collected to determine whether bankruptcy reorganization is successful or results in hospital closure. Results indicate that 67 percent of hospitals filing for bankruptcy during this time eventually ceased operating. Bankrupt hospitals are smaller than their competitors. They are also less likely to belong to a system and more likely to be investor owned. Factors associated with filing organizations are placed into a political and economic framework derived from Park's work on municipal bankruptcy filings. Common nonfinancial factors associated with hospital bankruptcies include mismanagement, increased competition, and reimbursement changes. PMID- 19681359 TI - New Hampshire critical access hospitals: CEOs' report on ethical challenges. AB - Research into the importance of organizational healthcare ethics has increasingly appeared in healthcare publications. However, to date, few published studies have examined ethical issues from the perspective of healthcare executives, and no empirical study has addressed organizational ethics with an explicit focus on rural hospitals. For our study, we sought to identify the frequency of ethical conflicts occurring within 12 general categories (domains) of administrative activities. Also, we wanted to determine what ethics resources are currently available and whether additional resources would be helpful. We conducted a structured telephone interview of all 13 chief executive officers (CEOs) of critical access hospitals in New Hampshire. All the CEOs in the study indicated that they encountered ethical conflicts. On average, the three most frequently noted domains were organizational-professional staff relations, reimbursement, and clinical care. All CEOs indicated they would like to have additional ethics resources to address these conflicts. This study verified that CEOs encounter a broad spectrum of ethical conflicts and need additional ethics resources to address them. Because this study used a small sample of CEOs and represented only one New England state, further ethics-related research in rural healthcare facilities is warranted. Follow-up study would allow for (1) a higher level of generalization of the findings, (2) clarity regarding specific ethical dilemmas that rural healthcare executives encounter, and (3) an assessment of ethics resources and training that healthcare executives need to address the ethical conflicts. PMID- 19681360 TI - [Teenage pregnancy as a risk factor for fetal abuse]. AB - OBJECTIVE: To identify if pregnancy in adolescence is a risk factor for fetal abuse. MATERIAL AND METHOD: A case-control study of 333 mothers that made fetal abuse and its controls, was made between October 2005 and May 2006 at the Nuevo Hospital Civil de Guadalajara Dr. Juan I. Menchaca. The fetal abuse was considered when the prenatal attention was deficient and was documented illicit drugs or physics violence to the mother. The fetal abuse was documented by means of a direct interview to the mother. Was realized a descriptive analyze of different forms of fetal abuse. The odds ratio was calculated for association between adolescent mother and fetal abuse. RESULTS: The prenatal attention was deficient in 284 (69.7%), illicit drugs abuse in 100 (24.5%) and physics violence to the mother en 69 (20.7%). The adolescent mothers had more deficient prenatal attention that adult mothers (54 vs. 37.7%), used more illicit drugs (25.6 vs. 16.6%) and received more physics violence (4.9 vs. 2.8%). The pregnancy in adolescent was associated with fetal abuse OR: 1.9 (95% CI: 1.39-2.71). CONCLUSIONS: The pregnancy in adolescent is a risk factor to fetal abuse and the form more frequent was the deficient prenatal attention. PMID- 19681362 TI - [Expression of muscarinic cholinergic receptors in human term placenta]. AB - BACKGROUND: Studies in the human term placenta have shown that muscarinic cholinergic receptors M1-M4 are located in the sincitiotrofoblasto in the basement membrane and in the brush border membrane of the villi. However, in both studies the methodology has not been implicated microscopy methods to reveal its location. OBJECTIVE: To determine the expression of muscarinic cholinergic receptors M1 and M2 in human term placenta. MATERIALS AND METHODS: We analyzed 20 samples of human placenta at term newborns. Muscarinic cholinergic receptors M1 and M2 were detected by immunohistochemistry and polyclonal antisera. The optical density of the signal for each muscarinic cholinergic receptor was analyzed with Image Pro Plus software. 30 readings were made by the placenta (n=600). The expression of muscarinic cholinergic receptors were analyzed using the Student t test. RESULTS: The expression of muscarinic cholinergic receptors M1 and M2 in human term placenta were found in the sincitiotrofoblasto of secondary and tertiary villi. The expression of muscarinic cholinergic receptors M1 and M2 in the population showed that the RCM M2 is expressed in a greater proportion than RCM M1 (p < 0.01). The expression of muscarinic cholinergic receptors M1 and M2 was lower in newborns with low weight for gestational age, although the difference was not significant (p > 0.05). CONCLUSIONS: The expression of muscarinic cholinergic receptors were identified in placental sincitiotrofoblasto predominantly M2 muscarinic cholinergic receptor. The values reported here represent a baseline that can be used to analyze the expression of muscarinic cholinergic receptors in the placenta of women with a history of environmental exposure to toxic substances. PMID- 19681361 TI - [Correlation index amylase-creatinine clearance to endogenous creatinine clearance in severe preeclampsia]. AB - INTRODUCTION: Tubular lesion may cause acute renal insufficiency in pregnant patients with severe preeclampsia. OBJECTIVE: To describe the correlation between the amylase/creatinine clearance ratio and endogenous creatinine depuration in pregnant patients with severe preeclampsia. MATERIAL AND METHOD: Transversal study (pilot study) twenty eight women with pregnancies of 20 to 40 weeks complicated by severe preeclampsia were studied. Subjects had serum and urine creatinine and amylase determinations to calculate the amylase/creatinine clearance ratio (%). According to the results, two groups were formed: group A (> 3%) and group B (< or = 3%). The correlation between amylase/creatinine clearance ratio and endogenous creatinine depuration was evaluated. STATISTICAL ANALYSIS: measures of central tendency and dispersion, Student's t-test, Pearson correlation coefficient (r) and linear regression were used. RESULTS: Group A included 23 cases (82%) and group B included 5 cases (18%). Amylase/creatinine clearance ratio (%) for group A was 5.22 +/- 1.6 and for group B was 2.41 +/- 0.41 (p = 0.001). The endogenous creatinine depuration (mL /min. /1.73 m2 SC) for group A was 105.6 +/- 9.71 and for group B was 132.10 +/- 7.95 (p = 0.54). The r between amylase/creatinine clearance ratio and endogenous creatinine depuration for group A was -0.43 and for group B was -0.25. CONCLUSIONS: A moderately significant negative correlation exists between amylase/creatinine clearance ratio and endogenous creatinine depuration. PMID- 19681363 TI - [Vaginal adenosis and vaginal intraepithelial neoplasia: a review of the literature and a case report]. AB - Vaginal adenosis is an uncommon pathology, characterized by the presence of cylindrical epithelium in the vagina. This pathology is considered rare if not found in women exposed in utero to diethylstilbestrol (dietilestilbestrol). Vaginal intraepithelial neoplasia (VAIN) is another uncommon condition in women who have not been hysterectomized to treat cervical intraepithelial neoplasia (CIN), with an incidence of 0.3 per 100,000 women. On the other hand 0.9% of the patients who have been poshysterectomized to treat CIN develop VAIN. This paper aims at reviewing the literature on vaginal adenosis, vaginal intraepithelial neoplasia and their possible treatments. In addition, the paper reports on a case of vaginal adenosis in a young female patient with concomitant VAIN 2, the CO2 laser ablation performed, and the short-term results obtained. PMID- 19681364 TI - [Hyaline membrane in the death of the newborn in cesarean section. 1954]. PMID- 19681365 TI - [Clinical practice guidelines 2008. Mexican College of Obstetrics and Gynecology Specialists. Management of preterm premature rupture of membranes]. PMID- 19681366 TI - [Marking techniques and ultrasound guided mastografia prior to obtaining biopsy breast excisional]. AB - BACKGROUND: Breast cancer is the most common gynecologic malignancy worldwide. Localization under ultrasound guidance or mammography prior to biopsy is one of the procedures used for diagnosis of subclinical malignant or premalignant lesions. OBJECTIVE: To describe techniques for localization by mammography or ultrasound at our institution, prior to excisional breast biopsy. MATERIAL AND METHODS: Localization under ultrasound guidance or mammography from July 1 to December 31, 2008 at the HospitalAngeles del Pedregal. An analysis of the category BIRADS, histopathologic results, the imaging procedure and the surgical approach. RESULTS: We made 78 procedures. 56% under mammography guidance and 44% by ultrasound. 23% of biopsies with histopathological result of malignancy and 77% were benign, the outcome depended on the category of BIRADS granted. Success depends on the technique of covering the lesion in its entirety and was 100% in our study. CONCLUSIONS: Localization of nonpalpable lesions of the breast remains a useful procedure for diagnosis of suspicious lesions. The choice of the imaging technique depends on the characteristics of the lesion under study. PMID- 19681367 TI - [Body composition in women with gestational diabetes mellitus]. AB - BACKGROUND: Several techniques have been used to determine body composition during pregnancy. OBJECTIVE: To determine the characteristics of body composition in women with gestational diabetes mellitus in comparison with women with normal glucose tolerance and pre-gestacional diabetes. MATERIAL AND METHODS: Pregnant women with gestational diabetes mellitus, pre-gestacional diabetes, and normal glucose tolerance, between 24 to 32 weeks of single gestation, were enrolled in a cross-sectional study. Screening of DMG was carried out using 50 g of glucose load; diagnosis was confirmed by oral glucose tolerance test. Evaluation of body composition was carried out by bioelectrical impedance. The Kruskal Wallis test was used for statistical analysis. RESULTS: A total of 79 women were included; of these, diagnosis of gestational diabetes mellitus, pre-gestacional diabetes, and normal glucose tolerance was established in 14, 9, and 56 women, respectively. Pre-gestational body mass index was greater in women with diabetes (p < 0.01). Fat free mass and total body water were similar in the studied groups. Fat mass was greater in women with gestational diabetes mellitus (range 21.0-29.4 kg) and patients with pre-gestacional diabetes (range 26.4-32.7 kg) than in the women with normal glucose tolerance (range 150.8-25.9 kg), p < 0.01. CONCLUSION: The body composition of women, between 24 to 32 weeks of single gestation, is different in the women with gestational diabetes mellitus compared with women with normal glucose tolerance. Women with gestational diabetes mellitus show a significant increase in fat mass without significant changes in the fat free mass and total body water. PMID- 19681368 TI - [Confounding effect of prematurity in the neonatal death associated with obstetric maternal morbidity]. AB - BACKGROUND: Prematurity is considered the main factor of neonatal mortality in developed countries (60 to 80% of cases). OBJECTIVE: To determine if obstetric morbidity and/or prematurity are associated with neonatal death. PATIENTS AND METHODS: A cohort of 25,365 live newborns since January 1st 2000 to December 31st 2004 was studied. Neonatal mortality was compared according to the number of prenatal visits, single or multifetal pregnancy, the presence or not of preeclampsia/eclampsia, cesarean section or vaginal delivery, and duration of rupture of membranes, stratifying by weeks of gestational age or by preterm and term gestation, as it was convenient. Chi-square test and Odds Ratio (OR) with 95% Confidence Intervals were calculated (CI). RESULTS: There was not significant statistical difference in neonatal mortality at less number of prenatal visits, between single and twin pregnancies, in the presence of preeclampsia/eclampsia and pregnancies without complications, when they were stratified by group of gestational age. When it was controlled gestational age, malformations and maternal-fetal and obstetrical morbidity, there was not difference in mortality of neonates born vaginally or by cesarean section. It was observed an increased risk or neonatal mortality in preterm neonates with 48 hours or more of rupture of membranes (OR 3.05 CI 95% 1.64-5.66) CONCLUSIONS: Performing and stratified analysis, prematurity was the factor associated with neonatal mortality, and not the number of prenatal visits, multifetal pregnancy, preeclampsia/eclampsia, or cesarean section. The duration of rupture of membranes is an independent factor of prematurity for neonatal mortality. PMID- 19681369 TI - [Tuberculosis of the breast. A case report]. AB - Breast tuberculosis is a rare form of tuberculosis, in developed nations its prevalence is 0.1%. In undeveloped nations it is between 3 and 4%. Less frequently breast tuberculosis cases are presented. Some texts deals with simple mastectomy cases understood as breast cancer (6, 8, 9). There is a case. She is a 35 years old woman; she is from Piedras Clavadas, Tihuatlan. She belongs to high prevalence tuberculosis community, and she has not received BCG vaccine. She presents painful tumor which has developed during 3 months in her left breast, and armpit adenomegaly. The ultrasonography report presents back breast abscesses and low response to antibiotic treatment. Chronic granulomatous mastitis with focal cheesy necrosis and isoleted Langhans' giant cells responding positively to antituberculosis treatment are reported by the biopsy. PMID- 19681370 TI - [Fusion of the labia minora as a cause of urinary incontinence in a postmenopausal woman. A case report and literature review]. AB - Labial fusion is rarely presented in the postmenopausal age group, its predisposing factors are hipoestrogenism and a inflamation/repair process in the vaginal epithelium. Our case report involves a 73 years old woman that presents with urinary incontinence. On physical exploration, complete labial fusion was noted, a surgical approach was undertaken with liberation of the labia and fixation, postoperatory local estrogen ointment was indicated with complete recovery. The literature review confirmed that this was the ideal treatment and also confirms the role of local estrogen ointment as profilactic and therapeutic. PMID- 19681371 TI - [Systemic lymphoma cells with T precursor condition of extreme female genital tract. A case report and literature review]. AB - Primary female genital tract non Hodgkin's lymphoma is a rare presentation for a common disease in the childhood, and its classification as primary extranodal lymphoma is still controversial. There are a few cases reported as a primary precursor B-cell lymphoblastic lymphoma of the female genital tract, but there is not any case reported as primary precursor T-cell lymphoblastic lymphoma of the ovary in childhood. Herein we describe a 16 years old young woman with bilateral ovarian tumors, paraaortic lymphoadenophaty and disseminate disease to the female genital tract including extension of the tumor to neighboring organs like the omentum and the appendix. Exploratory laparatomy were performed with bilateral salpingo-oophorectomy, hysterectomy, omentectomy, appendectomy, pelvic and para aortic lymphadenectomy, pelvic washings and with biopsy of vaginal vault. The chemotherapy regimen comprised of CHOP (Cyclophosphamide, Hydroxydaunorubicin, Oncovin, Prednisone/Prednisolone) and methotrexate, 3 months later presents left facial hemiparesia follow by right facial hemiparesia, 7 months later presents more Central Nervous System (CNS) complications and apparently was complicated with acute lymphocitic leukemia and after 16 months from the diagnosis, following by a torpid evolution, the pacient finally died. PMID- 19681372 TI - [Breast malignancies and their study in the gynecological pathology. 1954]. PMID- 19681373 TI - [Clinical practice guidelines 2008. Mexican College of Obstetrics and Gynecology Specialists. Uterine cervical cancer and pregnancy]. PMID- 19681374 TI - Making environmental literacy a foundation for nursing advocacy. PMID- 19681376 TI - Lost in the shuffle: culture of homeless adolescents. AB - Estimates indicate that approximately 1.7 million youth are homeless in the United States. Many associated risk factors have been identified for adolescent homelessness, including family conflict, leaving foster care, running away or being thrown away, physical or sexual abuse, and coming out to parents as gay, lesbian, bisexual, transgender, or questioning one's sexual identity (GLBTQ). The purpose of this ethnographic study was to explore the culture of homelessness for adolescents. Nineteen homeless adolescents from a major urban area in the northeast U.S. were observed and interviewed over an 18-month period. The elements of the street culture of homeless adolescents were identified by study participants' stories. For many study participants, the decision to live on the streets was a logical and rational alternative to remaining in possibly dangerous and unstable home environments. It provided a means to their generating social capital. Nevertheless, it can be concluded that existing programs and policies relative to adolescents who are at risk for homelessness or already living on the streets should be re-examined and redesigned to meet the unique needs of vulnerable youth so they do not get lost in the shuffle. PMID- 19681375 TI - Health-related quality of life in two itinerant samples: carnival and migrant farm worker children. AB - The document Healthy People 2010 sets a national health care agenda that includes reducing health disparities and improving quality of life. This study evaluated health-related quality of life (HRQOL) in children aged 2 to 12 years being raised in two itinerant populations: traveling carnival children (n=33) and migrant farm worker children (n=48), and compared their outcomes to each other and to findings in published literature. The study sample utilized cluster sampling from outdoor amusement companies (carnivals) and agricultural farms who agreed for the researcher to enter their premises and speak with their workers. The PedsQL Generic Core Scales, including a child self-report and parent-proxy, measured HRQOL. HRQOL of the itinerant children did not differ from that of a more geographically stable California sample. The carnival children's mean scores were higher than the migrant farm worker children's scores on 7 out of 12 subscales, but the differences were not statistically significant. PMID- 19681377 TI - There's a fine line...adolescent dating violence and prevention. AB - Domestic violence is a paramount issue in society today. One component of this is the violence associated with dating and interpersonal relationships in the teen years. Current research demonstrates that a significant percentage of teens have experienced some degree of physical, sexual, or emotional abuse from a dating partner. Pediatric nurses may interface with teens before, during, and after violent episodes. It is important for nurses to know the definitions of dating violence and rape, data about the incidence and prevalence of this issue, risk factors for violence in intimate relationships, dating violence myths, and the potential impact of youth dating violence. Youth perceptions and current research must be analyzed to frame current and potential interventions to reduce teen dating violence. Pediatric nurses have an integral role in counseling teens, ensuring youth safety, and preventing violence in the adolescent years. PMID- 19681378 TI - Dietary choices of urban minority high school students. AB - The purpose of this study was to gather data on the knowledge of nutrition and health, and the dietary choices of minority high school students in an urban setting. Focused group interviews, survey questionnaires, and food diaries were used to obtain information about the food choices made on a daily basis by culturally diverse high school students. The impact of the school meal program, as well as ethnic and cultural patterns, were strong influences in terms of the teens' dietary intake. This study offers insight into the limitations and obstacles to achieving a healthy diet for this population of students. PMID- 19681379 TI - Catch-up growth for the extremely low birth weight infant. AB - PURPOSE: This retrospective chart review describes the growth of extremely low birth weight (ELBW) infants after discharge from the neonatal intensive care unit. DESIGN: A descriptive design was used to collect data from 60 hospital records. RESULTS: Gains were noted between assessment periods (discharge--6 months and 6-12 months) (t = 4.57, t = 5.60, t = 10.77, p < 0.001) for weight, length, and head circumference, respectively. A negative change in weight (z = 1.9; CI = -3.1, -0.66) within the discharge to 6-month assessment period and a positive change in length (z = 0.54, CI = 0.05, 1.04) within the 6 to 12-month assessment were noted. Each assessment was significantly below the 50th percentile of full-term infants with weight at the 7th percentile at 12 months. DISCUSSION: The ELBW infants showed gains relative to the full-term infant but lagged behind on each growth parameter at each assessment. PMID- 19681381 TI - Why can't she speak? PMID- 19681380 TI - Research evidence review and appraisal: pediatric central venous catheter care bundling. AB - Central venous catheters are integral to the care of acutely ill children, providing reliable vascular access for infusions, hemodynamic monitoring, and blood sampling. However, there are risks associated with their use, the most common of which is central line-associated blood stream infections. These infections result in increased lengths of stay, increased costs, and high mortality rates. A thorough review of research evidence has been completed to fully appreciate the state of the evidence regarding the effects of bundling together the care for central venous catheters, and practice recommendations have been provided. Published studies have been appraised and evaluated for clinical and statistical significance. This appraisal has resulted in clear and specific recommendations for evidence-based practice applications, and potential policy implications are outlined in this article. PMID- 19681382 TI - Public health and drought. PMID- 19681383 TI - Responding out loud to budget disaster: the case of El Paso County, Colorado. PMID- 19681384 TI - Predictability of swimming prohibitions by observational parameters: a proactive public health policy, Stamford, Connecticut, 1989-2004. AB - Using compiled bacterial analyses to predict water quality when certain conditions are observed provides a way to establish a proactive public health policy. In this study, the authors reviewed using the geometric mean various conditions including the amount of rain in previous days, wind direction and speed, tides and high tide height, water temperature, drought or flood conditions for the season, different materials coming into the swimming areas, and the location and amount of any sewage spills. Only three events showed statistical significance (Chi-squared p < .0001): rain events of one inch or more in a 24 hour period under normal weather conditions; rain events over a half inch in a 24 hour period under drought conditions; and when "floatable" material from distant sewage spills (i.e., grease balls) are present at a beach. Such evaluations enable a public health policy to be easily developed that restricts swimming when certain adverse conditions are present without waiting for bacteriological examinations to prove that a problem exists. PMID- 19681385 TI - Mold growth on gypsum wallboard--a summary of three techniques. AB - Reducing occupant exposure to mold growing on damp gypsum wallboard and controlling mold contamination in the indoor environment was studied through 1) delineation of environmental conditions required to promote and avoid mold growth and 2) efficacy testing of antimicrobial products, specifically cleaners and paints, on gypsum wallboard (GWB) surfaces. The effects of moisture and relative humidity (RH) on mold growth and transport are important in avoiding and eliminating problems. These effects have been demonstrated on GWB and are discussed in this article for use as control guidance. The authors discuss the efficacy of antimicrobial cleaners and paints to remove, eliminate, or control mold growth on GWB. Research to control Stachybotrys chartarum growth using 13 separate antimicrobial cleaners and nine varieties of antimicrobial paint on contaminated GWB was performed in laboratory testing. GWB surfaces were subjected to high RH. GWB control measures are summarized and combined, and the antimicrobial product results are explained. PMID- 19681386 TI - Association of self-reported leisure-time physical inactivity with particulate matter 2.5 air pollution. AB - This study examines the association between annual levels of particulate matter (PM) and self-reported leisure-time physical inactivity (LTPI) in the Behavioral Risk Factor Surveillance System (BRFSS) among 63,290 survey respondents who participated in the 2001 BRFSS from 142 counties in the U.S. The average prevalence of self-reported LTPI was about 24.9% (SE = 0.3%), LTPI prevalence was positively associated with annual mean of PM.5 concentration (p < .0001). The authors demonstrate that LTPI was associated with PM2.5 pollution with statistical significance with and without adjustment for covariates (adjusted odds ration [OR] = 1.16; 95% CI: [confidence interval] 1.06-1.27). This study suggests that ambient PM2.5 air pollution is associated independently with LTPI. PM2.5 pollution and physical inactivity are both risk factors of chronic diseases. Therefore, it is important for environmental officials to implement measures to reduce ambient air pollution while public health officials simultaneously promote regular physical activity by encouraging the general public to remain physically active. PMID- 19681387 TI - The health impact of nonhazardous solid waste disposal in a community: the case of the Mare Chicose landfill in mauritius. AB - The disposal of nonhazardous solid waste on the island of Mauritius is centered on the Mare Chicose landfill. Because of health concerns in the community, the authors conducted a health impact survey, took measurements of height and weight, and compared the results with a control locality. After adjusting for confounding factors, the authors observed that numerous complaints were significantly higher in the target population. After elimination of reporting bias, however, nausea and vomiting remained as the only significant self-reported outcomes. The mean body mass index (BMI) of target men was significantly lower when compared with the control. This result was not observed in the populations of women and children, and suggests a gender difference in the health impact of the landfill. PMID- 19681388 TI - Responses to the efficacy of 'green' cleaning products article. PMID- 19681389 TI - Responses to the efficacy of 'green' cleaning products article. PMID- 19681390 TI - Responses to the efficacy of 'green' cleaning products article. PMID- 19681391 TI - Response to third-party review of cleaning products article. PMID- 19681393 TI - Representing your best interests! PMID- 19681392 TI - The U.S. Environmental Protection Agency's Pesticide Worker Safety Program- implementation of the National Strategies for Health Care Providers: Pesticides Initiative. PMID- 19681394 TI - [German poultry farming between animal welfare and global market]. AB - Despite the positive tendencies concerning sales output in the poultry production, the margins per single animal are extremely low. This circumstance leads inevitably to an increasing number of animals per farm. Also the German egg production is currently confronted with a great challenge due to changes of the legislation of animal welfare in animal farming (German Tierschutz Nutztierhaltungs-Verordnung), the EU-zoonosis-regulation (2160/2003) and because of the avian influenza difficulties. In addition, the globalization has tightened the competitive conditions during production. Therefore, innovation potential and specialization are mandatory premises for the continuity within a free market economy. In all farming systems there has to be made a consideration between animal welfare, economy and ecology, whereas, based on animal welfare, the "ethical limit" has the utmost importance. It has to be accounted for the concept of fulfilment of demand and prevention of harm. The success of agricultural animal farming depends, last but not least, on a good and robust state of health of the live stock. The German consumer will have to accept that a high quality and high welfare poultry product will have their price, even in the global market. The sale orientation on non-European production methods is not acceptable under the aspect of animal welfare. PMID- 19681395 TI - Comparative study on the behaviour, health and productivity of laying hens in a furnished cage and an aviary system. AB - A total of 540 Lohmann Silver laying hens was used to investigate behaviour, use of the structural elements offered, as well as health and productive parameters in six units (45 hens each) of one furnished cage system (FC; Typ 715/725, Fa. Salmet, Berge, Germany, according to EU guideline 1999/74/EG [2003]) vs. three units (90 hens each) of one aviary housing system (AV; type natura, Big Dutchman, Vechta, Germany). There was no difference between the two systems in terms of animal health, egg quality, laying performance and nest acceptance Acceptance of the nests was high in both systems and resulted in relatively small proportions of mislaid eggs (median 0.28% AV vs. 0.33% FC). Perches in FC were used during the light phase by 15.6% of the hens compared with 73.3% at night. The dust bathing activity of the hens in AV corresponded to natural circadian behaviour patterns (P <0.003) and the complete behaviour chain was exhibited. In contrast, FC hens invariably exhibited incomplete dust bathing patterns on the mat and on the grid floor without any diurnal rhythm. Duration of dust bathing of hens in FC differed significantly from the AV hens (median value 4.77 minutes on mats and 4.62 on grid floors respectively vs. 14.87 minutes, P < 0.05). Even though comparison of laying performance, egg quality and health did not yield any serious disadvantage of one system over the other, it has to be taken into consideration that normal behaviour was highly restricted in FC. PMID- 19681396 TI - [Measurement of floor space allowance of Lohmann Silver hens using biometric data]. AB - Space allowance for laying hens is one of the most disputed criteria of modern laying hen husbandry. Fast and cheap methods to achieve reliable data about the space occupied by laying hens are missing. This studie is based on a method, that can be used to measure the space ocupied by laying hens using digital images from above. 100 hens were photographed digitally when walking or scratching freely on a blue coloured wooden plate that had been placed in a sheltered outdoor scratching area. The birds were 65 weeks old and in the 47th laying week. The body width and the floor area covered by the hens were measured from the photographs by the KobaPlan color contrast planimetric method using a personal computer. Another 50 hens were randomly caught and weighed in order to get an estimate of the average bird weight of the herd. The mean floor space covered by the hens was 545.4 cm2 (SD: +/- 51.9) and their mean width was 173.9 mm (SD: +/- 8.9). The mean weight of the 50 hens was 2085.8 g (SD: +/- 234.0 g). Compared to the space allowance in the German "Kleingruppenhaltung" (small colony cage holdings) Lohman Silver laying hens occupy more than half (68.2% resp. 60.6%) of the minimum floor space of 800 cm2 per hen resp. 900 cm2 for hens heavier than 2 kg (Nutztierhaltungsverordnung 2006). Subsequently less than half of the floor space (32.6% resp. 39.8%) can be used for movement or other normal behaviours including social behaviour. Compared to the floor space of 750 cm2 per hen as laid down in the European Council Directive 1999/74/EC, 72.5% of the floor space in furnished cages is covered by the birds. Further data on the floor space covered by the body size of layers in different situations and from different genetic strains can be gained using the KoBaPlan method in order to estimate reasonable space allowances for laying hens in modern husbandry systems which in particular should enable the hens to perform normal behaviour. This is evenly important for hens which start laying with less than 2 kg but exceed 2 kg while in production as given in the German regulation. PMID- 19681397 TI - Bone strength and keel bone status of two layer strains kept in small group housing systems with different perch configurations and group sizes. AB - The objective of the present study was to investigate whether an arrangement of perches at two different heights within individual compartments of small group systems (back perch elevated (BE), front perch elevated (FE) or both perches heightened (FBE)) combined with an enlarged group size would increase humerus and tibia bone breaking strength and impact keel bone status. Bone strength and keel bone status of two layer strains (LSL, LB) kept in small group systems (SG 40-60 (40, 60 hens), SG 20-30 (20, 30 hens) with different perch configurations) and furnished cages (FC, 10, 20 hens, perches in standard position) were compared in two trials. Investigations were carried out in the laying months 6 and 12, comprising a total of 576 hens. When all compartments of SG 40-60 had been incorporated with perches at two different heights, humerus and tibia bone strength in LSL layers significantly increased compared to FC, whereas keel bone status was negatively impacted. Within SG 40-60, BE and FE perches significantly increased humerus strength in LSL layers compared to FBE perches. LB layers had significantly higher bone strength in groups of 20 compared to 30 hens in SG 20 30, whereas no effect of group size was detected for LSL hens. Keeping hens in SG 40-60 with modified perch positions was associated with increased bone strength but brought about the problem of inferior keel bone status. PMID- 19681398 TI - [Effect of litter type on prevalence and severity of pododermatitis in male broilers]. AB - During the course of two trial periods on broilers (Ross 308), the effects of 5 different types of litter (chopped straw, wood shavings, spelt glumes, Pelletinos, HygieneWood-Shavings) on the development of pododermatitis and animal performance (body weight, feed conversion, mortality) were investigated. Pelletinos are for sale under the name Pelletino Strohstreugranulat G. HygieneWood-Shavings are a special product that currently is not available on the market. Male broilers were kept under practical conditions (feeding, light program) for 35 days. Foot pad lesions could already be observed at the first assessment on their 7th day after hatching. Broilers kept on chopped straw showed in both trials the highest score (trial 1: 1.50; trial 2: 1.58). Animals reared on Pelletinos (trial 1: 0.59; trial 2: 0.17) followed by HygieneWood-Shavings (trial 1: 0.63; trial 2: 0.47), wood shavings (trial 1: 1.30; trial 2: 0.71) and spelt glumes (trial 1: 1.13, trial 2:0.93) showed significant lower lesions. During the second trial, the average body weights were significantly higher while feed conversion was lower.The mortality on the other hand showed no significant differences between the trials. Groups kept on Pelletinos showed the best foot pads and the highest body weights in both trials at the end of experiments. The investigation showed that the prevalence and the severity of pododermatitis as well as the performance could be influenced by various types of litter. Chopped straw which is standard for the broiler industry in Germany, seems to be the least suitable variant in our study. PMID- 19681399 TI - [Effect of locomotor activity on leg disorder in fattening chicken]. AB - Leg disorders have become a major concern in comercial poutltry meat production. The mechanisms which cause leg problems are not completely understood. It has been hypothesed that high growth rate and low locomotor activity are the main factors. The particular effects of the different factors have been elucidated in special experiments. The locomotor activity of broilers with different growth rates has been observed in deep litter compartments with videotracking technique. The effects of locomotor activity were analysed through a training on treadmills in slow and fast growing broilers. Fast growing broilers showed a lower activity level from week one onwards and there was a pronounced drop in walking distances in the third week of age (21 to 8 meters per hour), while the slow growing broilers maintained a high activity level from 36 m throughout the fattening period. Systematic training of fast growing birds on treadmills increased the locomotion ability, the bone density and thickness and reduced the bending and twisting in leg bones. Increasing the distance between feeders and drinkers may be a method to increase locomotor activity under practical conditions and may be a method to reduce the incidence of leg disorders. Based on the results of previous experiments, a final test was carried out placing the feeder and drinker at two varied distances apart from each other (2 and 12 meters) and monitoring the effects on growth rate, locomotor activity, bone development and leg weakness in broilers. A total of 4800 male and female broilers (in groups of 600) were tested under virtually real conditions. The larger distance between food and water led to a three times increased level of locomotor activity. Weight development and food utilisation were not negatively influenced. Groups of birds, whose food and water supply were placed 12 metres apart, had significantly less cases of leg weakness and deaths due to leg disorders (2%). Increased walking activated the bone building processes of the tibia. This method is a practical way for broiler farmers to decrease leg weakness. PMID- 19681400 TI - [Examination of rearing standards and health status in turkeys in Germany]. AB - Within the framework of a cooperative research project, sponsored by the Federal Ministry of Food, Agriculture and Consumer Protection in Germany (BMELV), turkey health as well as numerous aspects of animal welfare in various intensive commercial farms with varying rearing forms were examined. For this purpose extensive documentation of flock management and health status information was conducted over the past two years at the farms. Additional information was ascertained from the carcasses at the slaughterhouse. The first results of this study, which look at flock management, will be presented here. Upon reviewing the questionnaires, it is clear that on one hand almost all farmers orientate there stocking rate upon the allowed limits, on the other hand, however, when considering national general requirements, which are very important parameters thatargely influence animal health (i. e. litter quality and care of sick/injured animals) farmer compliance lacks. Especially the litter quality, which, among other parameters, plays a very important role in food pad health, proved to be unsatisfactory. Unfortunately, no instruments are available that measure the litter quality, yet. The status of footpad health was a major problem in all the rearing systems. However, the prevalence of epithelial necrosis and deep skin lesions varied immensely among different flocks of the same age. PMID- 19681401 TI - [Turkey fattening under extensive rearing conditions--selected parameters concerning health, performance and behavior]. AB - Within two consecutive studies (study I and study II), each involving a summer (su) and a winter mast (wi), 36 male turkeys per B.U.T. Big 6 (BUT) and Kelly Bronze (KB) origin were set on two seperated areas in free range and fatted over a period of 20 (winter mast in study II) and 22 weeks respectively. Aim of both studies was to analyze the effects of extensive rearing conditions on these two turkey origins concerning health, performance and behavior and whether seasonal climatic differences had any additional influence. Besides the aim in study II was to detect how intensely environmental enrichment (plateau and perches) was accepted by the turkeys and if co-housing of BUT and KB turkeys was possible. The occurence of skin lesions in the breast area (breast blisters and/or breast buttons) was significantly influenced by season (su > wi) and origin (BUT > KB). Malposition of the hind extremities was found significantly more often in the BUT origin, whereas the X-leg position was the most common observed leg abnormality. The mortality rate remained beneath 6% during three of the four fattening periods. Due to their genetical determination, the BUT constantly gained higher weights than the KB (wi > su).The intensity of acceptance of the structural elements was influenced by season (su > wi), forage (ecological > commercial) and time of day (night > day).The results of the studies lead to the conclusion that turkeys of the BUT origin can certainly be fattened under extensive rearing conditions alongside the KB origin, while providing good performance. PMID- 19681403 TI - [Water provisions for Muscovy ducks--behaviour at duck showers and modified plasson drinkers]. AB - Feather pecking and cannibalism are serious problems in keeping Moscovy ducks. Prevention of feather pecking by regularly applied beak and claw trimming are increasingly criticised by the public. The recommendation of the Council of Europe (COE) for the keeping of Muscovy ducks in farming systems calls for environmental enrichment including water for preening and bathing after December 31,2010. A total of 126 female Muscovy ducks (not beak nor claw trimmed) from commercial breeding lines were kept for 63 resp. 70 days in four compartments with 15-16 ducks each during two production cycles. Two pens where equipped either with duck showers or open water facility (modified Plasson drinker). Water provisions were made available for the ducks four hours daily at working days from their fifth week of life until slaughter. Behaviour at the water provision was registered and analysed for the number of ducks being engaged with water (944 hours recordings over 59 days from four pens analysed in five-minute-intervals (11,540 observations). Additionally 858 feather preening bouts (five a day for each compartment) were analysed for the duration of feather preening behaviour at the water provision. From the fifth to the tenth week of life the mean percentage of animals of a pen was significantly higher at the open trough (trough: 8,3% (+/ 5,37); shower: 4.9% (+/-6.1), Mann-Whitney p <0,01) and feather preening took significantly longer (trough: 134.4 sec (+/-154.8); shower: 111.2 sec (+/-152.0), Mann-Whitney p <0.01) than at the showers. Bout duration and percentage of animals observed at both water provisions increased with age. Nonetheless only ten percent of the feather preening behaviour exceeded five minutes. Most animals made use of water in the first hour of the time period when water was provided. In the first weeks of water provision open water troughs were used more often and preening behaviour was longer. When given the choice, younger ducks preferred open drinkers to showers while older ducks showed a higher preference for the duck showers. In future it may be useful to elaborate whether a combination of open water troughs in the first few weeks of the fattening period followed by the provision of showers when the birds have completed their feather coverage can help to reduce the frequency of feather pecking and cannibalism. PMID- 19681402 TI - [Open water systems for species-appropriate housing of Peking ducks--effects on behaviour, feather quality and plugged up nostrils]. AB - At present in Germany Peking ducks in commercial housing are on ground with litter and nipple drinkers only and there is no access to open water. With this study we investigated if open water drinkers can affect behaviour and health of ducks positively. In this investigation ten fattening trials with 11 424 ducks, kept with straw, were analyzed. 25% of the compartment (32 m2) were covered with perforated grid, with one of the tested water systems (Nipple drinker, Sparkcup, Shower, Hollow and Round drinker and modified Round drinker) installed. Besides parts of the behaviour of ducks, animal health and hygiene and also economic factors were taken into account. In contrast to pens with nipple-drinkers only, the ducks in pens with open water troughs/showers had the opportunity to exhibit their natural drinking behaviour and water associated activities, such as straining and preening the plumage with water in accordance with the animal welfare requirements. In the free-choice pens, the open water systems were significantly preferred, and over a 24-hour period, the ducks in watering areas with open water troughs showed higher activity. Limiting the access to the open water systems (4, 6 and 8 hours) led to more intense use per time unit. Ducks with access to nipple drinkers only showed a significantly higher percentage of plugged up nostrils than animals from pens with open water drinkers (p < or = 0.05). Open water drinkers also had a positive impact on the plumage condition.The open water drinking systems were very well accepted by the animals and are, concerning behaviour and animal health, an improvement in housing systems of Peking ducks. PMID- 19681404 TI - Bluetongue control in Europe--new challenges and achievements. PMID- 19681405 TI - Direct blood pressure monitoring. PMID- 19681406 TI - Surveys, drug names, surgical airways and safety. PMID- 19681407 TI - Evidence-based evolution of the high stakes postgraduate intensive care examination in Australia and New Zealand. AB - The fellowship examination for intensive care medicine in Australia and New Zealand, first held in 1979, has undergone four major periods of development and change since inception. These periods are characterised as: 1. 1979 to 1996- initiation and establishment of the exam as a relevant and comprehensive assessment process for a new specialty. 2. 1997 to 2001--revision to increase breadth of coverage, increase reliability for a growing number of candidates and ensure that each candidate received the same exam: Expansion: to incorporate assessment of CanMEDS skills (including communication, procedures and professional qualities). Lengthening: to increase the number of exposures, to ensure reliability. Quarantining of candidates: to allow the provision of a similar exam for each candidate. 3. 2002 to 2006--increasing emphasis on examiner training, standard setting and increasing feedback to candidates to improve the educational experience and guide exam preparation. Blueprinting of questions to maintain validity. 4. 2008 onwards--logistic revision to ensure feasibility for a rapidly growing number of candidates and refinement to apply modem standard setting and quality control. The exam has been regarded as a 'tough but fair' assessment in its 30 years of existence and the committee overseeing its development has aimed to continually review the process to maintain those qualities as well as reliability, validity and feasibility. The increasing number of candidates has allowed accumulation of usable statistics but has tested the feasibility of running such a labour intensive exam. To date, there have been 800 presentations to the exam with 498 successful candidates. PMID- 19681408 TI - Mechanism of cardiac preconditioning with volatile anaesthetics. AB - In recent years, there has been increased interest in the mechanisms involved in anaesthetic-induced cardioprotection. It is not thoroughly understood how volatile anaesthetics protect the myocardium from ischaemia or reperfusion injury, but the overall mechanism is likely to be multifactorial. This review examines the recent experimental and clinical research underlying the cellular and molecular mechanisms involved in anaesthetic-induced preconditioning. A variety of intracellular signalling pathways have been implicated in the protective phenomenon. Ischaemic preconditioning and anaesthetic-induced preconditioning share similar molecular mechanisms, including activation of guanine nucleotide-binding proteins, triggering of second messenger pathways, activation of multiple kinases, mediation of nitric oxide formation and reactive oxygen species release, maintenance of intracellular and/or mitochondrial Ca2+ homeostasis and moderation of the opening of adenosine-triphosphate-sensitive potassium channels. A more thorough understanding of the multiple signalling steps and the ultimate cytoprotective mechanisms underlying anaesthetic-induced preconditioning may lead to improvements in the management of ischaemia and/or reperfusion injury. PMID- 19681409 TI - Analgesia after caesarean delivery. AB - As the number of women giving birth by caesarean increases throughout most of the developed world, so too is research into postoperative pain relief for these women. Like most other post-surgical populations, the new mother needs effective pain relief so that she can mobilise early but she also has the added responsibility of needing to care for her newborn baby. There is no 'gold standard' for post-caesarean pain management; the number of options is large and the choice of method is at least partly determined by drug availability, regional and individual preferences, resource limitations and financial considerations. Most methods rely on opioids, supplemented with anti-inflammatory analgesics, nerve blocks or other adjunctive techniques. The aim of this review is to detail commonly used opioid-based methods and to review the evidence supporting non opioid methods, when incorporated into a multimodal approach to post-caesarean pain management. Areas of promising research are also discussed. PMID- 19681410 TI - Subarachnoid morphine, bupivacaine and fentanyl as part of combined spinal epidural analgesia for low anterior resection. A prospective, randomised, double blind clinical trial. AB - This study was designed to compare the efficacy of subarachnoid morphine alone or in combination with bupivacaine and fentanyl for combined spinal-epidural analgesia in colorectal surgery. This is a prospective, randomised, double-blind clinical trial. Sixty patients undergoing low anterior resection were assigned to one of three groups: subarachnoid morphine, bupivacaine and fentanyl, subarachnoid morphine and bupivacaine or subarachnoid morphine only. Epidural catheter placement and subarachnoid injection were done via a combined spinal epidural Epistar needle at L2-3. The epidural catheter was used for scheduled intraoperative bupivacaine and intermittent postoperative bupivacaine and morphine administration. Intraoperative epidural bupivacaine, intraoperative intravenous fentanyl use, time to first analgesia request, postoperative visual analogue scale pain scores, tramadol requirements and side-effects were recorded for 72 hours. Postoperative analgesia was comparable in all groups. Intraoperative fentanyl and bupivacaine consumption was lowest in the morphine, bupivacaine and fentanyl group. Time to first analgesia request was longer in the morphine, bupivacaine and fentanyl compared to the morphine group (P = 0.009). Tramadol use was lower in the morphine and bupivacaine group compared to morphine, bupivacaine and fentanyl (P = 0.017) on postoperative day two. There were no significant adverse effects. All patients ambulated the morning after surgery. The addition of bupivacaine and fentanyl to subarachnoid morphine did not confer any advantage on postoperative visual analogue scale scores and tramadol use, but lowered the need for additional intraoperative intravenous fentanyl and epidural bupivacaine and prolonged the time to first analgesia request. PMID- 19681411 TI - Cardiac complications and mortality rates in diabetic patients following non cardiac surgery in an Australian teaching hospital. AB - This retrospective study of diabetic patients undergoing non-cardiac surgery has identified that a greater number of patients are at risk of cardiac complications and death in the perioperative period than had previously been suggested. As well as insulin-dependent diabetic patients and patients with elevated creatinine (> 178 micromol/l) as previously found, our study suggests that non-insulin dependent diabetic patients and patients with creatinine > 120 micromol/l are also at increased risk of cardiac complications and death following non-cardiac surgery. This increases by a factor of six those diabetic patients at risk of perioperative complications from non-cardiac surgery and also increases the number of patients with renal failure similarly at risk. The study confirms similar risks of cardiac complications and death to other recently published data and suggests ongoing comparisons will contribute to quality assurance activities in anaesthesia and surgery. PMID- 19681412 TI - Potency of mephentermine for prevention of post-spinal hypotension. AB - This study was designed to determine the minimum effective dose (ED50) of mephentermine for prevention of post-spinal hypotension in women undergoing elective caesarean section. Dixon's up-down method of sequential allocation was used to determine the patient's dose of vasopressor drug. In our previous study, mephentermine appeared to be much more potent than ephedrine, so the present study was conducted using a lower initial dose and smaller dose interval for this vasopressor. Following administration of spinal anaesthesia, a prophylactic infusion of mephentermine was started with 5 mg infused over a period of 30 minutes as the initial dose and a dose interval of 1 mg. The ED50 of mephentermine of 3.7 mg (95% confidence interval 2.4 to 5.7 mg) was much less than that of ephedrine, as calculated in our previous study using the same methodology. Using these values gives a potency ratio of ephedrine to mephentermine of 1:6.8 (95% confidence interval 6.0 to 7.5). PMID- 19681414 TI - Effect of remifentanil on tracheal intubation conditions and haemodynamics in children anaesthetised with sevoflurane and nitrous oxide. AB - This study examined the effect of remifentanil added to inhalation agents on intubating conditions in children (aged one to seven years). As is our routine, all patients were premedicated with 0.004 mg/kg glycopyrrolate intramuscularly 30 minutes before anaesthesia to prevent bradycardia and salivary secretion, and received intravenous 1 mg/kg ketamine prior to transfer into the operating room to facilitate separation of the child from the parents. Laryngoscopy was attempted after ventilation with 8% sevoflurane in 50% nitrous oxide and 50% oxygen for two minutes, and intravenous administration of either a placebo bolus of saline (Group S) or one of two doses of remifentanil, 1 microg/kg (Group R1) or 2 microg/kg (Group R2) given over 30 seconds, commenced at the beginning of the sevoflurane administration. Only 18 of 32 patients (56.3%) in Group S had acceptable intubating conditions compared with 31 of 32 patients (96.9%) in Group R1 and 32 of 32 patients (100%) in Group R2 (P < 0.001). Mean blood pressure was lower in Group R1 and R2 compared with Group S (P < 0.005). Our routine use of ketamine and glycopyrrolate may have influenced the relative lack of significant hypotension and bradycardia. Sevoflurane, nitrous oxide and remifentanil provided acceptable conditions for tracheal intubation in children and could be an acceptable alternative to intravenous induction and neuromuscular blocking agents. PMID- 19681413 TI - The effects of dexmedetomidine and fentanyl on emergence characteristics after adenoidectomy in children. AB - This randomised controlled study evaluated the effects of fentanyl and dexmedetomidine on emergence characteristics of children having adenoidectomy and anaesthetised with sevoflurane. Ninety children, two to seven years of age and ASA physical status I, were studied. Children were randomly assigned to one of three groups of 30 children, with the study intervention injection given intravenously after intubation. Children in Group F received fentanyl 2.5 microg x kg(-1), children in Group D received dexmedetomidine 0.5 microG x kg(-1) and children in Group C received saline solution. Anaesthesia was induced with 50% N2O and 8% sevoflurane in O2 by mask and atracurium 0.6 mg x kg(-1) was administered for tracheal intubation. All children received paracetamol 40 mg/kg rectally one hour preoperatively and dexamethasone 0.5 mg x kg(-1) intravenously. The time to extubation was shorter in Group D than Group F. The eye-opening time was longer in Group F (16.1 +/- 5.3 minutes) than in Groups C (12.0 +/- 4.2 minutes) and D (12.7 +/- 3.2 minutes). The proportion of pain-free children in early recovery was significantly higher in Groups D (47%) and F (43%) than Group C (13%) (P < 0.05). The proportion of children with agitation scores > 3 was lower in Groups D 17% (5/30) and F 13% (4/30) than in Group C 47% (14/30) (P < 0.05). Fentanyl 2.5 microg x kg(-1) and dexmedetomidine 0.5 microg x kg(-1) had similar haemodynamic effects and emergence characteristics. Fentanyl has been safely used in children for many years. Further studies of dexmedetomidine safety and its interaction with other anaesthetic agents are required before recommending its routine use during general anaesthesia in children. PMID- 19681415 TI - Preventing pain on injection of propofol: a comparison between peripheral ketamine pre-treatment and ketamine added to propofol. AB - The purpose of this study was to examine possible peripheral mechanisms for the reduction of propofol injection pain by the addition of ketamine. We hypothesised that pH changes associated with the addition of ketamine to propofol decrease propofol-induced pain on injection. We compared the efficacy of intravenous ketamine pretreatment under tourniquet with ketamine added to the propofol. In the pre-treatment group, patients received ketamine 10 mg in a total volume of 1.0 ml with 0.9% saline (n = 94; Group P) under tourniquet for 30 seconds before administration of propofol after release of the tourniquet. In the mixture group, propofol 9 ml was mixed with ketamine 10 mg in 0.9% NaCl 1.0 ml (n = 94, Group M). Pain was assessed with a four-point scale: 0 = no pain, 1 = mild pain, 2 = moderate pain, 3 = severe pain at the time of propofol injection. The pH of propofol, ketamine and a range of propofol-ketamine mixtures were also measured. Forty-eight patients (51%) in Group P complained of pain on injection compared with 28 patients (30%) in Group M (P = 0.005). The pH of the 1% propofol-ketamine mixture was 5.84 while 1% propofol had a pH of 7.86. Our results support pH changes as a more important cause for the decrease in propofol injection pain with the addition of ketamine to propofol than a peripheral effect of ketamine. PMID- 19681416 TI - Application of the Laboratory Risk Indicator in Necrotising Fasciitis (LRINEC) score to patients in a tropical tertiary referral centre. AB - The objective of this study was to assess the applicability and usefulness of the Laboratory Risk Indicator in Necrotising Fasciitis (LRINEC) score in patients from our institution. A retrospective analysis was undertaken of the case notes of all patients admitted to our facility between January 2002 and December 2005 with the admission diagnosis of necrotising fasciitis and the application of the LRINEC score upon the initial blood tests. The sensitivity, specificity and likelihood ratios were then calculated for patients with a LRINEC score of > or = 6 compared with the findings of a surgical biopsy. Twenty-eight patients were identified as having the admission diagnosis of necrotising fasciitis on the hospital database and were eligible for the study. Ten of these had biopsy-proven necrotising fasciitis. With a cut-off score of > or = 6, the LRINEC score had a sensitivity of 80%, specificity of 67%, a positive predictive value of 57% and a negative predictive value of 86% in distinguishing the patients with proven necrotising fasciitis from those with severe soft tissue infections. The likelihood ratio of a positive biopsy was 2.4 and the likelihood ratio of a negative biopsy was 0.3. A cut-off value of > or = 6 of the LRINEC score was not overly sensitive or specific, and the likelihood ratios suggest that, at this cut off level, the LRINEC score would have only a very small effect on the post-test probability of the patients in the studied population having necrotising fasciitis. PMID- 19681417 TI - Effect of pre-emptive alveolar recruitment strategy before pneumoperitoneum on arterial oxygenation during laparoscopic hysterectomy. AB - In a randomised, controlled, single-blind trial, we examined the effect of a pre emptive alveolar recruitment strategy on arterial oxygenation during subsequent pneumoperitoneum. After intubation, 50 patients were randomly allocated to receive either tidal volume 10 ml/kg with no positive end-expiratory pressure (group C) or alveolar recruitment strategy of 10 manual breaths with peak inspiratory pressure of 40 cmH2O plus positive end-expiratory pressure of 15 cmH2O before gas insufflation (group P). During pneumoperitoneum, group P was ventilated with the same setting as group C (FiO2,= 0.35, tidal volume 10 ml/kg). PaO2, measured during peumoperitoneum was higher in group P than in group C (166 +/- 32 mmHg vs 145 +/- 34 mmHg at 15 minutes, P = 0.028, 155 +/- 30 mmHg vs 136 +/- 32 mmHg at 30 minutes, P = 0.035). Alveolar-arterial oxygen gradient in group P increased less after gas insufflation (13 +/- 9 to 60 +/- 34 mmHg vs 10 +/- 9 to 37 +/- 31 mmHg, P = 0.013). We conclude that the alveolar recruitment strategy we applied before insufflation of the peritoneal cavity may improve oxygenation during laparoscopic hysterectomy PMID- 19681418 TI - Differential increases in blood flow velocity in the middle cerebral artery after tourniquet deflation during sevoflurane, isoflurane or propofol anaesthesia. AB - The purpose of this study was to examine the comparative effects of sevoflurane, isoflurane or propofol on cerebral blood flow velocity after tourniquet deflation during orthopaedic surgery. Thirty patients undergoing elective orthopaedic surgery were randomly divided into sevoflurane, isoflurane and propofol groups. Anaesthesia was maintained with sevoflurane, isoflurane or propofol infusion in 33% oxygen and 67% nitrous oxide, in whatever concentrations were necessary to keep bispectral index values between 45 and 50. Ventilatory rate or tidal volume was adjusted to target PaCO2 of 35 mmHg. A 2.0 MHz transcranial Doppler probe was attached to the patient's head at the temporal window and mean blood flow velocity in the middle cerebral artery was continuously measured. The extremity was exsanguinated with an Esmarch bandage and the pneumatic tourniquet was inflated to a pressure of 450 mmHg. Arterial blood pressure, heart rate, velocity in the middle cerebral artery and arterial blood gas analysis were measured every minute for 10 minutes after release of the tourniquet in all three groups. Velocity in the middle cerebral artery in the three groups increased for five minutes after tourniquet deflation. Because of the different cerebrovascular effects of the three agents, the degree of increase in flow velocity in the isoflurane group was greater than in the other two groups, the change in flow velocity in the propofol group being the lowest (at three minutes after deflation 40 +/- 7%, 32 +/- 6% and 28 +/- 10% in the isoflurane, sevoflurane and propofol groups respectively, P < 0.05). PMID- 19681419 TI - A retrospective analysis of deep neck infections at Royal perth hospital. AB - Serious deep neck infections may result in life-threatening airway complications. The aim of this study was to review the management of patients requiring surgical drainage with deep neck infections and to identify possible factors that may predict a greater risk of airway complications. In this study the authors reviewed the notes of patients requiring surgical drainage of deep neck infections who were admitted to Royal Perth Hospital over a seven-year period (2000 to 2007). One hundred and twenty-nine suitable patients were identified, of whom 15.5% encountered airway complications including one death due to airway obstruction. Airway complications were more common if there was no consultant anaesthetist present (odds ratio 4.01 [confidence interval 1.20 to 13.46], P = 0.02). Deep neck infections are still relatively common and are associated with significant morbidity and mortality. Patients with deep neck infections represent an anaesthetic challenge which should be managed by those with an appropriate level of experience. PMID- 19681420 TI - Bacterial contamination in solution aspirated from non-sterile packaged fentanyl ampoules: a laboratory study. AB - Iatrogenic meningitis is a rare complication of spinal anaesthesia. It is mandatory to use a technique which minimises the risk of introducing bacteria into the subarachnoid space. Currently available fentanyl ampoules require a careful drawing-up technique, as the outside of the ampoule is not sterile and there is potential to contaminate the contents in the drawing-up process. We designed a pilot laboratory study to determine the extent of bacterial contamination of fentanyl solutions drawn up from non-sterile packaged glass fentanyl ampoules using three different methods: aspirating through a 5 microm filter needle only, aspirating through a 5 microm filter needle after swabbing the neck of the ampoule with isopropyl alcohol and aspirating through an antibacterial filter in addition to the 5 microm filter needle. Ten anaesthetists used each method once, in randomised order to draw up solution from three different fentanyl ampoules. Samples underwent bacterial culture in blood agar and enrichment broth for 48 hours. No significant growth was seen in any sample. This pilot study did not identify any bacterial contamination of fentanyl solution irrespective of which of the three methods for aspiration was used. PMID- 19681421 TI - Sixteen years of severe Tiger snake (Notechis) envenoming in Perth, Western Australia. AB - We aimed to describe the characteristics, clinical course, management and outcome of patients presenting to Perth teaching hospitals after envenoming by Tiger snakes. We undertook a chart review from six Perth teaching hospitals over a 16 year period from 1990 to 2005. Data were collected by a trained investigator using a preformatted data abstraction tool. We included patients bitten in the appropriate geographical area, with defibrination coagulopathy and positive Venom Detection Kit result for Tiger snake or response to specific antivenom. Of 381 charts reviewed, 23 patients were envenomed by a Tiger snake. The mean age was 36 years, 83% were male and all were bitten on a limb. First aid was applied poorly and all patients were symptomatic on presentation. Six patients developed rhabdomyolysis, one renal failure, four clinical bleeding, three neurotoxicity, one non-fatal respiratory arrest and one fatal cardiac arrest. All patients received antivenom, 13 received adrenaline premedication, with two mild allergic reactions developing in non-premedicated patients. The average dose of antivenom was four ampoules. Mean hospital stay was 2.6 days. This is the largest series of Tiger snake envenoming reported in Australia. Only one patient of 23 (4%) died, despite all patients being significantly envenomed. With rapid antivenom treatment and modem emergency and intensive care management, most patients envenomed by Tiger snakes survive. PMID- 19681422 TI - Adenosine-induced cardiac standstill to facilitate endovascular embolisation of cerebral arteriovenous malformations in children. AB - This report describes three children, aged eight to 11 years, with high-flow cerebral arteriovenous malformations who underwent interventional neuroradiological procedures involving glue (N-butyl cyanoacrylate) embolisation under general anaesthesia. The procedure was facilitated by relative hypotension induced by esmolol infusion and intravenous adenosine boluses. To allow controlled deposition of N-butyl cyanoacrylate into the arteriovenous malformations, glue injection was synchronised with the onset of adenosine induced brief cardiac standstill. This resulted in satisfactory obliteration of the arteriovenous malformations nidus in all cases. The haemodynamic modulations, including the adenosine-induced brief cardiac standstill, was noted to not affect the BIS values in our patients. All patients had satisfactory obliteration of their arteriovenous malformations and had good neurological outcomes at one-year follow-up. PMID- 19681423 TI - The ability of anaesthetists to identify generic medications from trade names. AB - The recent proliferation of brand names for prescription medications has made the clinician's task of identifying the corresponding generic drug substances more difficult. A survey of 86 anaesthetists and anaesthetic trainees at two Melbourne hospitals was conducted to measure the extent to which this was perceived to be a clinical problem. In addition, a theoretical test was administered to examine the ability of these anaesthetists to correctly identify generic drugs and therapeutic groups when only the brand name is provided. The results indicated this is perceived to be a genuine clinical problem, with more than 80% of respondents encountering unfamiliar trade names 'often' or 'always' and the test revealing that fewer than one third of commonly prescribed brand names were identified correctly. PMID- 19681424 TI - Survey of airway skills of surgeons in Western Australia. AB - Our objective was to survey all consultant surgeons, including obstetricians/gynaecologists, in the State of Western Australia to assess their experience with, and readiness to assist anaesthetists with a difficult or failed airway. Survey questionnaires were mailed to all surgeons registered in Western Australia (n = 445). A total of 238 responses (53%) were received, mostly from general surgeons, obstetrician/gynaecologists and orthopaedic surgeons. Forty percent had provided non-surgical assistance with a difficult airway and 60% had assisted with a surgical airway. All ear nose and throat surgeons who responded to the survey had assisted with an emergency surgical airway and 47 surgeons reported having performed six or more surgical airways. However 26% of respondents had never performed a surgical airway and 37% did not feel confident in performing an urgent surgical airway Seven percent of respondents reported witnessing a failed airway that resulted in death or neurological damage. Seventy percent of respondents had undergone formal training in tracheostomy and 26% had advanced trauma life support or early management of severe trauma training. These findings indicate that surgeons in Western Australia perform surgical airways infrequently and only occasionally assist anaesthetists with difficult airway management. However, some surgeons lack confidence and training in surgical airway management. Because anaesthetists cannot always rely on their surgical colleagues to provide a surgical airway during a crisis, we recommend that anaesthetists discuss airway management with their surgical colleagues for all patients with identified difficult airways and that anaesthesia training should include surgical airway management. PMID- 19681425 TI - A minimum cost anaesthesia machine, compliant with Australian and New Zealand College of Anaesthetists Professional Document T3, for medical missions. AB - This report describes what seems to be the simplest and least expensive anaesthesia machine possible which complies with the Australian and New Zealand College of Anaesthetists technical document T3. PMID- 19681426 TI - Web-based formative assessment case studies: role in a final year medicine two week anaesthesia course. AB - Formative Assessment Case Studies (FACS) are an e-learning resource consisting of a case scenario punctuated with decision-making steps (multiple-choice questions) and feedback for wrong answers. FACS was developed to enhance clinical decision making skills. We wrote six FACS scenarios covering preoperative assessment topics and made them accessible to 149 final year medical students as part of their two-week anaesthesia module. A data management system recorded usage and performance by each student. Eighty-one percent of students attempted FACS (six cases 53%, five cases 17%, one to four cases 9%) and 61 to 70% completed all steps. On average FACS was attempted 1.5 times. Students required 44 to 95% more steps than the minimum to complete each case. There were two patterns of use: some students completed the cases within five to seven minutes (first quartile) focusing on the questions, while others spent over 22 to 35 minutes (fourth quartile) exploring the FACS and feedback. FACS usage correlated (r2 = 0.32: P < 0.01) with written case report marks. The students' evaluation of FACS was high. FACS is an e-learning resource that is interactive and facilitates higher learning. It can be applied successfully to disciplines less well represented in the medical curriculum, such as anaesthesia. FACS facilitated our teaching of preoperative assessment to a group of final year medical students. It was well received and shown to facilitate the learning of decision-making skills. The students' usage of FACS could have been enhanced by making FACS compulsory and using summative FACS for assessment. PMID- 19681427 TI - Anaphylactoid reaction to intravenous contrast in patient with systemic mastocytosis. AB - Systemic mastocytosis is a rare disorder characterised by tissue infiltration of morphologically abnormal mast cells and has been associated with severe anaphylactoid reactions during general anaesthesia. We report the case of a 43 year-old woman who developed a severe anaphylactoid reaction to iodinated contrast media. Persistently elevated serum tryptase levels led to further evaluation and the eventual diagnosis of systemic mastocytosis. This case highlights the importance of repeated measurements of serum tryptase levels following severe anaphylactoid reactions. The anaesthetist should also be aware of the propensity of these patients to develop severe anaphylactoid reactions during general anaesthesia and use treatment strategies to minimise this risk. PMID- 19681428 TI - Application of the transversus abdominis plane block in the intensive care unit. AB - We present three patients admitted to the intensive care unit following major intra-abdominal surgery who underwent ultrasound guided transversus abdominis plane (TAP) block for postoperative analgesia. In two patients, epidural analgesia was relatively contraindicated. In the third patient, the TAP block was used to provide rescue analgesia in the setting of an ineffective epidural. We found that the TAP blocks provided satisfactory analgesia and appeared to enhance recovery by reducing morphine requirements and avoiding opioid-related side effects. We put forward a case for more frequent use of this simple and safe technique in the intensive care setting. PMID- 19681429 TI - Too much of a good thing--air, that is. PMID- 19681430 TI - Ultrasound versus neurostimulation for popliteal block: another vain effort to show a non existing clinical relevant difference. PMID- 19681431 TI - Jackson spinal table failure: a possible design flaw. PMID- 19681432 TI - A case of needle-free delivery system incompatibility. PMID- 19681433 TI - Use of light source and introducer of proseal laryngeal mask airway for direct laryngoscopy and intubation in a child with restricted mouth-opening. PMID- 19681434 TI - Laryngeal web associated with Simpson-Golabi-Behmel syndrome in a child. PMID- 19681435 TI - Iliac crest bone graft--could this be an ideal indication for a transversus abdominus plane block? PMID- 19681436 TI - Inventor of the Macintosh laryngoscope. PMID- 19681437 TI - Dose of magnesium sulphate for severe acute asthma. PMID- 19681438 TI - Deep brain stimulation for epilepsy: knowledge gained from experimental animal models. AB - Since the development of Deep Brain Stimulation (DBS) for Parkinson's Disease, DBS has been suggested as a treatment option for various other neurological disorders. Stimulation of deep brain structures for refractory epilepsy appears to be a safe treatment option with promising results. As research on the evaluation and optimization of DBS for refractory epilepsy may be difficult and unethical in patients, studies on animal models of epilepsy are indispensable. Various brain structures and specific nuclei such as the basal ganglia, the cerebellum, the locus coeruleus and temporal lobe structures have been investigated as target areas for DBS. Additionally, a wide variety of stimulation parameters are available, with a range of stimulation frequencies, pulse widths and stimulation intensities. This review provides an overview of the relevant literature on experimental animal studies of DBS for epilepsy. Knowledge gained from animal studies can be used to answer questions regarding the optimal brain targets and stimulation parameters in human applications. PMID- 19681439 TI - Action research: what, why and how? AB - Action research is a form of research that enables practitioners to investigate and evaluate their own work. It is increasingly used in health care research; it is a research strategy in which the researcher and practitioners from the setting under study work together in projects aimed at generating new knowledge and simultaneously improving practice. This article gives an overview of the theoretical background of action research, its international historical development and explanations of its varied forms and related practical applications. Ethical problems are discussed as are questions of rigour The article shows that action research can be used to bridge the gap between theory and practice by generating knowledge fitting the particular circumstances in the practical setting, thereby avoiding problems of implementation of research findings due to lack of fit or lack of motivation. Action research lastingly increases the capacities of practitioners to solve problems encountered in practice. PMID- 19681440 TI - Diagnostic and therapeutic pitfalls in neurosarcoidosis. AB - Neurosarcoidosis is a diagnostic challenge, especially in the absence of systemic involvement, even when cerebral biopsies show noncaseating granulomas. We report a patient with a pineal germinoma associated with a extensive peri- and intra- tumoural granulomatous reaction, who was first diagnosed as possible neurosarcoidosis. A second patient was initially considered as suffering from Multiple Sclerosis. Brain biopsy showed typical granulomas and gallium scintigraphy revealed other locations of the disease. Unfortunately, he developed a severe, steroid-induced, epidural lipomatosis at the Th3-Th8 levels and died unexpectedly after surgical decompression. Granulomatous inflammation in a tissue obtained by biopsy from a midline lesion should be always considered for the differential diagnosis of germinoma. Corticosteroid-sparing immunosuppressant drugs should be used early in neurosarcoidosis. PMID- 19681441 TI - Systemic and non-systemic vasculitis affecting the peripheral nerves. AB - Vasculitis affecting the peripheral nerves predominantly manifests as subacute, progressive, asymmetric sensorimotor polyneuropathy or mononeuritis multiplex, and more rarely as painful mononeuropathy, pure sensory neuropathy, neuropathy of the cranial nerves, plexopathy, or as autonomic neuropathy. Vasculitic neuropathy may occur isolated or non-isolated (systemic) together with involvement of other organs. Systemic vasculitis with involvement of the peripheral nerves is further subdivided into primary (Takayasu syndrome, giant cell arteritis, classical panarteritis nodosa, thrombangitis obliterans, Kawasaki disease, Churg-Strauss syndrome, Wegener granulomatosis, cryoglobulinemic vasculitis, Behcet disease, microscopic polyangitis, Schoenlein Henoch purpura) or secondary systemic vasculitis (autoimmune connective tissue diseases, vasculitis from infection, sarcoidosis, malignancy, drugs, radiation, or diabetes). In addition to routine laboratory investigations and nerve conduction studies, nerve biopsy is essential for diagnosing the condition and to delineate it from differentials, although its sensitivity is only approximately 60%. Therapy of non-viral vasculitic neuropathy is based on corticosteroids and cyclophosphamide alone or in combination. Additional options include azathioprine, methotrexate, mycophenolate mofetil, or rituximab. In single cases immunoglobulins, immunoadsorbtion, or plasma exchange have been successfully applied. In case of virus-associated vasculitis interferon alpha plus lamivudine or ribaverin may be beneficial. PMID- 19681443 TI - Clinical and electrophysiological evaluation of patients with thalidomide-induced neuropathy. AB - BACKGROUND: Whether thalidomide induces a sensory ganglionopathy or a length dependent axonal neuropathy is disputed. Moreover no agreement exists concerning the effects of thalidomide dosage on the clinical and electrophysiological findings. OBJECTIVE: We examined the effect of age, gender disease duration, total cumulative dose on the clinical and electrophysiologic parameters. METHODS: Fifteen patients who had previously received 100 mg/day of thalidomide for the treatment of multiple myeloma were evaluated retrospectively. Clinical findings and nerve conductions studies were evaluated using a modified total neuropathy scoring system. RESULTS: Sensory symptoms (p = 0.033, r = 0.552) and objective sensory findings (p = 0.002, r = 0.730) worsened with higher thalidomide doses. There was no effect of age, gender and disease duration, neither on clinical symptoms and objective findings, nor on electrophysiologic data. Twelve patients (80%) developed the electrophysiological findings of neuropathy. Six (40%) had pure sensory and 4 (26.6%) had sensori-motor peripheral neuropathy, while 4 (26.6%) had carpal tunnel syndrome. Sural sensory nerve action potential (SNAP) amplitudes were more prominently reduced compared to SNAPs obtained from the upper extremities. Sural SNAP amplitude showed a tendency toward reduction as the total cumulative dose, although it is not statistically significant (respectively; p = 0.187). Significantly reduced ulnar peroneal and tibial compound muscle action potential amplitudes, slow motor nerve conduction velocities of the ulnar and peroneal nerves were found in the study group compared to reference norms (p < 0.05). CONCLUSION: Our results suggest that thalidomide produces a dose dependent peripheral neuropathy, mainly localized to the peripheral nerves in a length dependent manner. The patient must be monitored closely to prevent irreversible consequences. PMID- 19681442 TI - The accuracy of hospital discharge coding for hemorrhagic stroke. AB - BACKGROUND: Hospital discharge data is used in monitoring stroke epidemiology, and ensuring adequate resource allocation to treatment programs. Previous studies have reported variable accuracy levels for such data. We present the first study assessing the accuracy of International Classification of Diseases 10th Edition (ICD-10) discharge coding for hemorrhagic stroke in England. METHODS: We identified all patients with a primary diagnosis of intracerebral hemorrhage (ICH; ICD-10 code: I61.x) and subarachnoid hemorrhage (SAH; I60.x) admitted to the Newcastle upon Tyne Hospitals from 2002-2007. Positive predictive values (PPV) were calculated through validation with patient notes. RESULTS: Hospital discharge coding identified 978 ICH and 1169 SAH admissions over the six years. The number of diagnoses verified by patient notes was excellent for both ICH (n = 938) and SAH (n = 1123), with a PPV of 95.9% for ICH (95% confidence interval, CI = 94.5-97.0%) and 96.1%for SAH (95% CI = 94.8-97.0%). The coding errors observed were largely expected, with different types of stroke miscoded as ICH and SAH. CONCLUSIONS: The accuracy of ICD-10 hospital discharge coding for hemorrhagic stroke was excellent. However further research is needed to find ways to further improve its accuracy. PMID- 19681444 TI - Tuberculous spondylitis--analysis of 22 cases. AB - Tuberculous spondylitis (TS) frequently poses both diagnostic and therapeutic challenges. The clinical symptoms, radiological imaging studies and laboratory tests are quite often inconclusive in the early stages of the disease. GOAL: To identify early clinical symptoms, review results from radiological imaging studies and laboratory tests to establish their diagnostic value and determine the effect of conservative therapy in patients with early TS. RESULTS: Twenty two (22) patients with TS subjected to conservative therapy were studied. Medullary compression syndrome was found in 10 patients. The highest diagnostic value was established by Magnetic Resonance Imaging (MRI) data for discitis engaging two adjacent vertebrae and QuantiFERON TB Gold and T SPOT.TB tests. The effect of the disease on 12 patients, whose treatment had started before the collapse of the bodies of the vertebrae was non-occurrence of any residual deformity of the spinal column. The effect on 9 patients, whose therapy started at a later stage was a recovery of the neurological deficiency and deformity occurrence of various degrees. CONCLUSION: Repetitive lateral plain radiographs, MRI and QuantiFERON TB Gold test have proven of highest diagnostic value at this stage of the disease in our patients. The presence of clinical data, accelerated ESR levels, plain radiography and MRI evidence of an inflammatory process preceding the occurrence of bone destruction and the formation of paravertebral soft tissue collection provide sufficient reasons to conduct QuantiFERON TB Gold or T SPOT.TB testing and start an antituberculous therapy. In the frequent cases when the diagnosis cannot be sufficiently proven, we recommend empirical anti-tuberculous therapy. PMID- 19681445 TI - Dystrophin Dp71f associates with components of the beta1-integrin adhesion complex in PC12 cell neurites. AB - Dystrophin Dp71 has been implicated with cognitive impairment shown by Duchenne muscular dystrophy patients. To study Dp71 neural role, we used PC12 cell line, since these cells differentiate into sympathetic like neurons when stimulated with nerve growth factor Previously in undiferentiated PC12 cells, it was demonstrated that dystrophin Dp71f is a key component of the beta1-integrin adhesion complex that confers proper complex assembly. Since integrin based mediated adhesion is important during neuronal differentiation, it was important to know if dystrophin Dp71f was a structural component of the beta1-integrin adhesion complex in neurites of nerve growth factor stimulated PC12 cells. In the present work, by performing immunofluorescence assays, we determined the association of dystrophin Dp71f with some components of the beta1-integrin adhesion complex such as beta1-integrin subunit, talin, alpha-actinin and vinculin in neurites of nerve growth factor stimulated PC12 cells seeded onto the extracellular matrix protein laminin. The association was stronger in neural growth cones suggesting that dystrophin Dp71f is important for the function that the beta1-integrin complex has during neurite outgrowth. PMID- 19681446 TI - Psychogenic non-epileptic seizures, anticipatory grief and limited access to medical care. AB - Psychogenic non-epileptic seizures (PNES) in the context of meningoencephalitis can occur with sufficient stressors. Video-EEG is a critical diagnostic tool in such complex cases. Once the diagnosis of PNES is confirmed by video-EEG, psychiatric consultation with psychotherapeutic intervention to address underlying psychopathology and specific stressors that led to these conversion symptoms is required. Clinicians need to understand the importance of cultural themes, including the economics of healthcare, and anticipatory grief as causative stressors in pseudoseizures. PMID- 19681447 TI - Hypertensive encephalopathy with atypical MRI leukoencephalopathy affecting brain stem and cerebellum. AB - Reversible posterior leukoencephalopathy syndrome associated with hypertension rarely presents with predominant involvement of the brainstem and sparing of the supratentorial regions. In this study, the clinical and neuroimaging features of a 39-year-old woman with hypertensive encephalopathy and magnetic resonance imaging (MRI) findings localized to pons and bilateral middle cerebellar peduncles were described. Reversible posterior leukoencephalopathy syndrome associated with hypertension rarely shows isolated brainstem and cerebellum involvement, and it is important to be familiar with the lack of correlation between the severity of the radiological abnormality and the clinical status. PMID- 19681448 TI - Unusual presentation of a spontaneous spinal epidural haematoma. AB - Spontaneous spinal epidural haematoma (SSEH) is a rare clinical entity that generally requires an urgent surgical evacuation. The combination of Brown Sequard syndrome (BSS) and Horner's syndrome (HS) as the presenting symptoms of a traumatic spinal epidural haematoma is very unusual, but it has never been observed in cases of spontaneous haematoma. We herein describe a case of SSEH presenting with simultaneous BSS and HS. The possibility of a conservative management in similar cases is discussed. PMID- 19681449 TI - Chronic subdural haematoma after endoscopic treatment of a supracellar arachnoid cyst. AB - Neuroendoscopy is considered a safe treatment option for intracranial arachnoid cysts. However a variety of complications has been reported after such interventions. Here we present the first case of a chronic subdural hematoma two months after the combined treatment of a supracellar arachnoid cyst with endoscopic third ventriculostomy and cyst fenestration. PMID- 19681450 TI - Isolated dissecting aneurysm of the posterior inferior cerebellar artery. PMID- 19681452 TI - Bilateral haemorrhagic infarction of the globus pallidus after cocaine and alcohol intoxication. AB - Cocaine is a risk factor for both ischemic and haemorrhagic stroke. We present the case of a 31-year-old man with bilateral ischemia of the globus pallidus after excessive alcohol and intranasal cocaine use. Drug-related globus pallidus infarctions are most often associated with heroin. Bilateral basal ganglia infarcts after the use of cocaine, without concurrent heroin use, have never been reported. In our patient, transient cardiac arrhythmia or respiratory dysfunction related to cocaine and/or ethanol use were the most likely causes of cerebral hypoperfusion. PMID- 19681451 TI - An extreme lateral lumbar disc herniation mimicking L4 schwannoma. AB - OBJECTIVE: We herein describe a case with an extraspinal mass distorting the right L4 dorsal root ganglion. Initially presumed to be a nerve root schwannoma, the lesion turned out to be a free disc fragment. METHODS AND RESULTS: A 46-year old woman presented with a history of mild low back and intense right leg pain. The leg pain was like electrical discharges. Right knee extension was weak together with hyperalgesia and loss of heat sensation in the right LA dermatome. The right patellar reflex was absent. Electromyography showed acute and chronic denervation of muscles innervated by the right L4 nerve root. MR scan showed a right L4-5 extraforaminal mass distorting the L4 dorsal root ganglion. The mass enhanced homogeneously after gadolinium injection and was thought to be a tumor. It was surgically removed using a midline incision and intraspinal, followed by extraspinal dissection. Under the operating microscope, the mass extended between an intact lateral longitudinal ligament and a swollen dorsal root ganglion. Histopathologic examination ruled out a tumor and showed that the mass consisted of degenerated disc material surrounded by a large peripheral zone of neovascularization. CONCLUSION: Extraforaminal periganglionic free, encapsulated disc fragments may mimic tumoral masses, from which they may not be distinguished on MRI. PMID- 19681453 TI - Leptomeningeal carcinomatosis: a rare complication in esophageal cancer. PMID- 19681454 TI - [Features and pathophysiology of acute coronary syndrome]. AB - Coronary heart disease (CHD) is the leading cause of death in developed countries, and it also has fastest growing death rates in developing countries. Patients with acute coronary syndromes (ACS) are divided into two groups - those with and those without ST segment elevation. ACS without ST segment elevation also includes unstable angina pectoris (UA) any non-ST elevation myocardial infarction (NSTEMI). It is important to note that UA is defined as ischemic chest pain at rest without a rise in serum cardiac biomarkers, while the establishment of NSTEMI diagnosis requires a rise in serum cardiac biomarkers. ACS with ST segment elevation is STEMI, and it includes both ST segment elevation and a rise in serum cardiac biomarkers. Connection of UA, NSTEMI and STEMI is based on the fact that these are closely connected conditions with similar pathogenesis and clinical presentation, but they do differ by the level of severity. The main difference lies in the fact whether or not the ischemia is serious enough to cause myocardial damage of such an extent that will cause the release of a sufficient amount of serum cardiac biomarkers so that these can be discovered and measured in serum (for example, cardiac troponin). The key role in ACS is played by atherosclerosis, atherosclerotic plaque and plaque rupture, in combination with thrombosis as an event of paramount importance--thrombosis. PMID- 19681455 TI - [Electrocardiography in the diagnostics of acute myocardial infarction with ST elevation]. AB - After a century of clinical use, electrocardiography (ECG) remains the basic method for the diagnosis of acute myocardial infarction. Its suboptimal sensitivity can be improved by additional recording leads and device adjustment with posterior wall inverse presentation for the detection of left ventricular posterior wall and right ventricle infarcts. The addition of some ST-depression criteria equivalent to those of ST elevation is discussed, with the aim of increasing the sensitivity without a substantial loss of specificity. ECG patterns concerning myocardial infarction evolution, localization and size estimates are reviewed. PIS and EMI patterns are discussed in respect to the modes of reperfusion therapy. ECG scoring systems are commented, considering risk stratification and the choice of reperfusion therapy. In conclusion, ECG is an old diagnostic tool that should be used for new tasks. PMID- 19681456 TI - [Biochemical markers in the diagnosis of acute coronary syndrome]. AB - Acute coronary syndrome is a set of symptoms interpreted as being the result of cardiac ischemia. The subtypes of acute coronary syndrome, depending on the degree of cardiac ischemia, include unstable angina and two forms of myocardial infarction. Determination of serum cardiac markers plays a key role in the diagnosis of acute myocardial infarction. Serum markers such as aspartate transaminase, lactate dehydrogenase, and creatine kinase are no longer used because they lack cardiac specificity and sensitivity. According to the NACB (National Academy of Clinical Biochemistry) recommendations, two serum cardiac markers need to be determined for routine diagnosis of acute myocardial infarction, i.e. one showing early elevation in serum (up to six hours after chest pain), and the other, late marker that is elevated six to nine hours after chest pain, has high sensitivity and specificity for detection of myocardial injury, and remains elevated for several days of the symptom onset. In current clinical practice, myoglobin, CKMB mass (improved diagnostic sensitivity in relation to CKMB activity) and cardiac troponins are commonly determined. CKMB mass is a cardiospecific marker, but can also be elevated in skeletal muscle damage. Myoglobin is not cardiospecific, but has high early sensitivity (fast and reliable exclusion of acute myocardial infarction) and the possibility of rapid assessment of the success of thrombolytic therapy. Cardiac troponins are late markers for the diagnosis of myocardial injury. They are markers with highest specificity and sensitivity for acute myocardial infarction. New markers such as ischemia modified albumin, heart fatty acid binding protein, glycogen phosphorylase isoenzyme BB, carboanhydrase 3, and new tehnologies are under investigation to advance our knowledge about heart disease. PMID- 19681457 TI - [Role of echocardiography in acute coronary syndrome]. AB - This article reviews the diagnostic and clinical role of echocardiography in acute coronary syndrome (ACS). The diagnosis of ACS is based on the medical history data, ECG and cardiac enzymes. Echocardiography is an accurate, inexpensive, rapid and noninvasive test, which has a diagnostic and prognostic value and detects complications in ACS. Echocardiography can evaluate damaged myocardium and myocardium which is not involved with acute ischemia, and therefore it is recommended for establishing diagnosis in patients with chest pain and clinical evidence of structural heart damage or suspected aortic dissection, for evaluation of chest pain in patients with suspected acute myocardial ischemia with unresponsive hemodynamic instability or when other methods are undiagnostic. By localizing and quantifying segmental wall motion abnormalities, echocardiography can identify the location and extent of the ischemia. It is also the mainstay of diagnosis and evaluation of mechanical complications of myocardial infarction that include ischemic mitral and tricuspid regurgitation, ventricular and papillary muscle rupture, LV thrombi, left ventricular aneurysms and pseudoaneurysms. Echocardiography is also used for risk stratification and prognosis after myocardial infarction by assessing systolic and diastolic left ventricular function. New echocardiographic methods such as stress echocardiography and myocardial contrast echocardiography have emerged and contribute to an accurate diagnosis of ACS and better evaluation of reperfusion therapy. In emergency room and coronary care units, echocardiography as a noninvasive and inexpensive method in comparison with other established methods has significant utility in the diagnosis and management of ACS patients. PMID- 19681458 TI - [Therapy of acute myocardial infarction in primary care and emergency medicine: experience acquired in the National Interventional Cardiology Program]. AB - Since the highly efficient treatment of acute myocardial infarction (AMI) depends on the time elapsed from the occurrence of symptoms, the speed and accurate diagnosis as well as risk stratification highly influence the outcome of treatment and life prognosis. The basis of prehospital diagnosis of AMI is history, clinical examination and pre-hospital 12-lead electrocardiography performed within 10 minutes from establishment of the first contact with a health institution. Care of patients suffering from AMI by emergency service or primary care physicians needs to include monitoring, use of emergency medicines and transportation to the institution that can put the most efficient treatment strategy in place depending on the length of difficulties, type and localization of AMI, and degree of risk in each individual patient. In Croatia, the integrated diagnostic-therapy-organizational systems of care for "real-world" patients with AMI within the National Interventional Cardiology Program have been applied since the mid-2005. This treatment strategy including prehospital notification and activation of cardiac catheterization laboratory uses primary percutaneous coronary intervention, aimed at more effective life saving and disability reduction than in case of traditional pharmacology-based strategy. Apart from coordination between out-of-hospital healthcare and secondary and tertiary institutions, an adequate support and financial resources provided by local community are required. The purpose of this paper is to present implementation of a modern strategy of prehospital management of all patients with AMI, which, in the conditions of integrated medical care, requires appropriate choice of treatment according to the risk. PMID- 19681459 TI - [Therapeutic procedures in patients with acute coronary syndrome and life threatening arrhythmias in prehospital stage]. AB - The prehospital phase of acute coronary syndrome can be divided into two parts: in the first part laypersons (or nobody), and later on professional medical staff are in contact with the patient. This part is especially important since it is well-known that the mortality rate arising from acute myocardial infarction is highest in the first hour of the disease onset, mostly due to malignant arrhythmias. Apart from calling the doctor, laypersons' help practically comes to resuscitation in case of cardiac arrest. In different regions of the world, there are different programs of interventions that have led to a decline in the mortality rate in this stage of the disease, which is nevertheless higher than in the overall later period (in Zagreb in 2003, 59% : 41%). One of the main reasons for this is late arrival in the hospital where proper help can be given (based on the Registry of Acute Coronary Syndrome of the City of Zagreb in 2003, in the first three hours 26% of women and 33% of men were admitted to the hospital, while figures change to 39% for women and 45% for men in the first six hours). Unfortunately, in Croatia there is no systematic program of prehospital care of patients before the arrival of medical staff although it has been proposed for years. The treatment of certain life-threatening arrhythmias in this phase of the disease depends on the type and level of arrhythmia, hemodynamic status and transportation time. The article provides a summary of different procedures in particular arrhythmias in comparison with the same arrhythmias in other situations and/or other etiologies. PMID- 19681460 TI - [Importance of telecommunication and transportation of patients with acute coronary syndrome]. AB - Primary percutaneous coronary intervention is the method of choice for reperfusion in the setting of acute coronary syndrome with ST elevation (STEMI). According to the guidelines, it is necessary to provide as many patients as possible with the opportunity of interventional treatment, even those living far from PCI capable hospitals. Every region in Croatia should develop a clear protocol for patients with acute coronary syndrome in whom primary PCI is indicated, while relying on already existing networks of healthcare providers. Therefore, the application of telemedical communication systems is useful, since it allows confirmation of the diagnosis by a specialist of internal medicine or cardiology and enables communication between physicians on the spot and physicians at a district hospital or PCI capable hospital. In order to save time, it should be attempted to transport the patient directly to the PCI capable hospital, skipping district hospital. The transport of the patient with the acute coronary syndrome is generally safe if safety protocols are followed: an educated attending healthcare professional, ambulance with required equipment and use of appropriate procedures of prehospital treatment. PMID- 19681461 TI - [Role of district hospital in providing care for patients with acute coronary syndrome]. AB - The role of a district hospital in providing care for patients with acute coronary syndrome is determined by epidemiologic and demographic characteristics, the quality of primary health care and the citizens' level of information. There are two organizational departments that participate in medical treatments as part of the complex of modern organized hospitals: Emergency Department and Coronary Care Unit. Emergency Department is very important for providing prompt and appropriate initial care. Coronary Care Unit provides continuous monitoring of all vital functions, invasive and noninvasive hemidynamic monitoring, 24-h medical attendance, and medical telephone connection with a tertiary center. The main task of medical treatments is the prevention of sudden death, and pain and discomfort relief with the aim of maximal shortening the time elapsed from the first symptom to definitive care. In patients with myocardial infarction and ST elevation, reperfusion therapy (fibrinolytic, primary PCI) should be started within the first two hours of the disease onset. In case of unstable angina pectoris and myocardial infarction without ST elevation, the targets are as follows: stabilization and passivization of the plaque (heparin, ASA, clopidogrel), medical treatment of underlying ischemia (beta blocker, calcium channel antagonist, nitrates) and initiation of secondary prevention (ACEI). The earliest possible assessment of possible risk of disease progression or potential death is of utmost importance to define therapeutic strategy at the very beginning of treatment. Modern medical treatment of acute coronary syndrome requires the following: 24-h accessibility of Emergency Medical Service, functional of medical telephone connection and continuous education of the entire medical staff. PMID- 19681462 TI - [The role of diagnostic work-up at university hospital emergency service and coronary care unit in the management of acute coronary syndrome patients]. AB - Acute coronary syndrome (ACS) represents a significant global socioeconomic problem. In the United States, 6-7 million patients present to emergency service annually for chest pain or symptoms of ACS, the diagnosis of ACS being confirmed in 20-25% of these patients.There are two groups of ACS patients, with chest pain as the main and common the presentation that basically has the same pathophysiologic substrate. The groups show differences in electrocardiogram (ECG) changes, or in the presence or absence of ST elevation that distinguishes acute coronary syndrome with ST elevation, or acute myocardial infarction with ST elevation from acute coronary syndrome without ST elevation (ACS-NSTE). Within these groups there are subgroups of patients with acute myocardial infarction without ST elevation (NSTEMI) and patients with unstable angina pectoris (UA). Subgroup distinction between UA and NSTEMI is based on the findings of elevated cardiac markers (troponin). Diagnostic procedures performed at emergency service and coronary unit include history and clinical status, ECG, laboratory testing for cardiac markers and other biochemical parameters, heart and lung x-ray, heart ultrasound, radionuclide methods, and assessment of the risk level (risk score). It should be noted that history data, ECG findings and testing for cardiac markers are of particular importance in setting the diagnosis of ACS. Other useful methods of risk assessment include TIMI degree of risk, which is in general use because of its simplicity, but is less predictive, and the Pursuit and Frisco degree of risk. Regardless of the method used, it is recommended to determine the degree of risk for every patient on admission and at discharge. PMID- 19681463 TI - [Anti-ischemic therapy in patients with STEMI or NSTEMI treated at county and university hospitals]. AB - The objective of prehospital care of patients with acute coronary syndrom (ACS) [acute ST segment elevation myocardial infarction (STEMI), acute non-ST segment elevation myocardial infarction (NSTEMI), and unstable angina (UA)], is prompt diagnosis of the acute myocardial infarction, patient's risk assessment, drug administration in order to reduce patient's pain and fear, and prevention or treatment of heart failure. In hospital treatment therapeutic procedures include reperfusion therapy, limitation of infarction zone, treatment of complications (heart failure, life-threatening arrhythmias), prevention of reinfarction, heart failure and eventually prevention of sudden cardiac death. Acute therapeutic procedures include revascularization, anti-ischemic and antithrombolytic treatment, possible surgical revascularization and treatment of complications (arrhythmias, heart failure). The patients with STEMI that present within 3-12 hours from the onset of chest pain should undergo primary percutaneous coronary intervention (PCI). In case of presentation within 3 hours from the occurrence of chest pain, the administration of thrombolytic therapy in this period is equally efficient as PCI. Regardless of reperfusion regimen, the anti-ischemics administered including nitrates (nitroglycerin); intravenous analgesics (morphine sulfate); O2 2-4 L/min; beta-adrenergic blockers; calcium channel blockers; angiotensin converting enzyme inhibitors (ACE-I); magnesium and glucose-insulin potassium have proved to be efficient as shown by study results and clinical experience. The mechanism of action of anti-ischemics includes reduction in myocardial oxygen consumption achieved by a decrease of heart frequency, reduction of systemic blood pressure and reduction in myocardial contractility by vasodilatation and consequent better myocardial oxygen supply. The outstanding results of major clinical studies are presented, and main guidelines for anti ischemic therapy of ACS adopted by the international professional associations are set forth. PMID- 19681464 TI - [Invasive approach to patients with acute coronary syndrome]. AB - Cardiovascular diseases are the leading cause of mortality in Croatia. In concordance with this epidemiologic situation, a new organization of emergency medicine and a network of invasive cardiac laboratories have been introduced throughout Croatia. Main goal of this structuring is to improve the care of patients with acute coronary syndrome (ACS). The aim of this paper is to open discussion on the optimal way of treatment in patients with ACS in our country today, in the era of interventional cardiology of the 21st century. The pathophysiology of ACS encompasses a complex atheroinflammatory and atherothrombotic process with dynamic and progressive mechanical obstruction of coronary arteries and subsequently oxygen supply-demand mismatch. Conversely, the best way to treat such patients is reperfusion therapy, a goal nowadays achieved by either antithrombotic medical therapy or percutaneous coronary intervention (PCI). The weight of evidence does support the use of primary PCI as a standard and supreme reperfusion therapy, especially in myocardial infarction with ST elevation. The logistic complexities such as triage, transportation, the development of capable interventional center working 24-hours, even in developed countries, may be a major problem to use such a practice in the whole community. In ACS with non ST elevation, problems are even broader and include the importance of using optimal revascularization procedure (even cardiac surgery), timing and concomitant medical therapy, with certain stratification of every individual. Finally, especially for our country, medical and economic resources should be used optimally in order to achieve an optimal system to treat patients with ACS. PMID- 19681465 TI - [Elective percutaneous coronary intervention after acute coronary syndrome]. AB - Elective percutaneous coronary intervention (PCI) after acute coronary syndrome (ACS), according to guidelines issued by the European Society of Cardiology (ESC) and American Heart Association/American College of Cardiology (AHA/ACC), is a therapeutic method that is indicated in patients with ACS with ST segment elevation in case of persistent signs of myocardial ischemia and with significant stenosis of coronary artery verified by coronary angiography, suitable for PCI according to the guidelines. It is also indicated for non-culprit significant stenosis of other coronary arteries which have been seen during primary PCI for ST segment elevation myocardial infarction (STEMI). After non ST segment myocardial infarction (NSTEMI) or after non-ST elevation ACS, elective coronary artery angiography is indicated in low risk patients if they have positive signs of ischemia on noninvasive tests. Depending on the results of coronary angiography, elective PCI is indicated according to ESC or AHA/ACC guidelines. The method success is assessed at three levels, i.e. by angiography, clinically and periprocedurally. PCI enables earlier and more efficient resolution of symptoms, better effort tolerance and lower rate of residual ischemia on noninvasive tests. PMID- 19681466 TI - [Invasive versus noninvasive approach in the treatment of nonstable angina pectoris and NSTEMI: pro and con]. AB - Acute coronary syndrome, although unique in its pathophysiologic mechanism, in its different clinical presentations is characterized by different diagnostic and therapeutic approaches. Conservative approach is complementary rather than competitive to modern invasive percutaneous procedures on coronary arteries. The guidelines are clear but have been subject to dynamic changes in the last several years and we can expect further changes and be open to them. Two issues are discussed in the article. The first is whether we need to be still more efficient in the invasive approach as an adjunct to medical therapy, and second is whether all patients with low risk should be submitted to diagnostic coronary angiography and when. PMID- 19681467 TI - [Primary prevention of cardiovascular disease]. AB - Cardiovascular diseases (CVD) are the leading cause of mortality in Croatia and in Europe. Primary prevention of CVD involves intervention before the onset of disease, and prevention of modifiable risk factors, i.e. cigarette smoking, hyperlipidemia, arterial hypertension, diabetes mellitus, inactivity, obesity. These risk factors are strongly associated and lead to impaired vascular endothelial function, chronic injury of endothelium, platelet activation and aggregation, atherosclerotic plaque formation, and in the end manifestation of CVD. The risk of any coronary event increases exponentially when two or more risk factors are present. Aside from conventional factors, it has been demonstrated that raised levels of C-reactive protein (CRP), cytokines, homocysteine and fibrinogen are also important promotors of the disease, pointing to partially inflammatory nature of coronary atherosclerosis. The effects of risk factors such as smoking, arterial hypertension and hyperlipidemia on vascular endothelium are proven to be reversible. According to Guidelines on Cardiovascular Disease Prevention in Clinical Practice of the European Society of Cardiology (2007), population is advised to follow the formula 0 3 5 140 5 3 0. It suggests that crucial measures in preserving cardiovascular health are as follows: no smoking (0), walking 3 km daily or 30 minutes of any moderate activity (3), blood pressure less than 140 mm Hg systolic (140), total blood cholesterol less than 5 mmol/L (5), LDL cholesterol less than 3 mmol/L (3), avoidance of overweight and diabetes (0). There are many studies proving the beneficial effects of statins and ACE inhibitors in improving endothelial function and endorsing primary prevention. PMID- 19681468 TI - [The role of statins in secondary prevention of STEMI and NSTEMI: when to start with statin treatment?]. AB - It has been reported that early use of statins in STEMI and NSTEMI as in acute coronary syndrome protects patients from recurrent ischemic events. Thus, it may reduce both short-term and long-term adverse outcomes such as subsequent cardiovascular mortality, myocardial infarction and revascularization, as well as stroke. It seems that this protective effect takes place as early as four months of treatment initiation, as reported in the MIRACL trial. The potential benefit slowly increases over time for up to 24 months of treatment initiation. Pleiotropic effects of statins, i.e. mechanisms such as improvement of endothelial dysfunction, antithrombotic and fibrinolytic as well as anti inflammatory effects are possible explanations for this early effect. Therefore, the initiation of treatment with statins during hospitalization as part of acute phase therapy is advised. Relatively few patients need to be treated in such a way to prevent one death, so this approach is also cost-effective. PMID- 19681469 TI - [Diabetes and acute coronary syndrome]. AB - Diabetes mellitus has a major impact on cardiac morbidity and mortality with three major aspects of the relationship: (I) coronary multiple thickening and its consequences, (2) autonomic neuropathy of the heart, and (3) cardiomyopathy. It is well established that diabetic patients are more likely than patients without diabetes to die after an acute coronary syndrome (ACS). The increased mortality is seen both in the acute phase and during long term follow up. The unfavorable prognosis of diabetic patients has mainly been attributed to more pronounced left ventricular dysfunction and a high likelihood of reinfarction. Many factors may contribute to this unfavorable outcome, such as severe diffuse coronary artery disease, disturbed autonomic tone, diabetic cardiomyopathy, and abnormal platelet activation and thrombin-generation that are pronounced in diabetes, as well as purely metabolic factors causing more oxygen consuming use of free fatty acids during acute myocardial ischemia. There are many evidence-based interventions in ACS patients that are applicable to diabetic patients with ACS, and there are no specific contraindications to evidence-based therapies in patients with diabetes; thrombolysis and the use of beta-blockers should be used alongside other evidence based therapies. The role of intensive intravenous insulin in diabetic patients with ACS remains to be clarified. PMID- 19681470 TI - [Secondary prevention and cardiac rehabilitation]. AB - Coronary heart disease is a chronic disease with multifactorial etiology. The risk depends on several, already well-known, risk factors such as hypertension, diabetes mellitus, dyslipidemia, smoking, obesity, physical inactivity, stress, etc. Multiple risk factors will increase the risk. The variety of risk factors requires a multidisciplinary approach--a team of different experts guided by a cardiologist who ensures an appropriate rehabilitation program. This should include secondary prevention as an important part of coronary heart disease patient treatment. It is based on the generally accepted medical, professional and scientific principles and long standing international experience. Papers written by experts and published on behalf of the World Health Organization, European Society of Cardiology, European Society of Hypertension, European Association for Cardiovascular Prevention and Rehabilitation, and other societies, define the basic characteristics of cardiac rehabilitation programes: aims, forms, content, indications and ways of implementing it in order to make it comprehensive, and to help regain the patients' temporarily lost ability to return to normal life and work. The cardiac patient will be fully educated to manage his exercise program and secondary prevention for life. Taking into account the situation that we have in our country, in relation to social structure, demography, legislation and types of health insurance, we would recommend that post-acute coronary syndrome patients undergo a cardiac rehabilitation program, which includes secondary prevention. After the education and rehabilitation program, aimed at individual needs, patients will continue under the guidance of his/her GP, consulting the specialist only when needed. In this way, the positive effects of cardiac rehabilitation will be long lasting. PMID- 19681471 TI - [Approach to coronary heart disease patients at a county hospital--Zabok General Hospital]. AB - The aim of the study was to determine health care measures taken in coronary patients hospitalized at Zabok General Hospital in order to identify the problems and accomplishments acquired during the period of observation (2000-2006). Nowadays, cardiovascular diseases are the leading cause of mortality and morbidity in the world and in this area. This fact was the main reason for monitoring patients treated at Department of Internal Medicine, Zabok General Hospital with the diagnosis of coronary heart disease. Data on all patients treated for coronary heart disease from 2000 to 2006 were analyzed. Coronary disease was diagnosed on the basis of history data, clinical examination, laboratory tests for biochemical coronary markers and electrocardiogram. Depending on the diagnosis, patients were treated at Intensive Care Unit, Cardiology Department, or were referred for emergency percutaneous coronary intervention (PCI). Patients were analyzed according to the diagnosis, age and sex, and results were graphically presented. Considering age factor, most of the patients were older than 70 and few were younger than 50. Considering sex, there were more male than female patients. The leading diagnosis was acute myocardial infarction with ST segment elevation (STEMI) in male patients and angina pectoris in female patients. A relatively small number of STEMI patients were referred for PCI. The main reason was the fact that patients came to the hospital too late, i.e. the time elapsed from symptoms onset was too long. Another reason was rather poor organization of the invasive cardiologist teams that could perform the operation 24 hours a day. This type of the work organization was introduced in 2003 in Zagorje County, with an increase in 2004 and at the Zagreb Ring level in 2005. The treatment and approach to coronary heart disease patients at our County Hospital is the same as at University Hospitals. It has been ensured by the application of guidelines on the approach and treatment of these patients, along with the organization of invasive approach and treatment through the Croatian Society of Cardiology. Additional education of the population at large is still necessary in order to minimize the treated for coronary heart disease as our final goal. PMID- 19681472 TI - [The region of Karlovac in the urgent PCI network]. AB - AIM: The aim of the study was to determine circumstances of referring patients with ST-elevation myocardial infarction (STEMI) for urgent percutaneous coronary intervention (PCI). PATIENTS AND METHODS: Different forms of acute coronary syndrome (ACS) and circumstances of referring STEMI patients to Karlovac PCI from October 2005 to February 2008 were analyzed. RESULTS: During the period of observation, 366 ACS patients were hospitalized. There were 37% of patients with unstable angina pectoris, 15% with myocardial infarction without ST-elevation (NSTEMI) and 48% with STEMI. Out of 176 STEMI patients, 52% had subacute infarction, 22% were referred for urgent PCI, 17% received fibrinolytic treatment, whereas in 9% of patients PCI and fibrinolysis were contradictory. Out of 30 patients with fibrinolysis, PCI was not carried out in 14 patients that arrived in hospital within 2-3 hours, 2 patients arrived after 6 hours, 3 patients were older than 75 and 11 patients refused invasive approach to treatment or it was personal estimate of the physician. CONCLUSION: Accordingly, too many patients with subacute STEMI arrived in hospital. The success of PCI project would be even better if it was used in patients with unstable angina pectoris and NSTEMI. PMID- 19681473 TI - [Experience in the treatment of patients with STEMI in the frame of urgent PCI Project in Medimurje County]. AB - In 2004, the mortality rate of cardiovascular disease in Medimurje County was 53%, with coronary heart disease accounting for 18.2% of cases. The need of organizing a County team for health was recognized. Five health priorities were acknowledged with coronary heart disease on the top of the list. In 2005, Medimurje County was among the first regions outside Zagreb that had launched the Project of urgent percutaneous coronary intervention (PCI) in ST-segment elevation myocardial infarction (STEMI) patients. Internal medicine physicians from the County Hospital received thorough education and printed materials on the issue were distributed to primary care physicians. A media campaign for the general population of the Medimurje County was also prepared. During the first 2 years of the project, the average pain-to-needle time in our patients was less than 4 hours. From the beginning of the project till July 2007, more than 100 patients with STEMI were treated with emergency PCI. There still are issues that have remained unsolved (how to reduce the pain-to-door and door-to-needle time, managing patients in cardiogenic shock, NSTEMI-patients, and presentation of patients with multivessel disease to cardiac surgeons). It is important to carry on with the intensive media campaign as well as with further education of patients, physicians and other health personnel. PMID- 19681474 TI - [Epidemiology of acute coronary syndrome in the City of Zagreb]. AB - AIM: Amongst cardiovascular diseases (CVD), acute coronary syndrome (ACS) is an important cause of cardiovascular mortality. The aim of the study was to examine ACS in Zagreb, to determine how it is treated, and to assess the extent to which new guidelines have been implemented in daily practice. During the last decades, great improvements have been made in the prevention and management of ACS. We wished to ascertain how these improvements have been implemented in our hospitals in the year 2003. METHODS: We carried out a retrospective study including patients in the City of Zagreb that had been discharged from the hospital with the diagnosis of ACS. The community-based Registry of Acute Coronary Syndrome for the City of Zagreb for the year 2003 served as the source of data. Records on 1,733 ACS patients were analyzed. In 1,349 hospitalized patients, the following parameters were investigated: length of hospital stay; time from onset of symptoms to arrival in hospital; rate of angiography and reperfusion and medical therapy after discharge. RESULTS: Study results showed that 1,105 (64%) patients were discharged from hospital, 244 (14%) died in hospital, and 384 (22%) died outside hospital. There were 1,136 (84%) patients with AMI, 638 (56%) with ST segment elevation MI, 391 (34%) with non-ST segment elevation MI and 107 (10%) of patients with possible MI and MI with undetermined ECG. Patient records revealed that 688 (51%) stayed in hospital between 8 and 14 days. The time from symptom onset to hospital arrival was less than 6 hours in 566 patients, 349 (44%) men and 217 (39%) women; 6 to 12 hours in 157 patients, 79 (10%) men and 78 (14%) women; and over 12 hours in 410 patients, 238 (30%) men and 172 (31%) women. The time of symptom onset was unknown in 216 patients, 127 (16%) men and 89 (16%) women. Angiography and reperfusion therapy were performed in 557 (41%) patients, 373 (47%) men, and 184 (33%) women; PCI/PCI + stent in 33% of men with ST or non ST segment elevation, and 25% of women with ST segment elevation and 19% of women with non-ST segment elevation. Fibrinolytic therapy was performed in 86 patients, 53 (7%) men and 30 (5%) women. Amongst ACS patients (N=966), 85% took aspirin, 70% beta-blockers, 69% angiotensin converting enzyme inhibitors and 63% statins. DISCUSSION AND CONCLUSION: Compared to other studies, non-ST segment elevation MI and unstable angina seemed to be less often diagnosed. Patients arrived in hospital rather late after symptom onset, and this was probably one of the reasons that reperfusion therapy could not be sufficiently applied. We also noted that physicians did not always follow the generally accepted guidelines to enable a better outcome by use of preventative measures and higher rate of effective therapy. There are important lessons to be learnt from this study for both patients and physicians. PMID- 19681475 TI - [The role of county health center in the management of patients with acute coronary syndrome]. AB - Health emergency service teams play an important role in the management of patients with acute coronary syndrome. They have to be educated, equipped, skilful and supported by the entire health care system. The role of county health center in the management of patients with acute coronary syndrome is illustrated in the article, based on the experience acquired at Medimurje County Health Center from Cakovec. The reformed Health Center activities including organization, coordination and linking of teams, population health monitoring at the local level, epidemiologic surveillance, education (active and passive, on both sides of college chair), joint diagnostic and other services, and quality control are discussed in detail. In contrast to a bureaucratic and formal one, a real and innovative reform should take account of necessary changes in the management and organization, not just in standards, rights and obligations. The management protocol for acute coronary syndrome patients is described: setting the main objective (acute coronary disease morbidity and mortality reduction), setting short-term and long-term specific goals, adoption of strategy based on the main objective (education, completion and particular programs pursuit, connecting, collaboration, quality assurance through clinical guidelines and protocols) and other elements, including dignity, leadership, teamwork, adoption and implementation of patient management protocols. PMID- 19681476 TI - [Antithrombotic therapy in the treatment of STEMI and NSTEMI patients]. AB - By establishing the Croatian network of primary PCI more patients can now be treated by this method every year. Nevertheless, therapeutic success depends on appropriate, i.e. complete antithrombotic therapy. On the other hand, about half of inhabitants of Croatia have not yet been covered with primary PCI network. Aspirin has an established role in antithrombotic therapy, demonstrated in numerous clinical studies. Thienopirydines, especially clopidogrel, have improved the outcome of interventional cardiology. GP IIb/IIIa receptor inhibitors, despite their high price, pave their way into our daily practice. Heparin is an essential part of therapy for almost all acute coronary syndrome scenarios, but the role of low-molecular heparins is yet to be clarified. Fibrinolytic therapy still "saves lifes". By improving new fibrinolytics, they have become easier to use, entail less side effects and are more efficient. PMID- 19681477 TI - European Federation of Oto-Rhino-Laryngology Societies (EU.F.O.S.)--past, present and future. AB - The author presents the history, the current status and the future of European Federation of Oto-Rhino-Laryngology Societes. The new European Confederation (EC ORL/HNS), a scientific and educational organization will be devoted to the advancement of science, research and education on ORL, promote, co-orrdinate and unify the advancement of ORL-HNS in Europe, organize and co-ordinate high quality training and exchange programmes and represent ORL-HNS in general. PMID- 19681478 TI - [Reconstruction after maxillectomy using the temporalis muscle flap--our experience]. AB - THE AIM OF STUDY: The study presents a method of a reconstruction of the palatum after maxillectomy. This method is choosen in selected cases of malignant tumors of the maxilla. MATERIAL AND METHODS: The method is combined of preparation of the temporalis muscle flap and its dislocation it under arcus of zygoma into the oral cavity. The surface of the flap covered with temporal fascia is oriented into the oral cavity and has been stitched in the hole of the palatum. The surface of the flap covered with periostium is connected to postoperative cavity of the maxilla. The flap covered the palatum tightly divides oral cavity from postoperative cavity of the maxilla. The vascular pedicle of the flap, with deep temporal vessels supply blood for proper healing of the palatum. Oncologic control of the postoperative cavity is performed using imaging investigation (computer tomography, magnetic resonance). The loss of tissue in the temporal fossa is covered by temporal fossa fat or synthetic material. RESULTS AND CONCLUSIONS: The reconstruction method brings satisfactory functional result. The method allows to avoid using inconvenient prostheses--obturators of palatum--in patients after maxillectomy. Complications in this method as postoperative perforation of the palatum and necrosis of the muscle flap are rare--less than 5% cases. PMID- 19681479 TI - [Endoscopic sinus balloon catherisation--evolution or revolution in treatment of rhinosinusitis? Our experience and literature review]. AB - Sinus ostium patency is a key point in treatment of rhinosinusitis. Thanks to minimally invasive set of tools dilation of ostium without tissue removal is possible that was demonstrated in large multicenter studies. The aim of our study is to present endoscopic balloon dilatation of sinuses technique under fluoroscopy and asses efficacy and safety in surgical treatment of chronic rhinosinusitis based on own experience (10 cases) and literature data. We concluded that endoscopic balloon dilation of sinus procedure is much more safe than previous techniques and increases postoperative patients comfort. PMID- 19681480 TI - [Abscess of the parotid gland]. AB - Inflammatory processes developing in the region of parotid gland can be specific or non-specific. Non-specific inflammation of bacterial or viral origin, which almost always are secondary to inflammatory processes in the region of oral cavity and pharynx, can be caused by presence of sialoliths in parotid gland or in the excretory duct. In the paper we presented 11 patients treated for the reason of parotid gland abscesses. Etiology, the course of the disease, and the method of treatment were analyzed. PMID- 19681481 TI - [Reconstruction of the nose after oncological operations]. AB - INTRODUCTION: Skin cancer (basal cell carcinoma, squamous cell carcinoma) and melanoma are the most frequent malignancy appearing in a human begin. The treatment of choice is surgical excision of tumor with margins of healthy tissue. An extent of resection determines the method of reconstruction. MATERIAL: The authors present reconstructive methods of nasal skin cancer defects performed in 61 patients treated in the Department of Otolaryngology of Medical University of Warsaw between 1998 and 2007. Basal cell carcinoma, squamous cell carcinoma and melanoma were diagnosed in 49, 10 and 2 patients respectively. Depending on the extend of tumor resection different reconstructive techniques were performed. RESULTS: In 9 patient tumor recurrence was detected during 1-2 years of follow up. Among them, in 7 patients another reconstruction procedure was required, and 1 patient underwent several surgical procedures due to subsequent recurrences. CONCLUSIONS: Surgical wounds after resection of skin cancer of the nose can be closed primarily or by means of variety of reconstructive techniques that in some cases may be combined depending on the extent of a defect. PMID- 19681482 TI - [Clinical ocular-motor disturbances in multiple sclerosis]. AB - Otoneurologic bedside examination with testing eye movements gives valuable information about static and dynamic properties of balance system and may give topodiagnostyc information about the side of lesion in patients with vertigo, dizziness and disequilibrium. THE AIM OF THE STUDY: was to present the scheme of otoneurological bedside examination and usefulness of ocular motor disturbances index in evaluation of Multiple Sclerosis patients status. MATERIAL AND METHODS: Sixty patients with diagnosis of MS, seen in outpatient neurology clinic, Medical University of Lodz, from 2002 to 2004, were enrolled into the study. Patient's history of vertigo, dizziness, hearing loss and vision disturbances were evaluated. The clinical bedside ocularmotor examination was performed in all patients. It was composed of seven tests on the basis on which we introduce ocular motor disturbances index--IRZ. RESULTS: The most frequent abnormalities were found in clinical saccadic test in 30% and smooth pursuit in 22%. MS patients who had in clinical eye movements examination IRZ bigger than 3 point formed the abnormal clinical examination group' (ACE)-- 31.7%. In u 68.3% the index was less than normal clinical examination group' (NCE). The longer duration of the disease was observed in ACE group. Comparisons of functional neurological score EDSS and for cerebellar and brainstem subscale were significantly greater in ACE group. CONCLUSION: Otoneurological bedside examinations with dynamic tests and introducing quantitative the ocular motor disturbances index is a valuable method of evaluation of visual-ocularmotor reflex and may be used in monitoring MS course of disease. PMID- 19681483 TI - [Expression of chosen chemokine receptors on the Th lymphocytes in children with hypertrophied adenoids with otitis media with effusion]. AB - INTRODUCTION: Chemokine receptors play a crucial role in the recruitment of leucocytes into inflamed tissue from secondary lymphoid organs. AIM OF THIS STUDY: was evaluation of the percentage Th (CD4+) lymphocytes with expression of chemokine receptors: CCR3, CCR4, and CCR5 in hypertrophied adenois tissue in children with otitis media with effusion. MATERIAL AND METHODS: 36 children with otitis media with effusion and 25 children with hypertrophied adenoids were tested. Expression of chemokine receptors CCR3, CCR4 and CCR5 on T CD4 lymphocytes of hypertrophied adenoid's tissue was estimated by flow cytometry method. RESULTS: Average percentage of T CD4+ lymphocytes with expression CCR4 in hypertrophied adenoid in children group with otitis media with effusion (OME = 64.11%) was significantly higher than in comparative group with hypertrophied adenoid (HA = 75.05%, p < 0.04). We showed statistically lower percentage of CD3+CD4+ cells with expression CCR3 in examinated group (OME = 47.19%) than HA group (62.66%; p < 0.003). CONCLUSION: Results suggests that intensify of humoral immunological answer in hypertrophied adenoid depends on Th2 lymphocytes with expression CCR3 and CCR4 chemokine receptors in course of inflammatory state in middle ear. PMID- 19681484 TI - [Prognostic significance of cyclin D1 and cyclin B expression in laryngeal squamous cell carcinomas treated with radiotherapy]. AB - Cyclin B1 and cyclin D1 playing the role of regulatory subunits of cyclin dependent kinases responsible for progression of cell cycle are involved in carcinogenesis of head and neck tumors. The aim of this study is to investigate their value for predicting clinical outcome after radiotherapy of laryngeal squamous cell cancer (LSCC). MATERIAL AND METHODS: The study included 50 patients with LSCC treated between the years 1998 and 2003 with radiotherapy. Tissue samples were studied immunohistochemically for detection of cyclin B1 and cyclin D1 and their expression were correlated with clinicopathological factors, local tumor control and patient survival. RESULTS: Accumulation of cyclin D1 and cyclin B1 were detected in 36% and 48% of tumors respectively. No relationship was observed between immunostaining for analyzed proteins and clinicopathologic factors. Cyclin B1 overexpression was associated with higher rate of locoregional recurrence. Five-year disease free survival rate for patients with cyclin B1 positive tumors was 73% v. 84% for cyclin B1 negative patients (p=0.005). CONCLUSION: The expression of cyclin D1 in LSCC does not seem to have a prognostic significance. Overexpression of cyclin B1 may be an indicator of the risk of locoregional recurrence in patient receiving radiotherapy. Thus cyclin B1 detected by immunohistochemistry can be useful for predicting outcome of radiotherapy in patients with LSCC. PMID- 19681485 TI - [Nutritional status of patients with cancer of larynx and hypopharyx]. AB - THE AIM OF THE STUDY: The meaning of proper nutritional state in the group of cancer patients in preoperative period was noticed already over 40 years ago. In spite, that treatment of malnutrition is easy, many patients are not diagnosed and not treated, what has obvious consequences. The aim of this study was to evaluate nutritional status of patients with cancer of larynx and hypopharynx before the treatment. MATERIAL AND METHOD: 223 men and 29 women with diagnosed squamous cell cancer of larynx and hypopharynx, who were classified to surgical treatment, were included into study. The evaluated population was divided into groups depending on location and local extend of the tumor. The nutritional state was assessed using: body mass index (BMI) and total lymphocyte count (TCL). RESULTS: Using BMI the malnutrition was diagnosed in 41% of all patients. The level of malnutrition increases while increases stage of local extend of tumor. The least numerous group of malnutritioned patients were those with tumor of glottis, but also in this group was the largest percentage of patients with local extend of tumor: T1 and T2. The state of considerable malnutrition was diagnosed in 34% patients with tumor of the piriform recess. Using TLC the malnutrition characterized 37% of all patients. In the group with tumor of glottis and epiglottis the level of malnutrition increases while increases stage of local extend of tumor. All the patients with tumor T4 with unclassified origin location were classified to the group of malnutritioned. CONCLUSIONS: The malnutrition is diagnosed in considerable percentage of patients with cancer of larynx and hypopharynx. The diagnosis of poor nutritional state will permit to apply suitable, controlled nutrition before surgical treatment. This will contribute in decrease of complication frequency and improve the tolerance of treatment. PMID- 19681486 TI - [Evaluation of apoptosis expression in granulation tissue in chronic otitis media]. AB - INTRODUCTION: There is a growing evidence that molecular and cellular mechanisms may play a role in pathogenesis in chronic otitis media. THE AIM OF THE STUDY: was to determine the intensity of apoptosis in granulation tissue in chronic otitis media. MATERIAL AND METHOD: Fifty four patients with chronic otitis media, who underwent surgical treatment, were enrolled into the study. The apoptosis was measured in paraffin-embedded granulation tissue specimens by an immunohistochemical methods, by staining with a monoclonal antibody against apo 1/Fas/CD95 and P53 protein. The bacteriological evaluation of middle ear discharges were also done. RESULTS: It was found statistically significant difference in expression of apo-1/Fas antigen between the groups with good clinical course (good healing and without recurrence) than those in the group with poor healing and recurrence (mean percentage of immunopositive cells 1.52 vs 3.34 respectively, p<0.001). The activity of apo-1/Fas antigen was more intense in tissue samples from the group with bacterial infection caused by Pseudomonas aeruginosa/Proteus sp/Staphyloccocus MRSA than those in the group without this infection (mean percentage of immunopositive cells 3.78 vs 1.75 respectively, p<0.001). The differences were also observed for P53 protein expression between the same groups, however they were not significant. There was no differences between the groups of patients with granulomatous and cholesteatomatous chronic otitis media. The significant negative correlation was found between expression of apo-1/Fas antigen and expression of P53 protein (r=-0.64, p<0.001). CONCLUSIONS: In granulation tissue in chronic otitis media different expression of apo-1/Fas antigen was found in relationship to clinical course of disease and bacterial infection caused by Pseudomonas aeruginosa/Proteus sp/Staphyloccocus MRSA. It may suggest that apoptosis mediated by apo-1/Fas mechanism may contribute to pathogenesis of chronic otitis media. PMID- 19681487 TI - [Cat scratch disease--a diagnostic problem, case report]. AB - AIM OF THE STUDY: Cat scratch disease as a possible cause of neck limphadenopathy. MATERIAL AND METHODS: We present a case of 40-years old men operated in our department on regional unilateral lymphadenopathy. The diagnosis of cat scratch disease was confirmed by the histopathologic examination. CONCLUSIONS: Regional lymphadenopathy with history of contact with cats or other animals suggests the diagnosis of cat scratch disease. If we considered this disease in differential diagnosis it would mean less traumatic treatment for the patient. Cat scratch disease is diagnosed in Poland very rarely; we need to spread the knowledge about this infectious disease. PMID- 19681488 TI - [Papillary carcinoma in thyroglossal duct cyst]. AB - Authors present a rare case of the papillary thyroid carcinoma in thyroglossal duct cyst in a 40 year-old woman. Preoperative assessment (palpation and ultrasonography) showed symptoms of typical thyroglossal duct cyst and the final diagnosis was established after histopathological examination of the surgical specimen. Epidemiology, diagnostic methods and treatment of this rare disease were also discussed in the study. PMID- 19681489 TI - [Eagle's syndrome--report of rare case of bilateral elongation of styloid proceses]. AB - Eagle's syndrome associated with elongation of styloid process or mineralization of styloid complex is uncommon pathology. The etiology of this disease has not yet be known. Main symptom of Eagle's syndrome is cervical, facial and pharyngeal pain. The authors present a case of man who suffered from severe bilateral cervical pain. After the diagnosis of both side elongated styloid process syndrome, surgical treatment was conducted. Extraoral resection of syloid processes resulted in complete and lasting pain relief. PMID- 19681490 TI - [Malignant Triton tumor of the nose--case report]. AB - Malignant Triton tumor (MTT) is classified as a variant of a malignant peripherial nerve sheath tumor with a focal rhabdomyosarcomatous differentiation. Nearly one third of described MTTs were located at the head and neck region. In following paper we described a case of MTT of the external nose in a 24 year old man. The tumor was totally exised surgically with the broad margins. Histopathological diagnosis of the tumor was made on the surgical specimen, based on immunohistochemical analysis. Radiation therapy as the next step of treatment of choice was performed in presented case. PMID- 19681491 TI - [Vestibular function in cochlear implant candidates]. AB - INTRODUCTION: The main problem of a deaf patient is per se communication process. Some of the patients, who are candidates to cochlear implantation report vertigo or imbalance in some everyday situations. The relation of patients' complaints to vestibular loss was evaluated. The authors used electronystagmography which was realized in diagnostic process to cochlear implantation. The results come from the patients diagnosed in our department in 2006-2007. In accordance to the literature, the true vertigo, was rather rare. MATERIAL AND METHODS: Medical history, caloric tests as electronystagmography results were evaluated retrospectively in 47 patients qualified for cochlear implantation in 2006-2007. RESULTS: In the study group, 25 patients did not complain of vertigo; with symmetrical calorics in 10 subjects (40%). The true vertigo reported 6 subjects; 8 subjects had imbalance and unsteadiness on walking. In the group of 47 subjects 8 of them complainted the both- vertigo and imbalance. In the ENG of this group the results of symmetrical responces were evaluated in 57% cases and vestibular hypofunction in 43% subjects. In the group with vertigo (6), 4 subjects (66%) had diagnosed unilateral hypofunction, 1 (17%) bilateral areflexion, 1 subject (17%) had symmetrical responses. CONCLUSIONS: The necessity of the confrontation ENG results to additional examination in audiological part of diagnostic process to cochlear implantation is underlined. PMID- 19681493 TI - [Vibrations of the human tympanic membrane measured with Laser Doppler Vibrometer]. AB - The knowledge of the physiology of the normal ear is important to understand the function of the ear. It is especially crucial in the reconstruction of the destroyed ear to apply the knowledge of the normal ear. We present results of tympanic membrane vibrations measurements using Laser Doppler Vibrometer in human temporal bone specimens. Six temporal bone specimens were harvested within 48 hours of death and stored cooled until preparation. The preparation included mastoidectomy with posterior tympanotomy and partial resection of the facial nerve to visualize the stapes with its footplate. We measured velocity and displacement of each quadrant of the tympanic membrane and the umbo with the laser Vibrometer equipped with velocity and displacement decoders. The sensor head OFV-534 produced and read the reflected laser beam directed at a measured point with a dedicated micromanipulator attached to an operating microscope. A retro-reflective tape was used to enhance the reflection of the laser beam. Vibrations were induced by a acoustic stimulation at the tympanic membrane. The results of the measurements were corrected to a sound pressure in the external ear canal. Laser Doppler Vibrometer system allows an undisturbed measurement of vibrations in the middle ear. Posterior quadrants of the tympanic membrane have greater velocity and displacement than anterior quadrants in lower frequencies up to 2 kHz. PMID- 19681492 TI - [Distortion product otoacoustic emission levels and input/output-growth functions in normal-hearing individuals with tinnitus and/or hyperacusis]. AB - The purpose of this study was to determine whether distortion product otoacoustic emissions (DPOAEs) can be used to distinguish among four groups with audiometrically normal-hearing sensitivity: (1) control adults without tinnitus or hyperacusis, (2) patients with tinnitus alone, (3) patients with hyperacusis alone, and (4) patients with both tinnitus and hyperacusis. Two types of DPOAE measures were evaluated: (1) the distortion product (DP-)gram measured with fixed primary levels as a function of frequency, and (2) DPOAE input/output (I/O) functions for a range of primary levels between 45 and 70 dB sound pressure level (SPL) at 1000, 2000, and 4000 Hz. DP-grams did not clearly distinguish between the control and patient groups. There was, however, a consistent trend for the three patient groups to have decreased average DP levels at 4000 and 6000 Hz; this notch in the DP-gram was not observed in the response configuration for the control group. In the three patient groups, 51 to 74% of these individuals had DP levels that were outside of the 95% confidence range for the control group. The average slopes of the I/O growth functions for each of the patient groups were consistently steeper than those for the control group; however, the slope values were indistinguishable among the patient groups. About 60% of the patients' DPOAE responses (in each group) were categorized as abnormal based on their slope values. Thus, DPOAE measures can be used with at least partial success to distinguish controls from patients with tinnitus, hyperacusis, or both tinnitus and hyperacusis, but not to discriminate among the respective patient groups. These findings suggest that the pathology represented among the patient groups is consistent at the level of the cochlea; however, diagnostic tests targeted at higher centers of processing are needed if the individuals in these groups are to be distinguished among themselves. In all participants, DPOAEs with fixed primary levels as a function of frequency were measured. There was a consistent trend for the three patient groups to have decreased average DP-gram levels at 4000 and 6000 Hz; this notch in the DP-gram was not observed in the response configuration for the control group. In the three patient groups, 51 to 74% of these individuals had DP levels that were outside of the 95% confidence range for the control group. Thus, DPOAE measures can be used with at least partial success to distinguish controls from patients with tinnitus, hyperacusis, or both tinnitus and hyperacusis, but not to discriminate among the respective patient groups. These findings suggest that the pathology represented among the patient groups is consistent at the level of the cochlea; however, diagnostic tests targeted at higher centers of processing are needed if the individuals in these groups are to be distinguished among themselves. PMID- 19681494 TI - [Deafness and mentality in Francisco Goya's paintings]. AB - The famous painter Francisco Goya y Lucientes (1746-1828) suffered during his life one or several diseases, the nature of which has not been determined with certainty. At age of 46, Goya suffered from severe illness that lasted a few months. It caused loss of vision and hearing, tinnitus, dizziness, a right-sides paralysis, weakness and general malaise. Although he recovered from a cerebral stroke which accompanied it, the deafness remained unaltered. The illness divides Goya's artistic life into two great different periods. After in the painter produces his greatest works. The visual experience after the illness was heightened by the exclusion of acoustics stimuli and the artist's talent rose to the highest level. His character became more withdrawn and introspective and his entire vitality was direscted to his painting. Goya's painting became progressively more gloom and satirical during his long convalescence. The artist suffered a stroke at age 73 that again rendered him paralysed on the right side. The precise cause of his illness has long been debated. Ome medical writers have favored the diagnosis of syphilis, some consider the possibility of an exogenous psychosis, and other suggests that the symptoms of the illness are more congruent with heavy metal poisoning, particulary lead. It is interesting to speculate how the Goya's deafness influenced the artist's mentality and the changes of his painting. Interesting is also the problem of the sensory compensation in fact of disturbance of physiological function of one of the senses. PMID- 19681495 TI - [The Jozef Czekierski's (1777-1827) otorhinolaryngological connexions]. AB - The professional and scientific activities of Jozef Czekierski (1777-1827), an eminent Varsovian surgeon on the turn of the 18th century are described widely. He was the head of surgical and obstetricial departments in Warsaw. He was also the head of the first Surgical Clinic in Warsaw. In the four-part textbook "Surgery" published in 1817-1818 Czekierski presented surgical treatment of such otorhinolaryngoogical diseases as wounds of ear and tongue, traumas of nase, diseases of parotis, the foreign bodies at respiratory tract and at the upper part of alimentary canal with full particulars. He described such rhinolaryngological operations as plastic reconstruction of the nase, tracheotomy, conicotomy, oesophagotomy in some more detail. PMID- 19681496 TI - [Joseph Toynbee--otologist, scientist, philanthropist]. AB - Joseph Tonbee's life's work may be summarized in the words of William Wilde: "The labours and investigations of Mr. Toynbee have affected more for aural pathology than those of all his predecessors either in England or on the continent". Some idea of the extent of his researches is given by the fact that he dissected some 2.000 ears. These preparations formed the Toynbee Collection in the Museum of the Royal College of Surgeons of England. He wrote the results of his researches in a catalogue that includes the description of 1,659 human ears. This catalogue forms the basis of modern otology for all time. This purely pathological catalogue was completed in 1860 with the publication of a more clinical book, "The Diseases of the Ear: their Nature, Diagnosis and Treatment". Toynbee appears to have been the first to describe the pathological changes in otosclerosis. In his book "Diseases of the Ear" he described the condition clearly, recognizing "anchylosis of the stapes to the fenestra ovalis" in 136 temporal bones. He also aligned the subjective, visual and ausculatory tests for Eustacchian tubal patency which we use today. Adam Politzer wrote: "Toynbee was the first who realized in otology that therapeutic progress depends on the knowledge of anatomy". But Toynbee was also active on other fields. Politzer, in his lecture in Vienna in 1914 said: "Toynbee was as outstanding a savant as he was a philanthropist. In addition to his scientific activity, he considered it a sacred task to dedicate his spare time to the improvement of living and health conditions of the poorer classes." Tragic enough, Toynbee's zeal for clinical experimentation went too far. Seeking to help his patients by devising a treatment to allay their tinnitus, he conceived the idea of introducing a mixture of chloroform and prussic acid into the tympanic cavity by means of Valsalva maneuver. When he made the first trial on July 7, 1866, with himself as a subject, he was found dead on the couch in his consulting room. PMID- 19681497 TI - [Hearing improvement in patients operated on chronic otitis media]. PMID- 19681498 TI - [The influence of the systemic glucocorticosteroid therapy and intranasal surgery on the taste and smell senses in patients with nasal polyps]. PMID- 19681499 TI - [Professor Waclaw Kusnierczyk (1908-1997)--Pro Memoria in the century of birthday]. AB - Waclaw Kusnierczyk was born in 1908 in Sniatyn. He received the degree in medicine at Jan Kazimierz University in Lwow in 1932. He did his PhD degree under Professor Zaleski supervision in 1938 at Jan Kazimierz University. At that time he concentrated his scientific activity on research on tuberculosis. In 1953 he obtained the title of second degree specialist in ear, nose and throat diseases. He became a chief of Otolaryngology at Urban Hospital No 4 in Katowice in 1960. Since then this eminent physician was working on tumours located in upper respiratory tract and the possibility of its endoscopic diagnosis at Silesian Academy of Medicine in Katowice. As one of the first he pointed out the negative influence of smoking cigarettes on cancer of larynx. It was Waclaw Kusnierczyk who implemented new priorities for integrated programs in patient care, research, education and cancer prevention. He has published widely in peer reviewed journals and has edited or contributed to many books. He has given many major lectures and is the recipient of numerous prestigious awards for his scientific accomplishments. The achievement of Professor Kusnierczyk were the valuable source of information for the physicians. In 1997, on the 31st of January he died in Katowice. PMID- 19681511 TI - Comparison of LASIK with the OPDCAT or OATz algorithm using the NIDEK EC-5000CXII excimer laser. AB - PURPOSE: To compare refractive outcomes, higher order aberrations, visual quality, and patient satisfaction between aspheric and whole-eye wavefront aspheric LASIK algorithms. METHODS: Two hundred seventy-four eyes of 152 patients undergoing LASIK for myopia and myopic astigmatism were divided into two groups: eyes that underwent treatment using either the OPD-guided customized aspheric treatment (OPDCAT) or optimized aspheric treatment zone (OATz). Both groups were subdivided into two groups based on preoperative manifest refraction spherical equivalent (MRSE) or = - 6.00 D. Both groups were additionally subdivided into eyes with preoperative ocular higher order aberrations <0.40 microm and eyes with higher order aberrations >or = 0.40 microm. A P value <.05 was considered statistically significant. RESULTS: At 3 months postoperatively, 88.3% (242/274) of eyes were available for follow-up. Postoperatively, 91.4% of eyes in the OPDCAT group and 90.6% of eyes in the OATz group were within 0.50 D. No difference in refractive outcomes and patient satisfaction among groups or subgroups was noted (P>.05). A significantly less change in asphericity (less oblate) was noted for the OPDCAT group (0.31 +/- 0.30) compared with the OATz group (0.51 +/- 0.35) (P<.05). A lower induction of aberrations in the OPDCAT group compared with the OATz group was noted ( P<.05). Mesopic contrast sensitivity was significantly higher for the OPDCAT groups and subgroups (P<.05). CONCLUSIONS: Refractive outcomes between groups or subgroups were equivalent. A significantly lower induction of higher order aberrations and less change in asphericity in the OPDCAT group was noted. The OPDCAT algorithm was more likely to maintain mesopic contrast sensitivity. PMID- 19681512 TI - Out of the mainstream. PMID- 19681516 TI - Mandated exercise versus free will. PMID- 19681517 TI - Clinical implications of chirality and stereochemistry in psychopharmacology. AB - Chirality, the concept of nonsuperimposable mirror images, is a fundamental property of biological systems and can be observed on a molecular, cellular, or organism level. Stereoisomer compounds possess the same molecular and structural formula, but they differ in their three-dimensional configurations. Chiral compounds have two mirror-image stereoisomer forms called enantiomers. Compounds containing mirror-image enantiomers in equal proportions are referred to as racemic mixtures or racemates. Racemates and their individual enantiomers can have very different pharmacological properties that are relevant in clinical psychopharmacology. Various examples of drug therapies that show the clinical importance of chirality and stereochemistry are described. PMID- 19681518 TI - Understanding and promoting attachment. AB - Interest in early relationships has led to increased use of terms such as attachment disorder, attachment problems, and attachment therapy when describing behavioral/emotional regulation in young children. Unfortunately, such terms are vague and lead to clinical confusion and diagnostic inaccuracies. This article will introduce attachment theory, with a discussion of reactive attachment disorder and implications for treatment of children who have problems with social emotional development. PMID- 19681519 TI - Common ground, not a battle ground. Violence prevention at a detoxification facility. AB - This article evaluates the results of a workplace violence prevention program implemented in a Colorado detoxification facility. The program interventions are modeled after federal Occupational Safety and Health Administration guidelines and use theories from both nursing and criminology for philosophy and direction. Serving as its own control, the detoxification facility shares data measured over a 4-year period, demonstrating a sharp decline in assault rates after program implementation. The importance of administrative controls, environmental adjustments, recordkeeping and evaluation, and education and training are emphasized as key components of success. PMID- 19681520 TI - Disorder or delusion? Living with Morgellons disease. AB - Whether Morgellons disease is a delusional disorder or even a disease has been a mystery for more than 300 years. Symptoms of Morgellons include crawling and stinging sensations, feeling of "bugs" and/or fiber-like material beneath the skin, disabling fatigue, and memory loss. The cause, transmission, and treatment are unknown. Research about Morgellons is staggeringly sparse and limited in scope. However, in recent years, discussion about Morgellons has become more common because of the Internet and online support groups. Mental health professionals and the general public need to be aware of the signs, symptoms, and treatment of this disease. Focusing on the disease and listening to patients can make a difference in the way health care professionals provide the best possible care for people with Morgellons. PMID- 19681521 TI - Treatment of conversion disorder. A clinical and holistic approach. AB - Conversion disorder is rare, but when it affects a young woman on the verge of adulthood, it can be devastating. The intent of this article is to encourage others by describing the success that psychiatry and alternative medicine can offer to patients with conversion disorder and to emphasize the value of nursing as a part of that team. This article will explore the attitude change in both staff and patient that was needed to achieve the ultimate goal of wellness. PMID- 19681529 TI - A commentary on information technology competencies. PMID- 19681528 TI - Clinical education reform: reenvisioning the workforce. PMID- 19681531 TI - Cultivating authentic concern: exploring how Norwegian students learn this key nursing skill. AB - The importance of interpersonal and expressive qualities has long been recognized in nursing. Drawing on data from a qualitative study among nursing students in Norway, this article discusses how nursing students cultivate authentic concern with patients. Because the nursing role has become more ambiguous, it must be created and formed in a personal way. Nursing students need to learn how to use their subjectivity in a way that promotes caring and compassionate conduct. PMID- 19681530 TI - Clinical instruction: using the strengths-based approach with nursing students. AB - Clinical instruction experience can vary significantly based on the needs of the organization and the individual characteristics of instructors and students. Clinical instructors may encounter difficulties in their relationships with students, such as personality conflicts, differences in style and values, and limited skill levels or a lack of interest on the part of students. To reduce obstacles when working with challenging students, a strengths perspective approach is recommended. This framework emphasizes discovering, affirming, and enhancing the capabilities, interests, knowledge, resources, goals, and objectives of individuals. The strengths perspective can provide an innovative framework for working with nursing students, one that emphasizes student empowerment, collaborative learning, and mutual growth. Strength-based strategies for supervision of students in clinical placements are shared, highlighting the practical application of the framework's tenets. PMID- 19681532 TI - Practicum and clinical experiences: postpracticum students' views. AB - The clinical or practicum component of preservice undergraduate education across all disciplines typically is rated by prebaccalaureate students as the most important phase of their entire professional preparation. This study collected e mail survey responses from 63 post-practicum nursing students who had just completed their culminating fourth-year clinical course. Students identified the most positive and the most negative aspects of that final practicum experience. These responses were compared with those reported by postpracticum students from the disciplines of engineering and teacher education. The cross-disciplinary similarity of these data related to postpracticum students' perceptions of this experiential learning phase of their preservice professional training was noted. The student voice provides a critical dimension to the program-enhancement process for all professional disciplines, and practicum organizers across the disciplines should value such student input and collaborate to improve the clinical phase of preservice education for all professions. PMID- 19681533 TI - The influence of concept-based learning activities on students' clinical judgment development. AB - The traditional nursing clinical education model of total patient care is becoming inadequate. New models are needed to foster deeper clinical thinking, thereby affecting students' development of clinical judgment. Concept-based learning activities, first introduced in 1990, offer a focus on a specific concept. This study evaluated the effect of concept-based learning activities on the development of clinical judgment in baccalaureate nursing students. The clinical judgment of students who were and were not exposed to concept-based learning activities was compared. Quantitative data were analyzed using a univariate analysis. In addition, a focus group consisting of members of the treatment group provided qualitative data. Results suggest concept-based learning activities are a clinical learning strategy that should be considered by faculty to deepen clinical thinking in preparation for reaching sound clinical judgments. PMID- 19681535 TI - An integrated mental health clinical rotation. AB - The most common site for accessing mental health care is the primary care setting. Yet, primary care nurses are not adequately prepared to treat the complex mental health needs of these patients. Similarly, providers in segregated mental health sites do not adequately address physical health needs. New educational models are needed to better prepare nursing graduates to provide holistic care. The integrated mental health model, which colocates mental health specialists in primary care sites, is designed to do this. This article describes key curricular elements of a successful interprofessional clinical rotation within an integrated mental health team that included the use of case studies, a standardized mental health screening instrument, a quality improvement process, and a patient satisfaction questionnaire. Family nurse practitioner and psychiatric mental health nurse practitioner students learned to collaborate with each other and with other members of the interprofessional team to provide holistic care. PMID- 19681534 TI - Five recommendations for prelicensure clinical nursing education. AB - The acute shortage of RNs is both well established and projected to continue. Two primary factors contributing to the nursing shortage are insufficient numbers of faculty and insufficient clinical sites for students. Innovative academic-service partnerships are realigning these scarce resources to improve the quality of clinical education and build cultures of safety. Relationships among students, staff nurses, faculty, and the institutions where they practice are central to students' socialization, professional role development, and transition to practice. Five recommendations to strengthen these professional relationships are suggested to: reenvision nursing student-staff nurse relationships, reconceptualize the clinical faculty role, enhance development for school-based faculty and staff nurses working with students, reexamine the depth and breadth of the clinical component, and strengthen the evidence for best practices in clinical nursing education. Five key outcomes are suggested to evaluate both traditional and emerging approaches to clinical nursing education. PMID- 19681536 TI - Promoting skill building and confidence in freshman nursing students with a "Skills-a-Thon". AB - Freshman nursing students returning for their second semester after summer break benefited by reviewing previously learned clinical skills presented in a Skills-a Thon. Skills stations were established and facilitated by faculty and senior students. Senior students were first trained in mentoring and specific steps in skills competencies. Freshman students demonstrated skills in various mock clinical situations including catheter insertion, sterile dressings, medications, and physical assessment. The strategy reinforced learning and provided an opportunity for students to experience risk-free skills performance among peers. Freshman students gained proficiency and appreciated guidance by senior students without the pressures of testing. Seniors benefited from a condensed version of the program to review their own skills prior to the event. Responses were positive, with students reporting improved performance and confidence with hands on application in a non-threatening environment. Nursing faculty observed improvement in skill performance and competence, and plan to offer future events. PMID- 19681537 TI - Turning simulation into reality: increasing student competence and confidence. AB - Clinical experiences are an essential part of nursing education as students learn technical skills, build on critical thinking skills, and hone skills in patient teaching. To build competence and confidence in each of these skill areas, an innovative clinical experience for senior students enrolled in women's health nursing was developed to provide nursing care and independent discharge teaching for postpartum mothers. Faculty facilitated this clinical experience by designing a simulation laboratory for students to practice their maternal self-care teaching and infant care skills prior to beginning their clinical rotation. In the hospital, students spent a day independently prioritizing new mothers' need for education and teaching new mothers to care for themselves and their newborns. Students reported confidence in teaching maternal self-care and newborn care, and satisfaction with this unique clinical experience. This approach may assist students in transferring skills learned in simulation laboratories to clinical practice. PMID- 19681538 TI - Teaching to the three apprenticeships: designing learning activities for professional practice in an undergraduate curriculum. AB - The Carnegie Foundation for the Advancement of Teaching recently completed a series of comparative studies that examined components and best practices in professional education and practice across five professions (clergy, law, medicine, nursing, and engineering). Across these disciplines, three apprenticeships were identified as necessary components of education for professional practice: an intellectual or cognitive apprenticeship, a skill-based apprenticeship related to clinical judgment and practice, and an apprenticeship to the ethical comportment or behavior of the profession. Although nursing education has a strong theory and clinical practice base historically, the comparative study of nursing education by the Carnegie Foundation found limited integration of the apprenticeships. Using an exemplar, this article discusses intentional design of learning objectives and activities to integrate learning across the three apprenticeships with an emphasis on key elements for professional practice in nursing. PMID- 19681540 TI - Thetis missed--dipping some master's athletes. PMID- 19681539 TI - Innovative learning activity. Evidence-based practice to outcomes management: spiraled learning activities, Part II. PMID- 19681541 TI - American health care system disaster. PMID- 19681543 TI - Arthroscopic triple-row rotator cuff repair: a modified suture-bridge technique. PMID- 19681544 TI - Pharmacological approaches for pediatric patients with osteomyelitis: current issues and answers. PMID- 19681545 TI - Techniques of obtaining and maintaining reduction during nailing of femur fractures. PMID- 19681546 TI - Lumbar disk herniation: what are reliable criterions indicative for surgery? PMID- 19681548 TI - Radiologic case study. Chondroblastoma with geographic bone destruction and a sclerotic rim. PMID- 19681557 TI - Home again: small houses for individuals with cognitive impairment. AB - The small house model of elder care emphasizes deinstitutionalization as a strategy to reduce the negative outcomes associated with nursing home care. The small house changes the philosophy, architecture, and organizational design of the institution and has been associated with higher quality of life and good quality of care. The intended benefits to individuals with cognitive impairment include better staff understanding of dementia care, improved physical environment, and safe, familiar patterns of everyday living. Initial research has been conducted, and additional research is underway to determine whether the outcomes match the intentions. PMID- 19681558 TI - When "no" means no: elderly patients' right to refuse treatment. AB - In the complicated health care and legal system of the United States, all patients are endowed with certain rights. The American Hospital Association and the U.S. Supreme Court have each delineated certain rights to which patients who have the competence and mental capacity to make decisions for themselves are entitled. One such right especially important to geriatric patients is the right to refuse treatment. Prior to treatment, nurses are often charged with obtaining the informed consent of the patient. To do so, the nurse must evaluate the patient's competency and mental capacity while explaining the proposed treatment, the benefits and risks involved, and other available treatment options, and then accept either the patient's voluntary assent or refusal. PMID- 19681559 TI - Psychotic events in Alzheimer's disease: application of the PLST model. AB - This article focuses on a review of the literature related to the known prevalence of psychotic events in individuals with Alzheimer's disease and associated aggressive, violent behavior toward family caregivers. It also describes the impact of behavioral disturbances on family caregivers and how use of the Progressively Lowered Stress Threshold model and nonpharmacological interventions cited in the literature can help manage these behaviors. Geriatric nurses armed with this information will be better prepared to provide caregivers with much-needed education to better understand psychotic events, as well as strategies to cope with associated behaviors. PMID- 19681561 TI - Cinemeducation: teaching end-of-life issues using feature films. AB - Given the rapidly escalating older adult population and the need to strengthen content on end-of-life care in nursing curricula, finding creative strategies to encourage student thinking about the dying process and personal choice in end-of life care is a timely challenge for nurse educators. There is an evolving body of literature in a variety of disciplines on cinemeducation, or the use of films to promote learning and personal awareness. This article describes an innovative assignment incorporating feature films designed not only to enhance student knowledge of the stages and process of grief and the complexity of human reactions when facing end-of-life situations, but also to engage students in thinking about the importance of quality of life in palliative care. The assignment was given to students enrolled in a second-semester, junior-level advanced adult health nursing course. Faculty and student evaluations of the assignment are discussed, and suggestions for selecting films and structuring such an assignment are proposed. PMID- 19681562 TI - Quality of life associated with adult day centers. AB - The purpose of this study was to describe the quality of life (QOL) of current adult day center (ADC) clients and their family caregivers in one region of Canada. Semi-structured interviews were conducted with 10 ADC clients and 10 caregivers. Data analysis used a modified constant comparative method. The major categories relating to QOL that emerged in the analysis included Physical Health and Well-Being, Social Networks/Relationships, Aging in Place, Safety, Respite, Activation, Respect and Inclusion, and Adequate Health Care Services. These themes are consistent with those of other studies of QOL of older adults. It was evident that ADCs played an important role in maintaining and improving the QOL of both the older adults and caregivers participating in this study. PMID- 19681566 TI - What nurses do. PMID- 19681563 TI - Effect of individualized music on agitation in individuals with dementia who live at home. AB - This pilot study investigated the effect of individualized music on agitation in individuals with dementia who live at home. Fifteen individuals listened to their preferred music for 30 minutes prior to peak agitation time, two times per week for 2 weeks, followed by no music intervention for 2 weeks. The process was repeated once. The findings showed that mean agitation levels were significantly lower while listening to music than before listening to the music. The findings of this pilot study suggest the importance of music intervention for individuals with dementia who live at home. PMID- 19681567 TI - The "connection" between continuing education and quality. AB - What is the connection between quality and continuing education? Are staff nurses able to identify that there is a connection? What are the implications for continuing education providers? This column describes one hospital's experience in connecting quality and continuing education. PMID- 19681568 TI - Treating ventricular tachycardia. AB - Ventricular tachycardia (VT) is a life-threatening cardiac dysrhythmia requiring emergency medical care. VT is readily recognized on the electrocardiogram. VT is usually caused by ischemic or structural heart disease, electrolyte disturbances, or the effects of drug therapy. Emergency treatment of VT follows the Advanced Cardiac Life Support (ACLS) algorithms for pulseless VT and stable and unstable VT with a pulse. PMID- 19681569 TI - Situations in which questions are valuable. AB - This column discusses several situations in which questions are valuable. PMID- 19681570 TI - Development of a competency framework for nurse managers in Ireland. AB - This article describes the results of an Irish national study on the Competency Model for Nursing Management commissioned by the Office of Health Management. More than 300 nurse managers and 80 service stakeholders (other professionals, managers, and service colleagues) participated in the development of generic competencies for nurse managers and specific competencies for three levels of managers-director level, middle manager level, and front-line managers. Examples of behavioral indicators (both positive and negative) for each competency level also have been delineated. Future efforts are being directed toward evaluating the usefulness of the competency model for assessing readiness to manage among job applicants, implementation of continuing education programs for nurse managers, and overall career development and planning. PMID- 19681572 TI - Implementation of the 6-week educational component in the Res-Care intervention: process and outcomes. AB - A 6-week restorative care educational program (30 minutes weekly) was conducted for nursing assistants. A total of 523 nursing assistants from 12 nursing homes were recruited: 265 were at treatment sites and 258 were at control sites. The mean age of the participants was 38.1 years (SD = 12.0). The majority were female (486; 93%) and African American (466; 89%). The nursing assistants had an average of 14.7 (SD = 3.8) years of education and 11.5 (SD = 8.6) years of experience. Control sites had a single 30-minute in-service on managing behavioral problems commonly associated with dementia. A total of 33% of the nursing assistants who consented to participate at the treatment sites attended all six classes, and 53% of those who did not attend at least three classes received one-on-one review of the class content. Overall, 86% of the nursing assistants who consented to participate attended the 6-week educational program. At the control sites, 18% of the nursing assistants who consented to participate attended the in-service training. There was a significant increase in restorative care knowledge (SD = 2.7, F = 280.4, p < .05) in treatment group participants. The techniques used in this intervention were effective in helping to expose nursing assistants to educational sessions and increase their knowledge of nursing care practices. PMID- 19681573 TI - Applied epidemiology for public health and community-based nurses. AB - As concerns mount surrounding exposure to environmental hazards and other public health threats, skills in surveillance and outbreak investigation are becoming critical competencies for public health and community-based nurses. This article describes a continuing education course designed in collaboration with a variety of stakeholders to address the need for epidemiological skills for rural nurses working in community-based settings. This project was funded through the Health Resources and Services Administration's Public Health Nursing Continuing Education program and was offered in both in-person and online formats. Topics, including applying epidemiological techniques to plan, implement, and evaluate public health programs, were taught using case studies and an array of online resources. Evaluation measures reflected improved understanding of epidemiological methods, a high level of satisfaction with the course and faculty, and strategies for applying these skills in professional nursing practice. PMID- 19681574 TI - Voices of experience: reflections of nurse educators. AB - BACKGROUND: Recent data reported by the American Association of Colleges of Nursing suggest that nursing education is experiencing a continued increase in applicants. However, because of a faculty shortage, many qualified students are denied enrollment. Strategies that would encourage individuals to consider a career in nursing education would somewhat alleviate applicant restrictions. METHODS: A descriptive phenomenological study was conducted exploring experienced baccalaureate educators' perspectives. Questions were posed about their role experiences. Analysis was conducted with van Manen's procedural steps for examination. RESULTS: Emergent themes were identified. These themes, reflective of the educators' experience, offer insight and guidance to novice educators striving for success. CONCLUSION: To alleviate the nursing shortage, increased efforts should be directed toward encouraging qualified individuals to pursue a career as a nurse educator and mentoring those who are novices. The wisdom gleaned from experienced nurse educators may help achieve these goals. PMID- 19681576 TI - Lessons learned while conducting a clinical trial to facilitate evidence-based practice: the neophyte researcher experience. AB - Health care is incredibly complex. Now, more than ever, health care providers are being called on to deliver care that is based on evidence and is consistent with current professional knowledge. Educators often struggle to find time when staff are available to participate in scheduled education. The work of nursing staff is to provide patient care. This article describes the journey of neophyte researchers who conducted a clinical trial as a strategy to implement evidence based practice. Although educational opportunities for the staff were included as part of the trial, there were also many challenges applicable to the neophyte researcher role. Those challenges included the participants' neophyte knowledge and perception of the research process, vendor relationships, data collection methods, staff perceptions, and the culture of the institution itself. PMID- 19681575 TI - Effect of an acute care geriatric educational program on fall rates and nurse work satisfaction. AB - BACKGROUND: A successful faculty-mentored geriatric nurse education program resulted from an acute care-academic partnership. PURPOSE: This study educated nurses in best geriatric nursing practices to enhance effective management of common geriatric problems as well as to improve nurse work satisfaction. METHODS: Forty-seven nurses completed an online course in geriatric best practices. RESULTS: Pre- and posttest comparisons showed a declining trend in the incidence of falls and improved satisfaction with autonomy, task requirements, and nurse-to nurse interaction among participants. Units in which two or more participants achieved national certification had significantly lower fall rates 3 months after program completion. Participants who attained national certification also had significantly improved satisfaction with professional status. CONCLUSION: This partnership represents a replicable model that facilitated the advancement of evidence-based practice and enhanced nurses' knowledge of best practices in caring for older adults. Early evidence shows that these successes may improve patient outcomes. PMID- 19681577 TI - Perovskite-type oxide thin film integrated fiber optic sensor for high temperature hydrogen measurement. AB - Small size fiber optic devices integrated with chemically sensitive photonic materials are emerging as a new class of high-performance optical chemical sensor that have the potential to meet many analytical challenges in future clean energy systems and environmental management. Here, we report the integration of a proton conducting perovskite oxide thin film with a long-period fiber grating (LPFG) device for high-temperature in situ measurement of bulk hydrogen in fossil- and biomass-derived syngas. The perovskite-type Sr(Ce(0.8)Zr(0.1))Y(0.1)O(2.95) (SCZY) nanocrystalline thin film is coated on the 125 microm diameter LPFG by a facile polymeric precursor route. This fiber optic sensor (FOS) operates by monitoring the LPFG resonant wavelength (lambda(R)), which is a function of the refractive index of the perovskite oxide overcoat. At high temperature, the types and population of the ionic and electronic defects in the SCZY structure depend on the surrounding hydrogen partial pressure. Thus, varying the H(2) concentration changes the SCZY film refractive index and light absorbing characteristics that in turn shifts the lambda(R) of the LPFG. The SCZY-coated LPFG sensor has been demonstrated for bulk hydrogen measurement at 500 degrees C for its sensitivity, stability/reversibility, and H(2)-selectivity over other relevant small gases including CO, CH(4), CO(2), H(2)O, and H(2)S, etc. PMID- 19681579 TI - Easy route to superhydrophobic copper-based wire-guided droplet microfluidic systems. AB - Droplet-based microfluidic systems are an expansion of the lab on a chip concept toward flexible, reconfigurable setups based on the modification and analysis of individual droplets. Superhydrophobic surfaces are one suitable candidate for the realization of droplet-based microfluidic systems as the high mobility of aqueous liquids on such surfaces offers possibilities to use novel or more efficient approaches to droplet movement. Here, copper-based superhydrophobic surfaces were produced either by the etching of polycrystalline copper samples along the grain boundaries using etchants common in the microelectronics industry, by electrodeposition of copper films with subsequent nanowire decoration based on thermal oxidization, or by a combination of both. The surfaces could be easily hydrophobized with thiol-modified fluorocarbons, after which the produced surfaces showed a water contact angle as high as 171 degrees +/- 2 degrees . As copper was chosen as the base material, established patterning techniques adopted from printed circuit board fabrication could be used to fabricate macrostructures on the surfaces with the intention to confine the droplets and, thus, to reduce the system's sensitivity to tilting and vibrations. A simple droplet-based microfluidic chip with inlets, outlets, sample storage, and mixing areas was produced. Wire guidance, a relatively new actuation method applicable to aqueous liquids on superhydrophobic surfaces, was applied to move the droplets. PMID- 19681578 TI - Antibody selection for immobilizing living bacteria. AB - We report a comparative study of the efficacy of immobilizing living bacteria by means of seven antibodies against bacterial surface antigens associated with Salmonella enterica Serovar Typhimurium. The targeted bacterial antigens were CFA/I fimbriae, flagella, lipopolysaccharides (LPS), and capsular F1 antigen. The best immobilization of S. Typhimurium was achieved with the antibody against CFA/I fimbriae. The immobilization of bacteria using antiflagellin showed significant enhancement if the flagella rotary motion was paralyzed. Of the four antibodies targeting LPS structures, only one, the antibody against the O-antigen polysaccharides, showed a relatively efficient bacterial immobilization. No bacterial immobilization was achieved using the antibody against F1 antigen, presumably because F1 protein can detach from the bacterial surface easily. The results suggest that an antibody for bacterial immunoimmobilization should target a surface antigen which extends out from the bacterial surface and is tightly attached to the bacterial cell wall. The microarrays of living S. Typhimurium cells immobilized in this manner remained viable and effective for at least 2 weeks in growth medium before a thick biofilm covered the whole surface. PMID- 19681580 TI - Silicon powder: the first nonmetal elemental catalyst for aminobromination of olefins with TsNH(2) and NBS. AB - Silicon powder was found, for the first time, to be an efficient alternative to transition metal catalysts for aminobromination of alpha,beta-unsaturated carbonyl compounds and simple olefins with p-toluenesulfonamide (4-TsNH(2)) and NBS, affording the aminobrominated products in high yields and regio- and stereoselectivity. The high reactivity of electron-rich substrates reveals that the reaction has the electrophilic addition feature. PMID- 19681581 TI - Bent ladder-type hexaphenylene with carbazole core and spiro linkage as stable and efficient blue emitter. AB - A bent ladder-type hexaphenylene with a carbazole core and spiro-linkage is designed and synthesized by using the ortho-linked spirobifluorene. The design eliminates the possibility of forming a positional isomer. As a blue-emitter, the BLHPC shows good thermal and color stability. A simple light-emitting device fabricated from BLHPC exhibits a maximum current efficiency of 1.46 cd/A and a maximum luminance of 505 cd/m(2). PMID- 19681582 TI - Ab initio study of the electron transfer in an ionized stacked complex of guanines. AB - The charge transfer process in an ionized stacking of two consecutive guanines (G(5')G(3'))(+) has been studied by means of state-averaged CASSCF/MRCI and RASSCF/RASPT2 calculations. The ground and two first excited states of the radical cation have been characterized, and the topology of the corresponding potential energy surfaces (PESs) has been studied as a function of all intermolecular geometrical parameters. The results demonstrate that the charge transfer process in (G(5')G(3'))(+) is governed by the avoiding crossing between the ground and first excited states of the complex. Relative translation motions of both guanines in their molecular planes are shown to lead to the charge migration between G(5') and G(3'). Five stationary points (three minima and two saddle points) have been characterized along the reaction path describing the passage of the positive charge from G(5') to G(3'). The global minimum on the PES is found to correspond to the charge configuration G(5')(+)G(3'). The existence of an intermediate minimum along the reaction path has been established, characterizing a structure where the positive charge is equally distributed between the two guanines. The calculated energy profile allowed us to determine the height of the potential energy barrier (7.33 kcal/mol) and to evaluate the electronic coupling at a geometry close to the avoiding crossing (3.6 kcal/mol). Test calculations showed that the topology of the ground state PES of the complex GG(+) is qualitatively conserved upon optimization of the intramolecular geometrical parameters of the stationary points. PMID- 19681583 TI - The effect of temperature and dot size on the spectral properties of colloidal InP/ZnS core-shell quantum dots. AB - Visual color changes between 300 and 510 K were observed in the photoluminescence (PL) of colloidal InP/ZnS core-shell nanocrystals. A subsequent study of PL spectra in the range 2-510 K and fitting the temperature dependent line shift and line width to theoretical models show that the dominant (dephasing) interaction is due to scattering by acoustic phonons of about 23 meV. Low temperature photoluminescence excitation measurements show that the excitonic band gap depends approximately inversely linearly on the quantum dot size d, which is distinctly weaker than the dependence predicted by current theories. PMID- 19681585 TI - Structure of high internal phase aqueous-in-oil emulsions and related inverse micelle solutions. 4. Surfactant mixtures. AB - The effects of combinations of surfactants on the structure and stability of high internal phase water-in-hexadecane and saturated ammonium nitrate-in-hexadecane oil-based emulsions and oil-based inverse micellar solutions are reported. The combinations were 750, 1,200, and 1,700 molecular weight monodisperse and 450 and 1,000 molecular weight polydisperse polyisobutylene acid amides, and sorbitan monooleate. The samples made from mixtures have qualitatively similar nanostructures to emulsions made from single surfactants. Again, for the emulsions, micrometer-scale aqueous droplets are dispersed in a continuous oil phase, which contains inverse spherical micelles composed of surfactant, hexadecane, and water. In quantitative terms, lower average surfactant molecular weight, lower ammonium nitrate content, and lower surfactant content increased the swelling of micelles, their water content, and the tendency of the emulsion to be unstable and form a sponge phase. This instability also allows micelle plasticity such that their geometry and content in mixed surfactant systems are not simply predictable by interpolation from single surfactant systems. An example was found of a mixed micelle 3 times larger than either single component micelle. The observed behavior suggests that mixing surfactant molecules of very different molecular weights destabilizes the emulsions, while mixing surfactants close in molecular weight has the opposite effect. The synergistic effects of surfactant molecular weight polydispersity and binary mixing are most marked for 1:1 molecular mixtures of surfactant. PMID- 19681584 TI - Hydrophobic solvation: a 2D IR spectroscopic inquest. AB - For decades, the enigma of the hydrophobic force has captured the imagination of scientists. In particular, Frank and Evans' idea that the hydrophobic effect was mainly due to some kind of "iceberg" formation around a hydrophobic solute stimulated many experiments and molecular dynamics simulation studies. A better understanding of hydrophobic interactions will aid understanding in many contexts including protein structural dynamics and functioning in biological systems. In this Account, we present results of two-dimensional infrared (2D IR) spectroscopy experiments on the OH-stretch vibrational mode of water molecules near hydrophobic groups in concentrated solutions with tetramethylurea (TMU). The frequency of the OH vibration is a sensitive probe for environmental dynamics and, in particular, for the strength of the hydrogen bond. Two-dimensional IR spectroscopy can trace time correlations of the vibrational frequency at the scale of hundreds of femtoseconds and thus provides valuable insight into the effect of hydrophobic solutes on the dynamics of a hydrogen-bond network. We compare the 2D spectroscopic results with molecular dynamics (MD) simulations to obtain a microscopic picture of hydrophobic solvation. We observe two different types of hydrogen-bond dynamics in the water/TMU mixtures. We attribute the "fast" ( approximately 100 fs) dynamics to highly coordinated water molecular jump reorientations and assigned the "slow" (>1 ps) dynamics to water translational motions that are strongly suppressed by the TMU molecules. Molecular dynamics simulations demonstrate a clear correlation between the slowed dynamics and the translational mobility of water. This finding indicates that the molecular-jump reorientations are switched off near hydrophobic groups. The fifth water molecule, which is required to form a defect state in the tetrahedral surroundings, cannot approach the hydrogen-bonded pair to initiate the molecular jump. As a result, the rate of the jumping events sharply decreases, which, in turn, strongly slows the rotation of the water molecules. Our findings suggest that water molecules in the hydrophobic solvation shell do not exhibit an increased tetrahedral ordering compared with the bulk but that the hydrogen-bond dynamics in the two cases are different. This result also indicates that consideration of a hydrogen bond's dynamics could be critical for its definition. PMID- 19681586 TI - Structure of high internal phase aqueous-in-oil emulsions and related inverse micelle solutions. 3. Variation of surfactant. AB - The small angle neutron scattering from high internal phase water-in-hexadecane and saturated ammonium nitrate-in-hexadecane emulsions is compared with that from related hexadecane-based inverse micellar solutions. Three molecular weights of the monodisperse polyisobutylene acid amide (PIBSA) surfactant 750, 1200, and 1700 were studied over a range of surfactant concentrations. As an additional comparison, emulsions based on sorbitan monooleate and isostearate surfactants were investigated. The scattering from molecular weight 1200 water-based PIBSA emulsions can be fitted at all concentrations to a model with a surfactant coated aqueous droplet-oil interface together with the majority of the surfactant in the oil phase of the emulsion in the form of inverse micelles. Variation of the molecular weight shows a variety of phases of increasing curvature: lamellar, sponge, and, most commonly, the emulsion structure described above. In addition, the molecular weight affects the oil component in the emulsions, which can contain either cylindrical micelles or spherical micelles of varying water but constant hexadecane content. Increased phase curvature is favored by both increased PIBSA molecular weight and ammonium nitrate dissolved in the water. These observations are consistent with "Wedge theory". The structures observed in the emulsions are close to those observed in related inverse micellar solutions made from hexadecane, the surfactant, and water. Lower concentrations of surfactant in the micellar solutions decrease micelle curvature, except where the inverse micelles are spherical and small; here, there is little effect of dilution. Substitution of sorbitan surfactants for PIBSAs produces slightly less organized but similar structures, with smaller spherical micelles containing proportionally more water. The aqueous-oil droplet interface has a relatively invariant monolayer of adsorbed surfactant. For all emulsions, we can infer from the mass balance that micelle concentrations are depressed in the inverse micellar solutions because up to half the added surfactant is present as individually dissolved molecules. PMID- 19681587 TI - Bayesian single-exponential kinetics in single-molecule experiments and simulations. AB - In this work, we develop a fully Bayesian method for the calculation of probability distributions of single-exponential rates for any single-molecule process. These distributions can even be derived when no transitions from one state to another have been observed, since in that case the data can be used to estimate a lower bound on the rate. Using a Bayesian hypothesis test, one can easily test whether a transition occurs at the same rate or at different rates in two data sets. We illustrate these methods with molecular dynamics simulations of the folding of a beta-sheet protein. However, the theory presented here can be used on any data from simulation or experiment for which a two-state description is appropriate. PMID- 19681588 TI - A Kirkwood-Buff derived force field for thiols, sulfides, and disulfides. AB - A force field has been developed for molecular simulations of methanethiol, dimethyl sulfide, and dimethyl disulfide mixtures. The force field specifically attempts to balance the solvation and self-association of these solutes in solution mixtures with methanol. The force field is based on the Kirkwood-Buff (KB) theory of solutions and is parametrized using the KB integrals obtained from the experimental activity coefficients for the solution mixtures. The transferability of the force field was tested and confirmed by the accurate prediction of the activity coefficients for methanethiol/dimethyl sulfide solutions, which were not used in the initial parametrization of the force fields. The ideality of this latter solution is excellently reproduced. The applicability of the force field to simulations in water was corroborated with a reasonably accurate prediction for the low solubility of dimethyl sulfide in water. The aggregation of methanol molecules at low methanol mole fractions displayed by all the mixtures is reproduced and further analyzed. PMID- 19681589 TI - Guttiferones O and P, prenylated benzophenone MAPKAPK-2 inhibitors from Garcinia solomonensis. AB - Two prenylated benzophenones, guttiferones O (1) and P (2), were isolated from the stem bark of the Papua New Guinean plant Garcina solomonensis. The structures of these compounds and their relative configurations were determined by spectroscopic methods. Both compounds inhibited the phosphorylation of the synthetic biotinylated peptide substrate KKLNRTLSVA by the serine/threonine protein kinase MAPKAPK-2 with IC(50) values of 22.0 microM. PMID- 19681590 TI - Metal-ligand bonds of second- and third-row d-block metals characterized by density functional theory. AB - This paper presents systematic data for 200 neutral diatomic molecules ML (M is a second- or third-row d-block metal and L = H, F, Cl, Br, I, C, N, O, S, or Se) computed with the density functionals TPSSh and BP86. With experimental structures and bond enthalpies available for many of these molecules, the computations first document the high accuracy of TPSSh, giving metal-ligand bond lengths with a mean absolute error of approximately 0.01 A for the second row and 0.03 A for the third row. TPSSh provides metal-ligand bond enthalpies with mean absolute errors of 37 and 44 kJ/mol for the second- and third-row molecules, respectively. Pathological cases (e.g., HgC and HgN) have errors of up to 155 kJ/mol, more than thrice the mean (observed with both functionals). Importantly, the systematic error component is negligible as measured by a coefficient of the linear regression line of 0.99. Equally important, TPSSh provides uniform accuracy across all three rows of the d-block, which is unprecedented and due to the 10% exact exchange, which is close to optimal for the d-block as a whole. This work provides an accurate and systematic prediction of electronic ground state spins, characteristic metal-ligand bond lengths, and bond enthalpies for many as yet uncharacterized diatomics, of interest to researchers in the field of second- and third-row d-block chemistry. We stress that the success of TPSSh cannot be naively extrapolated to other special situations such as, e.g., metal metal bonds. The high accuracy of the procedure further implies that the effective core functions used to model relativistic effects are necessary and sufficient for obtaining accurate geometries and bond enthalpies of second- and third-row molecular systems. PMID- 19681591 TI - Enantioselective synthesis of beta-trifluoromethyl-beta-lactones via NHC catalyzed ketene-ketone cycloaddition reactions. AB - The highly diastereo- and enantioselective synthesis of beta-trifluoromethyl-beta lactones bearing two contiguous stereocenters was realized by chiral N heterocyclic carbene-catalyzed formal cycloaddition reaction of alkyl(aryl)ketenes and trifluoromethyl ketones. PMID- 19681592 TI - Silylene-mediated ring contraction of homoallylic ethers to form allylic silanes. AB - (-)-Isopulegol derivatives undergo a ring contraction under silylene-mediated conditions to provide cyclopentane products. Silylene transfer to other homoallylic ethers did not provide the ring contraction products. Allylic silane products were elaborated to determine the stereochemical course of the ring contraction reaction. A mechanism for the transformation is proposed. PMID- 19681594 TI - Additional supra-self-assembly of human serum albumin under amyloid-like-forming solution conditions. AB - Protein aggregation has a multitude of consequences ranging from affecting protein expression to its implication in different diseases. Of recent interest is the specific form of aggregation leading to the formation of amyloid fibrils, structures associated with diseases such as Alzheimer's disease. These fibrils can further associate in other more complex structures such as fibrillar gels, plaques, or spherulitic structures. In the present work, we describe the physical and structural properties of additional supraself-assembled structures of human serum albumin under solution conditions in which amyloid-like fibrils are formed. We have detected the formation of ordered aggregates of amyloid fibrils, i.e., spherulites which possess a radial arrangement of the fibrils around a disorganized protein core and sizes of several micrometers by means of polarized optical microscopy, laser confocal microscopy, and transmission electron microscopy. These spherulites are detected both in solution and embedded in an isotropic matrix of fibrillar gels. In this regard, we have also noted the formation of protein gels when the protein concentration and/or ionic strength exceds a threshold value (the gelation point) as observed by rheometry. Fibrillar gels are formed through intermolecular nonspecific association of amyloid fibrils at a pH far away from the isolectric point of the protein where protein molecules seem to display a "solid-like" behavior due to the existence of non-DLVO (Derjaguin-Landau-Verwey-Overbeck) intermolecular repulsive forces. As the solution ionic strength increases, a coarsening of this type of gel is observed by environmental scanning microscopy. In contrast, at pH close to the protein isoelectric point, particulate gels are formed due to a faster aggregation process, which does not allow substantial structural reorganization to enable the formation of ordered structures. This behavior also additionally corroborates that the existence of particulates might also be a generic property of all polypeptide chains as amyloid fibril formation under suitable conditions. PMID- 19681593 TI - N-terminal aliphatic residues dictate the structure, stability, assembly, and small molecule binding of the coiled-coil region of cartilage oligomeric matrix protein. AB - The coiled-coil domain of cartilage oligomeric matrix protein (COMPcc) assembles into a homopentamer that naturally recognizes the small molecule 1,25 dihydroxyvitamin D(3) (vit D). To identify the residues critical for the structure, stability, oligomerization, and binding to vit D as well as two other small molecules, all-trans-retinol (ATR) and curcumin (CCM), here we perform an alanine scanning mutagenesis study. Ten residues lining the hydrophobic pocket of COMPcc were mutated into alanine; of the mutated residues, the N-terminal aliphatic residues L37, L44, V47, and L51 are responsible for maintaining the structure and function. Furthermore, two polar residues, T40 and Q54, within the N-terminal region when converted into alanine improve the alpha-helical structure, stability, and self-assembly behavior. Helical stability, oligomerization, and binding appear to be linked in a manner in which mutations that abolish helical structure and assembly bind poorly to vit D, ATR, and CCM. These results provide not only insight into COMPcc and its functional role but also useful guidelines for the design of stable, pentameric coiled-coils capable of selectively storing and delivering various small molecules. PMID- 19681595 TI - Simultaneous electrochemical determination of glutathione and glutathione disulfide at a nanoscale copper hydroxide composite carbon ionic liquid electrode. AB - Direct simultaneous electrochemical determination of glutathione (GSH) and glutathione disulfide (GSSG) has been presented using a nanoscale copper hydroxide carbon ionic liquid composite electrode. To the best of our knowledge, this is the first report on the simultaneous determination of these two biologically important compounds based on their direct electrochemical oxidation. Incorporation of copper(II) hydroxide nanostructures in the composite electrode results in complexation of Cu(II) with the thiol group of GSH and leads to a significant decrease in GSH oxidation overpotential, while an anodic peak corresponding to the direct oxidation of GSSG as the product of GSH oxidation is observed at higher overvoltages. Low detection limits of 30 nM for GSH and 15 nM for GSSG were achieved based on a signal-to-noise ratio of 3. The proposed method is free from interference of cysteine, homocysteine, ascorbic acid (AA), and uric acid (UA). No electrode surface fouling was observed during successive scans. Stability, high sensitivity, and low detection limits made the proposed electrode applicable for the analysis of biological fluids. PMID- 19681596 TI - Surrogate H/D detection strategy for protein conformational analysis using MS/MS data. AB - Amide hydrogen/deuterium exchange (H/DX) measurements by mass spectrometry provide a powerful tool for probing the structure and dynamics of proteins. In order to extend such measurements to complex multiprotein systems, new methods for delivering higher sensitivity and sequence coverage are required. In this study, we investigated the utility of tandem mass spectrometry (MS/MS) for providing an alternative to the conventional MS mode of operation applied to bottom-up H/DX experiments. Specifically, we aimed to determine whether differential deuteration measurements of collisionally induced dissociation (CID) generated product ions can serve as effective surrogates for their corresponding intact peptide, thus providing an additional dimension for analysis. Replicate deuterium measurements of calmodulin (in its apo and holo forms) were obtained from peptic digests in both the MS and MS/MS domains and studied as a function of % deuteration, fragment ion selection, and contaminant level. We show that successful acquisition of MS/MS data for deuterated peptides requires controlled expansion of the isotopic envelope by limiting the range of deuterium label applied in the exchange-in reaction (< or = 50%) and that automation of ion selection via data-dependent acquisition is ultimately dependent upon peak detection algorithms. Upon full transmission of the isotopic envelope, fragment data demonstrate that all ions, with the exception of neutral loss fragment ions, return deuteration ratios reflecting the apo/holo transition that are in general agreement with values obtained from the corresponding precursor ions. The agreement is limited primarily by the ion statistics for each fragment, as the base peak in the MS/MS spectra provided the best correlation regardless of its m/z. We highlight that the freedom to select the base peak as a surrogate for the precursor ion derives from extensive H/D scrambling inducible under conventional CID conditions. When spectral interference prohibits conventional H/DX-MS measurements, we further show that the surrogate approach recovers accurate and precise per-peptide deuteration levels. Thus, a generalized strategy is presented in which CID-based automated H/DX-MS/MS acquisition can be used to extend measurements to complex protein systems, exceeding the peptide capacity of conventional H/DX-MS alone. PMID- 19681597 TI - Loss of Toll-like receptor 2 and 4 leads to differential induction of endoplasmic reticulum stress and proapoptotic responses in the intestinal epithelium under conditions of chronic inflammation. AB - Toll-like receptors (TLRs) play an important role in the recognition of microbial molecular patterns of infectious and commensal bacteria and their expression in various tissues including the intestinal epithelium orchestration of the innate and adaptive immune defense mechanisms. Changes in the TLR signaling pathways due to host genetic predispositions may turn a physiological response into a pathological situation including failure of bacterial clearance and development of chronic inflammation. The aim of this study was to characterize the role of TLR2 or TLR4 deficiency in epithelial cell stress responses under noninflamed and inflamed conditions using TLR-deficient mice and TLR(-/-) cross-bred IL-10 deficient mice as a model for genetically driven experimental colitis. Primary intestinal epithelial cells (IEC) were isolated from specific-pathogen-free wild type, TLR2-, TLR4-, IL-10-, IL-10XTLR2- and IL-10XTLR4-deficient mice at the age of 1, 8, and 16 weeks. Histopathological analysis showed absence of tissue pathology (score 0-12) in distal colon sections of TLR2- and TLR4-deficient mice. In addition, TLR2- but not TLR4-deficient mice cross-bred to the IL-10-deficient background develop moderate colitis, suggesting different effects of these pattern recognition receptors in regulating disease mechanisms. Proteome analysis revealed significantly regulated proteins associated with endoplasmic reticulum (ER) and mitochondrial stress responses in the epithelium. In contrast to TLR2(-/ ) and IL-10XTLR2(-/-) mice, the induction of the ER-associated chaperone grp-78 was dissociated from the activation of proapoptotic caspase 3 cleavage in noninflamed TLR4(-/-) and IL10XTLR4(-/-) mice. These results suggest that ER associated cellular stress responses play an important role in epithelial cells homeostasis leading to beneficial but also deleterious effects. We hypothesize that ER stress-associated processes in the absence of TLR2 and TLR4 differentially affect host responses and epithelial functions under conditions of genetically driven chronic intestinal inflammation. PMID- 19681598 TI - Surface coating directed cellular delivery of TAT-functionalized quantum dots. AB - TAT peptide functionalized shell-core ZnS-CdSe quantum dots (QDs) have been prepared by three different methods, direct ligand exchange with cysteine terminated TAT (TAT-QD(lig exch)), and covalent conjugation to QDs coated with silanes (TAT-QD(silica)) and polyacrylate derivatives (TAT-QD(polyacrylate)). The silica and polyacrylate coatings incorporated multiple primary and secondary amines, introducing positive surface charges onto the QDs, providing high water solubility and sites for peptide conjugation, while inducing the "proton sponge effect". The different coating methods produced particles of different sizes, surface charges, and colloidal stability; these factors jointly influenced the cellular uptake and subcellular localization of these particles. As the particle size increased, (TAT-QD(lig exch) (6 nm) < TAT-QD(silica) (10 nm) < QD(polyacrylate) (25 nm)), both the particle surface charge and cellular uptake increased. The smaller TAT-QD(lig exch) and TAT-QD(silica) particles were localized mainly in the perinuclear regions, while the larger TAT QD(polyacrylate) particles were localized in both the perinuclear regions and the lysosomes. Compared to the other TAT-QDs, TAT-QD(lig-exch) has a lower colloidal stability and was more cytotoxic due to the weak binding of the ligands. PMID- 19681599 TI - Exploring damage recognition models in prokaryotic nucleotide excision repair with a benzo[a]pyrene-derived lesion in UvrB. AB - The UvrB protein is a central unit for damage recognition in the prokaryotic nucleotide excision repair system, which excises bulky DNA lesions. We have utilized molecular modeling and MD simulations based on crystal structures, mutagenesis, and fluorescence data, to model the 10R-(+)-cis-anti-B[a]P-N2-dG lesion, derived from the tumorigenic (+)-anti-B[a]PDE metabolite of benzo[a]pyrene, at different locations on the inner and outer strand in UvrB. Our results suggest that this lesion is accommodated on the inner strand where it might translocate through the tunnel created by the beta-hairpin and UvrB domain 1B and ultimately could be housed in the pocket behind the beta-hairpin prior to excision by UvrC. Lesions that vary in size and shape may be stopped at the gate to the tunnel, within the tunnel, or in the pocket when UvrC initiates excision. Common features of beta-hairpin intrusion between the two DNA strands and nucleotide flipping manifested in structures of prokaryotic and eukaryotic NER lesion recognition proteins are consistent with common recognition mechanisms, based on lesion-induced local thermodynamic distortion/destabilization and nucleotide flipping. PMID- 19681601 TI - Near-infrared continuous-wave light driving a two-photon photochromic reaction with the assistance of localized surface plasmon. AB - We demonstrate that a photochromic reaction can be driven by irradiation from a weak, near-infrared continuous-wave (NIR-CW) laser light. A two-photon ring opening photochromic reaction of a diarylethene (DE) derivative can be induced by irradiation with a NIR-CW laser light (lambda = 808 nm). An ultrathin polymer film doped with DE in its closed form was coated onto a gold-nanoparticle integrated glass substrate. Upon irradiation of the sample with a CW laser at low fluence (0.1-4.0 W/cm(2)), we could clearly observe bleaching of the DE (ring opening reaction). Following the IR irradiation, the bleached absorption could be reversibly recovered by applying UV irradiation (ring-closing reaction). We verified that the yield of the photochromic ring-opening reaction of the DE was proportional to the square of the irradiation fluence. The origin of this NIR-CW induced two-photon photochromic reaction is an "enhancing effect" that acts on the electromagnetic field (localized surface plasmon) of the gold nanoparticles. The DE interacts with the surface plasmon and receives energy from two photons, which excites it to a state from which the ring-opening reaction can be initiated. PMID- 19681602 TI - High oxidation state rhodium and iridium bis(silyl)dihydride complexes supported by a chelating pyridyl-pyrrolide ligand. AB - New rhodium and iridium complexes containing the bidentate ligand 3,5-diphenyl-2 (2-pyridyl)pyrrolide (PyPyr) were prepared. The bis(ethylene) complex (PyPyr)Rh(C(2)H(4))(2) (3) reacted with HSiEt(3), HSiPh(3), and HSi(t)BuPh(2) to produce the 16-electron Rh(V) bis(silyl)dihydrides (PyPyr)Rh(H)(2)(SiEt(3))(2) (8), (PyPyr)Rh(H)(2)(SiPh(3))(2) (9), and (PyPyr)Rh(H)(2)(Si(t)BuPh(2))(2) (10), respectively. The analogous Ir(V) bis(silyl)dihyride (PyPyr)Ir(H)(2)(SiPh(3))(2) (11) has also been synthesized. X-ray crystallography reveals that 9-11 adopt a coordination geometry best described as a bicapped tetrahedron. Silane elimination from 9 and 10 occurred in the presence of either HSiEt(3) or PPh(3). Mechanistic studies of the silane exchange process involving 10 and free HSiEt(3) (to give 8) indicate that this process occurs by rate-limiting reductive elimination of HSi(t)BuPh(2) from 10 to generate a 14-electron Rh(III) intermediate of the type (PyPyr)Rh(H)(Si(t)BuPh(2)). PMID- 19681600 TI - Effect of thioredoxin deletion and p53 cysteine replacement on human p53 activity in wild-type and thioredoxin reductase null yeast. AB - Reporter gene transactivation by human p53 is inhibited in budding yeast lacking the TRR1 gene encoding thioredoxin reductase. To investigate the role of thioredoxin in controlling p53 activity, the level of reporter gene transactivation by p53 was determined in yeast lacking the TRX1 and TRX2 genes encoding cytosolic thioredoxin. Surprisingly, p53 activity was unimpaired in yeast lacking thioredoxin. Subsequent analyses showed that thioredoxin deletion suppressed the inhibitory effect of thioredoxin reductase deletion, suggesting that accumulation of oxidized thioredoxin in mutant yeast was necessary for p53 inhibition. Purified human thioredoxin and p53 interacted in vitro (Kd = 0.9 microM thioredoxin). To test the idea that dithio-disulfide exchange reactions between p53 and thioredoxin were responsible for p53 inhibition in mutant yeast, each p53 cysteine was changed to serine, and the effect of the substitution on p53 activity in TRR1 and Deltatrr1 yeast was determined. Substitutions at Zn coordinating cysteines C176, C238, or C242 resulted in p53 inactivation. Unexpectedly, substitution at cysteine C275 also inactivated p53, which was the first evidence for a non-zinc-coordinating cysteine being essential for p53 function. Cysteine substitutions at six positions (C124, C135, C141, C182, C229, and C277) neither inactivated p53 nor relieved the requirement for thioredoxin reductase. Furthermore, no tested combination of these six cysteine substitutions relieved thioredoxin reductase dependence. The results suggested that p53 dependence on thioredoxin reductase either was indirect, perhaps mediated by an upstream activator of p53, or was due to oxidation of one or more of the four essential cysteines. PMID- 19681603 TI - Supramolecular assembly of electrostatically stabilized, hydroxyproline-lacking collagen-mimetic peptides. AB - The mechanical and biological functions of the native collagens remain an inspiration in materials design, but widespread application of de novo collagens has been limited in part by the need for hydroxylated proline in the formation of stable triple helical structures. To address this continued need and to expand the potential for recombinant expression of functional, hydroxyproline-lacking collagen-mimetic peptides, we have designed a hydrophilic, nonrepetitive, and thermally stable collagen-mimetic peptide via the incorporation of triple-helix stabilizing charged triplets. The peptide sequence is also equipped with a type III-collagen-mimetic cystine knot at the C-terminus to facilitate covalent cross linking of the triple helix via simple air oxidation. Circular dichroic spectroscopy (CD) studies of this collagen-mimetic peptide revealed a typical, thermally stable, collagen triple helix signature with a weak positive maximum at 225 nm and a triple helix melting temperature (T(m)) of 35 and 43 degrees C for the reduced and oxidized forms, respectively. The thermal behavior was confirmed via analysis by differential scanning calorimetry. Interestingly, this hydroxyproline-lacking, collagen-mimetic peptide also assembles into nanorods and microfibrillar structures as observed via transmission electron microscopy. The identification and demonstrated useful collagen-mimetic properties of this peptide suggests important opportunities in the recombinant design of new collagen-based biomaterials. PMID- 19681604 TI - Antimicrobial properties of nanostructured hydrogel webs containing silver. AB - The need exists for biomaterials that prevent biofilm formation and associated infections. In this report, we have studied the synthesis, processing, and antimicrobial behavior of new silver-containing thermoplastic hydrogel nanofibrous webs. Thermoplastic hydrogels were synthesized from multiblock PEG POSS polyurethanes (PEG: poly(ethylene gylcol); POSS: polyhedral oligosilsesquioxane) and electrospun into nanofibrous webs (diameter approximately 150 nm), with or without AgNO(3). The nanofibrous hydrogels exhibited unusual shrinkage during water uptake, yielding a uniquely dense structure compared to hydrogels prepared from cast films. Antimicrobial activity was examined using exposure to Escherichia coli with daily refreshment of medium and inoculation to a controlled cell density. Nanofibrous hydrogels without silver featured the most rapid and most extensive biofilm formation, while the silver-containing nanofibrous hydrogel featured outstanding biofilm resistance, with biofilm formation taking hold only after 14 days of incubation in daily refreshed bacterial cultures. We envision application of the unique antimicrobial hydrogels as wound dressings that combine sustained bactericidal properties and lack of volumetric swelling during water uptake. PMID- 19681605 TI - Identification, biosynthesis, and characterization of polyhydroxyalkanoate copolymer consisting of 3-hydroxybutyrate and 3-hydroxy-4-methylvalerate. AB - In this study, we found that Ralstonia eutropha strain PHB(-)4 expressing the polyhydroxyalkanoate (PHA) synthase 1 (PhaC1(Ps)) from Pseudomonas sp. 61-3 synthesized a PHA copolymer containing a 3-hydroxybutyrate (3HB) and small amounts of 3-hydroxy-4-methylvalerate (3H4MV) and 3-hydroxyvalerate (3HV) from fructose as a sole carbon source. 3H4MV is a monomer unit that has hitherto not been reported as a naturally occurring component of PHAs. To increase the 3H4MV fraction in PHA copolymers, the culture medium was supplemented with four structural analogs that served as 3H4MV precursors. Of these, 4-methylvalerate (4MV) was the most efficient in increasing the 3H4MV content of PHA. The R. eutropha strain PHB(-)4 expressing Aeromonas cavaie PHA synthase (PhaC(Ac)) and its mutant (PhaC(Ac) NSDG mutant) were also able to synthesize 3H4MV-containing PHA from 4MV, increasing 3H4MV molar fraction in PHA copolymer up to 38 mol %. The structure and physical properties of P(3HB-co-3H4MV) copolymers were characterized by gel permeation chromatography, NMR spectroscopy, differential scanning calorimetry, and tensile testing. It can be shown that P(3HB-co-3H4MV) samples synthesized by PhaC(Ac) and its mutant NSDG were of higher molecular weights than those by PhaC1(Ps) and were a flexible and ductile material with moderate toughness. Additionally, the ductility can be kept for at least 180 days without significant deterioration caused by secondary crystallization when 3H4MV molar fraction was higher than 14 mol %. The newly identified 3H4MV unit is a promising monomer for improving material property and stability of 3HB-based copolymers. PMID- 19681606 TI - Helices, chirality and interpenetration: the versatility and remarkable interconversion of silver-copper cyanide frameworks. AB - The structural transformations between cesium silver-copper cyanides under modest conditions, both in solution and in the solid state, are described. Three new cesium silver(I) copper(I) cyanides with three-dimensional (3-D) framework structures were prepared as single crystals from a one-pot reaction initially heated under hydrothermal conditions. The first product to appear, Cs(3)Ag(2)Cu(3)(CN)(8) (I), when left in contact with the supernatant produced CsAgCu(CN)(3) (II) and CsAgCu(CN)(3).1/3H(2)O (III) over a few months via a series of thermodynamically controlled cascade reactions. Crystals of the hydrate (III) can be dehydrated to polycrystalline CsAgCu(CN)(3) (II) on heating at 100 degrees C in a remarkable solid-state transformation involving substantial breaking and reconnection of metal-cyanide linkages. Astonishingly, the conversion between the two known polymorphs of CsAg(2)Cu(CN)(4), which also involves a major change in connectivity and topology, occurs at 180 degrees C as a single-crystal to single-crystal transformation. Structural features of note in these materials include the presence of helical copper-cyanide chains in (I) and (II), which in the latter compound produce a chiral material. In (II) and (III), the silver-copper cyanide networks are both self- and interpenetrating, features also seen in the known polymorphs of CsAg(2)Cu(CN)(4). PMID- 19681608 TI - A pillared-layer coordination polymer with a rotatable pillar acting as a molecular gate for guest molecules. AB - The design of pore properties utilizing flexible motifs and functional groups is of importance to obtain porous coordination polymers with desirable functions. We have prepared a 3D pillared-layer coordination polymer, {[Cd(2)(pzdc)(2)L(H(2)O)(2)].5(H(2)O).(CH(3)CH(2)OH)}(n) (1, H(2)pzdc = 2,3 pyrazinedicarboxylic acid; L = 2,5-bis(2-hydroxyethoxy)-1,4-bis(4 pyridyl)benzene) showing (i) a rotatable pillar bearing ethylene glycol side chains acting as a molecular gate with locking/unlocking interactions triggered by guest inclusion between the side chains, (ii) framework flexibility with slippage of the layers, and (iii) coordinatively unsaturated metal centers as guest accessible sites through the removal of the water coligands. The framework clearly shows reversible single-crystal-to-single-crystal transformations in response to the removal and rebinding of guest molecules, the observation of these processes has provided fundamental clues to the understanding of the sorption profiles. The X-ray structures indicate that the 3D host framework is retained during the transformations, involving mainly rotation of the pillars and slippage of the layers. The structure of dried form 2, [Cd(2)(pzdc)(2)L](n), has no void volume and no water coligands. Interestingly, the adsorption isotherm of water for 2 at 298 K exhibits three distinct steps coinciding with the framework functions. Compound 2 favors the uptake of CO(2) (195 K) over N(2) (77 K) and O(2) (77 K). Above all, we report on a molecular gate with a rotational module exhibiting a locking/unlocking system which accounts for gate-opening type sorption profiles. PMID- 19681607 TI - Role of nanoparticle valency in the nondestructive magnetic-relaxation-mediated detection and magnetic isolation of cells in complex media. AB - Nanoparticle-based diagnostics typically involve the conjugation of targeting ligands to the nanoparticle to create a sensitive and specific nanosensor that can bind and detect the presence of a target, such as a bacterium, cancer cell, protein, or DNA sequence. Studies that address the effect of multivalency on the binding and detection pattern of these nanosensors, particularly on magnetic relaxation nanosensors that sense the presence of a target in a dose-dependent manner by changes in the water relaxation times (DeltaT2), are scarce. Herein, we study the effect of multivalency on the detection profile of cancer cells and bacteria in complex media, such as blood and milk. In these studies, we conjugated folic acid at two different densities (low-folate and high-folate) on polyacrylic-acid-coated iron oxide nanoparticles and studied the interaction of these magnetic nanosensors with cancer cells expressing the folate receptor. Results showed that the multivalent high-folate magnetic relaxation nanosensor performed better than its low folate counterpart, achieving single cancer cell detection in blood samples within 15 min. Similar results were also observed when a high molecular weight anti-folate antibody (MW 150 kDa) was used instead of the low molecular weight folic acid ligand (MW 441.4 kDa), although better results in terms of sensitivity, dynamic range, and speed of detection were obtained when the folate ligand was used. Studies using bacteria in milk suspensions corroborated the results observed with cancer cells. Taken together, these studies demonstrate that nanoparticle multivalency plays a key role in the interaction of the nanoparticle with the cellular target and modulate the behavior and sensitivity of the assay. Furthermore, as detection with magnetic relaxation nanosensors is a nondestructive technique, magnetic isolation and further characterization of the cancer cells is possible. PMID- 19681610 TI - Astragalosides isolated from the root of astragalus radix inhibit the formation of advanced glycation end products. AB - Because advanced glycation end product (AGE) inhibitors such as pyridoxamine significantly inhibit the development of retinopathy and neuropathy in the streptozotocin-induced diabetic rat, treatment with AGE inhibitors is believed to be a potential strategy for the prevention of lifestyle-related diseases such as diabetic complications. A crude extract of Astragali Radix (AR; roots of Astragalus membranaceus ) inhibits the formation of N(epsilon) (carboxymethyl)lysine (CML) and pentosidine during the incubation of bovine serum albumin with ribose. In the present study, compounds were isolated from AR that prevented CML and pentosidine formation. Astragalosides significantly inhibited the formation of both CML and pentosidine, and astragaloside V had the strongest inhibitory effect among all if the isolated compounds. These data suggest that AR and astragalosides may be a potentially useful strategy for the prevention of clinical diabetic complications by inhibiting AGEs. PMID- 19681609 TI - Portability of the GN(R)A hairpin loop motif between RNA and DNA. AB - Hairpins are common nucleic acid secondary structures that serve many structural and functional roles. Recently, we reported that r(UNCG) and r(GNRA) hairpin families use molecular mimicry and electrostatic factors to attain exceptional thermodynamic stability with a CG closing base pair (cbp). Despite having very different overall folds, these tetraloops present the same functionalities and partial charges to the major groove edge of the CG cbp to achieve stability. Herein, we compare the r(GNRA) tetraloop family to the DNA triloop family d(GNA), which is also exceptionally stable with a CG cbp and possesses the same base pairing between the first and last positions of the loop. Nucleobase and functional group modifications were used to investigate interactions of d(GNA) loops with the cbp, which provided for comparison with similar substitutions in r(GNRA) hairpins. Interruption or deletion of loop-cbp interactions in d(GNA) was consistent with electrostatic interactions identified through nonlinear Poisson Boltzmann (NLPB) calculations, and loop stability changed in a manner consistent with similar loop-cbp interactions for d(GNA) and r(GNRA) loops. We also compared the relationship of DeltaG degrees (37) and log[Na+] for d(GNA) and r(GNRA) loops and found a decreased dependence of stability on salt for both loop families when a CG cbp was present. The similarity of the loop-cbp interactions shows portability of this loop-cbp motif across polymer type and loop size and indicates convergence on similar molecular solutions for stability in RNA and DNA. PMID- 19681611 TI - Study of chemical changes produced in virgin olive oils with different phenolic contents during an accelerated storage treatment. AB - Chemical changes produced in an extra virgin olive oil sample in the presence (EVOO) and absence (EVOOP) of its phenolic fraction during an accelerated storage treatment at 60 degrees C up to 7 weeks were studied. Modifications in phenol content, as well as changes in several quality parameters (free acidity, peroxide value, UV absorbance, fatty acid composition, oxidative stability index, and tocopherol content) were also evaluated under the same storage conditions and compared to those of the same sample deprived of phenolic compounds. When the phenolic extract of the EVOO was studied, a decrease of the antioxidants first present in the sample and an increase of the oxidized products were observed. In addition, oxidation seemed to produce the transformation of such phenolic compounds as secoiridoids and the appearance of oxidized forms of them. These latter compounds could be used as molecular markers of the lack of extra virgin olive oil freshness. PMID- 19681612 TI - Metabolic changes in different developmental stages of Vanilla planifolia pods. AB - The metabolomic analysis of developing Vanilla planifolia green pods (between 3 and 8 months after pollination) was carried out by nuclear magnetic resonance (NMR) spectroscopy and multivariate data analysis. Multivariate data analysis of the (1)H NMR spectra, such as principal component analysis (PCA) and partial least-squares-discriminant analysis (PLS-DA), showed a trend of separation of those samples based on the metabolites present in the methanol/water (1:1) extract. Older pods had a higher content of glucovanillin, vanillin, p hydroxybenzaldehyde glucoside, p-hydroxybenzaldehyde, and sucrose, while younger pods had more bis[4-(beta-D-glucopyranosyloxy)-benzyl]-2-isopropyltartrate (glucoside A), bis[4-(beta-D-glucopyranosyloxy)-benzyl]-2-(2-butyl)tartrate (glucoside B), glucose, malic acid, and homocitric acid. A liquid chromatography mass spectrometry (LC-MS) analysis targeted at phenolic compound content was also performed on the developing pods and confirmed the NMR results. Ratios of aglycones/glucosides were estimated and thus allowed for detection of more minor metabolites in the green vanilla pods. Quantification of compounds based on both LC-MS and NMR analyses showed that free vanillin can reach 24% of the total vanillin content after 8 months of development in the vanilla green pods. PMID- 19681613 TI - Effects of date ( Phoenix dactylifera L., Medjool or Hallawi Variety) consumption by healthy subjects on serum glucose and lipid levels and on serum oxidative status: a pilot study. AB - The present pilot study analyzed, for the first time, the in vivo effect of Medjool or Hallawi date consumption by healthy subjects on serum glucose, lipids, and oxidative stress. Total phenolics concentration in the Hallawi versus Medjool dates was greater by 20-31%. The major proportion of the soluble phenolics in both date varieties consisted of phenolic acids, mainly ferulic acid and coumaric acid derivatives, and also chlorogenic and caffeic acid derivatives. Unlike the Medjool dates, Hallawi dates contained a significant proportion of catechins as well. In addition, both varieties contained a quercetin derivative. Both date varieties possess antioxidative properties in vitro, but the ferric ion reducing antioxidant power of Hallawi versus Medjool dates was higher by 24%. Ten healthy subjects consumed, for a period of 4 weeks 100 g/day of either Medjool or Hallawi dates. The date consumption did not significantly affect the subjects' body mass index (BMI), their serum total cholesterol, or their cholesterol levels in the VLDL, LDL, or HDL fractions. Most important, fasting serum glucose and triacylglycerol levels were not increased after consumption of either date variety, and serum triacylglycerol levels even significantly (p < 0.05) decreased, by 8 or 15% after Medjool or Hallawi date consumption, respectively. Basal serum oxidative status was significantly (p < 0.01) decreased by 33%, as compared to the levels observed before consumption, after Hallawi (but not Medjool) date consumption. Similarly, the susceptibility of serum to AAPH-induced lipid peroxidation decreased by 12%, but only after Hallawi date consumption. In agreement with the above results, serum activity of the HDL-associated antioxidant enzyme paraoxonase 1 (PON1) significantly increased, by 8%, after Hallawi date consumption. It is concluded that date consumption (and mainly the Hallawi variety) by healthy subjects, despite their high sugar content, demonstrates beneficial effects on serum triacylglycerol and oxidative stress and does not worsen serum glucose and lipid/lipoprotein patterns, and thus can be considered an antiatherogenic nutrient . PMID- 19681614 TI - Preparation of acetochlor antibody and its application on immunoaffinity chromatography cleanup for residue determination in peanuts. AB - The preparation of an antibody against acetochlor and its application on immunoaffinity chromatography (IAC) cleanup was investigated. The hapten was synthesized by reacting acetochlor with 3-mercaptopropionic acid and then linked with a carrier protein by the carbodiimide method. After immunizing a rabbit with hapten-bovine serum albumin (BSA) conjugate, the obtained antiserum showed an IC(50) value of 0.2 ng/mL. By coupling the purified polyclonal antibody with CNBr activated Sepharose 4B, an IAC column was prepared. The dynamic column capacity was 5560 ng/mL gel. The IAC was then used to clean up a peanut sample solution, followed by high-performance liquid chromatography (HPLC) separation and ultraviolet (UV) detection. The recoveries of acetochlor from spiked peanut samples at levels of 5-100 microg/kg ranged from 94.7 to 102.5%, with a variation coefficient of 2.6-8.0%, and the limits of detection and quantification were 1.5 and 5 microg/kg, respectively. PMID- 19681615 TI - Design, synthesis, and fungicidal activity of novel analogues of pyrrolnitrin. AB - A series of novel analogues of pyrrolnitrin containing a thiophene moiety were designed and synthesized by a facile method, and their structures were characterized by (1)H nuclear magnetic resonance (NMR) and high-resolution mass spectrometry. The isomers IV-h and V-h were isolated, and their structures were identified by 2D NMR, including heteronuclear multiple-quantum coherence (HMQC), heteronuclear multiple-bond correlation (HMBC), and nuclear Overhauser effect spectrometry (NOESY) spectra. Their fungicidal activities against five fungi were evaluated, and the results indicated that some of the title compounds showed excellent fungicidal activities in vitro against Alternaria solani , Gibberella zeae , Physalospora piricola , Fusarium omysporum , and Cercospora arachidicola at the dosage of 50 microg mL(-1). Some compounds shown moderate activity at low dosage. Compound V-h could be considered as a leading structure for further design of agricultural fungicides. PMID- 19681616 TI - Unique tautomeric and recognition properties of thioketothymines? AB - The tautomeric and recognition properties of thymine, 2- and 4-thioketothymines have been studied by means of accurate ab initio methods combined with molecular dynamics simulations and free energy calculations. In contrast to previous suggestions in the literature, the replacement of carbonyl oxygens by sulfur atoms does not lead to dramatic changes in tautomeric properties of the pyrimidine derivatives neither in vacuum nor in aqueous solution. Moreover, the presence of thioketothymines induces only mild changes in DNA structure, stability and fidelity. Despite the fact that mismatching can largely stabilize minor tautomeric forms, thioketothymines are found in the canonical thioketo-form irrespective of the paired base. Our theoretical results, confirmed by new experimental studies, describe the complete tautomeric and recognition characteristics of thioketothymines and demonstrate that both 2-thioketo and 4 thioketothymine are excellent molecules to introduce special chemical properties in modified DNA. PMID- 19681617 TI - Characterization of iron-carbonyl-protected gold clusters. AB - Ligand-stabilized nanometer-sized gold particles are interesting building blocks for molecular electronics, precursors for catalysts, optical labels for biomolecules and diagnosis, and potential nontoxic carriers for therapeutics. In this work we characterize for the first time, by means of near-infrared and Raman spectroscopy and time-dependent density functional calculations, gold clusters protected with iron-carbonyl ligands, such as {Au(22)[Fe(CO)(4)](12)}(6-) shown in the figure. Surprisingly, our results show that these novel compounds bear many analogues to another, well-studied, class of gold clusters, namely those of thiolate-monolayer-protected gold clusters. Our work adds a new dimension to the rich chemistry of gold-based clusters that can be characterized as "molecular metals". PMID- 19681618 TI - Holographic enzyme inhibition assays for drug discovery. AB - Optical sensors are widely utilized in drug discovery to analyze biomolecular interactions in vitro. Aside from additional time and cost demands, other issues associated with labeled screening methods include signal interference that can arise from the label per se and/or the screened compounds. This report describes an enzyme inhibition-based holographic sensor as a potential label-free detection system, using acetylcholinesterase (acetylcholine acetylhydrolase; EC 3.1.1.7; abbreviated herein AChE) as the model enzyme. pH-responsive reflection holograms, incorporated into "smart" hydrogel films bearing ionizable monomers, were used to monitor the pH change resulting from acetic acid produced by the hydrolysis of the substrate acetylcholine. The enzyme was immobilized on the sensor by an entrapment and in situ cross-linking method; no chemical modification and/or prelabeling of the enzyme (or the substrate) was required. The fully reversible sensor exhibited good operational and storage stability, allowing relatively short assay times and repeated use of a single sensor. Apparent inhibition parameters for several drug inhibitors of the enzyme were determined. The feasibility of adapting these sensors into an array format for prospective high throughput screening, without compromising their intrinsic optical and functional properties, was also demonstrated. PMID- 19681619 TI - Single-crystalline EuF3 hollow hexagonal microdisks: synthesis and application as a background-free matrix for MALDI-TOF-MS analysis of small molecules and polyethylene glycols. AB - Single-crystalline EuF(3) hexagonal microdisks with hollow interior were fabricated to serve as a background-free matrix for analysis of small molecules and polyethylene glycols (PEGs) by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The long-lived excited state of europium ions can transfer energy to high-energy vibrations of organic molecules, which provides the potential technological application in MALDI-TOF-MS analysis of small molecules and PEGs. The efficiency of the hollow microdisks as a novel matrix of low molecular weight compounds was verified by analysis of small peptide, amino acid, organic compounds, and hydroxypropyl beta-cyclodextrin (HP beta-CD). The advantage of this matrix in comparison with alpha-cyano-4 hydroxycinnamic acid (CHCA) and 2,5-dihydroxybenzoic acid (DHB) was demonstrated by MALDI-TOF-MS analysis of an amino acid mixture and a peptide mixture. This matrix is successfully used for analysis of PEGs (PEG 2000, PEG 4000, PEG 8000, PEG 15000, and PEG 30000), suggesting a potential for monitoring reactions and for synthetic polymer quality control. The upper limit of detectable mass range was approximately 35,000 Da (PEG 30000). It is believed that this work will not only offer a new technique for high-speed analysis of small molecules and PEGs but also open a new field for applications of rare earth fluorides. PMID- 19681620 TI - Importance of using complementary process analyzers for the process monitoring, analysis, and understanding of freeze drying. AB - The aim of the present paper is to demonstrate the importance of using complementary process analyzers (PAT tools) for the process monitoring, analysis, and understanding of freeze drying. A mannitol solution was used as a model system. Raman spectroscopic, near-infrared (NIR) spectroscopic, plasma emission spectroscopic, and wireless temperature measurements (TEMPRIS) were simultaneously performed in-line and real-time during each freeze-drying experiment. The combination of these four process analyzers to monitor a freeze drying process is unique. The Raman and NIR data were analyzed using principal component analysis (PCA) and multivariate curve resolution (MCR), while the plasma emission spectroscopic and wireless temperature measurement data were analyzed using univariate data analysis. It was shown that the considered process analyzers do not only complement but also mutually confirm each other with respect to process step end points, physical phenomena occurring during freeze drying (process understanding), and product characterization (solid state). Furthermore and most important, the combined use of the process analyzers helped to identify flaws in previous studies in which these process analyzers were studied individually. Process analyzers might wrongly indicate that some process steps are fulfilled. Finally, combining the studied process analyzers also showed that more information per process analyzer can be obtained than previously described. A combination of Raman and plasma emission spectroscopy seems favorable for the monitoring of nearly all critical freeze-drying process aspects. PMID- 19681621 TI - 91Zr nuclear magnetic resonance spectroscopy of solid zirconium halides at high magnetic field. AB - (91)Zr solid-state NMR spectra of zirconium halides and several fluorozirconates have been obtained at high magnetic fields up to 30 T using both the Hahn-Echo and the Quadrupolar Carr-Purcell-Meiboom-Gill sequences combined with the broadband Variable Offset Cumulative Spectrum technique. For the zirconium halides, the (91)Zr isotropic chemical shift covers a range of about 2000 ppm and shows a good correlation with Pauling's electronegativity and ionic potential of the halogen. For the fluorozirconate samples, in which the Zr atoms exhibit various coordination polyhedra, increasing the Zr coordination number and the mean Zr-F bond length leads to an increased isotropic shielding. In the studied compounds the (91)Zr quadrupolar coupling constants (C(Q)'s) range from 10.6 to 44.7 MHz. For 6-fold coordinated Zr sites, a correlation between C(Q) and the shear strain of the octahedron is observed, and we investigate the relationship between the C(Q) and the distortion of the polyhedron for 8-fold coordinated Zr sites using different distortion criteria. PMID- 19681622 TI - Accurate double many-body expansion potential energy surface for N3((4)A'') from correlation scaled ab initio energies with extrapolation to the complete basis set limit. AB - A new global potential energy surface is reported for the (4)A'' ground electronic state of the N(3) system from double many-body expansion theory and an extensive set of accurate ab initio energies extrapolated to the complete basis set limit. It shows three equivalent metastable potential wells for C(2v) geometries that are separated from the three N((4)S) + N(2) asymptotes by energy barriers as predicted from previous ab initio work. The potential well and barrier height now predicted lie 42.9 and 45.9 kcal mol(-1) above the atom-diatom dissociation limit, respectively, being about 1 kcal mol(-1) lower than previous theoretical estimates. The ab initio calculations here reported predict also a (4)B(1)/(4)A(2) conical intersection and reveal a new minimum with D(3h) symmetry that lies 147 kcal mol(-1) above the atom-diatom asymptote. All major topographical features of the potential energy surface are accurately described by the DMBE function, including the weakly bound van der Waals minima at large atom-diatom separations. PMID- 19681624 TI - Inelastic scattering of the NCO(X2Pi) radical with the He atom on an ab initio potential energy surface. AB - The (2)A' and (2)A'' adiabatic potential energy surfaces for the He-NCO(X(2)Pi) van der Waals system are obtained by the partially spin-restricted coupled cluster method with single, double, and noniterated triple excitations (RCCSD(T)). The ab initio potentials are fit to analytical expressions, and scattering and bound state calculations are performed for a rigid NCO((2)Pi) radical. Rotational constants of the complex are reported. The scattering calculations of integral and differential cross sections are performed using both the fully quantum close-coupling (CC) and coupled-states (CS) methods. The collision energies have the values taken from the experiment of Macdonald and Liu (J. Chem. Phys. 1992, 97, 978). The excellent agreement between theoretical and experimental scattering results attests good quality of the ab initio potential. PMID- 19681623 TI - Application of multipolar charge models and molecular dynamics simulations to study stark shifts in inhomogeneous electric fields. AB - Atomic multipole moments are used to investigate vibrational frequency shifts of CO and H(2) in uniform and inhomogeneous electric fields using ab initio calculations and Molecular Dynamics (MD) simulations. The importance of using atomic multipole moments that can accurately represent both molecular electrostatics and the vibrational response of the molecule to changes in the local electric field is highlighted. The vibrational response of CO to applied uniform and inhomogeneous electric fields is examined using Density Functional Theory calculations for a range of test fields, and the results are used to assess the performance of different atomic multipole models. In uniform fields, the calculated Stark tuning rates of Deltamu = 0.52 cm(-1)/(MV/cm) (DFT), Deltamu = 0.55 cm(-1)/(MV/cm) (fluctuating three-point charge model), and Deltamu = 0.64 cm(-1)/(MV/cm) (Multipole model up to octupole), compare favorably with the experimentally measured value of 0.67 cm(-1)/(MV/cm). For H(2), which has no permanent dipole moment, CCSD(T) calculations demonstrate the importance of bond weakening effects in force fields in response to the applied inhomogeneous electric field. Finally, CO in hexagonal ice is considered as a test system to highlight the performance of selected multipolar models in MD simulations. The approach discussed here can be applied to calibrate a range of multipolar charge models for diatomic probes, with applications to interpret Stark spectroscopy measurements in protein active sites. PMID- 19681625 TI - Organic single-crystal surface-induced polymerization of conducting polypyrroles. AB - Polypyrrole hexagonal microplates (PHMs) (50-100 microm long, 10-20 microm wide, and 0.8-1.2 microm thick) with a quasicrystalline structure and high electrical conductivity (up to 400 S/cm) are simply fabricated using single crystals of 4 sulfobenzoic acid monopotassium salt (KSBA) in aqueous medium. Moreover, the fabrication process described here differs strikingly from traditional methods, such as template-free, soft template, and hard template methods. Synthetic time resolved polypyrrole (PPy) morphology dynamics reveals that the fabrication process of PHMs composed of PPy nanostructures combines a shape-copying process for forming a PPy preform that imitates the shape of a KSBA single crystal and the self-assembly process of PPys on the preform. The PHMs exhibit the improved pi-stacking and bipolaron structure. The strong pi-stacks among PPy rings of bipolaron structures lead to a high quasicrystalline structural order and the metallic conduction. Other single organic crystals that can act as dopants could also be grown using this approach, which will also enable the fabrication of complex micro/nanostructures on organic single crystals. PMID- 19681626 TI - Role of linker groups between hydrophilic and hydrophobic moieties of cationic surfactants on oligonucleotide-surfactant interactions. AB - The interaction between DNA and amino-acid-based surfactants with different linker groups was investigated by gel electrophoresis, ethidium bromide exclusion assays, circular dichroism, and melting temperature determinations. The studies showed that the strength of the interaction between the oligonucleotides and the surfactants is highly dependent on the linker of the surfactant. For ester surfactants, no significant interaction was observed for surfactant-to-DNA charge ratios up to 12. On the other hand, amide surfactants were shown to interact strongly with the oligonucleotides; these surfactants could displace up to 75% of the ethidium bromide molecules bound to the DNA and induced significant changes in the circular dichroism spectra. When comparing the headgroups of the surfactants, it was observed that surfactants with more hydrophobic headgroups (proline vs alanine) interacted more strongly with the DNA, in good agreement with previous studies. PMID- 19681627 TI - Visible-light-driven superhydrophilicity by interfacial charge transfer between metal ions and metal oxide nanostructures. AB - Single-crystalline rutile nanorods were synthesized by a facile acid treatment on titanate nanotubes. These rutile nanorods could be highly dispersed in water to form a stable colloidal solution. Cu(2+) ions were grafted onto these rutile nanorods, and the Cu(2+)-grafted nanorods could absorb visible light by the interfacial charge transfer between the valence band of rutile TiO(2) and surface modified Cu(2+) ions. Transparent thin films of Cu(2+)-grafted nanorods were coated on substrates by a facile spin-coating method. These films exhibited superhydrophilic conversion under visible light irradiation. PMID- 19681628 TI - Gemini surfactant electrospun membranes. AB - Our research demonstrates electrospun nonwoven fibrous scaffolds from a low molar mass gemini ammonium surfactant, N,N'-didodecyl-N,N,N',N'-tetramethyl-N,N' ethanediyldiammonium dibromide (12-2-12). Cryogenic transmission electron microscopy (cryo-TEM) and solution rheological experiments revealed micellar morphological transitions of 12-2-12 in water and water:methanol (1:1 vol). The microstructure of 12-2-12 in water transitioned from entangled, cylindrical, threadlike micelles to branched threadlike micelles, and a viscoelastic, entangled, highly branched network of threadlike micelles with increasing concentration finally formed. In sharp contrast, the solution behavior of 12-2-12 in water:methanol produced a drastically different micellar microstructure compared to that in water, and the morphology transitioned from partitioned, globular micelles to overlapped micelles at an overlap concentration (C*) of 11 wt %. Electrospinning 12-2-12 from water did not produce fibers at any concentration; however, electrospinning 12-2-12 in water:methanol at concentrations greater than 2C* produced hydrophilic continuous fibers with diameters ranging from 0.9 to 7 microm. High surface area scaffolds with hydrophilic surfaces offer potential as charged controlled-release membranes, tissue engineering scaffolds, and coatings for biologically compatible devices. PMID- 19681629 TI - Can the C(5)H(5) + C(5)H(5) --> C(10)H(10) --> C(10)H(9) + H/C(10)H(8) + H(2) reaction produce naphthalene? An Ab initio/RRKM study. AB - Ab initio and density functional calculations using a variety of theoretical methods (CASSCF, B3LYP, CASPT2, CCSD(T), and G3(MP2,CC)) have been carried out to unravel the mechanism of unimolecular isomerization and dissociation of 9,10 dihydrofulvalene C(10)H(10) (S0) formed by barrierless recombination of two cyclopentadienyl radicals. Different reaction pathways on the C(10)H(10) potential energy surface (PES) are found to lead to the production of 9-H fulvalenyl radical + H, 9-H-naphthyl radical (a naphthalene precursor) + H, and naphthalene + H(2). RRKM calculations of thermal rate constants and product branching ratios at the high pressure limit show that at temperatures relevant to combustion the 9-H-fulvalenyl radical formed by a direct H loss from S0 with endothermicity of 76.3 kcal/mol is expected to be the dominant reaction product. The naphthalene precursor 9,10-dihydronaphthalene (D3) can be produced from the initial S0 adduct by a multistep diradical mechanism involving the formation of a metastable tricyclic diradical intermediate, followed by its three-step opening to a 10-member ring structure, which then undergoes ring contraction producing the naphthalene core structure in D3, with the highest barrier on this pathway being 70.3 kcal/mol. D3 can lose molecular hydrogen producing naphthalene via a barrier of 77.7 kcal/mol relative to the initial adduct. Another possibility is a hydrogen atom elimination in D3 giving rise to the 9-H-naphthyl radical without exit barrier and with overall endothermicity of 59.2 kcal/mol. The pathway to 9-H naphthyl appears to be preferable as compared to the direct route to 9-H fulvalenyl at temperatures below 600 K, but the rate constants at these temperatures are too slow for the reaction to be significant. The naphthalene + H(2) channel is not viable at any temperature. The following reaction sequence is suggested for kinetic models to account for the recombination of two cyclopentadienyl radicals: c-C5H5+c-C5H5-->9,10-dihydrofulvalene-->9-H fulvalenyl+H(C10H10PES), 9-H-fulvalenyl-->naphthalene+H/fulvalene+H(C10H9PES). We conclude that naphthalene can be produced from the recombination of two cyclopentadienyl radicals and is expected to be a favorable product of this reaction sequence at T < 1000 K, but this molecule would be formed through isomerizations and H atom loss on the C(10)H(9) PES (after the initial H elimination from C(10)H(10) S0) and not in conjunction with molecular hydrogen. The alternative product, fulvalene, can potentially contribute to the growth of cyclopentafused polycyclic aromatic hydrocarbons. PMID- 19681630 TI - Conformational study of taurine in the gas phase. AB - The conformational preferences of the amino sulfonic acid taurine (NH(2)-CH(2) CH(2)-SO(3)H) have been investigated in the gas phase by laser ablation molecular beam Fourier transform microwave spectroscopy (LA-MB-FTMW) in the 6-14 GHz frequency range. One conformer has been observed, and its rotational, centrifugal distortion, and hyperfine quadrupole coupling constants have been determined from the analysis of its rotational spectrum. Comparison of the experimental constants with those calculated theoretically identifies the detected conformer unambiguously. The observed conformer of taurine is stabilized by an intramolecular hydrogen bond O-H...N between the hydrogen of the sulfonic acid group and the nitrogen atom of the amino group. PMID- 19681631 TI - Structural evolution, sequential oxidation, and chemical bonding in tritantalum oxide clusters: Ta(3)O(n)(-) and Ta(3)O(n) (n = 1-8). AB - We report a combined photoelectron spectroscopy (PES) and density functional theory (DFT) study on a series of tritantalum oxide clusters, Ta(3)O(n)(-). Well resolved PES spectra are obtained for Ta(3)O(n)(-) (n = 1-8) at several detachment photon energies, yielding electronic structure information which is used for comparison with the DFT calculations. A trend of sequential oxidation is observed as a function of O content until Ta(3)O(8)(-), which is a stoichiometric cluster. Extensive DFT calculations are performed in search of the lowest energy structures for both the anions and neutrals. The first three O atoms are shown to successively occupy the bridging sites in the Ta(3) triangle. The next three O atoms each occupy a terminal site, with the seventh and eighth O atoms forming a double-bridge and a double-terminal, respectively. The Ta(3)O(7)(-) anion is found to possess a localized electron pair on a single Ta center, making it an interesting molecular model for Ta(3+) surface sites. Molecular orbital analyses are performed to elucidate the chemical bonding in the Ta(3)O(n)(-) clusters. PMID- 19681632 TI - On the thermal conductivity of gold nanoparticle colloids. AB - Nanofluids (colloidal suspensions of nanoparticles) have been reported to display significantly enhanced thermal conductivities relative to those of conventional heat transfer fluids, also at low concentrations well below 1% per volume (Putnam, S. A., et at. J. Appl. Phys. 2006, 99, 084308; Liu, M.-S. L., et al. Int. J. Heat Mass Transfer. 2006, 49; Patel, H. E., et al. Appl. Phys. Lett. 2003, 83, 2931-2933). The purpose of this paper is to evaluate the effect of the particle size, concentration, stabilization method and particle clustering on the thermal conductivity of gold nanofluids. We synthesized spherical gold nanoparticles of different size (from 2 to 45 nm) and prepared stable gold colloids in the range of volume fraction of 0.00025-1%. The colloids were inspected by UV-visible spectroscopy, transmission electron microscope (TEM) and dynamic light scattering (DLS). The thermal conductivity has been measured by the transient hot-wire method (THW) and the steady state parallel plate method (GAP method). Despite a significant search in parameter space no significant anomalous enhancement of thermal conductivity was observed. The highest enhancement in thermal conductivity is 1.4% for 40 nm sized gold particles stabilized by EGMUDE (triethyleneglycolmono-11-mercaptoundecylether) and suspended in water with a particle-concentration of 0.11 vol%. PMID- 19681633 TI - Directed assembly of SWNTs by electrostatic interactions and its application for making network transistors. AB - Single-wall carbon nanotubes (SWNTs) are promising building blocks for the fabrication of nanoelectronic devices. However, achieving control over their assembly on substrates has been challenging and is still a bottleneck to their utilization. Herein, we present a general method for directing the chemical assembly of SWNTs on substrates through electrostatic interactions. By covalently functionalizing both the nanotube sidewalls and the SiO(2) substrate with charged groups, dense networks of SWNTs were produced. The method is selective and highly efficient to process network field-effect transistors. PMID- 19681634 TI - Effects of pressure and temperature on the self-assembled fully hydrated nanostructures of monoolein-oil systems. AB - Synchrotron small-angle X-ray scattering (SAXS) was applied for studying the effects of hydrostatic pressure and temperature on the structural behavior of fully hydrated tetradecane (TC)-loaded monoolein (MO) systems. Our main attention focused on investigating the impact of isobaric and isothermal changes on the stability of the inverted type discontinuous Fd3m cubic phase as compared to the inverted type hexagonal (H(2)) liquid crystalline phase. The present results show that compressing the TC-loaded Fd3m phase under isothermal conditions induces a significant increase of its lattice parameter: it approximately increases by 1 A per 75 bar. Further, the Fd3m phase is more pressure-sensitive as compared to the Pn3m and the H(2) phases. At ambient temperatures, we observed the following structural transitions as pressure increases: Fd3m --> H(2) --> Pn3m. Our findings under isobaric conditions reveal more complicated structural transitions. At high pressures, we recorded the interesting temperature-induced structural transition of (Pn3m + L(alpha)) --> (Pn3m + L(alpha) + H(2)) --> (L(alpha) + H(2)) --> H(2) --> Fd3m --> traces of Fd3m coexisting with L(2). At high pressures and low temperatures, the TC molecules partially crystallize as indicated by the appearance of an additional diffraction peak at q = 3.46 nm(-1). This crystallite disappears at high temperatures and also as the system gets decompressed. The appearance of the Pn3m and the L(alpha) phases during compressing the fully hydrated MO/TC samples at high pressures and low temperatures is generally related to a growing hydrocarbon chain condensation, which leads to membrane leaflets with less negative interfacial curvatures (decreasing the spontaneous curvatures |H(0)|). Both the effects of pressure and temperature are discussed in detail for all nonlamellar phases on the basis of molecular shape and packing concepts. PMID- 19681635 TI - Density enhancement of aligned single-walled carbon nanotube thin films on quartz substrates by sulfur-assisted synthesis. AB - The density of the aligned single-walled carbon nanotubes (SWNTs) grown on quartz substrates is an important factor for the performance of fabricated electronic devices. It was discovered that the addition of a sulfur-containing compound (thiophene) to the reaction mixture improved the density of SWNTs by a factor of 2 or more, from approximately 2-4 SWNTs/microm to 6-8 SWNTs/microm under similar growth conditions. It was also observed that along with the increase in nanotube density, the cleanness of the samples improved as well. These effects were demonstrated over a large range of growth conditions, indicating that the addition sulfur makes the growth processes more favorable for the nucleation and growth of aligned SWNTs. PMID- 19681636 TI - Fullerenol-cytotoxic conjugates for cancer chemotherapy. AB - In the present study, we report the novel application of polyhydroxylated fullerenes (fullerenols) in cancer drug delivery. The facile synthetic procedure for generating multiple hydroxyl groups on the fullerene cage offers scope for high drug loading in addition to conferring hydrophilicity. Doxorubicin, a first line cancer chemotherapeutic, was conjugated to fullerenols through a carbamate linker, achieving ultrahigh loading efficiency. The drug-fullerenol conjugate was found to be relatively stable in phosphate buffer saline but temporally released the active drug when incubated with tumor cell lysate. The fullerenol-doxorubicin conjugate suppressed the proliferation of cancer cell-lines in vitro through a G2 M cell cycle block, resulting in apoptosis. Furthermore, in an in vivo murine tumor model, fullerenol-doxorubicin exhibited comparable antitumor efficacy as free drug without the systemic toxicity of free doxorubicin. Additionally, we demonstrate that the fullerenol platform can be extended to other chemotherapeutic agents, such as the slightly water-soluble cisplatin, and can emerge as a new paradigm in the management of cancer. PMID- 19681637 TI - Iron-catalyzed selective oxidation of N-methyl amines: highly efficient synthesis of methylene-bridged bis-1,3-dicarbonyl compounds. AB - Methylene-bridged bis-1,3-dicarbonyl derivatives were synthesized efficiently by iron-catalyzed oxidative reactions of 1,3-dicarbonyl compounds and N,N dimethylaniline. Bipyrazoles and substituted 1,4-dihydropyridine were obtained by the reactions of bis-1,3-dicarbonyl compounds with hydrazines and ammonium acetate, respectively. PMID- 19681638 TI - Self-assembly of lithographically patterned nanoparticles. AB - The construction of three-dimensional (3D) objects, with any desired surface patterns, is both critical to and easily achieved in macroscale science and engineering. However, on the nanoscale, 3D fabrication is limited to particles with only very limited surface patterning. Here, we demonstrate a self-assembly strategy that harnesses the strengths of well-established 2D nanoscale patterning techniques and additionally enables the construction of stable 3D polyhedral nanoparticles. As a proof of the concept, we self-assembled cubic particles with sizes as small as 100 nm and with specific and lithographically defined surface patterns. PMID- 19681639 TI - Building blocks for molecule-based magnets: radical anions and dianions of substituted 3,6-dimethylenecyclohexane-1,2,4,5-tetrones as paramagnetic bridging ligands. AB - We have prepared four tetraaryl derivatives of 3,6-dimethylene-1,2,4,5 tetraoxocyclohexane (aryl = Ph; 4-MeOPh; 4-Me(2)NPh; and 3,5-(t-Bu)(2)-4-MeOPh) with guidance from an earlier reported ab initio analysis (Misiolek, A. W.; Jackson, J. E. J. Am. Chem. Soc. 2001, 123, 4774-4780). These electron acceptors may be chemically or electrochemically reduced to the mono- and dianions desired as building blocks for the assembly of molecule-based magnets. Cyclic voltammetry shows that the potential of the first reduction wave depends on the electron donor ability of the aryl ring substituents, ranging from -0.28 V for the tetraphenyl derivative to -0.78 V for the p-dimethylamino substituted analogue (vs ferrocene/ferrocinium(+) at 0.46 V). Spin density distributions in the semiquinone moieties were elucidated by electron paramagnetic resonance (EPR) and electron-nuclear double resonance (ENDOR) observations of hyperfine couplings to internal (1)H sites and bound alkali metal cations. X-ray diffraction studies of the sodium and potassium salts of the octa-t-butyltetramethoxy derivative reveal the structure of the monoanion and its tendency to self-assemble with metal cations into one-dimensional chains in the solid state. Within the chains the anions display the expected bridging and chelating mode of coordination; SQUID magnetometry revealed weak intermolecular spin-spin couplings of 2J = -0.2 and approximately 0 K for the sodium and potassium salts, respectively. NIR transitions in the electronic spectra of the monoanions in solution are consistent with the expected low energy gap between frontier orbitals and its tunability by substituent variations. EPR studies of the free dianions and monoradical analogues indicate diradical localization into separate triphenylmethyl-like monoradicals via twisting of the diarylmethylene termini. PMID- 19681640 TI - 4-Amino-1,8-naphthalimide-based Troger's bases as high affinity DNA targeting fluorescent supramolecular scaffolds. AB - The synthesis and photophysical and biological investigation of fluorescent 1,8 naphthalimide conjugated Troger's bases 1-3 are described. These structures bind strongly to DNA in competitive media at pH 7.4, with concomitant modulation in their fluorescence emission. These structures also undergo rapid cellular uptake, being localized within the nucleus within a few hours, and are cytotoxic against HL60 and (chronic myeloid leukemia) K562 cell lines. PMID- 19681641 TI - Controlling the electronic properties of silicon nanowires with functional molecular groups. AB - Attachment of a simple polar molecule, terpyridine, to the surfaces of Si nanowires with diameters ranging from 7-100 nm was investigated with two terminal conductivity measurements, a unique single nanowire photoelectron spectroscopy, and theoretical analysis. Our experiments reveal the details of molecule-nanowire bonding and charge transfer, diameter dependent Fermi level shifts, and acid attachment leading to conductivity in the cylindrical molecular nanolayer surrounding the nanowire. PMID- 19681643 TI - Hydrophilic anthropogenic markers for quantification of wastewater contamination in ground- and surface waters. AB - Hydrophilic, persistent markers are useful to detect, locate, and quantify contamination of natural waters with domestic wastewater. The present study focused on occurrence and fate of seven marker candidates including carbamazepine (CBZ), 10,11-dihydro-10,11-dihydroxycarbamazepine (DiOH-CBZ), primidone (PMD), crotamiton (CTMT), N-acetyl-4-aminoantipyrine (AAA), N-formyl-4-aminoantipyrine (FAA), and benzotriazole (BTri) in wastewater treatment plants (WWTPs), lakes, and groundwater. In WWTPs, concentrations from 0.14 microg/L to several micrograms per liter were observed for all substances, except CTMT, which was detected at lower concentrations. Loads determined in untreated and treated wastewater indicated that removal of the potential markers in WWTPs is negligible; only BTri was partly eliminated (average 33%). In lakes, five compounds, CBZ, DiOH-CBZ, FAA, AAA, and BTri, were consistently detected in concentrations of 2 to 70 ng/L, 3 to 150 ng/L, less than the limit of quantification to 30 ng/L, 2 to 80 ng/L, and 11 to 920 ng/L, respectively. Mean per capita loads in the outflows of the lakes suggested possible dissipation in surface waters, especially of AAA and FAA. Nevertheless, concentrations of CBZ, DiOH-CBZ, and BTri correlated with the actual anthropogenic burden of the lakes by domestic wastewater, indicating that these compounds are suitable for quantification of wastewater contamination in lakes. Marker candidates were also detected in a number of groundwater samples. Carbamazepine concentrations up to 42 ng/L were observed in aquifers with significant infiltration of river water, receiving considerable wastewater discharges from WWTPs. Concentration ratios between compounds indicated some elimination of BTri and DiOH-CBZ during subsurface passage or in groundwater, while CBZ and PMD appeared to be more stable and thus are promising wastewater markers for groundwater. The wastewater burden in groundwater, estimated with the markers CBZ and PMD, reached up to 6%. PMID- 19681644 TI - Environmental safety aspects of personal care products--a European perspective. PMID- 19681642 TI - Structural basis for differential binding of the interleukin-8 monomer and dimer to the CXCR1 N-domain: role of coupled interactions and dynamics. AB - Interleukin-8 (IL-8 or CXCL8) plays a critical role in orchestrating the immune response by binding and activating the receptor CXCR1 that belongs to the GPCR class. IL-8 exists as both monomers and dimers, and both bind CXCR1 but with differential affinities. It is well established that the monomer is the high affinity ligand and that the interactions between the ligand N-loop and receptor N-domain play a critical role in determining binding affinity. In order to characterize the structural basis of differential binding of the IL-8 monomer and dimer to the CXCR1 N-domain, we analyzed binding-induced NMR chemical shift and peak intensity changes and show that they are exquisitely sensitive and can provide detailed insights into the binding process. We used three IL-8 variants, a designed monomer, a trapped disulfide-linked dimer, and WT at dimeric concentrations. NMR data for the monomer show that nonsequential residues that span the entire N-loop are involved in the binding process and that the binding is mediated by a network of extensive direct and indirect coupled interactions. Interestingly, in the case of WT, binding induces dissociation of the dimer receptor complex to the monomer-receptor complex, and in the case of the trapped dimer, binding results in increased global conformational flexibility. Increased dynamics is evidence of unfavorable interactions, indicating that binding of the WT dimer triggers conformational changes that disrupt dimer-interface interactions, resulting in its dissociation. These results together provide evidence that binding is not a localized event but results in extensive coupled interactions within the monomer and across the dimer interface and that these interactions play a fundamental role in determining binding affinity. PMID- 19681645 TI - Chemicals from the practice of healthcare: challenges and unknowns posed by residues in the environment. PMID- 19681646 TI - Prevalence and the associated lesions of Cheilospirura (Acuaria) hamulosa in the indigenous chicken of Kashmir Valley, India. AB - The present investigation reports the results from a 2-yr study conducted on the prevalence and associated lesions of the nematode Cheilospirura (Acuaria) hamulosa (Nematoda, Acuarioidea) in a sample size of 478 indigenous fowl collected from different localities of Kashmir Valley, India. The investigation revealed the annual prevalence for the nematode for the first and second yr of study to be 3.4% (8/233) and 3.7% (9/245), respectively, with an overall prevalence of 3.5% (17/478). Parasitized birds did not present with clinical signs. Despite the low parasite burdens, the lesions induced by C. hamulosa were severe. Histological examination of the infected gizzard revealed discrete and coalescing nodular and cystic lesions in the mucosa and musculature which contained sections of the parasite. Cellular reaction in the lesions was characterized by a large number of lymphocytes, monocytes, plasma cells, heterophils, and, in some of the sections, severe eosinophilic reaction. The mucosa and submucosa showed markedly thickened and diffuse mononuclear infiltration as well as reactive lymphoid nodules. PMID- 19681647 TI - Histomonas meleagridis (Protozoa: Trichomonadidae): analysis of growth requirements in vitro. AB - Histomonads grew rapidly in Dwyer's medium, consisting of medium 199, chick embryo extract, serum, and rice powder, reaching a population size of about 5 x 10(5) in 3-4 days, followed by a rapid decline. Substitution of other cell culture media (L-15, MEM, or RPMI) for M199 was also satisfactory, except for Waymouth's medium, which produced a lower and later peak of growth. Omission of serum or rice rendered media unsuitable for growth. Bacteriological culture media did not support growth of histomonads. Media that included glucose were unsuitable because the pH of the cultures dropped to about 4. The effect of glucose on pH was due to the action of bacteria. Oxygen inhibited growth of histomonads. There was no growth when culture tubes were not capped tightly, regardless of the medium used. Histomonads grew well with rice flour, cornstarch, oat flour, rye flour, and buckwheat flour. Barley and blue corn meal were less satisfactory. It appeared that the requirements for growth of the lumen phase Histomonas meleagridis included a suitable physiological saline, serum (of any source), and a starch source (grain flour). Anaerobic conditions and a pH near neutral were best. Histomonads separated into pure cultures by flow cytometry would not grow without the inclusion of an unspecified species of bacteria. PMID- 19681648 TI - Influence of host biological features on macroparasites of the two European anglerfish species, Lophius piscatorius and Lophius budegassa, off north and northwest Spain. AB - Two anglerfish species, Lophius piscatorius and Lophius budegassa, caught off the north and northwest coast of Spain in 2007, were examined for parasite infections. Four parasite taxa were chosen for study, i.e., Anisakis larvae Type I (sensu Berland, 1961), Hysterothylacium rigidum (Nematoda), Spraguea lophii (Microsporidia), and Chondracanthus lophii (Copepoda). Prevalence, intensity, and abundance were determined and related to host size and sex. The abundance of Anisakis larvae Type I significantly increased with size in both anglerfish species, and C. lophii was also more abundant in larger L. piscatorius specimens. The abundance of H. rigidum and the prevalence of S. lophii were unrelated to host size. The infestation with the parasites studied was not affected by host sex. PMID- 19681649 TI - Identification of Taenia sp. in a mummy from a Christian Necropolis in El-Deir, Oasis of Kharga, ancient Egypt. AB - For the first time, a palaeoparasitological study was performed on 12 mummies from a Christian cemetery excavated in El-Deir, Oasis of Kharga, Egypt. The analysis revealed the presence of a tapeworm, probably Taenia sp., in a single individual. The presence of just the presumed taeniid egg is surprising and raises the question of the relationship between residents of Egyptian oases and those residing in the Nile Valley. The result suggests information on the health status of the ancient oasis population and re-enforces a hypothesis regarding possible social stratification of the inhabitants. The work must be continued if we are to acquire additional knowledge dealing with life in ancient Egyptian oases. PMID- 19681650 TI - Swine influenza (H1N1) pandemic: developing countries' perspective. AB - Since the first case of the current pandemic (H1N1) 2009 virus reported to WHO on 24 April 2009 on the American continent, the virus has spread in 160 countries and territories. By mid-2009, there were 135,000 cases and 816 deaths recorded. Pandemic preparedness is not advanced in most developing countries. Effective and essential measures include heightened surveillance, early detection and appropriate medical care. The use of local resources and capacity building with the assistance of developed nations will reduce the impact of this pandemic in the developing world. PMID- 19681652 TI - Ca2+ oscillations and its transporters in mesenchymal stem cells. AB - Intracellular free Ca2+ is one of important biological signals regulating a number of cell functions. It has been discussed widely and extensively in several cell types during the past two decades. Attention has been paid to the Ca2+ transportation in mesenchymal stem cells in recent years as mesenchymal stem cells have gained considerable interest due to their potential for cell replacement therapy and tissue engineering. In this paper, roles of intracellular Ca2+ oscillations and its transporters in mesenchymal stem cells have been reviewed. PMID- 19681651 TI - Remifentanil protects myocardium through activation of anti-apoptotic pathways of survival in ischemia-reperfused rat heart. AB - Remifentanil is a commonly used opioid in anesthesia with cardioprotective effect in ischemia-reperfused (I/R) heart. We evaluated the influence of remifentanil on myocardial infarct size and expressions of proteins involved in apoptosis in I/R rat heart following various time protocols of remifentanil administration. Artificially ventilated anesthetized Sprague-Dawley rats were subjected to a 30 min of left anterior descending coronary artery occlusion followed by 2 h of reperfusion. Rats were randomly assigned to one of five groups; Sham, I/R only, remifentanil preconditioning, postconditioning and continuous infusion group. Myocardial infarct size, the phosphorylation of ERK1/2, Bcl2, Bax and cytochrome c and the expression of genes influencing Ca2+ homeostasis were assessed. In remifentanil-administered rat hearts, regardless of the timing and duration of administration, infarct size was consistently reduced compared to I/R only rats. Remifentanil improved expression of ERK1/2 and anti-apoptotic protein Bcl2, and expression of sarcoplasmic reticulum genes which were significantly reduced in the I/R rats only. Remifentanil reduced expression of pro-apoptotic protein, Bax and cytochrome c. These suggested that remifentanil produced cardioprotective effect by preserving the expression of proteins involved in anti-apoptotic pathways, and the expression of sarcoplasmic reticulum genes in I/R rat heart, regardless of the timing of administration. PMID- 19681653 TI - [3H]SCH 23390 binding in various brain regions of C57BL/6J mice with repeated experience of victory or social defeat in agonistic interactions. AB - The binding of [3H]SCH 23390 has been studied in various brain regions of male mice with the experience of repeated victory (winners) or defeat (losers) gained over 10 (T10) and 20 (T20) days of daily agonistic confrontations. In the frontal cortex, B(max) of [3H]SCH 23390 binding sites was found to be increased in T10 losers and decreased in T20 losers when compared to the control mice. In the striatum, T10 and T20 winners had reduced values of [3H]SCH 23390 binding sites than the ones in the control mice. The K(d) was increased in the frontal cortex of T10 losers and T10 winners as well as in the amygdala of T20 losers. Reduced K(d) values were found in the striatum of all experimental groups as well as in the amygdala of T20 winners. Thus, both specific changes relating to social behavior patterns and non-specific ones in [3H]SCH 23390 binding were found in the brain regions of mice after 10 and 20 days of intermale confrontations. PMID- 19681654 TI - Hepcidin expression in adipose tissue increases during cardiac surgery. AB - Hepcidin, a key regulator of iron metabolism, plays a crucial role in the pathogenesis of anemia of chronic disease. Although it is produced mainly in the liver, its recently described expression in adipose tissue has been shown to be enhanced in massive obesity due to chronic low-grade inflammation. Our objective was to study the changes in hepcidin expression in adipose tissue during acute phase reaction. We measured hepcidin mRNA expression from isolated subcutaneous and epicardial adipose tissue at the beginning and at the end of the surgery. The expression of mRNAs for hepcidin and other iron-related genes (transferrin receptor 1, divalent metal transporter 1, ferritin, ferroportin) were measured by real-time RT-PCR. Hepcidin expression significantly increased at the end of the surgery in subcutaneous but not in epicardial adipose tissue. Apart from the increased levels of cytokines, the parameters of iron metabolism showed typical inflammation-induced changes. We suggest that acute inflammatory changes could affect the regulation of hepcidin expression in subcutaneous adipose tissue and thus possibly contribute to inflammation-induced systemic changes of iron metabolism. PMID- 19681655 TI - Plasma concentrations of fibroblast growth factors 21 and 19 in patients with Cushing's syndrome. AB - The objective of this study was to measure plasma fibroblast growth factor 21 and 19 (FGF21 and FGF19) levels in patients with Cushing's syndrome (CS) and to compare it with those of lean control subjects (C) and patients with obesity (OB). Fourteen untreated patients with CS, 19 patients with OB and 36 controls were included in the study. Plasma FGF21 and FGF19 levels were measured by ELISA kits, other hormonal and biochemical parameters were measured by standard laboratory methods. Plasma FGF19 did not significantly differ among the studied groups. Plasma FGF21 levels were significantly higher in both CS and OB groups relative to C group but they did not differ between CS and OB groups. In a combined population of all three groups FGF21 levels positively correlated with BMI, waist circumference and percentage of total and truncal fat mass. Less prominent inverse relationship with these parameters was found for FGF19. Neither FGF21 nor FGF19 were significantly related to cortisol concentrations. Increased FGF21 concentrations in both patients with CS and OB relative to lean subjects suggest that excessive body fat and/or related metabolic abnormalities rather than direct effects of cortisol are responsible. In contrast neither obesity nor hypercortisolism significantly affected FGF19 concentrations. PMID- 19681656 TI - Long-term efficacy and safety of conversion to tacrolimus in heart transplant recipients with ongoing or recurrent acute cellular rejection. AB - Despite the widespread use of potent immunosuppressive drugs, such as cyclosporin A and mycophenolate mofetil, ongoing and recurrent cellular rejection remain a common problem after heart transplantation. We aimed to describe the long-term effects of conversion from cyclosporine A to tacrolimus in patients with ongoing and recurrent cellular rejection. This was a single-centre retrospective analysis of 17 heart transplant recipients who were switched from cyclosporine A to tacrolimus due to ongoing (5 patients) or recurrent cellular rejection (12 patients). We studied long-term efficacy and safety of this approach. 167 endomyocardial biopsies were performed during a mean follow-up of 69.1 +/- 12.7 months. Thirteen biopsies (7.8%) in eight patients (47%) revealed higher grades of acute cellular rejection (Banff 2). However, they were not hemodynamically significant and did not require intravenous antirejection therapy. The mean rejection score was reduced significantly. Conversion to tacrolimus was tolerated in 82% pts without any significant side effects during a long-term follow-up. In conclusion, the conversion to tacrolimus in heart transplant recipients with ongoing or recurrent acute cellular rejection was safe and effective also during a long-term follow-up. PMID- 19681657 TI - May alcohol-induced increase of HDL be considered as atheroprotective? AB - It is well known that the consumption of moderate doses of alcohol leads to the increase of HDL-cholesterol (HDL-C). Atheroprotectivity of HDL particles is based primarily on their role in reverse cholesterol transport (RCT). In the study with a cross-over design 13 male volunteers were studied in two different regimens: i) drinking of 36 g alcohol daily and ii) drinking only non-alcoholic beverages, to test whether alcohol-induced increase of HDL cholesterol can affect cholesterol efflux (CHE) from cell culture of labeled human macrophages. Alcohol consumption induced significant (p < 0.05) increases of HDL cholesterol from 1.25 +/- 0.32 to 1.34 +/- 0.38 mmol/l and Apo A1 from 1.34 +/- 0.16 to 1.44 +/- 0.19 g/l. These changes were combined with a slight increase of cholesterol efflux from 13.8 +/- 2.15 to 14.9 +/- 1.85 % (p = 0.059). There were significant correlations between individual changes of HDL-C and Apo A1 concentrations and individual changes of CHE (0.51 and 0.60, respectively). In conclusion, moderate alcohol consumption changes the capacity of plasma to induce CHE only at a border line significance. PMID- 19681658 TI - Antiatherogenic effect of simvastatin is not due to decrease of LDL cholesterol in ovariectomized golden Syrian hamster. AB - The changes of the composition of blood lipoproteins caused by menopause could also change the effect of hypolipidemic therapy. Using an experimental model we studied the changes of serum lipids and the effect of immediate or delayed treatment with simvastatin on atherosclerosis after surgical menopause. Female golden Syrian hamster aged 6 months were fed hypercholesterolemic diet during the whole study. Atherosclerotic changes in thoracic and abdominal aortas were assessed by stereomicroscopic method after 12 weeks. Four experimental groups were studied: sham-operated animals (n = 5), ovariectomized animals (n = 9), ovariectomized animals treated for 12 weeks (n = 10), and ovariectomized animals treated 4 weeks after ovariectomy for 8 weeks (n = 9). The dose of simvastatin was 10 mg/kg of body weight. After 12 weeks, ovariectomized animals had tenfold higher concentration of triglycerides in LDL fraction and significantly higher prevalence of atherosclerosis than animals without ovariectomy. Treatment with simvastatin substantially decreased the prevalence of atherosclerotic changes, but otherwise did not change individual serum lipids including LDL cholesterol. However, it improved proportions of pro- and antiatherogenic serum lipids mainly by the increase of HDL cholesterol. The timing of simvastatin treatment had no significant effect on atherosclerotic changes or lipid parameters. Simvastatin treatment partly prevented atherosclerotic changes induced by ovariectomy. This effect was not mediated by decrease of LDL cholesterol, but by increase in HDL cholesterol. PMID- 19681659 TI - The effects of short-term norepinephrine up-titration on hemodynamics in cardiogenic shock. AB - A higher mean arterial pressure (MAP) achieved by norepinephrine up-titration may improve organ blood flow in critically ill, whereas norepinephrine-induced afterload rise might worsen myocardial function. Our aim was to assess the effects of norepinephrine dose titration on global hemodynamics in cardiogenic shock. We prospectively evaluated 12 mechanically ventilated euvolemic patients (aged 67 +/- 12 years) in cardiogenic shock (10 patients acute myocardial infarction, 1 patient dilated cardiomyopathy, 1 patient decompensated aortic stenosis). Hemodynamic monitoring included arterial and Swan-Ganz catheters. The first data were obtained at MAP of 65 mm Hg, then the norepinephrine dose was increased over 40 min to achieve MAP of 85 mm Hg. Finally, the norepinephrine dose was tapered over 40 min to achieve MAP of 65 mm Hg. Norepinephrine up titration increased MAP to the predefined values in all patients with concomitant mild increase in filling pressures and heart rate. Systemic vascular resistance increased, whereas cardiac output remained unchanged. During norepinephrine down titration, all hemodynamic parameters returned to baseline values. We observed no changes in lactate levels and mixed venous oxygen saturation. Our data suggest that short-term norepinephrine dose up-titration in cardiogenic shock patients treated or pretreated with inotropes was tolerated well by the diseased heart. PMID- 19681660 TI - Absence of flow-mediated vasodilation in the rabbit femoral artery. AB - The purpose of this study was to determine if there is flow-mediated vasodilation of the femoral artery in response to progressive increases in flow within a physiological range observed in the in vivo experiments. Femoral artery blood flow was determined in conscious rabbits (n = 5) using chronically implanted flowprobes. Resting blood flow was 8.3 +/- 0.6 ml/min and increased to 39.9 +/- 5.4 ml/min during high intensity exercise. Femoral arteries (n = 12, 1705 +/- 43 microm outer diameter) harvested from a separate group of rabbits were mounted on cannulas and diameter was continuously monitored by video system. Functional integrity of the endothelium was tested with acetylcholine. The arteries were set at a transmural pressure of 100 mm Hg and preconstricted with phenylephrine to 73 +/- 3% of initial diameter. Using a roller pump with pressure held constant, the arteries were perfused intraluminally with warmed, oxygenated Krebs' solution (pH = 7.4) over a physiological range of flows up to 35 ml/min. As flow increased from 5 ml/min to 35 ml/min, diameter decreased significantly (p < 0.05) from 1285 +/- 58 microm to 1100 +/- 49 microm. Thus, in vessels with a functional endothelium, increasing intraluminal flow over a physiological range of flows produced constriction, not dilation. Based on these results, it seems unlikely that flow-mediated vasodilation in the rabbit femoral artery contributes to exercise hyperemia. PMID- 19681661 TI - Levels of circulating biomarkers at rest and after exercise in coronary artery disease patients. AB - As traditional risk factors are unable to fully explain the pathogenesis of coronary artery disease (CAD), novel mechanisms became a target of many investigations. Our aim was to study the response of selected markers to physical exercise. High-sensitive C-reactive protein (hs-CRP), matrix metalloproteinases 2 and 9 (MMP-2, MMP-9), advanced oxidation protein products (AOPP), soluble receptor for advanced glycation end-products (sRAGE), pregnancy-associated plasma protein A (PAPP-A), E-selectin, vascular endothelial growth factor (VEGF) and B type natriuretic peptide (BNP) levels were measured in serum of 21 CAD patients and in 22 healthy controls at rest and after exercise bicycle stress test performed up to the maximal tolerated effort. At rest, hs-CRP, AOPP, MMP-9 and BNP were significantly elevated in the CAD patients as compared with controls. In contrast, P-selectin was significantly lower in CAD patients and a tendency to lower levels of sRAGE was noted. After exercise MMP-9 and BNP, increased significantly in both groups. In conclusions, CAD patients have elevated hs-CRP, AOPP, MMP-9 and BNP--novel markers related to cardiovascular risk or left ventricular overload. MMP-9 and BNP increase significantly with exercise in both healthy individuals and CAD patients. PMID- 19681662 TI - Osteogenic cells on bio-inspired materials for bone tissue engineering. AB - This article reviews the development of artificial bone substitutes from their older single-phase forms to novel multi-phase composites, mimicking the composition and architecture of natural bone tissue. The new generation of bone implants should be bioactive, i.e. they should induce the desired cellular responses, leading to integration of the material into the natural tissue and stimulating self-healing processes. Therefore, the first part of the review explains the common principles of the cell-material interaction and summarizes the strategies how to improve the biocompatibility and bioactivity of the materials by modifying the physico-chemical properties of the material surface, such as surface chemistry, wettability, electrical charge, rigidity, microroughness and especially nanoroughness. The latter has been shown to stimulate preferentially the growth of osteoblasts in comparison with other competitive cell types, such as fibroblasts, which could prevent fibrous tissue formation upon implantation. The second more specialized part of the review deals with materials suitable for bone contact and substitution, particularly novel polymer-based composites reinforced with fibres or inorganic particles and containing bioactive components, such as crystals of hydroxyapatite or other calcium phosphates, synthetic ligands for cell adhesion receptors or growth factors. Moreover, if they are degradable, they can be gradually replaced with a regenerating tissue. PMID- 19681663 TI - ERKs and JNKs mediate hydrogen peroxide-induced Egr-1 expression and nuclear accumulation in H9c2 cells. AB - One of the most significant insults that jeopardize cardiomyocyte homeostasis is a surge of reactive oxygen species (ROS) in the failing myocardium. Early growth response factor-1 (Egr-1) has been found to act as a transcriptional regulator in multiple biological processes known to exert deleterious effects on cardiomyocytes. We thus investigated the signaling pathways involved in its regulation by H2O2. Egr-1 mRNA levels were found to be maximally induced after 2 h in H2O2-treated H9c2 cells. Egr-1 respective response at the protein level, was found to be maximally induced after 2 h of treatment with 200 microM H2O2, remaining elevated for 6 h, and declining thereafter. H2O2-induced upregulation of Egr-1 mRNA and protein levels was ablated in the presence of agents inhibiting ERKs pathway (PD98059) and JNKs (SP600125, AS601245). Immunofluorescent experiments revealed H2O2-induced Egr-1 nuclear sequestration to be also ERK- and JNK-dependent. Overall, our results show for the first time the fundamental role of ERKs and JNKs in regulating Egr-1 response to H2O2 treatment in cardiac cells at multiple levels: mRNA, protein and subcellular distribution. Nevertheless, further studies are required to elucidate the specific physiological role of Egr 1 regarding the modulation of gene expression and determination of cell fate. PMID- 19681664 TI - Collagen synthesis, nitric oxide and asymmetric dimethylarginine in diabetic subjects undergoing hyperbaric oxygen therapy. AB - The main pathological condition in patients with impaired wound healing is diabetes mellitus. These patients have significantly low circulating nitric oxide (NO) levels because the stimulatory action of insulin on NO synthesis is absent. Additionally, asymmetric dimethylarginine (ADMA), an inhibitor of NO synthase, is increased owing to the generation of oxidative stress. NO was thought to contribute to wound healing. Hyperbaric oxygen (HBO) treatment is generally used in order to accelerate the healing of wounds. The aim of this study was to determine the changes in plasma procollagen type I and III N-terminal peptides (PINP and PIIINP), total nitrite/nitrate (NO(x)) and ADMA levels; and to evaluate their relation to healing during the HBO treatment of foot ulcers. Data obtained from 18 diabetic patients before and after the HBO therapy were compared statistically by the Wilcoxon test. NO(x) was increased in 11 and ADMA was decreased in 12 patients following HBO treatment. Both PINP (32.6 +/- 29.4 microg/l vs 44.3 +/- 33.4 microg/l) and PIIINP (6.97 +/- 3.01 microg/l vs 7.92 +/ 2.49 microg/l) were significantly increased (p < 0.05). Progressive reductions were observed in wound areas, as assessed by the digital wound imaging. In 12 patients, wounds healed by 50% or higher; whereas only two subjects had minimal improvements (15% or less healing). The duration of diabetes correlated negatively with wound healing (r = -498, p < 0.05). This study suggests that increased collagen synthesis is associated with wound healing during hyperbaric oxygen therapy. Nitric oxide generation may also contribute to the healing process. PMID- 19681665 TI - Hemodynamic response to thoracoscopy and thoracotomy. AB - Operations in the pleural cavity are connected with circulatory changes in pulmonary circulation and general changes of hemodynamics. These changes are influenced by the position of patient's body on the operation table and by the introduction of artificial pneumothorax. Thoracoscopy is an advanced surgical approach in thoracic surgery, but its hemodynamic effect is still not known. The aim of the present study was to compare the hemodynamic response to surgeries carried out by open (thoracotomy - TT) and closed (thoracoscopy - TS) surgical approach. Thirty-eight patients have been monitored throughout the operation- from the introduction of anesthesia to completing the surgery. Monitored parameters were systolic blood pressure (BPs), diastolic blood pressure (BPd), O2 saturation (SaO2), systolic blood pressure in pulmonary artery (BPPAs), diastolic blood pressure in pulmonary artery (BPPAd), wedge pressure (P(W)), central venous pressure in right atrium (CVP), cardiac output (CO) and total peripheral resistance (TPR). No significant difference has been found in hemodynamic response between TT and TS groups. Significant changes of hemodynamic parameters occurring during the whole surgical procedure were detected in both technical approaches. The most prominent changes were found after the position of patients was changed to the hip position (significantly decreased BPs, BPd, MAP, SaO2 and BPPAs) and 5 min after the pneumothorax was established (restoration of the cardiac output to the initial value and significant decrease of the TPR). It can be concluded that the thoracoscopy causes almost identical hemodynamic changes like the thoracotomy. PMID- 19681666 TI - Fitting a single-phase model to the post-exercise changes in heart rate and oxygen uptake. AB - The kinetics of post-exercise heart rate (HR) and oxygen consumption (EPOC) was studied in 10 elite cyclists subjected to four laboratory cycle ergometer maximal exercises lasting 30, 90, 180 or 360 s. Heart rate and oxygen uptake (VO2) were recorded over a period of 6 min after the exercise. By applying the logit transformation to the recorded variables and relating them to the decimal logarithm of the recovery time, uniform single-phase courses of changes were shown for both variables in all subjects and exercises. This enabled computing half-recovery times (t(1/2)) for both variables. Half-time for VO2 negatively correlated with square root of exercise duration (within-subject r = -0.629, p < 0.001), the total post-exercise oxygen uptake till t(1/2) was thus constant irrespectively of exercise intensity. The method is simple and enables reliable comparisons of various modes of exercise with respect to the rate of recovery. PMID- 19681668 TI - 2DE-proteomics meta-data indicate the existence of distinct cellular stress responsive mechanisms. PMID- 19681667 TI - Microencapsulation of isoniazid in genipin-crosslinked gelatin-A-kappa carrageenan polyelectrolyte complex. AB - BACKGROUND: Microspheres of gelatin-A and kappa-carrageenan were prepared by using genipin, a naturally occurring crosslinker, and sunflower oil as reaction media. METHOD: The variations in the size of the microspheres formed by varying the amount of surfactant (0.33-1.0 g/g of polymer), polymer (1.5-3.0 g), and crosslinker (0.2-0.8 mmol) were studied by scanning electron microscopy. The encapsulation of isoniazid was carried out by absorption. The isoniazid content in the prepared microspheres was determined. The release characteristic of isoniazid was also studied at pH values 1.2 and 7.4 by using UV spectrophotometer. RESULTS: Characterization of the isoniazid-loaded microspheres was carried out by using Fourier transform infrared spectrophotometry, differential scanning calorimetry, and X-ray diffractometery. PMID- 19681669 TI - Identification of serum melanoma progression biomarkers through proteomic-based approaches. PMID- 19681670 TI - Nutrition proteomics and biomarker discovery. AB - Baukje de Roos is a principal investigator at the University of Aberdeen, Rowett Institute of Nutrition and Health. She investigates mechanisms through which dietary fats and fatty acids, and also polyphenols, affect parameters involved in the development of heart disease in vivo. This is achieved not only by measuring their effect on conventional risk markers for heart disease but also by assessing their effect on new markers that are being developed through proteomic and mass spectrometry methods. She obtained her PhD in Human Nutrition at Wageningen University, The Netherlands, in January 2000, after which she was appointed as a post-doctoral research fellow at the Department of Vascular Biochemistry, Glasgow Royal Infirmary, in collaboration with GlaxoSmithKline. In June 2001 she joined the Rowett Research Institute in Aberdeen. She is currently working for the University of Aberdeen, where her research is funded by the Scottish Government Rural and Environment Research and Analysis Directorate (RERAD). She is an active member of the European Nutrigenomics Organisation (NuGO), an EU-funded Network of Excellence, which merges the nutrigenomics activities of its 23 partners across Europe. PMID- 19681672 TI - Thyroid tumors: novel insights from proteomic studies. AB - Several genomics-based techniques have been applied in the last decade to the molecular characterization of cancer, which has led to a variety of applications suitable for improved diagnosis, prognosis and prediction of outcome to treatment. Proteomics-based approaches have also been seen as crucial to the discovery of biomarkers for early diagnosis and prognosis of tumors, as well as for a better understanding of the molecular bases of cancer. Accordingly, proteomic techniques have been used extensively for a better molecular characterization of thyroid tumors. In this field, three main directions have been preceded: first, proteomic studies of model systems; second, proteomics of thyroid tumor specimens; and third, serum proteomics. In this review, we describe the most relevant results that have been obtained for tumors derived from thyroid follicular cells using various proteomic approaches. PMID- 19681671 TI - Tackling the human adipose tissue proteome to gain insight into obesity and related pathologies. AB - Obesity is becoming an important public health problem given its strong association with insulin resistance and Type 2 diabetes. Previously considered an inert depot, fat is now regarded as a highly metabolically active tissue in many pathophysiological processes. In humans, the accumulation of omental rather than subcutaneous adipose tissue appears to be tightly linked to cardiovascular disease and other important comorbidities. Proteomics has emerged as a method for the large-scale study of proteins in biological samples, for instance, fluids, cells or tissues, which encompasses not only the identities of the proteins present, but also quantification and post-translational modification events. Human adipose tissue proteome analysis, still in its early stages, may help understand the molecular mechanisms of obesity and the role of omental fat in the pathogenesis of obesity-associated diseases. This review covers recent advances in human adipose tissue proteomics, focusing on the analysis of the omental and the subcutaneous fat. PMID- 19681673 TI - Identifying autoantibody signatures in cancer: a promising challenge. AB - Biomarkers that show high sensitivity and specificity are needed for the early diagnosis and prognosis of cancer. An immune response to cancer is elicited in humans, as demonstrated, in part, by the identification of autoantibodies against a number of tumor-associated antigen (TAAs) in sera from patients with different types of cancer. Identification of TAAs and their cognate autoantibodies is a promising strategy for the discovery of relevant biomarkers. During the past few years, three proteomic approaches, including serological identification of antigens by recombinant expression cloning (SEREX), serological proteome analysis (SERPA) and, more recently, protein microarrays, have been the dominant strategies used to identify TAAs and their cognate autoantibodies. In this review, we aim to describe the advantages, drawbacks and recent improvements of these approaches for the study of humoral responses. Finally, we discuss the definition of autoantibody signatures to improve sensitivity for the development of clinically relevant tests. PMID- 19681674 TI - Proteomic approaches in neuroblastoma: a complementary clinical platform for the future. AB - Neuroblastoma (NB) is one of the most common solid tumors of childhood and displays a remarkable diversity in both biologic characteristics and clinical outcomes. Availability of high-throughput 'omics technologies and their subsequent application towards oncology has provided insight into the complex pathways of tumor formation and progression. Investigation of NB 'omics profiles may better define tumor behavior and provide targeted therapy with the goal of improving outcomes in patients with high-risk disease. Utilization of these technologies in NB has already led to advances in classification and risk stratification. The gradual emergence of NB-directed proteomics adds a layer of intricacy to the analysis of biologic organization but may ultimately provide a better comprehension of this complex disease. In this review, we cite specific examples of how NB-directed proteomics has provided information regarding novel biomarkers and possible therapeutic targets. We finish by examining the impact of high-throughput 'omics in the field of NB and speculate on how these emerging technologies may further be incorporated into the discipline. PMID- 19681675 TI - Tissue proteomics in atherosclerosis: elucidating the molecular mechanisms of cardiovascular diseases. AB - Atherosclerosis is a disease with higher levels of mortality in developed countries. Comprehension of the molecular mechanisms can yield very useful information in clinics for prevention, diagnosis and recovery monitoring. Proteomics represents an ideal methodology for this purpose, as proteins constitute the effectors of the different biological processes running during pathogenesis. To date, studies in atherosclerosis have been mainly focused on the search for plasma biomarkers. However, tissue proteomics allows going deeper into tissue secretomes, arterial layers or particular cells of interest, which, in turn, constitutes a more direct approximation to in vivo operating mechanisms. The aim of this review is to report latest advances in tissue proteomics in atherosclerosis and related diseases (e.g., aortic stenosis and ischemic injury). PMID- 19681676 TI - Gaucher disease: a model disorder for biomarker discovery. AB - Gaucher disease is an inherited lysosomal storage disorder, characterized by massive accumulation of glucosylceramide-laden macrophages in the spleen, liver and bone marrow as a consequence of deficient activity of glucocerebrosidase. Gaucher disease has been the playground to develop new therapeutic interventions such as enzyme-replacement therapy and substrate-reduction therapy. The availability of these costly therapies has stimulated research regarding suitable biomarkers to monitor onset and progression of disease, as well as the efficacy of therapeutic intervention. Given the important role of storage cells in the pathology, various attempts have been made to identify proteins in plasma or serum reflecting the body burden of these pathological cells. In this review, the existing data regarding biomarkers for Gaucher disease, as well as the current application of biomarkers in clinical management of Gaucher patients are discussed. Moreover, the use of several modern proteomic technologies for the identification of Gaucher biomarkers is reviewed. PMID- 19681677 TI - Proteomics and liver fibrosis: identifying markers of fibrogenesis. AB - Chronic hepatic disease damages the liver and the resulting wound-healing process might lead to liver fibrosis and subsequent cirrhosis development. Fibrosis is the excessive deposition of extracellular matrix (ECM) in the tissue as consequence of chronic liver damage. The fibrotic response triggers almost all of the complications of end-stage liver disease, including portal hypertension, ascites, encephalopathy, synthetic dysfunction and impaired metabolic capacity. Thus, efforts to understand and attenuate fibrosis have direct clinical implications. Reliable, accurate, disease-specific, noninvasive biomarkers of fibrosis and fibrogenesis in order to prevent or minimize the impact of the chronic liver disease progression are a critical need. This review aims to provide an overview of the possibilities that proteome technology can offer to the knowledge, diagnosis and prognosis of liver fibrosis. PMID- 19681678 TI - Recent advances in mass spectrometry-based peptidome analysis. AB - The peptidome, which is the low-molecular-weight subset of the proteome, has attracted increasing attention in recent years. However, with the interference of high-abundance protein components in complex biological mixtures (e.g., serum), selective extraction of endogenous peptides is the first and most important step before analyzing the peptidome. A number of methods and technologies have now been developed for the selective enrichment, fractionation, quantitative analysis of the endogenous peptides and their application in the potential biomarker discovery. This review will cover the methods and technologies developed in recent years for the peptidome analysis on the selective extraction, multidimensional separation and quantitative analysis, as well as their application for clinical diagnosis and biomarker discovery. The future prospects of the peptidome are also discussed. PMID- 19681681 TI - Potential financial impact of restriction in "never event" and periprocedural hospital-acquired condition reimbursement at a tertiary neurosurgical center: a single-institution prospective study. AB - OBJECT: The Centers for Medicare and Medicaid Services (CMS) have moved to limit hospital augmentation of diagnosis-related group billing for "never events" (adverse events that are serious, largely preventable, and of concern to the public and health care providers for the purpose of public accountability) and certain hospital-acquired conditions (HACs). Similar restrictions may be applied to physician billing. The financial impact of these restrictions may fall on academic medical centers, which commonly have populations of complex patients with a higher risk of HACs. The authors sought to quantify the potential financial impact of restrictions in never events and periprocedural HAC billing on a tertiary neurosurgery facility. METHODS: Operative cases treated between January 2008 and June 2008 were reviewed after searching a prospectively maintained database of perioperative complications. The authors assessed cases in which there was a 6-month lag time to allow for completion of hospital and physician billing. They speculated that other payers would soon adopt the present CMS restrictions and that procedure-related HACs would be expanded to cover common neurosurgery procedures. To evaluate the impact on physician billing and to directly contrast physician and hospital billing impact, the authors focused on periprocedural HACs, as opposed to entire admission HACs. Billing records were compiled and a comparison was made between individual event data and simultaneous cumulative net revenue and net receipts. The authors assessed the impact of the present regulations, expansion of CMS restrictions to other payers, and expansion to rehospitalization and entire hospitalization case billing due to HACs and never events. RESULTS: A total of 1289 procedures were completed during the examined period. Twenty-five procedures (2%) involved patients in whom HACs developed; all were wound infections. Twenty-nine secondary procedures were required for this cohort. Length of stay was significantly higher in patients with HACs than in those without (11.6 +/- 11.5 vs 5.9 +/- 7.0 days, respectively). Fifteen patients required readmission due to HACs. Following present never event and HAC restrictions, hospital and physician billing was minimally affected (never event billing as percent total receipts was 0.007% for hospitals and 0% for physicians). Nonpayment for rehospitalization and reoperation for HACs by CMS and private payers yielded greater financial impact (CMS only, percentage of total receipts: 0.14% hospital, 0.2% physician; all payers: 1.56% hospital, 3.0% physician). Eliminating reimbursement for index procedures yielded profound reductions (CMS only as percentage of total receipts: 0.62% hospital, 0.8% physician; all payers: 5.73% hospital, 8.9% physician). CONCLUSIONS: The authors found potentially significant reductions in physician and facility billing. The expansion of never event and HACs reimbursement nonpayment may have a substantial financial impact on tertiary care facilities. The elimination of never events and reduction in HACs in current medical practices are worthy goals. However, overzealous application of HACs restrictions may remove from tertiary centers the incentive to treat high-risk patients. PMID- 19681682 TI - Language dominance and mapping based on neuromagnetic oscillatory changes: comparison with invasive procedures. AB - OBJECT: Event-related cerebral oscillatory changes reflect regional brain activation. In a previous study, the authors proposed a new method to determine language dominance: examine frontal oscillatory changes during silent reading by using synthetic aperture magnetometry (SAM). The authors' aims in the present study were to establish a normal template for this method, to confirm the results of their previous study with a larger patient population, and to evaluate their method with respect to language localization. METHODS: A statistical group analysis of 14 healthy volunteers was conducted to establish a normal control. Language dominance and localization were then evaluated in a larger population of 123 consecutive patients. Study participants were instructed to silently read 100 visually presented words. Using SAM, the spatial distribution of the oscillatory changes was obtained as the Student t statistic by comparing the current density for each voxel between 1 second before and 1 second after each word presentation. Group analyses of the healthy volunteers were performed using statistical nonparametric mapping. Language dominance in the patients was determined according to the laterality index (LI) calculated using peak t values of the left and right frontal desynchronizations. Language dominance was prospectively assessed, and the results were compared with those of the Wada test (63 patients). Language localization results were quantitatively compared with those of stimulation mapping (17 patients). RESULTS: Group analysis of the healthy volunteers indicated beta to low gamma band desynchronization in the left frontal area and alpha to beta desynchronization in the left parietotemporal areas. In patients, the frontal language areas were detected in 118 persons (95.9%). Lateralization of beta or low gamma desynchronization in the inferior or middle frontal gyri corresponded well with language dominance. The introduction of the LI resulted in a quantitative evaluation of language dominance, whose results were concordant with those of the Wada test in 51 (85.0%) of 60 cases. The distance between the estimated frontal language areas and stimulation-positive sites was 6.0 +/- 7.1 mm (mean +/- SD). CONCLUSIONS: This study is the first in which magnetoencephalography (MEG) was used to determine language dominance in a large population, and the results were compared with those of the Wada test. Moreover, language localization results obtained using MEG were compared with those obtained by invasive mapping. The authors' method, which is based on neuromagnetic oscillatory changes, is a new approach for noninvasively evaluating the frontal language areas, a procedure that has been problematic using MEG dipole methods. Synthetic aperture magnetometry is a noninvasive alternative to Wada testing for language dominance and helps to determine stimulation sites for invasive mapping. PMID- 19681683 TI - Subthalamic nucleus deep brain stimulator placement using high-field interventional magnetic resonance imaging and a skull-mounted aiming device: technique and application accuracy. AB - OBJECT: The authors discuss their method for placement of deep brain stimulation (DBS) electrodes using interventional MR (iMR) imaging and report on the accuracy of the technique, its initial clinical efficacy, and associated complications in a consecutive series of subthalamic nucleus (STN) DBS implants to treat Parkinson disease (PD). METHODS: A skull-mounted aiming device (Medtronic NexFrame) was used in conjunction with real-time MR imaging (Philips Intera 1.5T). Preoperative imaging, DBS implantation, and postimplantation MR imaging were integrated into a single procedure performed with the patient in a state of general anesthesia. Accuracy of implantation was assessed using 2 types of measurements: the "radial error," defined as the scalar distance between the location of the intended target and the actual location of the guidance sheath in the axial plane 4 mm inferior to the commissures, and the "tip error," defined as the vector distance between the expected anterior commissure-posterior commissure (AC-PC) coordinates of the permanent DBS lead tip and the actual AC-PC coordinates of the lead tip. Clinical outcome was assessed using the Unified Parkinson's Disease Rating Scale part III (UPDRS III), in the off-medication state. RESULTS: Twenty-nine patients with PD underwent iMR imaging-guided placement of 53 DBS electrodes into the STN. The mean (+/- SD) radial error was 1.2 +/- 0.65 mm, and the mean absolute tip error was 2.2 +/- 0.92 mm. The tip error was significantly smaller than for STN DBS electrodes implanted using traditional frame-based stereotaxy (3.1 +/- 1.41 mm). Eighty-seven percent of leads were placed with a single brain penetration. No hematomas were visible on MR images. Two device infections occurred early in the series. In bilaterally implanted patients, the mean improvement on the UPDRS III at 9 months postimplantation was 60%. CONCLUSIONS: The authors' technical approach to placement of DBS electrodes adapts the procedure to a standard configuration 1.5-T diagnostic MR imaging scanner in a radiology suite. This method simplifies DBS implantation by eliminating the use of the traditional stereotactic frame and the subsequent requirement for registration of the brain in stereotactic space and the need for physiological recording and patient cooperation. This method has improved accuracy compared with that of anatomical guidance using standard frame-based stereotaxy in conjunction with preoperative MR imaging. PMID- 19681684 TI - Response of malignant scalp dermatofibrosarcoma to presurgical targeted growth factor inhibition. AB - Dermatofibrosarcoma protuberans (DFSP) is an uncommon, locally aggressive, malignant cutaneous tumor that sparingly presents on the scalp. Dermatofibrosarcomas often result from the formation of a fusion oncogene on translocated or supernumerary ring chromosomes 17 and 22, causing the overexpression of PDGFRbeta driven by the COL1A1 promoter. Because of uncertainty surrounding appropriate treatment of aggressive scalp DFSP, the authors performed an extensive review of the available data from a MEDLINE (Ovid) search to describe the clinical presentation and treatment options for this rare tumor. Their search identified 39 different cases, including the illustrative case presented in this study. Adjuvant therapy for this malignant lesion is not universally established in the literature. In the present case, the authors successfully treated a locally invasive scalp DFSP with presurgical therapy that specifically inhibited the PDGFbeta receptor. Imatinib significantly shrank the DFSP tumor mass, reduced hypervascularity, reduced metabolic activity on PET scanning, and permitted a safe gross-total resection. Although wide excision and Mohs micrographic surgery remain the standard surgical treatments for DFSP, the authors illustrate that presurgical chemotherapeutic treatment by imatinib provides a critical adjunct to traditional therapy. PMID- 19681685 TI - Safety of magnetic resonance imaging of deep brain stimulator systems: a serial imaging and clinical retrospective study. AB - OBJECT: With the expanding indications and increasing number of patients undergoing deep brain stimulation (DBS), postoperative MR imaging is becoming even more important in guiding clinical care and practice-based learning; important safety concerns have recently emerged, however. Although phantom model studies have driven conservative recommendations regarding imaging parameters, highlighted by 2 recent reports describing adverse neurological events associated with MR imaging in patients with implanted DBS systems, the risks of MR imaging in such patients in clinical practice has not been well addressed. In this study, the authors capitalized on their large experience with serial MR imaging (3 times per patient) to use MR imaging itself and clinical outcomes to examine the safety of MR imaging in patients who underwent staged implantation of DBS electrodes for Parkinson disease, tremor, and dystonia. METHODS: Sixty-four patients underwent staged bilateral lead implantations between 1997 and 2006, and each patient underwent 3 separate MR imaging sessions subsequent to DBS placement. The first of these was performed after the first DBS placement, the second occurred prior to the second DBS placement, and third was after the second DBS placement. Follow up was conducted to examine adverse events related either to MR imaging or to DBS induced injury. RESULTS: One hundred and ninety-two MR images were obtained, and the mean follow-up time was 3.67 years. The average time between the first and second, and second and third MR imaging sessions was 19.4 months and 14.7 hours, respectively. Twenty-two MR imaging-detected new findings of hemorrhage were documented. However, all new findings were related to acute DBS insertion, whereas there were no new findings after imaging of the chronically implanted electrode. CONCLUSIONS: Although potential risks of MR imaging in patients undergoing DBS may be linked to excessive heating, induced electrical currents, disruption of the normal operation of the device, and/or magnetic field interactions, MR imaging can be performed safely in these patients and provides useful information on DBS lead location to inform patient-specific programming and practice-based learning. PMID- 19681686 TI - Deep brain stimulation for Parkinson disease. PMID- 19681687 TI - "Never events" come to neurosurgery. PMID- 19681688 TI - Onset of symptomatic hydrocephalus requiring emergency cerebrospinal fluid diversion following high-voltage electrical burn injury. AB - High-voltage electrical injuries have been reported to cause a plethora of neurological complications including cognitive, motor, and sensory deficits in an immediate or delayed fashion. In this setting, new-onset symptomatic hydrocephalus requiring CSF shunt placement has not been described. The authors present the case of an 18-year-old man who sustained a high-voltage electrical injury with a calvarial contact point that required emergency CSF diversion within hours of injury and subsequently required placement of a lumboperitoneal shunt. Management of the open calvarial wound, which required rotational flap reconstruction, and the need for ongoing CSF diversion required care and a team approach. PMID- 19681690 TI - Resistance genes traveling the microbial internet: down the drain, up the food chain? PMID- 19681689 TI - Chronic hepatitis B: back to the future with HBsAg. PMID- 19681692 TI - 10 years' experience with the pneumococcal quinolone moxifloxacin. AB - Moxifloxacin (MXF) is the latest broad-spectrum fluoroquinolone marketed worldwide. It has in vitro activity against a wide range of Gram-positive and Gram-negative pathogens including anaerobes and intracellular organisms, as well as strains resistant to beta-lactam or macrolide antibiotics. For relevant respiratory pathogens, MXF attains the threshold values of pharmacodynamic indices predictive of clinical efficacy and minimization of resistance development. On the other hand, due to its limited activity against Pseudomonas aeruginosa, it is less suitable for 'late-onset' nosocomial infections. In clinical trials, it has been found to be at least as effective and safe as comparators, while often showing higher bacteriological success rates. In some randomized studies MXF has shown superiority over comparator regimens in the treatment of patients with community-acquired pneumonia and acute bacterial exacerbations of chronic bronchitis. A consistent observation in many clinical trials of respiratory tract infections is the early onset of effect and a faster resolution of symptoms compared with standard therapy, possibly resulting from its fast distribution into tissue and high bactericidal activity leading to more rapid bacterial eradication. Although originally developed for respiratory tract infections, MXF over the years as been shown to be effective, and consequently received approval for additional indications. PMID- 19681694 TI - Therapeutic strategies for Shiga toxin-producing Escherichia coli infections. AB - Shiga toxin (Verocytotoxin)-producing Escherichia coli (STEC or VTEC) causes serious gastrointestinal infections in humans, including diarrhea and hemorrhagic colitis, and may lead to life-threatening sequelae such as the hemolytic uremic syndrome (HUS). The triennial 'VTEC' meetings provide a multidisciplinary forum for presentation of the latest research on all aspects of STEC, with sessions addressing epidemiology of human disease, animal reservoirs and transmission of STEC via the food chain, mechanisms of pathogenesis and host response, and control and prevention strategies. Management of patients with STEC disease can be challenging, particularly since conventional antibiotic therapy is contraindicated because it is believed to increase the risk of complications by promoting release of Shiga toxin by STEC in the gut. Accordingly, this report will focus on papers presented at the meeting that addressed development of alternative therapeutic strategies. PMID- 19681693 TI - Artemether-lumefantrine: an oral antimalarial for uncomplicated malaria in children. AB - Artemether-lumefantrine (AL; Coartem, Riamet) is the first fixed-dose artemisinin combination therapy (ACT) regimen to be manufactured under Good Manufacturing Practice conditions, and is the most widely adopted ACT regimen used in malaria control programs. AL is approved for the treatment of uncomplicated malaria in adults, children and infants, and as treatment of uncomplicated malaria in nonimmune travelers returning from malarious areas. AL is efficacious for treating uncomplicated malaria in children and the frequency of associated adverse events is not higher than other available ACT regimens. In this review, available evidence on efficacy and safety of AL in the treatment of uncomplicated malaria, with emphasis on children where appropriate, and focusing on characteristics that are potentially important for malaria control policy decisions, are presented and discussed. PMID- 19681695 TI - One step closer to universal influenza epitopes. AB - Evaluation of: Sui J, Hwang WC, Perez S et al. Structural and functional bases for broad-spectrum neutralization of avian and human influenza A viruses. Nat. Struct. Mol. Biol. 16(3), 265-273 (2009). The continuous antigenic drifts and occasional antigenic shifts enable human influenza viruses to escape the human immune system. Moreover, the frequent occurrence of human H5N1-infected cases and the recent emergency of a novel swine-like human H1N1 influenza virus further reiterate the risk of the introduction of a new pandemic strain to humans through in toto transfer of an animal influenza virus. The discovery of neutralizing antibodies that are broadly reactive with multiple influenza subtypes is therefore extremely important for the influenza pandemic preparedness, for use either for therapeutic purposes or as the basis of vaccine development. Here, we review some of the recent developments in this area. PMID- 19681698 TI - Hemolytic uremic syndrome: pathogenesis and update of interventions. AB - The typical form of hemolytic uremic syndrome (HUS) is the major complication of Shiga toxin-producing Escherichia coli infections. HUS is a critical health problem in Argentina since it is the main cause of acute renal failure in children and the second cause of chronic renal failure, accounting for 20% of renal transplants in children and adolescents in Argentina. Despite extensive research in the field, the mainstay of treatment for patients with HUS is supportive therapy, and there are no specific therapies preventing or ameliorating the disease course. In this review, we present the current knowledge about pathogenic mechanisms and discuss traditional and innovative therapeutic approaches, with special focus in Argentinean contribution. The hope that a better understanding of transmission dynamics and pathogenesis of this disease will produce better therapies to prevent the acute mortality and the long-term morbidity of HUS is the driving force for intensified research. PMID- 19681700 TI - TB diagnostic tests: how do we figure out their costs? AB - Tuberculosis continues to be a major global health problem. Lack of accurate, rapid and cost-effective diagnostic tests poses a huge obstacle to global TB control. While several new diagnostic tools are being developed and evaluated for TB, it is important that new tools are introduced for widespread use only after careful validation of accuracy, impact as well as cost-effectiveness in real world settings. While there are large numbers of studies on the accuracy of TB diagnostic tests, there are few studies that are focused on cost and cost effectiveness. There are currently no widely accepted standards on how to evaluate costs of a TB test. In this review, we describe the basic approach for computing the costs of TB diagnostic tests, and provide templates for various data elements and parameters that go into the costing analysis. We hope this will pave the way for a standardized methodology for costing of TB diagnostic tests. Such a tool would enable improved and more generalizable costing analyses that can provide a strong foundation for more sophisticated economic analyses that evaluate the full economic and epidemiological impact resulting from the implementation and routine use of performance-verified new and innovative diagnostic tools. This, in turn, will facilitate evidence-based adoption and use of new diagnostics, especially in resource-limited settings. PMID- 19681701 TI - Enterovirus 71: epidemiology, pathogenesis and management. AB - Enterovirus 71 (EV71) has emerged as a major cause of neurological threat in the world following the eradication of poliovirus. Most EV71 infections commonly result in hand-foot-mouth disease or herpangina, and some cases are associated with brainstem encephalitis and acute flaccid paralysis. Mortality was high in EV71 brainstem encephalitis complicated with pulmonary edema, particularly in children below 5 years of age. Destruction of vasomotor in the brainstem by EV71 produces autonomic nervous system dysregulation prior to the pulmonary edema. The pulmonary edema is the result of increased pulmonary vascular permeability caused by the direct brainstem lesions and/or a systemic inflammatory response syndrome produced by the release of cytokines and chemokines. There is currently no specific antiviral agent to treat or vaccine to prevent EV71 diseases. Treating severe EV71 brainstem encephalitis patients with intravenous IgG and milrinone is associated with significantly decreased mortality by attenuated sympathetic activity and cytokine production. PMID- 19681699 TI - Current management of human granulocytic anaplasmosis, human monocytic ehrlichiosis and Ehrlichia ewingii ehrlichiosis. AB - Anaplasma phagocytophilum, Ehrlichia chaffeensis and Ehrlichia ewingii are emerging tick-borne pathogens and are the causative agents of human granulocytic anaplasmosis, human monocytic ehrlichiosis and E. ewingii ehrlichiosis, respectively. Collectively, these are referred to as human ehrlichioses. These obligate intracellular bacterial pathogens of the family Anaplasmataceae are transmitted by Ixodes spp. or Amblyomma americanum ticks and infect peripherally circulating leukocytes to cause infections that range in clinical spectra from asymptomatic seroconversion to mild, severe or, in rare instances, fatal disease. This review describes: the ecology of each pathogen; the epidemiology, clinical signs and symptoms of the human diseases that each causes; the choice methods for diagnosing and treating human ehrlichioses; recommendations for patient management; and is concluded with suggestions for potential future research. PMID- 19681704 TI - Refractory disseminated coccidioidomycosis and mycobacteriosis in interferon gamma receptor 1 deficiency. AB - Severe coccidioidomycosis is rare, and specific genetic susceptibility to the disease remains unidentified. We describe a patient with disseminated recalcitrant coccidioidomycosis with autosomal dominant interferon-gamma receptor 1 deficiency caused by a heterozygous IFNGR1 818del4 mutation. Therefore, the interleukin-12/interferon-gamma axis appears to be critical for control of coccidioidomycosis. PMID- 19681702 TI - Autophagy as an antimicrobial strategy. AB - Autophagy is a process of lysosomal degradation that was originally described as a cellular response to adapt to a lack of nutrients and to enable the elimination of damaged organelles. Autophagy is increasingly recognized as a process that is also involved in innate and adaptive immune responses against pathogens. Studies on the regulation of autophagy have uncovered components of the autophagic cascade that can be manipulated pharmacologically. Approaches to modulate autophagy may result in novel strategies for the treatment and prevention of various infections. PMID- 19681705 TI - Human paragonimiasis in North America following ingestion of raw crayfish. AB - Paragonimiasis (human infections with the lung fluke Paragonimus westermani) is an important public health problem in parts of Southeast Asia and China. Paragonamiasis has rarely been reported from North America as a zoonosis caused by Paragonimus kellicotti. Paragonimus species have complex life cycles that require 2 intermediate hosts, namely, snails and crustaceans (ie, crabs or crayfish). Humans acquire P. kellicotti when they consume infected raw crayfish. Humans with paragonimiasis usually present with fever and cough, which, together with the presentation of hemoptysis, can be misdiagnosed as tuberculosis. Only 7 autochthonous cases of paragonimiasis have been previously reported from North America. Our study describes 3 patients with proven or probable paragonimiasis with unusual clinical features who were seen at a single medical center during an 18-month period. These patients acquired their infections after consuming raw crayfish from rivers in Missouri. It is likely that other patients with paragonimiasis have been misdiagnosed and improperly treated. Physicians should consider the possibility that patients who present with cough, fever, hemoptysis, and eosinophilia may have paragonimiasis. PMID- 19681706 TI - Spontaneous control of viral replication during primary HIV infection: when is "HIV controller" status established? AB - Eight patients in the ANRS PRIMO cohort experienced early spontaneous viral control. Viral control was established a median of 6.2 months after primary human immunodeficiency virus type 1 infection and lasted a median of 4.1 years. Seven of the patients initially had detectable viral replication. For 4 patients, viral control was lost during follow-up. PMID- 19681707 TI - Toxicodynamics of itraconazole: implications for therapeutic drug monitoring. AB - We explored concentration-toxicity relationships for itraconazole among 216 patients. Logistic regression revealed a progressive increase in the probability of toxicity with increasing concentrations of itraconazole. Classification and regression tree analysis suggested that 17.1 mg/L of itraconazole (measured using a bioassay) was the concentration level at which the population of patients was separated into 2 groups, each with a high and a low probability of toxicity. PMID- 19681708 TI - Cryptococcal immune reconstitution inflammatory syndrome after antiretroviral therapy in AIDS patients with cryptococcal meningitis: a prospective multicenter study. AB - A prospective multicenter study of cryptococcal immune reconstitution inflammatory syndrome (IRIS) was conducted as a substudy of the Bacteriology and Mycology Study Group 3-01. Of 101 AIDS patients with cryptococcal meningitis who received highly active antiretroviral therapy (HAART), 13 experienced cryptococcal IRIS. No association between the timing of HAART initiation and the diagnosis of IRIS was identified. Increased baseline serum cryptococcal antigen (CrAg) titer was a risk factor for cryptococcal IRIS. PMID- 19681710 TI - Ruling out novel H1N1 influenza virus infection with direct fluorescent antigen testing. AB - We evaluated the ability of direct fluorescent antigen (DFA) influenza tests to identify novel H1N1 influenza virus. DFA results were compared with polymerase chain reaction results. The negative predictive value of DFA testing was at least 96%. Therefore, when performed on specimens of adequate quality, DFA tests can effectively rule out infection due to novel H1N1 virus. PMID- 19681711 TI - Device-related infections in home health care and hospice: infection rates, 1998 2008. PMID- 19681709 TI - Herpes simplex encephalitis during treatment with tumor necrosis factor-alpha inhibitors. AB - We report 3 cases of herpes simplex virus encephalitis in patients receiving tumor necrosis factor-alpha (TNF-alpha) inhibitors for rheumatologic disorders. Although TNF-alpha inhibitors have been reported to increase the risk of other infectious diseases, to our knowledge, an association between anti-TNF-alpha drugs and herpes simplex virus encephalitis has not been previously described. PMID- 19681713 TI - Primate veterinarians promote animal welfare, biomedical research. One-health approach bridges gap between science and medicine. PMID- 19681714 TI - Thoughts on animal models for human disease and treatment. PMID- 19681715 TI - Food animal medicine in crisis. PMID- 19681716 TI - What is your diagnosis? Subsolar abscess with secondary distal interphalangeal joint synovitis. PMID- 19681717 TI - Pathology in practice. Interstitial pneumonia with fibrosis and intranuclear inclusion bodies. PMID- 19681718 TI - Prevalence of lymphoplasmacytic synovitis in dogs with naturally occurring cranial cruciate ligament rupture. AB - OBJECTIVE: To determine the prevalence of lymphoplasmacytic synovitis (LPS) in dogs with naturally occurring cranial cruciate ligament (CCL) rupture and compare clinical, radiographic, cytologic, and histologic findings in dogs with and without LPS. DESIGN: Cross-sectional study. ANIMALS: 110 dogs with naturally occurring CCL rupture. PROCEDURES: Histologic examination of synovial biopsy specimens obtained at the time of surgical treatment was used to identify dogs with LPS. Clinical, radiographic, cytologic, and histologic findings were compared between dogs with and without LPS. RESULTS: 56 (51%) dogs had histologic evidence of LPS. There were no significant differences in age, body weight, duration of lameness, severity of lameness, severity of radiographic signs of degenerative joint disease, extent of CCL rupture (partial vs complete), or gross appearance of the medial meniscus between dogs with and without LPS. Mean tibial plateau angle was significantly lower in dogs with LPS than in dogs without LPS, and dogs with LPS were significantly more likely to have neutrophils in their synovial fluid. Lymphocytes were seen in synovial fluid from a single dog with LPS. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that LPS was common in dogs with naturally occurring CCL rupture. However, only minor clinical, radiographic, cytologic, and histologic differences were identified between dogs with and without LPS. PMID- 19681719 TI - Evaluation of hormone receptor expression for use in predicting survival of female dogs with malignant mammary gland tumors. AB - OBJECTIVE: To evaluate the prognostic potential of expression of hormone receptors in malignant mammary gland tumors of dogs. Design-Cohort study. ANIMALS: 89 female dogs with malignant mammary gland tumors and 24 female dogs with benign mammary gland tumors. PROCEDURES: Female dogs with malignant (n = 89 dogs) and benign (24) mammary gland tumors were evaluated to determine the prognostic value of the expression of estrogen receptor (ER)A or the progesterone receptor (PR), as determined by use of immunohistochemical methods. RESULTS: In this study, 68 (60.2%) and 88 (77.9%) of the 113 dogs with mammary gland tumors had expression of ERA and PR, respectively. Expression of ERA and PR was detected proportionately more frequently in benign tumors (23/24 [95.8%] and 24/24 [100%], respectively) than in malignant tumors (45/89 [50.6%] and 64/89 [71.9%]). Percentage of tumors with positive results for ERA and PR was significantly higher in tumors < 5 cm in diameter; as clinical stage I, II, or III; and without metastasis to lymph nodes or distant metastasis. However, only PR expression in tumor cells was significantly associated with 1-year survival after surgical removal of the tumor. Moreover, dogs with malignant tumors expressing ERA and PR had a significantly higher survival rate, compared with the rate for dogs with malignant tumors expressing ERA but not PR. CONCLUSIONS AND CLINICAL RELEVANCE: These findings strongly suggested that expression of PR could be used as a prognostic factor for survival, especially in female dogs with malignant mammary gland tumors with ERA expression. PMID- 19681720 TI - Long-term prospective evaluation of topically applied 0.1% tacrolimus ointment for treatment of perianal sinuses in dogs. AB - OBJECTIVE: To evaluate effectiveness of a combination of topically applied tacrolimus, orally administered prednisone, and a novel-protein diet for treatment of perianal sinuses in dogs and to monitor clinical progress and owner management of the condition for 2 years. DESIGN: Noncontrolled clinical trial. Animals-19 dogs with perianal sinuses. Procedures-Perianal sinuses were diagnosed during physical examination, and dogs were placed on a 16-week treatment protocol consisting of topically applied 0.1% tacrolimus ointment, orally administered prednisone (tapering dose), and a novel-protein diet. Metronidazole was orally administered for the first 2 weeks. Anal sacculectomy was recommended whenever anal sacs were involved. Dogs were evaluated every month for the first 4 months and then every 6 to 12 weeks for 2 years. RESULTS: Perianal sinuses resolved completely in 15 of 19 dogs during the 16 weeks. In the remaining 4 dogs, the lesions markedly improved but failed to completely resolve. Three of these had anal sac involvement, and the owner of 1 dog had complied poorly with treatment instructions. During the 2 years following treatment, all dogs were maintained on intermittently applied tacrolimus ointment, 4 dogs also received prednisone every other day, and 11 dogs remained on the novel-protein diet. At the conclusion of the study, 13 of the 15 dogs that survived to that point were free of perianal disease. CONCLUSIONS AND CLINICAL RELEVANCE: The described protocol was effective and economical for resolving perianal sinuses. Dogs maintained on intermittent medications were unlikely to redevelop lesions. When the anal sacs were involved, anal sacculectomy appeared to improve the outcome. PMID- 19681722 TI - Evaluation of vehicular trauma in dogs: 239 cases (January-December 2001). AB - OBJECTIVE: To describe a population of dogs with vehicular trauma and to determine whether age, type and severity of injury, or preexisting disease were associated with outcome. DESIGN: Retrospective case series. ANIMALS: 239 dogs evaluated at a university referral hospital after vehicular trauma over a 12 month period. PROCEDURES: Patient characteristics, including age, outcome, animal trauma triage (ATT) score, treatments performed, hospital stay, cost, and preexisting disease, were recorded from medical records of dogs that had vehicular trauma. Dogs were assigned to a young, middle-aged, or geriatric age group. Categoric and continuous variables were compared between survivors and nonsurvivors to identify possible associations. RESULTS: 239 dogs (126 males and 113 females) were evaluated following vehicular trauma during 2001: young (n = 149), middle-aged (68), and geriatric (22). The median ATT score was 3 (range, 0 to 15). Sixteen dogs had preexisting disease. Hospital stay ranged from < 1 to 28 days (median, 3 days). Cost ranged from $77 to $10,636 (median, $853). Two hundred six dogs were discharged. Twenty-six dogs were euthanatized, and 7 died. Dogs that died or were euthanatized had significantly higher ATT scores. The ATT score also was associated with a significantly higher cost of care. Dogs with multiple injuries had significantly higher ATT scores, had increased cost of care, and were significantly more likely to die or be euthanatized. CONCLUSIONS AND CLINICAL RELEVANCE: Increased injury severity in dogs was associated with increased mortality rates and higher cost of treatment. PMID- 19681723 TI - Computed tomography and low-field magnetic resonance imaging of the pituitary gland in dogs with pituitary-dependent hyperadrenocorticism: 11 cases (2001 2003). AB - OBJECTIVE: To compare the results of computed tomography (CT) and magnetic resonance imaging (MRI) of the pituitary gland in dogs with pituitary-dependent hyperadrenocorticism (PDH) caused by histologically confirmed pituitary adenoma. DESIGN: Retrospective case series. ANIMALS: 11 dogs with PDH that underwent transsphenoidal hypophysectomy. PROCEDURES: Medical records of dogs examined between January 2001 and March 2003 were reviewed. Dogs were included in this study if they had clinical signs of hypercortisolism at the time of admission (for which PDH was diagnosed) and underwent transsphenoidal hypophysectomy. Pre- and postcontrast CT and low-field MRI (0.2-Tesla magnet) were performed on the same day as surgery for each dog. RESULTS: An abnormal pituitary gland was found in 7 dogs by use of MRI and in the same 7 dogs by use of CT. Significant differences were found between postcontrast CT and MR images for height, width, and length of the pituitary gland; brain area; and thickness of the sphenoid bone. However, the pituitary gland height-to-brain area ratio determined from postcontrast CT and MR images was not significantly different. The signal-to noise ratio and contrast-to-noise ratio of pre- and postcontrast MR images were significantly higher than those of the CT images. CONCLUSIONS AND CLINICAL RELEVANCE: Low-field MRI and dynamic CT imaging of the pituitary gland provided comparable information on the presence of pituitary adenomas in dogs with PDH. PMID- 19681725 TI - Evaluation of plasma fibrinogen concentration as an indicator of physeal or epiphyseal osteomyelitis in foals: 17 cases (2002-2007). AB - OBJECTIVE: To determine whether high plasma fibrinogen concentration (> or = 900 mg/dL) is a valid indicator of physeal or epiphyseal osteomyelitis in foals. DESIGN: Retrospective case series. ANIMALS: 17 foals with physeal or epiphyseal osteomyelitis with or without septic arthritis, 17 foals with septic arthritis alone, 20 foals with non-Rhodococcus equi pneumonia, and 22 healthy foals. PROCEDURES: Medical records were reviewed for information regarding signalment and total WBC count, segmented neutrophil count, and plasma fibrinogen concentration measured when foals were initially evaluated at the hospital. Whether the foals survived to discharge from the hospital was also determined. RESULTS: Foals with physeal or epiphyseal osteomyelitis had higher plasma fibrinogen concentrations than did foals in the other 3 groups. Sixteen of 17 affected foals had values > or = 900 mg/dL. The positive predictive and negative predictive values for plasma fibrinogen concentrations between 900 and 1,500 mg/dL as an indicator of osteomyelitis were 84.2% and 98.2%, respectively. Fibrinogen concentrations of 200 to 400 mg/dL and 500 to 800 mg/dL were associated with foals not having physeal osteomyelitis. Having septic arthritis alone or non-R equi pneumonia was significantly associated with a plasma fibrinogen concentration between 500 and 800 mg/dL; however, the positive predictive value of this range as an indicator of those diseases was low. Foals with osteomyelitis had greater total WBC and segmented neutrophil counts than did foals with septic arthritis alone. CONCLUSIONS AND CLINICAL RELEVANCE: A plasma fibrinogen concentration of > or = 900 mg/dL may be useful as an indicator of physeal or epiphyseal osteomyelitis in foals. PMID- 19681726 TI - Subtotal or partial unilateral arytenoidectomy for treatment of arytenoid chondritis in five calves. AB - CASE DESCRIPTION: 5 calves were evaluated for abnormal respiratory noise associated with variable degrees of respiratory distress. CLINICAL FINDINGS: Tachypnea and inspiratory dyspnea were detected at initial evaluation in all calves. Endoscopic evaluation of the upper respiratory tract revealed enlarged and immobile arytenoids. Radiographic (n = 3) and computed tomographic (1) evaluation of the laryngeal area revealed images that were indicative of a large soft tissue mass at the level of the arytenoids obstructing the rima glottis. A presumptive diagnosis of arytenoid chondritis was made. TREATMENT AND OUTCOME: A tracheostomy tube was placed in all calves. Medical treatment (with antimicrobials and anti-inflammatory drugs) was attempted in 4 calves after initial evaluation. Unilateral arytenoidectomy via a laryngotomy was performed under general anesthesia in all calves. Dysphagia and coughing were the most frequent postoperative complications. Three calves survived at least 6 months after the procedure. One calf died of a perforated abomasal ulcer 3 months after the surgery. Another calf died suddenly 1 month after the surgery of an undetermined cause. CLINICAL RELEVANCE: Unilateral arytenoidectomy was a viable surgical treatment for arytenoid chondritis in calves. Further research in a larger number of affected cattle is needed to determine the advantages or disadvantages of this procedure over other surgical techniques. PMID- 19681727 TI - Hematologic and plasma biochemical findings in cold-stunned Kemp's ridley turtles: 176 cases (2001-2005). AB - OBJECTIVE: To document hematologic and plasma biochemical values for a large number of cold-stunned Kemp's ridley turtles at the beginning of rehabilitation, to investigate differences in hematologic and plasma biochemical values of turtles that ultimately survived versus those that died, and to compare values of survivors during convalescence with initial values obtained at the time of admission. DESIGN: Retrospective case series. ANIMALS: 176 stranded, cold-stunned Kemp's ridley turtles hospitalized between 2001 and 2005. PROCEDURES: Hematologic and plasma biochemical values obtained at the time of admission were compared retrospectively for turtles that died versus turtles that survived. Initial results for survivors were compared with convalescent results obtained later in rehabilitation. RESULTS: Turtles that died had significantly greater plasma concentrations of sodium, chloride, potassium, calcium, phosphorus, and uric acid than did turtles that survived. For survivors, values obtained during convalescence for BUN concentration and plasma calcium concentration were significantly greater than initial values obtained at the time of admission, whereas values obtained during convalescence for glucose, sodium, and uric acid concentrations were significantly lower than initial values. CONCLUSIONS AND CLINICAL RELEVANCE: Cold-stunned Kemp's ridley turtles may be affected by electrolyte derangements, dehydration, and decreased renal function. Hematologic and plasma biochemical evaluation of such turtles provided useful clinical and prognostic information during the rehabilitation process. PMID- 19681728 TI - A phenotypic comparison of proteoglycan production of intervertebral disc cells isolated from rats, rabbits, and bovine tails; which animal model is most suitable to study tissue engineering and biological repair of human disc disorders? AB - The nucleus pulposus (NP) of the intervertebral disc in cattle and humans shows the most dramatic changes with aging of any cartilaginous tissue. In humans, notochordal cells disappear from the NP and are replaced with chondrocytic cells by adolescence. However, notochordal cells of the NP persist into adult life in some species, such as rats and rabbits. Therefore, comparison of the metabolic activity of notochordal and nonnotochordal cells is considered to be important for determining the type of cell to use for transplantation to regenerate intervertebral discs. In this study, we investigated the notochordal NP cells of rats and rabbits, as well as nonnotochordal (chondrocyte-like) bovine NP cells, in a three-dimensional culture system to examine whether proteoglycan metabolism varied among these three cell types. As a result, bovine NP cells produced around 0.18 mg/mL of glycosaminoglycan after culture for 5 days, while rat and rabbit NP cells produced about four and two times more glycosaminoglycan than bovine cells, respectively. In conclusion, this study demonstrated marked differences of energy metabolism and production of matrix components between notochordal and nonnotochordal NP cells. Animals with notochordal cells in the NP, such as rats and rabbits, may not provide good models for investigation of biological repair and tissue engineering for human disc disorders. PMID- 19681729 TI - Two-year inhalation study of carcinogenicity and chronic toxicity of 1,4-dioxane in male rats. AB - Carcinogenicity and chronic toxicity of 1,4-dioxane were examined by inhalation exposure of 50 male F344 rats to 1,4-dioxane vapor at 0 (clean air), 50, 250, or 1250 ppm (v/v) for 6 h/day, 5 days/wk, and 104 wk. Survival rates of 250 and 1250 ppm-exposed groups were decreased near the end of the 2-yr exposure period, due probably to the occurrence of malignant tumors. A statistically significant but marginal decrement of terminal body weight (<10%) was found in the 1250 ppm exposed group, suggesting slight systemic toxicity. Significant changes in plasma levels of AST, ALT, ALP, and gamma-GTP and relative weight of the liver occurred in the 1250 ppm-exposed group. Dose-dependent and statistically significant increases in incidences of nasal squamous cell carcinomas, hepatocellular adenomas, and peritoneal mesotheliomas were found primarily in the 1250 ppm exposed group. The incidences of renal cell carcinomas, fibroadenomas in the mammary gland, and adenomas in the Zymbal gland were also increased dose dependently. Preneoplastic lesions occurred in the nasal cavity and liver of the 1,4-dioxane-exposed groups. As nonneoplastic lesions, the significantly increased incidences of nuclear enlargement, atrophy, and respiratory metaplasia in the nasal cavity were noted at 50 ppm and above. A LOAEL (lowest observed adverse effect level) was determined at 50 ppm for the nasal endpoint of general chronic toxicity. This study provides clear evidence of carcinogenicity for 1,4-dioxane in male rats. A cytotoxic-proliferative and in vivo genotoxic mode of action is suggested to operate in 1,4-dioxane-induced carcinogenesis. PMID- 19681730 TI - Effect of polymerized toner on rat lung in chronic inhalation study. AB - In order to evaluate the chronic effect of polymerized toner particles on the lung, inflammation- and fibrosis-related genes were analyzed and 8 hydroxydeoxyguanosine (8-OHdG) was examined by using the lung tissue of rats subjected to 24 months of toner inhalation exposure. Wistar female rats were divided into four groups (5 weeks old, 30 rats in each): the high concentration exposure group (16.3 +/- 0.6 mg/m(3)), the medium concentration exposure group (4.4 +/- 0.3 mg/m(3)), the low concentration exposure group (1.6 +/- 0.2 mg/m(3)), and the control group (clean air). The material used was black toner, and its aerodynamic diameter in the exposure chamber was 3.0 microm. The rats were exposed to the material for 24 months (6 hours/day, 5 days/week) and dissected after the exposure period. RNA was extracted from one lung and the gene expression related to inflammation and fibrosis. Matrix metalloproteinase-2 (MMP 2), tissue inhibitor of metalloproteinase-2 (TIMP-2), and type I collagen were analyzed according to the ratio of each gene/beta-actin. Also, 8-OHdG level in the lung tissue was measured by HPLC with an electrochemical detector. Small fibrotic foci were found in the toner exposed groups; however, progressive or irreversible fibrosis was not found. The incidence of small fibrotic foci and cell aggregation increased in a dose-dependent manner. There were no significant differences of expression of MMP-2, TIMP-2, and type I collagen between the control group and each exposed group. Lung tumors did not develop in each group. A significant production of 8-OHdG was not observed in the toner exposed groups. In conclusion, toner produced by polymerization was not associated with evidence of carcinogenesis in this experiment. PMID- 19681731 TI - Early suppression of NFkappaB and IL-8 in bronchial epithelium after ozone exposure in healthy human subjects. AB - Exposure to elevated concentrations of ozone, a common air pollutant, has been associated with numerous adverse health effects. We have previously reported the time-course of ozone-induced airway inflammation, demonstrating an early up regulation of vascular endothelial adhesion molecules in bronchial mucosa at 1.5 hours, followed by a neutrophilic infiltration 6 hours after exposure to 0.2 ppm ozone. We hypothesized that the neutrophilic infiltration in the bronchial mucosa would reflect an early increase in bronchial epithelial expression of redox sensitive transcription factors and kinases regulating neutrophil chemoattractant expression. To test this hypothesis, endobronchial biopsies were obtained from healthy human subjects (n = 11) 1.5 hours after 0.2 ppm of ozone and filtered air exposures (lasting for 2 hours) and stained for mitogen-activated protein kinases (MAPKs), transcription factors, and neutrophil chemoattractants. Total epithelial staining was quantified, as well as the extent of nuclear translocation. Contrary to expectation, ozone significantly suppressed total and nuclear expression of nuclear factor kappaB (NFkappaB) in bronchial epithelial cells (p = 0.02 and p = 0.003 respectively). Similarly, the total staining for phosphorylated C-jun was suppressed (p = 0.021). Expression of interleukin 8 (IL-8) in the bronchial epithelium was likewise decreased after ozone (p = 0.018), while GRO-alpha, ENA 78, C-fos, p-p38, p-JNK, and p-ERK stainings were unchanged. These data suggest that the redox-sensitive NFkappaB and activator protein 1 (AP-1) pathways within the human bronchial epithelium do not seem to be involved in the early inflammatory cell recruitment pathways in healthy subjects exposed to ozone. PMID- 19681732 TI - Long-term responses of canine lungs to acidic particles. AB - Sixteen beagle dogs were housed in four large chambers under minimum restraint. They were exposed for 16 months to clean air and individual baseline data of markers were obtained. For 13 months, eight dogs were further exposed to clean air and eight dogs for 6 h/d to 1-microm MMAD (mass median aerodynamic diameter) acidic sulfate particles carrying 25 micromol H(+) m(-3) into their lungs. To establish functional responses (lung function, cell and tissue integrity, redox balance, and non-specific respiratory defense capacity), each exposed animal served as its own control. To establish structural responses, the eight non exposed animals served as controls. Acidic particles were produced by nebulization of aqueous sodium hydrogen sulfate at pH 1.5. Only subtle exposure related changes of lung function and structure were detected. A significant increase in respiratory burst function of alveolar macrophages points to a marginal inflammatory response. This can be explained by the significant production of prostaglandin E(2), activating cyclooxygenase-dependent mechanisms in epithelia and thus inhibiting lung inflammation. The non-specific defense capacity was slightly affected, giving increased tracheal mucus velocity and reduced in vivo dissolution of moderately soluble test particles. Hypertrophy and hyperplasia of bronchial epithelia were not observed, but there was an increase in volume density of bronchial glands and a shift from neutral to acidic staining of epithelial secretory cells in distal airways. The acidic exposure had thus no pathophysiological consequences. It is therefore unlikely that long-term inhalation of acidic particles is associated with a health risk. PMID- 19681733 TI - Effects of sulfur dioxide on expressions of p53, bax and bcl-2 in lungs of asthmatic rats. AB - Inhibition of cell apoptosis is an increasingly important factor in modulating airway inflammation in asthma, which is related to environmental pollutants. To investigate the effects of sulfur dioxide (SO(2)) on the mRNA and protein expressions of apoptosis-related genes in lungs from asthmatic rats, male Wistar rats were challenged by ovalbumin (OVA) or SO(2) (2 ppm) inhalation alone or together. Examinations were performed 24 h after the last treatment. The mRNA and protein levels of p53, bax, and bcl-2 were analyzed in lungs using real-time reverse transcription-polymerase chain reaction (RT-PCR) assay and Western blot analysis, respectively. The results indicated that increases of bcl-2 or decreases of p53 and bax mRNA and protein levels were not significant in lungs of rats exposed to SO(2) alone, compared with controls, but elevated or reduced levels of these genes appeared in lungs of asthmatic rats exposed to SO(2) plus OVA, compared with controls, suggesting that SO(2) exposure could result in OVA induced increases or decreases of transcription and translation levels of these apoptosis-related genes in rat lungs, and may have relations to airway inflammation in asthma. The regulation mechanism of apoptosis in asthma disease exposure to SO(2) needs further study. PMID- 19681734 TI - Gene expression profiling of maternal blood in early onset severe preeclampsia: identification of novel biomarkers. AB - AIMS: To investigate candidate genes in peripheral blood mononuclear cell (PBMC) that are associated with early onset severe preeclampsia (ES-PE) and to describe candidate genes function using microarrays and real-time polymerase chain reaction (PCR). METHODS: PBMC RNA was extracted from six patients with ES-PE and five uncomplicated pregnancies. The HG_U133 plus 2.0 Affymetrix GeneChips that represented 47,000 genes were used to measure gene expression in each sample. Significance analysis of microarray identified potential signature genes characterizing ES-PE vs. uncomplicated pregnancies. Eight genes were selected for confirmation by real-time PCR of 32 patients with ES-PE and 24 uncomplicated pregnancies, matched for maternal age, parity, race and gestational weeks. RESULTS: Using a whole-genome approach to study the molecular determinants of ES PE, 72 genes were found to be differentially expressed between cases and controls, including 38 up-regulated genes and 34 down-regulated genes in the group of ES-PE. Killer cell immunoglobulin-like receptor, three domains, long cytoplasmic tail, 2 (KIR3DL2), aldo-keto reductase family 1, member C3 (AKR1C3), churchill domain containing 1 (CHURC1), and solute carrier family 25, member 13 (SLC25A13) were validated to be down-regulated in the patients with ES-PE by real time PCR. CONCLUSIONS: Expression of genes with diverse function is associated with ES-PE risk, providing opportunities for the development of non-invasive diagnosis. PMID- 19681735 TI - Delivery mode for the extremely premature fetus: a statement of the prematurity working group of the World Association of Perinatal Medicine. AB - Recent retrospective publications have suggested that cesarean delivery may be beneficial for the extremely premature fetus. This article displays the available evidence and discusses this issue, including many aspects such as the difficulty in deciding when delivery is imminent, the negative impact on maternal morbidity and mortality and the cost to society of such a policy. The available scientific evidence does not support a recommendation for cesarean delivery for improving survival or decreasing morbidity for the extremely premature fetus. PMID- 19681736 TI - Growth of physiotherapy research funding in Australia. PMID- 19681737 TI - Rehabilitative ultrasound imaging is a valid measure of trunk muscle size and activation during most isometric sub-maximal contractions: a systematic review. AB - QUESTIONS: Is rehabilitative ultrasound imaging a valid measure of trunk muscle size and activation? Are rehabilitative ultrasound imaging measures sensitive to change? DESIGN: Systematic review of studies of criterion-related validity, construct validity, and sensitivity to change. PARTICIPANTS: People with low back pain and asymptomatic controls. OUTCOME MEASURE: Trunk muscle size and activation measured by rehabilitative ultrasound imaging, MRI and/or EMG. RESULTS: 37 studies were included. 10 studies investigated criterion-related validity and provided evidence that while ultrasound may be a valid measure of trunk muscle size, the validity of ultrasound to quantify muscle activation is context dependent, depending on the muscle involved, the contraction strategy utilised, and the intensity of muscle contraction. 23 studies provided evidence of construct validity by demonstrating the ability of ultrasound measurement to differentiate individuals in terms of back pain, anthropometry, and postures. Six studies contained a limited amount of information about sensitivity to change. CONCLUSIONS. It is valid to use rehabilitative ultrasound imaging to measure trunk muscle size and activation during most isometric sub-maximal contractions. Ultrasound measures appear sensitive to both positive and negative change. PMID- 19681738 TI - Aerobic exercise improves lung function in children with intellectual disability: a randomised trial. AB - QUESTION: In children with intellectual disability, is lung function lower than in healthy peers and does it improve with exercise? DESIGN: Randomised trial with intention-to-treat analysis and assessor blinding. PARTICIPANTS: Forty-four 12 year old children with Down syndrome or other intellectual disability with an average IQ of 42 (SD 8). INTERVENTION: The experimental group performed aerobic exercise for 30 minutes, five days per week, for eight weeks. The exercise was supervised walking, running, and cycling, with a target of moderate intensity. The control group continued usual activities and performed no specific exercise. OUTCOME MEASURES: Lung function as FEV1 and FVC in litres was measured with spirometry at baseline and after the intervention at eight weeks. Prior to the baseline measures, all participants underwent familiarisation of spirometry for one week. RESULTS: At baseline, FEV1 of the children with intellectual disability was a mean of 87% (95% CI 83 to 91) and FVC was 94% (95% CI 91 to 97) of predicted normal values. After intervention, FEV(1) had increased by 160 ml (95% CI 30 to 290) and FVC by 330 ml (95% CI 200 to 460) more in the experimental group than the control group. CONCLUSION: An 8-week program of aerobic exercise improves lung function in children with intellectual disability significantly. TRIAL REGISTRATION: ACTRN12609000365268. PMID- 19681739 TI - Quality of recalled dyspnoea is different from exercise-induced dyspnoea: an experimental study. AB - QUESTIONS: Are volunteered and endorsed descriptors of recalled breathlessness consistent with descriptors of exercise-induced breathlessness? Are volunteered and endorsed descriptors of exercise-induced breathlessness consistent? DESIGN: Within-participant, repeated measures, experimental study. PARTICIPANTS: 57 people with symptomatic chronic respiratory disease aged 71 years. INTERVENTION: There were three conditions. The first was recalled breathlessness. Two conditions of exercise-induced breathlessness were created by getting the participants to undertake the 6-min Walk Test twice (breathlessness 1 and 2). OUTCOME MEASURES: Descriptors of breathlessness were volunteered (where participants' used their own words) or endorsed (from a pre-existing list of 15 breathlessness statements). RESULTS: Emotive descriptors made up 65% of recalled descriptors compared with 11% of exercise-induced descriptors, whereas physical descriptors made up 35% of recalled descriptors compared with 89% of exercise induced descriptors. Of the 237 potential language pairs volunteered to describe recalled and exercise-induced breathlessness 1, only 27 (11%) were identical whereas of the 171 potential language pairs endorsed as describing recalled and exercise-induced breathlessness 1, 66 (39%) were identical. Of the 175 potential language pairs of descriptors volunteered to describe exercise-induced breathlessness 1 and 2, 72 (41%) were identical whereas of the 153 potential language pairs endorsed as describing exercise-induced breathlessness 1 and 2, 71 (46%) were identical. CONCLUSION: The language used to describe exercise-induced breathlessness immediately after two walking challenges was similar. However, descriptions of recalled breathlessness did not consistently match descriptions of exercise-induced breathlessness, which may reflect the different contexts under which breathlessness was recalled and induced. PMID- 19681740 TI - People who undergo revision arthroplasty report more limitations but no decrease in physical activity compared with primary total hip arthroplasty: an observational study. AB - QUESTION: Do people who have had revision arthroplasty report more limitations and less physical activity than those after primary total hip arthroplasty? Can degree of limitation and physical activity be predicted by revision arthroplasty, after adjustment for age, gender, and Charnley classification? DESIGN: Cross sectional observational study. PARTICIPANTS: 371 people after primary and 134 after revision total hip arthroplasty. OUTCOME MEASURES: Limitations were measured using the Dutch-language version of the WOMAC questionnaire and amount and intensity of physical activity was measured using the SQUASH questionnaire. RESULTS: The revision arthroplasty group reported 12% (95% CI 7 to 17) more limitations than the primary total hip arthroplasty group. They also reported 394 min/wk (95% CI 88 to 701) less physical activity and 1153 min/wk (95% CI 66 to 2241) less intensity of physical activity than the primary total hip arthroplasty group. Having had a revision arthroplasty predicted limitations regardless of whether the prediction was adjusted for age, gender, or Charnley group (B -12.1, 95% CI -17.2 to -7.0). However, having had a revision arthroplasty did not predict either amount (B -121.2, 95% CI -408.0 to 165.7) or intensity (B -912.8, 95% CI -1989.1 to 163.6) of physical activity when the prediction was adjusted for age, gender, and Charnley group. CONCLUSION: People reported more limitations after revision arthroplasty than after primary total hip arthroplasty. However, people after revision arthroplasty appeared to be equally physically active as those after primary total hip arthroplasty after adjusting for age, gender, and Charnley group. PMID- 19681741 TI - Duration of anaesthesia, type of surgery, respiratory co-morbidity, predicted VO2max and smoking predict postoperative pulmonary complications after upper abdominal surgery: an observational study. AB - QUESTION: Can the risk of developing postoperative pulmonary complications be predicted after upper abdominal surgery? DESIGN: Prospective observational study. PARTICIPANTS: 268 consecutive patients undergoing elective upper abdominal surgery who received standardised pre- and postoperative prophylactic respiratory physiotherapy. OUTCOME MEASURES: Predictors were 17 preoperative and intraoperative risk factors. A postoperative pulmonary complication was diagnosed when four or more of the following criteria were present: radiological evidence of collapse/consolidation, temperature > 38 degrees C, oxyhaemoglobin saturation < 90%, abnormal sputum production, sputum culture indicating infection, raised white cell count, abnormal auscultation findings, or physician's diagnosis of pulmonary complication. RESULTS: 35 participants (13%) developed postoperative pulmonary complications. Five risk factors predicted postoperative pulmonary complications: duration of anaesthesia (OR 4.3, 95% CI 1.7 to 10.8); surgical category (OR 2.3, 95% CI 1.1 to 4.7); current smoking (OR 2.1, 95% CI 1.0 to 4.5); respiratory co-morbidity (OR 2.1, 95% CI 1.0 to 4.4); and predicted maximal oxygen uptake (OR 2.0, 95% CI 1.0 to 4.3). A clinical rule for predicting the development of postoperative pulmonary complications predicted 82% of participants who developed complications. The odds of high risk participants developing pulmonary complications were 8.4 (95% CI 3.3 to 21.3) times that of low risk participants. CONCLUSION: This clinical rule for predicting the risk of developing postoperative pulmonary complications from five risk factors may prove useful in prioritising postoperative respiratory physiotherapy. Further research is needed to validate the rule. PMID- 19681742 TI - Diabetes predicts decreased quality of life among community-dwelling seniors undertaking progressive resistance exercise: an observational study. AB - QUESTION: What baseline characteristics predict good or poor quality of life among community-dwelling seniors undertaking a three-month progressive resistance exercise program? DESIGN: A prospective cohort observational study. PARTICIPANTS: 63 Japanese men and women over 65 years with mild disability. OUTCOME MEASURES: Health-related quality of life was measured using the Short Form 36. Predictors included age, sex, body mass index, presence of chronic diseases, frequency of exercise, number of falls, and activity limitations measured before undertaking the exercise program. RESULTS: The presence of diabetes negatively predicted a good outcome (OR 0.20, 95% CI 0.05 to 0.88) and positively predicted a poor outcome (OR 4.40, 95% CI 1.21 to 18.86) when adjusted for age, sex and body mass index. The accuracy of the prediction of a good outcome was 1.25 (LR+) and 0.44 (LR-). The accuracy of the prediction of a poor outcome was 7.15 (LR+) and 0.87 (LR-). CONCLUSION: The quality of life improved in a community-dwelling elderly population with mild disability who undertook a three-month group-based progressive resistance exercise program. However, those with diabetes mellitus were more likely to experience decreased quality of life. Therefore, health care providers need to monitor carefully participants with this disease who are undertaking progressive resistance exercise. PMID- 19681743 TI - When should an intervention be provided? PMID- 19681747 TI - Is progressive resistance exercise ineffective in increasing muscle strength in young people with cerebral palsy? PMID- 19681745 TI - Did authors draw the right conclusion? PMID- 19681749 TI - Is there sufficient evidence? PMID- 19681750 TI - Balance exercises in arthritis need to be targeted to the individual. PMID- 19681752 TI - Practical research governance 101. PMID- 19681753 TI - Complex decongestive physiotherapy for pretibial myxoedema. PMID- 19681755 TI - Impact of selenite and selenate on differentially expressed genes in rat liver examined by microarray analysis. AB - Sodium selenite and sodium selenate are approved inorganic Se (selenium) compounds in human and animal nutrition serving as precursors for selenoprotein synthesis. In recent years, numerous additional biological effects over and above their functions in selenoproteins have been reported. For greater insight into these effects, our present study examined the influence of selenite and selenate on the differential expression of genes encoding non-selenoproteins in the rat liver using microarray technology. Five groups of nine growing male rats were fed with an Se-deficient diet or diets supplemented with 0.20 or 1.0 mg of Se/kg as sodium selenite or sodium selenate for 8 weeks. Genes that were more than 2.5 fold up- or down-regulated by selenite or selenate compared with Se deficiency were selected. GPx1 (glutathione peroxidase 1) was up-regulated 5.5-fold by both Se compounds, whereas GPx4 was up-regulated by only 1.4-fold. Selenite and selenate down-regulated three phase II enzymes. Despite the regulation of many other genes in an analogous manner, frequently only selenate changed the expression of these genes significantly. In particular, genes involved in the regulation of the cell cycle, apoptosis, intermediary metabolism and those involved in Se-deficiency disorders were more strongly influenced by selenate. The comparison of selenite- and selenate-regulated genes revealed that selenate may have additional functions in the protection of the liver, and that it may be more active in metabolic regulation. In our opinion the more pronounced influence of selenate compared with selenite on differential gene expression results from fundamental differences in the metabolism of these two Se compounds. PMID- 19681754 TI - The NADPH oxidase Nox4 restricts the replicative lifespan of human endothelial cells. AB - The free radical theory of aging proposes that ROS (reactive oxygen species) are major driving forces of aging, and are also critically involved in cellular senescence. Besides the mitochondrial respiratory chain, alternative sources of ROS have been described that might contribute to cellular senescence. Noxs (NADPH oxidases) are well-known sources of superoxide, which contribute to the antimicrobial capabilities of macrophages, a process involving the prototypical member of the family referred to as Nox2. However, in recent years non-phagocytic homologues of Nox2 have been identified that are involved in processes other than the host defence. Superoxide anions produced by these enzymes are believed to play a major role in signalling by MAPKs (mitogen-activated protein kinases) and stress-activated kinases, but could also contribute to cellular senescence, which is known to involve oxygen radicals. In HUVECs (human umbilical vein endothelial cells), Nox4 is predominantly expressed, but its role in replicative senescence of HUVECs remains to be elucidated. Using shRNA (small-hairpin RNA)-mediated knockdown of Nox4, implicating lentiviral vectors, we addressed the question of whether lifelong depletion of Nox4 in HUVECs would influence the senescent phenotype. We found a significant extension of the replicative lifespan of HUVECs upon knockdown of Nox4. Surprisingly, mean telomere length was significantly reduced in Nox4-depleted cells. Nox4 depletion had no discernable influence on the activity of MAPKs and stress-activated kinases, but reduced the degree of oxidative DNA damage. These results suggest that Nox4 activity increases oxidative damage in HUVECs, leading to loss of replicative potential, which is at least partly independent of telomere attrition. PMID- 19681756 TI - Development of an unsteady-state model for a biological system in miniaturized bioreactors. AB - In the present study, the special shake flasks, so-called ventilation flasks, are equipped with oxygen sensors and then an unsteady-state gas transfer model for shake flasks was developed and experimentally investigated for a wide range of gas transfer resistances (kplug). For the validation of our unsteady-state model to simulate the gas transfer in a biological system in the ventilation flasks, a strain of Corenobacterium glutamicum DM1730 was used as a model organism. For further easy processing, the resulting total mass-transfer resistance (kplug) is described as a function of the mass flow through the sterile plug (OTRplug) by an empirical equation. This equation is introduced into a simulation model that calculates the gas partial pressures in the headspace of the flask. Additionally, the gas transfer rates through the sterile closure and gas/liquid interface inside the flask are provided. This unsteady-state model would be a very useful method for scaling up from a shake flask to a fermentor; comparing the results of the gas concentration in the gas phase, there is good agreement between the introduced unsteady-state model and experimental results for the biological system. PMID- 19681757 TI - In silico analysis of neuregulin 1 evolution in vertebrates. AB - NRG1 (neuregulin 1) belongs to the NRG family of EGF (epidermal growth factor) like signalling molecules involved in cell-cell communication during development and disease. It plays important roles in the developing tissues of the nerves, heart and mammary glands. Particularly in neurobiology, NRG1 signalling is associated with synaptic transmission, myelination of Schwann cells and the human disease of schizophrenia. Many different isoforms of NRG1 make the molecule highly sophisticated in biological activities and a great diversity of in vivo functions. The nervous system is a common trait in all bilateria (higher animals), but based on the BLAST information from the currently available databases it appears that NRG1 orthologues can only be identified in vertebrates. The gene was analysed in silico for type I-IV CDSs (coding sequences) from ten vertebrate genomes. The gene loci, structures of coding-intronic sequences, ClustalW program analyses, phylogenetic trees and conserved motifs in ecto- and cyto-plasmic domains were analysed and compared. Here, we conclude that non mammalian vertebrates mainly carry type I (may have evolved a spacer different from mammalian isoforms), II and III NRG1s. The type IV NRG1 N-terminal CDSs can be identified from most of the mammalian genomes studied; however, the corresponding rodent sequences lack the start codon. The evolutionary conservation of a CDS59-CDS24-CDS103 domain, intracellular phosphorylation sites and bipartite nuclear localization signals is of physiological significance. PMID- 19681758 TI - Skeletal muscle dysfunction in COPD: clinical and laboratory observations. AB - COPD (chronic obstructive pulmonary disease), although primarily a disease of the lungs, exhibits secondary systemic manifestations. The skeletal muscles are of particular interest because their function (or dysfunction) not only influences the symptoms that limit exercise, but may contribute directly to poor exercise performance. Furthermore, skeletal muscle weakness is of great clinical importance in COPD as it is recognized to contribute independently to poor health status, increased healthcare utilization and even mortality. The present review describes the current knowledge of the structural and functional abnormalities of skeletal muscles in COPD and the possible aetiological factors. Increasing knowledge of the molecular pathways of muscle wasting will lead to the development of new therapeutic agents and strategies to combat COPD muscle dysfunction. PMID- 19681759 TI - Polymerase chain reaction confirmed by immunohistochemistry: a two-pronged diagnostic approach in endophthalmitis. PMID- 19681760 TI - Complications of intrastromal corneal ring segment implantation using a femtosecond laser for channel creation: a survey of 850 eyes with keratoconus. AB - PURPOSE: To report complications after the implantation of intrastromal corneal ring segments (Keraring; Mediphacos, Belo Horizonte, Brazil) in keratoconic eyes using a femtosecond laser (IntraLase; Advanced Medical Optics, Santa Ana, California, USA) for channel creation. METHODS: Retrospective chart review of 531 patients (850 eyes) who underwent Keraring insertion using a femtosecond laser for channel creation. Intraoperative and postoperative complications were recorded. RESULTS: Intraoperatively, there were 22 (2.7%) cases of incomplete channel formation. The rest of the intraoperative complications were galvo lag error (system malfunction) [five eyes (0.6%)], endothelial perforation [five eyes (0.6%)] and incorrect entry of the channel [two eyes (0.2%)]. Postoperatively, there were 11 (1.3%) cases of segment migration, two (0.2%) cases of corneal melting and one (0.1%) case of mild infection. The overall complication rate was 5.7% (49 cases out of 850 eyes). CONCLUSION: In this study, intracorneal ring segment implantation using a femtosecond laser for channel creation was related to a number of complications. The most common complications were incomplete channel creation (intraoperatively) and segment migration (postoperatively). PMID- 19681761 TI - Liposomes as an ocular delivery system of fluconazole: in-vitro studies. AB - PURPOSE: To study the clinical effect of topical controlled-release ophthalmic fluconazole liposomal formulation and to compare its effect with fluconazole solution in a reproducible model of Candida keratitis in rabbits. METHODS: Forty adult rabbits were included in this study. Right eyes were inoculated with freshly prepared Caindida albicans strain no. 4925 and showed signs of infected keratitis. The rabbits were divided randomly into two groups: in the first group (18 rabbits) the right eyes received fluconazole solution, while in the second group (22 rabbits) the right eyes received fluconazole-loaded liposomes. The rabbits' eyes were examined daily over a 21-day period and results were recorded. RESULTS: Rabbits infected with C. albicans responded better and showed more improvement in terms of size of ulcer and hypopyon using fluconazole-loaded liposomal formulae than using fluconazole solution. In the first group (solution), nine rabbits' cornea showed complete healing (50%) at the end of third week while in group 2 (liposome), 19 rabbits' cornea showed complete healing (86.4%) at equal duration. These results were statistically significant. CONCLUSION: Therapy with topical liposomal fluconazole (2 mg/ml) was successful in eliminating experimental C. albicans infection of the rabbit cornea and was superior to fluconazole solution. PMID- 19681762 TI - The North Jutland County Diabetic Retinopathy Study (NCDRS). AB - PURPOSE: This study set out to map the associations between retinal lesions, visual acuity (VA) and the presence of clinically significant macular oedema (CSMO) in diabetes subjects. METHODS: This cross-sectional study comprised 656 type 1 and 328 type 2 diabetes subjects undergoing retinopathy screening in the County of North Jutland, Denmark. Numbers of specific retinal lesions were quantified from retinal photographic recordings. Associations between CSMO, number of specific retinal lesions and VA were established. The percentages of eyes with CSMO ascribed to retinal lesions were calculated. RESULTS: The presence of CSMO, number of specific retinal lesions and VA were all significantly associated. The parameter with the highest statistical association with CSMO measured by Spearman's correlation coefficient was hard exudates (type 1: 0.524; type 2: 0.715), followed by microaneurysms (type 1: 0.298; type 2: 0.508), retinal haemorrhages (type 1: 0.227; type 2: 0.595), cottonwool spots (type 1: 0.207; type 2: 0.259) and VA (type 1: - 0.137; type 2: - 0,175). CONCLUSIONS: All retinal lesions are significantly associated with CSMO and together can predict for up to 42.3% (in type 1 diabetes) and 64.3% (in type 2 diabetes) of CSMO cases. PMID- 19681763 TI - Dystrophia Smolandiensis: a novel morphological picture of recurrent corneal erosions. AB - PURPOSE: The aim of this study was to describe morphological changes in Dystrophia Smolandiensis, a corneal disease that is characterized by recurrent corneal erosive episodes and the formation of central corneal keloid-like opacities in approximately half of those affected. METHODS: The corneas of seven affected individuals were examined using in-vivo confocal microscopy. Specimens of one primary corneal graft, one regraft and one biopsied keloid-like region- all obtained from members of a large family with the disease--were re-examined with a light microscope. Sections were stained with Congo red and analysed immunohistochemically for fibronectin and S100A4. RESULTS: Light microscopic examination revealed epithelial hyperplasia, absence of Bowman's layer and subepithelial fibrosis. Fibronectin was expressed in the area of subepithelial fibrosis, and the keratocytes in this area generally expressed S100A4. The biopsy specimen stained positive for Congo red, suggesting an amyloid deposit. In-vivo confocal microscopy confirmed epithelial abnormalities, loss of Bowman's layer and significant alterations of the subbasal nerve plexus in affected individuals. CONCLUSION: The morphological picture in Dystrophia Smolandiensis is novel for a condition dominated by recurrent corneal erosions at the clinical level. Although no single morphological feature unique to the disease could be found, the general morphological pattern of pathology (true keloid formation, absence of Bowman's layer, subepithelial fibrosis and abnormal subbasal nerves) probably reflects a novel phenotypic expression of the healing response to recurrent erosion of the corneal epithelium. However, the pathogenesis of Dystrophia Smolandiensis remains to be elucidated fully. PMID- 19681764 TI - Use of the retinal vessel analyzer in ocular blood flow research. AB - The present article describes a standard instrument for the continuous online determination of retinal vessel diameters, the commercially available retinal vessel analyzer. This report is intended to provide informed guidelines for measuring ocular blood flow with this system. The report describes the principles underlying the method and the instruments currently available, and discusses clinical protocol and the specific parameters measured by the system. Unresolved questions and the possible limitations of the technique are also discussed. PMID- 19681765 TI - The effect of ginkgo biloba on the rat retinal ganglion cell survival in the optic nerve crush model. AB - PURPOSE: To investigate the effect of ginkgo biloba on the retinal ganglion cell survival in a rat optic nerve crush model. METHODS: Twenty-four Sprague-Dawley rats were divided randomly into a study group of 12 animals receiving intraperitoneal injections of ginkgo biloba and a control group of 12 animals receiving intraperitoneal saline injections. All injections were performed 1 hr before the optic nerve crush and daily afterwards. For each animal, the right optic nerve was crushed closely behind the globe for 60 seconds using a microclip with 40 g power. The left optic nerve was kept intact. At 23 days after the optic nerve crush, the retinal ganglion cells were labelled retrogradely by injecting 3% fluorogold into both sides of the superior colliculus of the brain. At 4 weeks after the optic nerve crush, the animals were killed. Photographs taken from retinal flat mounts were assessed for the number and density of the retinal ganglion cells. RESULTS: The survival rate, defined as the ratio of the retinal ganglion cell density in the right eye with the optic nerve crush divided by the retinal ganglion cell density in left eye without an optic nerve trauma, was significantly (p=0.035) higher in the study group with ginkgo biloba than in the control group (60.0+/-6.0% versus 53.5+/-8.0%). CONCLUSION: The results suggest that intraperitoneal injections of a ginkgo biloba extract given prior to and daily after an experimental and standardized optic nerve crush in rats were associated with a higher survival rate of retinal ganglion cells. PMID- 19681766 TI - Association between peer relationship problems and childhood overweight/obesity. AB - AIMS: To assess the association between peer relationship problems and childhood overweight and obesity. METHODS: Data on 4718 preschool children were obtained at the obligatory school entry health examination in Bavaria. Parentally reported peer relationship problems ('normal', 'borderline' or 'abnormal') were assessed from the Strengths and Difficulties Questionnaire. Overweight and obesity were defined according to age- and gender-specific BMI cut-off points. Multivariate logistic regression analysis was performed to control potential confounders. RESULTS: The prevalence of overweight and obesity was higher among children with 'borderline' or 'abnormal' peer relationship problems compared to 'normal' children. The association of 'abnormal' peer relationship problems was still significant in the final logistic regression model for girls [odds ratio (OR) for overweight 2.0; 95% confidence interval (CI): 1.4-3.0; OR for obesity 2.6; 95% CI: 1.3-5.0]. Among boys the adjusted odds ratio were lower and no longer significant. CONCLUSION: The significantly increased prevalence of overweight and obesity among preschool children with peer relationship problems could not be explained by confounding. It seems evident that there is a relevant co-morbidity of peer relationship problems and obesity in pre-school children pointing to the need of interventions focusing on both physical as well as psychosocial health. PMID- 19681767 TI - n-3 Fatty acid supplementation in burned paediatric patients. AB - AIM: To determine the effect of dietary supplementation with n-3 fatty acids (FA) in paediatric burned patients who had less than 20% of total body surface affected. METHODS: Burned patients were randomly assigned into two groups, one of them received a supplement of n-3 FA during 5 weeks; the other group was considered as not n-3 supplemented burned group. A third group of no burned patients was selected as control. Blood samples were collected at admission and in burned groups at the final of the study. Plasma and erythrocyte phospholipid FA composition and some biochemical parameters related to the clinical evolution: total plasma proteins and C3 and C4 complement proteins were determined. RESULTS: In the early post-burn patients, there is an increase in saturated and monounsaturated FAs in plasma phospholipids, and a decrease in polyunsaturated FAs compared with control. These alterations are in favour of proinflammatory response to burn injury. In n-3 FA supplemented group, these changes were further reverted, and a favourable response in the amount of total plasma proteins and in C3 and C4 proteins of the complement system was demonstrated. CONCLUSION: Dietary n-3 FA supplementation might be beneficial for patients suffering thermal injury. PMID- 19681768 TI - Training response of mitochondrial transcription factors in human skeletal muscle. AB - AIM: Mitochondrial function is essential for physical performance and health. Aerobic fitness is positively associated with mitochondrial (mt) biogenesis in muscle cells through partly unknown regulatory mechanisms. The present study aimed to investigate the influence of exercise and training status on key mt transcription factors in relation to oxidative capacity in human skeletal muscle. METHODS: The basal mRNA and protein levels of mitochondrial transcription factor A (TFAM), mitochondrial transcription factors B1 (TFB1M) or B2 (TFB2M), and mRNA levels of mitochondrial transcription termination factor (mTERF), were measured in a cross-sectional study with elite athletes (EA) and moderately active (MA) and the basal mRNA levels of these factors were measured during a 10-day endurance training programme with (R-leg) and without (NR-leg) restricted blood flow to the working leg. RESULTS: TFAM protein expression was significantly higher in the EA than in the MA, while protein levels of TFB1M and TFB2M were not different between the groups. There was no difference between EA and MA, or any effect with training on TFAM mRNA levels. However, the mRNA levels of TFB1M, TFB2M and mTERF were higher in EA compared with MA. For TFB1M and TFB2M, the mRNA expression was increased in the R-leg after 10 days of training, but not in the NR-leg. mTERF mRNA levels were higher in EA compared with MA. CONCLUSION: This study further establishes that TFAM protein levels are higher in conditions with enhanced oxidative capacity. The mRNA levels of TFB1M and TFB2M are influenced by endurance training, possibly suggesting a role for these factors in the regulation of exercise-induced mitochondrial biogenesis. PMID- 19681769 TI - Acute exercise reverses TRB3 expression in the skeletal muscle and ameliorates whole body insulin sensitivity in diabetic mice. AB - AIM: TRB3 became of major interest in diabetes research when it was shown to interact with and inhibit the activity of Akt. Conversely, physical exercise has been linked to improved glucose homeostasis. Thus, the current study was designed to investigate the effects of acute exercise on TRB3 expression and whole body insulin sensitivity in obese diabetic mice. METHODS: Male leptin-deficient (ob/ob) mice swam for two 3-h-long bouts, separated by a 45-min rest period. After the second bout of exercise, food was withdrawn 6 h before antibody analysis. Eight hours after the exercise protocol, the mice were submitted to an insulin tolerance test (ITT). Gastrocnemius muscle samples were evaluated for insulin receptor (IR) and IRS-1 tyrosine phosphorylation, Akt serine phosphorylation, TRB3/Akt association and membrane GLUT4 expression. RESULTS: Western blot analysis showed that TRB3 expression was reduced in the gastrocnemius of leptin-deficient (ob/ob) mice submitted to exercise when compared with respective ob/ob mice at rest. In parallel, there was an increase in the insulin-signalling pathway in skeletal muscle from leptin-deficient mice after exercise. Furthermore, the GLUT4 membrane expression was increased in the muscle after the exercise protocol. Finally, a single session of exercise improved the glucose disappearance (K(ITT)) rate in ob/ob mice. CONCLUSION: Our results demonstrate that acute exercise reverses TRB3 expression and insulin signalling restoration in muscle. Thus, these results provide new insights into the mechanism by which physical activity ameliorates whole body insulin sensitivity in type 2 diabetes. PMID- 19681770 TI - Moroccan national study on prevalence of mental disorders: a community-based epidemiological study. AB - OBJECTIVE: The main objective of this study was to assess the prevalence of common mental disorders in the Moroccan general population. METHOD: On a systematic representative randomized sample, the Moroccan Arabic version of the Mini International Neuropsychiatric Interview (MINI) was used to assess the prevalence of mood, anxiety, substance, and alcohol abuse disorders. RESULTS: Among 5498 subjects interviewed, 40.1% had at least one current mental disorder. Current major depressive disorder was the most common (26.5%), and at least one anxiety disorder was found in 37% of the sample. Mental disorders were more frequent among female, urban, divorced, and unemployed subjects. CONCLUSION: Mental disorders are common in the Moroccan general population, particularly mood and anxiety disorders. PMID- 19681771 TI - Chronic pain after cardiac surgery: a prospective study. AB - BACKGROUND: Chronic pain is a complication of several surgical procedures. The prevalence of chronic pain reported after cardiac surgery varies from 18% to 61%. However, most studies are retrospective, do not use validated instruments for pain measurement or include only pain at the sternum site. The aim of the present study was to assess chronic pain and health-related quality of life (HRQOL) after cardiac surgery. METHODS: In a prospective, population-based study, we included 534 patients (413 males) and assessed chronic pain and HRQOL before, 6 months after, and 12 months after cardiac surgery. Pain was measured by the Brief Pain Inventory, while HRQOL was measured by the Short-Form 36 (SF-36). RESULTS: Five hundred and twenty-one patients were alive 12 months after surgery; 462 (89%) and 465 (89%) responded after 6 and 12 months, respectively. Chronic pain was reported by 11% of the patients at both measurements. Younger age was associated with chronic pain [odds ratio 0.7 (95% confidence interval: 0.5-0.9)] at 12 months. Patients with chronic pain reported lower scores on seven of eight SF-36 subscales. DISCUSSION: In conclusion, we observed a lower prevalence of chronic pain after cardiac surgery than in previous studies. Still, more than one out of 10 patients reported chronic pain after cardiac surgery. Chronic pain appears to affect HRQOL. Thus, given the large number of patients subjected to cardiac surgery, this study confirms that chronic pain after cardiac surgery is an important health care issue. PMID- 19681772 TI - Sparing the larynx during gynecological laparoscopy: a randomized trial comparing the LMA Supreme and the ETT. AB - BACKGROUND: We designed a prospective randomized single-blind study to compare efficiency and post-operative upper airway morbidity when the laryngeal mask airway (LMA) Supreme is used as an alternative to the endotracheal tube (ETT). METHODS: One hundred and thirty-eight elective pelvic laparoscopic ASA I-II female patients were assigned to receive either the LMA Supreme or the ETT for airway management. Balanced anesthesia and ventilation techniques were standardized to control end-tidal CO(2) and BIS value in the range 4.5-5 kPa and 40-50, respectively, and to maintain adequate hemodynamic stability. A single surgeon blinded to the airway management technique performed all surgical procedures. The ventilation efficiency of each airway was evaluated. Anesthesia- and surgery-related times were calculated and anesthesia details were recorded. Post-operative pain and pharyngolaryngeal morbidity were measured in a blind fashion using a numerical rating scale (NRS) (0-100). RESULTS: Surgery duration was similar in both groups. Airway management duration was shorter with the LMA Supreme. Post-operative pharyngolaryngeal morbidity incidence and all symptoms' intensity were significantly increased after ETT as compared with LMA Supreme anesthesia. At the end of the PACU stage, the incidence and mean NRS of post operative hoarseness were reduced when LMA Supreme was used as an alternative to the ETT (16% vs. 47%; P<0.01 and 9 vs. 19, P<0.01, respectively). CONCLUSION: We demonstrated that choosing an LMA Supreme was an efficient pharyngolaryngeal morbidity-sparing strategy. Moreover, we showed that the LMA Supreme and the ETT were equally effective airways for a routine gynecological laparoscopy procedure. PMID- 19681773 TI - The GlideScope Ranger video laryngoscope can be useful in airway management of entrapped patients. AB - BACKGROUND: Airway management of entrapped patients is challenging and alternatives to endotracheal intubation with a Macintosh laryngoscope must be considered. In this study, the GlideScope Ranger video laryngoscope has been evaluated as an alternative to standard laryngoscopy. METHODS: Eight anaesthesiologists from a Helicopter Emergency Medical Service intubated the trachea of a Laerdal SimMan manikin using the studied laryngoscopes in two scenarios: (A) unrestricted access to the manikin in an ambulance and (B) no access from the head end, simulating an entrapped patient. The time used to secure the airway and the scored level of difficulty were the main variables. RESULTS: In scenario A, all anaesthesiologists managed to secure the airway using both techniques within the 60-s time limit. In scenario B, all secured the airway when using the video laryngoscope, while 50% succeeded with endotracheal intubation using the Macintosh laryngoscope. The difference in the success rate was statistically significant (P=0.025). There were no significant differences in the time spent on endotracheal intubation in the two scenarios or between the devices. All stated that the availability of a video laryngoscope would make drug facilitated intubation a realistic alternative when access to patients is limited. The lack of visual control when using the Macintosh laryngoscope excludes this technique in real-life settings. CONCLUSION: This study suggests that the GlideScope Ranger may be merited in situations requiring endotracheal intubation by an experienced intubator in patient entrapment. Further studies are required to clarify whether performance in patients mimics that in a manikin. PMID- 19681774 TI - Single vs. double stimulation during a lateral sagittal infraclavicular block. AB - BACKGROUND: The objective of this study was to evaluate the influence of single vs. dual control during an ultrasound-guided lateral sagittal infraclavicular block on the efficacy of sensory block and the time of block onset. METHODS: In a prospective manner, 60 adult patients scheduled for distal upper limb surgery were randomly allocated to single (Group S) or double stimulation (Group D) groups. A local anesthetic (LA) mixture of 20 ml of levobupivacaine 5 mg/ml and 20 ml of lidocaine 20 mg/ml with 5 microg/ml epinephrine (total 40 ml) was administered in both groups. In the Group S following a median, an ulnar or a radial nerve response, the entire LA was administered at a single site. In Group D 10 ml of LA was administered following the electrolocation of the musculocutaneous nerve and 30 ml LA was injected following median, ulnar or radial nerves. A successful block was defined as analgesia or anesthesia of all five nerves distal to the elbow. Sensory and motor blocks were tested at 5-min intervals for 30 min. RESULTS: The block was successful in 27 patients in Group S and 28 patients in Group D. The time from starting the block until satisfactory anesthesia was significantly shorter in Group D than in Group S (19.3 vs. 23.2 min) (P<0.05). Total sensory scores were significantly higher in the double stimulation group at 20 and 30 min after the block performance (P<0.05). CONCLUSIONS: Although the block performance time was longer in the double stimulation group, block onset time and extent of anesthesia were more favorable in the double stimulation group. PMID- 19681775 TI - Glutamine administration in patients undergoing cardiac surgery and the influence on blood glutathione levels. AB - BACKGROUND: Cardiac surgery with an extracorporeal circulation cardiopulmonary bypass (CPB) is characterized by an oxidative stress response. Glutathione (GSH) belongs to the major antioxidative defense. In metabolic stress, glutamine (GLN) may be the rate-limiting factor of GSH synthesis. Decreased GLN plasma levels were observed after various critical states. We evaluated, in patients undergoing open heart surgery with CPB, the effects of a peri-operative GLN supplementation on GSH in whole blood and assessed their influence on the Sequential Organ Failure Assessment score and the intensive care unit length of stay. METHODS: In this prospective, randomized, double-blinded study, we included 60 patients (age older than 70 years, ejection fraction <40% or mitral valve replacement) undergoing an elective cardiac surgery with CPB. We randomly assigned each subject to receive an infusion with either GLN (0.5 g/kg/day, group 1) or an isonitrogeneous, isocaloric, isovolemic amino acids solution (group 2) or saline (group 3). RESULTS: From the first post-operative day GLN plasma levels in group 1 were significantly increased compared with the other groups. With saline GSH the levels decreased significantly post-operatively compared with GLN. We observed a significant correlation between GLN delivery and GSH levels. CONCLUSIONS: A peri-operative high-dose GLN infusion increased plasma GLN concentrations and maintained the GSH levels after cardiac surgery with CPB. PMID- 19681776 TI - Life-threatening systemic toxicity and airway compromise from a common European adder bite to the tongue. AB - A 24-year-old man was bit on the tongue by a European common adder. Within 15 min following envenomation, he experienced tongue swelling, hypotension and impaired consciousness. Antihistamine, corticosteroid and crystalloids were administered. Within 105 min of envenomation, increasing oral, pharyngeal and facial oedema compromised the airway, leading to respiratory failure, concomitant with circulatory failure related to hypoxaemia and systemic toxic effects. Acute tracheotomy secured the airway, and two doses of antivenom successfully treated the systemic, toxic effects. The reaction was severe due to rapid and suspected high-dose uptake of venom, underlining the need for early advanced symptomatic treatment with airway control and early and eventually repeated dosing of antivenom. PMID- 19681777 TI - Effect of P6 acustimulation on post-operative nausea and vomiting in patients undergoing a laparoscopic cholecystectomy. AB - BACKGROUND: Non-pharmacologic techniques such as electrical acustimulation may mitigate post-operative nausea and vomiting (PONV). The primary purpose of this study was to investigate the effectiveness of acustimulation on attenuating PONV. Moreover, we tested whether a pre- or a post-induction application of acustimulation results in differences in PONV reduction. METHODS: In this prospective, double-blind, randomized, controlled trial, we studied 200 patients undergoing a laparoscopic cholecystectomy during propofol (induction) fentanyl/isoflurane/atracurium (maintenance) anaesthesia. In the acustimulation group (n=101), subdivided into groups with pre-induction (n=57) and post induction (n=44) acustimulation, an active ReliefBand device was placed at the P6 acupoint. In the sham group (n=99), also subdivided into pre-induction (n=55) or post-induction (n=44) groups, an inactive device was applied instead. The ReliefBand remained in place for 24 h after surgery. Nausea and vomiting/retching were recorded at 2, 6, and 24 h post-operatively. RESULTS: The incidence of early nausea (up to 2 h) was significantly lower in the acustimulation than in the sham group (29% vs. 42%; P=0.043). No significant effect could be detected for retching/vomiting. Moreover, acustimulation showed no effect on PONV after 6 and 24 h. Risk factor analysis (female gender, non-smoker, history of PONV/motion sickness, and post-operative morphine usage) revealed a relative reduction in risk of 40% for nausea (P=0.021) and 55% for retching/vomiting (P=0.048) in patients with three or four risk factors present. The timing of (pre- vs. post induction) acustimulation had no significant effect on PONV reduction. CONCLUSION: Acustimulation at the P6 acupoint reduces early nausea, but not vomiting, after laparoscopic cholecystectomy, irrespective of its pre- or post induction application. PMID- 19681778 TI - The effects of obstructive jaundice on the pharmacodynamics of propofol: does the sensitivity of intravenous anesthetics change among icteric patients? AB - BACKGROUND: Some studies suggest that certain clinical symptoms of cholestasis, such as fatigue and pruritus, result from altered neurotransmission. Patients with obstructive jaundice also have labile blood pressure and heart rate. In the present study, the authors investigated whether obstructive jaundice affects a patient's sensitivity to hypnotics and the haemodynamic profile of propofol. METHODS: Thirty-six ASA physical status I/II/III patients with serum total bilirubin (TBL) from 7.8 to 362.7 micromol/l scheduled for bile duct surgery were recruited. A computer-controlled propofol infusion programmed for effect site target was used to rapidly attain and maintain sequential increase of the compartment concentration (from 1 to 3 microg/ml). Each target-controlled concentration was maintained for about 12 min, and arterial blood samples were drawn for propofol concentration determination. The bispectral index (BIS) and mean arterial pressures (MAP) were used as indices of the propofol effect. The relation between the concentration and the effects was described by the Hill equation. The pharmacodynamic parameters were optimized using a nonlinear mixed effect model. RESULTS: TBL was not a significant covariate of EC(50) for the pharmacodynamic model. For BIS and MAP, the parameters of the pharmacodynamic model were E(max)=75.77%, EC(50)=2.34 microg/ml, and gamma=1.82, and E(max)=47.83%, EC(50)=1.49 microg/ml, and gamma=1.88, respectively. CONCLUSIONS: We demonstrated that obstructive jaundice with serum TBL from 7.8 to 362.7 micromol/l had no effect on propofol pharmacodynamics observed by BIS and MAP. PMID- 19681779 TI - Measurements of functional residual capacity during intensive care treatment: the technical aspects and its possible clinical applications. AB - Direct measurement of lung volume, i.e. functional residual capacity (FRC) has been recommended for monitoring during mechanical ventilation. Mostly due to technical reasons, FRC measurements have not become a routine monitoring tool, but promising techniques have been presented. We performed a literature search of studies with the key words 'functional residual capacity' or 'end expiratory lung volume' and summarize the physiology and patho-physiology of FRC measurements in ventilated patients, describe the existing techniques for bedside measurement, and provide an overview of the clinical questions that can be addressed using an FRC assessment. The wash-in or wash-out of a tracer gas in a multiple breath maneuver seems to be best applicable at bedside, and promising techniques for nitrogen or oxygen wash-in/wash-out with reasonable accuracy and repeatability have been presented. Studies in ventilated patients demonstrate that FRC can easily be measured at bedside during various clinical settings, including positive end-expiratory pressure optimization, endotracheal suctioning, prone position, and the weaning from mechanical ventilation. Alveolar derecruitment can easily be monitored and improvements of FRC without changes of the ventilatory setting could indicate alveolar recruitment. FRC seems to be insensitive to over inflation of already inflated alveoli. Growing evidence suggests that FRC measurements, in combination with other parameters such as arterial oxygenation and respiratory compliance, could provide important information on the pulmonary situation in critically ill patients. Further studies are needed to define the exact role of FRC in monitoring and perhaps guiding mechanical ventilation. PMID- 19681780 TI - Active cooling in traumatic brain-injured patients: a questionable therapy? AB - Hypothermia is shown to be beneficial for the outcome after a transient global brain ischaemia through its neuroprotective effect. Whether this is also the case after focal ischaemia, such as following a severe traumatic brain injury (TBI), has been investigated in numerous studies, some of which have shown a tendency towards an improved outcome, whereas others have not been able to demonstrate any beneficial effect. A Cochrane report concluded that the majority of the trials that have already been published have been of low quality, with unclear allocation concealment. If only high-quality trials are considered, TBI patients treated with active cooling were more likely to die, a conclusion supported by a recent high-quality Canadian trial on children. Still, there is a belief that a modified protocol with a shorter time from the accident to the start of active cooling, longer cooling and rewarming time and better control of blood pressure and intracranial pressure would be beneficial for TBI patients. This belief has led to the instigation of new trials in adults and in children, including these types of protocol adjustments. The present review provides a short summary of our present knowledge of the use of active cooling in TBI patients, and presents some tentative explanations as to why active cooling has not been shown to be effective for outcome after TBI. We focus particularly on the compromised circulation of the penumbra zone, which may be further reduced by the stress caused by the difference in thermostat and body temperature and by the hypothermia-induced more frequent use of vasoconstrictors, and by the increased risk of contusional bleedings under hypothermia. We suggest that high fever should be reduced pharmacologically. PMID- 19681781 TI - Improved outcome after trauma care in university-level intensive care units. AB - BACKGROUND: Centralized trauma care has been shown to be associated with improved patient outcome. We compared the outcomes of trauma patients in relation to the size of the intensive care unit (ICU) using a large Finnish database. METHODS: A national prospectively collected ICU data registry was used for analysis. All adult trauma admissions excluding isolated head trauma and burns registered from July 1999 to December 2006 were analyzed. Data from 22 ICUs were available. The non-university-affiliated units were categorized according to the number of beds and referral population as small, mid size and large. Acute physiology and chronic health evaluation (APACHE II)- and sequential organ failure assessment (SOFA)-adjusted mortalities were compared between the units. RESULTS: There were 2067 trauma admissions that fulfilled the inclusion criteria; 38% were treated in the university hospitals, 26% in large non-teaching ICUs, 20% in mid size ICUs and 15% in small ICUs. The crude hospital mortality was 5.6%, being 4.7% in university ICU and 6.6% in mid size ICU. In two subgroup analyses of severely ill trauma patients with APACHE II points >25 or SOFA score >8 points, respectively, hospital mortality was significantly lower in university ICUs. CONCLUSIONS: University-level hospitals were associated with better outcomes with critically ill trauma patients. These results can be used in planning future organization of trauma patient care in Finland. PMID- 19681782 TI - Levosimendan cardioprotection in acutely beta-1 adrenergic receptor blocked open chest pigs. AB - BACKGROUND: Levosimendan and volatile anesthetics have myocardial pre conditioning effects. beta-1 adrenergic receptor antagonists may inhibit the protective effect of volatile anesthetics. No information exists as to whether this also applies to the pre-conditioning effect of levosimendan. We therefore investigated whether levosimendan added to metoprolol would demonstrate a cardioprotective effect. METHODS: Three groups of anesthetized open chest pigs underwent 30 min of myocardial ischemia and 90 min of reperfusion by temporary occlusion of the largest side branch from the circumflex artery or the left anterior descending artery. One group (CTRL) served as a control, in another group (BETA), a metoprolol-loading dose was intravenously injected 30 min before ischemia, and in a third group (BETA+L), a levosimendan infusion was added to metoprolol. Myocardial tissue concentrations of glucose, glycerol, and lactate/pyruvate ratio as the primary end-points were investigated with microdialysis in ischemic and non-ischemic tissues. RESULTS: At the end of the ischemic period, statistically significant differences were only found between CTRL and BETA+L in the ischemic myocardium, with a lower lactate/pyruvate ratio, lower glycerol, and higher glucose concentrations in BETA+L as compared with CTRL. There were no differences in non-ischemic myocardium. From 10 to 90 min of reperfusion, no more differences were found between groups. CONCLUSION: The cardioprotective effect of levosimendan on ischemic metabolism with a reduction in the myocardial lactate/pyruvate ratio, less glycerol accumulation, and better preserved glucose concentration does not seem to be prevented by beta-1 adrenergic receptor antagonism with metoprolol. PMID- 19681783 TI - Visualization of alveolar recruitment in a porcine model of unilateral lung lavage using 3He-MRI. AB - BACKGROUND: In the acute respiratory distress syndrome potentially recruitable lung volume is currently discussed. (3)He-magnetic resonance imaging ((3)He-MRI) offers the possibility to visualize alveolar recruitment directly. METHODS: With the approval of the state animal care committee, unilateral lung damage was induced in seven anesthetized pigs by saline lavage of the right lungs. The left lung served as an intraindividual control (healthy lung). Unilateral lung damage was confirmed by conventional proton MRI and spiral-CT scanning. The total aerated lung volume was determined both at a positive end-expiratory pressure (PEEP) of 0 and 10 mbar from three-dimensionally reconstructed (3)He images, both for healthy and damaged lungs. The fractional increase of aerated volume in damaged and healthy lungs, followed by a PEEP increase from 0 to 10 mbar, was compared. RESULTS: Aerated gas space was visualized with a high spatial resolution in the three-dimensionally reconstructed (3)He-MR images, and aeration defects in the lavaged lung matched the regional distribution of atelectasis in proton MRI. After recruitment and PEEP increase, the aerated volume increased significantly both in healthy lungs from 415 ml [270-445] (median [min-max]) to 481 ml [347-523] and in lavaged lungs from 264 ml [71-424] to 424 ml [129-520]. The fractional increase in lavaged lungs was significantly larger than that in healthy lungs (healthy: 17% [11-38] vs. lavage: 42% [14-90] (P=0.031). CONCLUSION: The (3)He-MRI signal might offer an experimental approach to discriminate atelectatic vs. poor aerated lung areas in a lung damage animal model. Our results confirm the presence of potential recruitable lung volume by either alveolar collapse or alveolar flooding, in accordance with previous reports by computed tomography. PMID- 19681784 TI - Late sensory function after intraoperative capsaicin wound instillation. AB - BACKGROUND: Intense capsaicin-induced C-fiber stimulation results in reversible lysis of the nerve soma, thereby making capsaicin wound instillation of potential interest for the treatment of post-operative pain. Clinical histological and short-term sensory studies suggest that the C-fiber function is partly re established after skin injection of capsaicin. However, no study has evaluated the long-term effects of wound instillation of purified capsaicin on sensory functions. METHODS: Patients included in a double-blind placebo-controlled randomized study of the analgesic effect of capsaicin after groin hernia repair were examined by quantitative sensory testing before, 1 week and 2 years post operatively. The primary endpoint was occurrence of hyperalgesia/allodynia. The secondary endpoints were acute and late sensory changes between the two patient groups. Patients were blinded to the allocated treatment. RESULTS: Twenty (100%) capsaicin and 16 (76%) placebo-treated patients were seen at the 2 1/2 year follow-up. Hyperalgesia was seen in five capsaicin- vs. one placebo-treated patient (P=0.2). The mechanical detection threshold was significantly increased on the operated side in the capsaicin vs. placebo group at the 1-week follow-up (P<0.05), but was not different at the 2 1/2 year follow-up (P=0.3). There were no other significant differences in sensory function on the operated side between groups at the pre-operative, 1-week or 2 1/2 year post-operative follow-up (P>0.05). The sensory function on the contralateral side was comparable between groups throughout the study (P>0.1). CONCLUSION: This small-volume study calls for further long-term safety studies of wound capsaicin instillation. PMID- 19681785 TI - Brain death and its implications for management of the potential organ donor. AB - The systemic physiologic changes that occur during and after brain death affect all organs suitable for transplantation. Major changes occur in the cardiovascular, pulmonary, endocrine, and immunological systems, and, if untreated may soon result in cardiovascular collapse and somatic death. Understanding these complex physiologic changes is mandatory for developing effective strategies for donor resuscitation and management in such a way that the functional integrity of potentially transplantable organs is maintained. This review elucidates these physiological changes and their consequences, and based on these consequences the rationale behind current medical management of brain dead organ donors is discussed. PMID- 19681787 TI - Colchicine and ocular surface changes in familial Mediterranean fever. PMID- 19681786 TI - Automated pre-ejection period variation predicts fluid responsiveness in low tidal volume ventilated pigs. AB - INTRODUCTION: The respiratory variation in the pre-ejection period (Delta PEP) has been used to predict fluid responsiveness in mechanically ventilated patients. Recently, we automated this parameter and indexed it to tidal volume (PEPV) and showed that it was a reliable predictor for post-cardiac surgery, mainly paced, patients ventilated with low tidal volumes. The aims of the present animal study were to investigate PEPV's ability to predict fluid responsiveness under different fluid loading conditions and natural heart rates during low tidal volume ventilation (6 ml/kg) and to compare the performance of PEPV with other markers of fluid responsiveness. METHODS: Eight prone, anesthetized piglets (23 27 kg) ventilated with tidal volumes of 6 ml/kg were subjected to a sequence of 25% hypovolemia, normovolemia, and 25% and 50% hypervolemia. PEPV, Delta PEP, pulse pressure variation (PPV), central venous pressure (CVP), and pulmonary artery occlusion pressure (PAOP) were measured before each volume expansion. RESULTS: Sensitivity was 89% and specificity was 93% for PEPV, 78% and 93% for Delta PEP, 89% and 100% for PPV, 78% and 93% for CVP, and 89% and 87% for PAOP. CONCLUSION: PEPV predicts fluid responsiveness in low tidal volume ventilated piglets. PMID- 19681788 TI - Intravitreal bevacizumab for retinal capillary haemangioma: longterm results. PMID- 19681789 TI - Kimura's disease in the medial canthus. PMID- 19681790 TI - Management of congenital nasolacrimal duct obstruction. AB - Our review aims to provide an update of management protocols for congenital nasolacrimal duct obstruction (CNDO). Although early probing performed before the age of 1 year was traditionally recommended, many reports have since confirmed high frequencies of spontaneous resolution during the first year of life. Accordingly, a 'wait-and-see' approach, combined with conservative therapies, is judged to be the best option in infants aged<1 year. By contrast, persistent obstruction beyond 1 year of age warrants probing as a first-line interventional therapy. However, the optimal timing for probing remains controversial. Although there remains a high possibility of spontaneous resolution after the first year of age, this must be balanced against the decrease in success rates for probing that accompanies advancing age. If conservative management fails, persistent CNDO beyond 1 year of age should be managed either by further observation or by primary probing according to the severity of symptoms. In patients in whom probing fails, advanced treatment such as balloon catheter dilation, silicone tube intubation or dacryocystorhinostomy may be considered. PMID- 19681791 TI - Diagnostic accuracy of Heidelberg Retina Tomograph III classifications in a Turkish primary open-angle glaucoma population. AB - PURPOSE: This study aimed to evaluate the diagnostic accuracy of Moorfields regression analysis (MRA) and the glaucoma probability score (GPS) in primary open-angle glaucoma (POAG) and to measure the level of agreement between the two algorithms in classifying eyes as normal or abnormal in a Turkish population. METHODS: We prospectively selected 184 healthy subjects and 158 subjects with POAG, who underwent an ophthalmological examination, visual field analysis and imaging with the Heidelberg Retina Tomograph II, using HRT III software, Version 3.0. The diagnostic accuracies of the two classifications were measured when the borderline was taken as either normal (highest specificity criteria) or abnormal (highest sensitivity criteria). The agreement between them was calculated using the unweighted kappa (kappa) coefficient. RESULTS: Optic nerve head topographic parameters showed statistically significant differences between the control and POAG groups (p < 0.001). The parameters with the highest area under the receiver operating characteristic curves were global GPS (0.86), cup : disc area (0.85), rim : disc area (0.85) and vertical cup : disc (0.85). According to the highest specificity criteria, MRA had a sensitivity of 67.7% and a specificity of 95.1%, whereas the GPS had a sensitivity of 70.9% and a specificity of 88.0%. According to the highest sensitivity criteria, MRA had a sensitivity of 81.0% and a specificity of 75.0%, whereas the GPS had a sensitivity of 89.2% and a specificity of 57.6%. A moderate agreement of 68% (233 eyes) with a kappa coefficient of 0.51 was found between MRA and the GPS. CONCLUSIONS: The GPS automated classification showed similar sensitivity to MRA, but considerably lower specificity, when applied in a Turkish population. PMID- 19681792 TI - Intravitreal methotrexate in therapeutically resistant exudative age-related macular degeneration. PMID- 19681793 TI - Retinal arterioles have impaired reactivity to hyperoxia in type 1 diabetes. AB - PURPOSE: Diabetes has adverse effects on the retinal microvasculature. The purpose of this study was to compare the effects of inhalation of hypoxic, hyperoxic and normoxic-hypercapnic gas mixtures on retinal vessel diameter in people with and without diabetes. METHODS: Sixty-one participants (aged 24-50 years) 29 with (male : female ratio 2.6 : 1) and 32 without (male : female ratio 0.7 : 1) diabetes, inhaled hypoxic, hyperoxic and normoxic-hypercapnic gas mixtures for 3-5 mins. The diameters of arterioles and venules were measured using digital retinal images taken before and after gas inhalation. RESULTS: There was no significant difference in the diameters of arterioles and venules prior to gas inhalation in people with and without diabetes. Inhalation of the hyperoxic gas mixture caused a statistically significant decrease in arteriolar and venular diameters without altering mean arterial pressure significantly. Arteriolar vasoconstriction in response to the hyperoxic gas mixture was significantly reduced in people with diabetes (3.95% versus 7.75%; p = 0.04), but venular vasoconstriction did not differ significantly. A hypoxic gas mixture caused increased arteriolar and venular diameter and a normoxic-hypercapnic gas mixture had no significant effect on vessel diameter. Responses to hypoxic and normoxic-hypercapnic gas did not differ significantly between diabetes and non diabetes subjects. CONCLUSIONS: Type 1 diabetes impairs retinal arteriolar responses to hyperoxia. Abnormalities in retinal arteriolar reactivity in response to oxygen may play a role in the development of diabetic retinopathy and this technique may represent a simple means of identifying early abnormalities in the reactivity of retinal arterioles in diabetes. PMID- 19681794 TI - New insight into the pathology of macular detachment associated with an optic disc pit. PMID- 19681795 TI - Bridging the gap between adult and paediatric outcomes in HIV-1 vertically infected children: a single-centre comparison with adult data. AB - Prognosis of HIV-1 infection dramatically improved during the last decade. Meanwhile, treatment-induced virological success has always been different in adult and children patients. AIM: To compare 10 years of follow up in HIV-1 vertically infected children and adult patients. METHODS: Monocentric retrospective longitudinal analysis of vertically HIV-1-infected children and adult patients followed in the Nice University Hospital between 1999 and 2008. Immunological, virological and antiretroviral treatment data were recorded. RESULTS: Forty children and 1752 adult patients were included. Between 1996 and 2008, the percentage of children receiving HAART increased from 3.2% to 91%. Mean CD4% in the paediatric group remained stable between 29 +/- 8.1% in 1998 and 30 +/- 9.4% in 2008. Mean adult CD4-cell count significantly increased from 410 in 1998 to 556 cells/mL in 2008. Logistic regression analysis showed that the children-to-adult difference for indetectability (HIV PCR-RNA below 400 copies/mL) was significant (p < 0.0001) with an odds ratio of 0.61 (CI(95th): 0.52-0.72). Year-to-patient interaction was also significant with a decreasing divergence over time (p: 0.038). CONCLUSION: Nowadays as in adult patients, the control of HIV-1 replication is achieved in nearly eight of 10 children and the percentage of patients with severe immunodeficiency dramatically decreased compared with the mid 1990s. PMID- 19681797 TI - Hypothalamic-pituitary-adrenal axis and smoking and drinking onset among adolescents: the longitudinal cohort TRacking Adolescents' Individual Lives Survey (TRAILS). AB - AIMS: We examined within a prospective longitudinal study whether cortisol levels were associated with smoking or drinking behaviours, taking parental substance use into account. DESIGN: The influence of parental substance use on cortisol levels of their adolescent offspring at age 10-12 years was examined. Next, cortisol levels of adolescents who initiated smoking or drinking at the first data collection (age 10-12) were compared to non-users. Finally, we examined whether cortisol levels could predict new onset and frequency of smoking and drinking 2 years later. SETTING AND PARTICIPANTS: First and second assessment data of the TRacking Adolescents' Individual Lives Survey (TRAILS) were used, including 1768 Dutch adolescents aged 10-12 years, who were followed-up across a period of 2 years. MEASUREMENTS: Cortisol was measured in saliva samples at awakening, 30 minutes later, and at 8 p.m. at age 10-12. Self-reported substance use at age 10-12 and 13-14, and parental self-reported substance use were used. FINDINGS: Only maternal substance use was related to slightly lower adolescent cortisol levels at 8 p.m. Both maternal and paternal substance use were associated with adolescent smoking and drinking at age 13-14, although fathers' use only predicted the amount used and not the chance of ever use. Finally, higher cortisol levels were related moderately to current smoking and future frequency of smoking, but not to alcohol use. CONCLUSIONS: In a general population, parental heavy substance use does not seem to affect cortisol levels consistently in their offspring. We found some evidence for higher, instead of lower, hypothalamic-pituitary-adrenal axis activity as a predictor of smoking in early adolescence. PMID- 19681796 TI - Overload-induced skeletal muscle extracellular matrix remodelling and myofibre growth in mice lacking IL-6. AB - AIM: Overloading healthy skeletal muscle produces myofibre hypertrophy and extracellular matrix remodelling, and these processes are thought to be interdependent for producing muscle growth. Inflammatory cytokine interleukin-6 (IL-6) gene expression is induced in overloaded skeletal muscle, and the loss of this IL-6 induction can attenuate the hypertrophic response to overload (OV). Although the OV induction of IL-6 in skeletal muscle may be an important regulator of inflammatory processes and satellite cell proliferation, less is known about its role in the regulation of extracellular matrix remodelling. The purpose of the current study was to examine if OV-induced extracellular matrix remodelling, muscle growth, and associated gene expression were altered in mice that lack IL-6, when compared with wild-type mice. METHODS: Male C57/BL6 (WT) and C57/BL6 x IL-6(-/-) (IL-6(-/-)) mice (10 weeks of age) were assigned to either a sham control or synergist ablation OV treatments for 3, 21 or 56 days. RESULT: Plantaris muscle mass increased 59% in WT and 116% in IL-6(-/-) mice after 21 day OV. Myofibre CSA was also increased by 21 day OV in both WT and IL-6(-/-) mice. OV induced a twofold greater increase in the volume of non-contractile tissue in IL-6(-/-) muscle compared to WT. OV also induced a significantly greater accumulation of hydroxyproline and procollagen-1 mRNA in IL-6(-/-) muscle, when compared with WT muscle after 21 day OV. Transforming growth factor-beta and insulin-like growth factor-1 mRNA expression were also induced to a greater extent in IL-6(-/-) muscle when compared with WT muscle after 21 day OV. There was no effect of IL-6 loss on the induction of myogenin, and cyclin D1 mRNA expression after 3 day OV. However, MyoD mRNA expression in 3 day OV IL-6(-/-) muscle was attenuated when compared with WT OV mice. CONCLUSION: IL-6 appears to be necessary for the normal regulation of extracellular matrix remodelling during OV-induced growth. PMID- 19681798 TI - 'I have no interest in drinking': a cross-national comparison of reasons why men and women abstain from alcohol use. AB - AIMS: To examine country differences in reasons for abstaining including the association of reasons with country abstaining rate and drinking pattern. PARTICIPANTS: Samples of men and women from eight countries participating in the GENACIS (Gender Alcohol and Culture: an International Study) project. METHODS: Surveys were conducted with 3338 life-time abstainers and 3105 former drinkers. Respondents selected all applicable reasons for not drinking from a provided list. Analyses included two-level hierarchical linear modelling (HLM) regression. FINDINGS: Reasons for abstaining differed significantly for life-time abstainers compared to former drinkers, by gender and age, and by country-level abstaining rate and frequency of drinking. Life-time abstainers were more likely than former drinkers to endorse 'no interest', 'religion' and 'upbringing' and more reasons overall. Gender differences, especially among former drinkers, suggested that norms restricting drinking may influence reasons that women abstain ('no interest', 'not liking taste') while drinking experiences may be more important considerations for men ('afraid of alcohol problems', 'bad effect on activities'). Younger age was associated with normative reasons ('no interest', 'taste', 'waste of money') and possibly bad experiences ('afraid of problems'). Reasons such as 'religion', 'waste of money' and 'afraid of alcohol problems' were associated with higher country-level rates of abstaining. Higher endorsement of 'drinking is bad for health' and 'taste' were associated with a country pattern of less frequent drinking while 'not liking effects' was associated with higher drinking frequency. CONCLUSIONS: Reasons for abstaining depend on type of abstainer, gender, age and country drinking norms and patterns. PMID- 19681799 TI - Preventing growth in amphetamine use: long-term effects of the Midwestern Prevention Project (MPP) from early adolescence to early adulthood. AB - AIM: The aim of the current study was to examine the long-term effect of an early adolescent substance abuse prevention program on trajectories and initiation of amphetamine use into early adulthood. DESIGN: Eight middle schools were assigned randomly to a program or control condition. The randomized controlled trial followed participants through 15 waves of data, from ages 11-28 years. This longitudinal study design includes four separate periods of development from early adolescence to early adulthood. SETTING: The intervention took place in middle schools. PARTICIPANTS: A total of 1002 adolescents from one large mid western US city were the participants in the study. INTERVENTION: The intervention was a multi-component community-based program delivered in early adolescence with a primary emphasis on tobacco, alcohol and marijuana use. MEASURES: At each wave of data collection participants completed a self-report survey that included questions about life-time amphetamine use. FINDINGS: Compared to a control group, participants in the Midwestern Prevention Project (MPP) intervention condition had reduced growth (slope) in amphetamine use in emerging adulthood, a lower amphetamine use intercept at the commencement of the early adulthood and delayed amphetamine use initiation. CONCLUSIONS: The pattern of results suggests that the program worked first to prevent amphetamine use, and then to maintain the preventive effect into adulthood. Study findings suggest that early adolescent substance use prevention programs that focus initially on the 'gateway' drugs have utility for long-term prevention of amphetamine use. PMID- 19681800 TI - Interventions with injection drug users in Ukraine. AB - AIMS: To assess the effectiveness of a brief human immunodeficiency virus (HIV) testing and counseling intervention compared to a more time-consuming and expensive street-based intervention with injection drug users (IDUs). DESIGN: Cross-over experimental design in which 900 IDUs were recruited, followed by a 'wash-out' period with no recruitment, a reversal of intervention assignment areas and an additional recruitment of 900 IDUs with baseline and 6-month follow up assessments. SETTING: Kiev, Odessa and Makeevka/Donesk Ukraine. PARTICIPANTS: A total of 1798 IDUs. MEASUREMENTS: HIV testing and audio computer-assisted self interview (ACASI) data on socio-demographics, drug use and injection and sex related risk behaviors. FINDINGS: Participants in both conditions reduced their injection and sex risks significantly; however, there was little difference in outcomes between conditions. IDUs who knew they were HIV-infected at baseline were significantly more likely to practice safe sex than those unaware or HIV negative; those who first learned that they were infected at baseline changed their safe sex practices significantly more than those who already knew that they were infected at baseline and those who were HIV-negative. Younger IDUs and those injecting for a shorter period of time reported higher injection and sex risk behaviors following interventions. CONCLUSIONS: Awareness of HIV infection by street-recruited drug injectors is associated with reduced sex risks. Additional interventions are required for younger IDUs and those injecting for shorter periods of time. PMID- 19681801 TI - Changes in per capita alcohol sales during the partial privatization of British Columbia's retail alcohol monopoly 2003-2008: a multi-level local area analysis. AB - AIM: To investigate the independent effects on liquor sales of an increase in (a) the density of liquor outlets and (b) the proportion of liquor stores in private rather than government ownership in British Columbia between 2003/4 and 2007/8. DESIGN: The British Columbia Liquor Distribution Branch provided data on litres of ethanol sold through different types of outlets in 89 local health areas of the province by beverage type. Multi-level regression models were used to examine the relationship between per capita alcohol sales and outlet densities for different types of liquor outlet after adjusting for potential confounding social, economic and demographic factors as well as spatial and temporal autocorrelation. SETTING: Liquor outlets in 89 local health areas of British Columbia, Canada. FINDINGS: The number of private stores per 10,000 residents was associated significantly and positively with per capita sales of ethanol in beer, coolers, spirits and wine, while the reverse held for government liquor stores. Significant positive effects were also identified for the number of bars and restaurants per head of population. The percentage of liquor stores in private versus government ownership was also associated significantly with per capita alcohol sales when controlling for density of liquor stores and of on-premise outlets (P < 0.01). CONCLUSION: The trend towards privatisation of liquor outlets between 2003/04 and 2007/08 in British Columbia has contributed to increased per capita sales of alcohol and hence possibly also to increased alcohol-related harm. PMID- 19681802 TI - Test-re-test reliability of DSM-IV adopted criteria for 3,4 methylenedioxymethamphetamine (MDMA) abuse and dependence: a cross-national study. AB - AIMS: This study evaluated the prevalence and reliability of DSM-IV adopted criteria for 3,4-methylenedioxymethamphetamine (MDMA) abuse and dependence with a purpose to determine whether it is best conceptualized within the category of hallucinogens, amphetamines or its own category. DESIGN: Test-re-test study. PARTICIPANTS: MDMA users (life-time use >5 times) were recruited in St Louis, Miami and Sydney (n=593). The median life-time MDMA consumption was 50 pills at the baseline. MEASUREMENTS: The computerized Substance Abuse Module for Club Drug (CD-SAM) was used to assess MDMA abuse and dependence. The Discrepancy Interview Protocol (DIP) was used to determine the reasons for the discrepant responses between the two interviews. Reliability of diagnoses, individual diagnostic criteria and withdrawal symptoms was examined using the kappa coefficient (kappa). findings for baseline data, 15% and 59% met MDMA abuse and dependence, respectively. Substantial test-re-test reliability of the diagnoses was observed consistently across cities (kappa=0.69). 'Continued use despite knowledge of physical/psychological problems' (87%) and 'withdrawal' (68%) were the two most prevalent dependence criteria. 'Physically hazardous use' was the most prevalent abuse criterion. Six dependence criteria and all abuse criteria were reported reliably across cities (kappa: 0.53-0.77). Seventeen of 19 withdrawal symptoms showed consistency in the reliability across cities. The most commonly reported reason for discrepant responses was 'interpretation of question changed'. Only a small proportion of the total discrepancies were attributed to lying or social desirability. CONCLUSION: The adopted DSM-IV diagnostic classification for MDMA abuse and dependence was moderately reliable across cities. findings on MDMA withdrawal support the argument that MDMA should be separated from other hallucinogens in DSM. PMID- 19681803 TI - Cigarettes and social differentiation in France: is tobacco use increasingly concentrated among the poor? AB - AIMS: This paper aimed to assess whether the increase of social differentiation of smoking is observed in France. DESIGN AND SETTING: Five cross-sectional telephone surveys conducted in France between 2000 and 2007. PARTICIPANTS: The surveys were conducted among national representative samples of French subjects aged 18-75 years (n=12 256, n=2906, n=27 499, n=2887, n=6007 in 2000, 2003, 2005, 2006 and 2007, respectively). We focused on three groups: executives, manual workers and the unemployed. MEASUREMENTS: Time trends of smoking prevalence were assessed, and socio-economic factors (especially occupation and job status) associated with smoking were identified and compared in 2000 and 2005. We also computed respondents' equivalized household consumption (EHI) and their cigarette budget to assess the financial burden of smoking. FINDINGS: Between 2000 and 2007, smoking prevalence decreased by 22% among executive managers and professionals and by 11% among manual workers, and did not decrease among the unemployed. Indicators of an underprivileged social situation were associated more markedly with smoking in 2005 than in 2000. In addition, the falling-off of smoking initiation occurred later and was less marked among manual workers than it was among executive managers and professionals. Finally, in 2005 15% of French smokers devoted at least 20% of their EHI to the purchase of cigarettes, versus only 5% in 2000, and smoking weighted increasingly heavily on the poorest smokers' budgets. CONCLUSIONS: While these results point out an increased social differentiation in tobacco use, they underline the need to design and implement other forms of action to encourage people to quit, in particular targeting individuals belonging to underprivileged groups. PMID- 19681805 TI - Alcohol and Russian mortality: a continuing crisis. AB - BACKGROUND: Russia remains in the grip of a mortality crisis in which alcohol plays a central role. In 2007, male life expectancy at birth was 61 years, while for females it was 74 years. Alcohol is implicated particularly in deaths among working-age men. AIMS: To review the current state of knowledge about the contribution of alcohol to the continuing very high mortality seen among Russian adults RESULTS: Conservative estimates attribute 31-43% of deaths among working age men to alcohol. This latter estimate would imply a minimum of 170 000 excess deaths due to hazardous alcohol consumption in Russia per year. Men drink appreciably more than women in Russia. Hazardous drinking is most prevalent among people with low levels of education and those who are economically disadvantaged, partly because some of the available sources of ethanol are very cheap and easy to obtain. The best estimates available suggest that per capita consumption among adults is 15-18 litres of pure ethanol per year. However, reliable estimation of the total volume of alcohol consumed per capita in Russia is very difficult because of the diversity of sources of ethanol that are available, for many of which data do not exist. These include both illegal spirits, as well as legal non beverage alcohols (such as medicinal tinctures). In 2006 regulations were introduced aimed at reducing the production and sale of non-beverage alcohols that are commonly drunk. These appear to have been only partially successful. CONCLUSION: There is convincing evidence that alcohol plays an important role in explaining high mortality in Russia, in particular among working age men. However, there remain important uncertainties about the precise scale of the problem and about the health effects of the distinctive pattern of alcohol consumption that is prevalent in Russia today. While there is a need for further research, enough is known to justify the development of a comprehensive inter sectoral alcohol control strategy. The recent fall in life expectancy in Russia should give a renewed urgency to attempts to move the policy agenda forward. PMID- 19681804 TI - Subtypes of major depression in substance dependence. AB - AIMS: This study evaluated features that differentiate subtypes of major depressive episode (MDE) in the context of substance dependence (SD). Design Secondary data analysis using pooled data from family-based and case-control genetic studies of SD. SETTING: Community recruitment through academic medical centers. PARTICIPANTS: A total of 1929 unrelated subjects with alcohol and/or drug dependence. MEASUREMENTS: Demographics, diagnostic criteria for psychiatric and substance use disorders and related clinical features were obtained using the Semi-Structured Assessment for Drug Dependence and Alcoholism. We compared four groups: no life-time MDE (no MDE), independent MDE only (I-MDE), substance induced MDE only (SI-MDE) and both types of MDE. FINDINGS: Psychiatric measures were better predictors of MDE subtype than substance-related or socio-demographic ones. Subjects with both types of MDE reported more life-time depressive symptoms and comorbid anxiety disorders and were more likely to have attempted suicide than subjects with I-MDE or SI-MDE. Subjects with both types of MDE, like those with I-MDE, were also more likely than subjects with SI-MDE to be alcohol dependent only than either drug-dependent only or both alcohol- and drug dependent. CONCLUSIONS: SD individuals with both types of MDE have greater psychiatric severity than those with I-MDE only or SI-MDE only. These and other features that distinguish among the MDE subtypes have important diagnostic and potential therapeutic implications. PMID- 19681806 TI - Unplanned attempts to quit smoking: missed opportunities for health promotion? AB - OBJECTIVES: To investigate the occurrence, determinants and reported success of unplanned and planned attempts to quit smoking, and sources of support used in these attempts. DESIGN: Cross-sectional questionnaire survey of 3512 current and ex-smokers. SETTING: Twenty-four general practices in Nottinghamshire, UK. PARTICIPANTS: Individuals who reported making a quit attempt within the last 6 months. MEASUREMENTS: Occurrence, triggers for, support used and success of planned and unplanned quit attempts. RESULTS: A total of 1805 (51.4%) participants returned completed questionnaires, reporting 394 quit attempts made within the previous 6 months of which 37% were unplanned. Males were significantly more likely to make an unplanned quit attempt [adjusted odds ratio (OR) 1.60, 95% confidence interval (CI) 1.04-2.46], but the occurrence of unplanned quit attempts did not differ significantly by socio-economic group or amount smoked. The most common triggers for unplanned quit attempts were advice from a general practitioner or health professional (27.9%) and health problems (24.5%). 5.4% and 4.1% of unplanned quit attempts used National Health Service cessation services on a one to one and group basis, respectively, and more than half (51.7%) were made without any support. Nevertheless, unplanned attempts were more likely to be reported to be successful (adjusted OR 2.01, 95% CI 1.23-3.27, P < 0.01). CONCLUSIONS: Unplanned quit attempts are common among smokers in all socio-demographic groups, are triggered commonly by advice from a health professional and are more likely to succeed; however, the majority of these unplanned attempts are unsupported. It is important to develop methods of providing behavioural and/or pharmacological support for these attempts, and determine whether these increase cessation rates still further. PMID- 19681807 TI - How confident should we be that smoking cessation treatments work? AB - AIM: To determine (i) the concordance among recent meta-analyses about which treatments for smoking cessation are efficacious; (ii) the similarity of odds ratios (ORs) across meta-analyses; and (iii) among the validated treatments, the proportion of studies that found higher quit rates. METHODS: Computerized literature search for meta-analyses during the last 5 years in PubMed and PsychInfo. Data were extracted from summary tables of overall effect of validated treatments. RESULTS: Fourteen meta-analyses agreed 100% on the presence/absence of efficacy of 17 proven treatments. The ORs differed by <0.5 in 72/76 of the comparisons of meta-analyses. Among 37 comparisons in 33 comparisons, >85% of the studies reported numerical superiority for the active treatment. CONCLUSIONS: The efficacy of treatments for smoking cessation are extremely reliable. This argues for inclusion of treatment as an essential feature of tobacco control and clinical practice and argues for reimbursement of smoking cessation treatments on a par with other medical and behavioral disorders. PMID- 19681808 TI - Stress-induced responses of human skin fibroblasts in vitro reflect human longevity. AB - Unlike various model organisms, cellular responses to stress have not been related to human longevity. We investigated cellular responses to stress in skin fibroblasts that were isolated from young and very old subjects, and from offspring of nonagenarian siblings and their partners, representatives of the general population. Fibroblasts were exposed to rotenone and hyperglycemia and assessed for senescence-associated beta-galactosidase (SA-beta-gal) activity by flow cytometry. Apoptosis/cell death was measured with the Annexin-V/PI assay and cell-cycle analysis (Sub-G1 content) and growth potential was determined by the colony formation assay. Compared with fibroblasts from young subjects, baseline SA-beta-gal activity was higher in fibroblasts from old subjects (P = 0.004) as were stress-induced increases (rotenone: P < 0.001, hyperglycemia: P = 0.027). For measures of apoptosis/cell death, fibroblasts from old subjects showed higher baseline levels (Annexin V+/PI+ cells: P = 0.040, Sub-G1: P = 0.014) and lower stress-induced increases (Sub-G1: P = 0.018) than fibroblasts from young subjects. Numbers and total size of colonies under nonstressed conditions were higher for fibroblasts from young subjects (P = 0.017 and 0.006, respectively). Baseline levels of SA-beta-gal activity and apoptosis/cell death were not different between fibroblasts from offspring and partner. Stress-induced increases were lower for SA-beta-gal activity (rotenone: P = 0.064, hyperglycemia: P < 0.001) and higher for apoptosis/cell death (Annexin V+/PI- cells: P = 0.041, Annexin V+/PI+ cells: P = 0.008). Numbers and total size of colonies under nonstressed conditions were higher for fibroblasts from offspring (P = 0.001 and 0.024, respectively) whereas rotenone-induced decreases were lower (P = 0.008 and 0.004, respectively). These data provide strong support for the hypothesis that in vitro cellular responses to stress reflect the propensity for human longevity. PMID- 19681809 TI - Clinical trial: efficacy and safety of dexlansoprazole MR 60 and 90 mg in healed erosive oesophagitis - maintenance of healing and symptom relief. AB - BACKGROUND: Dexlansoprazole MR, a modified-release formulation of dexlansoprazole, an enantiomer of lansoprazole, effectively heals erosive oesophagitis. AIM: To assess dexlansoprazole MR in maintaining healed erosive oesophagitis. METHODS: Patients (n = 451) with erosive oesophagitis healed in either of two dexlansoprazole MR healing trials randomly received dexlansoprazole MR 60 or 90 mg or placebo once daily in this double-blind trial. The percentage of patients who maintained healing at month 6 was analysed using life table and crude rate methods. Secondary endpoints were percentages of nights and of 24-h days without heartburn based on daily diaries. RESULTS: Dexlansoprazole MR 60 and 90 mg were superior to placebo for maintaining healing (P < 0.0025). Maintenance rates were 87% and 82% for the 60 and 90 mg doses, respectively, vs. 26% for placebo (life table), and 66% and 65% vs. 14%, respectively (crude rate). Both doses were superior to placebo for the percentage of 24-h heartburn-free days (60 mg, 96%; 90 mg, 94%; placebo, 19%) and nights (98%, 97%, and 50%, respectively). Diarrhoea, flatulence, gastritis (symptoms) and abdominal pain occurred more frequently with dexlansoprazole MR than placebo, but were not dose-related. CONCLUSION: Dexlansoprazole MR effectively maintained healed erosive oesophagitis and symptom relief compared with placebo, and was well tolerated. PMID- 19681810 TI - Systematic review: the short-term and long-term efficacy of adalimumab following discontinuation of infliximab. AB - BACKGROUND: Therapy with adalimumab has been shown to be effective in Crohn's disease (CD) patients who have lost response or are intolerant to infliximab. AIM: To determine the efficacy of adalimumab in CD patients who discontinued infliximab through a systematic review. METHODS: Electronic searches of EMBASE and MEDLINE databases up to May 1, 2009, as well as abstracts from the AGA (2006 2008), ACG (2006-2007), UEGW (2006-2008) and CDDW (2006-2009) identified randomized-controlled trials (RCT) or open-labelled cohorts (OLC) evaluating the short-term and/or long-term efficacy of adalimumab in infliximab failures. The response rates for short-term (clinical response and remission at 4 weeks) and long-term (remission at 6 and 12 months) efficacy were considered. RESULTS: A total of 1810 CD patients were identified among the 15 studies (2 RCT and 13 OLC). The majority of studies evaluated CD patients who either lost response or were intolerable to infliximab, although five OLCs permitted patients refractory to infliximab. Short-term clinical response (n = 9 articles) ranged from 41% to 83%. Long-term clinical remission at 12 months (n = 8 articles) ranged from 19% to 68%. The occurrence of severe adverse events ranged from 0% to 19% and four patients died. CONCLUSIONS: Current RCT and OLC evidence suggest that adalimumab is an efficacious therapy for CD patients who discontinue infliximab. PMID- 19681811 TI - Diverticular disease hospital admissions are increasing, with poor outcomes in the elderly and emergency admissions. AB - BACKGROUND: Diverticular disease has a changing disease pattern with limited epidemiological data. AIM: To describe diverticular disease admission rates and associated outcomes through national population study. METHODS: Data were obtained from the English 'Hospital Episode Statistics' database between 1996 and 2006. Primary outcomes examined were 30-day overall and 1-year mortality, 28-day readmission rates and extended length of stay (LOS) beyond the 75th percentile (median inpatient LOS = 6 days). Multiple logistic regression analysis was used to determine independent predictors of these outcomes. RESULTS: Between the study dates 560 281 admissions with a primary diagnosis of diverticular disease were recorded in England. The national admission rate increased from 0.56 to 1.20 per 1000 population/year. 232 047 (41.4%) were inpatient admissions and, of these, 55 519 (23.9%) were elective and 176 528 (76.1%) emergency. Surgery was undertaken in 37 767 (16.3%). The 30-day mortality was 5.1% (n = 6735) and 1-year mortality was 14.5% (n = 11 567). The 28-day readmission rate was 9.6% (n = 21 160). Increasing age, comorbidity and emergency admission were independent predictors of all primary outcomes. CONCLUSIONS: Diverticular disease admissions increased over the course of the study. Patients of increasing age, admitted as emergency and significant comorbidity should be identified, allowing management modification to optimize outcomes. PMID- 19681812 TI - How do living kidney donors develop end-stage renal disease? AB - The clinical course and risk factors for developing end-stage renal disease (ESRD) after heminephrectomy in living kidney donors have scarcely been investigated. We reviewed medical records and identified eight case donors who developed chronic kidney disease (CKD) stage 5 or ESRD, and subsequently investigated the association between postoperative clinical courses and changes in renal function. To conduct a case-control study, we also selected a control group comprising 24 donors who had maintained stable renal function and were matched for age, sex and follow-up time since donation. Except for one donor who developed ESRD caused by a traffic accident, none of the donors developed progressive renal dysfunction immediately after donation. Their renal functions remained stable for a long period of time, but started to decline after developing new comorbidities, especially risk factors known as progression factors (proteinuria or hypertension) or accelerating factors (cardiovascular [CV] event or infection) of CKD. As compared with the control donors, incidence of postoperative persistent proteinuria, acute CV event, severe infection and hospitalization due to accelerating factors of CKD were significantly higher in the case donors. These results suggest the importance of long-term (more than 10 years) follow-up of donors with special attention on the risk factors of CKD. PMID- 19681813 TI - Pharmacokinetics for once- versus twice-daily tacrolimus formulations in de novo kidney transplantation: a randomized, open-label trial. AB - Tacrolimus, a cornerstone immunosuppressant, is widely available as a twice-daily formulation (Tacrolimus BID). A once-daily prolonged-release formulation (Tacrolimus QD) has been developed that may improve adherence and impart long lasting graft protection. This study compared the pharmacokinetics (PK) of tacrolimus in de novo kidney transplant patients treated with Tacrolimus QD or Tacrolimus BID. A 6-week, open-label, randomized comparative study was conducted in centers in Europe and Australia. Eligible patients received Tacrolimus QD or Tacrolimus BID. PK profiles were obtained following the first tacrolimus dose (day 1), and twice under steady-state conditions. As secondary objectives, efficacy and safety parameters were also evaluated. Sixty-six patients completed all PK profiles (34 Tacrolimus QD, 32 Tacrolimus BID). Mean AUC(0-24) of tacrolimus on day 1 was approximately 30% lower for Tacrolimus QD than Tacrolimus BID (232 and 361 ng.h/mL, respectively), but was comparable by day 4. There was a good correlation and a similar relationship between AUC(0-24) and C(min) for both formulations. Efficacy and safety data were also comparable over the 6-week period. Tacrolimus QD can be administered once daily in the morning on the basis of the same systemic exposure and therapeutic drug monitoring concept as Tacrolimus BID. PMID- 19681814 TI - Immunosuppressant therapy adherence and graft failure among pediatric renal transplant recipients. AB - The study objective was to determine the association between immunosuppressant therapy (IST) adherence and graft failure among pediatric renal transplant recipients (RTRs) using data reported in the United States Renal Data System (USRDS), which contains Medicare prescription claims. RTRs ( 0.05). The total thickness of the cartilage layers was also unaffected by the treatment for 7 days (P > 0.05). In particular, there was no change in the thickness of the perichondrium and reserve cell layer at the measured condylar regions (P > 0.05). However, the thickness of the proliferating cell layer was reduced significantly, whereas the thickness of hypertrophied cartilage layer was increased (P < 0.05). The number of chondroclasts engaged in hypertrophied cartilage resorption was reduced significantly by the alendronate treatment (P < 0.05). The level of MMP-9 expression was reduced at both the transcription and translation levels by the alendronate treatment for 7 and 10 days. These results indicate that alendronate (>3.5 mg/kg/week) inhibits the longitudinal growth of the mandibular condyle by inhibiting chondrocyte proliferation and the resorption of hypertrophied cartilage for ossification. PMID- 19681836 TI - Improvement of hemocompatibility in centrifugal blood pump with hydrodynamic bearings and semi-open impeller: in vitro evaluation. AB - We have developed a noncontact-type centrifugal blood pump with hydrodynamic bearings and a semi-open impeller for mechanical circulatory assist. The impeller is levitated by an original spiral-groove thrust bearing and a herringbone-groove journal bearing, without any additional displacement-sensing module or additional complex control circuits. The pump was improved by optimizing the groove direction of the spiral-groove thrust bearing and the pull-up magnetic force between the rotor magnet and the stator coil against the impeller. To evaluate hemocompatibility, we conducted a levitation performance test and in vitro hemocompatibility tests by means of a mock-up circulation loop. In the hemolysis test, the normalized index of hemolysis was reduced from 0.721 to 0.0335 g/100 L corresponding to an expansion of the bearing gap from 1.1 to 56.1 microm. In the in vitro antithrombogenic test, blood pumps with a wide thrust bearing gap were effective in preventing thrombus formation. Through in vitro evaluation tests, we confirmed that hemocompatibility was improved by balancing the hydrodynamic fluid dynamics and magnetic forces. PMID- 19681837 TI - Dispersion and accuracy of simulated phosphene positioning using tactile board. AB - The evaluation of phosphene is one of the most important things after the electrode array's implantation. Some experiments have been used to study the basic rules on people using their tactile perception in describing the characteristics of simulated phosphenes, such as the dispersion, accuracy, and response time. In order to describe the simulated phosphenes more systematically and scientifically, an improved phosphene-positioning method using tactile board was designed in this study. Using tactile board to guide the tactile perception, the dispersion was limited to less than 6 mm, while the largest mean error was 8.1 mm, which was nearly equal to the minimal resolution of tactile board (8 mm). The response time greatly increased because of the standard processes in using tactile board. Moreover, the long-term experiment to repeat the procedure could improve and sustain the subjects' performance in good results. PMID- 19681838 TI - Extracorporeal life support as ultimate strategy for refractory severe cardiogenic shock induced by Tako-tsubo cardiomyopathy: a new effective therapeutic option. AB - We report a possible new therapeutic strategy, using extracorporeal cardiopulmonary support (ECLS), for severe refractory cardiogenic shock (SRCS) in a patient with Tako-tsubo cardiomyopathy (TC). TC is a syndrome characterized by left ventricular wall motion abnormalities, without coronary artery disease, mimicking the diagnosis of acute coronary syndrome. This ventricular dysfunction can be reversible; however, it can progress into refractory cardiogenic shock with limited therapeutic options available. For the first time in a Tako-tsubo patient with refractory cardiogenic shock, we used ECLS treatment in order to rest the heart, sustain circulation and end-organ perfusion, and promote potential ventricular recovery. ECLS might be the selected treatment for SRCS in patients with TC, and seems to be an effective and useful ultimate therapeutic strategy for preventing death. PMID- 19681839 TI - Coronary arteries angiogenesis in ischemic myocardium: biocompatibility and biodegradability of various hydrogels. AB - To evaluate the biocompatibility and biodegradability of various hydrogels and choose suitable hydrogels for the coronary arteries angiogenesis in ischemic myocardium, we synthesized six kinds of hyaluronan hydrogels, fibrin hydrogel, poly(vinyl alcohol)-chitosan hydrogel, and obtained elastin hydrogels. We examined their degradation rates and cytotoxicity in vitro. Then, hydrogels were implanted into rat adductor muscles for 1, 2, or 4 weeks. Hydrogels and surrounding tissues were resected, followed by hematoxylin and eosin staining, Masson's trichrome staining, and immunohistochemical staining for measurements of degradation, immune response, and angiogenesis. 2-Iminothiolane grafted hyaluronan hydrogel and periodate oxidated hyaluronan hydrogel presented rapid degradation rates, low quantity of inflammation-mediating cells (12 +/- 3 and 12 +/- 4 per 2.5 x 10(-3) mm(2), respectively, at week 2), thin fibrous capsules (scores were 3.8 +/- 0.1 and 4.0 +/- 0.3 per 0.33 mm(2), respectively, at week 2) with dense blood vessels in the areas surrounding the implanted hydrogels. 2 Iminothiolane grafted hyaluronan and periodate oxidated hyaluronan hydrogels with appropriate degradation rates and low immune responses were suitable for coronary arteries angiogenesis in ischemic myocardium. PMID- 19681840 TI - Association among serum fetuin-A level, coronary artery calcification, and bone mineral densitometry in maintenance hemodialysis patients. AB - Patients with end-stage renal disease have a very high prevalence and extent of arterial calcification. A number of studies suggest that similar pathophysiologic mechanisms are responsible for development and progression of calcification of atherosclerotic plaque and bone formation. Fetuin-A is a potent calcification inhibitor and is expressed in bone, with not-yet well-defined functions. The aim of this study was to investigate the relation between bone mineral densitometry parameters, coronary artery calcification, and serum fetuin-A levels. In a cross sectional design, we included 72 maintenance hemodialysis (HD) patients and 30 age- and gender-matched healthy controls. Serum fetuin-A levels were studied both in maintenance HD patients and healthy controls. Maintenance HD patients had radius, hip, and lumbar spine bone mineral density (BMD) assessed by dual-energy X-ray absorptiometry and coronary artery calcification score (CACS) measured by electron-beam computed tomography. The associations between site-specific BMD parameters, CACS, and serum fetuin-A levels were studied in maintenance HD patients. CACS, mass, and volume of plaques in coronary arteries were significantly higher in patients with a T-score below -2.5 than above in the proximal region of the radius, neck and trochanter of the femur, and the lumbar spine. Mean serum fetuin-A concentration was 0.636 +/- 0.118 g/L in maintenance HD patients and it was less than healthy controls (0.829 +/- 0.100 g/L, P < 0.0001). CACS, mass, and volume of plaques in coronary arteries correlated significantly with the serum fetuin-A levels. Moreover, significant positive correlations were shown between the serum fetuin-A levels, BMD values, and T scores of proximal radius, neck, and trochanter of the femur, but not with the lumbar spine. The present study demonstrates an association between serum fetuin A levels, coronary artery calcification, and bone mineral densities--except for the lumbar spine, in maintenance HD patients. However, the results should be interpreted with caution because of the cross-sectional design of the study. PMID- 19681841 TI - Myoelectric hand prosthesis force control through servo motor current feedback. AB - This paper presents the prehension force closed-loop control design of a mechanical finger commanded by electromyographic signal (EMG) from a patient's arm. The control scheme was implemented and tested in a mechanical finger prototype with three degrees of freedom and one actuator, driven by arm muscles EMG of normal volunteers. Real-time indirect estimation of prehension force was assessed by measuring the DC servo motor actuator current. A model of the plant comprising finger, motor, and grasped object was proposed. Model parameters were identified experimentally and a classical feedback phase-lead compensator was designed. The controlled mechanical finger was able to provide a more accurate prehension force modulation of a compliant object when compared to open-loop control. PMID- 19681842 TI - Computational design and in vitro characterization of an integrated maglev pump oxygenator. AB - For the need for respiratory support for patients with acute or chronic lung diseases to be addressed, a novel integrated maglev pump-oxygenator (IMPO) is being developed as a respiratory assist device. IMPO was conceptualized to combine a magnetically levitated pump/rotor with uniquely configured hollow fiber membranes to create an assembly-free, ultracompact system. IMPO is a self contained blood pump and oxygenator assembly to enable rapid deployment for patients requiring respiratory support or circulatory support. In this study, computational fluid dynamics (CFD) and computer-aided design were conducted to design and optimize the hemodynamics, gas transfer, and hemocompatibility performances of this novel device. In parallel, in vitro experiments including hydrodynamic, gas transfer, and hemolysis measurements were conducted to evaluate the performance of IMPO. Computational results from CFD analysis were compared with experimental data collected from in vitro evaluation of the IMPO. The CFD simulation demonstrated a well-behaved and streamlined flow field in the main components of this device. The results of hydrodynamic performance, oxygen transfer, and hemolysis predicted by computational simulation, along with the in vitro experimental data, indicate that this pump-lung device can provide the total respiratory need of an adult with lung failure, with a low hemolysis rate at the targeted operating condition. These detailed CFD designs and analyses can provide valuable guidance for further optimization of this IMPO for long-term use. PMID- 19681843 TI - Model-based analysis of potassium removal during hemodialysis. AB - Potassium ion (K(+)) kinetics in intra- and extracellular compartments during dialysis was studied by means of a double-pool computer model, which included potassium-dependent active transport (Na-K-ATPase pump) in 38 patients undergoing chronic hemodialysis. Each patient was treated for 2 weeks with a constant K(+) dialysate concentration (K(+)(CONST) therapy) and afterward for 2 weeks with a time-varying (profiled) K(+) dialysate concentration (K(+)(PROF) therapy). The two therapies induced different levels of K(+) plasma concentration (K(+)(CONST): 3.71 +/- 0.88 mmol/L vs. K(+)(PROF): 3.97 +/- 0.64 mmol/L, time-averaged values, P < 0.01). The computer model was tuned to accurately fit plasmatic K(+) measured in the course and 1 h after K(+)(CONST) and K(+)(PROF) therapies and was then used to simulate the kinetics of intra- and extracellular K(+). Model-based analysis showed that almost all the K(+) removal in the first 90 min of dialysis was derived from the extracellular compartment. The different K(+) time course in the dialysate and the consequently different Na-K pump activity resulted in a different sharing of removed potassium mass at the end of dialysis: 56% +/- 17% from the extracellular compartment in K(+)(PROF) versus 41% +/- 14% in K(+)(CONST). At the end of both therapies, the K(+) distribution was largely unbalanced, and, in the next 3 h, K(+) continued to flow in the extracellular space (about 24 mmol). After rebalancing, about 80% of the K(+) mass that was removed derived from the intracellular compartment. In conclusion, the Na-K pump plays a major role in K(+) apportionment between extracellular and intracellular compartments, and potassium dialysate concentration strongly influences pump activity. PMID- 19681844 TI - The role of interspecific interference competition in character displacement and the evolution of competitor recognition. AB - The extent to which interspecific interference competition has contributed to character evolution is one of the most neglected problems in evolutionary biology. When formerly allopatric species come into secondary contact, aggressive interactions between the species can cause selection on traits that affect interspecific encounter rates (e.g. habitat preferences, activity schedules), competitor recognition (e.g. colouration, song), and fighting ability (e.g. weaponry, body size). We define agonistic character displacement (ACD) as the process of phenotypic evolution in a population caused by interference competition with one or more sympatric species and which results in shifts in traits that affect the rate, intensity or outcome of interspecific aggression. After clarifying the relationships between ACD and other evolutionary processes that may occur when species come into secondary contact, we develop an individual based, quantitative genetic model to examine how traits involved in competitor recognition would be expected to evolve under different secondary contact scenarios. Our simulation results show that both divergence and convergence are possible outcomes, depending on the intensity of interspecific exploitative competition, the costs associated with mutual versus unilateral recognition, and the extent of phenotypic differences prior to secondary contact. We then devise a set of eight criteria for evaluating putative examples of ACD and review the empirical literature to assess the strength of existing evidence and to identify promising avenues for future research. Our literature search revealed 33 putative examples of ACD across insects, fishes, bats, birds, lizards, and amphibians (15 divergence examples; 18 convergence examples). Only one example satisfies all eight criteria for demonstrating ACD, but most case studies satisfy four or more criteria. The current state of the evidence for ACD is similar to the state of the evidence for ecological character displacement just 10 years ago. We conclude by offering suggestions for further theoretical and empirical research on ACD. PMID- 19681845 TI - The recurrence risk of severe de novo pre-eclampsia in singleton pregnancies: a population-based cohort. AB - OBJECTIVE: Previous studies have found recurrence risks of severe pre-eclampsia as high as 40%. Our objective was to determine both the recurrence risk of severe de novo pre-eclampsia and risk factors associated with it in a contemporaneous population. STUDY DESIGN: Population-based retrospective cohort study. POPULATION: Women who had two or more singleton liveborn or stillborn hospital deliveries in Ontario, Canada between April 1994 and March 2002 and without a history of chronic hypertension. METHODS: International Classification of Disease codes were used to identify patients in the Canadian Institute for Health Information Discharge Abstract Database. MAIN OUTCOME MEASURES: The absolute and adjusted risks of recurrent severe de novo pre-eclampsia were determined. RESULTS: Between 1 April 1994 and 30 March 2002, there were 185 098 women with two or more singleton deliveries >20 weeks in the province of Ontario, Canada. There were 1954 women who had severe de novo pre-eclampsia in the index pregnancy, 133 of whom had recurrent severe pre-eclampsia, for a risk of recurrent severe pre-eclampsia of 6.8% (95% CI 5.7-7.9%). The risk of recurrent severe de novo pre-eclampsia was increased in women with pre-existing renal disease (adjusted OR 17.98, 95% CI 3.50-92.52) and those >35 years of age (adjusted OR 3.79, 95% CI 2.04-7.04, reference 20-25 years). CONCLUSIONS: The recurrence risk of severe de novo pre-eclampsia in our population-based cohort study (6.8%) is lower than previously published reports in selected populations. PMID- 19681846 TI - Association of interferon-gamma +874A polymorphism with the risk of developing cervical cancer in north-Indian population. AB - OBJECTIVE: Interferon gamma (IFN-gamma) is a pro-inflammatory cytokine playing a pivotal role in both innate and adaptive immune responses. A single nucleotide polymorphism located in the first intron of the human IFN-gamma gene can influence the secretion of cytokine. Therefore, we aimed to investigate the association of IFN-gamma T/A gene polymorphism with the risk of cervical cancer. DESIGN: Case-control study. SETTING: Uttar Pradesh State in India. SAMPLE: Two hundred cases with histologically proven cancer of the cervix and healthy controls (n = 230), age and ethnicity matched were recruited in this study. METHODS: Genotyping was performed for bi-allelic +874 (T/A) polymorphism of IFN gamma by amplification refractory mutation system method. MAIN OUTCOME MEASURES: Low producer IFN-gamma +874 AA genotype was associated with high risk for cervical cancer, which further modulated the increased risk in tobacco users. RESULTS: IFN-gamma AA genotype which is low producer of IFN-gamma was associated with increased risk of cervical cancer (OR = 2.43, P = 0.003). Allele A was at 1.54-fold increased risk of cervical cancer (OR=1.54, P = 0.002). The AA genotype showed statistically significant risk with high stage (III + IV) of cervical cancer (OR = 4.99, P = 0.001). In tobacco users, AA genotype showed significantly increased susceptibility to cervical cancer (OR = 5.08, P = 0.010). CONCLUSION: Variation in IFN-gamma +874 AA genotype because of ethnicity in north-Indian population may represent an important susceptibility biomarker for cervical cancer risk as well as other diseases and should be explored further. PMID- 19681847 TI - Urethrovaginal fistula associated with 'sterile abscess' formation following periurethral injection of dextranomer/hyaluronic acid co-polymer (Zuidex) for the treatment of stress urinary incontinence--a case report. PMID- 19681848 TI - Visceral hyperalgesia in chronic pelvic pain. PMID- 19681849 TI - Comparison of pregnancy outcomes in women with repaired versus unrepaired atrial septal defect. AB - OBJECTIVE: To compare the risks of complications during pregnancy in women with repaired and unrepaired atrial septal defects (ASDs) without associated complex cardiac lesions. DESIGN: A retrospective multicentre study. SETTING: Tertiary centres in the Netherlands and Belgium. POPULATION: Women with ASD without associated complex cardiac lesions. METHODS: Women were identified using two congenital heart disease registries. One hundred women were identified who had 243 pregnancies, including 49 miscarriages and six terminations of pregnancy. Detailed information on each completed pregnancy (n = 188; unrepaired ASD, n = 133; repaired ASD, n = 55) was obtained using medical records and telephone interviews. In addition, data from the Generation R database (a prospective cohort study; n = 9667) were used to determine the background risk (control group). MAIN OUTCOME MEASURES: Adjusted odds ratios (AORs) for cardiac, obstetric and neonatal events controlled for multiple pregnancies per woman using general estimating equation analysis. RESULTS: Women with an unrepaired ASD had a higher risk of neonatal events (AOR = 2.99, 95% confidence interval [CI] 1.14-7.89, P = 0.027) than women with a repaired ASD. The risk of cardiac and obstetric complications was comparable between women with unrepaired and repaired ASDs. Compared with the general population, women with an unrepaired ASD had higher risks of pre-eclampsia (AOR = 3.54, 95% CI 1.26-9.98, P = 0.017), small-for gestational-age births (AOR = 1.95, 95% CI 1.15-3.30, P = 0.013) and fetal mortality (AOR = 5.55, 95% CI 1.77-17.4, P = 0.003). By contrast, no differences were observed when comparing women with a repaired ASD versus controls. CONCLUSIONS: Women with an unrepaired ASD are at increased risk of neonatal events in comparison with women with a repaired ASD. Compared with the general population, women with an unrepaired ASD are at increased risk of pre-eclampsia, small-for-gestational-age births and fetal mortality. PMID- 19681850 TI - Maternal complications and pregnancy outcome in women with mechanical prosthetic heart valves treated with enoxaparin. AB - OBJECTIVE: To determine maternal and fetal outcomes in women with mechanical heart valves managed with therapeutic dose enoxaparin during pregnancy. DESIGN: Retrospective audit. SETTING: Hospital-based high-risk antenatal clinics. POPULATION: Pregnant women with mechanical heart valves attending high-risk antenatal clinics, treated with enoxaparin (1 mg/kg twice daily) during pregnancy. METHODS: Women with mechanical heart valves treated with enoxaparin at any stage during pregnancy (1997-2008) identified using a database of women with mechanical heart valves attending the high-risk clinics and a prospective database of women prescribed enoxaparin for any indication during pregnancy. MAIN OUTCOME MEASURES: Maternal outcomes included thromboembolic and haemorrhagic complications. Pregnancy and fetal outcomes included miscarriage, stillbirth, baby death and live birth, small-for-gestational-age infants, warfarin embryopathy and warfarin-related fetal loss. RESULTS: Thirty-one women underwent 47 pregnancies. In 34 pregnancies (72.3%), anticoagulation was with predominantly enoxaparin and 13 (27.7%) pregnancies women received mainly warfarin, with enoxaparin given in the first trimester and/or peri-delivery. Seven (14.9%) thrombotic complications occurred, of which five (10.6%) were associated with enoxaparin treatment. Non-compliance or sub-therapeutic anti-Xa levels contributed in each case. Antenatal and postpartum haemorrhagic complications occurred in eight (17%) and 15 (32%) pregnancies respectively. Of 35 pregnancies continuing after 20 weeks' gestation, 96% (22/23) of women taking predominantly enoxaparin had a surviving infant compared with 75% (9/12) in women taking primarily warfarin. Four perinatal deaths occurred, three attributable to warfarin. CONCLUSIONS: Compliance with therapeutic dose enoxaparin and aspirin during pregnancy in women with mechanical heart valves is associated with a low risk of valve thrombosis and good fetal outcomes, but close monitoring is essential. PMID- 19681851 TI - Haemostatic changes in the puerperium '6 weeks postpartum' (HIP Study) - implication for maternal thromboembolism. AB - OBJECTIVE: We aim to measure the thrombotic changes during the postnatal period up to 6 weeks after delivery and assess the extent of the risk period. DESIGN: Prospective observational study. SETTING: Queen Elizabeth II, an acute District General Hospital, Hertfordshire. POPULATION: Women booked at the antenatal clinic and prepared to deliver at the hospital. METHODS: We assessed the haemoglobin, platelet count and function, fibrinogen, prothrombin time, activated partial thromboplastin time, protein C, S and antithrombin level and as well as rotational thromboelastometry (ROTEM) from predelivery till 6 weeks postpartum. RESULTS: A total 50 women were recruited of which four dropped out. Results compared against the finding at 6 weeks after delivery. Platelet was significantly elevated on day 19 compared to day 42 (P < 0.001). Fibrinogen was elevated from predelivery till day 15 after delivery (P < 0.01). Prothrombin time (PT) was low till day 15 (P < 0.05) and activated partial thromboplastin time (APTT) was significantly lower till day 3 after delivery (P < 0.001). ROTEM revealed low clotting time (CT) at predelivery and continued to be low till day 7. Clot formation time (CFT) significantly low till day 25 (P < 0.05). Maximum clot firmness, alpha angle and amplitude at 20 minutes were raised till day 19 (P < 0.001, P < 0.01 and P < 0.001 respectively). While, comparing vaginal delivery against caesarean section there were nonsignificant increase in thrombotic parameters in caesarean section. CONCLUSION: Coagulation screens as well as thomboelastometry suggest a persistent hypercoagulation during the first 3 weeks after delivery. PMID- 19681853 TI - Vitamin D in pregnancy-time for action: a paediatric audit. AB - Maternal vitamin D deficiency is a significant public health issue. This retrospective multicentre audit was undertaken to identify the scale of maternal vitamin D deficiency in London, using infant vitamin D deficiency as a surrogate marker. During January 2006 to June 2008, 74 infants presented with symptomatic vitamin D deficiency, a prevalence of 1.6 per 1000 deliveries in London. Of these, 49% were hypocalcaemic at presentation and 27% had hypocalcaemic seizures. A telephone survey in June 2008 showed that no London National Health Service antenatal units had departmental guidelines on vitamin D. This audit is presented to raise awareness of current recommendations and the sequelae of maternal vitamin D deficiency. PMID- 19681852 TI - Association between antidepressant drug use during pregnancy and child healthcare utilisation. AB - OBJECTIVE: To evaluate healthcare utilisation by children who were exposed to antidepressant drug use during pregnancy and those whose mothers stopped using antidepressants before pregnancy compared with a control group. DESIGN: Cohort study. Setting Health insurance records in the Netherlands. POPULATION: A total of 38 602 children born between 2000 and 2005. METHODS: Survey of child healthcare utilisation in relation to gestational antidepressant use. MAIN OUTCOME MEASURE: Healthcare utilisation rates during the first year of life, with special emphasis to medical care related to cardiac disease. RESULTS: Children of mothers who used antidepressants during pregnancy showed increased healthcare use during the first year of life, independent of the mother's healthcare use. The relative risk of more than two visits to general practitioners was 1.5 (95% confidence interval, CI: 1.3-1.8) in the continuous antidepressant users group and 1.3 (95% CI: 1.2-1.5) in the group of children whose mothers stopped taking medication. In both study groups there was a trend towards more drug use for infections and inflammation compared with the control group. Children continuously exposed to antidepressants had an increased risk of cardiac interventions such as cardiovascular surgery or heart catheterisation, relative risk of 5.6 (95% CI: 1.8-17.4). The risk of physiotherapy was twice as high in the antidepressant group compared with the control group (relative risk 2.0; 95% CI: 1.5-2.6). CONCLUSION: Antidepressant use during pregnancy is associated with increased child healthcare utilisation and increased risk of major cardiac interventions in early childhood. PMID- 19681854 TI - Noninvasive methods of detecting fetal anaemia: a systematic review and meta analysis. AB - OBJECTIVES: To estimate in a systematic review of the literature the diagnostic value of ultrasound and Doppler blood flow velocity in the evaluation of fetal anaemia. STUDY SELECTION AND DATA EXTRACTION: Literature from 2000 to 2008 was identified using MEDLINE and EMBASE, the Cochrane Library and relevant specialist register of the Cochrane Collaboration, and by checking reference lists of known primary studies and review articles. Studies were selected if the accuracy of the fetal ultrasound parameters or Doppler studies of blood flow in the fetal vessels was estimated compared with a reference standard. Data from the selected studies were abstracted as 2 x 2 tables comparing the diagnostic test result with the reference standard. Results were pooled where appropriate. Diagnostic accuracy was expressed as likelihood ratios. RESULTS: Twenty-five primary studies were identified containing suitable data on middle cerebral artery Doppler peak systolic velocity (MCA-PSV). The largest group of studies whose data could be pooled containing nine studies gave a positive likelihood ratio of 4.30 (95% CI: 2.50 to 7.41) and a negative likelihood ratio of 0.30 (95% CI: 0.13 to 0.69) for 675 cases in detecting severe anaemia in the analysis. DISCUSSION: Although middle cerebral artery peak systolic velocity Doppler has limited diagnostic accuracy, it remains the gold standard for noninvasive screening of fetal anaemia. PMID- 19681855 TI - An eclipse in the subungual space: a diagnostic sign for a subungual epidermal cyst? PMID- 19681856 TI - Absolute CD3+ CD158k+ lymphocyte count is reliable and more sensitive than cytomorphology to evaluate blood tumour burden in Sezary syndrome. AB - Background CD158k/KIR3DL2 is a specific marker for Sezary cells which can be used to diagnose Sezary syndrome (SS) in erythrodermic patients with abnormal circulating T cells. Objectives To evaluate the suitability of CD158k/KIR3DL2 for detecting and evaluating blood tumour load during the follow up of patients with SS. Methods The absolute CD3+ CD158k+ lymphocyte count was compared with the absolute count of cytomorphological Sezary cells and was correlated with clinical flares in a cohort of patients with SS. Twenty-five patients were included in the study and 48 blood samples were analysed. Results The absolute count of CD3+ CD158k+ cells strongly correlated with the absolute count of atypical circulating cells (r = 0.97, P < 10(-15)). The CD3+ CD158k+ lymphocyte cell count was in eight cases more sensitive than cytomorphology for detecting atypical circulating cells especially for small-sized tumour cells. The tumour burden evaluated by CD3+ CD158k+ immunostaining was significantly associated with clinical flare (P < 10(-4)). Conclusions CD3+ CD158k+ phenotyping is a reliable and objective test to monitor the blood tumour burden in patients with SS under systemic therapy. PMID- 19681857 TI - A population-based study of acute medical conditions associated with bullous pemphigoid. AB - BACKGROUND: Bullous pemphigoid is associated with poorly understood dramatically increased early mortality rates. OBJECTIVES: To assess the incidence of acute events predisposing to early mortality. METHODS: Computerized medical records from the Health Improvement Network, a large population-based U.K. general practice database, were used to conduct a cohort analysis. Outcome measures were incidence rates of myocardial infarction, pulmonary embolism, pneumonia and sepsis compared with a matched control population. RESULTS: People with bullous pemphigoid were three times as likely to develop pneumonia, adjusted rate ratio 2.94 [95% confidence interval (CI) 2.01-4.31] or pulmonary embolism, adjusted rate ratio 3.12 (95% CI 1.37-7.12) compared with matched controls. No statistically significant increase was seen for myocardial infarction, adjusted rate ratio 1.24 (95% CI 0.66-2.33), or sepsis, adjusted rate ratio 2.02 (95% CI 0.78-5.21). CONCLUSIONS: The risk of pulmonary embolism and pneumonia is increased following a diagnosis of bullous pemphigoid. It may be possible to reduce associated mortality through considering prophylaxis with either antithromboembolic measures or antibiotic therapy and vaccination. PMID- 19681858 TI - Generalized pustular psoriasis induced by systemic glucocorticosteroids: four cases and recommendations for treatment. PMID- 19681860 TI - Filaggrin haploinsufficiency is highly penetrant and is associated with increased severity of eczema: further delineation of the skin phenotype in a prospective epidemiological study of 792 school children. AB - BACKGROUND: Null mutations within the filaggrin gene (FLG) cause ichthyosis vulgaris and are associated with atopic eczema. However, the dermatological features of filaggrin haploinsufficiency have not been clearly defined. OBJECTIVES: This study investigated the genotype-phenotype association between detailed skin phenotype and FLG genotype data in a population-based cohort of children. METHODS: Children (n = 792) aged 7-9 years were examined by a dermatologist. Features of ichthyosis vulgaris, atopic eczema and xerosis were recorded and eczema severity graded using the Three Item Severity score. Each child was genotyped for the six most prevalent FLG null mutations (R501X, 2282del4, R2447X, S3247X, 3702delG, 3673delC). Fisher's exact test was used to compare genotype frequencies in phenotype groups; logistic regression analysis was used to estimate odds ratios and penetrance of the FLG null genotype and a permutation test performed to investigate eczema severity in different genotype groups. RESULTS: Ten children in this cohort had ichthyosis vulgaris, of whom five had mild-moderate eczema. The penetrance of FLG null mutations with respect to flexural eczema was 55.6% in individuals with two mutations, 16.3% in individuals with one mutation and 14.2% in wild-type individuals. Summating skin features known to be associated with FLG null mutations (ichthyosis, keratosis pilaris, palmar hyperlinearity and flexural eczema) showed a penetrance of 100% in children with two FLG mutations, 87.8% in children with one FLG mutation and 46.5% in wild-type individuals (P < 0.0001, Fisher exact test). FLG null mutations were associated with more severe eczema (P = 0.0042) but the mean difference was only 1-2 points in severity score. Three distinct patterns of palmar hyperlinearity were observed and these are reported for the first time. CONCLUSIONS: Filaggrin haploinsufficiency appears to be highly penetrant when all relevant skin features are included in the analysis. FLG null mutations are associated with more severe eczema, but the effect size is small in a population setting. PMID- 19681861 TI - Forty-two novel COL7A1 mutations and the role of a frequent single nucleotide polymorphism in the MMP1 promoter in modulation of disease severity in a large European dystrophic epidermolysis bullosa cohort. AB - BACKGROUND: Dystrophic epidermolysis bullosa (DEB) is a severe genetic skin blistering disorder caused by mutations in the gene COL7A1, encoding collagen VII. Recently, the MMP1 promoter single nucleotide polymorphism (SNP) rs1799750, designated as 1G 2G, was shown to be involved in modulation of disease severity in patients with recessive DEB (RDEB), and was proposed as a genetic modifier. OBJECTIVES: To identify the molecular basis of DEB in 103 individuals and to replicate the results of the MMP1 promoter SNP analysis in an independent patient group, as verification is necessary in such a rare and heterogeneous disorder. METHODS: To determine the molecular basis of the disease, we performed COL7A1 mutation screening, reverse transcription-polymerase chain reaction (PCR) and real-time quantitative PCR. The status of the MMP1 SNP was analysed by PCR and restriction enzyme digestion and verified by sequencing. RESULTS: We disclosed 42 novel COL7A1 mutations, including the first large genomic deletion of 4 kb affecting only the COL7A1 gene, and three apparently silent mutations affecting splicing. Even though the frequency of the high-risk allele was increased in patients with RDEB, no statistically significant correlation between disease severity and genotype could be made. Also, no correlation was observed with development of squamous cell carcinoma, a severe complication of DEB. CONCLUSIONS: Taken together, the results suggest that the MMP1 SNP is not the sole disease modifier in different forms of DEB, and other genetic and environmental factors contribute to the clinical phenotype. PMID- 19681859 TI - Tolerability of high-dose topical tretinoin: the Veterans Affairs Topical Tretinoin Chemoprevention Trial. AB - BACKGROUND: Topical tretinoin is a medication commonly used for acne that has potential application in the long-term treatment of photodamaged skin. However, there are few published data regarding the tolerability of high-dose tretinoin with long-term use. OBJECTIVES: To assess the long-term tolerability of tretinoin 0.1% cream. METHODS: A randomized, multicentre, double-blind, controlled trial for chemoprevention of keratinocyte carcinomas (i.e. basal cell or squamous cell carcinomas) using topical tretinoin cream to the face and ears was conducted. All participants were veterans and had a history of two or more keratinocyte carcinomas over the previous 5 years. Participants were examined (by a study dermatologist) and interviewed every 6 months (for up to 5.5 years to May 2004). Treatment comprised tretinoin 0.1% cream or vehicle control cream once daily, then twice daily as tolerated. Participants were instructed to step down application frequency to once daily or less if twice daily was not tolerated. The main outcome measures were reported side-effects, frequency of cream application and attendance at study visits. Appropriate data were available for four of the six clinical sites of this trial. RESULTS: Data from 736 randomized participants (mean age 71 years; 97% men) from four clinical sites were analysed. The tretinoin group more commonly reported one or more side-effects at the 6-month follow-up than the control group (61% vs. 42%, P < 0.0001). Side-effects decreased over time in both groups, but to a greater extent in the tretinoin group, and the difference became nonsignificant at 30 months. Burning was the most common side-effect (39% tretinoin vs. 17% control, P < 0.0001). There was no difference in severity of side-effects among those affected. Of the participants who reported burning in either group, most reported mild burning; only 11% of those with burning in the tretinoin group reported it as severe (mild 62% tretinoin vs. 70% placebo; severe 11% vs. 5%; P = 0.4). Itching (24% vs. 16%, P = 0.01) and other local cutaneous reactions (12% vs. 6%, P = 0.01) were also more commonly reported by the tretinoin group at 6 months. There was no difference in numbness (2% vs. 2%, P = 0.9). Participants in the tretinoin group were less likely to apply cream twice daily at 6 months (29% vs. 43%, P = 0.0002). This difference persisted over the entire duration of follow-up. There was little difference between groups in attendance at study visits or completion of telephone interviews (92% vs. 95%, P = 0.06). No unexpected adverse events were reported. CONCLUSIONS: Overall, the tolerability level of topical tretinoin was high in this study population, with almost 40% of the tretinoin group reporting no side-effects, and the majority (67%) tolerating at least once-daily dosing at 6-month follow-up. High-dose topical tretinoin is feasible for long-term use in this population. PMID- 19681862 TI - A randomized controlled trial of R-salbutamol for topical treatment of discoid lupus erythematosus. AB - BACKGROUND: In a recent open pilot trial, R-salbutamol sulphate, a well-known molecule with anti-inflammatory effects, was tested successfully on patients with therapy-resistant discoid lupus erythematosus (DLE). OBJECTIVES: To compare the efficacy and safety of R-salbutamol cream 0.5% vs. placebo on DLE lesions in a multicentre, double-blinded, randomized, placebo-controlled phase II trial. METHODS: Thirty-seven patients with at least one newly developed DLE lesion were randomized - 19 to the R-salbutamol cream 0.5% and 18 to placebo - and treated twice daily for 8 weeks. Efficacy was evaluated through scores of erythema, scaling/hypertrophy and induration as well as pain and itching; general improvement scored by the investigator and global improvement scored by patients' assessment were also evaluated. RESULTS: The mean area under the curve of improvement for scaling/hypertrophy, pain, itching and global patient assessment was significantly better for the actively treated patients as compared with placebo (scaling/hypertrophy, P = 0.0262; pain, P = 0.0238; itching, P = 0.0135; global patient assessment, P = 0.045). Moreover, the percentage of patients without induration was significantly higher in the active group compared with the placebo group (P = 0.013), and a statistically significantly greater decrease in the size of the lesional area was also seen in the overall analysis of the R salbutamol-treated patients (P = 0.0197). No serious adverse events were reported. CONCLUSIONS: Application of R-salbutamol cream 0.5% was safe and well tolerated. Statistically significant effects were seen on scaling/hypertrophy, induration, pain and itching as well as patient global assessment, suggesting that R-salbutamol could be a promising new topical therapy alternative for DLE. PMID- 19681863 TI - Cytokine imbalance with increased production of interferon-alpha in psoriasiform eruptions associated with antitumour necrosis factor-alpha treatments. AB - BACKGROUND: Psoriasiform eruptions occur in association with antitumour necrosis factor (TNF)-alpha treatments in autoinflammatory diseases. The major reported clinical presentation is palmoplantar pustulosis, sometimes accompanied with plaque-like psoriasis. In some reports, histological findings suggest psoriasis whereas others favour a lichenoid drug reaction. We present a case series with a comprehensive clinical, histopathological and immunohistochemical study. OBJECTIVES: To investigate the mechanism involved in psoriasiform eruptions in patients receiving anti-TNF-alpha inhibitors. METHODS: Between July 2004 and May 2008, 13 patients with psoriasiform eruptions arising under anti-TNF-alpha treatment were enrolled in the study. Punch biopsy specimens of lesions were processed for standard and immunohistochemical analyses using antibodies against CD3, CD4, CD8, CD20, CD1a, KP1, CXCR3, CXCL9, Tia1 and MxA, which is specifically induced by type I interferons (IFNs). Additionally, we analysed biopsies from lesional skin of patients with cutaneous discoid lupus erythematosus, lichen planus and psoriasis. Control biopsies were taken from unaffected skin. RESULTS: All patients developed psoriasiform plaques on the body accompanied with palmoplantar keratoderma or pustulosis in three patients. Histological and immunohistochemical findings showed a psoriasiform pattern with focal lichenoid and spongiotic features. We detected strong production of the MxA protein in inflammatory cells, indicating involvement of type I IFNs, and the expression was higher than in control psoriasis samples. Expression of MxA was closely associated with the recruitment of CXCR3+ lymphocytes in the skin bearing markers of cytotoxic capacity. CONCLUSIONS: Results support the hypothesis that psoriasiform eruptions are a new model of drug reaction characterized by an increased expression of type I IFNs induced by anti-TNF-alpha. PMID- 19681864 TI - Dual functionality of interleukin-1 family cytokines: implications for anti interleukin-1 therapy. AB - Dysregulated inflammation contributes to disease pathogenesis in both the periphery and the brain. Cytokines are coordinators of inflammation and were originally defined as secreted mediators, released from expressing cells to activate plasma membrane receptors on responsive cells. However, a group of cytokines is now recognized as having dual functionality. In addition to their extracellular effects, these cytokines act inside the nuclei of cytokine expressing or cytokine-responsive cells. Interleukin-1 (IL-1) family cytokines are key pro-inflammatory mediators, and blockade of the IL-1 system in inflammatory diseases is an attractive therapeutic goal. All current therapies target IL-1 extracellular actions. Here we review evidence that suggests IL-1 family members have dual functionality. Several IL-1 family members have been detected inside the nuclei of IL-1-expressing or IL-1-responsive cells, and intranuclear IL-1 is reported to regulate gene transcription and mRNA splicing. However, further work is required to determine the impact of IL-1 intranuclear actions on disease pathogenesis. The intranuclear actions of IL-1 family members represent a new and potentially important area of IL-1 biology and may have implications for the future development of anti-IL-1 therapies. PMID- 19681865 TI - The K+ channels K(Ca)3.1 and K(v)1.3 as novel targets for asthma therapy. AB - Asthma affects 10% of the UK population and is an important cause of morbidity and mortality at all ages. Current treatments are either ineffective or carry unacceptable side effects for a number of patients; in consequence, development of new approaches to therapy are important. Ion channels are emerging as attractive therapeutic targets in a variety of non-excitable cells. Ion channels conducting K(+) modulate the activity of several structural and inflammatory cells which play important roles in the pathophysiology of asthma. Two channels of particular interest are the voltage-gated K(+) channel K(v)1.3 and the intermediate conductance Ca(2+)-activated K(+) channel K(Ca)3.1 (also known as IK(Ca)1 or SK4). K(v)1.3 is expressed in IFNgamma-producing T cells while K(Ca)3.1 is expressed in T cells, mast cells, macrophages, airway smooth muscle cells, fibroblasts and epithelial cells. Both channels play important roles in cell activation, migration, and proliferation through the regulation of membrane potential and calcium signalling. We hypothesize that K(Ca)3.1- and/or K(v)1.3 dependent cell processes are one of the common denominators in asthma pathophysiology. If true, these channels might serve as novel targets for the treatment of asthma. Emerging evidence lends support to this hypothesis. Further validation through the study of the role that these channels play in normal and asthmatic airway cell (patho)physiology and in vivo models will provide further justification for the assessment of small molecule blockers of K(v)1.3 and K(Ca)3.1 in the treatment of asthma. PMID- 19681867 TI - Curcumin adds spice to the debate: lipid metabolism in liver disease. AB - Activated hepatic stellate cells (HSCs), the major source of the collagens involved in fibrosis and non-alcoholic fatty liver disease (NAFLD), undergo a profound loss of lipid and vitamin A storage capacity, as a consequence of a decline in expression of 'adipogenic' transcription factors such as peroxisome proliferator-activated receptor-gamma (PPARgamma). By contrast, hepatocytes undergo a micro- and macro-vesicular steatosis, reflecting the accumulation of triacylglycerol, and associated with chronic inflammation and fibrosis. These paradoxical findings are extended in this issue: Kang and Chen demonstrate that while low-density lipoproteins (LDL) can activate HSCs, curcumin can inhibit this process by activation of PPARgamma, which not only represses gene expression of SREBP-2 and LDLR, but via induction of expression of SREBP-1c, restores the lipid storage capacity characteristic of quiescent HSCs, suggesting that curcumin may be of therapeutic usage in protecting against liver steatosis and fibrosis. PMID- 19681866 TI - Emerging cardiovascular actions of the incretin hormone glucagon-like peptide-1: potential therapeutic benefits beyond glycaemic control? AB - Glucagon-like peptide-1 (GLP-1) is an incretin hormone secreted by the small intestine in response to nutrient ingestion. It has wide-ranging effects on glucose metabolism, including stimulation of insulin release, inhibition of glucagon secretion, reduction of gastric emptying and augmentation of satiety. Importantly, the insulinotropic actions of GLP-1 are uniquely dependent on ambient glucose concentrations, and it is this particular characteristic which has led to its recent emergence as a treatment for type 2 diabetes. Although the major physiological function of GLP-1 appears to be in relation to glycaemic control, there is growing evidence to suggest that it may also play an important role in the cardiovascular system. GLP-1 receptors (GLP-1Rs) are expressed in the heart and vasculature of both rodents and humans, and recent studies have demonstrated that GLP-1R agonists have wide-ranging cardiovascular actions, such as modulation of heart rate, blood pressure, vascular tone and myocardial contractility. Importantly, it appears that these agents may also have beneficial effects in the setting of cardiovascular disease (CVD). For example, GLP-1 has been found to exert cardioprotective actions in experimental models of dilated cardiomyopathy, hypertensive heart failure and myocardial infarction (MI). Preliminary clinical studies also indicate that GLP-1 infusion may improve cardiac contractile function in chronic heart failure patients with and without diabetes, and in MI patients after successful angioplasty. This review will discuss the current understanding of GLP-1 biology, examine its emerging cardiovascular actions in both health and disease and explore the potential use of GLP-1 as a novel treatment for CVD. PMID- 19681868 TI - Bioavailability of diazepam after intramuscular injection of its water-soluble prodrug alone or with atropine-pralidoxime in healthy volunteers. AB - BACKGROUND AND PURPOSE: The aim of this study was to assess the relative bioavailability of diazepam after administration of diazepam itself or as a water soluble prodrug, avizafone, in humans. EXPERIMENTAL APPROACH: The study was conducted in an open, randomized, single-dose, three-way, cross-over design. Each subject received intramuscular injections of avizafone (20 mg), diazepam (11.3 mg) or avizafone (20 mg) combined with atropine (2 mg) and pralidoxime (350 mg) using a bi-compartmental auto-injector (AIBC). Plasma concentrations of diazepam were quantified using a validated LC/MS-MS assay, and were analysed by both a non compartmental approach and by compartmental modelling. KEY RESULTS: The maximum concentration (C(max)) of diazepam after avizafone injection was higher than that obtained after injection of diazepam itself (231 vs. 148 ng.mL(-1)), while area under the curve (AUC) values were equal. Diazepam concentrations reached their maximal value faster after injection of avizafone. Injection of avizafone with atropine-pralidoxime (AIBC) had no effect on diazepam C(max) and AUC, but the time to C(max) was increased, relative to avizafone injected alone. According to the Akaike criterion, the pharmacokinetics of diazepam after injection as a prodrug was best described as a two-compartment with zero-order absorption model. When atropine and pralidoxime were injected with avizafone, the best pharmacokinetic model was a two-compartment with a first-order absorption model. CONCLUSION AND IMPLICATIONS: Diazepam had a faster entry to the general circulation and achieved higher C(max) after injection of prodrug than after the parent drug. Administration of avizafone in combination with atropine and pralidoxime by AIBC had no significant effect on diazepam AUC and C(max). PMID- 19681869 TI - Flavocoxid, a dual inhibitor of cyclooxygenase and 5-lipoxygenase, blunts pro inflammatory phenotype activation in endotoxin-stimulated macrophages. AB - BACKGROUND AND PURPOSE: The flavonoids, baicalin and catechin, from Scutellaria baicalensis and Acacia catechu, respectively, have been used for various clinical applications. Flavocoxid is a mixed extract containing baicalin and catechin, and acts as a dual inhibitor of cyclooxygenase (COX) and 5-lipoxygenase (LOX) enzymes. The anti-inflammatory activity, measured by protein and gene expression of inflammatory markers, of flavocoxid in rat peritoneal macrophages stimulated with Salmonella enteritidis lipopolysaccharide (LPS) was investigated. EXPERIMENTAL APPROACH: LPS-stimulated (1 microg.mL(-1)) peritoneal rat macrophages were co-incubated with different concentrations of flavocoxid (32-128 microg.mL(-1)) or RPMI medium for different incubation times. Inducible COX-2, 5 LOX, inducible nitric oxide synthase (iNOS) and inhibitory protein kappaB-alpha (IkappaB-alpha) levels were evaluated by Western blot analysis. Nuclear factor kappaB (NF-kappaB) binding activity was investigated by electrophoretic mobility shift assay. Tumour necrosis factor-alpha (TNF-alpha) gene and protein expression were measured by real-time polymerase chain reaction and enzyme-linked immunosorbent assay respectively. Finally, malondialdehyde (MDA) and nitrite levels in macrophage supernatants were evaluated. KEY RESULTS: LPS stimulation induced a pro-inflammatory phenotype in rat peritoneal macrophages. Flavocoxid (128 microg.mL(-1)) significantly inhibited COX-2 (LPS = 18 +/- 2.1; flavocoxid = 3.8 +/- 0.9 integrated intensity), 5-LOX (LPS = 20 +/- 3.8; flavocoxid = 3.1 +/- 0.8 integrated intensity) and iNOS expression (LPS = 15 +/- 1.1; flavocoxid = 4.1 +/- 0.4 integrated intensity), but did not modify COX-1 expression. PGE(2) and LTB(4) levels in culture supernatants were consequently decreased. Flavocoxid also prevented the loss of IkappaB-alpha protein (LPS = 1.9 +/- 0.2; flavocoxid = 7.2 +/- 1.6 integrated intensity), blunted increased NF-kappaB binding activity (LPS = 9.2 +/- 2; flavocoxid = 2.4 +/- 0.7 integrated intensity) and the enhanced TNF-alpha mRNA levels (LPS = 8 +/- 0.9; flavocoxid = 1.9 +/- 0.8 n-fold/beta actin) induced by LPS. Finally, flavocoxid decreased MDA, TNF and nitrite levels from LPS-stimulated macrophages. CONCLUSION AND IMPLICATIONS: Flavocoxid might be useful as a potential anti-inflammatory agent, acting at the level of gene and protein expression. PMID- 19681870 TI - Prevention of arterial stiffening by pyridoxamine in diabetes is associated with inhibition of the pathogenic glycation on aortic collagen. AB - BACKGROUND AND PURPOSE: Our team previously demonstrated that diabetes induces a deterioration in vascular dynamics, in parallel with the enhanced formation of advanced glycation end products. The aim of this study was to determine whether prevention of the arterial stiffening by pyridoxamine in diabetes is associated with inhibition of the pathogenic glycation on aortic collagen. EXPERIMENTAL APPROACH: Diabetes was induced in rats by a single tail vein injection with 55 mg.kg(-1) steptozotocin (STZ). After induction of hyperglycaemia, animals were treated for 8 weeks with pyridoxamine (1 g.L(-1) in drinking water) and compared with the age-matched untreated diabetic controls. Pulse wave reflection along the vasculature was derived using the impulse response function of the filtered aortic input impedance spectra. KEY RESULTS: Treatment of this experimental diabetes with pyridoxamine resulted in a significant increase in wave transit time and a decrease in wave reflection factor, indicating that pyridoxamine attenuates the diabetes-induced augmentation in systolic load of the left ventricle coupled to its arterial system. Meanwhile, pyridoxamine therapy ameliorated the diabetes-related cardiac hypertrophy, as evidenced by the reduction in ratio of the left ventricular weight to body weight. Glycation derived modification of aortic collagen was also found to be attenuated by administration of pyridoxamine to the STZ-induced diabetic rats. CONCLUSIONS AND IMPLICATIONS: Pyridoxamine imparts significant protection against the diabetes induced deterioration in pulsatile arterial load imposed on the heart, at least partly through inhibition of the formation of advanced glycation end products and their accumulation on aortic collagen of the STZ-treated rats. PMID- 19681871 TI - Urethane, but not pentobarbitone, attenuates presynaptic receptor function in rats: a contribution to the choice of anaesthetic. AB - BACKGROUND AND PURPOSE: We examined whether cannabinoid CB(1) and histamine H(3) receptors resemble alpha(2)-adrenoceptors in that their presynaptically mediated cardiovascular effects are less marked in urethane- than in pentobarbitone anaesthetized pithed rats. EXPERIMENTAL APPROACH: Effects of the cannabinoid agonist CP-55,940 and the H(3) receptor agonist imetit on electrically induced tachycardic and vasopressor responses, respectively, was compared in pithed rats anaesthetized with urethane or pentobarbitone. The affinity of urethane for the three receptors was measured by radioligand binding studies in rat brain cortex membranes and its potency assessed in superfused mouse tissues preincubated with (3)H-noradrenaline. KEY RESULTS: The neurogenic tachycardic response was less markedly inhibited by CP-55,940 in urethane- than in pentobarbitone-anaesthetized pithed rats. Imetit inhibited the neurogenic vasopressor response after pentobarbitone but not after urethane. The catecholamine-induced tachycardic and vasopressor response did not differ between rats anaesthetized with either compound. Urethane 10 mM (plasma concentration reached under anaesthesia) did not affect binding to CB(1) or H(3) receptors and alpha(2) adrenoceptors, nor did it alter the inhibitory effect of agonists at the three receptors on electrically evoked (3)H-noradrenaline release. CONCLUSIONS AND IMPLICATIONS: Urethane, but not pentobarbitone, abolished the H(3) receptor-mediated vascular response in pithed rats and attenuated the CB(1) receptor-mediated cardiac response much more than pentobarbitone. The weaker effects of CB(1), H(3) and alpha(2) receptor agonists cannot be explained by antagonism by urethane at the three receptors in vitro. Pentobarbitone, but not urethane, is suitable as an anaesthetic for investigations of inhibitory presynaptic receptor function in pithed and anaesthetized rats. PMID- 19681872 TI - Endogenous cannabinoids induce fever through the activation of CB1 receptors. AB - BACKGROUND AND PURPOSE: The effects of centrally administered cannabinoids on body core temperature (Tc) and the contribution of endogenous cannabinoids to thermoregulation and fever induced by lipopolysaccharide (LPS) (Sigma Chem. Co., St. Louis, MO, USA) were investigated. EXPERIMENTAL APPROACH: Drug-induced changes in Tc of male Wistar rats were recorded over 6 h using a thermistor probe (Yellow Springs Instruments 402, Dayton, OH, USA) inserted into the rectum. KEY RESULTS: Injection of anandamide [(arachidonoylethanolamide (AEA); Tocris, Ellisville, MO, USA], 0.01-1 microg i.c.v. or 0.1-100 ng intra-hypothalamic (i.h.), induced graded increases in Tc (peaks 1.5 and 1.6 degrees C at 4 h after 1 microg i.c.v. or 10 ng i.h.). The effect of AEA (1 microg, i.c.v.) was preceded by decreases in tail skin temperature and heat loss index (values at 1.5 h: vehicle 0.62, AEA 0.48). Bell-shaped curves were obtained for the increase in Tc induced by the fatty acid amide hydrolase inhibitor [3-(3-carbamoylphenyl)phenyl] N-cyclohexylcarbamate (Cayman Chemical Co., Ann Arbor, MI, USA) (0.001-1 ng i.c.v.; peak 1.9 degrees C at 5 h after 0.1 ng) and arachidonyl-2 chloroethylamide (ACEA; Tocris) (selective CB(1) agonist; 0.001-1 microg i.c.v.; peak 1.4 degrees C 5 h after 0.01 microg), but (R,S)-(+)-(2-Iodo-5-nitrobenzoyl) [1-(1-methyl-piperidin-2-ylmethyl)-1H-indole-3-yl] methanone (Tocris) (selective CB(2) agonist) had no effect on Tc. AEA-induced fever was unaffected by i.c.v. pretreatment with 6-Iodo-2-methyl-1-[2-(4-morpholinyl)ethyl]-1H-indole-3-yl](4 methoxyphenyl) methanone (Tocris) (selective CB(2) antagonist), but reduced by i.c.v. pretreatment with N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4 dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide (AM251; Tocris) (selective CB(1) antagonist). AM251 also reduced the fever induced by ACEA or LPS. CONCLUSIONS AND IMPLICATIONS: The endogenous cannabinoid AEA induces an integrated febrile response through activation of CB(1) receptors. Endocannabinoids participate in the development of the febrile response to LPS constituting a target for antipyretic therapy. PMID- 19681873 TI - Mechanisms involved in the regional haemodynamic effects of intermedin (adrenomedullin 2) compared with adrenomedullin in conscious rats. AB - BACKGROUND AND PURPOSE: Intermedin (IMD) is a newly identified member of the calcitonin family of peptides that shares structural and functional homology with adrenomedullin (AM). In vivo cardiovascular effects of AM have been described, but relatively little is known of the in vivo actions of IMD. The purpose of this study was to compare the regional haemodynamic effects of IMD with those of AM in conscious rats, and investigate possible underlying mechanisms. EXPERIMENTAL APPROACH: Measurements of blood pressure, heart rate and renal, mesenteric and hindquarters haemodynamics were made in conscious, chronically-instrumented rats. KEY RESULTS: IMD caused tachycardia and vasodilatation in all three vascular beds, associated with modest hypotension. At an equimolar dose (1 nmol.kg(-1)), most of the cardiovascular effects of IMD were greater than those of AM. The AM receptor antagonist, AM(22-52), was equally effective in attenuating the renal and mesenteric vasodilator effects of IMD (1 nmol.kg(-1)) and AM (3 nmol.kg(-1)), but inhibition of NO synthase was more effective at reducing the vasodilator effects of IMD than AM. Vascular K(ATP) channel blockade with U-37883A did not inhibit the vasodilator effects of either peptide. CONCLUSIONS AND IMPLICATIONS: In vivo, the regional haemodynamic profile of IMD resembles that of AM, and some of the vasodilator effects of IMD are mediated by AM receptors and NO, but not by K(ATP) channels. The cardiovascular effects of AM have been implicated in various pathological conditions, but whether or not endogenous IMD fulfils a similar role remains to be determined. PMID- 19681875 TI - Quality of life evaluation in epidermolysis bullosa (EB) through the development of the QOLEB questionnaire: an EB-specific quality of life instrument. AB - BACKGROUND: Epidermolysis bullosa (EB) has a profound effect on quality of life (QOL); however, generic QOL assessments are poor indicators of the impact of EB. OBJECTIVES: To develop a valid and reliable EB-specific QOL tool for use in measuring the effects of disease impact and interventions. METHODS: Open, nonstructured interviews were conducted with 26 patients with EB, along with 33 family members and 11 health professionals (70 individuals) for item generation. A pilot questionnaire was compiled, refined and distributed to 130 patients with EB. From the 115 returned questionnaires a principal axis factor analysis was undertaken producing a 17-item final questionnaire. Discriminative validity was assessed by differences in scores between EB subtypes. Content validity was assessed by expert ranking of items in terms of importance. Construct validity was evaluated by correlation with existing QOL tools. Test-retest reliability and internal consistency were evaluated. Factor analysis was performed. RESULTS: A 17 item questionnaire was developed: the QOLEB questionnaire. This gave distinguishing QOL scores to different EB subtypes, and correlated highly with existing QOL instruments. CONCLUSIONS: The QOLEB questionnaire is the first EB specific QOL measurement tool, and is a valid and reliable measurement tool for the quantification of QOL in patients with various subtypes of EB. In addition, the QOLEB has potential as a sensitive instrument to monitor QOL, and to identify dimensions of QOL as targets for interventions and research. PMID- 19681874 TI - Preclinical evaluation of a monoclonal antibody targeting the epidermal growth factor receptor as a radioimmunodiagnostic and radioimmunotherapeutic agent. AB - BACKGROUND AND PURPOSE: The studies described here are the first to evaluate the in vitro and in vivo properties of (111)In-CHX-A''-panitumumab for radioimmunotherapy (alpha- and beta(-)-emitters) and radioimmunoimaging (single photon emission computed tomography and positron emission tomography). EXPERIMENTAL APPROACH: Twenty-seven human carcinoma cell lines were analysed for expression of epidermal growth factor receptors by flow cytometry. Panitumumab was conjugated with CHX-A''-DTPA (diethylenetriamine-pentaacetic acid) and radiolabelled with (111)In. Immunoreactivity of the CHX-A''-DTPA-panitumumab and (111)In-CHX-A''-DTPA-panitumumab was evaluated by radioimmunoassays. Tumour targeting was determined in vivo by direct quantitation of tumour and normal tissues and by gamma-scintigraphy. KEY RESULTS: For 26 of 27 human tumour cell lines, 95% of the cells expressed epidermal growth factor receptors over a range of intensity. Immunoreactivity of panitumumab was retained after modification with CHX-A''-DTPA. Radiolabelling of the immunoconjugate with (111)In was efficient with a specific activity of 19.5 +/- 8.9 mCi.mg(-1) obtained. Immunoreactivity and specificity of binding of the (111)In-panitumumab was shown with A431 cells. Tumour targeting by (111)In-panitumumab was demonstrated in athymic mice bearing A431, HT-29, LS-174T, SHAW or SKOV-3 s.c. xenografts with little uptake observed in normal tissues. The (111)In-panitumumab was also evaluated in non-tumour-bearing mice. Pharmacokinetic studies compared the plasma retention time of the (111)In-panitumumab in both non-tumour-bearing and A431 tumour-bearing mice. Tumour targeting was also visualized by gamma-scintigraphy. CONCLUSIONS AND IMPLICATIONS: Panitumumab can be efficiently radiolabelled with (111)In with high labelling yields. Based on the efficiency in tumour targeting and low normal tissue uptake, panitumumab may be an effective targeting component for radioimmunodiagnostic and radioimmunotherapeutic applications. PMID- 19681876 TI - Hidradenitis suppurativa and inflammatory bowel disease: are they associated? Results of a pilot study. AB - Background The co-occurrence of hidradenitis suppurativa (HS) and Crohn disease (CD) published in a few case reports resulted in the wide acceptance of an association between these two diseases. However, the combined prevalence of these diseases is currently unknown; furthermore, it is unknown whether this co occurrence also applies for ulcerative colitis (UC). Objectives To estimate the prevalence of HS in patients with inflammatory bowel disease (IBD) living in the Southwest of the Netherlands. Methods During an IBD patient information meeting, randomly, 158 patients with IBD were interviewed about recurrent painful boils in the axillae and/or groin and were shown illustrative clinical pictures of the appearance of HS. Results Of the 158 patients interviewed, 102 (65%) had CD and 56 (35%) had UC. Twenty-five people (16%) responded that they had had or still experienced painful boils in the axillae and/or groin, of whom 17 were patients with CD (17%) and eight had UC (14%). Conclusions This pilot study shows for the first time that HS occurs in patients with CD or UC. More prospective studies are warranted to establish the association between HS and IBD and its underlying pathogenesis. PMID- 19681877 TI - Hereditary congenital hypopigmented and hyperpigmented macules (Westerhof syndrome) in two siblings. PMID- 19681878 TI - Immunomodulatory effects of the alpha-melanocyte-stimulating hormone-related tripeptide K(D)PT on human scalp hair follicles under proinflammatory conditions. PMID- 19681879 TI - A comparative study of the effects of disposable and Volkmann spoon curettes in the treatment of basal cell carcinoma. PMID- 19681880 TI - Delay in medical attention to hand eczema: a follow-up study. AB - BACKGROUND: Hand eczema often runs a chronic course but early medical intervention may be assumed to improve the prognosis. OBJECTIVES: To follow patients with hand eczema for 6 months after seeing a dermatologist to investigate if delay in medical attention would impair the prognosis. METHODS: Study participants were 333 patients with hand eczema from nine dermatological clinics in Denmark. Severity of hand eczema was assessed by the patients at baseline and at the 6-month follow up using a self-administered photographic guide. Additional information was obtained by self-administered questionnaires. RESULTS: Median patient delay (defined as the period from onset of symptoms until seeing a general practitioner) was 3 months [interquartile range (IQR) 1.5-8.0]. The median healthcare delay (defined as the period from the first visit to a general practitioner until seeing a dermatologist) was 3 months (IQR 1-8). In a logistic regression model, the odds ratio of a poor prognosis increased by a factor of 1.11 [95% confidence interval (CI) 1.02-1.21] per month of patient delay and by 1.05 (95% CI 1.00-1.10) per month of healthcare delay. CONCLUSIONS: A poorer prognosis of hand eczema was associated with longer delay before medical attention. PMID- 19681881 TI - CONSORT adoption and quality of reporting of randomized controlled trials: a systematic analysis in two dermatology journals. AB - BACKGROUND: CONSORT (Consolidated Standards for Reporting Trials) guidelines were constructed to ensure optimal reporting quality of randomized controlled trials (RCTs). OBJECTIVES: To determine the effect of the adoption of CONSORT on the reporting quality of RCTs, we performed a systematic evaluation of RCTs published in two dermatology journals pre- and post-CONSORT adoption. METHODS: The journals selected for the study were the Journal of the American Academy of Dermatology and the British Journal of Dermatology. We selected RCTs published in 1997 and 2006 using both Medline and hand searching. The following critical CONSORT criteria were recorded: sample size, type of disease studied, type of control, single-centre or multicentre study, type of funding, blinding, methods and type of randomization, definition of a primary endpoint, justification for sample size selection and power calculation, population for analysis, and adequacy of group comparison. A multivariable analysis was conducted to determine factors associated with optimal reporting quality. RESULTS: In total, 98 studies were included. Improvement in reporting quality was evident for the specification of the randomization method (20% in 1997 vs. 45% in 2006, P < 0.01) and for the justification of sample size (22% in 1997 vs. 43% in 2006, P = 0.027). The percentage of studies with optimal reporting quality increased from 11% in 1997 to 28% in 2006 (P = 0.03). Factors significantly associated with a good methodological quality were pharmaceutical industry funding and publication in 2006 vs. 1997. CONCLUSIONS: There is a need to improve the reporting quality of RCTs published in dermatology journals. PMID- 19681882 TI - Systematic comparison of nonmelanoma skin cancer microarray datasets reveals lack of consensus genes. AB - BACKGROUND: DNA microarray technology has revealed vast numbers of gene expression alterations associated with human malignancies. Assigning validity and biological significance to these changes, however, remains a considerable hurdle. Recently, microarray analysis has been applied to the study of nonmelanoma skin cancer. OBJECTIVES: To compare experimental data rigorously in order to strengthen conclusions regarding the pathogenesis of basal cell carcinoma (BCC) and squamous cell carcinoma (SCC), and to evaluate systematically the experimental and statistical parameters that may impact the degree of consensus among differentially expressed genes (DEGs) between studies. METHODS: We performed a systematic comparison of 10 studies that applied DNA microarray technology to study BCC/SCC. RESULTS: A total of 1133 DEGs collectively reported across the studies were compared, and 64 DEG overlaps were found: 18 DEG overlaps in SCC vs. SCC study comparisons, 18 DEG overlaps in BCC vs. BCC study comparisons and 28 DEG overlaps in BCC vs. SCC study comparisons. We documented differences in several experimental methods that may account for the relative lack of consensus between studies, including sample type, tissue procurement/handling, microarray chip and statistical analysis. The two most dysregulated biological pathways across all studies involved genes with enzymatic and structural/adhesion functions. CONCLUSIONS: DEGs that were found to overlap across two or more studies and biological pathways with the largest representation of DEGs across studies may be particularly relevant to disease pathogenesis and serve as targets for future therapy. In future work, more consistent experimental methods across laboratories may improve the validity of reported DEGs and strengthen conclusions drawn from microarray data. PMID- 19681883 TI - Chronic parvovirus B19 infection leading to red cell aplasia following treatment of Hodgkin lymphoma. PMID- 19681884 TI - Iron overload in thalassaemia intermedia: reassessment of iron chelation strategies. AB - Thalassaemia intermedia (TI) is a syndrome marked by its diverse underlying genetic basis although its pathophysiology remains unclear, particularly regarding the nature of iron loading and toxicity. It is, however, evident that there are key differences from the extensively studied thalassaemia major (TM) population and caution is required when assessing iron load based on serum ferritin values, as this approach is known to underestimate the true extent of iron loading in patients with TI. Although effective iron chelation therapy has been available for many years, studies in TI-specific populations are rare and evidence suggests that management of iron levels may be less rigorous than in patients with TM and other chronic anaemias. Better understanding of the need to assess and treat iron overload in both transfused and non-transfused TI patients is clearly required. PMID- 19681885 TI - Elevation of clonal serum free light chains in patients with HIV-negative primary effusion lymphoma (PEL) and PEL-like lymphoma. PMID- 19681886 TI - Stem cell donation--what advice can be given to the donor? AB - Haematopoietic stem cell transplantation (HSCT) is widely used to treat patients with a range of haematological and non-haematological disorders. Both bone marrow and peripheral blood stem cell collection are associated with morbidity and, very rarely, mortality. We investigated the information that exists to adequately inform donors about the relative merits of each procedure. We carried out a systematic review analysing data from six prospective randomised controlled trials of related donors and discuss here the merits and drawbacks of this approach. Registry data mostly describes patient outcome but stem cell donor registries collect and report information on unrelated donors which could easily be extended to related donors. Further well-designed, randomised studies are required. PMID- 19681887 TI - MicroRNA expression in chronic lymphocytic leukaemia. PMID- 19681888 TI - Reintoxication: the release of fat-stored delta(9)-tetrahydrocannabinol (THC) into blood is enhanced by food deprivation or ACTH exposure. AB - BACKGROUND AND PURPOSE: Delta(9)-tetrahydrocannabinol (THC), the main psychoactive constituent of cannabis, accumulates in adipose tissue where it is stored for long periods of time. Here we investigated whether conditions that promote lipolysis can liberate THC from adipocytes to yield increased blood levels of THC. EXPERIMENTAL APPROACH: In vitro studies involved freshly isolated rat adipocytes that were incubated with THC before exposure to the lipolytic agent adrenocorticotrophic hormone (ACTH). A complementary in vivo approach examined the effects of both food deprivation and ACTH on blood levels of THC in rats that had been repeatedly injected with THC (10 mg.kg(-1)) for 10 consecutive days. Lipolysis promoted by ACTH or food deprivation was indexed by measurement of glycerol levels. KEY RESULTS: ACTH increased THC levels in the medium of THC pretreated adipocytes in vitro. ACTH also enhanced THC release from adipocytes in vitro when taken from rats repeatedly pretreated with THC in vivo. Finally, in vivo ACTH exposure and 24 h food deprivation both enhanced the levels of THC and its metabolite, (-)-11-nor-9-carboxy-Delta(9)-tetrahydrocannabinol (THC-COOH) in the blood of rats that had been pre-exposed to repeated THC injections. CONCLUSIONS AND IMPLICATIONS: The present study shows that lipolysis enhances the release of THC from fat stores back into blood. This suggests the likelihood of 'reintoxication' whereby food deprivation or stress may raise blood THC levels in animals chronically exposed to the drug. Further research will need to confirm whether this can lead to functional effects, such as impaired cognitive function or 'flashbacks'. PMID- 19681889 TI - Inhibition of the JNK signalling pathway enhances proteasome inhibitor-induced apoptosis of kidney cancer cells by suppression of BAG3 expression. AB - BACKGROUND AND PURPOSE: Proteasome inhibitors represent a novel class of anti tumour agents that have clinical efficacy against haematological and solid cancers. The anti-apoptotic protein BAG3 is a member of the Bcl-2-associated athanogene family. We have previously shown that BAG3 is up-regulated after exposure to proteasome inhibitors and that inhibition of BAG3 sensitized cells to apoptosis induced by proteasome inhibition. However, the mechanisms by which proteasome inhibition induced BAG3 expression remained unclear and the present experiments were designed to elucidate these mechanisms. EXPERIMENTAL APPROACH: Effects of the proteasome inhibitor MG132 on activation of mitogenic signalling pathways were evaluated in kidney cancer cells (A498, Caki1, Caki2), with Western blotting. Specific inhibitors against individual mitogenic signalling pathways, real-time reverse transcription-polymerase chain reaction and luciferase reporter assays were used to investigate the roles of mitogenic signalling pathways in BAG3 induction after proteasome inhibition. Cell death was evaluated using Annexin V/propidium iodide staining and subsequent FACS. KEY RESULTS: MG132 activated several key mitogenic signalling pathways including extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 mitogen activated protein kinase (MAPK) activities. Induction of BAG3 by MG132 was inhibited by blocking JNK, but not ERK1/2 and p38 MAPK signalling pathways. In addition, SP600125 and dominant-negative JNK1 suppressed BAG3 promoter-driven reporter gene expression. Furthermore, activation of the JNK pathway induced BAG in kidney cancer cells after treatment with MG132. CONCLUSIONS AND IMPLICATIONS: Our results suggested that the JNK pathway was associated with the protective response against proteasome inhibition, by mediating induction of BAG3. PMID- 19681890 TI - Robert F. Furchgott, Nobel laureate (1916-2009)--a personal reflection. AB - Robert F. Furchgott, pharmacologist and joint winner of the Nobel Prize for Medicine or Physiology (1998) died on the 12th of May 2009 aged 92. By unlocking the astonishingly diverse biological actions of nitric oxide, Furchgott leaves behind a rich legacy that has both revolutionized our understanding of human physiology and stimulated new and exciting opportunities for drug development in a wide range of pathological conditions. In this article, William Martin, who worked with Furchgott for 2 years (1983-1985), following the exciting discovery of endothelium-derived relaxing factor/nitric oxide, pays tribute to his close friend and colleague. PMID- 19681891 TI - Capsular involvement in patients undergoing partial nephrectomy for localized renal cell carcinoma: an adverse pathological finding? AB - OBJECTIVE: To evaluate the prognostic impact of capsular involvement (CaI) in patients treated exclusively with partial nephrectomy (PN) for localized renal cell carcinoma (RCC), as in these patients CaI was recently reported as an adverse prognostic factor. PATIENTS AND METHODS: We retrospectively reviewed the medical records of patients treated with PN for a sporadic and localized RCC (pT1 pT2N0M0) in our institution between 1985 and 2005. Univariate and multivariate analysis using a Cox proportional-hazards regression analysis were conducted to identify significant predictors of oncological outcome for several clinical and pathological factors, i.e. imperative indication, histological type, Fuhrman grade, tumour size, T stage, CaI, and surgical margins. Disease-free and specific survival rates of patients with CaI and no evidence of CaI were compared using the log-rank test. RESULTS: In all, 305 patients had a PN for localized RCC, of whom 22 (7.2%) had CaI in the PN specimen. The median (range) follow-up was 6 (1.5-23) years. Multivariate statistical analysis showed that imperative indication for PN and high-grade RCC were independently associated with worse disease-free and -specific survival, whereas CaI had no prognostic value. Disease free and -specific survival in patients with and without CaI were not significantly different at 5 and 10 years. CONCLUSIONS: In a contemporary series of patients exclusively treated with PN for localized RCC, CaI was not predictive of disease recurrence and disease-specific mortality. These results do not support the use of any change in postoperative management in patients with CaI after PN. PMID- 19681892 TI - Long-term outcome of ureterosigmoidostomy: an analysis of patients with >10 years of follow-up. AB - STUDY TYPE: Therapy (case series). LEVEL OF EVIDENCE: 4. OBJECTIVE: To examine our long-term experience with ureterosigmoidostomy (USS) to evaluate its potential applicability in the treatment of benign and malignant conditions of the urinary bladder, as USS has been largely disregarded recently, secondary to concerns of long-term complications, but has had a resurgence of interest due to its potential applicability to newer minimally invasive surgical techniques. PATIENTS AND METHODS: We identified 51 patients who had USS from 1956 to 2006 at our institution and with >10 years of follow-up. The patients were followed retrospectively by a chart review. Patient data were analysed in a multifaceted fashion, paying particular attention to metabolic abnormalities, early (< or =30 days) and late (>30 days) complication rates, continence rates, imaging changes, and the rate of repeat surgical intervention. RESULTS: The median (range) follow up was 15.7 (10.0-45.4) years and the median age at surgery was 58.8 (0.4-79.0) years; 40 (79%) patients had the procedure for malignancy and 11 (22%) for benign disease. Six patients (12%) had at least one early complication, including one wound dehiscence and one pulmonary embolus. In all, 22 patients (43%) had at least one late complication, with anastomotic stricture being the most common (11/51, 22%). This was followed by recurrent pyelonephritis in eight patients (16%), stones in five (10%), chronic renal insufficiency in three (6%) and severe intractable acidosis in two (4%). A repeat surgical intervention was required in 19 (37%) patients. In all, 94% (48) reported complete continence. No patient developed colonic malignancy during the course of this study. CONCLUSIONS: USS is associated with long-term complications. While this complication rate might not be acceptable for all patients, some might be willing to undergo the procedure as the primary method of urinary diversion. When designing newer minimally invasive techniques for the treatment of benign and malignant conditions of the bladder, consideration could be given to USS as a form of urinary diversion in highly selected patients. PMID- 19681893 TI - Sympathetic skin response in patients with erectile dysfunction. AB - OBJECTIVES: To evaluate the role of the sympathetic skin response (SSR) in men with erectile dysfunction (ED), focusing on detecting SSR in the penis. PATIENTS AND METHODS: We assessed the SSR in 82 patients with ED, as an indicator of abnormalities both in amyelinic C-fibres and in autonomic pathways in these patients. The SSR was carried out according to the to the Technical Standards of the International Federation of Clinical Neurophysiology. Electrical stimulation was applied through superficial electrodes over the contralateral median nerve. Values were recorded with superficial electrodes on the skin in the contralateral hand and foot, as well as in the penis. The percentage of SSR (SSR%) was classified into three groups, i.e. 0-20%, 21-89% and 90-100%. Results of latency were also classified into three groups of normal or abnormal (increased) latency, and response blocking (no response), the last two being considered pathological conditions. RESULTS: In the penis, the mean (sd) SSR% was 52.8 (43.19)% and significantly lower than responses in hands and feet. There was a significant correlation of the SSR% between the palm of the hand and the sole of the foot (P = 0.01) and between the sole of foot and penis (P = 0.05). Diabetics showed a significant decrease (P = 0.001) in the mean SSR% in the palm of the hand and sole of the foot. Although not statistically different, the mean SSR% in the penis was lower in diabetics than in patients with other risk factors for ED. Likewise, the mean SSR% in hand, foot and penis increased with an increase in the International Index of Erectile Function. In the penis, latency was normal (<1.5 ms) in 14 and abnormal in 37 patients. There was a significant association between pathological chronic re-innervation in the bulbocavernosus muscle and SSR latencies in the foot (P = 0.002) and penis (P = 0.03). Bulbocavernosus muscle electromyography showed a higher frequency of chronic bilateral axonomnesis in patients with abnormal latencies (28%) than in patients with normal SSR latencies in the penis. CONCLUSION: These results establish an indication of the SSR in patients with ED, registering responses not only in classic locations like the palm of the hand or sole of the foot, but also in the penis. The SSR% was useful as an indicator of the effect on efferent C fibres. Despite SSR being a polysynaptic potential of long latency and regulated by the cerebral cortex, the present results show that it is advisable to record the latencies of SSR in the three areas registered, and especially in the penis, where it seems be more useful as a marker of lumbosacral and/or pudendal alterations. PMID- 19681894 TI - Inhibition of bladder tumour growth by histone deacetylase inhibitor. AB - OBJECTIVE: To examine the expression profile of histone deacetylase (HDAC)-1 and explore its potential role in the development of bladder cancer, using valproic acid (VPA), a HDAC inhibitor, which reduces tumour growth and metastasis formation in animal models. MATERIALS AND METHODS: The study comprised clinical samples from patients with urinary bladder cancer, mouse urinary bladder tissue specimens, and two human urinary bladder cancer cell lines (HT-1376 and 5637). HDAC1 mRNA and protein expression were examined using real-time reverse transcription-polymerase chain reaction and immunohistochemical methods. Female C3H/He mice were given VPA (0, 250, 500 and 750 mg/kg body weight, intraperitoneal, every day) from the start or 4 weeks after 0.05%N-butyl-N-(4 hydroxybutyl)-nitrosamine (BBN) treatment, and were humanely killed and sampled at 8 and 12 weeks. RESULTS: A significantly higher level of HDAC1 mRNA was expressed in human urinary bladder cancer specimens. The immunohistochemical study showed that HDAC1 was expressed in the cytoplasm and nucleus in the specimens. BBN treatment increased HDAC1 mRNA expression in the urinary bladder. VPA administration seemed to delay the incidences of BBN-induced mouse urinary bladder tumour, possibly through p21(WAF1) protein expression. CONCLUSION: These results indicate that HDAC might be an effective molecular target for cancer therapy. PMID- 19681895 TI - Immediate surgical outcomes for radical prostatectomy in the University HealthSystem Consortium Clinical Data Base: the impact of hospital case volume, hospital size and geographical region on 48,000 patients. AB - OBJECTIVE: To determine the impact of hospital variables on immediate surgical outcomes for patients treated with radical prostatectomy (RP) in academic centres. PATIENTS AND METHODS: The University HealthSystem Consortium (UHC) Clinical Data Base was queried for data corresponding to patients who had RP at one of 130 academic medical centres nationwide between 2003 and the second quarter of 2007 (48,086). RP case volume (1-99, 100-499 and >500), total discharges (1-49,999, 50,000-99,999 >100,000), and geographical region (five categories) were determined and categorized for each academic centre. Analysis of variance and the Tukey statistic were used to assess the results. Length of stay (LOS), intensive care unit (ICU) rate, complication rate (CR) and in-hospital mortality (IHM) were analysed. RESULTS: Case volume was a significant predictor of LOS, ICU and CR. The mean LOS was 3.77, 2.65 and 2.09 days, respectively, for centres from three tiers of lowest to highest case volumes (P < 0.001). ICU rates for the three tiers were 18.57, 3.61, and 1.30 (P < 0.001); CRs were 15.93, 8.79 and 5.76 (P < 0.001). Tukey analysis showed a 'ceiling' effect for ICU and CRs; there were no differences between the two higher case-volume groups. IHM was not significantly different between groups stratified by case volume. Stratification by total discharges showed differences in ICU rates only (P = 0.003). Stratification by geographical region showed no differences in outcome. CONCLUSIONS: RP case volume was an important variable in predicting three of the four outcome variables. CRs and ICU rates showed a 'ceiling effect' suggesting that an unknown 'critical volume' of cases portends improved surgical outcomes. PMID- 19681896 TI - Dual segmental arterial supply to a single renal lobule: a study using semi serial sections of mid-term fetuses. AB - OBJECTIVE: To determine which and how many segmental arteries supply a single renal lobule especially at and near the segmental border, as no data are available on the arterial supply to a single renal lobule (a pyramid and its covering cortex) at the border between renal segments. MATERIALS AND METHODS: We studied semi-serial sections (100 microm intervals; haematoxylin and eosin staining) of 26 kidneys obtained from 13 mid-term human fetuses (20-30 weeks gestation). Eleven fetuses were used for horizontal sections and two for sagittal sections. Section to section, we first traced the renal pyramids using a pen to understand the lobular configuration. RESULTS: Fusion of the medullary pyramids was usually evident along the segmental borders, except for the border between the upper and middle segments. This fusion often accompanied a pair of arcuate arteries, each of which arose from a different renal segment, but shared a common corticomedullary junction to supply a single lobule. The incidence of fusion was high along the anterior border of the posterior segment as well as the border between the apical and upper segments. When the lower segmental artery entered the parenchyma distant from the renal hilus, fusion of pyramids occurred along the border between the posterior and lower segments in association with double arcuate arteries. CONCLUSIONS: A single lobule is most likely to be shared by double segments at or near the segmental border. This morphology seems to be a result of lobule fusion during development and growth. Thus, in adults, a dual segmental supply to a single lobule seems to be more frequent than in fetuses. PMID- 19681897 TI - Factors associated with low and high voiding frequency in children with diurnal urinary incontinence. AB - STUDY TYPE: Prognosis (cohort). LEVEL OF EVIDENCE: 2a. OBJECTIVE: To investigate, in a population-based study, the association of urinary voiding frequency and daytime wetting or diurnal urinary incontinence (UI) in children aged 6.5 years, to test the hypothesis that children with high or low voiding frequency have increased somatic and psychological risks. SUBJECTS AND METHODS: Participants were a cohort of > 8000 children enrolled in the population-based Avon Longitudinal Study of Parents and Children, a prospective longitudinal study of an original birth cohort of nearly 14,000 children. Parents completed postal questionnaires asking about diurnal UI, voiding frequency and associated somatic and behavioural signs and symptoms. RESULTS: In all, 10.4% of children had diurnal UI. Children with diurnal UI had significantly more gastrointestinal, urinary and psychological symptoms than the controls. Of the children with diurnal UI, 35.7% had a voiding frequency of < 5 voids/day, 61.5% 5-9 voids/day and 2.8% > or = 10 voids/day. High voiding frequency was more common in boys, children with frequent diurnal UI and those with combined diurnal and night-time UI, and also in children with nocturia, urgency, hyperactivity and conduct problems. Children with a low voiding frequency needed to be reminded to go to the toilet more often. Fecal incontinence, constipation, stomach 'aches', 'choosy' eating and urinary tract infections were not associated with voiding frequency. CONCLUSIONS: Voiding frequency is an important symptom in children with diurnal UI. The children with high voiding frequencies are especially at risk for associated somatic and behavioural problems. PMID- 19681898 TI - A nomogram for predicting upgrading in patients with low- and intermediate-grade prostate cancer in the era of extended prostate sampling. AB - STUDY TYPE: Diagnostic (exploratory cohort). LEVEL OF EVIDENCE: 2b. OBJECTIVE: To develop a nomogram to predict the probability that the pathological Gleason sum (GS) will be higher than that indicated by the biopsy, suggesting a higher risk for the patient presumed to be at low risk, as a substantial proportion of patients with low and intermediate grade on biopsy are upgraded on interpretation of the radical prostatectomy (RP) specimens, but a similar clarification of accurate Gleason scoring is not available in patients with no surgical histology. PATIENTS AND METHODS: The study included 1017 patients who had RP after biopsy showing GS 6 and 7 (3 + 4) from 2000 to 2007. Nomogram predictor variables included age, race, digital rectal examination, prostate-specific antigen (PSA) level, number of cores taken, number of positive cores, maximum percentage cancer in any core, number of previous biopsies, prostate volume, clinical stage, high grade prostatic intraepithelial neoplasia, atypical small acinar proliferation, inflammation and perineural invasion. We calculated the nomogram-predicted probability in each patient. The area under the receiver operating characteristic curve was calculated as a measure of discrimination, and the calibration was assessed graphically. RESULTS: The mean age of the patients was 60 years, the mean PSA level 6.62 ng/mL; 336 patients were upgraded (33%), 623 remained the same (61.3%) and 58 were downgraded (5.7%). A nomogram for predicting the possibility of upgrading was constructed that had a concordance index of 0.68. The nomogram was well calibrated. CONCLUSIONS: Our nomogram for predicting upgrading provides important additional information for deciding on treatment to both the urologist and the patient with low- and intermediate-grade prostate cancer. It might prove useful when the possibility of a more aggressive Gleason variant can change the management, and is especially meaningful when management options other than surgery are selected based on the inability to recognize the true pathological actual GS. PMID- 19681899 TI - The second-line management of idiopathic overactive bladder: what is the place of sacral neuromodulation and botulinum toxin-A in contemporary practice? PMID- 19681900 TI - Renal functional outcomes for tumours in a solitary kidney managed by ablative or extirpative techniques. AB - OBJECTIVE: To present the renal functional outcomes for patients treated with open partial nephrectomy (OPN) or radiofrequency ablation (RFA) for tumours in a solitary kidney, as renal masses in a solitary kidney present a challenging treatment dilemma. PATIENTS AND METHODS: A retrospective review of institutional databases identified 89 patients with 98 renal tumours in a solitary kidney managed by RFA or OPN under cold ischaemia between January 1997 and September 2007. The choice of therapy was based on patient and surgeon preference, tumour characteristics and comorbidities. Renal function was calculated using the modified Modification of Diet in Renal Disease equation. RESULTS: Outcomes from 47 patients treated by RFA and 42 by OPN were analysed at a median follow-up of 18.1 and 30.0 months, respectively (P = 0.02). The median age (65.9 vs 59.6 years, P = 0.03) and American Society of Anesthesiology score (3.0 vs 2.0, P = 0.01) were both higher in patients treated with RFA. The median tumour size was greater for tumours managed by OPN (3.9 vs 2.8 cm, P = 0.001), while the median preoperative glomerular filtration rate (GFR) was lower in the RFA group (46.5 vs 55.9 mL/min/1.73 m(2) for OPN, P = 0.04). Compared to RFA, patients treated with OPN had a greater decline in GFR at all times evaluated, including soon after the procedure (15.8% vs 7.1%), 12 months after surgery (24.5% vs 10.4%) and at the last follow-up (28.6% vs 11.4%, all P < 0.001). For patients with a pretreatment GFR of > 60 or > 30 mL/min/1.73 m(2), there was a new onset of decline in GFR of <60 and <30 mL/min/1.73 m(2) in none and 7% of patients after RFA, and in 35% and 17% after OPN. CONCLUSION: Ablative techniques, which obviate ischaemic insults, might be a particularly attractive option for managing tumours in solitary renal units at risk of declining function. Renal functional outcomes compare favourably to extirpative surgery using cold ischaemia. PMID- 19681901 TI - The independent value of tumour volume in a contemporary cohort of men treated with radical prostatectomy for clinically localized disease. AB - OBJECTIVE: To determine if prostate tumour volume is an independent prognostic factor in a contemporary cohort of men who had a radical prostatectomy (RP) for clinically localized disease, as the effect of tumour volume on prostate cancer outcomes has not been consistently shown in the era of widespread screening with prostate-specific antigen (PSA). PATIENTS AND METHODS: The study included 856 men who had RP from 1998 to 2007 for localized prostate cancer. Tumour volume based on pathology was analysed as a continuous and categorized (<0.26, 0.26-0.50, 0.51 1.00, 1.01-2.00, 2.01-4.00, >4.00 mL) variable using Cox proportional hazards regression and Kaplan-Meier analysis. A multivariable analysis was also conducted controlling for PSA level, Gleason grade, surgical margins, and pathological stage. RESULTS: Tumour volume had a positive association with grade and stage, but did not correlate with biochemical recurrence-free survival on univariate analysis as a continuous variable (hazard ratio 1.00, P = 0.09), and was only statistically significant for volumes of >4 mL as a categorical variable. No tumour volume was an independent predictor of prostate cancer recurrence on multivariate analysis. There was no difference between tumour volume and time to cancer recurrence for organ-confined tumours using Kaplan-Meier analysis. In low risk patients (PSA level <10 ng/mL, Gleason score < or = 6, clinical stage T1c/T2a) tumour volume did not correlate with biochemical recurrence-free survival in univariate or multivariable analysis. CONCLUSIONS: There is no evidence that tumour volume is an independent predictor of prostate cancer outcome and it should not be considered as a marker of tumour risk, behaviour or prognosis. PMID- 19681902 TI - Evaluation of the serum testosterone to prostate-specific antigen ratio as a predictor of prostate cancer risk. AB - OBJECTIVE: To analyse the ratio of serum testosterone (sT) to prostate-specific antigen (PSA) as a predictor of prostate cancer risk, as low levels of sT have been related to a greater risk of prostate cancer, and its ratio with serum PSA level was recently proposed as a new tool to increase the specificity of PSA. PATIENTS AND METHODS: In all, 439 consecutive men with a normal digital rectal examination and a serum PSA level of 4.1-20 ng/mL had a transrectal ultrasonography-guided biopsy using a 10-core scheme, with an additional 1-8 cores according to prostate volume and patient age. The sT level was determined before the procedure using a chemiluminescent assay, and the ratio of sT to PSA (sT/PSA) was calculated after transforming sT measurements from ng/dL to ng/mL. The percentage free PSA (%fPSA) and PSA density were also included in this analysis. RESULTS: The overall cancer detection rate was 42.1%. The median sT level was 469 ng/dL in men with cancer and 499 ng/dL in those without (P = 0.521). The median sT/PSA was 0.68 and 0.74, respectively (P = 0.215). However, the median %fPSA was 14 in men with cancer and 17 in men without (P < 0.001) and the median PSA density was 0.22 and 0.16, respectively (P < 0.001). The multivariate analysis confirmed the independent predictive value only for %fPSA (odds ratio 0.94, 95% confidence interval 0.91-0.98) and PSA density (5.8, 3.42 19.8). CONCLUSION: These results do not support the use of sT/PSA for predicting the risk of prostate cancer and to increase the specificity of PSA. PMID- 19681903 TI - Expression of podoplanin, CD44, and p63 in squamous cell carcinoma of the lung. AB - Recent molecular biological studies have identified podoplanin as a candidate cancer stem cell (CSC) marker in squamous cell carcinoma (SqCC). The purpose of this study was to examine the expression pattern of podoplanin, and the other stem cell markers CD44 and p63, and their relationship to clinico-pathological features including survival in pulmonary SqCC. We examined histologically the expression of podoplanin, CD44, and p63 in 162 consecutive SqCC by immunostaining. Podoplanin expression was observed in 107 (66%) tumors, CD44 in 145 (89.5%), and p63 in 151 (93.2%), respectively. In 95.3% of the podoplanin positive tumors, tumor cells showing strong expression were localized in the periphery of the tumor nests. However, this peripheral localization was observed in only 55.9% of the CD44-positive and 43% of p63-positive tumors. In 88.8% of the podoplanin-positive tumors, positive cells were localized more peripherally in the tumor nests than CD44- or p63-positive cells and when CD44 and p63 expressions were compared in these podoplanin-positive tumors, p63-positive layers in the periphery of the tumor nests were broader compared to CD44-positive layers. These findings suggest tumor cells are aligned in the "hierarchical distribution pattern" according to the expression of these three markers. Patients who had podoplanin-positive tumors with the "hierarchical pattern" resulted in significantly better overall survival than those who had podoplanin negative tumors (P = 0.043). These results suggest that podoplanin expression would reflect the most immature status in the differentiation process of SqCC, and SqCC with hierarchical expression would be a well-organized tumor group with lower biological aggressiveness based on the CSC concept. PMID- 19681904 TI - Involvement of Pin1 induction in epithelial-mesenchymal transition of tamoxifen resistant breast cancer cells. AB - Acquisition of resistance to tamoxifen is a critical therapeutic problem in breast cancer patients. Epithelial-mesenchymal transition (EMT), where cells undergo a developmental switch from a polarized epithelial phenotype to a highly motile mesenchymal phenotype, is associated with invasion and motility of cancer cells. Here, we found that tamoxifen-resistant (TAMR)-MCF-7 cells had undergone EMT, as evidenced by mesenchymal-like cell shape, downregulation of basal E cadherin expression, and overexpression of N-cadherin and vimentin, as well as increased Snail transcriptional activity and protein expression. Given the roles of glycogen synthase kinase (GSK)-3beta and nuclear factor (NF)-kappaB in Snail mediated E-cadherin deregulation during EMT, we examined the role of these signaling pathways in the EMT of TAMR-MCF-7 cells. Both Ser9-phosphorylated GSK 3beta (inactive form) and NF-kappaB reporter activity were increased in TAMR-MCF 7 cells, as was activation of the phosphatase and tensin homolog depleted on chromosome ten (PTEN)-phosphoinositide 3 (PI3)-kinase-Akt pathway. Pin1, a peptidyl-prolyl isomerase, was overexpressed in TAMR-MCF-7 cells, and Snail transcription and the expression of EMT markers could be decreased by Pin1 siRNA treatment. These results imply that Pin1 overexpression in TAMR-MCF-7 cells is involved in the EMT process via PTEN-PI3-kinase-Akt-GSK-3beta and/or GSK-3beta-NF kappaB-dependent Snail activation, and suggest the potential involvement of Pin1 in EMT during breast cancer development. PMID- 19681905 TI - Cellular characterization of the peritumoral edema zone in malignant brain tumors. AB - Brain edema is a hallmark of human malignant brain tumors and contributes to the clinical course and outcome of brain tumor patients. The so-called perifocal edema or brain swelling imposes in T2-weighted MR scans as high intensity areas surrounding the bulk tumor mass. The mechanisms of this increased fluid attraction and the cellular composition of the microenvironment are only partially understood. In this study, we focus on imaging perifocal edema in orthotopically implanted gliomas in rodents and correlate perifocal edema with immunohistochemical markers. We identified that areas of perifocal edema not only include the tumor invasion zone, but also are associated with increased glial fibrillary acidic protein (GFAP) and aquaporin-4 expression surrounding the bulk tumor mass. Moreover, a high number of activated microglial cells expressing CD11b and macrophage migration inhibitory factor (MIF) accumulate at the tumor border. Thus, the area of perifocal edema is mainly dominated by reactive changes of vital brain tissue. These data corroborate that perifocal edema identified in T2-weighted MR scans are characterized with alterations in glial cell distribution and marker expression forming an inflammatory tumor microenvironment. PMID- 19681906 TI - Internalization-dependent recognition of Mycobacterium avium ssp. paratuberculosis by intestinal epithelial cells. AB - Mycobacterium avium ssp. paratuberculosis (MAP) is the causative agent of Johne's disease, a highly prevalent chronic intestinal infection in domestic and wildlife ruminants. The microbial pathogenesis of MAP infection has attracted additional attention due to an association with the human enteric inflammatory Crohn's disease. MAP is acquired by the faecal-oral route prompting us to study the interaction with differentiated intestinal epithelial cells. MAP was rapidly internalized and accumulated in a late endosomal compartment. In contrast to other opportunistic mycobacteria or M. bovis, MAP induced significant epithelial activation as indicated by a NF-kappaB-independent but Erk-dependent chemokine secretion. Surprisingly, MAP-induced chemokine production was completely internalization-dependent as inhibition of Rac-dependent bacterial uptake abolished epithelial activation. In accordance, innate immune recognition of MAP by differentiated intestinal epithelial cells occurred through the intracellularly localized pattern recognition receptors toll-like receptor 9 and NOD1 with signal transduction via the adaptor molecules MyD88 and RIP2. The internalization-dependent innate immune activation of intestinal epithelial cells is in contrast to the stimulation of professional phagocytes by extracellular bacterial constituents and might significantly contribute to the histopathological changes observed during enteric MAP infection. PMID- 19681907 TI - Cell invasion of Yersinia pseudotuberculosis by invasin and YadA requires protein kinase C, phospholipase C-gamma1 and Akt kinase. AB - The outer membrane proteins YadA and invasin of Yersinia pseudotuberculosis promote invasion into mammalian cells through beta(1)-integrins and trigger the production of interleukin (IL)-8. FAK, c-Src and the PI3 kinase were previously found to be important for both YadA- and invasin-promoted uptake. Here, we demonstrate that two different downstream effectors of PI3 kinase, Akt and phospholipase Cgamma1 are required for efficient cell invasion. Inhibition of Akt or phospholipase C-gamma (PLC-gamma)1 by pharmaceutical agents as well as reduced expression of the isoforms Akt1 and Akt2, and of PLC-gamma1 by RNA interference decreased entry of YadA- and Inv-expressing bacteria significantly. In addition, we report that the conventional protein kinases C (PKC)alpha and -beta, positioned downstream of PLC-gamma1, are activated upon Inv- or YadA-promoted cell entry. They colocalize with intracellular bacteria and their depletion by siRNA treatment also resulted in a strong reduction of cell entry. In contrast, neither Akt nor PLC-gamma1, and the PKCs are essential for YadA- and Inv-mediated IL-8 synthesis and release. We conclude that YadA and invasin of Y. pseudotuberculosis both trigger similar signal transduction pathways during integrin-mediated phagocytosis into epithelial cells, which lead to the activation of Akt, PLC-gamma1, PKCalpha and -beta downstream of PI3 kinase, separate from the MAPK-dependent pathway that triggers IL-8 production. PMID- 19681908 TI - N-glycosylated proteins and distinct lipooligosaccharide glycoforms of Campylobacter jejuni target the human C-type lectin receptor MGL. AB - An increasing number of bacterial pathogens produce an array of glycoproteins of unknown function. Here we report that Campylobacter jejuni proteins that are modified by the N-linked glycosylation machinery encoded by the pgl locus bind the human Macrophage Galactose-type lectin (MGL). MGL receptor binding was abrogated by EDTA and N-acetylgalactosamine (GalNAc) and was successfully transferred to Escherichia coli by introducing the C. jejuni pgl locus together with a glycan acceptor protein. In addition to glycoproteins, C. jejuni lipooligosaccharide with a terminal GalNAc residue was recognized by MGL. Recombinant E. coli expressing the C. jejuni pgl locus in the absence of a suitable glycan acceptor protein produced altered lipopolysaccharide glycoforms that gained MGL reactivity. Infection assays demonstrated high levels of GalNAc dependent interaction of the recombinant E. coli with MGL-transfected mammalian cells. In addition, interleukin-6 production by human dendritic cells was enhanced by C. jejuni lacking N-linked glycans compared with wild-type bacteria. Collectively, our results provide evidence that both N-linked glycoproteins and distinct lipooligosaccharide glycoforms of C. jejuni are ligands for the human C type lectin MGL and that the C. jejuni N-glycosylation machinery can be exploited to target recombinant bacteria to MGL-expressing eukaryotic cells. PMID- 19681909 TI - Cell type-specific effects of Yersinia pseudotuberculosis virulence effectors. AB - One important feature of Yersinia pseudotuberculosis that enables resistance against the host immune defence is delivery of the antiphagocytic effectors YopH and YopE into phagocytic cells. The tyrosine phosphatase YopH influences integrin signalling, and YopE impairs cytoskeletal dynamics by inactivating Rho GTPases. Here, we report the impact of these effectors on internalization by dendritic cells (DCs), which internalize antigens to orchestrate host immune responses. We found that this pathogen resists internalization by DCs via YopE. YopH that is important for blocking phagocytosis by macrophages and neutrophils and which is also present inside the DCs does not contribute to the resistance. However, the YopH targets Fyb and p130Cas show higher expression levels in macrophages than in DCs. Furthermore, live cell microscopy revealed that the cells internalize Y. pseudotuberculosis in different ways: the macrophages utilize a locally restricted receptor-mediated zipper mechanism, whereas DCs utilize macropinocytosis involving constitutive ruffling that randomly catches bacteria into membrane folds. We conclude that YopH impacts early phagocytic signalling from the integrin receptor to which the bacterium binds and that this tight receptor-mediated stimulation is absent in DC macropinocytosis. Inactivation of cytoskeletal dynamics by YopE affects ruffling activity and hence also internalization. The different modes of internalization can be coupled to the major functions of these respective cell types: elimination by phagocytosis and antigen sampling. PMID- 19681912 TI - Increased oxidized low density lipoprotein associated with high ceruloplasmin activity in patients with active acromegaly. AB - OBJECTIVE: Active acromegaly is associated with increased mortality from cardiovascular causes. Several studies have shown increased atherogenic risk factors and biomarkers of inflammation and atherosclerosis in association with growth hormone excess. The aim of this study was to evaluate oxidized low density lipoprotein (oxLDL) levels and some modulators of LDL oxidative modification in patients with acromegaly. DESIGN: Open transversal study. PATIENTS: Fifteen patients with active acromegaly and 15 controls were studied. MEASUREMENTS: We evaluated the levels of oxLDL, thiobarbituric acid reactive substances (TBARS), ceruloplasmin, bilirubin, uric acid and total reactive antioxidant potential, and the activities of ceruloplasmin, myeloperoxidase, superoxide distmutase, paraoxonase 1, and platelet activating factor acethylhydrolase. Statistical analysis was performed including body mass index as a covariate or as a fixed variable. RESULTS: Patients with acromegaly showed significantly higher levels of oxLDL (120 +/- 19 vs. 86 +/- 20 U/l, P < 0.001) and endothelin (P < 0.05), increased ceruloplasmin activity (P < 0.01) and a trend towards higher values in TBARS concentration (P = 0.07) in comparison to healthy controls. OxLDL was positively associated with GH, IGF-I and its binding protein 3 (r = 0.63, P < 0.001; r = 0.53, P < 0.01; and r = 0.56, P < 0.01; respectively). OxLDL showed direct associations with endothelin-1 (r = 0.53, P < 0.01) and ceruloplasmin activity (r = 0.43, P < 0.05). The other parameters evaluated were similar in both groups. CONCLUSIONS: The increase in plasma oxLDL levels, a direct marker of the plaque formation, could constitute a link between atherosclerosis and active acromegaly. LDL oxidation would not be the consequence of diminished antioxidant defences, but of an enhancement in prooxidant factors like ceruloplasmin. PMID- 19681913 TI - Polymorphisms of the UCP2 gene are associated with proliferative diabetic retinopathy in patients with diabetes mellitus. AB - BACKGROUND AND OBJECTIVE: Uncoupling protein 2 (UCP2) plays a role in controlling reactive oxygen species (ROS) production by mitochondria. As ROS overproduction is related to diabetic retinopathy (DR), UCP2 gene polymorphisms might be involved in the development of this complication. We investigated whether the 866G/A (rs659366), Ala55Val (rs660339) and 45 bp insertion/deletion (Ins/Del) polymorphisms in the UCP2 gene might be associated with proliferative DR (PDR). DESIGN AND METHODS: In this case-control study, we analysed 501 type 2 diabetic patients (242 patients with PDR and 259 subjects without any degree of DR) and 196 type 1 diabetic patients (85 cases with PDR and 111 without DR). Haplotypes constructed from the combination of the three UCP2 polymorphisms were inferred using a Bayesian statistical method. RESULTS: In the type 2 diabetic group, multivariate analyses confirmed that the haplotype [A Val Ins] was an independent risk factor for PDR when present in one [adjusted odds ratio (aOR) = 2.12; P = 0.006], at least one (aOR = 2.75; P = 0.00001), or two copies (aOR = 5.30; P = 0.00001), suggesting an additive model of inheritance. Nevertheless, in type 1 diabetic patients, the association of this haplotype with PDR was confirmed only when it was present in at least one (aOR = 2.68; P = 0.014) or two copies (aOR = 6.02; P = 0.005). CONCLUSIONS: The haplotype [A Val Ins] seems to be an important risk factor associated with PDR in both type 2 and 1 diabetic groups. PMID- 19681914 TI - Dexamethasone administration inhibits skeletal muscle expression of the androgen receptor and IGF-1--implications for steroid-induced myopathy. AB - CONTEXT: Glucocorticoids are a well-recognized cause of muscle weakness. The early effects of glucocorticoids on skeletal muscle (SkM) androgen and IGF-1 pathways have not been previously investigated in human subjects. OBJECTIVE: To determine if administration of the potent glucocorticoid dexamethasone down regulates SkM androgen receptor and the IGF-1 signalling pathway. METHODS AND SUBJECTS: Twenty-four subjects (12 men and 12 women), including 12 with type 2 diabetes and 12 nondiabetics were enrolled. Venous blood sampling and biopsy of vastus lateralis were performed before and after administration of oral dexamethasone 4 mg/day for 4 days. MAIN OUTCOME MEASURES: Changes in plasma testosterone and IGF-1, SkM androgen receptor mRNA, SkM IGF-1mRNA and SkM IGF-1 receptor mRNA by quantitative RT-PCR after dexamethasone. RESULTS: Relative expression of SkM androgen receptor was similar in male (1.63 +/- 0.37) vs. female (1.57 +/- 0.30) subjects, despite the significant difference in plasma testosterone levels. Plasma IGF-1 and SkM expression of IGF-1 and IGF-1 receptor were also similar between males and females. Following dexamethasone, there was a significant down-regulation of SkM androgen receptor (1.60 +/- 0.23 vs. 1.11 +/- 0.16, P < 0.05) and IGF-1 (1.72 +/- 0.29 vs. 1.06 +/- 0.14, P < 0.05) mRNA, but no change in expression of the IGF-1 receptor. Plasma testosterone fell significantly in both sexes (male: 15.0 +/- 1.3 vs. 11.3 +/- 1.2 nmol/l, P < 0.01, female: 1.8 +/- 0.5 vs. 0.5 +/- 0.1 nmol/l, P < 0.05). CONCLUSIONS: Exogenous steroid excess results in relative androgen deficiency at two levels, reduced circulating testosterone and SkM androgen receptor mRNA, along with reduced SkM IGF-1 mRNA. These defects may contribute to the development of steroid-induced myopathy. PMID- 19681915 TI - Severe hyperthyroidism: aetiology, clinical features and treatment outcome. AB - BACKGROUND: Severe hyperthyroidism (SH) is a serious medical disorder that can compromise life. There have not been systematic studies in which SH has been evaluated in detail. Here, our aims were: (1) to analyse both clinical and analytical features and outcome in patients with SH and (2) to compare these data with those found in more usual forms of hyperthyroidism. Patients and methods All patients diagnosed of SH (free thyroxine, FT4 > 100 pmol/l, NR: 11-23) seen in our endocrinology clinic in the last 15 years were studied and compared with a sample of patients with mild (mH; FT4, 23-50 pmol/l) and moderate (MH; FT4, 51 100 pmol/l) hyperthyroidism. Aetiology, clinical analytical and imaging data at diagnosis, therapeutic response and outcome were registered. Results A total of 107 patients with overt hyperthyroidism (81 females, mean age +/- SD 46.9 +/- 16.1 years) were evaluated. We studied a historic group with SH (n = 21; 14 females, 40.9 +/- 17.2 years) and, as a comparator group, we analyszed the data of 86 hyperthyroid patients (67 females, 48.4 +/- 15.5.6 years, NS) comparable in age and gender. The comparator group was classified in MH (n = 37, 26 females, 47.2 +/- 16.6 years) and mH (n = 49, 41 females, 49.4 +/- 14.8 years). In comparison with mH group, SH patients were significantly (P < 0.05) younger and showed a greater proportion of first episode of thyroid hyperfunction (P < 0.05). Graves' disease was the main aetiology in the three groups, but patients with SH showed the highest titre of TSH-receptor antibodies (TRAb) (P < 0.001). Heart rate and size of goitre were higher in SH group than in mH and MH groups (P < 0.01). Atrial fibrillation was more frequently reported in SH group than in MH and mH groups (15.8%vs. 5.4% and 0%, respectively, P < 0.05). RESULTS: from logistic regression analysis showed that younger age [OR 0.958 (95% CI, 0.923 0.995), P = 0.026], presence of asthenia [OR 4.35 (1.48-12.78), P = 0.008] and higher heart rate [OR 1.03 (1.01-1.06), P = 0.013] were independent clinical variables associated to SH. SH patients showed similar biochemical parameters in comparison with mH group, except for increased serum aspartate aminotransferase (AST) (P < 0.01) and calcium (P < 0.05) levels, and decreased serum cholesterol (P < 0.05) and albumin (P < 0.05) concentrations. Logistic regression analysis showed that only AST [OR 1.07 (1.02-1.11), P = 0.005] was an independent biochemical variable associated to SH. No differences in the type of therapy, cure rate and time in achieving cure were found in SH subjects in relation to patients with milder forms of hyperthyroidism. FT4 was the only independent predictor of cure [OR 0.98 (CI 95%, 0.97-0.99), P < 0.05]. CONCLUSIONS: Graves' disease is the most common aetiology in patients with SH. This type of hyperthyroidism is usually de novo and is accompanied by more clinical signs, symptoms, and analytical derangements, as well as higher titres of TRAb at diagnosis than milder forms of hyperthyroidism. The present data are not able to show differences in treatment modality, time to achieve cure, and remission rate among patients with mild, moderate and severe hyperthyroidism. PMID- 19681916 TI - The common exon 3 polymorphism of the growth hormone receptor gene and the effect of growth hormone therapy on growth in Korean patients with Turner syndrome. AB - OBJECTIVE: Recombinant human growth hormone (GH) can achieve final adult height gain in girls with Turner syndrome (TS), but its efficacy varies widely across individuals. The exon 3-deleted polymorphism of growth hormone receptor (d3-GHR) has been reported to be associated with responsiveness to GH therapy. The short term growth response of Turner patients to GH therapy was analysed according to their GHR-exon 3 polymorphism genotype. DESIGN AND PATIENTS: This was a retrospective study of 175 TS patients. Auxological and endocrine parameters were measured, and the GHR-exon 3 genotype was analysed. Allelic frequencies of GHR exon 3 genotype were compared between patients with TS and control individuals. GH had been administered to 147 patients, 115 of which remained pre-pubertal after the first follow-up year. Changes in height standard deviation score (SDS), height velocity (HV), body mass index (BMI), IGF-1 and IGF binding protein-3 (IGFBP-3) concentrations were compared between these patients, grouped according to genotype, after the first follow-up year. RESULTS: There was no difference in GHR-exon 3 genotype frequency between the TS and control groups of Koreans. According to the GHR-exon 3 genotype (fl/fl group vs. d3/fl and d3/d3 group), HV gain and height SDS gain did not differ significantly at the first year of GH therapy. Moreover, changes in IGF-1, IGFBP-3 concentration and BMI showed no significant difference between the groups with and without d3-GHR after 1 year of GH therapy. CONCLUSION: The distribution of the GHR-exon 3 genotype was similar in the TS and control groups in a Korean population. The growth promotion efficacy of GH therapy did not differ significantly between TS patients with and without the d3-GHR allele. These findings indicate that the GHR-exon 3 genotype may not be a major factor to affect the GH response in Korean Turner patients. PMID- 19681917 TI - The role of genetic variation in peroxisome proliferator-activated receptors in the polycystic ovary syndrome (PCOS): an original case-control study followed by systematic review and meta-analysis of existing evidence. AB - OBJECTIVE: To study the association of polymorphisms in the genes encoding peroxisome proliferator-activated receptors (PPARs) with the polycystic ovary syndrome (PCOS). DESIGN: Case-control study and meta-analysis of published evidence. PATIENTS: One hundred and sixty-one polycystic ovary syndrome patients and 113 non-hyperandrogenic women. MEASUREMENTS: Genotyping for PPAR-gamma coactivator-1 gene (PPARGC1A) Gly482Ser, PPAR-alpha Leu162Val, PPAR-delta rs2267668A/G, PPAR-delta-87T/C, PPAR-gamma2 Pro12Ala and PPAR-gamma2 -681C/G variants and systematic review of the literature using the Entrez-PubMed search engine, followed by meta-analysis whenever possible. RESULTS: Polycystic ovary syndrome patients carried the Gly482Ser variant in PPARGC1A more frequently than controls (72%vs. 58%, chi(2 )=( )5.54 P = 0.019), whereas carriers of the PPAR alpha Leu162Val, PPAR-delta rs2267668A/G, PPAR-delta-87T/C, PPAR-gamma2 Pro12Ala and PPAR-gamma2 -681C/G variants were distributed similarly among both groups. The interaction between the PPARGC1A Gly482Ser and PPAR-delta-87T/C variants was also associated with PCOS (OR = 1.24, 95% CI 1.05-1.50, P = 0.008). The systematic review identified 31 studies addressing associations between PPARs variants and PCOS; meta-analysis was possible for nine studies focusing on the PPAR-gamma2 Pro12Ala variant. Although the individual studies did not reveal any statistically significant association, meta-analysis uncovered that carrying the PPAR-gamma2 Pro12Ala variant was associated with a reduced probability of having PCOS (OR = 0.77, 95% CI 0.61-0.96, P = 0.025), and that this association may be mediated by an effect on insulin sensitivity. CONCLUSIONS: Common polymorphisms in the PPARGC1A, PPAR-delta and PPAR-gamma2 loci are associated with PCOS. PMID- 19681918 TI - Atorvastatin pretreatment augments the effect of metformin in patients with polycystic ovary syndrome (PCOS). PMID- 19681919 TI - The autologous serum skin test may be used as a marker for histamine releasing autoantibodies in urticaria and is not relevant to other subject groups. PMID- 19681920 TI - Fixed drug eruption caused by pamabrom. PMID- 19681921 TI - Associations between decay-accelerating factor polymorphisms and allergic respiratory diseases. AB - BACKGROUND: Allergic diseases such as asthma and allergic rhinitis are major causes of morbidity in developed countries. The pathology underlying allergic respiratory diseases is considered to be IgE-mediated type I allergy characterized by mucosal inflammation that occurs in response to allergen exposure. They are common diseases involving a complex inheritance. Complement systems are known to play an important role in allergic diseases. Decay accelerating factor (DAF) is important for the regulation of the complement system and is a good candidate for determining the susceptibility to allergic diseases. OBJECTIVE: The present study aimed to investigate whether polymorphisms in the DAF gene are associated with allergic respiratory diseases in the Japanese population. METHODS: We performed mutation screenings of DAF and conducted a tag single-nucleotide polymorphisms (SNP) association analysis for 684 unrelated adult individuals with seasonal allergic rhinitis (SAR) with Japanese ceder pollen, 188 mite-sensitive adults with asthma, and 346 unrelated non-allergic healthy controls. RESULTS: DAF is located in the tight linkage disequilibrium (LD) block spanning 62 kb. The tag SNP analysis revealed that rs10746463 was significantly associated with SAR (P=0.00033) and mite-sensitive adult asthma (P=0.044). The rs2564978 and rs3841376 haplotypes, which are located in the promoter region of DAF, were in complete LD with rs10746463 (r2=1). Luciferase reporter assays with constructs containing the 5' flanking regions of DAF showed that the plasmid with rs2564978 C/rs3841376 deletion (the risk haplotype) had a statistically significantly lower transcriptional activity than that containing the rs2564978 T/rs3841376 insertion. CONCLUSIONS: Our results suggest that DAF is one of the genes involved in conferring susceptibility to allergic respiratory diseases and show that decreased levels of DAF may be associated with the enhanced specific IgE responses occurring in allergic diseases in the Japanese population. PMID- 19681922 TI - Comparison of horizontal corneal diameter measurements using Galilei, EyeSys and Orbscan II systems. AB - BACKGROUND: White-to-white corneal measurement is used frequently in cataract and refractive surgery as well as in diagnosing various corneal diseases and monitoring congenital glaucoma. In this article, we determine agreement in horizontal corneal diameter measurements in normal eyes among the Galilei V4.01, the EyeSys and the Orbscan IIz. METHODS: In this prospective study, 37 consecutive volunteers were enrolled. All 74 eyes were measured with all three devices. Galilei, EyeSys and Orbscan II measurements were compared with one another. In addition, the data from the right and left eyes were compared for each device. RESULTS: The mean horizontal corneal diameter measurements with the Galilei, the EyeSys and the Orbscan II were 12.01 +/- 0.61 mm, 12.09 +/- 0.87 mm and 11.67 +/- 0.29 mm, respectively. The difference between the Galilei and EyeSys systems was not statistically significant (p = 0.76), however, the Orbscan II showed smaller measurements (p < 0.001). The best 95% limits of agreement between devices were for the Galilei and the Orbscan II (-0.72, 1.48; r = 0.4, p < 0.01). The best 95% limits of agreement between two eyes for each device were found with the Orbscan II (-0.15, 0.17; r = 0.996, p < 0.01). There was no correlation between measurements of right and left eyes for the Galilei or the EyeSys (r = 0.274, p = 0.176 and r = 0.31, p = 0.065, respectively). CONCLUSIONS: These results suggest that measurements made with the Orbscan II are smaller than those obtained with the EyeSys Corneal Analysis system and the Galilei. Among the three devices, the Galilei and the Orbscan II showed the best agreement, however, it is inadvisable to use the three devices interchangeably in every clinical situation. PMID- 19681923 TI - Patients' expectations before and evaluation after dental implant therapy. AB - BACKGROUND: Patients' expectations may influence their satisfaction with implant treatment, but there is a paucity of reports comparing patient expectations and their posttreatment satisfaction. PURPOSE: The aim was to compare patients' expectations before with their satisfaction regarding function and esthetics after dental implant therapy. MATERIALS AND METHODS: Fifty volunteers (49 years +/- 11.45) answered a questionnaire about influencing variables and underwent an initial examination followed by implant therapy. Their expectations regarding esthetics and function were verified on a visual analogue scale (VAS) before treatment. The VAS was also used for posttreatment completion rating. RESULTS: The average ratings for esthetic and functional expectations were 5.0 +/- 2.6 and 5.0 +/- 3.0; the posttreatment completion ratings were 9.1 +/- 1.1 and 9.0 +/- 1.7, respectively. The posttreatment completion ratings significantly exceeded expectations (p < .001, Wilcoxon test). Positive correlations were found between expectations and posttreatment completion ratings for esthetics (Spearman's rho = 0.496, p < .001) and function (Spearman's rho = 0.706, p < .001). An inverse correlation was found between age and functional expectations (Spearman's rho = 0.313, p = .027). CONCLUSIONS: Patients' posttreatment completion ratings significantly exceeded their initial expectations. Expectations and posttreatment completion ratings were irrespective of smoking habits, location in the jaw, sex, or educational level. PMID- 19681924 TI - Evaluation of different methods of indirect sinus floor elevation for elevation heights of 10mm: an experimental ex vivo study. AB - OBJECTIVE: The aim of this study was to macroscopically and microscopically evaluate different methods of indirect sinus floor elevation regarding elevation heights of 10 mm. MATERIALS AND METHODS: Four different methods of indirect sinus floor elevation-osteotome sinus floor elevation (OSFE), bone added osteotome sinus floor elevation (BAOSFE), piezo- surgical sinus floor elevation (PSFE), and sinus floor elevation with an inflatable balloon, balloon-lift-control system (BLC) - were macroscopically and microscopically investigated ex vivo using 36 bisected pigs' heads. RESULTS: OSFE and BAOSFE perforated the Schneiderian membrane, whereas the inflatable balloon caused no laceration. PSFE elevated the mucosa without laceration as well, but was technically restricted to an elevation height of 5 mm. BAOSFE, PSFE, and BLC separated the mucosa, leaving the periosteum on the bone. OSFE completely lifted the soft tissue from the bone, including the periosteum. CONCLUSIONS: The results of this study indicate that balloon elevation of the sinus floor may extend the indication for indirect sinus floor elevation for elevation heights of up to 10 mm. The histological elevation layer seems to be non-uniform in the different sinus floor elevation methods. Further in vivo experiments have to prove these findings as well as their relevance regarding the clinical outcome of sinus floor augmentation. PMID- 19681925 TI - Dimensional tolerances and assembly accuracy of dental implants and machined versus cast-on abutments. AB - BACKGROUND: The clinical application of prosthetic components obtained by different manufacturing processes lacks technological foundation: the dimensional tolerance of individual parts and their assembly accuracy are not known. The rotational misfit (RM) of the hexagonal connection is critical in single-tooth implant restorations, but no standard control procedures are available for its evaluation. PURPOSE: The research aimed at proposing a new protocol for the dimensional assessment of implant-abutment connections, based on noncontact measurement and statistical data processing. The procedure was applied to machined- and cast-on abutments, as well of the matching implants. MATERIALS AND METHODS: Three groups of five abutments each were studied: machined titanium abutments, pre-machined calcinable abutments before casting procedures and the same specimens after casting. A group of five corresponding implants was considered as well. Twice the apothem was measured on each hexagon through an optical measuring microscope. The data were processed to obtain the international tolerance (IT) grade. The RM was then calculated using the apothems of the external and the internal hexagon. RESULTS: All the components were classified between IT8 and IT9, and the maximum RM was around 3-4 degrees for all the assemblies, inferior to the critical limits for the screw joint stability. CONCLUSION: An original measuring protocol was developed, independent of parts assembly and based on ITs. An objective dimensional characterization of prosthetic components and assemblies has been achieved, which is the basis for their reliability in clinical applications. PMID- 19681926 TI - Implants (3.3 mm diameter) for the rehabilitation of edentulous posterior regions: a retrospective clinical study with up to 11 years of follow-up. AB - BACKGROUND: There is limited evidence for the use of narrow-diameter implants for rehabilitation of the posterior regions of the jaws using different surgical techniques. PURPOSE: The purpose of this study was to report the clinical results of implant-supported prosthetic rehabilitations in the posterior regions of both jaws, using narrow-diameter implants. MATERIALS AND METHODS: The study included 147 patients (115 males and 32 females), with an age range of 26 to 77 years (mean = 47.5 years), with a total of 247 implants inserted and followed between 1 and 11 years, with a median follow-up time of 5 years. The patients were in need of fixed prosthetic implant-supported rehabilitations in the posterior region of the jaw, presenting a reduced interradicular bone or a thin alveolar crest. The implant survival estimate was computed using the Kaplan-Meier product limit estimator. RESULTS: The survival rate for narrow diameter implants was 95.1% at 11 years (Kaplan-Meier), with a distribution of 91.4% at 11 years, 95.9% at 10 years, and 95.5% at 9 years for the two-stage, one-stage, and immediate function techniques, respectively. The mean marginal bone resorption recorded at 1, 5, and 10 years were 1.16, 1.53, and 1.74 mm, respectively. Backward conditional logistic regression identified "type of implant" as a strong protective factor against implant failure (MkIII and NobelSpeedy implants compared to the MkII implant; OR = 0.14), and "type of rehabilitation" as a strong risk factor for implant failure (partial rehabilitations compared to single teeth rehabilitations; OR = 4.75). CONCLUSIONS: The results indicate that within the limitations of this study, the use of narrow-diameter implants for the prosthetic rehabilitation of posterior regions of the jaws is viable, with good outcomes in the long-term, irrespective of the surgical technique implemented. PMID- 19681927 TI - Impact fracture resistance of two titanium-abutment systems versus a single-piece ceramic implant. AB - BACKGROUND: The number of patients with oral implants has increased significantly. However, the literature addressing the effect of impact force on titanium and/or ceramic implants is inconclusive. This study sought to determine the fracture resistance to impact load of titanium and ceramic endosseous oral implants. MATERIALS AND METHODS: Endosseous oral implants were vertically positioned in two different mounting media: brass and a bone-simulation material. The implant configurations tested included an experimental one-piece Y-TZP implant and a commercially available titanium implant (external hex) with both titanium and zirconia abutments. The specimens were subjected to an impact load using a pendulum impact tester with tup weights varying from 0.9 to 4.5 kg delivered at a radius of 40.64 mm. Loads were delivered to the abutment at a point 4.27 mm above the implant fixture and block junction. Statistical differences (p < .05) were established using the F-test for variances and, when different, t-test assuming unequal variances. RESULTS: For implants clamped in brass, the titanium implant with titanium abutment required the greatest energy to fracture the implant-abutment system (only the abutment screw failed). The ceramic implant and ceramic abutment on titanium implant presented the lowest fracture energy (p < .01). No significant differences were observed when different systems were inserted into the foam blocks of the bone substitute (p > .25). CONCLUSION: This investigation showed that the fracture energy of two titanium-abutment systems versus a single-piece Y-TZP implant in foam blocks simulating bone elastic modulus was not different, and that differences occurred when the embedding material elastic modulus was increased an order of magnitude. PMID- 19681928 TI - Thin bioactive ceramic-coated alumina-blasted/acid-etched implant surface enhances biomechanical fixation of implants: an experimental study in dogs. AB - BACKGROUND: Thin bioceramic coatings have been regarded as potential substitutes for plasma-sprayed hydroxyapatite coatings. PURPOSE: This study tested the hypothesis that a thin bioactive ceramic coating deposition on an alumina blasted/acid-etched (AB/AE) surface would positively affect the biomechanical fixation and bone-to-implant contact (BIC) of plateau root form implants. MATERIALS AND METHODS: Implants of two different lengths (i.e., 4.5 * 11 mm long, n = 36) and 4.5 * 6 mm (short, n = 36) and two different surfaces, that is, control (AB/AE) and test (AB/AE + 300 - 500 nm bioactive ceramic coating), were placed in the proximal tibiae of six beagle dogs. The implants were retrieved for analyses 2 and 4 weeks after placement. The implants in bone specimens were subjected to torque loads until a 10% drop of the maximum torque was recorded. The specimens were evaluated under optical microscopy for bone morphology and percent BIC. Statistical analysis was performed by a generalized linear mixed effects analysis of variance model and statistical significance set at p < 0.05. RESULTS: Significantly higher torque-to-interface fracture levels for test surface groups of both lengths when compared to control surfaces were observed. No significant difference in BIC was observed between test and control implants of equal length. Histomorphological analysis showed higher degrees of bone organization between the plateaus of test implant surfaces at both implantation times. CONCLUSION: Because the presence of a thin bioactive ceramic coating on the surface did not affect BIC, but positively affected implant biomechanical fixation, the hypothesis was partially validated. PMID- 19681929 TI - Primary stability of a hybrid self-tapping implant compared to a cylindrical non self-tapping implant with respect to drilling protocols in an ex vivo model. AB - BACKGROUND: Modifications of implant design have been intending to improve primary stability. However, little is known about investigation of a hybrid self tapping implant on primary stability. PURPOSES: The aims of this study were to evaluate the primary stability of two hybrid self-tapping implants compared to one cylindrical non-self-tapping implant, and to elucidate the relevance of drilling protocols on primary stability in an ex vivo model. MATERIALS AND METHODS: Two types of hybrid self-tapping implants (Straumann(r) Bone Level implant [BL], Straumann(r) Tapered Effect implant [TE]) and one type of cylindrical non-self-tapping implant (Straumann(r) Standard Plus implant [SP]) were investigated in the study. In porcine iliac cancellous bones, 10 implants each were inserted either using standard drilling or under-dimensioned drilling protocol. The evaluation of implant-bone interface stability was carried out by records of maximum insertion torque, the Periotest(r) (Siemens, Bensheim, Germany), the resonance frequency analysis (RFA), and the push-out test. RESULTS: In each drilling group, the maximum insertion torque values of BL and TE were significantly higher than SP (p=.014 and p=.047, respectively). In each group, the Periotest values of TE were significantly lower than SP (p=.036 and p=.033, respectively). The Periotest values of BL and TE were significantly lower in the group of under-dimensioned drilling than standard drilling (p=.002 and p=.02, respectively). In the RFA, no statistical significances were found in implants between two groups and between implants in each group. In each group, the push out values of BL and TE were significantly higher than SP (p=.006 and p=.049, respectively). CONCLUSION: Hybrid self-tapping implants could achieve a high primary stability which predicts them for use in low-density bone. However, there is still a debate to clarify the influence of under-dimensioned drilling on primary stability. PMID- 19681930 TI - Healing of extraction sockets filled with BoneCeramic(r) prior to implant placement: preliminary histological findings. AB - BACKGROUND: Various grafting materials have been designed to minimize edentulous ridge volume loss following tooth extraction by encouraging new bone formation in healing sockets. BoneCeramic(r) is a composite of hydroxyapatite and beta tricalcium phosphate with pores of 100-500 microns. PURPOSES: The aim of this study was to evaluate bone regeneration in healing sockets substituted with BoneCeramic(r) prior to implant procedures. MATERIALS AND METHODS: Fifteen extraction sockets were substituted with BoneCeramic(r) and 14 sockets were left to heal naturally in 10 patients (mean age 59.6 years). Biopsies were collected only from the implant recipient sites during surgery after healing periods ranging from 6-74 weeks (mean 22). In total, 24 biopsies were available; 10 from substituted and 14 from naturally healed sites. In one site, the implant was not placed intentionally and, in four substituted sites, implant placement had to be postponed due to inappropriate healing, hence from five sites biopsies were not available. Histological sections were examined by transmitted light microscope. RESULTS: At the time of implant surgery, bone at substituted sites was softer than in controls, compromising initial implant stability. New bone formation at substituted sites was consistently poorer than in controls, presenting predominantly loose connective tissue and less woven bone. CONCLUSION: The use of BoneCeramic(r) as a grafting material in fresh extraction sockets appears to interfere with normal healing processes of the alveolar bone. On the basis of the present preliminary findings, its indication as a material for bone augmentation, when implant placement is considered within 6-38 weeks after extraction, should be revised. PMID- 19681931 TI - Regenerative treatment of peri-implantitis using bone substitutes and membrane: a systematic review. AB - PURPOSE: This systematic review aimed to assess the available literature for regenerative treatment of peri-implantitis using bone graft substitutes and membranes. METHODS: A search in electronic databases was conducted to assess all types of clinical studies treating bone defects derived from peri-implantitis using guided bone regeneration (GBR) techniques. RESULTS: During the first screening, 399 titles were identified. Finally, 17 articles reporting on 173 implants were included. The articles mainly focused on radiographic bone fill of the defect. Qualitative measures of "bone fill" were reported: 10.4% of the implants showed complete "bone fill," whereas 85.5% revealed incomplete defect closure. No bone fill was shown in 4.0%. Little information (in 53.2%) was provided regarding the probing depth before or after treatment. Data concerning the inflammatory status of soft tissues were also scarce and only reported in three studies. A large heterogeneity concerning disinfection protocols and regenerative materials used was found. The high percentage of low-quality studies rendered a meta-analysis impossible. CONCLUSION: Complete fill of the bony defect using GBR seems not to be a predictable outcome. The mucosal health status is left unconsidered in most studies. Well-controlled trials are needed to determine predictable treatment protocols for the successful regenerative treatment of peri implantitis using GBR technique. PMID- 19681933 TI - Osteonecrosis of the posterior mandible after implant insertion: a clinical and histological case report. AB - BACKGROUND: In the past few years, the occurrence of an oral lesion, called osteonecrosis of the jaw (ONJ), has been increasingly reported in patients undergoing treatment with bisphosphonates (BPs); however, few published histological studies of ONJ can be found in the literature. PURPOSE: The aim of the present case was to report an occurrence of ONJ after implant insertion. MATERIALS AND METHODS: Multiple myeloma was diagnosed to a 65-year-old female. After 5 years of treatment with intravenous clodronate, two dental implants were inserted in the mandibular molar region. No preexisting bone lesions were present at a preoperative panoramic radiography. Before implant insertion, the patient had suspended the treatment with clodronate for 3 months. Four months after the implant insertion, a breakdown of the oral mucosa covering the implants occurred with a purulent discharge; periapical radiolucency was present around both implants. An en-block resection on the alveolar bone including the two implants was performed. No signs of recurrence of the lesion were observed after a follow up of 20 months. RESULTS: At the interface of one of the implants, a gap was observed between bone and implant. This bone was nonvital, and many osteocyte lacunae were empty. Moreover, this bone appeared to be partially demineralised. No newly formed bone or osteoblasts were present. Bone trabeculae were observed, on the other hand, within the apical implant threads of the other implant. A close connection was observed between this bone and the implant surface. DISCUSSION: The histological findings showed some areas with osseointegration in patients undergoing BP treatment for malignant disease; however, any invasive procedure can determine the onset of osteonecrosis. CONCLUSION: In conclusion, there is certainly a temporal association between BP use and development of ONJ, but a correlation does not necessarily mean causation. Moreover, generalizations about this complex relationship cannot be made on the basis of a single case report. In patients undergoing intravenous treatment, clinicians must be aware of the increased risk of implant failure and, probably, implant insertion should be avoided at all until more conclusive data are available. PMID- 19681932 TI - Marginal bone level changes at dental implants after 5 years in function: a meta analysis. AB - BACKGROUND: It is important that peri-implant bone breakdown caused by, for example, undue load and/or peri-implantitis, is prevented or minimized. Some continuous loss of marginal bone is generally accepted, but the question remains as to what extent it must occur. PURPOSE: The purpose of this study was to compile and compare data on peri-implant marginal bone level changes from prospective studies that have registered the peri-implant marginal bone level radiographically at the time of prosthetic loading, and after 5 years of follow up for implant systems currently available on the market. MATERIALS AND METHODS: A literature search was carried out to identify prospective studies on peri implant marginal bone level changes around dental implants. To be included in a meta-analysis, the implant systems should have been subjected to at least two independent studies. Copycats without documentation were not accepted. RESULTS: Forty prospective studies that presented with a 5-year data were identified. Three implant systems met the inclusion criteria of having at least two independent studies; Astra Tech Dental Implant System(r) (Astra Tech AB, Molndal, Sweden), Branemark System (Nobel Biocare AB, Goteborg, Sweden), and Straumann Dental Implant System (Institute Straumann AG, Basel, Switzerland). The pooled mean marginal bone level change amounted to -0.24 mm (95% CI -0.345, -0.135) for the Astra Tech Dental Implant System, 0.75 mm (95% CI -0.802, -0.693) for the Branemark System, and 0.48 mm (95% CI -0.598, -0.360) for the Straumann Dental Implant System over 5 years, with a statistically significant difference (p < .01) between the systems. CONCLUSIONS: The identified implant systems showed an annual bone loss below or much below what hitherto has been set up as a limit for success. A careful documentation of marginal bone level changes should be mandatory for all implant systems before being marketed. It is also time for revision of existing success criteria to refine the basis for clinical quality judgment of implant treatment. PMID- 19681934 TI - Implant-supported electrostimulating device to treat xerostomia: a preliminary study. AB - BACKGROUND: The full accomplishment of salivary function depends on proper salivary flow rate and composition. Salivary secretion is highly essential in the maintenance of health and integrity of oral hard and soft tissue. Xerostomia is a common symptom affecting between one-fifth and one-third of the adult population, more commonly women than men. Induction of salivary secretion exists in several pharmacological formulations per os. Electrostimulation to enhance salivary secretion has been used frequently as a research tool but only in limited extent as a clinical method to treat patients with xerostomia. PURPOSE: The aims of this preliminary study were to observe and evaluate the therapeutic effect on xerostomia of the Saliwell Crown (Saliwell Ltd., Harutzim, Israel), an innovative saliva electrostimulation device fixed on an implant, placed in the lower third molar area. MATERIALS AND METHODS: A Saliwell Crown was placed in the lower third molar area of an 81-year-old female patient with complaints of dry and burning mouth. Salivary secretion was measured, and the patient was asked to fill in written satisfaction questionnaires. The patient was monitored for a year, comparing her salivary secretion rates and the written questionnaires. RESULTS: The results showed a constant slight but significant increase in the salivary secretion and in the patient's personal feelings as presented in the questionnaires. CONCLUSIONS: The saliva stimulation device Saliwell Crown, placed on an implant in an 81-year-old patient with dry and burning mouth complaints, presented promising results when both the salivary secretion tests and the self assessment questionnaires were examined and compared. PMID- 19681935 TI - Accelerated photo-induced hydrophilicity promotes osseointegration: an animal study. AB - BACKGROUND: In the previous in vitro study, fluoride-modified, anodized porous titanium was proven to have enhanced its photo-induced hydrophilicity, which induced the hyperactivation of initial cell response. PURPOSE: The purpose of the present study was to investigate in vivo bone apposition during the early stages of osseointegration in rabbit tibiae. MATERIALS AND METHODS: Anodized porous titanium implants (TiU, TiUnite(r), Nobel Biocare AB, Goteborg, Sweden) were modified with 0.175 wt% ammonium hydrogen fluoride solution (NH(4) F-HF(2) ). Twenty-four hours prior to the experiments, the surface-modified implants were ultraviolet-irradiated (modTiU). Blinded and unpackaged TiU implants were used as controls. Thereafter, the implants were placed in the rabbit tibial metaphyses and histomorphometrically analyzed at 2 and 6 weeks after insertion. RESULTS: ModTiU demonstrated a significantly greater degree of bone-to-metal contact than TiU after 2 and 6 weeks of healing. CONCLUSION: The results proved that the enhanced photo-induced hydrophilicity of the NH(4) F-HF(2) -modified anodized implants promoted bone apposition during the early stages of osseointegration. PMID- 19681936 TI - The "dome phenomenon" associated with maxillary sinus augmentation. AB - AIM: This case series is aimed to report a new phenomenon, the "dome phenomenon," which was observed in infected augmented sinuses over several years. METHODS: Five patients are presented in whom sinus lift augmentation resulted in postoperative infection with inflammation and suppuration. The patients received aggressive anti-inflammatory treatment and surgical debridement of the inflamed tissue, including some grafted material performed through the lateral window of the primary procedure. RESULTS: The inflammatory condition was reversed, and the site healed clinically. Radiographically, a dome-shaped, radio-opaque tissue was observed at the superior most aspect of the grafted sinus. This "dome phenomenon" was further confirmed during dental implant placement, which indicates healing potential adjacent to the maxillary sinus membrane. CONCLUSIONS: The current report, as well as other studies and case series, suggests that there is great potential for healing and bone formation in the maxillary sinus membrane. The precise mechanism is not known. Further clinical and histologic studies are needed to understand the regenerative potential of the Schneiderian membrane. PMID- 19681937 TI - Minimally invasive antral membrane balloon elevation - results of a multicenter registry. AB - BACKGROUND AND PURPOSE: Frequently, the posterior maxilla lacks sufficient bone mass to support dental implants. This multiphysician registry assessed the feasibility and safety of minimally invasive antral membrane balloon elevation (MIAMBE), followed by bone augmentation and implant fixation. MATERIALS AND METHODS: One hundred twelve consecutive patients were referred for MIAMBE. Following pre-procedural assessment and informed consent, patients underwent alveolar crest exposure, and 3 mm osteotomy followed by MIAMBE. Platelet-rich fibrin and bone substitutes were injected under the antral membrane; implant placement and primary closure were executed at the same sitting. Implant loading was carried out 6 to 9 months later. RESULTS: One hundred nine (97.3%) patients successfully concluded the initial procedure. Three patients had membrane tear requiring procedure abortion. One case of infection was documented at 4 weeks. Procedure time was 58 +/- 23 minutes. Incremental bone height consistently exceeded 10 mm, and implant survival of 95% was observed at 6 to 9 months. CONCLUSION: MIAMBE can be applied to all patients in need of posterior maxilla bone augmentation with high procedural success, low complication rate, and satisfactory bone augmentation and implant survival. As it is minimally invasive and associated with minimal discomfort, MIAMBE should be an alternative to the currently employed methods of maxillary bone augmentation. PMID- 19681938 TI - Inlay versus onlay iliac bone grafting in atrophic posterior mandible: a prospective controlled clinical trial for the comparison of two techniques. AB - PURPOSE: To compare the efficacy of inlay and onlay bone grafting techniques in terms of vertical bone formation and implant outcomes for correcting atrophic posterior mandibles. MATERIALS AND METHODS: Twenty surgical sites were assigned to two treatment groups, inlay and onlay, with iliac crest as donor site. After 3 to 4 months, 43 implants were placed and loaded 4 months later. The median follow up after loading was 18 months. RESULTS: For the inlay versus onlay group, median bone gain was 4.9 versus 6.5 mm (p = .019), median bone resorption was 0.5 versus 2.75 mm (p < .001), and median final vertical augmentation was 4.1 versus 4 mm (p = .190). The implant survival rate was 100% in both groups, while the implant success rate was 90% versus 86.9% (p = .190, not significant). A minor and major complication rate of 20% and 10%, respectively, for both groups was encountered. CONCLUSIONS: Inlay results in less bone resorption and more predictable outcomes, but requires an experienced surgeon. In contrast, onlay results in greater bone resorption and requires a bone block graft oversized in height, but involves a shorter learning curve. Once implant placement has been carried out, the outcomes are similar for both procedures. PMID- 19681939 TI - Evaluation of effects of topic melatonin on implant surface at 5 and 8 weeks in Beagle dogs. AB - PURPOSE: The aim of this study was to evaluate the effect of the topical application of melatonin on osteointegration of dental implants in Beagle dogs 5 and 8 weeks after their insertion. MATERIALS AND METHODS: For subsequent insertion of dental implants, upper and lower premolars and molars were extracted from 12 Beagle dogs. Each mandible received cylindrical screw implants of 3.25 mm in diameter and 10 mm in length. The implants were randomly assigned to the mesial and distal sites on each side of the mandible. Prior to implanting, 1.2 mg lyophylized powder melatonin was applied to one bone hole at each side of the mandible. None was applied at the control sites. Eight histological sections per implant were obtained for histomorphometric studies. RESULTS: After 5- and 8-week treatment periods, melatonin significantly increased the inter-thread bone (p < 0.05) and new bone formation (p < 0.05) in comparison to control implants in both weeks. There were no significant increases in the bone-to-implant contact and peri-implant bone (p > 0.05). CONCLUSION: Topical application of melatonin may act as a biomimetic agent in the placement of endo-osseous dental implants at 5 and 8 weeks after the implantation. PMID- 19681940 TI - Immediate loading of two implants with a mandibular implant-retained overdenture: a new treatment protocol. AB - PURPOSE: The aim of this study was to present the clinical outcomes of the immediate loading of two bar-splinted implants retaining a mandibular overdenture. MATERIALS AND METHODS: In a clinical trial, 124 edentulous patients were treated according to a new treatment concept, which involves the immediate loading of two bar-splinted SLActive implants with an implant-retained mandibular overdenture. The new conventional mandibular denture is used as a template for implant positioning and as an impression tray, and for mounting the retention clip by the dental laboratory. At the same day the implants are placed, the conventional denture is converted into an implant-retained overdenture. During the healing and evaluation period, resonance frequency analysis (RFA) was undertaken to assess the effect of loading on implant stability and survival. RESULTS: The survival rate of the implants was 98.8% during the evaluation period (12-40 months). Only 3 of the 248 implants were lost. During the healing (osseointegration) phase, the implant-stability quotient increased significantly (p = .0001). During the evaluation period, four patients (3%) needed a relining of their mandibular overdenture, whereas 13 patients (11%) needed relining of the maxillary denture. CONCLUSIONS: Two interconnected implants can be successfully loaded by a mandibular overdenture at the same day of implant placement with a high survival rate of the implants. Only a few patients needed additional relining of the overdenture. Repeated RFA measurements can be useful in gauging implant stability and survival. PMID- 19681941 TI - Infrared thermographic evaluation of temperature modifications induced during implant site preparation with cylindrical versus conical drills. AB - BACKGROUND: A few studies have investigated the influence of drilling on bone healing. Many factors have been reported to influence temperature rise during surgical preparation for implant placement: drill geometry, drilling depth, sharpness of the cutting tool, drilling speed, pressure applied to the drill, use of graduated versus one-step drilling, intermittent versus continuous drilling, and use or not of irrigation. PURPOSE: The objective of this study was to quantify the temperature changes in cortical bone and at the apical portion of the drills during implant site preparation with a cylindrical implant drill versus a conical implant drill. MATERIALS AND METHODS: Two implant drill systems were evaluated in vitro using bovine femoral cortical bone. The two implant drill systems evaluated in this study were system A (a cylindrical drill with triple twist drills) (Bone System, Milano, Italy) and system B (a conical drill with quadruple twist drills) (Bone System). Site preparation began, and the temperature of the cortical bone and at the apical portion of the drill was measured by the infrared thermography. RESULTS: The mean temperature produced in the cortical bone during implant preparation was 31.2 +/- 0.5 degrees C for the cylindrical drills and 29.1 +/- 0.6 degrees C for the conical drill. The mean temperature produced in the apical portion of the drill during implant site preparation was 32.1 +/- 0.7 degrees C for the cylindrical drill system and 29.6 +/- 0.6 degrees C for the conical drill. Statistically significant differences were found in the temperature measurements in the cortical bone in the two groups (p < .05). A statistically significant difference was observed for the temperature measurements in the apical portion of the drill in the two groups (p < .005). DISCUSSION: The model system used in this work was able to evaluate the temperature in the cortical bone and in the apical portion of the drills; the temperature modifications in the apical portion of the drill seemed to be correlated to the drill geometry. The results of the present study showed that drill geometry seems to be an important factor in heat generation during implant site preparation. CONCLUSION: The drill geometry could explain the increased temperature in the apical portion of the drill. PMID- 19681942 TI - Histological outcomes on the development of new space-making devices for maxillary sinus floor augmentation. AB - BACKGROUND: Previous studies have pointed out that the mere elevation of the maxillary sinus membrane promotes bone formation without the use of augmentation materials. PURPOSE: This experimental study aimed at evaluating if the two-stage procedure for sinus floor augmentation could benefit from the use of a space making device in order to increase the bone volume to enable later implant installation with good primary stability. MATERIALS AND METHODS: Six male tufted capuchin primates (Cebus apella) were subjected to extraction of the three premolars and the first molar on both sides of the maxilla to create an edentulous area. The sinuses were opened using the lateral bone-wall window technique, and the membrane was elevated. One resorbable space-making device was inserted in each maxillary sinus, and the bone window was returned in place. The animals were euthanatized after 6 months, and biopsy blocks containing the whole maxillary sinus and surrounding soft tissues were prepared for ground sections. RESULTS: The histological examination of the specimens showed bone formation in contact with both the schneiderian membrane and the device in most cases even when the device was displaced. The process of bone formation indicates that this technique is potentially useful for two-stage sinus floor augmentation. The lack of stabilization of the device within the sinus demands further improvement of space-makers for predictable bone augmentation. CONCLUSIONS: It is concluded that (1) the device used in this study did not trigger any important inflammatory reaction; (2) when the sinus membrane was elevated, bone formation was a constant finding; and (3) an ideal space-making device should be stable and elevate the membrane to ensure a maintained connection between the membrane and the secluded space. PMID- 19681944 TI - Relationship between systemic bone mineral density and local bone quality as effectors of dental implant survival. AB - PURPOSE: This study aimed to assess (1) the relationship of systemic bone mineral density (BMD) and osteoporotic status with the surgeon's subjective assessment of local jawbone quality, and (2) whether the surgeon's subjective assessment of local jawbone quality is a predictor of implant failure. MATERIALS AND METHODS: A retrospective analysis of 2,867 dental implants placed in 645 patients was accomplished. The surgeon's assessment of bone quality at the time of dental implant placement was recorded. Of those, 208 patients with 701 implants had BMD data available within 3 years. Statistical analyses were conducted to determine relationships between BMD, osteoporotic status, and local jawbone quality and to determine the relationship between local jawbone quality and implant survival. RESULTS: There was no association between systemic BMD and the surgeon's assessment of bone quality (p =.52) nor between osteoporotic status and the surgeon's assessment of local jawbone quality (Spearman rank correlation coefficient=0.08). Additional retrospective analysis revealed implants placed in moderate- (hazard ratio=1.67; p=.043) or poor-quality (HR=3.45, p< .001) bone (surgeon's assessment) were significantly more likely to fail than implants placed in good-quality bone. CONCLUSION: Systemic BMD and osteoporotic status are not associated with local jawbone quality. Implants placed in good-quality bone, as assessed subjectively by the surgeon at the time of implant placement, have significantly better survival characteristics than implants placed in moderate /poor-quality bone. PMID- 19681943 TI - Nanoporous TiO(2) thin film on titanium oral implants for enhanced human soft tissue adhesion: a light and electron microscopy study. AB - BACKGROUND: Previous experimental studies have demonstrated direct soft tissue attachment for nanoporous titanium dioxide (TiO(2) ) thin film on implants, while implants without TiO(2) thin film have not shown this capability. PURPOSE: The aims were to evaluate and compare TiO(2) surface-modified experimental microimplants with unmodified microimplants with respect to tissue interaction of the human oral mucosa evaluated by light microscopy on ground sections and semithin sections and transmission electron microscopy on ultrathin sections, and to characterize the inflammatory response and the level of the marginal bone resorption. MATERIALS AND METHODS: The study was a single-center, randomized, comparative, clinical investigation with intrasubject comparison of implants with and without TiO(2) thin film in 15 patients. RESULTS: Two comparator microimplants showed mild erythema and expulsion of fluids. The surrounding tissues around all test implants were clinically healthy. The oral mucosa in contact with the abutment part of the microimplant was 72% for the test implants and 48% for the comparator implants, a statistically significant difference (p =.0268). No statistically significant difference was found in other histological variables. The marginal bone loss in 14 weeks was 0.5 mm for the stable test (n = 11) and 1.7 mm for the stable comparator implants (n = 9; p = .0248). CONCLUSIONS: The nanoporous TiO(2) surface modification has potential clinical benefits because of increased adherence of soft tissue and possible reduced bone resorption. PMID- 19681945 TI - Fosamprenavir treatment in a highly active antiretroviral therapy schedule induces a HCV-RNA decrease and a Th1 network boost in HIV/HCV-coinfected patients. AB - HIV/HCV co-infected naive patients (four females and six males) were evaluated for their response to the following treatment schedule: [(AZT 300 mg + 3TC 300 mg twice daily) + (fosamprenavir 700 mg twice daily) + (RTV 100 mg)]. CD3+/CD4+ T cells, interferon-gamma (INF-gamma) and interleukin-4 (IL-4) HCV-specific response, viral loads and transaminase levels were evaluated at time 0, and after 1, 3 and 6 months of therapy (T0, T1, T3, and T6 respectively). HIV-RNA, HCV-RNA and transaminases decreased at T1 and T3 compared with T0 (Mann-Whitney p <0.001, p <0.01 and p <0.01, respectively). At all time points, CD4+ and HCV-specific INF gamma responses were higher (p <0.001; p <0.001), and IL-4 lower (p <0.01) after treatment. At T6, HCV-RNA was only negative in four out of ten patients whereas all had normal transaminase levels. These findings indicate that HAART treatment including fosamprenavir is able to activate a Th1 network in HIV/HCV co-infected patients. Moreover, these results, to be confirmed by larger cohort follow-up studies, suggest that this protease inhibitor could have potential implications for the treatment of chronic hepatitis C in HIV-positive patients. PMID- 19681946 TI - Adult invasive pneumococcal disease between 2003 and 2006 in North-Rhine Westphalia, Germany: serotype distribution before recommendation for general pneumococcal conjugate vaccination for children <2 years of age. AB - A laboratory-based surveillance study of adult invasive pneumococcal disease was conducted in North-Rhine Westphalia, Germany's most populous federal state, with approximately 18 million inhabitants. Invasive isolates (n = 519) were obtained between 2003 and 2006, before the general recommendation for vaccination of German children <2 years with the pneumococcal conjugate vaccine was issued at the end of July 2006. Penicillin G resistance was observed in 5% of meningitis cases. In the non-meningitis group, only intermediately resistant strains were detected (0.4%). Intermediate resistance to cefotaxime occurred both in meningitis cases (1.7%) and non-meningitis cases (0.4%). Non-susceptibility rates (intermediate resistance and resistance) were 16.2% for macrolides, 10.9% for trimethoprim-sulphamethoxazole, 5.0% for tetracycline, 3.9% for clindamycin, and 0.4% for levofloxacin. All isolates were susceptible to amoxycillin (non meningitis) and telithromycin. The leading serotypes were serotypes 14 (14.3%), 7F (9.4%), 3 (9.2%), 4 (8.7%) and 1 (8.1%). Serotype coverage for the seven valent conjugate vaccine was 43.9%. For the ten-valent and 13-valent vaccines (in development), the coverages were 61.8% and 76.7%, respectively. The 23-valent polysaccharide vaccine had a coverage of 91.1%. PMID- 19681947 TI - Sputum colour reported by patients is not a reliable marker of the presence of bacteria in acute exacerbations of chronic obstructive pulmonary disease. AB - Sputum colour is regarded as a good marker of bacterial involvement in acute exacerbations of chronic obstructive pulmonary disease (COPD) and guides many physicians in deciding on antibiotic treatment. Although most doctors rely on the sputum colour that is reported by patients, it can also be assessed using a validated colour chart. In this study, reported sputum colour and assessed sputum colour were compared as markers of the presence of bacteria, bacterial load, and systemic inflammation. Data on 257 exacerbations in 216 patients hospitalized with an acute exacerbation were analysed (mean age, 72 years; mean forced expiratory volume in 1 s, 44.8% + or - 17.8% (+ or - standard deviation)). Sputum colour was reported by the patients and assessed at the laboratory with a colour chart. Subsequently, quantitative sputum cultures were performed. C-reactive protein was measured as a marker of systemic inflammation. A sputum sample was obtained in 216 exacerbations (84%), of which 177 (82%) were representative. A pathogen was identified in 155 patients (60%). Assessed sputum colour was a better marker of the presence of bacteria (OR 9.8; 95% CI 4.7-20.4; p <0.001) than reported sputum colour (OR 1.7; 95% CI 1.0-3.0; p 0.041). The sensitivity and specificity were 73% and 39% for reported sputum colour, and 90% and 52% for assessed sputum colour. Assessed sputum colour was clearly related to sputum bacterial load and C-reactive protein levels, whereas reported sputum colour was not. It is concluded that sputum colour reported by patients is an unreliable marker of the presence of bacteria in acute exacerbations of COPD. Assessed sputum colour is clearly superior and is also related to bacterial load and systemic inflammation. PMID- 19681948 TI - Panton-Valentine Leucocidin-related disease in England and Wales. AB - Within the framework of the Health Protection Agency's programme of enhanced surveillance of Staphylococcus aureus with Panton-Valentine Leucocidin (PVL-SA) in England and Wales conducted during 2005-2006, we identified 720 PVL-SA, representing a two-fold increase between 2005 (n = 224) and 2006 (n = 496). The number of PVL-methicillin-resistant S. aureus rose from 119 to 159 in that period. Isolates were referred by 112 centres and included outbreaks of PVL related disease in community and healthcare settings. One hundred individuals had systemic disease symptoms. Planned systematic surveillance-based studies aim to better address the question of whether these increases reflect an increasing prevalence of PVL-SA and/or improved case ascertainment of PVL-related syndromes. PMID- 19681950 TI - Determinants of Moraxella catarrhalis colonization in healthy Dutch children during the first 14 months of life. AB - Moraxella catarrhalis is an established bacterial pathogen, previously thought to be an innocent commensal of the respiratory tract of children and adults. The objective of this study was to identify significant risk factors associated with M. catarrhalis colonization in the first year of life in healthy Dutch children. This study investigated a target cohort group of 1079 children forming part of the Generation R Study, a population-based prospective cohort study following children from fetal life until young adulthood, conducted in Rotterdam, The Netherlands. Nasopharyngeal swabs for M. catarrhalis culture were obtained at 1.5, 6 and 14 months of age, with all three swabs being available for analyses from 443 children. Data on risk factors possibly associated with M. catarrhalis colonization were obtained by questionnaire at 2, 6 and 12 months. M. catarrhalis colonization increased from 11.8% at age 1.5 months to 29.9% and 29.7% at 6 and 14 months, respectively. Two significantly important colonization risk factors were found: the presence of siblings and day-care attendance, which both increased the risk of being positive for M. catarrhalis colonization on two or more occasions within the first year of life. Colonization with M. catarrhalis was not associated with gender, educational level of the mother, maternal smoking, breast-feeding, or antibiotic use. Apparently, crowding is an important risk factor for early and frequent colonization with M. catarrhalis in the first year of life. PMID- 19681949 TI - Higher rates of streptococcal colonization among children in the Pacific Rim Region correlates with higher rates of group A streptococcal disease and sequelae. AB - Group A streptococcal (GAS) pharyngeal colonization rates were determined among 1061 asymptomatic students in Hawaii and American Samoa where acute rheumatic fever rates are high. All GAS isolates were emm sequence typed. Although pharyngeal colonization rates were low in Hawaii (3.4%), Pacific Islander children had significantly higher colonization rates (5.7% vs. 1.2% in other ethnic groups, p <0.05). The colonization rate was higher in American Samoa (13%). Few emm types that were infrequently observed in symptomatic infections in Hawaii were repeatedly identified in both sites. These emm types were previously described among asymptomatic children suggesting a type-specific association with pharyngeal colonization. PMID- 19681951 TI - Kinetics of nasopharyngeal shedding of novel H1N1 (swine-like) influenza A virus in an immunocompetent adult under oseltamivir therapy. AB - We describe a patient with confirmed novel H1N1 (swine-like) influenza A virus who had daily nasal swabs tested during oseltamivir therapy. Nasal shedding remained positive for 2 days and became negative on day 3. This report presents the first available data on the kinetics of shedding of this novel virus under antiviral therapy. PMID- 19681952 TI - Rapid identification and antimicrobial susceptibility testing of Gram-positive cocci in blood cultures by direct inoculation into the BD Phoenix system. AB - Rapid identification and antimicrobial susceptibility testing (AST) of the causative agent(s) of bloodstream infections are essential for the selection of appropriate antimicrobial therapy. To speed up the identification and AST of the causative agent, the fluid from blood culture bottles of a Bactec 9240 instrument (Becton Dickinson) containing Gram-positive cocci was mixed with saponin. After a 15-min incubation, the bacteria were harvested and transferred to the appropriate panel of a BD Phoenix automated microbiology system (Becton Dickinson) for identification and AST. With this approach (referred to as the direct method), we concordantly/correctly identified 56 (82%) of 68 monomicrobial cultures using the results obtained with the method currently used in our laboratory (current method) as comparator. Two (3%) isolates could not be identified and ten (15%) were misidentified. Complete agreement, concerning clinical susceptibility categories and MIC values, between the AST results determined with the direct method and the current method was found for 32 (55%) of 58 isolates. The E-test indicated that the direct method yielded a correct susceptibility profile for 13 of the remaining 26 blood culture isolates. Therefore, a concordant/correct susceptibility profile (with all antimicrobial agents tested) was obtained for 45 (77%) of 58 cultures. The overall error rate amounted to 1.9%, with the majority (1.3%) of errors being minor. Importantly, the results obtained with the direct method were available 12-24h earlier than those obtained with the current method. PMID- 19681954 TI - Fatal Clostridium difficile enteritis caused by the BI/NAP1/027 strain: a case series of ileal C. difficile infections. AB - Clostridium difficile generally causes diarrhoea and colitis. Small-bowel infections are considered to be rare. Twelve cases of ileal C. difficile infections are presented, including the first reported case proven to be caused by the hypervirulent BI/NAP1/027 strain. This case series suggests that small bowel involvement in C. difficile infections may be more frequent than previously thought. PMID- 19681953 TI - Detection of residual human immunodeficiency virus type 1 reverse transcriptase K103N minority species in plasma RNA and peripheral blood mononuclear cell DNA following discontinuation of non-nucleoside therapy. AB - Non-nucleoside reverse transcriptase inhibitor (NNRTI) therapy failed in 30 patients with the typical human immunodeficiency virus type 1 reverse transcriptase K103N mutation, detected using standard genotyping. Following discontinuation of NNRTI therapy for a median of 55.9 weeks and a decrease of K103N mutant species to undetectable levels in plasma RNA, minority K103N species remained detectable, by allele-specific PCR, for longer periods of time and at higher frequency, in peripheral blood mononuclear cell (PBMC) DNA than in plasma RNA (76.7% and 46.7% of samples with residual K103N species detected at median frequencies of 18.0% and 3.8%, respectively). Analysis of PBMC DNA should be considered when searching for residual K103N mutant species in patients previously exposed to NNRTIs. PMID- 19681955 TI - Genetic diversity in CC398 methicillin-resistant Staphylococcus aureus isolates of different geographical origin. AB - Staphylococcus aureus of sequence type 398 has emerged in Europe, North America and Asia, and has typically been associated with livestock and their human contacts. We analysed two Panton-Valentine leukocidin (PVL)-negative t034-ST398 isolates from humans in contact with pigs and two t034-ST398 PVL-positive isolates from two unrelated, adopted Chinese children, using multistrain microarrays to determine genomic variability between the two sets of isolates. The ST398 isolates clearly belong to the same lineage when compared to other clonal lineages. However, the four isolates cluster into two distinct groups corresponding to differences in epidemiology based on mobile genetic elements and resistance patterns, suggesting that the two groups are epidemiologically distinct. PMID- 19681956 TI - Association of the bla(CMY-10) gene with a novel complex class 1 integron carrying an ISCR1 element in clinical isolates from Korea. AB - The bla(CMY-10) gene responsible for beta-lactam resistance was located on a new complex class 1 integron within a conjugative plasmid. The sul1-type class 1 integron, containing an aadA2a gene cassette, was identified upstream of bla(CMY 10). A unique gene array (yqgF-yqgE-gshB-orf97--orf105) was identified downstream of bla(CMY-10.). PMID- 19681957 TI - Extended-spectrum and CMY-type beta-lactamase-producing Escherichia coli in clinical samples and retail meat from Pittsburgh, USA and Seville, Spain. AB - Infections due to Escherichia coli producing extended-spectrum beta-lactamase (ESBL) or CMY-type beta-lactamase (CMY) are increasingly observed in non hospitalized patients. The origin of these organisms is uncertain, but retail meat contaminated with E. coli may be a source. In the present study, clinical information and strains collected from patients infected or colonized with ESBL producing and CMY-producing E. coli at hospitals in Pittsburgh, USA and Seville, Spain were investigated. Retail meat purchased in these cities was also studied for the presence of these organisms. Twenty-five and 79 clinical cases with ESBL producing E. coli and 22 cases and one case with CMY-producing E. coli were identified in Pittsburgh and Seville, respectively. Among them all, community acquired and healthcare-associated cases together constituted 60% of the cases in Pittsburgh and 73% in Seville. Community-acquired cases were more common in Seville than in Pittsburgh (49% vs. 13%; p <0.001). ESBL-producing and CMY producing E. coli isolates were commonly recovered from the local retail meat. In particular, 67% (8/12) of retail chickens in Seville and 85% (17/20) of those in Pittsburgh contained ESBL-producing and CMY-producing E. coli isolates, respectively. Among the ESBL-producing isolates, CTX-M and SHV were the most common ESBL types in both clinical and meat isolates. Approximately half of the ESBL-producing and CMY-producing E. coli isolates from meat belonged to phylogenetic groups associated with virulent extra-intestinal infections in humans. Community and healthcare environments are now significant reservoirs of ESBL-producing and CMY-producing E. coli. Retail meat is a potential source of these organisms. PMID- 19681958 TI - Transthoracic fine-needle aspiration in the aetiological diagnosis of community acquired pneumonia. AB - To investigate the safety and practicability of conducting transthoracic fine needle aspiration (TFNA) in a general hospital setting, we applied the TFNA procedure to 20 patients hospitalized with community-acquired pneumonia (CAP) within 36 h of admission. Also, a preliminary assessment was made of the potential value of adding TFNA to conventional methods of diagnostic microbiology. TFNA was easy to perform and caused little discomfort, and no serious adverse events were observed. In spite of ongoing antimicrobial treatment, a likely aetiological diagnosis was established for 14 of 20 (70%) of the patients. TFNA may provide important additional information on the aetiology of CAP. PMID- 19681959 TI - Molecular characterization and susceptibility of methicillin-resistant and methicillin-susceptible Staphylococcus aureus isolates from hospitals and the community in Vladivostok, Russia. AB - A prospective study was conducted during an 8-month period, from August 2006 to April 2007, to describe the epidemiology of Staphylococcus aureus-associated infections. In addition, the molecular characteristics, antimicrobial susceptibilities and antibiotic resistance determinants were identified in S. aureus isolates from hospitals and the community in Vladivostok, Russia. Among the 63 S. aureus isolates eligible for this study, methicillin resistance was observed in 48% (n = 30). Hospital-acquired strains accounted for 93% (28/30) of all methicillin-resistant S. aureus (MRSA) isolates. The major MRSA clone (sequence type (ST) 239, staphylococcal cassette chromosome mec (SCCmec) type III, Panton-Valentine leukocidin (PVL)-negative, with two related staphylococcal protein A gene (spa) types (types 3 and 351)) represented 90% of all of the MRSA isolates. This clone was multidrug-resistant, and 41% of isolates showed resistance to rifampicin. Community-acquired MRSA isolates (n = 2) were categorized as ST30, SCCmecIV, spa type 19, and PVL-positive, and as ST8, SCCmecIV, of a novel spa type 826, and PVL-negative. Eight different STs were detected among methicillin-susceptible S. aureus (MSSA) isolates, of which 55% were PVL-positive. One MSSA clone, which was categorized as ST121, spa type 273, and PVL-positive, caused fatal community-acquired pneumonia infections. The strains predominantly isolated in hospitals in Russia belonged to the multidrug resistant Brazilian/Hungarian ST239 MRSA clone; however, this clone has new antibiotic susceptibilities. Additionally, the emergence of PVL-positive MSSA strains with enhanced virulence was observed, warranting continued surveillance. PMID- 19681960 TI - Prevalence and clinical aspects of human bocavirus infection in children. AB - Human bocavirus (HBoV) was recently described as a new member of the Parvoviridae. In order to investigate the suggested association of HBoV with respiratory and gastric disease in infants and young children, sera of 357 paediatric patients hospitalized with infectious and non-infectious diseases were retrospectively analyzed for the presence of HBoV DNA and virus-specific antibodies using quantitative PCR and ELISA, respectively. HBoV seroprevalence was determined to range from 25% in infants younger than 1 year of age to 93% in children aged more than 3 years. Viral loads between 1 x 10(2) and 1.2 x 10(6) geq/mL were observed in 6.7% (20/297) of sera obtained preferentially from young children suffering from infectious diseases. HBoV genomes were furthermore detected in 5% (3/60) of sera collected from individuals with non-infectious illnesses. HBoV DNA was present most frequently in patients with respiratory disease (9.6%). Whereas only 5.2% of patients with upper respiratory tract disease were viraemic, HBoV DNA was found in 14.6% and 10.0% of patients with lower respiratory tract illness and pneumonia, respectively. Acute HBoV infections were also observed in 7.5% of patients with gastroenteritis and in one child with inflammatory bowel disease. None of 77 patients hospitalized for various other infectious diseases (e.g. rash, urinary tract infection, meningitis) displayed viraemia. In 60.9% and 47.8% of DNA-positive children, HBoV specific IgM and IgG was observed, respectively. The present prospective study provides comprehensive data on the clinical association of acute HBoV infection with respiratory illness and on the seroprevalence of virus-specific antibodies in children. PMID- 19681962 TI - Accuracy of a newly developed integrated system for dental implant planning. AB - OBJECTIVES: To evaluate the accuracy of the first integrated system for cone-beam CT (CBCT) imaging, dental implant planning and surgical template-aided implant placement. MATERIALS AND METHODS: On the basis of CBCT scans, a total of 54 implant positions were planned for 10 partially edentulous anatomical patient equivalent models. Surgical guides were ordered from the manufacturer (SICAT). Two different types of guidance were assessed: for assessment of the SICAT system inherent accuracy vendor's titanium sleeves of 2 mm internal diameter and 5 mm length were utilized for pilot drills. The guide sleeves of the NobelGuide system were implemented for fully guided surgery and implant insertion. Deviations perpendicular to the implant axes at the crestal and apical end, as well as the angle deviations between the virtual planning data and the surgical results, were measured utilizing a follow-up CBCT investigation and referential marker-based registration. RESULTS: The SICAT system inherent mean deviation rates for the drilled pilot osteotomies were determined to be smaller than 500 mum even at the apical end. Mean angle deviations of 1.18 degrees were determined. Utilizing the NobelGuide sleeve-in-sleeve system for fully guided implant insertion in combination with the investigated template technology enabled to insert dental implants with the same accuracy. Crestal deviations, in general, were significantly lower than the apical deviations. CONCLUSION: Although hardly comparable due to different study designs and measurement strategies, the investigated SICAT system's inherent accuracy corresponds to the most favourable results for computer-aided surgery systems published so far. In combination with the NobelGuide surgical set for fully guided insertion, the same accuracy level could be maintained for implant positioning. PMID- 19681961 TI - Tuberculosis transmission patterns among Spanish-born and foreign-born populations in the city of Barcelona. AB - During a 2-year period (2003-2004), tuberculosis (TB) transmission in Barcelona and the factors related to transmission among the Spanish- and foreign-born populations were studied by molecular epidemiology. Data were obtained from TB cases and Conventional Contact Tracing registries and genotyping was performed using restriction fragment length polymorphism (RFLP)-IS6110 and MIRU12 as a secondary typing method. Of the 892 TB cases reported, 583 (65.3%) corresponded to Spanish-born and 309 (34.6%) to foreign-born. Six hundred and eighty-seven cases (77%) were confirmed by culture. RFLP typing of 463/687 (67.4%) isolates was performed, revealing 280 (60.5%) unique and 183 (39.5%) shared patterns, which were grouped into 65 clusters. Spanish-born individuals were significantly more clustered than foreign-born individuals (44.6% vs. 28.8%; p 0.016). Clustering in foreign-born individuals was associated with HIV (p 0.051, odds ratio = 3.1, 95% confidence interval 1-10.9) and alcohol abuse (p 0.022), whereas, in the Spanish-born individuals, clustering was associated with age in the range 21-50 years, (p 0.024). Of the total clusters, 36/65 (55.3%) included only Spanish-born patients, whereas 22/65 (33.8%) included individuals from both populations. In mixed clusters, the index case was Spanish-born in 53% and foreign-born in 47%. Among the foreign-born, 2.8% were ill on arrival, 30% developed TB within the first year and 50.3% developed TB within the first 2 years; 58.3% were from South America. In conclusion, half of the foreign-born TB patients developed the disease during the first 2 years after arrival, which, in most cases, was the result of endogenous reactivation. Recent TB transmission among Spanish-born and foreign-born populations, as well as bidirectional transmission between communities, contributed significantly to the burden of TB in Barcelona, suggesting the need to improve Public Health interventions in both populations. PMID- 19681963 TI - Peri-implant parameters in head and neck reconstruction: influence of extraoral skin or intraoral mucosa. AB - OBJECTIVE: This study is designed to assess dental implants supporting overdentures in edentulous patients with operated head and neck malignancies using parameters to detect peri-implant disease. MATERIAL AND METHODS: Thirty four implants supporting overdentures in 34 oral cancer patients were examined. Clinical parameters [plaque index, probing depth, bleeding on probing (BOP), origin of peri-implant soft tissue, and amount of irradiation] were recorded, and microbiological identification of periodontal pathogens was carried out by DNA DNA hybridization. To identify yeast species, the samples were cultivated on Sabouraud agar plates and subsequently identified by API 20C AUX plates. An implant site showing BOP, probing pocket depth (PPD)>or=5 mm and radiographic vertical bone loss was considered to have peri-implant disease. RESULTS: Colonization by periodontal pathogens was found on 15 implants, while yeast species were found in 14 cases. Using a univariate analysis, none of the investigated parameters (microbiologic sign, detection of yeast, origin of peri implant soft tissue and irradiation) were significantly correlated to signs of peri-implant disease. In the multivariate analysis, yeast [odds ratio (OR) 12.32, P=0.033] and periodontal pathogen (OR 9.88, P=0.046) were significant predictor variables for peri-implant disease. Yeasts were less frequently detected around implants placed in re-vascularized skin flaps if irradiation was set as a confounder (P=0.019). CONCLUSIONS: With respect to the pilot study nature of the study peri-implant soft tissue origin and irradiation had little influence on the development of peri-implant disease. Yeast and periodontal pathogen were explanatory variables for the development of peri-implant disease. Considering the effect of irradiation on the prevalence of yeast, yeast was less frequently observed in peri-implant soft tissue of the skin. Based on these data, future studies on the role of yeast and soft tissue in peri-implant disease should be encouraged. PMID- 19681964 TI - Implant stability and bone density: assessment of correlation in fresh cadavers using conventional and osteotome implant sockets. AB - OBJECTIVE: To compare the primary stability of implants placed in conventional and osteotome sites and to evaluate the level of correlation between cutting torque measurements, resonance frequency analysis (RFA), and bone density. MATERIALS AND METHODS: Eight human femoral heads were scanned with computed tomography for bone density measurements as Hounsfield units (HU), and individualized computed tomography-based surgical stents were prepared for placement of implants. Five implant sockets were prepared in each collum (CoF), caput (CaF), and trochanter (Tr-MM) section of the femoral heads using the conventional drilling technique or by a combination of drilling and use of an osteotome. Cutting-torque values (CTV) of the implants were measured by a manual torque wrench, followed by determination of implant stability quotients (ISQ) by RFA. RESULTS: The CTVs of implants were similar in the conventional group, but different in the osteotome group (P<0.05). There was a general tendency toward achieving higher CTV and ISQ values in CoF than CaF and Tr-MM (P<0.05), and measurements in CaF and Tr-MM were comparable (P>0.05). The mean HU of sites were similar, although CoF had higher HU values (P>0.05). CTV of implants in CaF and Tr-MM and ISQ values in CoF in the conventional groups were higher than those in the osteotome groups (P<0.05). The correlation between CTV and HU in Tr-MM was significant in the osteotome group, although no other correlations between CTV, ISQ, and HU could be detected (P>0.025). CONCLUSIONS: Conventional placement led to higher implant stability than the drilling and osteotome technique used in the study. No correlation could be found between CTV, RFA, and bone density. PMID- 19681966 TI - Vertical ridge augmentation of the atrophic posterior mandible with interpositional bloc grafts: bone from the iliac crest vs. bovine anorganic bone. Clinical and histological results up to one year after loading from a randomized controlled clinical trial. AB - OBJECTIVES: To compare two different techniques for vertical bone augmentation of the posterior mandible: bone blocs from the iliac crest vs. anorganic bovine bone blocs used as inlays. MATERIALS AND METHODS: Ten partially edentulous patients having 5-7 mm of residual crestal height above the mandibular canal had their posterior mandibles randomly allocated to both interventions. After 4 months implants were inserted, and after 4 months, provisional prostheses were placed. Definitive prostheses were delivered after 4 months. Histomorphometry of samples trephined at implant placement, prosthesis and implant failures, any complication after loading and peri-implant marginal bone-level changes were assessed by masked assessors. All patients were followed up to 1 year after loading. RESULTS: Four months after bone augmentation, there was statistically significant more residual graft (between 10% and 13%) in the Bio-Oss group. There were no statistically significant differences in failures and complications. Two implants could not be placed in one patient augmented with autogenous bone because the graft failed whereas one implant and its prosthesis of the Bio-Oss group failed after loading. After implant loading only one complication (peri-implantitis) occurred at one implant of the autogenous bone group. In 16 months (from implant placement to 1 year after loading), both groups lost statistically significant amounts of peri-implant marginal bone: 0.82 mm in the autogenous bone group and 0.59 mm in the Bio-Oss group; however, there were no statistically significant differences between the groups. CONCLUSIONS: Both procedures achieved good results, but the use of bovine blocs was less invasive and may be preferable than harvesting bone from the iliac crest. PMID- 19681965 TI - Paste-like inorganic bone matrix: preclinical testing of a prototype preparation in the porcine calvaria. AB - OBJECTIVE: To evaluate the osteoconductive properties and the volume stability of an injectable paste-like inorganic bone matrix (PBM) in porcine calvaria defects. MATERIAL AND METHODS: We created six circumferential defects in the calvaria of 12 adult iberico pigs. The defects were filled with either PBM, Bio-Oss((R)) of different particle size, carrier alone, or left empty. PBM was composed of Bio Oss((R)) with a particle size ranging from 250 to 500 mum and a hydrogel-carrier of carboxymethylcellulose and collagen. After 6 and 12 weeks of healing, the animals were sacrificed and undecalcified ground sections were prepared and subjected to histologic and histomorphometric analysis. To quantify the osteoconductive properties of PBM, bone volume per tissue volume (BV/TV) in the defect area was determined. To determine the volume stability, bone substitute volume per tissue volume (BSV/TV) was measured. RESULTS: After 6 weeks, PBM particles in the center of the defect were surrounded by fibrous connective tissue, which was later replaced by bone. BV/TV in the PBM group increased from 29.7+/-12.7% (minimum 12.2%, maximum 43.7%) after 6 weeks to 43.9+/-14.9% (minimum 27.8%, maximum 63.9%) after 12 weeks (Mann-Whitney test; P=0.6). According to the Friedman test, BV/TV in groups containing Bio-Oss((R)) of different particle sizes, the carrier and the empty defects was similar to the results obtained with PBM (6 weeks P=0.8; 12 weeks P=0.22). BSV/TV in the PBM group was stable over time, with 10.1+/-9% (minimum 3.3%, maximum 27.6%) and 16.5+/-12.9% (minimum 1%, maximum 32.7%), after 6 and 12 weeks, respectively (P=0.72). BSV/TV in the PBM group was comparable to the results obtained with the Bio-Oss((R)) particles of different sizes (Friedman test; 6 weeks P=0.0503; 12 weeks P=0.56). CONCLUSIONS: The results of this preclinical study showed that the PBM is osteoconductive and maintains the augmented volume, similar to commercial Bio-Oss((R)). These data suggest that the osteoconductive properties of Bio Oss((R)) are maintained at the smaller particle size and in the presence of the carrier. PMID- 19681967 TI - Interproximal tissue dimensions in relation to adjacent implants in the anterior maxilla: clinical observations and patient aesthetic evaluation. AB - OBJECTIVES: This clinical study aimed to assess (i) interproximal tissue dimensions between adjacent implants in the anterior maxilla, (ii) factors that may influence interimplant papilla dimensions, and (iii) patient aesthetic satisfaction. MATERIAL AND METHODS: Fifteen adults, who had two or more adjacent implants (total of 35) in the anterior maxilla, participated in the study. The study design involved data collection from treatment records, clinical and radiographic assessment, and a questionnaire evaluating aesthetic satisfaction. RESULTS: The median vertical dimension of interimplant papillae, i.e., distance from tip of the papilla to the bone crest, was 4.2 mm. Missing papilla height (PH) at interimplant sites was on average 1.8 mm. Median proximal biologic width at interimplant sites was 7 mm. The most coronal bone-to-implant contact at implant-implant sites was located on average 4.6 mm apical to the bone crest at comparable neighbouring implant-tooth sites. The tip of the papilla between adjacent implants was placed on average 2 mm more apically compared with implant tooth sites. The contact point between adjacent implant restorations extended more apically by 1 mm on average compared with implant-tooth sites. Median missing PH was 1 mm when an immediate provisionalization protocol had been followed, whereas in the case of a removable temporary it was 2 mm. Split group analysis showed that for missing PH0.05). All substrates were densely covered by osteoblasts. ALP and osteocalcin production was similar on the examined surfaces. Cell morphology analysis revealed flat spread osteoblasts with cellular extensions on all substrates. CONCLUSIONS: These results indicate that ATZ may be a viable substrate for the growth and differentiation of human osteoblasts. Surface modification of ATZ by airborne particle abrasion alone or in combination with acid etching seems not to interfere with the growth and differentiation of the osteoblasts. PMID- 19681969 TI - Comparison of two bone substitute biomaterials consisting of a mixture of fibrin sealant (Tisseel) and MBCP (TricOs) with an autograft in sinus lift surgery in sheep. AB - OBJECTIVES: The aim of this study was to assess the efficacy and safety of macroporous biphasic calcium phosphate (MBCP())/fibrin grafts (TricOs((R)))/(Tisseel((R))) for sinus lift augmentation in sheep. Autologous bone grafts were used as a positive control, and dental implants were placed to assess the efficiency of the composite. MATERIALS AND METHODS: A total of 12 adult sheep (24 maxillary sinuses) were randomized to receive sinus lift augmentation with MBCP()/fibrin grafts obtained by either simultaneous or sequential addition of thrombin and fibrinogen to MBCP(), or autologous bone grafts. Six months post-sinus lift surgery, dental implants were placed. At 6 months post-sinus lift and 3 months after dental implant placement, the characteristics of newly formed bone and dental implant stability were assessed. The methods used were radiography, scanning electron microscopy, light microscopy, micro-CT analysis, radio frequency analysis (RFA), and image analysis. RESULTS: There were no clinical adverse events in the post-operative period. New bone formation was similar for MBCP()/fibrin grafts and autografts at 21-20% and 20%, respectively, at 6 months, and at 34-35% and 35%, respectively, at 9 months. Implantability of dental implants was better at the time of placement with MBCP()/fibrin grafts than autografts at 81-88% and 69%, respectively. Three months after placement, RFA showed better implantability with MBCP()/fibrin grafts than with autografts at 75-82% and 71%, respectively. The bone contact measurements were around 50% for the three groups, without significant differences. CONCLUSIONS: This study shows that following sinus lift augmentation in sheep, MBCP()/fibrin grafts support new bone formation that is comparable to autografts, while providing better support for the dental implants. PMID- 19681971 TI - Timing of sirolimus conversion influences recovery of renal function in liver transplant recipients. AB - The long-term use of calcineurin inhibitors (CNI) leads to renal dysfunction in many liver transplant (LT) recipients. The purpose of this analysis is to evaluate renal function in patients converted from CNI to sirolimus (SRL). From May 2002-November 2006, 137 LT were performed in 125 patients, 72 of which were converted to SRL. Evaluation of SRL conversion was stratified by early conversion (<90 d from LT) (EC) vs. late conversion (LC). Renal function was evaluated using the six-point modification of diet in renal disease formula (estimated glomerular filtration rate [eGFR]). Forty-two patients on SRL and 40 on CNI had at least three months of follow-up and are included in the eGFR evaluation. At all time points after conversion, the EC group demonstrated a significantly higher mean eGFR than those in the LC group. A significant improvement in eGFR was seen within the EC group when comparing eGFR at time of conversion to eGFR at three, six, nine, and 12 months after conversion and last follow-up. The only improvement in the LC group was from conversion to the three-month time point. We conclude that EC to SRL results in a profound improvement in eGFR that begins at three months and is sustained beyond one yr. PMID- 19681972 TI - Do smaller adults wait longer for liver transplantation? A comparison of the UK and the USA data. AB - BACKGROUND: The number of patients on the UK and the USA liver transplant list is increasing. As size match is an important factor in the UK organ allocation, we studied the effect of recipient size on liver transplantation in the UK and the USA. METHODS: The UK Transplant and United Network for Organ Sharing databases were used to assess difference in access to transplantation between smaller adult patients and their larger counterparts over three time periods. Subsequently, proportions of split, NHBD and living-donor transplants were analyzed. RESULTS: There were 1576 UK and 29,150 USA patients in our analysis. The UK small patients have been significantly disadvantaged in access to transplantation particularly in early years and in adult only transplant units. This contrasts to the USA where smaller patients have never been disadvantaged and transplantation rates are steadily increasing. Split-liver transplants are being carried out in increasing numbers in the UK but not the USA. CONCLUSIONS: Small adults are still less likely to be transplanted at six months in adult only units in the UK. The lack of size matched organs for smaller adults and the overall decrease in rates of transplantation in the UK may be remedied by careful consideration of allocation policy and increased use of innovative techniques. PMID- 19681973 TI - Impact of genetic polymorphisms of the renin-angiotensin system and of non genetic factors on kidney transplant function--a single-center experience. AB - Renin-angiotensin-aldosterone system (RAAS) polymorphisms such as the angiotensinogen-gene-M235T-, the angiotensin-conversion enzyme (ACE)-gene I/D- and the angiotensin-II-type 1-receptor-(AT1R)-A1166C-polymorphism have been implicated in renal insufficiency and hypertension. We studied the association of these RAAS genotypes and non-genetic factors with transplant function and hypertension after renal graft transplantation (NTX). A total of 229 renal graft recipients, transplanted at a single center, were monitored up to 54 months and genotyped using polymerase chain reaction. The prevalence of the genotypes was comparable to a control group of healthy volunteers. Genotype and clinical outcome was analyzed using ANOVA, while the k-nearest neighbor method was used for a pattern recognition analysis of the complete database. Hypertension after NTX was not influenced by the RAAS polymorphisms. The DD-genotype of the ACE-I/D polymorphism was associated with significantly deteriorated renal transplant function during the months 18 to 30 after transplantation according to ANOVA at p < 0.05, as were non-genetic factors like long hospitalization, poor primary transplant function, and frequent rejections. Pattern recognition identified, the use of cyclosporine (odds ratio of 4.25) and the use of Ang II-receptor-blockers at discharge indicating the need of effective antihypertensive treatment (odds ratio of 3.26) as risk factors for transplant function loss. Altogether, the significant impact of the DD-genotype on the outcome after renal transplantation emphasizes the early identification of RAAS genotypes. PMID- 19681974 TI - Pancreas-after-kidney transplantation: to have and not have not. PMID- 19681975 TI - Higher tacrolimus trough levels on days 2-5 post-renal transplant are associated with reduced rates of acute rejection. AB - We analyzed the association between whole-blood trough tacrolimus (TAC) levels in the first days post-kidney transplant and acute cellular rejection (ACR) rates. Four hundred and sixty-four consecutive, deceased-donor kidney transplant recipients were included. All were treated with a combination of TAC, mycophenolate mofetil and prednisolone. Patients were analyzed in four groups based on quartiles of the mean TAC on days 2 and 5 post-transplant: Group 1: median TAC 11 ng/mL (n = 122, range 2-13.5 ng/mL), Group 2: median 17 ng/mL (n = 123, range 14-20 ng/mL), Group 3: median 24 ng/mL (n = 108, range 20.5-27 ng/mL) and Group 4: median 33.5 ng/mL (n = 116, range 27.5-77.5 ng/mL). A graded reduction in the rates of ACR was observed for each incremental days 2-5 TAC. The one-yr ACR rate was 24.03% (95% CI 17.26-32.88), 22.20% (95% CI 15.78-30.70), 13.41% (95% CI 8.15-21.63) and 8.69% (95% CI 4.77-15.55) for Groups 1-4, respectively (p = 0.003). This study suggests that higher early TACs are associated with reduced rates of ACR at one yr. PMID- 19681976 TI - Single versus dual renal transplantation from donors with significant arteriosclerosis on pre-implant biopsy. AB - BACKGROUND: Transplantation of kidneys from donor with arteriosclerosis seen on pre-implantation biopsy has not been well studied. METHODS: We retrospectively evaluated 20 dual kidney transplant (DKT) and 28 single (SKT) kidney transplant recipients with >or=12 months follow-up from donors with moderate arteriosclerosis (>or=25% luminal diameter narrowing). RESULTS: Death censored graft survival was 100% and 79%, respectively (p = 0.0339). DKT recipients had significantly lower mean creatinine levels at one, three, six, and nine months and spent somewhat less time on the waiting list (181 +/- 160 vs. 318 +/- 306 d, p = 0.1429). DKT patients received kidneys from significantly older donors (64 +/ 7 vs. 54 +/- 11 yr; p = 0.0012), proportionately more expanded criteria donors (95% vs. 54%; p = 0.0029), and more donors with hypertension (81% vs. 48%, p = 0.0344) and death related to cerebrovascular accident (100% vs. 71%, p = 0.0143); however, more DKT kidneys underwent machine perfusion (95% vs. 57%, p = 0.0068). Baseline recipient variables were comparable between the two groups including age, race, gender, retransplantation, and HLA mismatch. Pre-implant biopsy was notable for similar frequencies of moderate interstitial fibrosis (10% vs. 14%, respectively) and glomerulosclerosis. CONCLUSION: Among recipients of deceased donor kidneys with >25% arteriosclerosis, short-term outcomes after DKT were superior to that of SKT grafts. This approach may help to expand the donor-organ pool while optimizing outcomes. PMID- 19681977 TI - Usefulness of a program of neoplasia surveillance in liver transplantation. A preliminary report. AB - De novo malignancies are frequent complications after liver transplantation. Aim of the study is to evaluate whether a surveillance program for malignancy may improve patient survival. We have compared the survival after the diagnosis of malignancy (excluding cutaneous and hepatobiliary carcinomas and lymphoproliferative disease) of patients with symptomatic or incidental malignancies with patients with neoplasia diagnosed on screening. Two hundred and eighty patients with a follow-up greater than three months were followed for a median of 77.5 months (total follow-up: 1515 patient-yr). Thirty-three patients developed 41 malignancies. When compared with general population, the entire cohort of liver transplant recipients had a significantly higher risk of malignancy (relative risk: 2.34), gastrointestinal tract (relative risk: 2.52), urological tract (relative risk: 2.94) and head and neck cancer (relative risk: 4.14), and cancer-related death (relative risk: 2.35). All nine patients diagnosed with cancer with active screening are currently alive and free of malignancy after a median follow-up of 25 months. By contrast, 18/24 patients with diagnosis of cancer prompted by symptoms or incidentally diagnosed died as a consequence of the cancer (median survival: 13.5 months). The difference in survival between both groups was significant (p = 0.002). In conclusion, a close surveillance protocol for the diagnosis of malignancy could be life-saving in liver transplant recipients. PMID- 19681978 TI - Donor transmission of malignant melanoma to a liver graft recipient: case report and literature review. AB - Post-transplant malignancy of donor origin is a rare complication of organ transplantation, most likely transmitted as micrometastases within the parenchyma of the donor organ or from circulating tumor cells contained within the organ. Patient survival is dependent upon early diagnoses, and differentiation of the malignancy as of donor or recipient derivation is important in developing a treatment modality. The utilization of fluorescent in situ hybridization chromosome analysis and DNA sequence analysis of the tumor cells can assist in this determination. This case report describes the management of donor transmitted malignant melanoma in a liver graft recipient and a review of the current literature. PMID- 19681979 TI - Intraoperative assessment of microperfusion with visible light spectroscopy for prediction of anastomotic leakage in colorectal anastomoses. AB - PURPOSE: Anastomotic leakage is associated with increased morbidity and mortality. However, there is no accurate tool to predict its occurrence. We evaluated the predictive value of visible light spectroscopy (VLS), a novel method to measure tissue oxygenation [saturated O(2) (StO(2) )], for anastomotic leakage of the colon and the rectum. METHOD: Oxygen saturation in the bowel was measured in 77 colorectal resections. The anastomosis was between 2 and 30 cm (mean 13 cm) from the anal verge. The oxygen saturation was measured in the colon and rectum before and after anastomosis construction. This was compared with a reference measurement in the caecum. Data on postoperative complications were prospectively collected. RESULTS: Anastomotic leakage occurred in 14 (18%) patients. When compared with a leaking anastomosis, normal anastomoses showed rising O(2) values during the operation (mean StO(2) 72.1 +/- 9.0-76.7 +/- 8.0 vs 73.9 +/- 7.9-73.1 +/- 7.4) (P <= 0.05). There were also higher StO(2) values in the caecum compared with those which ultimately leaked (73.6 +/- 5.7 normal anastomoses, 69.6 +/- 5.6 anastomotic leaks) (P <= 0.05). Both StO(2) values were predictive of anastomotic leakage. CONCLUSION: Tissue oxygenation O(2) appears to be a potentially useful means of predicting anastomotic leakage after colorectal anastomosis. PMID- 19681980 TI - A novel two-dimensional dynamic anal ultrasonography technique to assess anismus comparing with three-dimensional echodefecography. AB - OBJECTIVE: The aim of this prospective study was to test two-dimensional dynamic anorectal ultrasonography (2D-DAUS) in the assessment of anismus and compare it with echodefecography (ECD). METHOD: Fifty consecutive female patients with outlet delay were submitted to 2D and 3D-DAUS, measuring the relaxing or contracting puborectalis muscle angle during straining. The patients were assigned to one of two groups based on ECD findings. Group I consisted of 29 patients without anismus and group II included 21 patients diagnosed with anismus. Subsequently 2D-DAUS images were checked for anismus and compared with ECD findings. RESULTS: Upon straining, the angle produced by the movement of the puborectalis muscle decreased in 26 out of the 29 (89.6%) patients of group I and increased 19 out of the 21 (90.4%) patients of group II. The mean angle during straining differed significantly between group I and group II. The index of agreement between the two scanning modes was 89.6% (26/29) for group I (Kappa: 0.796; CI: 95%; range: 0.51-1.0) and 90.4% (19/21) for group II (Kappa: 0.796; CI: 95%; range: 0.51-1.0). CONCLUSION: Two-dimensional dynamic anal ultrasonography can be used as an alternative method to assess patients with anismus, although the 3-D modality is more precise to evaluate the PR angle as the sphincters integrity as the whole muscle length is clearly visualized. PMID- 19681981 TI - Use of Echelon 60 ENDOPATH stapling device in open ileal pouch anal anastomosis (IPAA) surgery. PMID- 19681982 TI - Chewing function impacts oral health-related quality of life among institutionalized and community-dwelling Korean elders. AB - OBJECTIVE: The aim of this study was to assess the association of chewing ability to oral health-related quality of life (OHRQoL) measured by the Oral Health Impact Profile-14 (OHIP-14) controlling for clinical oral health status, self reported health status, demographic factors, and socioeconomic conditions among community-dwelling and institutionalized Korean elders. METHODS: This cross sectional study comprised a sample of 307 community-dwelling and 102 institutionalized people over the age of 60, using a cluster sampling procedure. A questionnaire was implemented and a clinical oral examination was completed for each subject. The outcome variable of interest was the OHIP-14 score, and its associations with chewing ability, objective oral health status, self-reported health status, demographic factors, and socioeconomic conditions were assessed. Because of highly-skewed distribution of the OHIP-14 scores, nonparametric analytic methods were used. The final model was developed using a multivariable two-level logistic regression model for a dichotomized OHIP-14 score to account for the cluster sampling method applied to this study. RESULTS: The mean age of the participants was 75.4 years, with 67.7% being women. The median OHIP-14 score was 7. Negative oral health impacts were experienced fairly often or very often by a total of 36.4% of elderly. In the final model, elders who could chew none to three and four to six foods among seven indicator foods were 3.4 (P = 0.010) and 2.0 (P = 0.040) times more likely, respectively, to have worse OHRQoL compared with elders who could chew all seven food types. Also significant associations with worse OHRQoL were shown for being concerned about oral health [Odds Ratio (OR) = 4.9, P = 0.002], fair or better self-reported oral health (OR = 0.12, P = 0.002), very good/good self-reported general health (OR = 0.38, P = 0.008), being married (OR = 2.0, P = 0.054), and having a favourable economic status (OR = 0.43, P = 0.042). CONCLUSIONS: This study showed highly significant association between chewing ability and OHRQoL measured by the OHIP-14 score after controlling for related factors. Amelioration of chewing ability might independently contribute to improving the OHRQoL of elders. PMID- 19681983 TI - Evaluation of the mechanical and chemical control of dental biofilm in patients with Down syndrome. AB - OBJECTIVE: To evaluate the mechanical and chemical control of dental biofilm in patients with Down syndrome, using different experimental dentifrices. MATERIAL AND METHODS: Forty institutionalized children between ages 7 and 13 years in the mixed dentition phase participated in this study. An experimental cross-over, blind clinical trial was used, having the following protocols: fluoridated dentifrice (protocol G1); fluoridated dentifrice + chlorhexidine (protocol G2); fluoridated dentifrice + chlorhexidine + plaque-disclosing agent (protocol G3); and fluoridated dentifrice + plaque-disclosing agent (protocol G4). Each experimental stage lasted 10 days with a 15-day washout. The evaluated parameters were Plaque Index and gingival bleeding. RESULTS: The initial clinical conditions between each stage were similar. Statistical differences were observed (P < 0.001) for the clinical conditions evaluated before and after the treatments. The dentifrices containing plaque-disclosing agent, irrespective of their association with chlorhexidine, produced a greater reduction in the final plaque index. As for gingival bleeding, the dentifrice containing erythrosine and the one containing chlorhexidine produced similar results. The dentifrice containing an association of chlorhexidine and erythrosine gave the best results. CONCLUSION: With the methodology employed, it was possible to conclude that the combination of drugs (chlorhexidine, fluorine and erythrosine) within one dentifrice can be useful in controlling dental biofilm and in the reduction of gingival bleeding. PMID- 19681984 TI - Reproducibility and accuracy of the ICDAS-II for occlusal caries detection. AB - OBJECTIVES: The aim of this in vitro study was to assess the inter- and intra examiner reproducibility and the accuracy of the International Caries Detection and Assessment System-II (ICDAS-II) in detecting occlusal caries. METHODS: One hundred and sixty-three molars were independently assessed twice by two experienced dentists using the 0- to 6-graded ICDAS-II. The teeth were histologically prepared and classified using two different histological systems [Ekstrand et al. (1997) Caries Research vol. 31, pp. 224-231; Lussi et al. (1999) Caries Research vol. 33, pp. 261-266] and assessed for caries extension. Sensitivity, specificity, accuracy and area under the ROC curve (A(z)) were obtained at D(2) and D(3) thresholds. Unweighted kappa coefficient was used to assess inter- and intra-examiner reproducibility. RESULTS: For the Ekstrand et al. histological classification the sensitivity was 0.99 and 1.00, specificity 1.00 and 0.69 and accuracy 0.99 and 0.76 at D(2) and D(3), respectively. For the Lussi et al. histological classification the sensitivity was 0.91 and 0.75, specificity 0.47 and 0.62 and accuracy 0.86 and 0.68 at D(2) and D(3), respectively. The A(z) varied from 0.54 to 0.73. The inter- and intra-examiner kappa values were 0.51 and 0.58, respectively. CONCLUSIONS: ICDAS-II presented good reproducibility and accuracy in detecting occlusal caries, especially caries lesions in the outer half of the enamel. PMID- 19681986 TI - Value of artificial habitats for amphibian reproduction in altered landscapes. AB - Installation and maintenance of stormwater ponds to detain and treat runoff from impervious surfaces is a common method of stormwater control in developed areas. That these ponds capture pollutants, however, is of concern for wildlife species that use the ponds, particularly pond-breeding amphibians. To assess the relative contribution of stormwater ponds to the persistence of amphibian populations in suburban landscapes, we compared amphibian use of stormwater ponds and other available wetlands in suburban and forested watersheds. We surveyed three suburban and three primarily forested first-order watersheds to identify all potential wetlands that might serve as breeding sites for pond-breeding amphibians. We performed call, egg-mass, and larval surveys to measure breeding effort at each wetland in spring and summer 2007 and 2008. In suburban watersheds most (89%) of the wetlands that had breeding activity were either stormwater ponds or otherwise artificial. This pattern was also evident in the forested watersheds, where amphibians were primarily found breeding in wetlands created by past human activity. Late-stage larvae were found only in anthropogenic wetlands in all study areas because the remaining natural wetlands did not hold water long enough for larvae to complete development. Our results suggest that in urban and suburban landscapes with naturally low densities of wetlands, wetlands created by current or historic land uses may be as important to amphibian conservation as natural wetlands or pools and that management strategies directed at urban and suburban landscapes should recognize and incorporate human-created wetlands. PMID- 19681985 TI - Linking mother access to dental care and child oral health. AB - OBJECTIVES: The authors examined whether low-income mothers, who have a regular source of dental care (RSDC), rate the dental health of their young children higher than mothers without an RSDC. METHODS: From a population of 108,151 children enrolled in Medicaid aged 3 to 6 years and their low-income mothers in Washington state, a disproportionate stratified random sample of 11,305 children aged 3 to 6 years was selected from enrollment records in four racial/ethnic groups: 3791 Black; 2806 Hispanic; 1902 White; and 2806 other racial/ethnic groups. A mixed-mode survey was conducted to measure mother RSDC and mother ratings of child's dental health and pain. The unadjusted response rate was 44%, yielding the following eligible mothers: 816 Black, 1309 Hispanic, 1379 White, 237 Asian, and 133 American-Indian. Separate regression models for Black, Hispanic, and White mothers estimated associations between the mothers having an RSDC and ratings of child dental health. RESULTS: Across racial/ethnic groups, mothers with an RSDC consistently rated their children's dental health 0.15 higher on a 1-to-5 scale (where '1' means 'poor' and '5' means 'excellent') than mothers without an RSDC, controlling for child and mother characteristics and the mothers' propensity to have an RSDC. This difference can be interpreted as a net movement of one level up the scale by 15% of the population. CONCLUSIONS: Across racial/ethnic groups, low-income mothers who have a regular source of dental care rate the dental health of their young children higher than mothers without an RSDC. PMID- 19681987 TI - Effects of reduced-impact logging on fish assemblages in central Amazonia. AB - In Amazonia reduced-impact logging, which is meant to reduce environmental disturbance by controlling stem-fall directions and minimizing construction of access roads, has been applied to large areas containing thousands of streams. We investigated the effects of reduced-impact logging on environmental variables and the composition of fish in forest streams in a commercial logging concession in central Amazonia, Amazonas State, Brazil. To evaluate short-term effects, we sampled 11 streams before and after logging in one harvest area. We evaluated medium-term effects by comparing streams in 11 harvest areas logged 1-8 years before the study with control streams in adjacent areas. Each sampling unit was a 50-m stream section. The tetras Pyrrhulina brevis and Hemigrammus cf. pretoensis had higher abundances in plots logged > or =3 years before compared with plots logged <3 years before. The South American darter (Microcharacidium eleotrioides) was less abundant in logged plots than in control plots. In the short term, the overall fish composition did not differ two months before and immediately after reduced-impact logging. Temperature and pH varied before and after logging, but those differences were compatible with normal seasonal variation. In the medium term, temperature and cover of logs were lower in logged plots. Differences in ordination scores on the basis of relative fish abundance between streams in control and logged areas changed with time since logging, mainly because some common species increased in abundance after logging. There was no evidence of species loss from the logging concession, but differences in log cover and ordination scores derived from relative abundance of fish species persisted even after 8 years. For Amazonian streams, reduced-impact logging appears to be a viable alternative to clear-cut practices, which severely affect aquatic communities. Nevertheless, detailed studies are necessary to evaluated subtle long-term effects. PMID- 19681988 TI - Successful treatment of severe keratosis pilaris rubra with a 595-nm pulsed dye laser. PMID- 19681989 TI - Liporhinophyma: an unusual presentation of rhinophyma. PMID- 19681990 TI - Cutaneous Desmoid tumor: resolution of the surgical defect with a dermal regeneration template and an epidermal autograft. PMID- 19681991 TI - Photodynamic therapy for patients with Basal cell nevus syndrome. PMID- 19681992 TI - Primary cutaneous rhabdomyosarcoma in an adult. PMID- 19681993 TI - Management of carcinoma of the skin in solid organ transplant recipients with oral capecitabine. PMID- 19681994 TI - Treatment of cutaneous squamous cell carcinoma with perineural invasion using Mohs micrographic surgery: report of two cases and review of the literature. PMID- 19681995 TI - Facilitated multiple intradermal injections using the double-syringe technique. PMID- 19681996 TI - Scalp myiasis associated with advanced basal cell carcinoma. PMID- 19681998 TI - Split-skin grafting from the scalp: the hidden advantage. AB - BACKGROUND: Split-skin grafting is a routine reconstructive technique associated with large variation in practice. Grafts from the thigh, buttock, or abdomen take a long time to heal and may leave unpleasant, hypopigmented scars. Retrospective reports favor the scalp as a donor site in burn patients. OBJECTIVE: Evaluation of duration of healing, cosmetic outcome, and safety of split-skin grafting from the scalp in patients receiving dermatologic surgery. MATERIAL AND METHODS: One hundred sixty-six consecutive patients (85 men, 81 women) were treated for coverage of chronic leg ulcers or other large skin defects with a split-skin graft taken from the posterior scalp. Area and thickness of the graft, healing time, and adverse events were documented. RESULTS: Mean healing time until complete reepithelization was 5.4+/-1.0 days for a single harvest (median 5 days). No major complications occurred. Spotted alopecia was a rare event. Almost all (96.5%) of the patients would undergo split-skin harvesting from the occipital scalp again if needed. CONCLUSIONS: Advantages of the scalp as a donor site include rapidity of wound healing, low risk of complications, and excellent cosmetic results. The large number of hair follicles containing the epidermal stem cell pool can explain these advantages. PMID- 19682000 TI - Treatment of angiofibromas of tuberous sclerosis with 5-aminolevulinic acid blue light photodynamic therapy followed by immediate pulsed dye laser. PMID- 19681999 TI - Enhanced efficacy of photodynamic therapy with methyl 5-aminolevulinic acid in recalcitrant periungual warts after ablative carbon dioxide fractional laser: a pilot study. AB - BACKGROUND: Photodynamic therapy (PDT) with aminolevulinic acid (ALA) has been used to improve recalcitrant periungual warts, but most lesions achieved complete remission after more than four sessions, and some lesions did not respond to the method. In this pilot study, the potential for synergistic effects of the combination of ablative carbon dioxide (CO(2)) fractional laser and methyl 5-ALA (MAL)-PDT for the treatment of recalcitrant periungual warts was examined. MATERIALS AND METHODS: Twelve Korean patients (8 women and 4 men aged 20-45, mean age 27.9) with a total of 40 periungual warts were enrolled in the present study. The lesions were treated using an ablative CO(2) fractional laser. Immediately after each fractional treatment, MAL was applied on the periungual warts, and 3 hours later, such areas were illuminated with a red light at a dose of 50 J/cm(2) for 15 minutes. RESULTS: After a mean of 2.2 treatments per wart, a mean clearance of 100% was achieved in 36 (90%) warts. Two warts (5%) had 50% clearance, and two (5%) showed no response after three treatments. There were no recurrences of the warts that had achieved 100% clearance during the follow-up period of 6 months. Most of the treatments had no severe side effects during or after their administration. CONCLUSION> A potential for enhanced clinical results when using combined ablative CO(2) fractional laser and MAL-PDT for the treatment of periungual warts was shown in this pilot study. PMID- 19682001 TI - Fractional photothermolysis for the treatment of postinflammatory hyperpigmentation. PMID- 19682002 TI - Earthquake mortality in Pakistan. AB - A strong earthquake, measuring 7.6 on the Richter scale, hit northern Pakistan on 8 October 2005, causing massive destruction, including an official death toll of 73,276. Four cross-sectional surveys were performed in late 2005 to assess mortality before the event, on the day, and subsequently. Two surveys were community-based and two were situated in camps for internally displaced persons. Crude mortality rates were low in the 3.5 months preceding the earthquake (less than 0.1 deaths per 10,000 per day) and slightly higher in the six-to-eight weeks after the earthquake (ranging from 0.10-0.43 per 10,000 per day). On 8 October 2005, approximately two per cent of the population in one community survey died and around five per cent in the other three surveys. Children less than five years and adults more than or equal to 50 years tended to have a higher risk of mortality on the day of the disaster. These results corroborate the high mortality caused by the earthquake. PMID- 19682003 TI - Mortality rate and confidence interval estimation in humanitarian emergencies. AB - Surveys are conducted frequently in humanitarian emergencies to assess the health status of the population. Most often, they employ complex sample designs, such as cluster sampling. Mortality is an indicator commonly estimated in such surveys. Confidence limits provide information on the precision of the estimate and it is important to ensure that confidence limits for a mortality rate account for the survey design and utilise an acceptable methodology. This paper describes the calculation of confidence limits for mortality rates from surveys using complex sampling designs and a variety of software programmes and methods. It contains an example that makes use of the SAS, SPSS, and Epi Info software programmes. Of the three confidence interval methods examined--the ratio command approach, the modified rate approach, and the modified proportion approach--the paper recommends the ratio command approach to estimate mortality rates with confidence limits. PMID- 19682004 TI - Emergency medical preparedness during the 2006 World Cup in Frankfurt, Germany. AB - This paper describes emergency medical preparedness during FIFA (Federation Internationale de Football Association) World Cup matches in Frankfurt, Germany, in 2006. The methods employed were document analysis and personal observation of games over five days in June-July 2006. The medical authorities in Frankfurt drew on a wide range of scientific literature and experiences to elaborate a National Concept. They paid attention to different models of handling mass catastrophes in shaping the final version of the document. The participation of designated authorities, associations, and volunteer organisations was coordinated sufficiently and the games in Frankfurt proceeded without great incident, even though more than 300,000 people in total attended. The adopted emergency medical procedure was appropriate for a mass gathering event. Official and volunteer organisations collaborated precisely in emergency preparedness. While one uniform concept for all mass gatherings events cannot be developed, case reports and experiences are useful tools. PMID- 19682005 TI - Role of information in collective action in dynamic disaster environments. AB - This study explores the major issues facing collective response operations after destructive earthquakes. The small-n case study design employs qualitative and quantitative methods to investigate the decision-making process in a context of seismic risk to exemplify how public managers can utilise information and communication systems to ensure collaborative actions in managing an extreme event. Fifty-eight semi-structured interviews with 39 key decision-makers and researchers and content analyses of daily reports from Cumhuriyet comprise the main data sources. The study compares and contrasts the Turkish disaster management system following the Marmara and Duzce earthquakes of 1999. It addresses whether the use of information and communication technologies significantly affected its performance. The study's findings reveal that difficulties in accessing and exchanging timely and accurate disaster-relevant information inhibited coordination during the Marmara response while increased communication functions improved coordination and search-and-rescue activities during the Duzce response. PMID- 19682006 TI - The 9/21 earthquake in Taiwan: a local government disaster rescue system. AB - This paper employs a three-element model to examine how the disaster rescue system of the government of Nantou County in Middle Taiwan functioned following the earthquake of 21 September 1999. The three elements are information gathering, local government mobilisation, and inter-organisational cooperation. The paper finds that the Nantou County government needs to address many problems associated with these three elements. Disaster information, for example, was not processed instantly because of the destruction of the electricity and telephone systems in the earthquake. Insufficient information caused ineffectiveness in the realms of mobilisation and inter-organisational cooperation. As for mobilisation, while the Nantou County magistrate successfully used specific information to encourage flows of huge resources in the county, he did not successfully mobilise human resources there. With regard to inter-organisational cooperation, myriad voluntary actors and international rescue teams travelled to Nantou County, but the fire and police services experienced cooperation and coordination problems. PMID- 19682007 TI - Are treelines advancing? A global meta-analysis of treeline response to climate warming. AB - Treelines are temperature sensitive transition zones that are expected to respond to climate warming by advancing beyond their current position. Response to climate warming over the last century, however, has been mixed, with some treelines showing evidence of recruitment at higher altitudes and/or latitudes (advance) whereas others reveal no marked change in the upper limit of tree establishment. To explore this variation, we analysed a global dataset of 166 sites for which treeline dynamics had been recorded since 1900 AD. Advance was recorded at 52% of sites with only 1% reporting treeline recession. Treelines that experienced strong winter warming were more likely to have advanced, and treelines with a diffuse form were more likely to have advanced than those with an abrupt or krummholz form. Diffuse treelines may be more responsive to warming because they are more strongly growth limited, whereas other treeline forms may be subject to additional constraints. PMID- 19682008 TI - Diagnosing pulmonary embolism in the emergency department: maybe we do matter, after all. PMID- 19682009 TI - Review article: Emergency Department implications of the TASER. AB - The TASER is a conducted electricity device currently being introduced to the Australian and New Zealand police forces as an alternative to firearms in dealing with violent and dangerous individuals. It incapacitates the subject by delivering rapid pulses of electricity causing involuntary muscle contraction and pain. The use of this device might lead to cardiovascular, respiratory, biochemical, obstetric, ocular and traumatic sequelae. This article will summarize the current literature and propose assessment and management recommendations to guide emergency physicians who will be required to review these patients. PMID- 19682010 TI - Review article: management of acute severe and near-fatal asthma. AB - Despite a decline in the Australian overall asthma mortality, near-fatal/critical asthma continues to be a significant management issue for emergency physicians and intensivists. Near-fatal asthma is a unique subtype of asthma, with a variety of clinical presentations, requiring rapid and aggressive intervention. The pharmacological and non-pharmacological management of near-fatal asthma remains very complex. The present review discusses recent advances and evidence for current available strategies targeting this time critical emergency. PMID- 19682011 TI - Emergency Department diagnosis of pulmonary embolism is associated with significantly reduced mortality: a linked data population study. AB - OBJECTIVES: We characterized patients admitted via ED with a principal hospital discharge diagnosis of pulmonary embolism (PE) and compared mortality of those diagnosed in the ED with those diagnosed after admission. METHODS: Patients with a hospital discharge diagnosis ICD 10 I26 presenting to the ED in Perth, Western Australia between 1 July 2000 and 30 December 2006 had records from the Emergency Department Information System linked to the Western Australian Hospital Morbidity Data System and the death registry. RESULTS: Of 2250 patients (mean age 60.4), 1227 (54.5%) were female. Of 1931 patients with an ED diagnosis recorded, 1207 (62.5%) were diagnosed with PE in ED. Of these, 383 (17.0%) had presented to an ED within 28 days previously, 142 (37.1%) with either chest pain or breathing problems, with 207 (54.0%) admitted but not receiving a principal hospital discharge diagnosis of PE. There were 127 (5.6%) in-hospital deaths. Controlling for age and comorbidity with logistic regression, patients diagnosed with PE in ED were less likely to die in hospital, within 7 and 30 days of ED arrival, than those diagnosed after admission (adjusted OR 0.31, 95% CI 0.20-0.47; adjusted OR 0.32, 95% CI 0.19-0.53; adjusted OR 0.30, 95% CI 0.20-0.44; respectively). CONCLUSION: Making the diagnosis of PE in ED was associated with a substantial survival advantage that persisted after hospital discharge. PMID- 19682012 TI - Factors influencing consistency of triage using the Australasian Triage Scale: implications for guideline development. AB - OBJECTIVE: To examine the influence of the nurse, the type of patient presentation and the level of hospital service on consistency of triage using the Australasian Triage Scale. METHODS: A secondary analysis of survey data was conducted. The main study was undertaken to measure the reliability of 237 scenarios for inclusion in a national training programme. Nurses were recruited from a quota sample of Australian ED according to peer group. Analysis was performed to determine concordance: the percentage of responses in the modal triage category. Analysis of variance (anova) and Pearson correlations were used to investigate associations between the explanatory variables and concordance. RESULTS: A total of 42/50 (84%) participants returned questionnaires, providing 9946 scenario responses for analysis. Significant differences in concordance were observed by variables describing the type of patient presentation and level of urgency. Mean scores for the comparison group (adult pain; 70.7%) were higher than the groups involving a mental health or pregnancy presentations (61.4%; P15 degrees dorsal angulation were classified as 'displaced'. MAIN OUTCOME MEASURES: Radiographic analysis of the dorsal angle on post-reduction and 6 week post-fracture X-rays. A combined end-point of poor outcome, as defined by either poor radiological result and/or progression to surgery. RESULTS: In the group of patients with displaced fractures, 69 of 114 (61%) went on to have an operation or a poor radiographic outcome versus 8 of 48 (17%) in the group with minimally displaced fractures, an absolute difference of 44% (95% CI 30-57%). Patients who had a minimally displaced fracture with an adequate reduction went on to have a satisfactory 6 week X-ray in 37 of 43 cases (86%; 95% CI 75-96%). Patients who had a displaced fracture and an adequate reduction had a satisfactory 6 week X-ray in 42 of 86 cases (49%; 95% CI 38-59%). Patients who had a displaced fracture and an inadequate reduction had a satisfactory 6 week X-ray in only 3 of 22 cases (14%; 95% CI 0-28%). CONCLUSION: The study highlights the importance of the initial 'on arrival' and 'post-reduction' X-rays in the ED. Displaced fractures are more likely to go onto poor outcome, as are inadequately reduced fractures. Medical officers working in ED should be aware of the importance of measuring the dorsal angle. They should be referring patients with >15 degrees dorsal angulation to orthopaedics early. Reduction should not be accepted until the dorsal angle has been adequately corrected. PMID- 19682016 TI - Emergency Department access block occupancy predicts delay to surgery in patients with fractured neck of femur. AB - OBJECTIVE: The present study aimed to identify any relationship between existing access block occupancy (ABO) at the time of patient presentation and delay to definitive procedure. METHODS: Retrospective descriptive cohort study of all patients aged over 50 years with an ED diagnosis of fractured neck of femur admitted through a tertiary ED over 2 years. The independent variable was the ABO at the start of the hour in which the patient presented, derived from existing ED records, and expressed as the quartile for that hour of the day. The dependent variable was start of surgery more than 24 h after arrival without a documented reason for delay. The data abstractor was blinded to the ABO. RESULTS: All 442 diagnoses of fractured neck of femur recorded in the ED were reviewed, 73 were excluded (16 age, 5 misdiagnosis, 31 no surgery, 21 documented medical reasons for delay). There was a significant relationship between ABO quartile and the rate of delay to surgery ranging from 54% (95% CI 43-63%) for those presenting in the lowest ABO quartile to 77% (68-85%) in the highest (P= 0.006, chi(2)). Subgroup analysis showed that arrival ABO predicted patient access block, and that patient access block was associated with delay to surgery and longer postoperative length of stay (geometric mean 12.9 vs 9.9 days, P < 0.01, t-test). CONCLUSIONS: The number of access block patients at the time of arrival directly predicts delay to operation in this setting. This suggests that access block occupancy is a marker of hospital dysfunction. PMID- 19682017 TI - 27 years of croup: an update highlighting the effectiveness of 0.15 mg/kg of dexamethasone. AB - OBJECTIVE: To update an earlier observational study (1980-1995) documenting dramatic improvements in the management of croup with the mandatory use of a single oral dose of dexamethasone and to ascertain whether a reduction from a dose of 0.6 to 0.15 mg/kg in 1995 maintained these improved outcomes over the next 11 years. METHODS: We evaluated retrospectively the experience of children with croup in Princess Margaret Hospital for Children, the only tertiary paediatric hospital in Western Australia, over the subsequent 11 year period from 1996 to 2006 inclusive. Data were updated from ED, general hospital and the intensive care unit records to show the numbers of children presenting to the hospital, admitted, transferred to intensive care and intubated. We also recorded the length of hospital stay and representation rate of all cases within 7 days. RESULTS: The dramatic improvements in outcomes for croup, including reduced admission rates, length of stay, transfers to the intensive care unit, intensive care unit days and number of intubations as reported in our earlier paper, were maintained using 0.15 mg/kg dexamethasone. Admission rates for croup have fallen from 30% in the early 1990s to less than 15% in recent years, whereas the representation rate has risen slightly. CONCLUSION: The improved outcomes for children with croup presenting to our paediatric ED have been maintained with a reduced, single oral dose of 0.15 mg/kg of dexamethasone. PMID- 19682018 TI - What is the nature of the emergence phenomenon when using intravenous or intramuscular ketamine for paediatric procedural sedation? AB - OBJECTIVE: Ketamine has become the drug most favoured by emergency physicians for sedation of children in the ED. Some emergency physicians do not use ketamine for paediatric procedural sedation (PPS) because of concern about emergence delirium on recovery. The present study set out to determine the true incidence and nature of this phenomenon. METHODS: Prospective data relating to any emergence agitation, crying, hallucinations, dreams, altered perceptions, delirium and necessary interventions were recorded in consecutive cases of ketamine PPS from March 2002 to June 2007, and analysed. Standard inclusion and exclusion criteria for the use of ketamine were followed. RESULTS: A total of 745 prospective data collection records were available for analysis over the 5 year period. Of all, 93 (12.5%) children cried on awakening when recovering from PPS, 291 (39%) experienced pleasant altered perceptions and 16 (2.1%) experienced what was called 'emergence delirium'. None required any active treatment and all except one settled within 20 min. There was no evidence of an increased rate of nightmares on telephone follow up in the weeks post procedure. CONCLUSION: The belief that ketamine, in the doses used for ED PPS, causes frequent emergence delirium is flawed. A pleasant emergence phenomenon is common, but is not distressing for the child, and has no long-term (up to 30 days) negative sequelae. Rarely, there is anxiety or distress on awakening from ketamine sedation, which settles spontaneously. This should not deter emergency physicians from using ketamine for PPS. PMID- 19682019 TI - Factors influencing rural versus metropolitan work choices for emergency physicians. AB - OBJECTIVES: To survey Fellows of the Australasian College for Emergency Medicine (FACEM) on how a range of factors influenced their decision to accept their most recent position. To compare this information between rural and metropolitan FACEM. METHODS: Analytical cross-sectional survey of FACEM. Sections included baseline demographics and a range of questions regarding the presence, absence and influence of 14 professional and 12 personal/external factors on the decision to accept their current position. RESULTS: Questionnaires were returned by 498 (61.9%) of 805 FACEM. Eighty-seven (18.4%) were currently employed in rural areas. Rural FACEM were more likely to be male (odds ratio 2.0 [95% CI 1.1-3.9]) and to have worked for >12 months as a registrar in a rural hospital (odds ratio 4.5 [95% CI 2.2-9.1]). Negative influences for FACEM accepting rural positions included lack of access to continuing education, less acceptable on-call arrangements, fewer employment opportunities for their partner and less educational opportunities for their children. Positive influences included acceptable remuneration, desirable lifestyle, a higher indigenous caseload and more affordable housing. CONCLUSIONS: The influence of different types of factors appears to differ between rural and metropolitan FACEM and this information might assist in the formulation of strategies aimed at increasing the rural workforce. PMID- 19682020 TI - Atrio-oesophageal fistula: an emergent complication of radiofrequency ablation. AB - A 72-year-old presented with features of sepsis, neurological sequelae and chest pain after a radiofrequency ablation for AF. Chest CT scan revealed a life threatening condition not previously reported in emergency medicine journals. PMID- 19682021 TI - Treatment of migraine in Australian Emergency Departments. PMID- 19682022 TI - Intranasal fentanyl paediatric clinical practice guidelines. PMID- 19682024 TI - The metabolic syndrome in overweight epileptic patients treated with valproic acid. AB - PURPOSE: To evaluate the presence of metabolic syndrome (MS) in children and adolescents treated with valproate (VPA). METHODS: One hundred fourteen patients (54 male and 60 female) were studied. These patients were followed from the beginning of therapy for at least 24 months; at the end of follow-up, 46 patients (40.4%) had a considerable increase in body weight, whereas the other patients (59.6%) remained with the same weight. The MS was defined as having at least three of the following: abdominal obesity, dyslipidemia, glucose intolerance, and hypertension. RESULTS: Forty-six patients developed obesity; 20 (43.5%) of 46 patients developed MS. Abnormal glucose homeostasis was identified in 45% of patients. High total serum cholesterol concentrations were noted in 10 (50%), high serum triglyceride concentrations in 7 (35%), and low high-density lipoprotein (HDL) in 15 (75%) of the 20 subjects with MS. However, there were no significant differences in the features of MS between boys and girls with MS. CONCLUSIONS: Patients who gain weight during VPA therapy can develop MS with a possible risk of cardiovascular disease. PMID- 19682026 TI - A new rapid micromethod for the assay of phenobarbital from dried blood spots by LC-tandem mass spectrometry. AB - Advantages of dried blood spot include low invasiveness, ease and low cost of sample collection, transport, and storage. We used tandem mass spectrometry (LC MS/MS) to determine phenobarbital levels on dried blood spot specimens and compared this methodology to commercially available particle enhanced turbidimetric inhibition immunoassay (PETINIA) in plasma/serum samples. The calibration curve in matrix using D(5)-phenobarbital as internal standard was linear in the phenobarbital concentration range of 1-100 mg/L (correlation coefficient 0.9996). The coefficients of variation in blood spots ranged 2.29 6.71% and the accuracy ranged 96.54-103.87%. There were no significant differences between the concentrations measured using PETINA and LC-MS/MS (both had similar precision and accuracy) however, LC-MS/MS allows at least 1.5 times higher throughput of phenobarbital analysis and additionally offers ease of sample collection which is particularly important for newborns or small infants. PMID- 19682025 TI - Treatment of early and late kainic acid-induced status epilepticus with the noncompetitive AMPA receptor antagonist GYKI 52466. AB - PURPOSE: Benzodiazepines such as diazepam may fail to effectively treat status epilepticus because benzodiazepine-sensitive GABA(A) receptors are progressively internalized with continued seizure activity. Ionotropic glutamate receptors, including AMPA receptors, are externalized, so that AMPA receptor antagonists, which are broad-spectrum anticonvulsants, could be more effective treatments for status epilepticus. We assessed the ability of the noncompetitive AMPA receptor antagonist GYKI 52466 to protect against kainic acid-induced status epilepticus in mice. METHODS: Groups of animals treated with kainic acid received GYKI 52466 (50 mg/kg followed in 15 min by 50 mg/kg) or diazepam (25 mg/kg followed in 20 min by 12.5 mg/kg) beginning at 5 min of continuous seizure activity or 25 min later. The duration of seizure activity was determined by EEG recording from epidural cortical electrodes. RESULTS: Both GYKI 52466 and diazepam rapidly terminated electrographic and behavioral seizures when administered early. However, diazepam-treated animals exhibited more seizure recurrences. With late administration, GYKI 52466 also rapidly terminated seizures and they seldom recurred, whereas diazepam was slow to produce seizure control and recurrences were common. Although both treatments caused sedation, GYKI 52466-treated animals retained neurological responsiveness whereas diazepam-treated animals did not. GYKI 52466 did not affect blood pressure whereas diazepam caused a sustained drop in mean arterial pressure. DISCUSSION: Noncompetitive AMPA receptor antagonists represent a promising approach for early treatment of status epilepticus; they may also be effective at later times when there is refractoriness to benzodiazepines. PMID- 19682028 TI - Vagus nerve stimulation induces changes in respiratory sinus arrhythmia of epileptic children during sleep. AB - PURPOSE: This study analyzed the direct short-term effect of vagus nerve stimulation (VNS) on respiratory sinus arrhythmia (RSA) in children with pharmacoresistant epilepsy. METHODS: RSA magnitude is calculated as the ratio between maximum and minimum heart rate for each respiratory cycle-before, during, and after the actual VNS period. In 10 children, changes in RSA magnitude were evaluated on polysomnographic recordings, including electrocardiography (ECG), electroencephalography (EEG), thoracoabdominal distension, nasal airflow, and VNS artifacts. Measurements during stimulation were compared with those at baseline, immediately preceding the VNS periods and individually for each patient. RESULT: During VNS, respiratory frequency increased and respiratory amplitude decreased with a variable effect on cardiac activity. The coupling between heart rate and respiratory rate was disturbed and RSA magnitude decreased significantly in 6 of 10 children during VNS. These changes in RSA magnitude varied from one child to another. The observed changes for respiratory and cardiac activity were concomitant with changes in RSA but were not correlated. CONCLUSION: Together with disorders of respiration, cardiac activity, and oxygen saturation (SaO(2)) described previously. VNS also modifies synchronization between cardiac and respiratory activity, resulting in poor optimization of oxygen delivery to tissues that can be regarded as an additive side effect, which should be considered in patients with already altered brain function. This interaction between the effects of VNS and potential autonomic nervous system (ANS) dysfunction already reported in epileptic patients should be considered to be potentially life-threatening. In addition, evaluation of changes in respiratory parameters can also provide reliable markers for further evaluation of the effectiveness of VNS. PMID- 19682027 TI - How accurate is ICD coding for epilepsy? AB - PURPOSE: Assess the validity of ICD-9-CM and ICD-10 epilepsy coding from an emergency visit (ER) and a hospital discharge abstract database (DAD). METHODS: Two separate sources of patient records were reviewed and validated. (1) Charts of patients admitted to our seizure monitoring unit over 2 years (n = 127, ICD-10 coded records) were reviewed. Sensitivity (Sn), specificity (Sp), and positive and negative predictive values (PPV and NPV) were calculated. (2) Random sample of charts for patients seen in the ER or admitted to hospital under any services, and whose charts were coded with epilepsy or an epilepsy-like condition, were reviewed. Two time-periods were selected to allow validation of both ICD-9-CM (n = 486) and ICD-10 coded (n = 454) records. Only PPV and NPV were calculated for these records. All charts were reviewed by two physicians to confirm the presence/absence of epilepsy and compare to administrative coding. RESULTS: Sample 1: Sn, Sp, PPV, and NPV of ICD-10 epilepsy coding from the seizure monitoring unit (SMU) chart review were 99%, 70%, 85%, and 97% respectively. Sample 2: The PPV and NPV for ICD-9-CM coding from the ER database were, respectively, 99% and 97% and from the DAD were 98% and 99%. The PPV and NPV for ICD-10 coding from the ER database were, respectively, 100% and 90% and from the DAD were 98% and 99%. The epilepsy subtypes grand mal status and partial epilepsy with complex partial seizures both had PPVs >75% (ICD-9-CM and ICD-10 data). DISCUSSION: Administrative emergency and hospital discharge data have high epilepsy coding validity overall in our health region. PMID- 19682029 TI - Ictal apnea linked to contralateral spread of temporal lobe seizures: Intracranial EEG recordings in refractory temporal lobe epilepsy. AB - PURPOSE: Respiratory mechanisms are implicated in sudden unexpected death in epilepsy (SUDEP). We previously demonstrated a high incidence of ictal hypoxemia in temporal lobe seizures. We now report on the temporal relationship between ictal apnea and seizure onset and spread in patients undergoing video-EEG (electroencephalography) telemetry (VET) with intracranial electrodes. METHODS: Ten patients with medically refractory temporal lobe epilepsy (TLE) undergoing VET were studied. Data from synchronously recorded digital pulse oximetry (SaO(2)), end-tidal CO(2) (ETCO(2)), nasal airflow, abdominal excursions, and electrocardiography were obtained. RESULTS: Sixty-one seizures were captured. SaO(2) in the ictal/postictal period was available for 52 seizures, apnea onset times for 27 seizures, and ETCO(2) for 16 seizures. Apneas occurred only when seizures spread to the contralateral temporal lobe. The mean delay to apnea onset was significantly shorter after contralateral seizure spread (2.87 s) than after seizure onset (58.4 s); p < 0.001. The mean SaO(2) nadir with partial seizures or partial seizures prior to secondary generalization was 89.4 +/- 8.6% (91.5, 69 100). Following generalized convulsions the mean oxygen saturation nadir was 75.8 +/- 10.6% (78.5, 58-90). ETCO(2) elevations occurred with each ictal desaturation below 85%. CONCLUSIONS: There is a close temporal relationship between spread of seizures to the contralateral hemisphere and the onset of seizure-associated apnea. Apnea onsets are more tightly linked to time of contralateral spread than to time of seizure onset. Patients with TLE in whom there is evidence of contralateral seizure spread may be at higher risk for ictal-related respiratory dysfunction than those in whom seizures remain unilateral. PMID- 19682030 TI - Hippocampal volume assessment in temporal lobe epilepsy: How good is automated segmentation? AB - PURPOSE: Quantitative measurement of hippocampal volume using structural magnetic resonance imaging (MRI) is a valuable tool for detection and lateralization of mesial temporal lobe epilepsy with hippocampal sclerosis (mTLE). We compare two automated hippocampal volume methodologies and manual hippocampal volumetry to determine which technique is most sensitive for the detection of hippocampal atrophy in mTLE. METHODS: We acquired a three-dimensional (3D) volumetric sequence in 10 patients with left-lateralized mTLE and 10 age-matched controls. Hippocampal volumes were measured manually, and using the software packages Freesurfer and FSL-FIRST. The sensitivities of the techniques were compared by determining the effect size for average volume reduction in patients with mTLE compared to controls. The volumes and spatial overlap of the automated and manual segmentations were also compared. RESULTS: Significant volume reduction in affected hippocampi in mTLE compared to controls was detected by manual hippocampal volume measurement (p < 0.01, effect size 33.2%), Freesurfer (p < 0.01, effect size 20.8%), and FSL-FIRST (p < 0.01, effect size 13.6%) after correction for brain volume. Freesurfer correlated reasonably (r = 0.74, p << 0.01) with this manual segmentation and FSL-FIRST relatively poorly (r = 0.47, p << 0.01). The spatial overlap between manual and automated segmentation was reduced in affected hippocampi, suggesting the accuracy of automated segmentation is reduced in pathologic brains. DISCUSSION: Expert manual hippocampal volumetry is more sensitive than both automated methods for the detection of hippocampal atrophy associated with mTLE. In our study Freesurfer was the most sensitive to hippocampal atrophy in mTLE and could be used if expert manual segmentation is not available. PMID- 19682031 TI - Mossy fiber sprouting interacts with sodium channel mutations to increase dentate gyrus excitability. AB - PURPOSE: Idiopathic epilepsy is caused by the complex interaction of genetic and environmental factors. The purpose of this study was to use computational approaches to explore the interaction between changes in sodium channel availability caused by mutations and mossy fiber sprouting. METHODS: We used a previously published biophysically realistic computer model of dentate gyrus neurons and networks. A sensitivity analysis probed the effects of typical mutation-like changes in either single- or multiple-gating parameters. Isolated neuron models were stimulated with current injections, and networks were stimulated with perforant path synaptic input. The gene-environment interaction was studied by including mossy fiber sprouting into these networks. RESULTS: Single neuron responses to current injections were complex, with increased sodium channel availability paradoxically reducing firing rates. In the absence of mossy fiber sprouting, control networks showed strong accommodation supporting the role of the dentate gyrus as a gate. Availability changes alone were not able to drive the networks into ictal states, even though they reduced the effectiveness of the dentate gyrus gate. Interestingly, the presence of electrophysiologic changes substantially increased the ability of mossy fiber sprouting to induce ictal activity. CONCLUSION: (1) Increased sodium channel availability does not necessarily lead to increased firing rates, (2) network excitability is most sensitive to changes in steady state activation of sodium channels, (3) mutation induced changes in availability reduce the effectiveness of the dentate gyrus gate, and (4) mutations interact strongly with structural network changes to allow ictal-like activity in the dentate gyrus. PMID- 19682032 TI - Ethosuximide-induced de novo systemic lupus erythematosus with anti-double-strand DNA antibodies: a case report with definite evidence. PMID- 19682033 TI - Comment on "Factors influencing clinical features of absence seizures". PMID- 19682034 TI - Comment on "Aphasic or amnesic status epilepticus detected on PET but not EEG". PMID- 19682035 TI - Is the risk of vigabatrin-attributed visual field loss lower in infancy than in later life? PMID- 19682036 TI - Vigabatrin therapy in infantile spasms: solving one problem and inducing another? PMID- 19682039 TI - Landau-Kleffner syndrome: 50 years after. PMID- 19682040 TI - The Landau-Kleffner syndrome. PMID- 19682041 TI - Encephalopathy with status epilepticus during slow sleep: "the Penelope syndrome". AB - ESES (encephalopathy with status epilepticus during sleep) is an epileptic encephalopathy with heterogeneous clinical manifestations (cognitive, motor, and behavioral disturbances in different associations, and various seizure types) related to a peculiar electroencephalography (EEG) pattern characterized by paroxysmal activity significantly activated during slow sleep-that is, a condition of continuous spikes and waves, or status epilepticus, during sleep. The pathophysiologic mechanisms underlying this condition are still incompletely understood; recent data suggest that the abnormal epileptic EEG activity occurring during sleep might cause the typical clinical symptoms by interfering with sleep-related physiologic functions, and possibly neuroplasticity processes mediating higher cortical functions such as learning and memory consolidation. As in the myth of Penelope, the wife of Odysseus, what is weaved during the day will be unraveled during the night. PMID- 19682042 TI - Atypical rolandic epilepsy. AB - Typical benign rolandic epilepsy (BRE) is a frequent and well-delineated epileptic syndrome in childhood. Mild cognitive and behavioral difficulties are increasingly recognized in the course of BRE and should not be considered as atypical features. Atypical features are recognized on electroclinical grounds. These features, particularly early age at onset and frequent spikes or spike-wave discharges, seem to be risk factors for neuropsychological deficits but also for an atypical evolution of BRE. Atypical evolutions of BRE are defined by the appearance of severe neuropsychological impairments and continuous spike-and waves during slow sleep (CSWSS). The clinical expressions of these situations correspond to the syndromes known as atypical benign focal epilepsy of childhood (ABFEC), status of BRE, Landau-Kleffner syndrome (LKS), and CSWSS syndrome, which may be part of a continuum related to BRE. PMID- 19682043 TI - Guidelines for EEG in encephalopathy related to ESES/CSWS in children. AB - Electrical status epilepticus during slow sleep (ESES) or continuous spikes and waves during slow sleep (CSWS) is a phenomenon characterized by strong activation of epileptiform activity in the electroencephalogram (EEG) during sleep. The literature contains several small series of patients and many case reports. Large prospective studies are lacking. Definitions of the syndromes and EEG criteria and methods vary, as does their classification. The fluctuating clinical course and EEG findings complicate the diagnostic process and evaluation of effect of therapy. Studies describing quantitative aspects of the epileptiform abnormalities in EEG are overrepresented in literature, whereas qualitative aspects are relatively undervalued. Guidelines for evaluation of the EEG in these syndromes, which focus on both aspects, are presented. PMID- 19682044 TI - CSWS-related autistic regression versus autistic regression without CSWS. AB - Continuous spike-waves during slow-wave sleep (CSWS) and Landau-Kleffner syndrome (LKS) are two clinical epileptic syndromes that are associated with the electroencephalography (EEG) pattern of electrical status epilepticus during slow wave sleep (ESES). Autistic regression occurs in approximately 30% of children with autism and is associated with an epileptiform EEG in approximately 20%. The behavioral phenotypes of CSWS, LKS, and autistic regression overlap. However, the differences in age of regression, degree and type of regression, and frequency of epilepsy and EEG abnormalities suggest that these are distinct phenotypes. CSWS with autistic regression is rare, as is autistic regression associated with ESES. The pathophysiology and as such the treatment implications for children with CSWS and autistic regression are distinct from those with autistic regression without CSWS. PMID- 19682045 TI - Specific language impairment versus Landau-Kleffner syndrome. AB - The occurrence of sleep electroencephalography (EEG) abnormalities in some children with specific language impairment (SLI), the various forms of language dysfunction patterns seen in children with benign childhood epilepsy with centrotemporal spikes (BECTS), and finally the acquired aphasia in Landau Kleffner syndrome (LKS) indicate a large spectrum of interactions between language and epilepsy. As such, the question is whether SLI and LKS should rather be considered along a continuum or as two entirely distinct entities. In addition, the rationale for using antiepileptic medications in rare forms of SLI is discussed. PMID- 19682046 TI - From rolandic epilepsy to continuous spike-and-waves during sleep and Landau Kleffner syndromes: insights into possible genetic factors. AB - Epilepsy is a frequent neurologic disease in childhood, characterized by recurrent seizures and sometimes with major effects on social, behavioral, and cognitive development. Childhood focal epilepsies particularly are age-related diseases mainly occurring during developmental critical periods. A complex interplay between brain development and maturation processes and susceptibility genes may contribute to the development of various childhood epileptic syndromes associated with language and cognitive deficits. Indeed, the Landau-Kleffner syndrome (LKS), the continuous spike-and-waves during sleep syndrome (CSWS), and the benign childhood epilepsy with centrotemporal spikes (BCECTS) or benign rolandic epilepsy, are different entities that are considered as part of a single continuous spectrum of disorders. Genetic predisposition with simple to complex modes of inheritance has long been suspected for this wide group of childhood focal epilepsies. Recent reports on the involvement of the SRPX2 and ELP4 genes with possible roles in cell motility, migration, and adhesion have provided first insights into the complex molecular bases of childhood focal epilepsies. PMID- 19682047 TI - Acquired cognitive dysfunction with focal sleep spiking activity. AB - The syndrome of continuous spike-waves during slow sleep (CSWS) is considered an epileptic encephalopathy in which the epileptiform abnormalities may contribute to progressive cognitive dysfunction. The characteristic electroencephalographic feature of the syndrome occurs during non-REM sleep, and takes the form of continuous bilateral and diffuse slow spike-waves that persist through all slow sleep stages. Using a case study design including clinical, neuropsychological, electroencephalographic, and positron emission tomography with 18F fluorodeoxyglucose (PET-FDG) investigations, we describe the clinical and electroencephalographic findings in two patients who presented with nonsymptomatic epilepsy with unilateral spike-waves during sleep. Both patients presented with a left unilateral motor neglect of the upper limb that was associated with unilateral CSWS activity over the right hemisphere, predominantly in the centrotemporal region. PET-FDG studies during the active phase of CSWS showed right centrotemporal hypermetabolism in both cases. After treatment, a regression of the CSWS activity and an improvement of the cerebral FDG pattern were paralleled by a remission of the motor neglect. These cases demonstrate that the electroencephalographic pattern of CSWS in nonsymptomatic epilepsies is not necessarily diffuse and bilateral, and that focal unilateral CSWS activity can be associated with focal neuropsychological deficits. These findings add further evidence that the spectrum of clinical conditions associated with the electroencephalographic pattern of CSWS can include different forms of acquired cognitive disturbances that may be focal in nature. PMID- 19682048 TI - Neurophysiology of CSWS-associated cognitive dysfunction. AB - The phenomenon of continuous spikes and waves during slow-wave sleep (CSWS) is associated with a number of epileptic syndromes, which share a behavioral phenotype characterized by deterioration of cognitive, behavioral, or sensorimotor functions. Available evidence seems to suggest that spike-wave activity is a result of a complex interaction between cortical and subcortical inhibitory networks and can "per se" produce a transient loss of underlying cortical functions. Syndromes like Landau-Kleffner syndrome, CSWS, and phenomena such as negative myoclonus could share in common--at least at the neurophysiological level--some similarities. Differences in behavioral phenotypes could be explained in term of maturational and genetic differences, as well as by the functional specificity of the involved areas. PMID- 19682050 TI - Some facts about sleep relevant for Landau-Kleffner syndrome. AB - Our understanding of the neural mechanisms of non-rapid eye movement sleep (NREM) is steadily increasing. Given the intriguing activation of paroxysmal activity during NREM sleep in patients with Landau-Kleffner syndrome (LKS), a thorough characterization of commonalities and differences between the neural correlates of LKS paroxysms and normal sleep oscillations might provide useful information on the neural underpinning of this disorder. Especially, given the suspected role of sleep in brain plasticity, this type of information is needed to assess the link between cognitive deterioration and electroencephalography (EEG) paroxysms during sleep. PMID- 19682051 TI - Insights into the pathophysiology of psychomotor regression in CSWS syndromes from FDG-PET and EEG-fMRI. AB - Epilepsy syndromes with continuous spikes-and-waves during slow sleep (CSWS) are age-related epileptic encephalopathies characterized by the development of various types of psychomotor regression in close temporal concordance with the appearance of the electroencephalography (EEG) pattern of CSWS. Functional cerebral imaging studies performed in children with CSWS have shown evidence for the existence of increase in metabolism or perfusion at the site of the epileptic focus, associated with decrease in metabolism or perfusion in distant and connected brain areas. Longitudinal [18F]-fluorodeoxyglucose-positron emission tomography (FDG-PET) studies and effective connectivity analyses have suggested the existence of a pathophysiologic link between increases and decreases in metabolism/perfusion that could be explained by the theory of remote inhibition. These findings highlight that the psychomotor regression observed in CSWS syndromes is not only related to the neurophysiologic impairment at the site of the epileptic foci but also to epilepsy-induced neurophysiologic changes in distant connected brain areas. PMID- 19682049 TI - The epileptic hypothesis: developmentally related arguments based on animal models. AB - The significant morbidity linked to epileptic encephalopathies of childhood has prompted the need to identify and dissect the factors and mechanisms that contribute to the resultant functional regression. Although experiments specifically assessing language in rodents are difficult to design, a number of studies have shed light on the conditions that contribute to the functional deterioration. In particular, interictal spikes and seizures, especially if prolonged or frequent, may cause acute or long-lasting effects on brain functioning and development, which may impair performance in a variety of behavioral tests. These effects are further modified by a number of genetic, biological, and epigenetic factors, including age, sex, and underlying pathology, which further diversify outcome. Of special importance is the developmental age when the epileptic disorder manifests, because it may dictate outcome but also may be a deciding factor in selecting appropriate therapies. PMID- 19682052 TI - Magnetoencephalography in Landau-Kleffner syndrome. AB - Magnetoencephalography (MEG) detects weak magnetic fields outside the head. Spikes generated on the perisylvian convexity are detected only by electroencephalography (EEG), whereas spikes with intrasylvian generators are selectively seen by MEG. Generators of MEG and EEG spikes are determined using equivalent current dipoles (ECDs) that represent local cortical activity in parallel pyramidal neurons. MEG localizes cortical spike generators with a 1-2-cm spatial accuracy, and with a millisecond time resolution. This allows tracking of neural activity over successive synaptic connections in the cortical network. Both EEG and MEG are necessary for comprehensive spatial and temporal description of perisylvian epileptic networks in the Landau-Kleffner syndrome (LKS). MEG studies suggest that in more than 80% of LKS patients, the bilateral epileptic discharges are generated in the auditory- and language-related perisylvian cortex. Approximately 20% of children with LKS children have a unilateral perisylvian pacemaker that triggers secondary bilateral synchrony of spikes. This 20% may regain considerable language skills after multiple subpial transections (MSTs) of the pacemaker area. Outcome data of a few surgery patients suggest an indispensable role of MEG when planning the most efficient therapy for patients with LKS. PMID- 19682053 TI - Landau-Kleffner syndrome and CSWS syndrome: treatment with intravenous immunoglobulins. AB - This study reports results of therapy with immunoglobulin in children with Landau Kleffner syndrome (LKS) or the syndrome of continuous spikes and waves during sleep (CSWS syndrome). In a prospective study, children diagnosed between 2002 and 2006 with either LKS or CSWS syndrome were treated soon after diagnosis with intravenous courses of immunoglobulin (IVIg). We compared the results with those reported in the literature and with data from a retrospective survey of our earlier patients. Six children (two girls), aged 4-9 years, were included. Three had LKS, and three had CSWS syndrome. One child-with typical LKS-had been treated with prednisone before (without response). No patient had seizures during IVIg treatment and follow-up. Their electroencephalography (EEG) findings did not improve. Neuropsychological improvement occurred in one child with CSWS syndrome. Three children did not show any beneficial effect; they were subsequently treated with steroids, one with a clearly positive result. We conclude that successful treatment of LKS and CSWS syndrome with IVIg occurs occasionally. However, the improvement cannot always be clearly attributed to this. It might also reflect the natural course of the disease. Although the temporal relation between IVIg treatment and clinical improvement cannot be denied in individual patients, its real value remains to be determined. PMID- 19682054 TI - Rational treatment options with AEDs and ketogenic diet in Landau-Kleffner syndrome: still waiting after all these years. AB - Antiepileptic drugs (AEDs) remain a first treatment approach in Landau-Kleffner syndrome (LKS) and related syndromes. In the current literature, only class IV evidence is available. Inclusion criteria and outcome parameters are ill-defined. Most commonly, valproate, ethosuximide, and/or benzodiazepines are used. More recent case series show that sulthiame and especially levetiracetam can be considered as effective drugs. Smaller studies also point to the ketogenic diet as a valuable treatment option in LKS. PMID- 19682055 TI - The surgical treatment of Landau-Kleffner syndrome. AB - The medical management of Landau-Kleffner syndrome is usually effective for seizure control and eventual seizure remission. However, the response for language and behavior is often poor. Surgery, in the form of multiple subpial transections (MSTs) to include Wernicke's area has been suggested as a way forward if electrophysiologic lateralization can be demonstrated. Surgical series in the literature are few and outcome from surgery variable. PMID- 19682056 TI - Corticosteroids as treatment of epileptic syndromes with continuous spike-waves during slow-wave sleep. AB - To assess the efficacy and tolerability of steroids in epileptic syndromes with continuous spike-waves during slow-wave sleep (CSWS), charts of 44 children (25 boys) who received corticosteroids for cognitive and/or behavioral deterioration associated with CSWS were retrospectively reviewed. Awake and sleep electroencephalography (EEG) records, clinical and neuropsychological assessments were available before, during, and after corticosteroid therapy. Evaluation focused on effects on EEG, behavior, and cognition. All but two patients received hydrocortisone (initial dose of 5 mg/kg/day). The treatment was slowly tapered with a total duration of 21 months. There were 18 symptomatic and 26 cryptogenic cases. Mean age was 7 years and mean intelligence quotient/developmental quotient (IQ/DQ) was 65. Mean CSWS duration before corticosteroid treatment was 1.7 years. Twenty patients had tried more than two antiepileptic drugs (AEDs) before steroids. Positive response to steroids was found during the first 3 months of treatment in 34 of 44 patients (77.2%), with normalization of the EEG in 21 patients. Relapse occurred in 14 of them. Hence, 20 patients (45.4%) were long term responders after a single but prolonged trial of steroids, including all four cases of Landau-Kleffner syndrome. Positive response to steroids was highly significantly associated with higher IQ/DQ. Shorter CSWS duration, but not age, etiology, or previous AED trials, was associated with positive response to steroids. Early discontinuation of the treatment for side effects was encountered in seven patients. We conclude that corticosteroids are safe and efficient for treatment of epilepsy with CSWS. Poor responders are patients with very low IQ and long duration of CSWS. PMID- 19682057 TI - The assessment of auditory function in CSWS: lessons from long-term outcome. AB - In Landau-Kleffner syndrome (LKS), the prominent and often first symptom is auditory verbal agnosia, which may affect nonverbal sounds. It was early suggested that the subsequent decline of speech expression might result from defective auditory analysis of the patient's own speech. Indeed, despite normal hearing levels, the children behave as if they were deaf, and very rapidly speech expression deteriorates and leads to the receptive aphasia typical of LKS. The association of auditory agnosia more or less restricted to speech with severe language decay prompted numerous studies aimed at specifying the defect in auditory processing and its pathophysiology. Long-term follow-up studies have addressed the issue of the outcome of verbal auditory processing and the development of verbal working memory capacities following the deprivation of phonologic input during the critical period of language development. Based on a review of neurophysiologic and neuropsychological studies of auditory and phonologic disorders published these last 20 years, we discuss the association of verbal agnosia and speech production decay, and try to explain the phonologic working memory deficit in the late outcome of LKS within the Hickok and Poeppel dual-stream model of speech processing. PMID- 19682058 TI - Sign language in Landau-Kleffner syndrome. AB - This article reviews the history of sign language (SL) and the rationale for its use in children with profound auditory agnosia due to Landau-Kleffner syndrome (LKS), illustrated by studies of children and adults followed for many years and rare cases from the literature. The reasons that SL was successful and brought some children out of isolation while it could not be implemented in others are discussed. The nowadays earlier recognition and treatment of LKS and better awareness of the crucial need to maintain communication have certainly improved the outcome of affected children. Alternatives to oral language, even for less severe cases, are increasingly accepted. SL can be learned at different ages with a clear benefit, but the ambivalence of the patients and their families with the world and culture of the deaf may sometimes explain its refusal or limited acceptance. There are no data to support the fear that SL learning may delay or prevent oral language recovery in children with LKS. On the contrary, SL may even facilitate this recovery by stimulating functionally connected core language networks and by helping speech therapy and auditory training. PMID- 19682060 TI - Low-power laser to prevent oral mucositis in autologous hematopoietic stem cell transplantation. PMID- 19682059 TI - Massive intravascular haemolysis during Clostridium perfrigens sepsis of hepatic origin. PMID- 19682061 TI - Patient-specific recovery patterns over time measured by dependence in activities of daily living after stroke and post-stroke care: the South London Stroke Register (SLSR). AB - BACKGROUND AND PURPOSE: Data on patient-specific recovery after stroke are lacking and the effects of complex healthcare interventions on the course of recovery were not reported. To quantify the recovery pattern up to 1 year post stroke and assess effects of evidence-based treatments on the patient-specific course of recovery allowing its prediction. METHODS: A total of 355 patients after first-ever stroke from the population-based South London Stroke Register (source population >270,000) participated in a substudy between August 2002 and October 2004. At 1, 2, 3, 4, 6, 8, 12, 26, and 52 weeks post-stroke, Barthel Index (BI; ranging from 0 to 20) was documented. Multilevel growth models allowing predictions for patients with specific characteristics were calculated. RESULTS: Mean age was 70 years, 48% were male and 23% died within the first year. The age-, gender- and stroke subtype-adjusted BI curve sharply increased until week 8 to 24 depending on patient characteristics and subsequently plateaued. Multivariable analysis identified stroke unit care, appropriate secondary prevention and physiotherapy for those with disabilities as independent predictors of improved functional ability over time (P < 0.05). Patients receiving stroke unit care additionally gained 4 BI points within 6 months compared with their counterparts (P = 0.004). CONCLUSIONS: Functional outcome in the general population showed an increase followed by a plateau. Care parameters reflecting guideline treatment independently improved recovery illustrating the beneficial effects of evidence-based interventions on recovery in an unbiased population. PMID- 19682062 TI - A 12-month longitudinal study of calcium metabolism and bone turnover during valproate monotherapy. AB - BACKGROUND AND PURPOSE: Treatment with valproate (VPA) can cause changes in bone mineral metabolism, but the real influences and the underlying pathologic mechanisms are still unclear and under discussion. The aim of this study was to examine the changes on calcium metabolism and bone turnover in post-pubertal male patients with newly diagnosed idiopathic generalized epilepsy (IGE) before (baseline evaluation) and 12 months after VPA monotherapy (second evaluation). METHODS: Participants included 20 post-pubertal males with IGE, aged 16.5-22.1 years. Also 20 post-pubertal sex- and age-matched healthy controls were evaluated. Physical activity, calcium and vitamin D intake were determined. Laboratory samples were obtained to measure biochemical parameters of bone metabolism and bone turnover: serum calcium, phosphate, magnesium, 25 hydroxyvitamin D, 1,25-dihydroxyvitamin D, intact parathyroid hormone, total alkaline phosphatase, bone alkaline phosphatase (bone-ALP), osteocalcin (OC), carboxy-terminal propeptide of type I procollagen (PICP) and carboxy-terminal telopeptide of type I collagen (ICTP). RESULTS: At baseline evaluation, there were no significant differences between controls and patients parameters. At second evaluation, patients showed both markers of bone formation and resorption significantly higher than baseline values (bone-ALP: 51.2 +/- 9.9 vs. 57.3 +/- 9.3 U/l, P < 0.01; OC: 8.1 +/- 1.1 vs. 10.4 +/- 1.5 microg/l, P < 0.01; PICP: 138.7 +/- 16.4 vs. 152.6 +/- 17.1 microg/l, P < 0.01; ICTP: 3.8 +/- 0.8 vs. 5.9 +/- 0.6 microg/l, P < 0.01). CONCLUSIONS: Valproate monotherapy in epileptic post pubertal males causes a significant increase of bone turnover. PMID- 19682063 TI - Hyperleukocytotic secondary acute myeloid leukemia (AML) with sole monosomy 7 as sequela of Philadelphia-chromosome positive chronic myeloid leukemia (CML). PMID- 19682064 TI - Characterisation of hairy cell leukaemia by tiling resolution array-based comparative genome hybridisation: a series of 13 cases and review of the literature. AB - Little is known about the cytogenetic features and molecular mechanisms behind hairy cell leukaemia (HCL), despite the advances in diagnosis and treatment. Therefore, we used high-resolution genome-wide array-based comparative genomic hybridisation (array-CGH) and multiplex ligation-dependent probe amplification (MLPA) to characterise copy number alterations (CNAs) in DNA from 13 cases of HCL. We also summarise CNAs and cytogenetic features in 109 HCL cases comprising our 13 cases and 96 cases from the literature. Genomic array-CGH revealed imbalances in two out of 13 cases in addition to previously described copy number variants (CNVs) found in healthy individuals. In one case, a 700 kb deletion of 20q11.22 was detected encompassing ten characterised genes, among them the TP53INP2, DNCL2A and ITCH genes. In the second case, trisomy 5, and a deletion of 5p15.2 encompassing a non-characterised gene AY328033 was found. Altogether only 20/81 (25%) of all cases studied by CGH or gene dose array revealed CNAs. The most common recurrent deletions and breakpoints were 14q22-32 (33%), 6q25 (16%), 2p12 (10%), 22q11 (10%), 17p11-13 (10%), 7q32-36 (9%), 18q11-13 (7%), 1q32-44 (6%), 8p22-23 (6%) and 7q11 (6%). Trisomy 5 occurred in 15%. In addition, several other recurrent breakpoints were identified. Although a number of genomic imbalances were identified in the HCL samples, the genome appeared remarkably stable. PMID- 19682065 TI - Osteopenia and vitamin D deficiency in children with sickle cell disease. AB - OBJECTIVES: To assess the prevalence in children with sickle cell disease of low bone mineral density (BMD), a feature found in up to 82% of adults but not well known in children. METHODS: In 53 children (45 SS, 4 SC, 4 Sbeta-thalassemia) with a mean age of 12.8 +/- 2.4 years, we assessed height; weight; sexual maturation; number of hospitalizations, painful crises, and transfusions in the last 3 years; calcium intake; steady-state hemoglobin and leukocyte count; calcaemia, phosphataemia, and calciuria/creatinuria; serum 25-(OH)D and PTH concentrations; and osteocalcin, urinary deoxypyridinoline, and the C-terminal component of pro-collagen type I. BMD was assessed using dual X-ray absorptiometry. RESULTS: Mean lumbar spine Z-score was -1.1 +/- 1.3 (-3.9 to +1.8). The Z score was significantly lower in girls than in boys in the prepubertal subgroup (-1.74 +/- 0.27 vs. -0.53 +/- 0.31) (P = 0.0169), but not in the pubertal group (-1.15 +/- 0.41 vs. -1.33 +/- 0.70). BMD was not associated with any of the disease-severity markers in girls but was unexpectedly associated with fewer vaso-occlusive crises and hospitalizations in boys. BMD did not correlate with hemoglobin or leukocyte counts. Vitamin D deficiency [25-(OH)D < 12 ng/mL] was found in 76% of patients and secondary hyperparathyroidism (PTH > 46 pg/mL) in 38%. BMD was not related to calcium intake, vitamin D status, osteocalcin, or bone resorption markers. CONCLUSION: A slight BMD decrease was found in SCD children, starting before puberty and being more marked in females. The decrease was unrelated to disease severity, vitamin D deficiency, or bone hyperresorption, suggesting abnormal bone formation as the underlying mechanism. PMID- 19682066 TI - Intra-atrial thrombosis complicating myocardial relapse of leukaemia. PMID- 19682067 TI - Life at the front of an expanding population. AB - Environmental changes have caused episodes of habitat expansions in the evolutionary history of many species. These range changes affect the dynamics of biological evolution in multiple ways. Recent microbial experiments as well as simulations suggest that enhanced genetic drift at the frontier of a two dimensional range expansion can cause genetic sectoring patterns with fractal domain boundaries. Here, we propose and analyze a simple model of asexual biological evolution at expanding frontiers that explains these neutral patterns and predicts the effect of natural selection. We find that beneficial mutations give rise to sectors with an opening angle that depends sensitively on the selective advantage of the mutants. Deleterious mutations, on the other hand, are not able to establish a sector permanently. They can, however, temporarily "surf" on the population front, and thereby reach unusually high frequencies. As a consequence, expanding frontiers are loaded with a high fraction of mutants at mutation-selection balance. Numerically, we also determine the condition at which the wild type is lost in favor of deleterious mutants (genetic meltdown) at a growing front. Our prediction for this error threshold differs qualitatively from existing well-mixed theories, and sets tight constraints on sustainable mutation rates for populations that undergo frequent range expansions. PMID- 19682068 TI - Molecular characterization of Arabidopsis thaliana PUF proteins--binding specificity and target candidates. AB - PUF proteins regulate both stability and translation through sequence-specific binding to the 3' UTR of target mRNA transcripts. Binding is mediated by a conserved PUF domain, which contains eight repeats of approximately 36 amino acids each. Found in all eukaryotes, they have been related to several developmental processes. Analysis of the 25 Arabidopsis Pumilio (APUM) proteins presenting PUF repeats reveals that 12 (APUM-1 to APUM-12) have a PUF domain with 50-75% similarity to the Drosophila PUF domain. Through three-hybrid assays, we show that APUM-1 to APUM-6 can bind specifically to the Nanos response element sequence recognized by Drosophila Pumilio. Using an Arabidopsis RNA library in a three-hybrid screening, we were able to identify an APUM-binding consensus sequence. Computational analysis allowed us to identify the APUM-binding element within the 3' UTR in many Arabidopsis transcripts, even in important mRNAs related to shoot stem cell maintenance. We demonstrate that APUM-1 to APUM-6 are able to bind specifically to APUM-binding elements in the 3' UTR of WUSCHEL, CLAVATA-1, PINHEAD/ZWILLE and FASCIATA-2 transcripts. The results obtained in the present study indicate that the APUM proteins may act as regulators in Arabidopsis through an evolutionarily conserved mechanism, which may open up a new approach for investigating mRNA regulation in plants. PMID- 19682069 TI - The miRNA-192/194 cluster regulates the Period gene family and the circadian clock. AB - Several biological functions in mammals are regulated in a circadian fashion. The molecular mechanisms orchestrating these circadian rhythms have been unravelled. The biological clock, with its core transcriptional unit Bmal1/CLOCK, is composed of several self-sustaining feedback loops. In this study, we describe another mechanism impinging on the core components of the circadian clock. Using a forward genetic screen, we identified the miR-192/194 cluster as a potent inhibitor of the entire Period gene family. In accordance, the exogenous expression of miR-192/194 leads to an altered circadian rhythm. Thus, our results have uncovered a new mechanism for the control of the circadian clock at the post transcriptional level. PMID- 19682070 TI - Ca2+/calmodulin-dependent kinase II signalling cascade mediates P2X7 receptor dependent inhibition of neuritogenesis in neuroblastoma cells. AB - ATP, via purinergic P2X receptors, acts as a neurotransmitter and modulator in both the central and peripheral nervous systems, and is also involved in many biological processes, including cell proliferation, differentiation and apoptosis. Previously, we have reported that P2X7 receptor inhibition promotes axonal growth and branching in cultured hippocampal neurons. In this article, we demonstrate that the P2X7 receptor negatively regulates neurite formation in mouse Neuro-2a neuroblastoma cells through a Ca2+/calmodulin-dependent kinase II related mechanism. Using both molecular and immunocytochemical techniques, we characterized the presence of endogenous P2X1, P2X3, P2X4 and P2X7 subunits in these cells. Of these, the P2X7 receptor was the only functional receptor, as its activation induced intracellular calcium increments similar to those observed in primary neuronal cultures, exhibiting pharmacological properties characteristic of homomeric P2X7 receptors. Patch-clamp experiments were also conducted to fully demonstrate that ionotropic P2X7 receptors mediate nonselective cation currents in this cell line. Pharmacological inhibition of the P2X7 receptor and its knockdown by small hairpin RNA interference resulted in increased neuritogenesis in cells cultured in low serum-containing medium, whereas P2X7 overexpression significantly reduced the formation of neurites. Interestingly, P2X7 receptor inhibition also modified the phosphorylation state of focal adhesion kinase, Akt and glycogen synthase kinase 3, protein kinases that participate in the Ca2+/calmodulin-dependent kinase II signalling cascade and that have been related to neuronal differentiation and axonal growth. Taken together, our results provide the first mechanistic insight into P2X7 receptor-triggered signalling pathways that regulate neurite formation in neuroblastoma cells. PMID- 19682071 TI - The yeast ubiquitin ligase Rsp5 downregulates the alpha subunit of nascent polypeptide-associated complex Egd2 under stress conditions. AB - The ubiquitination and subsequent degradation of stress-induced abnormal proteins are indispensable to cell survival. We previously showed that a yeast (Saccharomyces cerevisiae) mutant carrying a single amino acid change, Ala401Glu, in RSP5, which encodes an essential E3 ubiquitin ligase, is hypersensitive to various stresses. To identify the protein substrates of Rsp5, we performed a comparative proteome analysis of the wild-type and rsp5 mutant strains under stress conditions. The results we obtained indicate that several proteins, including the a-subunit of nascent polypeptide-associated complex (aNAC, Egd2) accumulated in the rsp5 mutant. To investigate whether or not Rsp5 ubiquitinates these proteins in a stress-dependent manner, cell extracts were analyzed by immunoprecipitation followed by western blotting after exposure to temperature upshift. Interestingly, Egd2 was ubiquitinated in the wild-type cells but not in the rsp5 mutant cells under these stress conditions. We also detected in vitro ubiquitination of Egd2 by Rsp5 at elevated temperature. Moreover, Egd2 was ubiquitinated in the egd1 and not4 deletion mutants lacking bNAC and the RING type ubiquitin ligase Not4, respectively, indicating that ubiquitination of Egd2 is independent of Egd1 and Not4. We also showed that, under stress conditions, Egd2 was mainly degraded via the proteasome pathway. These results strongly suggest that Rsp5 is involved in selective ubiquitination and degradation of stress-induced unstable proteins, such as Egd2. PMID- 19682072 TI - Increased bacterial load in shrimp hemolymph in the absence of prophenoloxidase. AB - Invertebrates rely on their innate immune responses to protect themselves from pathogens, one of which is melanization of bacteria mediated by the activation of phenoloxidase (PO). Furthermore, invertebrate hemolymph, even that of healthy individuals, has been shown to contain bacterial species. The mechanisms that prevent these bacteria from proliferating and becoming deleterious to the host are, however, poorly understood. Here, we show that knocking down the activity of the inactive precursor of PO [prophenoloxidase (proPO)] by RNA interference resulted in a significant increase in the bacterial load of kuruma shrimp, Marsupenaeus japonicus, even in the absence of a bacterial or viral challenge. Silencing of proPO also led to a sharp increase in shrimp mortality. In addition, the hemolymph of proPO-depleted shrimp had significantly lower hemocyte counts and PO activity than control samples. Microarray analysis after proPO silencing also showed a decrease in the expression of a few antimicrobial peptides, but no effect on the expression of the genes involved in the clotting system. Treatment with antibiotics prior to and after proPO dsRNA injection, to counteract the loss of proPO, resulted in a significant increase in shrimp survival. Our results therefore show that the absence of proPO renders the shrimp incapable of controlling bacteria present in the hemolymph, and that proPO is therefore essential for its survival. PMID- 19682073 TI - Collagen I regulates matrix metalloproteinase-2 activation in osteosarcoma cells independent of S100A4. AB - This work investigates the effect of cell-collagen I interactions on the synthesis and activation of MMP-2, as well as synthesis of MT1-MMP and TIMP-1, by using an in vitro model with 3D fibrillar and 2D monomeric collagen. In order to reveal whether the metastasis-associated protein S100A4 can influence the cell's response to the two forms of collagen, osteosarcoma cell lines with high and low endogenous levels of S100A4 were used. Attachment of osteosarcoma cells to 3D fibrillar and 2D monomeric collagen resulted in opposite effects on MMP-2 activation. Attachment to 3D fibrillar collagen decreased activation of proMMP-2, with a corresponding reduction in MT1-MMP. By contrast, attachment to monomeric collagen increased the amount of fully active MMP-2. This was caused by a reduction in TIMP-1 levels when cells were attached to monomeric 2D collagen. The effect of collagen on proMMP-2 activation was independent of endogenous S100A4 levels, whereas synthesis of TIMP-1 was dependent on S100A4. When cells were attached to monomeric collagen, cells with a high level of S100A4 showed a greater reduction in the synthesis of TIMP-1 than did those with a low level of S100A4. Taken together, this study shows that synthesis and activation of MMP-2 is affected by interactions between osteosarcoma cells and collagen I in both fibrillar and monomeric form. PMID- 19682074 TI - A single intersubunit salt bridge affects oligomerization and catalytic activity in a bacterial quinone reductase. AB - YhdA, a thermostable NADPH:FMN oxidoreductase from Bacillus subtilis, reduces quinones via a ping-pong bi-bi mechanism with a pronounced preference for NADPH. The enzyme occurs as a stable tetramer in solution. The two extended dimer surfaces are packed against each other by a 90 rotation of one dimer with respect to the other. This assembly is stabilized by the formation of four salt bridges between K109 and D137 of the neighbouring protomers. To investigate the importance of the ion pair contacts, the K109L and D137L single replacement variants, as well as the K109L/D137L and K109D/D137K double replacement variants, were generated, expressed, purified, crystallized and biochemically characterized. The K109L and D137L variants form dimers instead of tetramers, whereas the K109L/D137L and K109D/D137K variants appear to exist in a dimer tetramer equilibrium in solution. The crystal structures of the K109L and D137L variants confirm the dimeric state, with the K109L/D137L and K109D/D137K variants adopting a tetrameric assembly. Interestingly, all protein variants show a drastically reduced quinone reductase activity in steady-state kinetics. Detailed analysis of the two half reactions revealed that the oxidative half reaction is not affected, whereas reduction of the bound FMN cofactor by NADPH is virtually abolished. Inspection of the crystal structures indicates that the side chain of K109 plays a dual role by forming a salt bridge to D137, as well as stabilizing a glycine-rich loop in the vicinity of the FMN cofactor. In all protein variants, this glycine-rich loop exhibits a much higher mobility, compared to the wild type. This appears to be incompatible with NADPH binding and thus leads to abrogation of flavin reduction. PMID- 19682075 TI - The carbohydrate-binding module family 20--diversity, structure, and function. AB - Starch-active enzymes often possess starch-binding domains (SBDs) mediating attachment to starch granules and other high molecular weight substrates. SBDs are divided into nine carbohydrate-binding module (CBM) families, and CBM20 is the earliest-assigned and best characterized family. High diversity characterizes CBM20s, which occur in starch-active glycoside hydrolase families 13, 14, 15, and 77, and enzymes involved in starch or glycogen metabolism, exemplified by the starch-phosphorylating enzyme glucan, water dikinase 3 from Arabidopsis thaliana and the mammalian glycogen phosphatases, laforins. The clear evolutionary relatedness of CBM20s to CBM21s, CBM48s and CBM53s suggests a common clan hosting most of the known SBDs. This review surveys the diversity within the CBM20 family, and makes an evolutionary comparison with CBM21s, CBM48s and CBM53s, discussing intrafamily and interfamily relationships. Data on binding to and enzymatic activity towards soluble ligands and starch granules are summarized for wild-type, mutant and chimeric fusion proteins involving CBM20s. Noticeably, whereas CBM20s in amylolytic enzymes confer moderate binding affinities, with dissociation constants in the low micromolar range for the starch mimic beta cyclodextrin, recent findings indicate that CBM20s in regulatory enzymes have weaker, low millimolar affinities, presumably facilitating dynamic regulation. Structures of CBM20s, including the first example of a full-length glucoamylase featuring both the catalytic domain and the SBD, are summarized, and distinct architectural and functional features of the two SBDs and roles of pivotal amino acids in binding are described. Finally, some applications of SBDs as affinity or immobilization tags and, recently, in biofuel and in planta bioengineering are presented. PMID- 19682076 TI - Structural and genetic characterization of Escherichia coli O99 antigen. AB - O-antigen is part of the lipopolysaccharide present in the outer membrane of Gram negative bacteria, and contributes the major antigenic variability to the cell surface. Screening for the Escherichia coli O-serogroup is the conventional method for identifying E. coli clones. In this study, we investigated the structural characteristics of the E. coli O99 O-antigen and the organization of the genes involved in its synthesis. On the basis of sugar and methylation analysis and nuclear magnetic resonance spectroscopy data, we established the structure of the branched hexasaccharide repeat unit of the O-polysaccharide. This unit consists of four d-rhamnose (d-Rha) moieties in the backbone and two d glucose (d-Glc) moieties in the side chain, as shown below: [carbohydrate structure: see text]. The O-antigen gene cluster of E. coli O99, which was located between galF and gnd, was found to contain putative genes for the synthesis of d-Rha, genes encoding sugar transferases, and ATP-binding cassette (ABC) transporter genes (wzm and wzt). Our findings indicate that in E. coli O99, the synthesis and translocation of the O-antigen occurs by an ABC transporter dependent process. PMID- 19682077 TI - Helicobacter equorum: prevalence and significance for horses and humans. AB - Helicobacter equorum colonizes the caecum, colon and rectum of horses. The agent is highly prevalent in <6-month-old foals. In adult horses, the prevalence of H. equorum seems to be rather low, but these animals may harbour low, subdetectable numbers of this microorganism in their intestines. So far, no association could be made between the presence of H. equorum and clinical disease or intestinal lesions in adult horses. Further research is necessary to elucidate the pathogenic potential of this bacterial species towards young foals. Helicobacter equorum DNA was not detected in human faeces, indicating that this microorganism does not commonly spread from horses towards humans. PMID- 19682079 TI - Lasting reduction of postsurgical hyperalgesia after single injection of botulinum toxin type A in rat. AB - A single injection of low doses of botulinum toxin type A (3.5 U/kg) completely abolished secondary mechanical hyperalgesia throughout its duration in a model of post surgical pain after gastrocnemius incision in rat. PMID- 19682078 TI - Identification of Chlamydia trachomatis CT621, a protein delivered through the type III secretion system to the host cell cytoplasm and nucleus. AB - Chlamydiae are obligate intracellular bacteria, developing inside host cells within chlamydial inclusions. From these inclusions, the chlamydiae secrete proteins into the host cell cytoplasm. A pathway through which secreted proteins can be delivered is the type III secretion system (T3SS). The T3SS is common to several gram-negative bacteria and the secreted proteins serve a variety of functions often related to the modulation of host signalling. To identify new potentially secreted proteins, the cytoplasm was extracted from Chlamydia trachomatis L2-infected HeLa cells, and two-dimensional polyacrylamide gel electrophoresis profiles of [(35)S]-labelled chlamydial proteins from this extract were compared with profiles of chlamydial proteins from the lysate of infected cells. In this way, CT621 was identified. CT621 is a member of a family of proteins containing a domain of unknown function DUF582 that is only found within the genus Chlamydia. Immunofluorescence microscopy and immunoblotting demonstrated that CT621 is secreted late in the chlamydial developmental cycle and that it is the first chlamydial protein found to be localized within both the host cell cytoplasm and the nucleus. To determine whether CT621 is secreted through the T3SS, an inhibitor of this apparatus was added to the infection medium, resulting in retention of the protein inside the chlamydiae. Hence, the so far uncharacterized CT621 is a new type III secretion effector protein. PMID- 19682080 TI - Combined therapy in the treatment of dyslipidemia. AB - This systematic review analyses the efficacy, tolerability and safety of combinations of different medicines used to treat dyslipidemias in clinical practice. A PubMed search up to January 2009, was conducted to identify relevant studies. Criteria used to identify studies included (1) English language, (2) published studies with original data or meta-analyses in peer-reviewed journals. Although statin treatment is a mainstay of dyslipidemia management today, complementary effects of other lipid-lowering and/or HDL-cholesterol-raising therapies might substantially increase the clinical benefits not only in the small minority of patients with severe dyslipidemias but in others as well. These therapies include combinations with bile acid sequestrants (cholestyramine, colestipol, colesevelam), ezetimibe, niacin, plant sterols, fibrates (fenofibrate, bezafibrate, gemfibrozil), and prescription omega-3 fatty acids. Therapeutic approaches which incorporate the use of multiple drugs combinations for dyslipidemia treatment should be more widely adopted since combination therapy might offer a means to increase the number of patients able to meet their lipoprotein goals according to the guidelines. However, it has to be stated that for most of these combination therapies data on cardiovascular outcomes are still lacking. PMID- 19682081 TI - Effect of chronic pre-treatment with angiotensin converting enzyme inhibition on skeletal muscle mitochondrial recovery after ischemia/reperfusion. AB - Impaired skeletal muscle energetic participates in peripheral arterial disease (PAD) patient's morbidity and mortality. Angiotensin converting enzyme inhibition (ACEi), cornerstone for pharmacologic risk factor management in PAD patients, might also be interesting by protecting skeletal muscle energetic. We therefore determined whether chronic ACEi might reduce ischemia-induced mitochondrial respiratory chain dysfunction in the frequent setting of hindlimb ischemia reperfusion. Ischemic legs of rats submitted to 5 h ischemia induced by a rubber band tourniquet applied on the root of the hindlimb followed by reperfusion without (IR, n = 11) or after ACEi (n = 14; captopril 40 mg/kg per day during 28 days before surgery) were studied and compared to that of sham-operated animals (n = 11). The effect of ACEi on the non-ischemic contralateral leg was also determined in the ACEi group. Maximal oxidative capacities (V(max)) and complexes I, II and IV activities of the mitochondrial respiratory chain of the gastrocnemius muscle were determined using glutamate-malate, succinate and TMPD ascorbate substrates. Arterial blood pressure was significantly decreased after ACEi (124 +/- 2.8 vs. 108 +/- 4.19 mmHg; P = 0.01). Ischemia-reperfusion reduced V(max) (4.4 +/- 0.4 vs. 8.7 +/- 0.5 micromol O2/min/g dry weight, -49%, P < 0.001), affecting mitochondrial complexes I, II and IV activities. ACEi failed to modulate ischemia-induced dysfunction (V(max) 5.1 +/- 0.7 micromol O2/min/g dry weight) or the non-ischemic contralateral muscle respiratory rate. Ischemia reperfusion significantly impaired the mitochondrial respiratory chain I, II and IV complexes of skeletal muscle. Pharmacologic pre-treatment with ACEi did not prevent or increase such alterations. Further studies might be useful to improve the pharmacologic conditioning of PAD patients needing arterial revascularization. PMID- 19682082 TI - Protective effect of p-methoxyl-diphenyl diselenide in lethal acute liver failure induced by lipopolysaccharide and D-galactosamine in mice. AB - Organoselenium compounds display important antioxidant activity and many biological activities interesting from pharmacological point of view. The aim of this study was to evaluate the hepatoprotective effect of p-methoxyl-diphenyl diselenide, a disubstituted diaryl diselenide, on acute liver injury induced by D galactosamine (D-GalN) and lipopolysaccharide (LPS) in mice. The animals received p-methoxyl-diphenyl diselenide (10, 50 and 100 mg/kg; per oral, p.o.) and 1 h after d-GalN (500 mg/kg) and LPS (50 microg/kg) were administered by intraperitoneal route (i.p.). Twenty-four hours after LPS/d-GalN exposure the animals were euthanized to the biochemical and histological analysis. Pretreatment with p-methoxyl-diphenyl diselenide (50 and 100 mg/kg; p.o.) protected against the increase in aspartate aminotransferase (AST) activity induced by LPS/d-GalN exposure in mice. p-Methoxyl-diphenyl diselenide at the doses of 50 and 100 mg/kg protected against the increase in alanine aminotransferase (ALT) activity induced by LPS/D-GalN exposure. In this study, no alteration in ascorbic acid levels was observed in livers of mice exposed to LPS/D-GalN. Glutathione-S-transferase (GST) activity was stimulated by LPS/D-GalN exposure and p-methoxyl-diphenyl diselenide, at all doses, protected against this alteration. p-Methoxyl-diphenyl diselenide was effective in ameliorating inhibition of catalase activity induced by LPS/d-GalN exposure. Histological data showed that sections of livers from LPS/D-GalN-treated mice presented massive hemorrhage, inflammatory cells and necrosis. p-Methoxyl-diphenyl diselenide significantly attenuated LPS/D-GalN-induced hepatic histopathological alterations. Based on the results, we suggest the hepatoprotective effect of p methoxyl-diphenyl diselenide on acute liver injury induced by LPS/d-GalN exposure in mice. PMID- 19682083 TI - Haplotype structure and allele frequencies of CYP2B6 in Spaniards and Central Americans. AB - This study was aimed to investigate the potential differences in allele frequencies of the CYP2B6 gene between Spaniards and Central Americans. Three single nucleotide polymorphisms of the CYP2B6 gene 516 G>T, 785 A>G and 1459 C>T were assayed by a polymerase chain reaction in 180 Spaniards and 182 Central Americans. The allele frequencies for CYP2B6*1, CYP2B6*4, CYP2B6*5, CYP2B6*6, CYP2B6*9 in Spaniards and Central Americans were 0.593 and 0.642, 0.062 and 0.073, 0.113 and 0.030, 0.215 and 0.230, 0.014 and 0.023, respectively. CYP2B6*5 was less prevalent among Central Americans than in Spaniards (P < 0.001). In comparison to other previously studied populations, the CYP2B6*5 allele frequency among Spaniards was similar to other Caucasian or African groups, and higher than that in Asian populations. The CYP2B6*5 allele frequency in Central Americans was lower than that in Africans or Caucasian groups and higher than in Asians. The results indicate the presence of ethnic differences in CYP2B6 genetic variants between Spaniards and Central Americans, and support the need for further investigations to explore whether these differences significantly alter the efficacy or toxicity of CYP2B6 substrate drugs. PMID- 19682084 TI - Statins, fibrates, nicotinic acid, cholesterol absorption inhibitors, anion exchange resins, omega-3 fatty acids: which drugs for which patients? AB - Classes of lipid lowering drugs differ strongly with respect to the types of lipids or lipoproteins they predominantly affect. Statins inhibit the de-novo synthesis of cholesterol. Consequently, the liver produces less VLDL, and the serum concentration primarily of LDL cholesterol (but, to a lesser extent, also of triglycerides) is lowered. Further, statins somewhat increase HDL cholesterol. There is abundant evidence that statins lower the rate of cardiovascular events. Cardiovascular risk reduction is the better, the lower the LDL cholesterol values achieved with statin therapy are. Some evidence is available that anion exchange resins which also decrease LDL cholesterol decrease vascular risk, too. This is not the case for the ezetimibe, which strongly lowers LDL cholesterol: its potential to decrease vascular risk remains to be proven. In contrast evidence for cardiovascular risk reduction through the mainly triglyceride lowering fibrates as well as for niacin is available. Niacin is the most potent HDL increasing drug currently available and besides increasing HDL cholesterol efficaciously lowers triglycerides and LDL cholesterol. Large ongoing trials address the decisive question whether treatment with fibrates and niacin provides additional cardiovascular risk reduction when given in addition to statin treatment. PMID- 19682085 TI - Association between inosine triphosphate pyrophosphohydrolase deficiency and azathioprine-related adverse drug reactions in the Chinese kidney transplant recipients. AB - Azathioprine (AZA) is a thiopurine prodrug commonly used in patients with kidney transplantation. The aim of this study is to explore in patients with kidney transplantation whether AZA-related side effects can be explained by the inosine triphophate pyrophosphatase (ITPA) or thiopurine S-methyltransferase (TPMT) polymorphisms using both pheno-and genotyping. Erythrocyte ITPA and TPMT activity of 155 patients with kidney transplantation and AZA therapy was determined by HPLC. The frequencies of ITPA and TPMT polymorphisms were detected. Among 155 patients, three cases with zero activity were homozygote for 94C>A. The allele frequency of the 94C>A polymorphism was 0.12. Allele for the IVS2+21A>C mutation in the patients of this study was not found. Thirty-five cases had stopped azathioprine medication or were on reduced dose due to AZA-related side effects, including hematotoxicity (n = 12), hepatotoxicity (n = 18), gastrointestinal toxicity (n = 5, one patient developed hepatotoxicity simultaneously) and flu like symptoms (n = 1). No statistical significant associations between ITPA 94C>A phenotype or genotype and AZA-related hematotoxicity or hepatotoxicity could be detected. However, five patients who developed gastrointestinal disturbance, two patients were homozygote for 94C>A and other three patients had 94C>A heterozygous allele. The patient who experienced flu-like symptoms were the remaining homozygote for 94C>A. This study demonstrates that ITPA activity reduced in patients with 94C>A mutation (P < 0.01). Patients with ITPA 94C>A homozygous allele are at high risk to develop AZA-related gastrointestinal toxicity and flu-like symptoms (P < 0.01). TPMT wild-type/ITPA variant (homozygote) is closely related to the AZA-induced side effects (P < 0.01). PMID- 19682086 TI - Vasorelaxant effects of the monoterpenic phenol isomers, carvacrol and thymol, on rat isolated aorta. AB - Various essential oils are rich in carvacrol, a monoterpenic phenol isomeric with thymol. This study was undertaken to assess the vasorelaxant effects of thymol and carvacrol in rat isolated aorta and the putative mechanisms underlying these effects. Thymol and carvacrol produced a concentration-dependent relaxation on the aortic ring preparations pre-contracted using KCl (IC(50) value of 64.40 +/- 4.41 and 78.80 +/- 11.91 microm, respectively) or using phenylephrine (PHE, 0.1 microm) (IC(50) value of 106.40 +/- 11.37 and 145.40 +/- 6.07 microm, respectively) and inhibited the concentration-response curves of aortic rings to PHE or KCl. In Ca(2+)-free medium with ethylene glycol-bis(2-aminoethylether) N,N,N',N'-tetraacetic acid (2 mm), thymol and carvacrol both at 1000 microm completely abolished the phasic component of PHE-induced endothelium-containing ring contractions. At 400 microm, thymol and carvacrol significantly reduced the CaCl(2)-induced contractions in Ca(2+)-free medium. Furthermore, both thymol and carvacrol (300 and 1000 microm) significantly reduced the contraction evoked by phorbol dibutyrate (1 microm), an activator of protein kinase C. Magnitude of this inhibitory effect was enhanced in the presence of the Ca2+ pump inhibitor, thapsigargin (1 microm). At 1000 microm, neither thymol nor carvacrol altered the resting potential of vascular smooth muscle cells. In conclusion, thymol and carvacrol induced an endothelium-independent relaxation in rat isolated aorta, an effect that seems mediated through some mechanisms probably involving a transduction pathway between Ca(2+) release from sarcoplasmic reticulum and/or regulation of the Ca2+ sensitivity of the contractile system. Moreover, it's conceivable that thymol and carvacrol, at low concentrations, block the Ca(2+) influx through the membrane. PMID- 19682088 TI - Combined therapy in the treatment of hypertension. AB - The majority of patients with hypertension need at least two antihypertensive agents to achieve blood pressure (BP) objectives. As current European guidelines for the treatment of arterial hypertension recommend, combined therapy is required when monotherapy fails and as a first-line treatment in certain situations, such as subjects at high or very high cardiovascular risk, markedly elevated BP values, or when lower targets are required (<130/80 mmHg). The advantages of combined therapy are well known and include an earlier and higher antihypertensive efficacy because of complementary mechanisms of action, and a lower incidence of side effects due to the possible compensatory responses and, in many cases, the lower doses used. In the present study, available evidence about the efficacy and tolerability of combined therapy for the treatment of hypertension is updated. PMID- 19682087 TI - Effect of soybean administration on the pharmacokinetics of carbamazepine and omeprazole in rats. AB - Influence of soybean administration on the bioavailability of carbamazepine and omeprazole was studied after single dose administration of soybean (10 g/kg p.o.) or after chronic administration of soybean (50% w/w mixed with normal feed) for 15 days in rats. Carbamazepine was administered orally at a dose of 10 mg/kg and omeprazole at a dose of 20 mg/kg. Soybean decreased the bioavailability of carbamazepine after both single dose and chronic administration. It produced a significant decrease in C(max), T(max), AUC(0-t) of carbamazepine after single dose administration and increased the plasma clearance and V(d) along with decrease in C(max), T(max), AUC(0-t) and AUC(0-infinity) after chronic administration. On the contrary, soybean administration increased the bioavailability of omeprazole by producing an increase in C(max), AUC(0-t) and AUC(0-infinity) and a decrease in V(d) after single dose administration and a decrease in plasma clearance along with increase in C(max), AUC(0-t) and AUC(0 infinity) after chronic administration. The half-life of omeprazole was also increased after both acute and chronic administration of soybean. It was concluded that soybean decreases the bioavailability of carbamazepine and increases the bioavailability of omeprazole after both single dose and chronic administration. PMID- 19682089 TI - Essential requirement for RING finger E3 ubiquitin ligase Hakai in early embryonic development of Drosophila. AB - Hakai is a RING finger type E3 ubiquitin ligase that is highly conserved in metazoans. Mammalian Hakai was shown to bind and ubiquitinate the intracellular domain of E-cadherin, and this activity is implicated in down-regulation of E cadherin during v-Src-induced cellular transformation. To evaluate this model in vivo, we studied the function of the Drosophila homologue of Hakai. In cultured S2 cells, Drosophila Hakai and E-cadherin (Shotgun) formed a complex in a way distinct from the interaction described for mammalian counterparts. Hakai null mutants died during larval stages but this lethality could be offset by a HA tagged Hakai construct. While zygotic Hakai function was dispensable for cell proliferation and differentiation in the wing disc epithelium, maternal Hakai mutants showed a variety of defects in epithelial integrity, including stochastic loss of E-cadherin expression and reduction of aPKC; defects in cell specification and cell migration were also observed. No increase of E-cadherin, however, was observed. Regulation of multiple target proteins under control of Hakai is, therefore, essential for early embryonic morphogenesis in Drosophila. PMID- 19682090 TI - Induction of hyperproliferative fetal megakaryopoiesis by an N-terminally truncated GATA1 mutant. AB - Two GATA1-related leukemias have been described: one is an erythroleukemia that develops in mice as a consequence of diminished expression of wild-type GATA1, whereas the other is an acute megakaryoblastic leukemia (AMKL) that arises in Down syndrome children as a consequence of somatic N-terminal truncation (DeltaNT) of GATA1. We discovered that mice expressing the shortened GATA1 protein (DeltaNTR mice) phenocopies the human transient myeloproliferative disorder (TMD) that precedes AMKL in Down syndrome children. In perinatal livers of the DeltaNTR mutant mice, immature megakaryocytes accumulate massively, and this fraction contains cells that form hyperproliferative megakaryocytic colonies. Furthermore, showing good agreement with the clinical course of TMD in humans, DeltaNTR mutant mice undergo spontaneous resolution from the massive megakaryocyte accumulation concomitant with the switch of hematopoietic microenvironment from liver to bone marrow/spleen. These results thus demonstrate that expression of the GATA1/Gata1 N-terminal deletion mutant per se induces hyperproliferative fetal megakaryopoiesis. This mouse model serves as an important means to clarify how impaired GATA1 function contributes to the multi step leukemogenesis. PMID- 19682091 TI - Fission yeast Ku protein is required for recovery from DNA replication stress. AB - The fundamental function of the conserved Ku70-Ku80 heterodimer is to promote the non-homologous end-joining (NHEJ) pathway in double-strand break repair. Although it is thought that Ku plays several roles other than NHEJ in maintaining chromosomal integrity including telomere protection, these precise functions remain unclear. In this study, we describe a novel role of fission yeast Ku proteins encoded by pku70(+) and pku80(+) genes in dealing with DNA replication stress. In the absence of Rqh1, the fission yeast RecQ helicase, the cells are sensitive to reagents inducing replication stress. pkuDeltarqh1Delta double mutant showed synergistic sensitivities to these reagents. However, this synthetic phenotype was not observed when rqh1Delta mutant was coupled with the deletion of lig4(+) that encodes a ligase essential for NHEJ, indicating that the role of Ku in replication stress is NHEJ independent. pkuDeltarqh1Delta double mutant also showed highly variable copy numbers of rDNA repeats even under unstressed condition. Furthermore, the double mutant exhibited inefficient replication resumption after transient replication stalling. These results suggest the possibility that Ku proteins play an important role in genome integrity recovering replication stress. PMID- 19682092 TI - Prolyl isomerase Pin1 shares functional similarity with phosphorylated CTD interacting factor PCIF1 in vertebrate cells. AB - The carboxy-terminal domain (CTD) of the RNA polymerase II (Pol II) largest subunit undergoes reversible phosphorylation during transcription cycle. The phosphorylated CTD plays critical roles in coordinating transcription with chromatin modification and RNA processing by serving as a scaffold to recruit various proteins. Recently, we identified a novel human WW domain-containing protein PCIF1 as a phosphorylated CTD-interacting factor and demonstrated that PCIF1 negatively modulates Pol II activity in vivo. In the present study, to explore cellular functions of PCIF1, we generated PCIF1-deficient chicken DT40 cell lines. We observed significant up-regulation of WW domain-containing prolyl isomerase Pin1 in two independently established PCIF1-deficient mutant clones. As reconstitution of PCIF1 in the mutants did not reduce Pin1 expression, PCIF1 may not be a negative regulator of Pin1 expression. We assume that Pin1 over expression might suppress defects caused by PCIF1 deficiency in DT40 cells. We furthermore compared PCIF1 and Pin1 for their functional properties and found that these two proteins exhibit most similar target specificity among other CTD binding WW proteins, overlapping subcellular localization and comparative inhibitory effects on transcriptional activation by Pol II in human cultured cells. These results suggest that Pin1 may have overlapping cellular function with PCIF1 in vertebrate cells. PMID- 19682093 TI - An analytical method to determine ground water supply well network designs. AB - An analytical method is provided where the ground water practitioner can quickly determine the size (number of wells) and spacing of a well network capable of meeting a known ground water demand. In order to apply the method, two new parameters are derived that relate theoretical drawdown to the maximum drawdown that is achievable without mining the aquifer. The size of a well network is shown to be proportional to the ground water demand and inversely proportional to the transmissivity and available head. The spacing between wells in a supply well network is shown to be most sensitive to a derived parameter r(HA/3) , which is related to the available head and the propagation of drawdown away from a theoretical well if the total ground water demand was applied to that well. The method can be used to quickly determine the required spacing between wells in well networks of various sizes that are completed in confined aquifers with no leakance. PMID- 19682094 TI - Crescent Moon Spring: a disappearing natural wonder in the Gobi Desert, China. PMID- 19682095 TI - Coupled aquifer-borehole simulation. AB - A model coupling fluid hydraulics in a borehole with fluid flow in an aquifer is developed in this paper. Conservation of momentum is used to create a one dimensional steady-state model of vertical flow in an open borehole combined with radially symmetric flow in an aquifer and with inflow to the well through the wellbore screen. Both laminar and turbulent wellbore conditions are treated. The influence of inflow through the wellbore screen on vertical flow in the wellbore is included, using a relation developed by Siwon (1987). The influence of inflow reduces the predicted vertical variation in head up to 15% compared to a calculation of head losses due to fluid acceleration and the conventional Colebrook-White formulation of friction losses in a circular pipe. The wellbore flow model is embedded into the MODFLOW-2000 ground water flow code. The nonlinear conservation of momentum equations are iteratively linearized to calculate the conductance terms for vertical flow in the wellbore. The resulting simulations agree favorably with previously published results when the model is adjusted to meet the assumptions of the previous coupled models. PMID- 19682096 TI - A 5-year follow-up of older adults residing in long-term care facilities: utilisation of a comprehensive dental programme. AB - OBJECTIVE: This study will compare the clinical outcomes of 139 elders residing in long-term care (LTC) who received dental treatment with those who did not receive care under a comprehensive dental programme over 5 years. BACKGROUND: Numerous studies have documented very poor oral health and limited access to dental care among frail older adults residing in LTC facilities. The University of British Columbia and Providence Healthcare developed a comprehensive dental programme to serve elderly LTC residents within seven Vancouver hospitals. Since 2002, the Geriatric Dentistry Programme has provided annual oral health assessments and access to comprehensive dental care. MATERIAL AND METHODS: A comprehensive oral health assessment was provided using CODE (an index of Clinical Oral Disorders in Elders). A change in oral health status (improvement or worsening) was evaluated by measuring CODE scores including caries and periodontal condition, and other aspects of the dentition. RESULTS: Eighty-three residents received dental treatment of some form over the 5 years, while 56 did not receive any treatment beyond an annual examination. The percentage of residents initially recommended for treatment in 2002 was 97%, which declined to 70-73% after the 3rd year. The percentage of residents treated increased after the first year and remained at 56-72% thereafter. The comparison between CODE scores from baseline and 5 years later showed an improvement for those receiving care (p = 0.02, chi(2) = 7.9, df = 2). CONCLUSION: Within the limitations of this study, residents who did consent and receive care showed an improvement in their oral health status after 5 years. PMID- 19682097 TI - The comparison of marginal bone loss around mandibular overdenture-supporting implants with two different attachment types in a loading period of 36 months. AB - OBJECTIVE: The aim of this study was to assess the influence of attachment types on the marginal bone loss (MBL) around dental implants supporting mandibular overdentures (OVD). BACKGROUND: There are a number of in vitro studies evaluating the influence of several factors on MBL around implants. MATERIAL AND METHODS: Mandibular OVD patients appearing at routine recall sessions consecutively 6, 12, 24 and 36 months after loading were included in the study group. All patients received mandibular OVD with either ball or bar attachments. Measurements were obtained from images of successive radiographs, which were scanned and digitised before, and analysed at *20 magnification. Statistical analyses were utilised in this study to assess the mean marginal bone level changes as well as to explore the potential effect of several parameters such as the cantilever or the attachment type on bone loss. RESULTS: One hundred and twenty-six implants in 51 patients with a mean age of 59.39 +/- 9.99 years were evaluated. There was no statistical significant difference between the distal and mesial bone loss rates of single or splinted attachment types, whereas bone loss rates were statistically higher in cantilever situations. CONCLUSION: Within the limitations of this study, gender, age and diameter of the implants do not play a role in MBL. Length of the implant is an important factor in marginal bone level maintenance. The attachment type for OVD support seems not to influence MBL, but cantilevering of the bars increases bone loss significantly. PMID- 19682098 TI - Exciting but exhausting: experiences with participatory research with chronically ill adolescents. AB - BACKGROUND: Adolescents with chronic conditions are major users of paediatric hospitals, but seldom participate in the evaluation of services or in research. Little is known about the usefulness of the participatory approach in adolescent health research. OBJECTIVE: To evaluate the feasibility, benefits and limitations of a participatory research (PR) project involving chronically ill adolescents as co-researchers. DESIGN SETTING AND PARTICIPANTS: Nine adolescents, aged 15-17 years, acted as co-researchers in a hospital-based PR project. They co-developed an interview protocol and during a disco party held for this purpose interviewed each other and 25 fellow patients (12-19 years). They provided advice on the draft report and participated in the dissemination of the results, but were not involved in the design of the project or analysis of results. RESULTS: Involving adolescents in participatory health research was feasible and appreciated by researchers and youth alike, but had its drawbacks too. The peer-research attracted few participants, the interviews lacked depth and did not yield substantial new insights. Maintaining a high level of participation of the chronically ill co-researchers also proved difficult. CONCLUSIONS: Adolescents with chronic conditions like to have a say in the design and evaluation of hospital services. But their participation as co-researchers demands ample resources from all parties involved without automatically improving research quality. PR does not seem the most effective and efficient way to make services more responsive. We therefore recommend further exploration of other creative and sustainable ways for involving youth in health-care service development and innovation. PMID- 19682099 TI - Queer quit: gay smokers' perspectives on a culturally specific smoking cessation service. AB - BACKGROUND: The prevalence of smoking is high among gay males. The need for culturally specific support has been acknowledged, but little is known about gay men's perspectives on such adapted interventions. OBJECTIVE: To investigate smoking and intention to quit in gay smokers and to explore their attitudes towards a gay-specific smoking cessation programme. DESIGN: Quantitative survey and focus groups. SETTING AND PARTICIPANTS: A total of 325 gay smokers living in Zurich (Switzerland) completed an anonymous survey. Thirteen males participated in two focus groups, theoretically sampled to reflect heterogeneity in terms of age, HIV serostatus and smoking histories. Participants were personally recruited at a variety of events and through advertisements. RESULTS: Responders reported high consumption of cigarettes, and every second man stated that more than half of his gay friends smoke regularly. The majority planned their quit within the next 6 months. Idealizing attitudes towards smoking were very common. Men stated strong preferences towards a culturally adapted cessation programme for gay men. Higher age, high nicotine dependence, intention to quit, smoking stereotypes and fears for weight gain were significant predictors for interest in participation in the programme. Qualitative results indicate that men felt torn between their wish for support, bonding, and community alternatives to the 'smoking gay' environment and fears for failure and loss of reputation. CONCLUSIONS: Gay men reported likely use of a gay-specific intervention. Such interventions may offer support in abstaining from smoking, without abstaining from gay social life. Health-care providers play an important role in communicating the serious threats caused by smoking to gay men. PMID- 19682101 TI - Abacavir and risk of myocardial infarction in HIV-infected patients on highly active antiretroviral therapy: a population-based nationwide cohort study. AB - OBJECTIVE: The aim of the study was to examine whether exposure to abacavir increases the risk for myocardial infarction (MI). DESIGN, SETTING AND SUBJECTS: This was a prospective nationwide cohort study which included all Danish HIV infected patients on highly active antiretroviral therapy (HAART) from 1995 to 2005 (N = 2952). Data on hospitalization for MI and comorbidity were obtained from Danish medical databases. Hospitalization rates for MI after HAART initiation were calculated for patients who used abacavir and those who did not. We used Cox's regression to compute incidence rate ratios (IRR) as a measure of relative risk for MI, while controlling for potential confounders (as separate variables and via propensity score) including comorbidity. MAIN OUTCOME: Relative risk of hospitalization with MI in abacavir users compared with abacavir nonusers. RESULTS: Hospitalization rates for MI were 2.4/1000 person-years (PYR) [95% confidence interval (CI) 1.7-3.4] for abacavir nonusers and 5.7/1000 PYR (95% CI 4.1-7.9) for abacavir users. The risk of MI increased after initiation of abacavir [unadjusted IRR = 2.22 (95% CI 1.31-3.76); IRR adjusted for confounders = 2.00 (95% CI 1.10-3.64); IRR adjusted for propensity score = 2.00 (95% CI 1.07 3.76)]. This effect was also observed among patients initiating abacavir within 2 years after the start of HAART and among patients who started abacavir as part of a triple nucleoside reverse transcriptase inhibitor (NRTI) regimen. CONCLUSIONS: We confirmed the association between abacavir use and increased risk of MI. Further studies are needed to control for potential confounding not measured in research to date. PMID- 19682103 TI - The effects of indoor particles on blood pressure and heart rate among young adults in Taipei, Taiwan. AB - This study aims to evaluate whether indoor particles are associated with elevated blood pressure (BP) and heart rate (HR). We recruited 40 young, healthy students from universities in Taipei. We made four home visits in which we took consecutive 48-h measurements of systolic BP, (SBP) diastolic BP (DBP), and HR in each participant. Particulate matter less than 10 microm in diameter (PM(10)), 2.5 microm in diameter (PM(2.5)), and nitrogen dioxide levels were measured at each participant's home. Participants were asked to keep their windows open during the first two visits, and keep their windows shut during the last two visits. We used linear mixed-effects models to associate BP and HR with indoor air pollutants averaged over 1- to 8-h periods prior to physiological measurements. We found indoor PM(10) and PM(2.5) exposures at 1- to 4-h means were associated with an elevation in SBP, DBP, and HR. Effects of indoor PM(10) and PM(2.5) on BP and HR were greatest during the visits with windows open. During windows-closed visits, participants showed no significant change in BP and HR with indoor PM(10) exposure. We concluded that exposures to infiltrated outdoor particles are associated with short-term increases in BP and HR in young and healthy students. Closing windows can reduce indoor PM concentrations and modify the effect of PM(10) on BP and HR in young adults. PRACTICAL IMPLICATIONS: Particulate matter exposure, high blood pressure (BP) and heart rate (HR) have been reported to be associated with increased risk of cardiovascular morbidity and mortality. Exposure to indoor particles is found to be associated with Elevated BP and HR. Closing windows may reduce indoor particles concentrations and modify the effect of particles on BP and HR in young adults in heavily polluted cities. PMID- 19682100 TI - Long-term (96-week) follow-up of antiretroviral-naive HIV-infected patients treated with first-line lopinavir/ritonavir monotherapy in the MONARK trial. AB - BACKGROUND: The toxicities, cost and complexity of triple combinations warrant the search for other treatment options, such as boosted protease inhibitor (PI) monotherapy. MONotherapy AntiRetroviral Kaletra (MONARK) is the first randomized trial comparing lopinavir/ritonavir monotherapy to triple combination therapy with zidovudine/lamivudine and lopinavir/ritonavir in antiretroviral-naive patients. METHODS: A total of 136 antiretroviral-naive patients, with a CD4 cell count above 100 cells/microL and a plasma HIV RNA below 100,000 HIV-1 RNA copies/mL, were randomized and dosed with either lopinavir/ritonavir monotherapy (n = 83) or lopinavir/ritonavir + zidovudine/lamivudine (n = 53). We focus here on patients in the lopinavir/ritonavir monotherapy arm followed to week 96. The intent-to-treat (ITT) analysis initially involved all patients randomized to lopinavir/ritonavir monotherapy (n = 83), and then focused on patients who had an HIV RNA < 50 copies/mL at week 48 (n = 56). RESULTS: At week 96, 39 of 83 patients (47%) had HIV RNA < 50 copies/mL, five of 83 had HIV RNA between 50 and 400 copies/mL, and three of 83 had HIV RNA > 400 copies/mL. Focusing on the 56 patients with an HIV RNA < 50 copies/mL at week 48, 38 of 56 patients (68%) had a sustained HIV RNA < 50 copies/mL to week 96. To week 96, a total of 28 patients (34%) had discontinued the study treatment. In addition, the allocated treatment was changed for seven patients. PI-associated resistance mutations were evident in five of 83 patients in the monotherapy arm from baseline to week 96. CONCLUSION: By ITT analysis, 39 of the 83 patients initially randomized to lopinavir/ritonavir monotherapy had HIV RNA < 50 copies/mL at week 96. The occurrence in some patients of low-level viraemia (50-500 copies/mL) may increase the risk of drug resistance. First-line lopinavir/ritonavir monotherapy cannot be systematically recommended. PMID- 19682102 TI - Emergency department utilization among HIV-infected patients in a multisite multistate study. AB - OBJECTIVE: The aim of this study was to examine Emergency Department (ED) utilization and clinical and sociodemographic correlates of ED use among HIV infected patients. METHODS: During 2003, 951 patients participated in face-to face interviews at 14 HIV clinics in the HIV Research Network. Respondents reported the number of ED visits in the preceding 6 months. Using logistic regression, we identified factors associated with visiting the ED in the last 6 months and admission to the hospital from the ED. RESULTS: Thirty-two per cent of respondents reported at least one ED visit in the last 6 months. In multivariate analysis, any ED use was associated with Medicaid insurance, high levels of pain (the third or fourth quartile), more than seven primary care visits in the last 6 months, current or former illicit drug use, social alcohol use and female gender. Of those who used ED services, 39% reported at least one admission to the hospital. Patients with pain in the highest quartile reported increased admission rates from the ED as did those who made six or seven primary care visits, or more than seven primary care visits vs. three or fewer. CONCLUSIONS: The likelihood of visiting the ED has not diminished since the advent of highly active antiretroviral therapy (HAART). More ED visits are to treat illnesses not related to HIV or injuries than to treat direct sequelae of HIV infection. With the growing prevalence of people living with HIV infection, the numbers of HIV infected patients visiting the ED may increase, and ED providers need to understand potential complications produced by HIV disease. PMID- 19682104 TI - An association between floor vacuuming and dust-mite and serum eosinophil cationic protein in young asthmatics. AB - Although vacuum cleaning is recommended to reduce allergen levels and improve asthma and allergic rhinitis symptoms, some studies suggest it may increase allergen load in homes. We conducted a cross-sectional study to determine if home floor vacuuming was associated with sensitization to dust-mites and cockroaches, and serum eosinophil cationic protein (ECP), a biomarker for atopy, in 102 physician-diagnosed spirometry-confirmed asthmatics. We collected data on floor type, floor cleaning method and frequency, asthma severity, allergy medications used, serum ECP and skin prick tests (SPT) to three dust-mites [Dermatophagoides pteronyssisinus (Der p), Dermatophagoides farinae (Der f) and Blomia tropicalis (Blo t)] and two cockroaches [Periplanata americana (Per a) and Blatella germanica (Bla g)]. Those who vacuumed had increased sensitization to three dust mites [adjusted ORs (95%CI) = Der p: 26.6 (1.8-405.2); Der f: 44.8 (3.2-620.9); Blo t: 14.1 (1.8-108.1)] but not to cockroaches, adjusted for cleaning frequency and other methods of floor cleaning. Subjects who vacuumed their floor had higher levels of serum ECP than those who did not [adjusted median difference (95%CI): 9.4 (1.1-17.7)], adjusted for use of nasal corticosteroids among those with allergic rhinitis. Vacuuming is associated with increased sensitization to dust mite allergens and higher serum ECP. PRACTICAL IMPLICATIONS: We found an association between floor vacuuming and increased sensitization to dust-mite allergens and higher levels of an atopy biomarker. Current recommendations to use vacuuming to control allergen exposure and allergic conditions may need to be reconsidered until further studies are performed. PMID- 19682105 TI - Female urology: what method will you select? PMID- 19682106 TI - Lateral view dissection of the prostato-urethral junction to reduce positive apical margin in laparoscopic radical prostatectomy. AB - OBJECTIVE: To assess the impact of lateral view apical dissection in laparoscopic radical prostatectomy (LRP) on the reduction of positive surgical margin rates and recovery of postoperative continence. METHODS: One hundred and forty-four consecutive patients underwent LRP from October 2004 to March 2008. Lateral view dissection of the prostato-urethral junction was conducted in 76 of them (Group 2). Standard dissection was used in the remaining patients (Group 1). The effect of this technical modification on the reduction of positive surgical margin rates and postoperative recovery of urinary continence was assessed in the two groups. RESULTS: Overall, the incidence of positive margins decreased from 23 (35.9%) in Group 1 to 16 cases (21.9%) in Group 2 (P = 0.07). Positive margin rates in pT2 decreased from 30.6% to 6.5% (P = 0.006). Apical and dorso-apical margins were reduced from 26.5% to 4.3% (P = 0.009) and from 10.2% to 0% (P < 0.001), respectively. Postoperative recovery of urinary continence improved significantly, with a pad-free rate over the first 3 months of 55.9% in Group 1 vs 71.7% in Group 2 (P = 0.01). Multivariate logistic regression analysis showed this modified surgical technique to predict a lower rate of positive margins. CONCLUSION: Lateral view dissection of the prostato-urethral junction is an easily applicable technical modification. It provides better visualization of apical anatomy substantially contributing to the reduction of positive surgical margin rates, especially at the level of prostatic apex. PMID- 19682107 TI - A case of renal mucinous tubular and spindle cell carcinoma. AB - A 33-year-old man was hospitalized for treatment of a left renal tumor. The radiological findings were consistent with those of a left renal cell carcinoma (RCC). Subsequently, a radical nephrectomy was carried out. Macroscopic examination showed that a well-demarcated tumor measuring 2.9 x 2.6 x 2.5 cm was present in the middle portion of the resected kidney. The cut surface of the tumor was grayish-white in color. Pathological examination of the resected specimen showed a mucinous tubular and spindle cell carcinoma of the kidney (MTSCC-K). MTSCC-K is a low-grade renal epithelial neoplasm that has recently been recognized as a specific entity in the World Health Organization 2004 classification of RCC. To our knowledge, 17 cases of MTSCC-K in Japan have been reported by Japanese investigators. To avoid administration of excessive adjuvant treatment to patients, pathologists and urologists should consider this newly recognized low-grade malignancy when diagnosing renal tumors. PMID- 19682108 TI - Enhanced VDAC-cored VRAC in NE-LNCaP cells points to an involvement of eukaryotic porin in the extrinsic apoptotic pathway. PMID- 19682110 TI - Fourth joint meeting of the American Urological Association and the Japanese Urological Association Specialty Society program at the 104th annual meeting of the American Urological Association at Chicago 2009. AB - We are heartily grateful for the warm support of all of the people concerned, including the moderators and panelists of both societies for giving us the opportunity to hold the 4th American Urological Association/Japanese Urological Association (AUA/JUA) Joint Meeting, held once again at the 104th Annual Meeting of the American Urological Association (25-30 April 2009, Chicago, Illinois, USA). 2009 is a memorable year, being the start of new collaborations between AUA and JUA. The JUA in collaboration with AUA is promoting an academic exchange program whereby outstanding and promising Japanese and American junior faculty members will be given the opportunity to work in the USA and Japan for one month. The program not only allows the sharing of knowledge and experience, but is designed to foster a closer alliance between the AUA and JUA, and assists in identifying future leaders within both organizations. The JUA will have an exhibit booth at the AUA annual meeting, promoting our new joint activities. The Journal of Urology and International Journal of Urology will share reviewers. The JUA will participate in developing AUA guidelines. With all of these activities, the JUA hopes it will provide greater opportunities to young Japanese urologists to participate in educational projects in the US. We would like to thank Professor Robert C. Flanigan, the Secretary General of AUA, Professor Glenn M. Preminger, the Chairman of the AUA Office of Education and the staff of AUA and JUA for supporting our program. We hope to keep holding the joint meeting and have plenty of ideas on themes and forums. We believe that this international program helps to establish a closer relationship between JUA and AUA in the scientific field. PMID- 19682112 TI - Validation and comparison of two frailty indexes: The MOBILIZE Boston Study. AB - OBJECTIVES: To validate two established frailty indexes and compare their ability to predict adverse outcomes in a diverse, elderly, community-dwelling sample of men and women. DESIGN: Prospective observational study. SETTING: A diverse defined geographic area of Boston. PARTICIPANTS: Seven hundred sixty-five community-dwelling participants in the Maintenance of Balance, Independent Living, Intellect, and Zest in the Elderly Boston Study. MEASUREMENTS: Two published frailty indexes, recurrent falls, disability, overnight hospitalization, emergency department (ED) visits, chronic medical conditions, self-reported health, physical function, cognitive ability (including executive function), and depression. One index was developed from the Study of Osteoporotic Fractures (SOF) and the other from the Cardiovascular Health Study (CHS). RESULTS: The SOF frailty index classified 77.1% as robust, 18.7% as prefrail, and 4.2% as frail. The CHS frailty index classified 51.2% as robust, 38.8% as prefrail, and 10.0% as frail. Both frailty indexes (SOF; CHS) were similar in their ability to predict key geriatric outcomes such as recurrent falls (hazard ratio (HR)(frail)=2.2, 95% confidence interval (CI)=1.2-4.0; HR(frail)=1.9, 95% CI=1.2-3.1), overnight hospitalization (odds ratio (OR)(frail)=3.5, 95% CI=1.5 8.0; OR(frail)=4.4, 95% CI=2.4-8.2), ED visits (OR(frail)=3.5, 95% CI=1.4,8.8; OR(frail)=3.1, 95% CI=1.6-5.9), and disability (OR(frail)=5.4, 95% CI=2.3-12.3; OR(frail)=7.7, 95% CI=4.0-14.7), as well as chronic medical conditions, physical function, cognitive ability, and depression. CONCLUSION: Two established frailty indexes were validated using an independent elderly sample of diverse men and women; both indexes were good at distinguishing geriatric conditions and predicting recurrent falls, overnight hospitalization, and ED visits according to level of frailty. Although both indexes are good measures of frailty, the simpler SOF index may be easier and more practical in a clinical setting. PMID- 19682111 TI - Use of the late-life function and disability instrument to assess disability in major depression. AB - OBJECTIVES: To determine whether there was greater disability in subjects with depression than in those without, the correlation between disability and depression severity and quality of life, and whether improvement in disability after antidepressant pharmacotherapy was greater in those who responded to antidepressant treatment. DESIGN: Disability in subjects with and without depression from two different studies was compared for 22 weeks. Correlations were performed for the subjects with depression between disability and depression, anxiety, health-related quality of life (HRQOL), and medical comorbidity. T-tests were used to compare disability between subjects who did and did not respond to antidepressant treatment and change in disability after pharmacotherapy. SETTING: Late-life depression research clinic. PARTICIPANTS: The 313 subjects were recruited from primary care and the community and were aged 60 and older; 244 subjects were participants in a depression treatment protocol, and 69 subjects without depression participated in a separate longitudinal observational study of the mental and cognitive health of depression-free older adults. MEASUREMENTS: The Late-Life Function and Disability Instrument (LL-FDI), a measure of instrumental activity of daily living, personal role, and social role functioning. RESULTS: Subjects with depression scored lower than controls for domains measuring limitation (can do) and frequency (does do) of activities. Both disability domains correlated with depression severity, anxiety, HRQOL, and cognition. Disability improved with antidepressant treatment; for partial responders who continued to receive higher-dose antidepressant treatment out to 22 weeks, there was continued improvement, although not to the level of comparison subjects without depression. CONCLUSION: The LL-FDI appears to discriminate subjects with depression from those without, correlates with depression severity, and demonstrates sensitivity to antidepressant treatment response. We recommend further investigation of the LL-FDI and similar disability instruments for assessing depression-related disability. PMID- 19682113 TI - High oxidative stress is correlated with frailty in elderly chinese. AB - OBJECTIVES: To evaluate the relationship between oxidative stress and frailty in elderly people. DESIGN: Cross-sectional study. SETTING: Community and hospital based outpatient clinic. PARTICIPANTS: Ninety participants aged 65 and older. MEASUREMENTS: Frailty status was determined according to the presence of weak handgrip strength, weight loss, slow walking speed, exhaustion, and low activity level and was classified as frail (> or =3 criteria), prefrail (1 or 2 criteria), or robust (0 criteria). An oxidative stress marker (serum 8-hydroxy-2' deoxyguanosine, 8-OHdG), metabolic markers (body mass index, waist-hip ratio, serum lipids, glucose, and albumin), an inflammatory marker (serum high sensitivity C-reactive protein, hs-CRP), demographic information, and comorbidities (diabetes mellitus, hypertension, congestive heart failure, osteoarthritis, overweight or obesity, impaired fasting plasma glucose, renal insufficiency, and depression) were assessed. RESULTS: Of the 90 participants, 21 (23.3%) were frail, 56 (62.2%) were prefrail, and 13 (14.4%) were robust. Frail subjects had higher median (range) serum 8-OHdG (2.5 ng/mL (1.5-6.2 ng/mL) vs 2.3 ng/mL (0.5-8.1 ng/mL) and 1.0 ng/mL (0.5-5.3 ng/mL)) and serum hs-CRP (2.5 mg/L (0.3-32.1 mg/L) vs 1.8 mg/L (0.3-50.5 mg/L) and 1.7 mg/L (0.3-4.0 mg/L)) levels, lower mean+/-standard deviation serum albumin levels (4.1+/-0.4 g/dL vs 4.4+/-0.4 g/dL and 4.6+/-0.2 g/dL) and higher mean waist-hip ratios (0.96+/-0.11 vs 0.91+/ 0.07 and 0.89+/-0.05)) than prefrail and robust subjects, respectively (P<.05 for all). In multivariable regression analysis, high serum 8-OHdG level was still significantly associated with frailty after adjusting for age, smoking status, comorbidities, waist-hip ratio, serum albumin level, and hs-CRP level. CONCLUSION: High oxidative stress, characterized by high serum 8-OHdG level, was independently associated with frailty in the selected sample of elderly Chinese. PMID- 19682114 TI - Yoga decreases kyphosis in senior women and men with adult-onset hyperkyphosis: results of a randomized controlled trial. AB - OBJECTIVES: To assess whether a specifically designed yoga intervention can reduce hyperkyphosis. DESIGN: A 6-month, two-group, randomized, controlled, single-masked trial. SETTING: Community research unit. PARTICIPANTS: One hundred eighteen women and men aged 60 and older with a kyphosis angle of 40 degrees or greater. Major exclusions were serious medical comorbidity, use of assistive device, inability to hear or see adequately for participation, and inability to pass a physical safety screen. INTERVENTION: The active treatment group attended hour-long yoga classes 3 days per week for 24 weeks. The control group attended a monthly luncheon and seminar and received mailings. MEASUREMENTS: Primary outcomes were change (baseline to 6 months) in Debrunner kyphometer-assessed kyphosis angle, standing height, timed chair stands, functional reach, and walking speed. Secondary outcomes were change in kyphosis index, flexicurve kyphosis angle, Rancho Bernardo Blocks posture assessment, and health-related quality of life (HRQOL). RESULTS: Compared with control participants, participants randomized to yoga experienced a 4.4% improvement in flexicurve kyphosis angle (P=.006) and a 5% improvement in kyphosis index (P=.004). The intervention did not result in statistically significant improvement in Debrunner kyphometer angle, measured physical performance, or self-assessed HRQOL (each P>.1). CONCLUSION: The decrease in flexicurve kyphosis angle in the yoga treatment group shows that hyperkyphosis is remediable, a critical first step in the pathway to treating or preventing this condition. Larger, more-definitive studies of yoga or other interventions for hyperkyphosis should be considered. Targeting individuals with more-malleable spines and using longitudinally precise measures of kyphosis could strengthen the treatment effect. PMID- 19682115 TI - Estimating the quantity and economic value of family caregiving for community dwelling older persons in the last year of life. AB - OBJECTIVES: To estimate the quantity and economic value of informal care provided to older persons during their final year of life in the community. DESIGN: Retrospective analysis of publicly available nationally representative survey data. SETTING: This retrospective study used data from the Health and Retirement Study, a nationally representative, longitudinal study of community-dwelling older people. PARTICIPANTS: Older people who died between 2000 and 2002. MEASUREMENTS: Data were extracted from the 2002 "exit survey" and linked with characteristics of caregivers from the helper file. Ordinary least squares regression was used to estimate hours of informal caregiving for community dwelling older people (N=990). Adjusted hours were multiplied by the 2002 national average home aide wage (9.16 USD per hour). Sensitivity tests were performed using the 10th percentile wage rate (6.56 USD) and 90th percentile wage rate (12.34 USD). RESULTS: Older people who died in the community received on average 65.8 hours per week of informal care in the last year of life. The estimated economic value ranges from 22,514 USD to 42,351 USD, which is equivalent to the annual direct replacement cost with a home aide in 2002. CONCLUSION: Family members provide substantial assistance during the last year of life for older people who die in the community. If the informal care provided in the last year of life is replaced with a home aide, the total economic value for the United States would be approximately 1.4 billion USD (in 2002). PMID- 19682116 TI - Allostatic load and frailty in older adults. AB - OBJECTIVES: To examine the association between allostatic load (AL), an index of multisystem physiological dysregulation, and frailty development over a 3-year follow-up in a sample of older adults. DESIGN: Longitudinal cohort study. SETTING: Community. PARTICIPANTS: High-functioning men and women aged 70 to 79 at study entry. MEASUREMENTS: Multisystem physiological dysregulation, or AL, was assessed according to 13 biomarkers of cardiovascular, endocrine, immune, and metabolic function. An AL score was computed as the total number of biomarkers for which participant values fell into high-risk biomarker quartiles. Frailty status (not frail, intermediate frail, frail) was determined according to the total number of five indicators of frailty: weight loss, exhaustion, weak grip, slow gait, and low physical activity. The association between level of AL at baseline and frailty status 3 years later was examined using ordinal logistic regression in 803 participants not frail at baseline. RESULTS: In a multivariable model adjusting for sociodemographic, health, and behavioral characteristics, each 1-unit increase in AL at baseline was associated with a 10% greater likelihood of frailty at the 3-year follow-up (cumulative adjusted odds ratio=1.10, 95% confidence interval=1.03-1.19). CONCLUSION: These findings support the hypothesis that dysregulation across multiple physiological systems is associated with greater risk of frailty. Greater levels of multisystem physiological dysregulation may serve as a warning sign of frailty development in later life. PMID- 19682117 TI - Nonagenarian siblings and their offspring display lower risk of mortality and morbidity than sporadic nonagenarians: The Leiden Longevity Study. AB - OBJECTIVES: To compare the risk of mortality of nonagenarian siblings with that of sporadic nonagenarians (not selected on having a nonagenarian sibling) and to compare the prevalence of morbidity in their offspring with that of the offsprings' partners. DESIGN: Longitudinal (mortality risk) and cross-sectional (disease prevalence). SETTING: Nationwide sample. PARTICIPANTS: The Leiden Longevity Study consists of 991 nonagenarian siblings derived from 420 Caucasian families, 1,365 of their offspring, and 621 of the offsprings' partners. In the Leiden 85-plus Study, 599 subjects aged 85 were included, of whom 275 attained the age of 90 (sporadic nonagenarians). MEASUREMENTS: All nonagenarian siblings and sporadic nonagenarians were followed for mortality (with a mean+/-standard deviation follow-up time of 2.7+/-1.4 years and 3.0+/-1.5 years, respectively). Information on medical history and medication use was collected for offspring and their partners. RESULTS: Nonagenarian siblings had a 41% lower risk of mortality (P<.001) than sporadic nonagenarians. The offspring of nonagenarian siblings had a lower prevalence of myocardial infarction (2.4% vs 4.1%, P=.03), hypertension (23.0% vs 27.5%, P=.01), diabetes mellitus (4.4% vs 7.6%, P=.004), and use of cardiovascular medication (23.0% vs 28.9%, P=.003) than their partners. CONCLUSION: The lower mortality rate of nonagenarian siblings and lower prevalence of morbidity in their middle-aged offspring reinforce the notion that resilience against disease and death have similar underlying biology that is determined by genetic or familial factors. PMID- 19682118 TI - Effect of influenza vaccination of nursing home staff on mortality of residents: a cluster-randomized trial. AB - OBJECTIVES: To evaluate the effect of staff influenza vaccination on all-cause mortality in nursing home residents. DESIGN: Pair-matched cluster-randomized trial. SETTING: Forty nursing homes matched for size, staff vaccination coverage during the previous season, and resident disability index. PARTICIPANTS: All persons aged 60 and older residing in the nursing homes. INTERVENTION: Influenza vaccine was administered to volunteer staff after a face-to-face interview. No intervention took place in control nursing homes. MEASUREMENTS: The primary endpoint was total mortality rate in residents from 2 weeks before to 2 weeks after the influenza epidemic in the community. Secondary endpoints were rates of hospitalization and influenza-like illness (ILI) in residents and sick leave from work in staff. RESULTS: Staff influenza vaccination rates were 69.9% in the vaccination arm versus 31.8% in the control arm. Primary unadjusted analysis did not show significantly lower mortality in residents in the vaccination arm (odds ratio=0.86, P=.08), although multivariate-adjusted analysis showed 20% lower mortality (P=.02), and a strong correlation was observed between staff vaccination coverage and all-cause mortality in residents (correlation coefficient=-0.42, P=.007). In the vaccination arm, significantly lower resident hospitalization rates were not observed, but ILI in residents was 31% lower (P=.007), and sick leave from work in staff was 42% lower (P=.03). CONCLUSION: These results support influenza vaccination of staff caring for institutionalized elderly people. PMID- 19682119 TI - Targeting nursing homes under the Quality Improvement Organization program's 9th statement of work. AB - In the Quality Improvement Organization (QIO) program's latest Statement of Work, the Centers for Medicare and Medicaid Services (CMS) is targeting its nursing home activities toward facilities that perform poorly on two quality measures pressure ulcers and restraint use. The designation of target facilities is a shift in strategy for CMS and a direct response to criticism that QIO program resources were not being targeted effectively to facilities or clinical areas that most needed improvement. Using administrative data, this article analyzes implications of using narrowly defined criteria to identify facilities that need improvement, particularly in light of considerable evidence showing that nursing home quality is multidimensional and may change over time. The analyses show that one in four facilities is targeted for improvement nationally but that approximately half of some states' facilities are targeted while other states have almost none targeted. The analyses also convey deeper limitations to using threshold values on individual measures to identify poorly performing homes. Target facilities can be among the top performers on a range of other quality measures, and their performance on targeted measures themselves may change over time. The implication of these features is that a very different group of facilities would have been chosen had the QIO program targeted other measures or examined performance at a different point in time. Ultimately, CMS has chosen a blunt instrument to identify poorly performing nursing homes, and supplemental strategies-such as soliciting input from state survey agencies and more closely aligning quality improvement and quality assurance efforts-should be considered to address potential limitations. PMID- 19682120 TI - Understanding advance care planning as a process of health behavior change. AB - OBJECTIVES: To explore whether models of health behavior change can help to inform interventions for advance care planning (ACP). DESIGN: Qualitative cross sectional study. SETTING: Community. PARTICIPANTS: Sixty-three community-dwelling persons aged 65 and older and 30 caregivers with experience as surrogate decision makers. MEASUREMENTS: In focus groups conducted separately with older persons and caregivers, participants were asked to discuss ways they had planned for future declines in health and why they had or had not engaged in such planning. Transcripts were analyzed using grounded theory. RESULTS: Four themes illustrated the potential of applying models of health behavior change to improve ACP. (1) Participants demonstrated variable readiness to engage in ACP and could be in different stages of readiness for different components of ACP, including consideration of treatment goals, completion of advance directives, and communication with families and physicians. (2) Participants identified a wide range of benefits of and barriers to ACP. (3) Participants used a variety of processes of change to progress through stages of readiness, and ACP was only one of a broader set of behaviors that participants engaged in to prepare for declines in their health or for death. (4) Experience with healthcare decision making for loved ones was a strong influence on perceptions of susceptibility and engagement in ACP. DISCUSSION: The variability in participants' readiness, barriers and benefits, perceptions of susceptibility, and use of processes to increase readiness for participating in each component of ACP suggests the utility of customized, stage-specific interventions based on individualized assessments to improve ACP. PMID- 19682121 TI - The underrecognized epidemic of low mobility during hospitalization of older adults. AB - OBJECTIVES: To examine the proportion of time spent in three levels of mobility (lying, sitting, and standing or walking) by a cohort of hospitalized older veterans as measured by validated wireless accelerometers. DESIGN: A prospective, observational cohort study. SETTING: One hundred fifty-bed Department of Veterans Affairs hospital. PARTICIPANTS: Forty-five hospitalized medical patients, aged 65 and older who were not delirious, did not have dementia, and were able to walk in the 2 weeks before admission were eligible. MEASUREMENTS: Wireless accelerometers were attached to the thigh and ankle of patients for the first 7 days after admission or until hospital discharge, whichever came first. The mean proportion of time spent lying, sitting, and standing or walking was determined for each hour after hospital admission using a previously validated algorithm. RESULTS: Forty-five male patients (mean age 74.2) with a mean length of stay of 5.1 days generated 2,592 one-hour periods of data. A baseline functional assessment indicated that 35 (77.8%) study patients were willing and able to walk a short distance independently. No patient remained in bed the entire measured hospital stay, but on average, 83% of the measured hospital stay was spent lying in bed. The average amount of time that any one individual spent standing or walking ranged from a low of 0.2% to a high of 21%, with a median of 3%, or 43 minutes per day. CONCLUSION: This is the first study to continuously monitor mobility levels early during a hospital stay. On average, older hospitalized patients spent most of their time lying in bed, despite an ability to walk independently. PMID- 19682122 TI - Pain, functional limitations, and aging. AB - OBJECTIVES: To examine the relationship between functional limitations and pain across a spectrum of age, ranging from mid life to advanced old age. DESIGN: Cross-sectional study. SETTING: The 2004 Health and Retirement Study (HRS), a nationally representative study of community-living persons aged 50 and older. PARTICIPANTS: Eighteen thousand five hundred thirty-one participants in the 2004 HRS. MEASUREMENTS: Participants who reported that they were often troubled by pain that was moderate or severe most of the time were defined as having significant pain. For each of four functional domains, subjects were classified according to their degree of functional limitation: mobility (able to jog 1 mile, able to walk several blocks, able to walk one block, unable to walk one block), stair climbing (able to climb several flights, able to climb one flight, not able to climb a flight), upper extremity tasks (able to do 3, 2, 1, or 0), and activity of daily living (ADL) function (able to do without difficulty, had difficulty but able to do without help, need help). RESULTS: Twenty-four percent of participants had significant pain. Across all four domains, participants with pain had much higher rates of functional limitations than subjects without pain. Participants with pain were similar in terms of their degree of functional limitation to participants 2 to 3 decades older. For example, for mobility, of subjects aged 50 to 59 without pain, 37% were able to jog 1 mile, 91% were able to walk several blocks, and 96% were able to walk one block without difficulty. In contrast, of subjects aged 50 to 59 with pain, 9% were able to jog 1 mile, 50% were able to walk several blocks, and 69% were able to walk one block without difficulty. Subjects aged 50 to 59 with pain were similar in terms of mobility limitations to subjects aged 80 to 89 without pain, of whom 4% were able to jog 1 mile, 55% were able to walk several blocks, and 72% were able to walk one block without difficulty. After adjustment for demographic characteristics, socioeconomic status, comorbid conditions, depression, obesity, and health habits, across all four measures, participants with significant pain were at much higher risk for having functional limitations (adjusted odds ratio (AOR)=2.85, 95% confidence interval (CI)=2.20-3.69, for mobility; AOR=2.84, 95% CI=2.48-3.26, for stair climbing; AOR=3.96, 95% CI=3.43-4.58, for upper extremity tasks; and AOR=4.33; 95% CI=3.71-5.06, for ADL function). CONCLUSION: Subjects with pain develop the functional limitations classically associated with aging at much earlier ages. PMID- 19682124 TI - Renal function in older hospital patients is more accurately estimated using the Cockcroft-Gault formula than the modification diet in renal disease formula. AB - OBJECTIVES: To compare the accuracy of the two most popular creatinine clearance (CrCl) estimation formulae (Cockcroft-Gault (CG) and Modification Diet in Renal Disease (MDRD)) in older hospitalized patients. DESIGN: Prospective, cross sectional, observational study. SETTING: Two hospital geriatric wards. PARTICIPANTS: Consecutive patients aged 70 and older with an indwelling urinary catheter for the purpose of care. MEASUREMENTS: CrCl was determined according to three methods: measured CrCl from plasma and urine creatinine and 24-hour urine volume, CG (CG-CrCl), and MDRD (MDRD-CrCl). Results were expressed as median and interquartile range (IQR). Moderate and severe renal impairment were defined as a CrCl between 30.0 and 59.9 mL/min and less than 30.0 mL/min, respectively. RESULTS: One hundred twenty-one patients were included (46% male). Mean age was 86.1+/-6.7 (range 72-100). Median measured CrCl was 43.8 mL/min (IQR 33.6-61.1 mL/min), CG-CrCl was 40.9 mL/min (IQR 31.0-52.6 mL/min), and MDRD-CrCl was 61.3 mL/min (IQR 49.4-77.0 mL/min). The biases of CG-CrCl and MDRD were -3.5+/-22.5 and 20.1+/-28.2, respectively (P<.001). Misclassification of renal impairment (absent/moderate/severe) occurred in 33% of patients according to CG-CrCl, and concordance was mild to moderate (kappa=0.50). Misclassification occurred in 50% of patients according to MDRD-CrCl, and concordance was poor (kappa=0.33). Bias was significantly related to bed confinement for both formulae and to plasma creatinine for MDRD. CONCLUSION: In elderly hospitalized patients, CG slightly underestimates CrCl, and MDRD strongly overestimates it. CG gave a better prediction of measured CrCl than MDRD. PMID- 19682123 TI - Development and pilot testing of computerized order entry algorithms for geriatric problems in nursing homes. AB - OBJECTIVES: To develop order entry algorithms for five common nursing home problems and to test their acceptance, use, and preliminary effect on nine quality indicators and resource utilization. DESIGN: Pre-post, quasi-experimental study. SETTING: Two Department of Veterans Affairs nursing homes. PARTICIPANTS: Randomly selected residents (N=265) with one or more target conditions and 42 nursing home providers. INTERVENTION: Expert panels developed computerized order entry algorithms based on clinical practice guidelines. Each was displayed on a single screen and included an array of diagnostic and treatment options and means to communicate with the interdisciplinary team. MEASUREMENTS: Medical records were abstracted for the 6 months before and after deployment for quality indicators and resource utilization. RESULTS: Despite positive provider attitudes toward the computerized order entry algorithms, their use was infrequent and varied according to condition: falls (73.0%), fever (9.0%), pneumonia (8.0%), urinary tract infection (7.0%), and osteoporosis (3.0%). In subjects with falls, trends for improvements in quality measures were observed for six of the nine measures: measuring orthostatic blood pressure (17.5-30.0%, P=.29), reducing neuroleptics (53.8-75.0%, P=.27), reducing sedative-hypnotics (16.7-50.0%, P=.50), prescription of calcium (22.5-32.5%, P=.45), vitamin D (20.0-35.0%, P=.21), and external hip protectors (25.0-47.5%, P=.06). Little improvement was observed in the other conditions (documentation of vital signs, physical therapy referrals, or reduction of benzodiazepines or antidepressants). There was no change in resource utilization. CONCLUSION: Computerized order entry algorithms were used infrequently, except for falls. Further study may determine whether their use leads to improved care. PMID- 19682126 TI - The use of physical restraints in home care in the Netherlands. AB - OBJECTIVES: To gain more knowledge of the application of physical restraints to restrict, restrain, or prevent movement by elderly people living at home. DESIGN: Survey. SETTING: Nursing staff in Dutch home care. PARTICIPANTS: One hundred fifty-seven nursing staff in home care (registered nurses and certified nursing assistants) from a randomly selected and nationally representative panel returned the questionnaire (response rate 72%). MEASUREMENTS: A structured questionnaire on the use of physical restraints. RESULTS: Four of every five nursing staff members have applied physical restraints. The use of bed rails, putting the client in a deep chair or using a chair with a table, and locking doors to prevent wandering were most frequently applied, often at the request of the client or his or her family. Protection of the client is the reason most frequently given for these actions. Almost all respondents (94%) know of no alternatives, nor does consensus exist on what is considered to be a physical restraint. CONCLUSION: Guidelines are necessary regarding the course of action to be taken when a client is in danger of hurting him- or herself. Further education on and due consideration of the use of physical restraints in home care are also required. PMID- 19682125 TI - The role of nursing home admission and dementia status on care for diabetes mellitus. AB - OBJECTIVES: To study the role of nursing home (NH) admission and dementia status on the provision of five procedures related to diabetes mellitus. DESIGN: Retrospective cohort study using data from a large prospective study in which an expert panel determined the prevalence of dementia. SETTING: Fifty-nine Maryland NHs. PARTICIPANTS: Three hundred ninety-nine new admission NH patients with diabetes mellitus. MEASUREMENTS: Medicare administrative claims records matched to the NH medical record data were used to measure procedures related to diabetes mellitus received in the year before NH admission and up to a year after admission (and before discharge). Procedures included glycosylated hemoglobin, fasting blood glucose, dilated eye examination, lipid profile, and serum creatinine. RESULTS: For all but dilated eye examinations, higher rates of procedures related to diabetes mellitus were seen in the year after NH admission than in the year before. Residents without dementia received more procedures than those with dementia, although this was somewhat attenuated after controlling for demographic, health, and healthcare utilization variables. Persons without dementia experience greater increases in procedure rates after admission than those with dementia. CONCLUSION: The structured environment of care provided by the NH may positively affect monitoring procedures provided to elderly persons with diabetes mellitus, especially those without dementia. Medical decisions related to the risks and benefits of intensive treatment for diabetes mellitus to patients of varying frailty and expected longevity may lead to lower rates of procedures for residents with dementia. PMID- 19682128 TI - Baseline and follow-up characteristics of participants and nonparticipants in a randomized clinical trial of multifactorial fall prevention in Denmark. AB - OBJECTIVES: To address the external validity of a trial of multifactorial fall prevention through an analysis of differences between participants and nonparticipants regarding socioeconomic and morbidity variables. DESIGN: Analysis of nonresponse in a randomized clinical trial. SETTING: Geriatric outpatient department. PARTICIPANTS: One thousand one hundred five community-dwelling adults aged 65 and older who had sustained at least one injurious fall. MEASUREMENTS: Marital status, housing tenure, income, comorbidity, hospitalization, fractures, and drug use before invitation to participate in the trial. Fractures, hospitalization and death were measured for 6 months of follow-up. RESULTS: Four hundred forty-seven responding nonparticipants and 266 nonresponding nonparticipants were compared with 392 participants in the trial. Lower income (odds ratio (OR)=2.38, 95% confidence interval (CI)=1.28-4.28) and more days of hospitalization during the previous 5 years (OR=1.96, 95% CI=1.15-3.33) predicted responding nonparticipation; independent predictors of being a nonresponding nonparticipant were unmarried status (OR=2.0, 95% CI=1.36-2.94), lower income (OR=4.74, 95% CI=2.30-9.78), more days of hospitalization (OR=3.49, 95% CI=1.99 6.11), and prior fractures (OR=1.56, 95% CI=1.02-2.38). Nonresponding nonparticipants were significantly more likely to die (OR=12.99, 95% CI=1.6 105.6) or be hospitalized (OR=2.66, 95% CI=1.7-4.1) than participants during 6 months of follow-up. CONCLUSION: Nonresponding nonparticipants of a trial of multifactorial fall prevention differed significantly from participants in terms of socioeconomic and morbidity variables and were more likely to be hospitalized or die during 6 months of follow-up. Because of the differences between the two populations, it is questionable whether results from this randomized trial can be generalized to people potentially eligible for participation. PMID- 19682127 TI - Plasma carboxymethyl-lysine, an advanced glycation end product, and all-cause and cardiovascular disease mortality in older community-dwelling adults. AB - OBJECTIVES: To determine whether older adults with high plasma carboxymethyl lysine (CML), an advanced glycation end product, are at higher risk of all-cause and cardiovascular disease (CVD) mortality. DESIGN: Prospective cohort study. SETTING: Population-based sample of adults aged 65 and older residing in Tuscany, Italy. PARTICIPANTS: One thousand thirteen adults participating in the Invecchiare in Chianti study. MEASUREMENTS: Anthropometric measures, plasma CML, fasting plasma total, high-density and low-density lipoprotein cholesterol, triglycerides, glucose, creatinine. Clinical measures: medical assessment, diabetes mellitus, hypertension, coronary heart disease, heart failure, stroke, cancer. Vital status measures: death certificates and causes of death according to the International Classification of Diseases. Survival methods were used to examine the relationship between plasma CML and all-cause and CVD mortality, adjusting for potential confounders. RESULTS: During 6 years of follow-up, 227 (22.4%) adults died, of whom 105 died with CVD. Adults with plasma CML in the highest tertile had greater all-cause (hazard ratio (HR)=1.84, 95% confidence interval) CI)=1.30-2.60, P<.001) and CVD (HR=2.11, 95% CI=1.27-3.49, P=.003) mortality than those in the lower two tertiles after adjusting for potential confounders. In adults without diabetes mellitus, those with plasma CML in the highest tertile had greater all-cause (HR=1.68, 95% CI=1.15-2.44, P=.006) and CVD (HR=1.74, 95% CI=1.00-3.01, P=.05) mortality than those in the lower two tertiles after adjusting for potential confounders. CONCLUSION: Older adults with high plasma CML are at higher risk of all-cause and CVD mortality. PMID- 19682129 TI - Association between cognitive function and social support with glycemic control in adults with diabetes mellitus. AB - OBJECTIVES: To examine whether cognitive impairment in adults with diabetes mellitus is associated with worse glycemic control and to assess whether level of social support for diabetes mellitus care modifies this relationship. DESIGN: Cross-sectional analysis. SETTING: The 2003 Health and Retirement Study (HRS) Mail Survey on Diabetes and the 2004 wave of the HRS. PARTICIPANTS: Adults aged 50 and older with diabetes mellitus in the United States (N=1,097, mean age 69.2). MEASUREMENTS: Glycosylated hemoglobin (HbA1c) level; cognitive function, measured with the 35-point HRS cognitive scale (HRS-cog); sociodemographic variables; duration of diabetes mellitus; depressed mood; social support for diabetes mellitus care; self-reported knowledge of diabetes mellitus; treatments for diabetes mellitus; components of the Total Illness Burden Index related to diabetes mellitus; and functional limitations. RESULTS: In an ordered logistic regression model for the three ordinal levels of HbA1c (<7.0, 7.0-7.9, >or=8.0 mg/dL), respondents with HRS-cog scores in the lowest quartile had significantly higher HbA1c levels than those in the highest cognitive quartile (adjusted odds ratio=1.80, 95% confidence interval=1.11-2.92). A high level of social support for diabetes mellitus care modified this association; for respondents in the lowest cognitive quartile, those with high levels of support had significantly lower odds of having higher HbA1c than those with low levels of support (1.11 vs 2.87, P=.02). CONCLUSION: Although cognitive impairment was associated with worse glycemic control, higher levels of social support for diabetes mellitus care ameliorated this negative relationship. Identifying the level of social support available to cognitively impaired adults with diabetes mellitus may help to target interventions for better glycemic control. PMID- 19682130 TI - Mortality risk associated with physical and verbal abuse in women aged 50 to 79. AB - OBJECTIVES: To investigate whether midlife and older women who reported prior year physical abuse, verbal abuse, or both abuse types had higher mortality risk than peers who did not report prior-year abuse. DESIGN: Retrospective analysis. SETTING: Community. PARTICIPANTS: One hundred sixty-thousand six hundred seventy six community-dwelling women ages 50 to 79 at baseline enrolled in one of two major Women's Health Initiative (WHI) study components who responded to baseline abuse questions. Observational study enrollment was N=93,676 (1994-1998; 90 months average follow-up). Clinical trial enrollment was N=68,132 (1993-1998; 96 months average follow-up). MEASUREMENTS: Total mortality was measured from 1993 to 2005 using all available data sources. Blinded physician adjudicators measured cause-specific mortality. Ninety-six percent of death records were adjudicated. RESULTS: Prior-year self-reported abuse prevalence was 11.3%. Women who reported physical abuse had the highest age-adjusted mortality rate, followed by women who reported both abuse types. Abuse independently predicted mortality risk after controlling for age, education, ethnicity, and WHI component. High mortality risk remained for physically abused women (hazard ratio (HR)=1.54, 95% confidence interval (CI)=1.09-2.18) after adjusting for demographic and health-related factors. Further adjustment for psychosocial variables diminished this association (HR=1.40, 95% CI=0.93-2.11), but high risk remained. CONCLUSION: Community-dwelling middle-aged and older women who reported prior-year physical, verbal, or both types of abuse had significantly higher adjusted mortality risk than women who did not report abuse. These findings highlight the need for longitudinal research into prevention of abuse in later life and accompanying excess mortality and emphasize the importance of abuse prevention in later life. PMID- 19682131 TI - Clinical features, treatment practices, and hospital and long-term outcomes of older patients hospitalized with decompensated heart failure: The Worcester Heart Failure Study. AB - OBJECTIVES: To examine age-specific differences in clinical presentation, receipt of therapeutic practices and lifestyle recommendations, and hospital and long term survival in patients hospitalized for acute heart failure HF. DESIGN: Population-based study. SETTING: The Worcester Heart Failure Study, a population based study of residents of the a large Central New England metropolitan area hospitalized for decompensated HF at 11 greater-Worcester medical centers. PARTICIPANTS: Four thousand five hundred thirty-four patients hospitalized for decompensated HF during 1995 and 2000. MEASUREMENTS: Medical records were reviewed for demographic, clinical, and treatment characteristics and hospital survival status. Long-term follow-up of discharged hospital patients was conducted through 2005. Patients were compared according to four age groups (<65, 65-74, 75-84, and > or =85). RESULTS: Mean age was 76; 24.0% were aged 85 and older. Patients aged 75 and older were more likely to be female and to have multiple comorbidities, a lower body mass index at the time of hospitalization, and higher ejection fraction findings. Older patients were significantly more likely to receive symptom-modifying medications and less likely to receive disease-modifying medications than younger patients. Older age was directly associated with higher in-hospital, 30-day, and 1-year death rates in crude and multivariable-adjusted analyses. CONCLUSION: The results of this community-wide study suggest that clinical, treatment, and prognostic factors differ according to age in patients hospitalized for decompensated HF. These high-risk patients warrant special attention in future studies to improve their management and long term survival. PMID- 19682132 TI - Use of Medicare and Department of Veterans Affairs health care by veterans with dementia: a longitudinal analysis. AB - The objectives of this study were to examine longitudinal patterns of Department of Veterans Affairs (VA)-only use, dual VA and Medicare use, and Medicare-only use by veterans with dementia. Data on VA and Medicare use were obtained from VA administrative datasets and Medicare claims (1998-2001) for 2,137 male veterans who, in 1997, used some VA services, had a formal diagnosis of Alzheimer's disease or vascular dementia in the VA, and were aged 65 and older. Generalized ordered logit models were used to estimate the effects of patient characteristics on use group over time. In 1998, 41.7% of the sample were VA-only users, 55.4% were dual users, and 2.9% were Medicare-only users. By 2001, 30.4% were VA-only users, 51.5% were dual users, and 18.1% were Medicare-only users. Multivariate results show that greater likelihood of Medicare use was associated with older age, being white, being married, having higher education, having private insurance or Medicaid, having low VA priority level, and living in a nursing home or dying during the year. Higher comorbidities were associated with greater likelihood of dual use as opposed to any single system use. Alternatively, number of functional limitations was associated with greater likelihood of Medicare-only use and less likelihood of VA-only use. These results imply that different aspects of veterans' needs have differential effects on where they seek care. Efforts to coordinate care between VA and Medicare providers are necessary to ensure that patients receive high-quality care, especially patients with multiple comorbidities. PMID- 19682133 TI - Anemia and 9-year domain-specific cognitive decline in community-dwelling older women: The Women's Health and Aging Study II. AB - OBJECTIVES: To test the hypothesis that anemia (hemoglobin <12 g/dL) is associated with a faster rate of cognitive decline over 9 years in a community dwelling sample of women aged 70 to 80 at baseline. DESIGN: A population-based, prospective cohort study. SETTING: East Baltimore, Maryland. PARTICIPANTS: Four hundred thirty-six women sampled to be representative of the two-thirds least disabled women aged 70 to 80 at baseline (1994-1996). MEASUREMENTS: Nine-year trajectories of cognitive decline, analyzed using linear random effects models, in the domains of immediate verbal recall, delayed verbal recall, psychomotor speed, and executive function. RESULTS: At baseline and after adjustment for demographic and disease covariates, women with anemia were slower to complete a test of executive function; the difference in baseline function between women with anemia and those without was -0.43 standard deviations (SDs) (95% confidence interval (CI)=-0.74 to -0.13) on the Trail Making Test Part B. During follow-up, anemia was associated with a faster rate of decline in memory. Between baseline and Year 3, the difference in the rates of decline between women with anemia and those without was -0.18 SDs per year (95% CI=-0.29 to -0.06) on the Hopkins Verbal Learning Test (HVLT) and -0.15 SDs per year (95% CI=-0.26 to -0.04) on the HVLT-Delayed. CONCLUSION: Anemia was associated with poorer baseline performance on a test of executive function and with faster rates of decline on tests of immediate and delayed verbal recall. If this relationship is causal, it is possible that treatment of anemia could prevent or postpone cognitive decline. PMID- 19682134 TI - Description and students' perceptions of a required geriatric clerkship in postacute rehabilitative care. AB - This article describes medical students' evaluation of a geriatric clerkship in postacute rehabilitative care settings. This was a cross-sectional study of fourth-year medical students who completed a mandatory 2-week rotation at a postacute care facility. Students were provided with three instructional methods: Web-based interactive learning modules; small-group sessions with geriatric faculty; and Geriatric Interdisciplinary Care Summary (GICS), a grid that students used to formulate comprehensive interdisciplinary care plans for their own patients. After the rotation, students evaluated the overall clerkship, patient care activities, and usefulness of the three instructional methods using a 5-point Likert scale (1=poor to 5=excellent) and listed their area of future specialty. Of 156 students who completed the rotation, 117 (75%) completed the evaluation. Thirty (26%) chose specialties providing chronic disease management such as family, internal medicine, and psychiatry; 34 (29%) chose specialties providing primarily procedural services such as surgery, radiology, anesthesiology, pathology, and radiation oncology. Students rated the usefulness of the GICS as good to very good (mean+/-standard deviation 3.3+/-1.0). Similarly, they rated overall clerkship as good to excellent (3.8+/-1.0). Analysis of variance revealed no significant group difference in any of the responses from students with the overall clerkship (F(112, 4)=1.7, P=.20). Students rated the geriatric clerkship favorably and found the multimodal instruction to be useful. Even for students whose career choice was not primary care, geriatrics was a good model for interdisciplinary care training and could serve as a model for other disciplines. PMID- 19682135 TI - The epidemiology of dependence in older people in Nigeria: prevalence, determinants, informal care, and health service utilization. A 10/66 dementia research group cross-sectional survey. AB - OBJECTIVES: To describe the prevalence and determinants of dependence in older Nigerians and associations with informal care and health service utilization. DESIGN: A single-phase cross-sectional catchment area survey. SETTING: Dunukofia, a rural community in southeastern Nigeria. PARTICIPANTS: One thousand two hundred thirty-eight adults aged 65 and older, for whom full data were available on 914. MEASUREMENTS: The full 10/66 Dementia Research Group survey protocol was applied, including ascertainment of depression, cognitive impairment, physical impairments, and self-reported diagnoses. The interviewer rated dependence as not needing care, needing some care, or needing much care. The prevalence of dependence and the independent contribution of underlying health conditions were estimated. Sources of income, care arrangements, caregiver strain, and health service use are described according to level of dependence. RESULTS: The prevalence of dependence was 24.3% (95% confidence interval=22.1-26.5%), with a concentration in participants aged 80 and older. Only 1% of participants received a pension, and fewer than 7% had paid work. Those who were dependent were less likely than others to receive income from their family. Cognitive impairment, physical impairments, stroke, and depression were each independently associated with dependence. Depression made the largest contribution. Dependence was strongly associated with health service use (particularly private doctor and traditional healer services) and with high levels of out-of-pocket expenditure. CONCLUSION: In Nigeria, dependence is an important outcome given rapid demographic aging and increases in chronic disease prevalence in all developing regions. Enhancing the social protection of dependent older adults should be a policy priority. Cognitive and mental disorders are important contributors to disability and dependence; more attention should be given to their prevention, detection, and treatment. PMID- 19682136 TI - Black and white adult family members' attitudes toward a dementia diagnosis. AB - OBJECTIVES: To examine potential benefits of and barriers to diagnosis from the perspective of black and white adults directly affected by Alzheimer's disease (AD). DESIGN: Telephone survey. SETTING: Convenience sample recruited from two U.S. metropolitan areas. PARTICIPANTS: One hundred seventy-eight family members of people with AD, including current and former AD caregivers and immediate blood relatives of someone with AD. MEASUREMENTS: Respondents were asked to rate the importance of eight benefits of and 16 barriers to obtaining a diagnosis. RESULTS: Family members strongly endorse several benefits of obtaining a diagnosis, including getting information, finding out what is wrong with their relative, and prompting future planning. A majority of survey respondents did not endorse any barriers examined. Lack of a cure for AD and the belief that little can be done for someone with AD were the most frequently endorsed barriers. Black respondents endorsed five of the eight benefits more frequently than white respondents. CONCLUSION: Black and white adults with a family member who has received an diagnosis of AD perceive a range of benefits and few barriers to the diagnostic process examined in this study. Their positive experiences might be instructive to families considering pursuing a diagnosis and to physicians who may be reluctant to offer screening or referral because of the belief that families have little to gain. PMID- 19682137 TI - Leptin increases growth of primary ossification centers in fetal mice. AB - The effect of peripheral leptin on fetal primary ossification centers during the early phases of bone histogenesis was investigated by administration of leptin to pregnant mice. Fourteen pregnant mice were divided into two groups. The treated pregnant group was subcutaneously injected in the intrascapular region with supraphysiologic doses (2 mg kg(-1)) of leptin (Vinci Biochem, Firenze, Italy) in a volume of 0.1 mL per 10 g body weight, at the 7th, 9th and 11th day of gestation. The control group was treated with physiological solution in the same manner and same times as the treated group. The new-born mice were killed 1 day after birth and the primary ossification centers were stained with Alizarin Red S after diaphanizing the soft tissues in 1% potassium hydroxide. The development of both endochondral and intramembranous ossification centers was morphometrically analysed in long bones. The results showed that the ossification centers of mice born by mothers treated with leptin grow more rapidly in both length and cross sectional area compared with mice born by the untreated mothers. As the development of long bones depends on endochondral ossification occurring at proximal and distal epiphyseal plates as well as on intramembranous ossification along the periosteal surface, it appears that leptin activates the differentiation and proliferation of both chondrocytes and osteoblasts. The role of leptin as a growth factor of cartilage and bone is discussed in the light of the data reported in the literature. PMID- 19682139 TI - Conception date affects litter type and foetal sex ratio in female moose in Estonia. AB - 1. The Trivers-Willard model of optimal sex ratios predicts that in polygynous species mothers in better condition should produce more male than female offspring. However, empirical support for this hypothesis in mammals and especially ungulates has been equivocal. This may be because the fitness of mothers has been defined in different ways, reflecting morphological, physiological or behavioural measures of condition. In addition, factors other than maternal condition can influence a mother's fitness. Given that recent studies of wild ungulates have demonstrated the importance of the timing of conception and birth on offspring fitness, litters conceived at different stages of the rut might be expected to exhibit differences in types and embryonic sex ratio. 2. Based on a 6-year survey of the reproductive tracts of female moose harvested in Estonia, we investigated the effect of conception date on the types of litters produced and on the foetal sex ratio. 3. There was a clear relationship between conception date and litter characteristics. Overall, earlier conceived litters were more likely than those conceived late to contain multiple embryos and a high proportion of males. However, while foetal sex ratio varied nonlinearly with conception date in yearlings and subadults, no relationship was found in adults. 4. We conclude that female moose adjust foetal sex ratio and litter type/size depending on their age and the date of conception, and that these adjustments are in accordance with the Trivers-Willard hypothesis if females that conceive earlier are in better condition. PMID- 19682138 TI - The efficacy of end-to-end and end-to-side nerve repair (neurorrhaphy) in the rat brachial plexus. AB - Proximal nerve injury often requires nerve transfer to restore function. Here we evaluated the efficacy of end-to-end and end-to-side neurorrhaphy of rat musculocutaneous nerve, the recipient, to ulnar nerve, the donor. The donor was transected for end-to-end, while an epineurial window was exposed for end-to-side neurorrhaphy. Retrograde tracing showed that 70% donor motor and sensory neurons grew into the recipient 3 months following end-to-end neurorrhaphy compared to 40 50% at 6 months following end-to-side neurorrhaphy. In end-to-end neurorrhaphy, regenerating axons appeared as thick fibers which regained diameters comparable to those of controls in 3-4 months. However, end-to-side neurorrhaphy induced slow sprouting fibers of mostly thin collaterals that barely approached control diameters by 6 months. The motor end plates regained their control density at 4 months following end-to-end but remained low 6 months following end-to-side neurorrhaphy. The short-latency compound muscle action potential, typical of that of control, was readily restored following end-to-end neurorrhaphy. End-to-side neurorrhaphy had low amplitude and wide-ranging latency at 4 months and failed to regain control sizes by 6 months. Grooming test recovered successfully at 3 and 6 months following end-to-end and end-to-side neurorrhaphy, respectively, suggesting that powerful muscle was not required. In short, both neurorrhaphies resulted in functional recovery but end-to-end neurorrhaphy was quicker and better, albeit at the expense of donor function. End-to-side neurorrhaphy supplemented with factors to overcome the slow collateral sprouting and weak motor recovery may warrant further exploration. PMID- 19682140 TI - Great tits lay increasingly smaller clutches than selected for: a study of climate- and density-related changes in reproductive traits. AB - 1. The phenology of temperate environments and therefore timing of breeding has advanced in a number of bird species due to climate warming. Few studies, however, have examined the mechanisms behind the observed changes, the role of natural selection in them or the determinants of the selection. In other traits such as clutch size, even changes over years have been rarely studied. 2. We studied patterns and trends in timing of breeding, clutch size and fledgling production in the great tit Parus major in South-West Finland during 1953-2008, as well as natural selection on the timing and clutch size, based on fledgling production. We also examined connections between these parameters and a number of climatic and population intrinsic factors. 3. Laying date was earlier when the pre-breeding period was warm and tended to be earlier when breeding density was high, but it did not show any temporal change during the study period despite temporal increases in both explanatory factors. Number of fledglings decreased through declines in both mean clutch size and fledging success. Fledging success was better with higher breeding-time temperature and larger clutch size. Both the clutch size and fledging success were lower at higher breeding density. 4. Selection on laying date did not change through time, but there was a selection for early laying with high breeding-time temperature and high breeding density. Interestingly, in contrast to the decrease in reproductive output, the selection for larger than average clutch size strengthened with time, which was not explained by any tested factor. 5. We suggest that increasingly favourable conditions in winters have enhanced the survival and resulted in the observed increase in great tit breeding density. This may have most concerned young and otherwise low-quality individuals, which also most likely end up breeding in the increasingly occupied low-quality territories. This hypothesis was indicatively supported by increased within-year variation in both laying date and clutch size. The changes could also explain the lack of advancement in laying date as well as the increasing selection for large clutch sizes as the fittest individuals most likely occupy the best territories and lay largest clutches. PMID- 19682141 TI - Transgressive aggression in Sceloporus hybrids confers fitness through advantages in male agonistic encounters. AB - 1. We investigated agonistic behaviour and associated characteristics of Sceloporus woodi (Florida scrub lizard), Sceloporus undulatus (Eastern fence lizard) and their hybrids using staged territorial encounters. 2. These Sceloporus hybrids exhibit transgressive aggression and transgressive head-girth relative to the parental species and the transgressive aggression was specifically associated with an advantage in agonistic encounters. Our results suggest a hybrid advantage in natural habitats when defending and invading territories against either parental species. 3. We further analysed general advantages in agonistic encounters across the entire three-group system to elucidate characteristics that may be advantageous under specific circumstances. Individuals with larger body size (SVL) and greater aggression had an overall advantage in agonistic encounters; however, smaller individuals could win when slightly more aggressive and fatter, and less aggressive individuals could win when slightly larger, especially with greater head-girth. 4. The extreme hybrid phenotypes likely occurred through transgressive segregation, which has been implicated as a process through which homoploid, hybrid speciation can occur. Some form of ecological divergence is necessary, however, to impede parental gene flow. Our data suggest that ecological divergence could manifest in territorial species through transgressive aggression. PMID- 19682142 TI - Invasion success depends on invader body size in a size-structured mixed predation-competition community. AB - 1. The size of an individual is an important determinant of its trophic position and the type of interactions it engages in with other heterospecific and conspecific individuals. Consequently an individual's ecological role in a community changes with its body size over ontogeny, leading to that trophic interactions between individuals are a size-dependent and ontogenetically variable mixture of competition and predation. 2. Because differently sized individuals thus experience different biotic environments, invasion success may be determined by the body size of the invaders. Invasion outcome may also depend on the productivity of the system as productivity influences the biotic environment. 3. In a laboratory experiment with two poeciliid fishes the body size of the invading individuals and the daily amount of food supplied were manipulated. 4. Large invaders established persistent populations and drove the resident population to extinction in 10 out of 12 cases, while small invaders failed in 10 out of 12 trials. Stable coexistence was virtually absent. Invasion outcome was independent of productivity. 5. Further analyses suggest that small invaders experienced a competitive recruitment bottleneck imposed on them by the resident population. In contrast, large invaders preyed on the juveniles of the resident population. This predation allowed the large invaders to establish successfully by decreasing the resident population densities and thus breaking the bottleneck. 6. The results strongly suggest that the size distribution of invaders affects their ability to invade, an implication so far neglected in life history omnivory systems. The findings are further in agreement with predictions of life-history omnivory theory, that size-structured interactions demote coexistence along a productivity gradient. PMID- 19682145 TI - Risk of permanent brain injury during syncope. PMID- 19682143 TI - Do muscle mass, muscle density, strength, and physical function similarly influence risk of hospitalization in older adults? AB - OBJECTIVES: To examine the association between strength, function, lean mass, muscle density, and risk of hospitalization. DESIGN: Prospective cohort study. SETTING: Two U.S. clinical centers. PARTICIPANTS: Adults aged 70 to 80 (N=3,011) from the Health, Aging and Body Composition Study. MEASUREMENTS: Measurements were of grip strength, knee extension strength, lean mass, walking speed, and chair stand pace. Thigh computed tomography scans assessed muscle area and density (a proxy for muscle fat infiltration). Hospitalizations were confirmed by local review of medical records. Negative binomial regression models estimated incident rate ratios (IRRs) of hospitalization for race- and sex-specific quartiles of each muscle and function parameter separately. Multivariate models adjusted for age, body mass index, health status, and coexisting medical conditions. RESULTS: During an average 4.7 years of follow-up, 1,678 (55.7%) participants experienced one or more hospitalizations. Participants in the lowest quartile of muscle density were more likely to be subsequently hospitalized (multivariate IRR=1.47, 95% confidence interval (CI)=1.24-1.73) than those in the highest quartile. Similarly, participants with the weakest grip strength were at greater risk of hospitalization (multivariate IRR=1.52, 95% CI=1.30-1.78, Q1 vs. Q4). Comparable results were seen for knee strength, walking pace, and chair stands pace. Lean mass and muscle area were not associated with risk of hospitalization. CONCLUSION: Weak strength, poor function, and low muscle density, but not muscle size or lean mass, were associated with greater risk of hospitalization. Interventions to reduce the disease burden associated with sarcopenia should focus on increasing muscle strength and improving physical function rather than simply increasing lean mass. PMID- 19682147 TI - Novel interdisciplinary analgesic program. PMID- 19682148 TI - Prevalence of subclinical hypothyroidism in older patients with diabetes mellitus with poorly controlled dyslipidemia in China. PMID- 19682149 TI - Open mesh-plug inguinal hernia repair in the oldest old. PMID- 19682150 TI - Teaching in geriatric medicine in Cluj Napoca, Romania. PMID- 19682151 TI - Summer preference in the occurrence of takotsubo cardiomyopathy is independent of age. PMID- 19682152 TI - Day-of-week distribution of fatal and nonfatal ischemic stroke in elderly subjects. PMID- 19682153 TI - Long-term use of proton pump inhibitors: are they really safe? A case of delayed acute interstitial nephritis. PMID- 19682154 TI - Herpes simplex encephalitis with normal initial cerebrospinal fluid examination. PMID- 19682155 TI - How to manage overactive bladder in elderly individuals with dementia? A combined use of donepezil, a central acetylcholinesterase inhibitor, and propiverine, a peripheral muscarine receptor antagonist. PMID- 19682156 TI - Glucose impairment in older adults with diabetes mellitus and non-ST elevation acute myocardial infarction is linked to myocardial necrosis and systolic dysfunction. PMID- 19682157 TI - Acrocyanosis as a cause of pseudohypoglycemia. PMID- 19682159 TI - Six degrees of Apodemus separation. AB - Social network analysis has been widely used to help understand the transmission of human diseases. Its application to wildlife disease is very much in its infancy, largely because of the difficulty of recording contacts between wild animals. Sarah Perkins et al. have constructed contact networks for yellow-necked mice (Apodemus flavicollis) in the Italian Alps, comparing networks derived from radio-tracking and mark-recapture data. They found that the method producing the most informative data depended on population density. However, all networks had aggregated contact distributions, which is important for understanding disease transmission. PMID- 19682160 TI - Effect of non-random dispersal strategies on spatial coexistence mechanisms. AB - 1. Random dispersal leads to spatial coexistence via two mechanisms (emigration mediated and source-sink), both of which involve the movement of organisms from areas of higher to lower fitness. What is not known is whether such coexistence would occur if organisms dispersed non-randomly, using cues such as density and habitat quality to gauge fitness differences between habitats. Here, I conduct a comparative analysis of random and non-random dispersal strategies in a foodweb with a basal resource, top predator, and two intermediate consumers that exhibit a trade-off between competitive ability and predator susceptibility. 2. I find a striking contrast between density- and habitat-dependent dispersal in their effects on spatial coexistence. Dispersal in response to competitor and predator density facilitates coexistence while dispersal in response to habitat quality (resource productivity and predator pressure) inhibits it. Moreover, density dependent dispersal changes species' distribution patterns from interspecific segregation to interspecific aggregation, while habitat-dependent dispersal preserves the interspecific segregation observed in the absence of dispersal. Under density-dependent dispersal, widespread spatial coexistence results in an overall decline in the abundance of the inferior competitor that is less susceptible to predation and an overall increase in the abundance of the superior competitor that is more susceptible to predation. Under habitat-dependent dispersal, restricted spatial coexistence results in species' abundances being essentially unchanged from those observed in the absence of dispersal. 3. A key outcome is that when the superior competitor moves in the direction of increasing fitness but the inferior competitor does not, spatial coexistence is possible in both resource-poor and resource-rich habitats. However, when the inferior competitor moves in the direction of increasing fitness but the superior competitor does not, spatial coexistence is precluded in resource-poor habitats and greatly reduced in resource-rich habitats. This suggests that species specific differences may play an important role in driving spatial coexistence patterns. 4. The comparative framework yields predictions that can be tested with experiments that manipulate the relative mobilities of interacting species, or observational data on relative abundances and distribution patterns. PMID- 19682161 TI - Late surgical mitral valve repair after percutaneous repair with the MitraClip system. AB - Percutaneous approaches for treating mitral regurgitation are under investigation, including repair with the MitraClip percutaneous mitral repair system (Evalve, Inc., Menlo Park, CA, USA), which has undergone extensive preclinical and clinical evaluation in the EVEREST I and II trials. The procedure involves the transcatheter placement of one or two MitraClip devices under echocardiographic and fluoroscopic guidance to restore leaflet coaptation. A desirable feature of any percutaneous mitral valve (MV) repair system is that the device should not impede subsequent surgical repair if needed. To date, the majority of reported MV surgeries after MitraClip device implantation have occurred earlier, within one year of treatment. We herein describe four previously unreported cases of successful surgical MV repair up to five years after MitraClip device implantation, demonstrating that late MV repair remains possible, including after implantation of two clips. PMID- 19682162 TI - RhoA/ROK pathway related to the mechanism of higher susceptibility to spasm in RA than in IMA. AB - BACKGROUND AND AIM OF THE STUDY: By investigating the expression and function of RhoA/Rho kinase pathway in the radial artery (RA), internal mammary artery (IMA), and great saphenous vein (GSV), this study aimed to elucidate the mechanism for a higher susceptibility of spasm in the RA and provide an effective drug candidate to prevent and treat RA spasm. METHODS: RA, IMA, and GSV that would otherwise have been discarded were collected from 25 patients who underwent coronary artery bypass grafting. Eleven matched rings of RA, IMA, and GSV were used to evaluate the vasodilatory properties of 10(-7-)10(-4) mol/l of fasudil, a Rho-kinase inhibitor, by using in vitro organ chambers. Another 14 matched RA, IMA, and GSV were used to demonstrate the immunohistochemistry (IHC) of RhoA and mRNA of RhoA and Rho kinase. RESULTS: The maximal vasodilation of RA to fasudil was significantly greater than IMA. RhoA protein IHC staining was different in IMA, RA, and GSV (RA > GSV >IMA). The expression of RhoA and Rho kinase mRNA in the RA was significantly greater than in the IMA. CONCLUSIONS: The expression of RhoA/Rho kinase mRNA and protein and function in the RA were significantly stronger than in the IMA, suggesting that RhoA/Rho kinase pathway may be one mechanism by which RA is more susceptible to spasm than IMA. Rho kinase inhibitors can be effective drug candidates to prevent and treat vasospasm. PMID- 19682163 TI - Utility of cardiac CT and MRI for the diagnosis and preoperative assessment of cardiac paraganglioma. AB - BACKGROUND: Cardiac paragangliomas are rare cardiac tumors that are usually benign. Surgical excision can be curative. METHODS: We report a case of 39-year old male who, during the work up of acute coronary syndrome with coronary angiography, cardiac computed tomography (CT) and magnetic resonance imaging (MRI), was found to have cardiac paraganglioma. RESULTS: The tumor was intrapericardial, arising at the level of proximal left anterior descending artery. The tumor was completely resected and the postoperative course was uneventful. At 3-months follow-up the patient was asymptomatic with normal ventricular function. CONCLUSION: Cardiac CT and MRI are valuable in characterizing and preoperative planning of primary cardiac paragangliomas. PMID- 19682164 TI - Unusual modification of the cabrol shunt for control of hemorrhage in acute type a aortic dissection. AB - A patient with acute Type A dissection required complex root repair with composite graft. She developed life-threatening hemorrhage at the root of the aorta, which could not be controlled with usual measures. A modification of the original Cabrol shunt allowed for successful control of bleeding. PMID- 19682166 TI - A novel mechanism of failure to detect atrial arrhythmias by pacemakers and implantable cardioverter defibrillators. AB - A 64-year-old man with complete heart block, status post-Medtronic dual chamber pacemaker insertion, failed ablation for atrial tachycardia at an outside institution. Despite persistent palpitations and known unsuccessful ablation, pacemaker interrogation revealed no evidence of atrial arrhythmias. At electrophysiology study, burst pacing from the high right atrium and distal coronary sinus at 370 ms revealed bidirectional 2:1 interatrial conduction block. Left atrial burst pacing at 260 ms induced an atrial tachycardia (cycle length 340 ms) with 2:1 left to right atrial block and right atrial activation at 680 ms. The tachycardia was localized to the lateral left atrial roof. A series of ablation lesions from left to right superior pulmonary vein terminated the tachycardia. Left to right interatrial conduction block is a mechanism for underdetection of atrial arrhythmias with implantable devices not previously described. As the extent of atrial ablation increases, the incidence of this mechanism of underdetection may increase. Though devices are often considered ideal for atrial arrhythmia detection and are used in multiple trials, detection failures can occur despite appropriate device function. This case underscores the need for electrocardiographic monitoring in addition to device-based electrogram monitoring. PMID- 19682165 TI - Remote monitoring of patients with an implanted device and patients' outcomes: the potential for "win-win" dynamics. PMID- 19682167 TI - Supraventricular tachycardia with varied p-wave morphologies-what is the mechanism? PMID- 19682168 TI - EP image. Focal atrial tachycardia arising from the right superior pulmonary vein with an epicardial connection to the left atrium following circumferential pulmonary vein isolation. PMID- 19682169 TI - Dynamic registration of preablation imaging with a catheter geometry to guide ablation in a Swine model: validation of image integration and assessment of catheter navigation accuracy. AB - BACKGROUND: Catheter ablation of atrial and ventricular tachyarrhythmia involves anatomically based cardiac ablation strategies. CT and MRI images provide the most detailed cardiac anatomy available. Integration of these images into a mapping system should produce detailed and accurate models suitable to guide ablation. OBJECTIVE: The purpose of this study was to validate and assess the accuracy of a novel CT and MRI image integration algorithm designed to facilitate catheter navigation and ablation. METHODS: Using a lateral thoracotomy, markers were sutured to the epicardial surface of each cardiac chamber in 12 swine. Detailed CT/MRI anatomy was imported into the mapping system. The CT/MRI image was then integrated with a detailed catheter geometry of the relevant chamber using a new image integration algorithm. The epicardial markers, identified from the CT/MRI images, were then displayed on the surface of the integrated image. Guided only by the integrated CT/MRI, a single RF lesion was directed at the corresponding endocardial site for each epicardial marker. At autopsy, the distance from the endocardial RF lesion to the target site was assessed. RESULTS: The mean position error (CT/MRI) for the left atrium was 2.5 +/- 2.4 mm/5.1 +/- 3.9 mm, for the right atrium 6.2 +/- 6.5 mm/4.3 +/- 2.2 mm, for the right ventricle 6.2 +/- 4.3 mm/6.6 +/- 5.3 mm, and for the left ventricle 4.7 +/- 3.4 mm/3.1 +/- 2.7 mm. There was no cardiac perforation or tamponade. CONCLUSION: CT and MRI images can be effectively utilized for catheter navigation when integrated into a mapping system. This novel registration module with dynamic registration provides effective guidance for ablation. PMID- 19682170 TI - Clinical value of noninducibility by high-dose isoproterenol versus rapid atrial pacing after catheter ablation of paroxysmal atrial fibrillation. AB - BACKGROUND: AF can be induced by RAP or ISO in >85% of patients with PAF. METHODS: ISO was administered in escalating doses of 5, 10, 15, and 20 microg/min in 112 patients (age = 56 +/- 13 years) with PAF before radiofrequency catheter ablation. AF was inducible in 97 of 112 patients (87%) at a mean dose of 15 +/- 5 microg/min. RAP induced AF in the remaining 14 of 15 patients. Antral pulmonary vein (PV) isolation (APVI) was followed by ablation of complex fractionated atrial electrograms (CFAEs) as necessary to terminate AF and render AF noninducible in response to ISO. RESULTS: AF terminated during APVI in 72 of 111 patients (65%) and after APVI plus ablation of CFAEs in 11 of 111 patients (10%). In the remaining 28 patients (25%), sinus rhythm was restored by transthoracic cardioversion. RAP was performed in the last 61 consecutive patients who were rendered noninducible by ISO. RAP initiated AF in 20 of 61 patients (33%) and atrial flutter in 6 patients (10%). No additional ablation was performed if AF was induced with RAP; however, atrial flutter was targeted. At 12 +/- 5 months, 63/75 patients (84%) who were noninducible by ISO and 2 of 8 (25%) who still were reinducible by ISO were free from recurrent AF after a single ablation procedure without antiarrhythmic drugs (P = 0.001). AF recurred in 20 of 36 patients (56%) who required cardioversion for persistent AF after ablation (P < 0.001). Among the 61 patients who also underwent RAP, 12 of 20 (60%) who were, and 31 of 41 (76%) who were not inducible by RAP were free from recurrent AF (P = 0.21). The accuracy of noninducibility as a predictor of clinical outcome was 83% with ISO and 64% by RAP (P = 0.03). CONCLUSIONS: The response to isoproterenol after catheter ablation of PAF more accurately predicts clinical outcome than the response to RAP. PMID- 19682171 TI - Focal ventricular tachycardia arising from the epicardial crux of the heart after a remote inferior myocardial infarction. PMID- 19682172 TI - Differential cytokine expression by human dendritic cells in response to different Porphyromonas gingivalis capsular serotypes. AB - AIM: Capsular polysaccharides play an important role in the virulence of Gram positive and Gram-negative bacteria. In Porphyromonas gingivalis, six serotypes have been described based on capsular antigenicity and its pathogenicity has been correlated both in vitro and in animal models. This study aimed to investigate the differential response of human dendritic cells (DCs) when stimulated with different P. gingivalis capsular serotypes. MATERIALS AND METHODS: Using different multiplicity of infection (MOI) of the encapsulated strains K1-K6 and the non-encapsulated K(-) strain of P. gingivalis, the mRNA expression levels for interleukin (IL)-1beta, IL-2, IL-5, IL-6, IL-10, IL-12, IL-13, interferon (IFN) gamma, tumour necrosis factor (TNF)-alpha, and TNF-beta in stimulated DCs were quantified by real-time reverse transcription-polymerase chain reaction. RESULTS: All P. gingivalis capsular serotypes induced a T-helper type 1 (Th1) pattern of cytokine expression. K1- and K2-stimulated DCs expressed higher levels of IL 1beta, IL-6, IL-12p35, IL-12p40, and IFN-gamma and at lower MOI than DCs stimulated with the other strains. CONCLUSIONS: These results demonstrate a differential potential of P. gingivalis capsular serotypes to induce DC responses and a higher capacity of strains K1 W83 and K2 HG184 than other K serotypes to trigger cytokine expression. PMID- 19682173 TI - The influence of a probiotic milk drink on the development of gingivitis: a pilot study. AB - AIM: The aim of this study was to determine the effect of a probiotic milk drink on gingival health and the development of experimental gingivitis. MATERIAL AND METHODS: Fifty volunteer students took part in a parallel-designed non-blinded study. The test group drank a probiotic drink once a day; the control group did not receive any product to drink. After 8 weeks, individual mechanical plaque control was interrupted for 96 h. Papilla bleeding index, interproximal plaque and Turesky plaque index (PI) were recorded at baseline, after 8 weeks and again 96 h later. At the same time points, gingival crevicular fluid had been collected for analysis of polymorphonuclear elastase, myeloperoxidase (MPO) and matrix metalloproteinase-3 (MMP-3). RESULTS: Interproximal PI and papillary bleeding were not different between the groups. In the test group, elastase activity and MMP-3 amount were significantly lower after the intake of the probiotic milk drink (p<0.001 and 0.016). There was a significant increase of MPO activity in the control group; both groups were different at the end of the study (p=0.014). CONCLUSIONS: The data suggest a beneficial effect of the probiotic milk drink on gingival inflammation. PMID- 19682175 TI - Sinonasal undifferentiated carcinoma metastatic to the skin. AB - Sinonasal undifferentiated carcinoma (SNUC) is an aggressive malignancy of disputed histogenesis that arises in the sinonasal tract and has an extremely poor prognosis. Despite multimodality treatment with surgical resection, radiation, and chemotherapy, recurrence is common. The tumor spreads by direct local extension, but also metastasizes to lymph nodes, brain, lung and bone. To date, reports of metastasis to the skin have not been published. We report a case of a 58-year-old woman diagnosed with SNUC who underwent surgical resection of the tumor followed by chemoradiation. The tumor soon recurred, and she required wide re-excision. Two months after this procedure, she developed multiple dermal nodules in the head and neck region, clinically suspicious for metastases. Biopsy of a nodule from the right neck revealed a poorly differentiated carcinoma, with morphological and immunohistochemical findings consistent with a metastasis of the patient's known SNUC. We conclude that the skin may be a rare site of metastasis of SNUC, and in some cases may be the presenting sign of tumor recurrence despite aggressive multimodality treatment. PMID- 19682176 TI - Role of the epidermal barrier in atopic dermatitis. AB - The skin's permeability barrier protects against extensive water loss and prevents the entry into the skin of harmful substances like irritants, allergens and microorganisms. The permeability barrier is mainly located in the stratum corneum and consists of corneocytes and a lipid-enriched intercellular domain. The barrier is formed during epidermal differentiation. In atopic dermatitis the skin barrier is disturbed already in non-lesional skin. The disturbed skin barrier allows the entry of environmental allergens from house dust mites, animal dander and grass pollen into the skin. In predisposed individuals these allergens may trigger via immunologic pathways the inflammation of atopy. The causes for the disturbed epidermal skin barrier are changes in skin lipids and in epidermal differentiation, in particular filaggrin mutations. Filaggrin mutations lead to a disturbed skin barrier and dry skin which are hallmarks in atopic dermatitis. Therapeutic agents influence the skin barrier differently; topical therapy with potent corticosteroids does not lead to the repair of the barrier in atopic dermatitis, whereas therapy with the calcineurin inhibitors and lipid-containing emulsions support barrier repair. PMID- 19682174 TI - Periodontal status of teeth with crown-root fractures: results two years after adhesive fragment reattachment. AB - AIM: This series of case reports evaluated the impact of adhesive crown-root fragment reattachment in periodontally healthy teeth suffering from crown-root fractures on various parameters of periodontal health over a time course of 2 years. MATERIAL AND METHODS: A total of 20 teeth with crown-root fractures in 18 periodontally healthy subjects were evaluated. After open-flap access, crown-root fragments were adhesively reattached to the root stub. In all cases, the vertical difference between the alveolar bone crest and the fracture line was or = 10 mmHg). The Risk Score for predicting significant portal hypertension was 14.2 - 7.1 x log(10) (platelet [10(9)/L]) + 4.2 x log(10) (bilirubin [mg/dL]). The area under the receiver operator curve (AUC) curve was 0.91 (95% confidence interval [CI], 0.84-0.98). The optimized cut-off value (Risk Score = -1.0) offered a sensitivity of 88% (95% CI, 62-98%) and a specificity of 86% (95% CI, 72-94%). The AUC of the Risk Score in predicting varices was 0.82 (95% CI, 0.67-0.98). The cut-off had a sensitivity of 82% (95% CI, 48-97%) and a specificity of 76% (95% CI, 62-86%). CONCLUSION: A predictive model that uses readily-available laboratory results may reliably identify advanced fibrosis patients with clinically-significant portal hypertension as well as esophageal varices. However, before accepted, the results of the current study certainly should be validated in larger prospective cohorts. PMID- 19682197 TI - Delayed endoscopy as a risk factor for in-hospital mortality in cirrhotic patients with acute variceal hemorrhage. AB - BACKGROUND AND AIMS: Risk factors for mortality in acute variceal hemorrhage remain incompletely understood. Whether endoscopy timing is associated with risk of mortality has not been investigated. We aimed to investigate risk factors for in-hospital mortality in cirrhotic patients with acute variceal hemorrhage, with emphasis on endoscopy timing. METHODS: Three hundred and eleven (73% male and 23% female) consecutive cirrhotic patients presenting with acute variceal hemorrhage from July 2004 to July 2007 were investigated. The univariate association of endoscopy timing as the predictor for in-hospital mortality was examined. Independent risk factors for mortality were determined by multivariate logistic regression analysis consisting of clinical, laboratory and endoscopic parameters. RESULTS: Twenty-five (8.04%) patients died within admission. By plotting the receiver operating curve of endoscopy timing for mortality, we selected 15 h as the optimal cut-off point to define delayed endoscopy. Multivariate regression analysis revealed that independent risk factors predictive for in-hospital mortality included delayed endoscopy performed 15 h after admission (adjusted odds ratio [aOR] = 3.67; 95% confidence interval [CI], 1.27-10.39), every point increment of model for end-stage liver disease (MELD) score (aOR = 1.16; 95% CI, 1.07-1.25), failure of the first endoscopy (aOR = 4.36; 95% CI, 1.54-12.30) and hematemesis as the chief complaint (compared with melena, aOR = 8.66; 95% CI, 1.06-70.94). CONCLUSION: Delayed endoscopy for more than 15 h, high MELD score, failure of the first endoscopy and hematemesis are independent risk factors for in-hospital mortality in cirrhotic patients with acute variceal hemorrhage. PMID- 19682198 TI - Education and imaging. Gastrointestinal: colon cancer with octopus suckers. PMID- 19682199 TI - Education and imaging. Gastrointestinal: gastric volvulus. PMID- 19682200 TI - Education and imaging. Gastrointestinal: virtual CT ileoscopy in terminal ileitis. PMID- 19682201 TI - Education and imaging. Hepatobiliary and pancreatic: torsion of an accessory spleen. PMID- 19682202 TI - Education and imaging. Hepatobiliary and pancreatic: fascioliasis. PMID- 19682205 TI - Involvement of glutamate in retinal protection against ischemia/reperfusion damage induced by post-conditioning. AB - Retinal ischemia could provoke blindness and there is no effective treatment against retinal ischemic damage. Brief intermittent ischemia applied during the onset of reperfusion (i.e., post-conditioning) protects the retina from ischemia/reperfusion injury. Multiple evidences support that glutamate is implicated in retinal ischemic damage. We investigated the involvement of glutamate clearance in post-conditioning-induced protection. For this purpose, ischemia was induced by increasing intra-ocular pressure for 40 min, and 5 min after reperfusion, animals underwent seven cycles of 1 min/1 min ischemia/reperfusion. One, three, or seven days after ischemia, animals were subjected to electroretinography and histological analysis. The functional and histological protection induced by post-conditioning was evident at 7 (but not 1 or 3) days post-ischemia. An increase in Muller cell glial fibrillary acidic protein (GFAP) levels was observed at 1, 3, and 7 days after ischemia, whereas post-conditioning reduced GFAP levels of Muller cells at 3 and 7 days post ischemia. Three days after ischemia, a significant decrease in glutamate uptake and glutamine synthetase activity was observed, whereas post-conditioning reversed the effect of ischemia. The intravitreal injection of supraphysiological levels of glutamate mimicked electroretinographic and histological alterations provoked by ischemia, which were abrogated by post-conditioning. These results support the involvement of glutamate in retinal protection against ischemia/reperfusion damage induced by post-conditioning. PMID- 19682204 TI - Docosahexaenoic acid promotes hippocampal neuronal development and synaptic function. AB - Docosahexaenoic acid (DHA, 22:6n-3), the major polyunsaturated fatty acid accumulated in the brain during development, has been implicated in learning and memory, but underlying cellular mechanisms are not clearly understood. Here, we demonstrate that DHA significantly affects hippocampal neuronal development and synaptic function in developing hippocampi. In embryonic neuronal cultures, DHA supplementation uniquely promoted neurite growth, synapsin puncta formation and synaptic protein expression, particularly synapsins and glutamate receptors. In DHA-supplemented neurons, spontaneous synaptic activity was significantly increased, mostly because of enhanced glutamatergic synaptic activity. Conversely, hippocampal neurons from DHA-depleted fetuses showed inhibited neurite growth and synaptogenesis. Furthermore, n-3 fatty acid deprivation during development resulted in marked decreases of synapsins and glutamate receptor subunits in the hippocampi of 18-day-old pups with concomitant impairment of long term potentiation, a cellular mechanism underlying learning and memory. While levels of synapsins and NMDA receptor subunit NR2A were decreased in most hippocampal regions, NR2A expression was particularly reduced in CA3, suggesting possible role of DHA in CA3-NMDA receptor-dependent learning and memory processes. The DHA-induced neurite growth, synaptogenesis, synapsin, and glutamate receptor expression, and glutamatergic synaptic function may represent important cellular aspects supporting the hippocampus-related cognitive function improved by DHA. PMID- 19682207 TI - The blood-brain barrier transport and cerebral distribution of guanidinoacetate in rats: involvement of creatine and taurine transporters. AB - Although the cerebral accumulation of guanidinoacetate (GAA) contributes to neurological complications in S-adenosylmethionine:guanidinoacetate N methyltransferase (GAMT) deficiency, how GAA is abnormally distributed in the brain remains unknown. The purpose of this study was to investigate the transport of GAA across the blood-brain barrier (BBB) and in brain parenchymal cells in rats. [(14)C]GAA microinjected into the rat cerebrum was not eliminated from the brain, implying the negligible contribution of GAA efflux transport across the BBB. In contrast, in vivo analysis and an uptake study by TR-BBB cells, a rat in vitro BBB model, revealed that GAA was transported from the circulating blood across the BBB most likely via a creatine transporter (CRT). Although CRT at the BBB is almost saturated by endogenous creatine under physiological conditions, the creatine level in the blood significantly decreases in GAMT deficiency. This might lead to the increase of CRT-mediated blood-to-brain transport of GAA at the BBB. Furthermore, [(14)C]GAA was taken up by brain parenchymal cells in a concentrative manner most likely via taurine transporter and CRT. These characteristics of GAA transport across the BBB and in the brain parenchymal cells could be the key factors that facilitate GAA accumulation in the brains of patients with GAMT deficiency. PMID- 19682206 TI - Astrocytes are poised for lactate trafficking and release from activated brain and for supply of glucose to neurons. AB - Brain is a highly-oxidative organ, but during activation, glycolytic flux is preferentially up-regulated even though oxygen supply is adequate. The biochemical and cellular basis of metabolic changes during brain activation and the fate of lactate produced within brain are important, unresolved issues central to understanding brain function, brain images, and spectroscopic data. Because in vivo brain imaging studies reveal rapid efflux of labeled glucose metabolites during activation, lactate trafficking among astrocytes and between astrocytes and neurons was examined after devising specific, real-time, sensitive enzymatic fluorescent assays to measure lactate and glucose levels in single cells in adult rat brain slices. Astrocytes have a 2- to 4-fold faster and higher capacity for lactate uptake from extracellular fluid and for lactate dispersal via the astrocytic syncytium compared to neuronal lactate uptake from extracellular fluid or shuttling of lactate to neurons from neighboring astrocytes. Astrocytes can also supply glucose to neurons as well as glucose can be taken up by neurons from extracellular fluid. Astrocytic networks can provide neuronal fuel and quickly remove lactate from activated glycolytic domains, and the lactate can be dispersed widely throughout the syncytium to endfeet along the vasculature for release to blood or other brain regions via perivascular fluid flow. PMID- 19682208 TI - Kaempferol protects against rat striatal degeneration induced by 3-nitropropionic acid. AB - 3-Nitropropionic acid (NPA) produces degeneration of striatum and some neurological disturbances characteristic of Huntington's disease in rodents and primates. We have shown that the flavonoid kaempferol largely reduced striatal damage induced by cerebral ischaemia-reperfusion in rats (Lopez-Sanchez et al. 2007). In this work, we report that intraperitoneal (i.p.) administration of kaempferol affords an efficient protection against NPA-induced neurodegeneration in Wistar rats. We studied the effects of daily i.p. injections of 7, 14 and 21 mg of kaempferol/kg body weight during the NPA-treatment (25 mg/kg body weight/12 h i.p., for 5 days) on the neurological deficits, degeneration of rat striatum and oxidative stress markers. Intraperitoneal injections of 14-21 mg of kaempferol/kg body weight largely attenuated motor deficit and delayed mortality. The higher dose of kaempferol prevented the appearance of NPA-induced striatal lesions up to the end of treatment, as revealed by haematoxylin-eosin and TUNEL staining, and also NPA-induced oxidative stress, because it blocked the fall of reduced glutathione and the increase of protein nitrotyrosines in NPA-treated rats. It was found that striatal degeneration was associated with calpains activation and a large inactivation of creatine kinase, which were also prevented when the higher doses of kaempferol were administered. PMID- 19682209 TI - The neurokinin-1 receptor modulates the methamphetamine-induced striatal apoptosis and nitric oxide formation in mice. AB - In a previous study we showed that pharmacological blockade of the neurokinin-1 receptors attenuated the methamphetamine (METH)-induced toxicity of the striatal dopamine terminals. In the present study we examined the role of the neurokinin-1 receptors on the METH-induced apoptosis of some striatal neurons. To that end, we administered a single injection of METH (30 mg/kg, i.p.) to male mice. METH induced the apoptosis (terminal deoxyncleotidyl transferase-mediated dUTP nick end labeling) of approximately 20% of striatal neurons. This percentage of METH induced apoptosis was significantly attenuated by either a single injection of the neurokinin-1 receptor antagonist, 17-beta-hydroxy-17-a-ethynyl-5-a androstano[3,2-beta]pyrimido[1,2-a]benzimidazole (WIN-51,708) (5 mg/kg, i.p.), or the ablation of the striatal interneurons expressing the neurokinin-1 receptors (cholinergic and somatostatin) with the selective neurotoxin [Sar(9),Met(O(2))(11)] substance P-saporin. Next we assessed the levels of striatal 3-nitrotyrosine (3-NT) by HPLC and immunohistochemistry. METH increased the levels of striatal 3-NT and this increase was attenuated by pre-treatment with WIN-51,708. Our data support the hypothesis that METH-induced striatal apoptosis occurs via a mechanism involving the neurokinin-1 receptors and the activation of nitric oxide synthesis. Our findings are relevant for the treatment of METH abuse and may be relevant to certain neurological disorders involving the dopaminergic circuitry of the basal ganglia. PMID- 19682211 TI - Cytoprotective effect of polypeptide from Chlamys farreri on neuroblastoma (SH SY5Y) cells following HO exposure involves scavenging ROS and inhibition JNK phosphorylation. AB - Oxidative stress has long been linked to cell death in many neurodegenerative conditions. Treatment with antioxidants is a promising approach for slowing disease progression. In this study, we used the neuroblastoma SH-SY5Y cells as an in vitro model to first assess the effect of polypeptide from Chlamys farreri (PCF), a natural marine antioxidant, on H(2)O(2)-induced neuronal cell death. Pre treatment of SH-SY5Y cells with PCF inhibited H(2)O(2)-induced cell death in a concentration-dependent manner. In parallel, intracellular reactive oxygen species generation and lipid peroxidation were inhibited by PCF. Under severe H(2)O(2) insult, PCF promoted endogenous antioxidant defense components including glutathione peroxidase, catalase, superoxide dismutase, and glutathione. PCF also protected DNA from oxidative damage and enhanced the removal of 8-oxo-7,8-dihydro 2'-deoxyguanosine from DNA. Further, we found that PCF potentially prevented H(2)O(2)-induced cell apoptosis. When investigated mitogen-activated protein kinase signaling pathway, we found that pre-treatment of cells with PCF significantly blocked H(2)O(2)-induced phosphorylation of c-Jun N-terminal kinase of the mitogen-activated protein kinase family. However, PCF had little inhibitory effect on the H(2)O(2)-induced activation of extracellular signal regulated kinase. Taken together, these data demonstrate that PCF prevents oxidative stress-induced reactive oxygen species production and c-Jun N-terminal kinase activation and may be useful in the treatment of neurodegenerative diseases. PMID- 19682212 TI - Aftercare of the complete denture patient. AB - Complete denture treatment does not end with the insertion of the finished prosthesis. Regular denture maintenance by the patient and periodic consultation with the prosthodontist are essential for the long-term success of treatment. This article attempts to focus on the points to be considered in the aftercare of the complete denture patient. PMID- 19682210 TI - Penelope's web: using alpha-latrotoxin to untangle the mysteries of exocytosis. AB - For more than three decades, the venom of the black widow spider and its principal active components, latrotoxins, have been used to induce release of neurotransmitters and hormones and to study the mechanisms of exocytosis. Given the complex nature of alpha--latrotoxin (alpha-LTX) actions, this research has been continuously overshadowed by many enigmas, misconceptions and perpetual changes of the underlying hypotheses. Some of the toxin's mechanisms of action are still not completely understood. Despite all these difficulties, the extensive work of several generations of neurobiologists has brought about a great deal of fascinating insights into pre-synaptic processes and has led to the discovery of several novel proteins and synaptic systems. For example, alpha-LTX studies have contributed to the widespread acceptance of the vesicular theory of transmitter release. Pre-synaptic receptors for alpha-LTX--neurexins, latrophilins and protein tyrosine phosphatase sigma--and their endogenous ligands have now become centrepieces of their own areas of research, with a potential of uncovering new mechanisms of synapse formation and regulation that may have medical implications. However, any future success of alpha-LTX research will require a better understanding of this unusual natural tool and a more precise dissection of its multiple mechanisms. PMID- 19682213 TI - In vitro tensile bond strength of denture repair acrylic resins to primed base metal alloys using two different processing techniques. AB - PURPOSE: Approximately 38% of removable partial denture (RPD) failures involve fracture at the alloy/acrylic interface. Autopolymerizing resin is commonly used to repair RPDs. Poor chemical bonding of repair acrylic to base metal alloys can lead to microleakage and failure of the bond. Therefore, ideal repair techniques should provide a strong, adhesive bond. This investigation compared the tensile bond strength between cobalt-chromium (Super Cast, Pentron Laboratory Technologies, Llc., Wallingford, CT) and nickel-chromium (Rexalloy, Pentron Laboratory Technologies, Llc.) alloys and autopolymerized acrylic resin (Dentsply Repair Material, Dentsply Int, Inc, York, Pa) using three primers containing different functional monomers [UBar (UB), Sun Medical Co., Ltd., Shiga, Japan: Alloy Primer (AP) Kuraray Medical Inc., Okayama, Japan; and MR Bond (MRB) Tokyuyama Dental Corp., Tokyo, Japan] and two processing techniques (bench cure and pressure-pot cure). MATERIAL AND METHODS: One hundred and twenty eight base metal alloy ingots were polished, air abraded, and ultrasonically cleaned. The control group was not primed. Specimens in the test groups were primed with one of the three metal primers. Autopolymerized acrylic resin material was bonded to the metal surfaces. Half the specimens were bench cured, and the other half were cured in a pressure pot. All specimens were stored in distilled water for 24 hours at 37 degrees C. The specimens were debonded under tension at a crosshead speed of 0.05 cm/min. The forces at which the bond failed were noted. Data were analyzed using ANOVA. Fisher's PLSD post hoc test was used to determine significant differences (p < 0.05). Failure modes of each specimen were evaluated under a dissecting microscope. RESULTS: Significant differences in bond strength were observed between combinations of primers, curing methods, and alloys. Primed sandblasted specimens that were pressure-pot-cured had significantly higher bond strengths than primed sandblasted bench-cured specimens. The pressure-pot-curing method had a significant effect on bond strength of all specimens except Co-Cr alloy primed with UB. The highest bond strength was observed for both Co-Cr and Ni-Cr alloys that were sandblasted, primed with MRB, and pressure-pot cured. Co Cr alloys primed with UB had the lowest bond strength whether bench cured or pressure-pot cured. Primed specimens generally experienced cohesive bond failures within the primer or acrylic resin. Nonprimed specimens generally experienced adhesive bond failures at the resin/metal interface. CONCLUSIONS: Within the limitations of this study, MRB provided the highest bond strength to both Ni-Cr and Co-Cr alloys. Generally, bond strength improved significantly when specimens were primed. Pressure-pot curing, in most cases, resulted in higher bond strength than bench curing. The results of this in vitro study suggest that MRB metal primer can be used to increase bond strength of autopolymerized repair acrylic resin to base metal alloys. Curing autopolymerized acrylic under pressure potentially increases bond strength. PMID- 19682214 TI - Fracture loads of all-ceramic crowns under wet and dry fatigue conditions. AB - PURPOSE: The aim of this study was to test the hypothesis that fracture loads of fatigued dental ceramic crowns are affected by testing environment and luting cement. MATERIALS AND METHODS: One hundred and eighty crowns were prepared from bovine teeth using a lathe. Ceramic crowns were prepared from three types of ceramic systems: an alumina-infiltrated ceramic, a lithia-disilicate-based glass ceramic, and a leucite-reinforced ceramic. For each ceramic system, 30 crowns were cemented with a composite resin cement, and the remaining 30 with a resin modified glass ionomer cement. For each ceramic system and cement, ten specimens were loaded to fracture without fatiguing. A second group (n = 10) was subjected to cyclic fatigue and fracture tested in a dry environment, and a third group (n = 10) was fatigued and fractured in distilled water. The results were statistically analyzed using one-way ANOVA and Tukey HSD test. RESULTS: The fracture loads of ceramic crowns decreased significantly after cyclic fatigue loading (p 0.05). CONCLUSIONS: Tribochemical silica coating and the application of dichloromethane after the silane coupling agent were effective surface preparations for the bonding of a highly filled gingival shade composite resin to a denture base resin, however, the bond durability of these treatments may be insufficient. PMID- 19682217 TI - Palatal anchorage for the retention of interim removable prostheses. AB - This paper describes a technique that involves the use of palatal implants to retain a maxillary interim prosthesis when extensive bone graft procedures are performed. The rationale is that some bone graft procedures require the removal of the denture flanges for graft success. Once the denture flanges are removed, the denture loses all its retention capabilities, making this lengthy interim phase difficult for the patient. While the use of palatal implants has been documented extensively, limited information is available to describe the use of palatal implants for prosthetic reasons. PMID- 19682218 TI - Tooth reduction guide using silicone registration material along with vacuum formed thermoplastic matrix. AB - Adequate tooth reduction is a prerequisite for function, esthetics, and longevity of fixed restorations. A tooth reduction guide may be useful for establishing the proper angulation of the tooth and maximizing periodontal health and restorative success. This article describes a simple and versatile technique for an accurate evaluation of tooth reduction for fixed restorations by using a color-contrasting positive guide of a silicone occlusal registration material processed inside a vacuum-formed clear thermoplastic matrix. PMID- 19682219 TI - Removable partial denture in combination with a milled fixed partial prosthesis as interim restorations in long-term treatment. AB - Interim restorations are frequently used in prosthodontic treatments. Many complex situations require the combination of fixed and removable partial prostheses. An appropriate interim restoration design that accurately implements the treatment plan is necessary to prepare the oral cavity for the prostheses, and to contribute to the preservation and health of remaining natural teeth, bone support, and gingival tissues. This report describes a modified technique for construction of interim restorations with a combination of fixed and removable partial prostheses. The technique consists of the construction of a milled fixed prosthesis and removable partial denture with metallic framework for use during extensive treatment, improving masticatory function and esthetics and preserving the periodontal health of supporting structures. This interim restoration can also serve as a template for the definitive restoration, allowing patient and dentist to evaluate appearance and function and helping to ensure the success of the definitive restoration. PMID- 19682220 TI - Comparison of the passivity between cast alloy and laser-welded titanium overdenture bars. AB - PURPOSE: The purpose of this study was to investigate the fit of cast alloy overdenture and laser-welded titanium-alloy bars by measuring induced strain upon tightening of the bars on a master cast as well as a function of screw tightening sequence. MATERIALS AND METHODS: Four implant analogs were secured into Type IV dental stone to simulate a mandibular edentulous patient cast, and two groups of four overdenture bars were fabricated. Group I was four cast alloy bars and Group II was four laser-welded titanium bars. The cast alloy bars included Au-Ag-Pd, Pd Ag-Au, Au-Ag-Cu-Pd, and Ag-Pd-Cu-Au, while the laser-welded bars were all Ti-Al-V alloy. Bars were made from the same master cast, were torqued into place, and the total strain in the bars was measured through five strain gauges bonded to the bar between the implants. Each bar was placed and torqued 27 times to 30 Ncm per screw using three tightening sequences. Data were processed through a strain amplifier and analyzed by computer using StrainSmart software. Data were analyzed by ANOVA and Tukey's post hoc test. RESULTS: Significant differences were found between alloy types. Laser-welded titanium bars tended to have lower strains than corresponding cast bars, although the Au-Ag-Pd bar was not significantly different. The magnitudes of total strain were the least when first tightening the ends of the bar. CONCLUSIONS: The passivity of implant overdenture bars was evaluated using total strain of the bar when tightening. Selecting a high modulus of elasticity cast alloy or use of laser-welded bar design resulted in the lowest average strain magnitudes. While the effect of screw tightening sequence was minimal, tightening the distal ends first demonstrated the lowest strain, and hence the best passivity. PMID- 19682221 TI - Force needed to separate acrylic resin from primed and unprimed frameworks of different designs. AB - PURPOSE: Poor mechanical and chemical bondings at the interface between a framework and denture base resin have been responsible for many removable partial denture failures. This study tested the force necessary to separate acrylic resin bases from test frameworks using different acrylic retention designs (smooth metal plate, metal plate with bead retention, lattice retention, and mesh retention). The force needed to separate acrylic resin from primed test frameworks was also measured. MATERIALS AND METHODS: Eighty chromium-cobalt test frameworks were fabricated using preformed wax patterns and cast according to manufacturer's instructions. Half the specimens were primed prior to acrylic processing. The same base acrylic was used for all specimens. Separation forces that fractured acrylic resin from test frameworks were generated by a universal testing machine at a crosshead speed of 25 mm/min. Loads at failure and types of failure were recorded. Data were analyzed using ANOVA. RESULTS: The mean separation force of acrylic resin from unprimed retention designs was highest for the metal plate with beads (3.1 kN), followed by mesh (2.8 kN) and lattice (2.1 kN), and lowest (0.1 kN) for the smooth metal plate. The mean separation force for primed acrylic retention designs was highest for the metal plate with beads (4.2 kN), followed by mesh (3.4 kN) and smooth metal plate (3.0 kN), and lowest for lattice retention (2.6 kN). Bond failure occurred both adhesively at the interface between metal and acrylic resin and cohesively within the acrylic resin. Cohesive bond failure increased when specimens were primed. The rate of cohesive bond failure remained the same for primed mesh retention specimens. CONCLUSIONS: Significantly increased force was necessary to separate the acrylic from each design of primed test specimens compared with unprimed specimens of the same design. The primed metal plate with beads exhibited significantly greater separation force than the other three designs. Primed mesh had significantly greater separation force values than primed lattice and smooth metal plate. Primed lattice was significantly less retentive than the other three primed designs. Except for the retentive mesh specimens, there was higher occurrence of cohesive failures in the acrylic resin when the frameworks were primed. PMID- 19682222 TI - Effect of light-emitting diode (LED) curing modes on resin/dentin bond strength. AB - PURPOSE: This study aimed to compare the effect of three curing modes of a high powered curing-light source on the shear bond strength and marginal gap of light- and dual-cured adhesive resin cements to dentin. MATERIALS AND METHODS: Twelve freshly extracted intact human mandibular molars were selected for this study and stored in the saline solution. Three of the axial surfaces of the teeth were prepared to obtain flat dentinal surfaces. Thirty-six ceramic disks (4-mm diameter, 2- mm thick) were constructed from a pressable glass-ceramic (Vision). The discs were etched with hydrofluoric acid and primed, and then divided into two equal groups, groups I and II (n = 18 each). Two adhesive systems were used following manufacturer's instructions. The discs of group I were bonded to the conditioned dentin surface using adhesive resin (Rely X Veneer), and group II discs were bonded to dentin using Rely X ARC. For each group, the resin was cured using three modes (fast, ramp, pulse). Interfacial gap at the dentin/resin interface was measured at eight predetermined sites for each specimen using a stereomicroscope, and shear bond strength of the bonded specimens was carried out using a universal testing machine. RESULTS: Ramp-cured specimens recorded significantly higher mean shear bond strengths for both dual- and light polymerized resins than those with fast and pulse modes. Moreover, fewer interfacial gaps were found at the resin/dentin interface in association with ramp cure modes of both resins. Most failures were adhesive failures at the dentin-resin luting agent (RLA) interface in specimens polymerized using high powered LED fast or pulse modes, while a cohesive failure pattern within the resin was associated with the ramp-curing mode. CONCLUSION: Within the limitations of this study, the shear bond strength of an RLA to dentin was found to be enhanced with light- or dual-polymerized adhesive resin using an LED light in ramp mode, whereas shear bond strength was significantly lower when polymerized using LED in fast or pulse modes. PMID- 19682223 TI - Left brain/right brain in sexual medicine. PMID- 19682225 TI - Incision and grafting for severe Peyronie's disease (CME). AB - BACKGROUND: Peyronie's Disease is a wound healing disorder involving growth of fibrous plaques in the tunica albuginea, and often results in abnormal penile curvature and subsequent development of erectile dysfunction. A 59-year-old man with a history of untreated penile trauma who presented with a 6-year history of worsening penile curvature that interfered with sexual functioning and resulted in associated erectile dysfunction. METHODS: The patient's Peyronie's Disease was evaluated in clinic with a focused physical exam and a penile vasculature study using a color duplex Doppler ultrasound. Since the patient did not have proper insurance coverage for an inflatable penile prosthesis, but did have normal arterial flow with only boderline veno-occlusive disease, he instead underwent an incision and grafting procedure. After degloving the penis, a lateral approach parallel to the corpus spongiosum was used to enter Buck's fascia and isolate the neurovascular bundles. The fibrous plaque was incised with a residual tunical defect of 4 x 5 cm, and covered with a pericardial graft. RESULTS: Preoperatively, artificially induced erection during the patient's Doppler study demonstrated dorsal curvature greater than 80 degrees. Intraoperatively, after completing the incision and grafting procedure, artificial erection demonstrated minimal (less than 15 degrees) residual curvature. At his 1-week postoperative visit, he reported spontaneous erections and minimal pain. One month postoperatively, his incisions were well healed and an artificially induced erection continued to demonstrate minimal curvature. DISCUSSION: Management of severe Peyronie's Disease with significant penile curvature is a challenging clinical problem. In extreme cases, placement of an inflatable penile prosthesis with ancillary techniques usually gives the best clinical outcome. Although more difficult to execute in severe cases, incision and grafting represents an acceptable alternative in the appropriately selected patient. PMID- 19682228 TI - Further experience with an operation for the cure of certain types of impotence. O.S. Lowsley, and A. Rueda. PMID- 19682231 TI - Retrograde effects of triazolam and zolpidem on sleep-dependent motor learning in humans. AB - Drugs that act as allosteric activators at the benzodiazepine site of the gamma aminobutyric acid (GABA(A)) receptor complex are used commonly to treat insomnia but relatively little is known of how such use affects learning and memory. Although anterograde effects on memory acquisition have been shown, possible retrograde effects on consolidation are more relevant when such agents are administered at bedtime. We tested the effects of two GABA(A) allosteric activators on sleep-dependent motor skill memory consolidation in 12 healthy male subjects. Subjects slept in a sleep laboratory for four consecutive nights (one accommodation night followed by three experimental nights). Placebo, triazolam 0.375 mg, and zolpidem 10 mg were given to each subject in counterbalanced order on the experimental nights. Polysomnographic (PSG) sleep measurement and sleep dependent motor learning were assessed at each condition. Triazolam was associated with longer total sleep time and increased Stage 2 sleep. Both zolpidem and triazolam were associated with increased latency to rapid eye movement (REM) sleep. Overnight motor learning correlated with total sleep time in the placebo condition but not in the triazolam or zolpidem conditions. A statistically significant impairment in motor performance occurred overnight in the triazolam condition only. Triazolam, given in sufficient doses to prolong sleep in healthy people, affected overnight motor learning adversely. Zolpidem, in a dose sufficient to prolong REM onset latency but without other effects on PSG-measured sleep, degraded the relationship between total sleep time and overnight motor learning. These data indicate that non-selective or alpha1 preferring benzodiazepine site allosteric activators can interfere with sleep dependent memory consolidation. PMID- 19682232 TI - Circulating inflammatory and hemostatic biomarkers are associated with risk of myocardial infarction and coronary death, but not angina pectoris, in older men. AB - AIMS: The extent to which hemostatic and inflammatory biomarkers are related to angina pectoris as compared with myocardial infarction (MI) remains uncertain. We examined the relationship between a wide range of inflammatory and hemostatic biomarkers, including markers of activated coagulation, fibrinolysis and endothelial dysfunction and viscosity, with incident myocardial infarction (MI) or coronary heart disease (CHD) death and incident angina pectoris uncomplicated by MI or CHD death in older men. METHODS: A prospective study of 3217 men aged 60 79 years with no baseline CHD (angina or MI) and who were not on warfarin, followed up for 7 years during which there were 198 MI/CHD death cases and 220 incident uncomplicated angina cases. RESULTS: Inflammatory biomarkers [C-reactive protein (CRP), interleukin-6, fibrinogen], plasma viscosity and hemostatic biomarkers [von Willebrand factor (VWF) and fibrin D-dimer] were associated with a significant increased risk of MI/CHD death but not with uncomplicated angina even after adjustment for age and conventional risk factors. Adjustment for CRP attenuated the relationships between VWF, fibrin D-dimer and plasma viscosity with MI/CHD death. Comparisons of differing associations with risk of MI/CHD deaths and uncomplicated angina were significant for the inflammatory markers (P < 0.05) and marginally significant for fibrin D-dimer (P = 0.05). In contrast, established risk factors including blood pressure and high-density lipoprotein (HDL)-cholesterol were associated with both MI/CHD death and uncomplicated angina. CONCLUSION: Circulating biomarkers of inflammation and hemostasis are associated with incident MI/CHD death but not incident angina uncomplicated by MI or CHD death in older men. PMID- 19682233 TI - Marked reduction of early stent thrombosis with pre-hospital initiation of high dose Tirofiban in ST-segment elevation myocardial infarction. AB - BACKGROUND: No randomized comparisons are yet available evaluating the effect of pre-hospital high dose tirofiban on the incidence of early stent thrombosis after primary percutaneous coronary intervention (PCI). OBJECTIVES: The aim of this analysis was to evaluate whether routine pre-hospital administration of high-dose tirofiban in ST-segment elevation myocardial infarction (STEMI) decreases the incidence of early stent thrombosis after primary PCI. PATIENTS/METHODS: The Ongoing Tirofiban in Myocardial Evaluation (On-TIME) 2 trial was a prospective multicenter study of consecutive STEMI patients referred for primary PCI in which patients were randomized to pre-hospital no high-dose tirofiban/placebo. We examined the incidence of Academic Research Consortium definite and probable early stent thrombosis and determined predictors and outcome of early stent thrombosis. RESULTS: Primary PCI was performed in 1203 out of 1398 patients (86.1%). In 1073 patients (89.2%) a coronary stent was placed. Early stent thrombosis occurred in 39 patients (3.6%). Pre-hospital initiation of high-dose tirofiban significantly reduced early stent thrombosis (2.1% vs. 5.2%, P = 0.006) and was associated with a lower incidence of urgent repeat PCI (1.9% vs. 5.2%, P = 0.005). Early stent thrombosis, as well as pre-hospital initiation of high-dose tirofiban, was independently associated with 30-day mortality. CONCLUSIONS: Pre hospital initiation of high-dose tirofiban reduces the 30-day incidence of stent thrombosis in STEMI patients treated with primary PCI and stenting. Early stent thrombosis and pre-hospital initiation of high-dose tirofiban were independent predictors of 30-day mortality. PMID- 19682234 TI - Clinical probability score and D-dimer estimation lack utility in the diagnosis of childhood pulmonary embolism. AB - BACKGROUND: Childhood pulmonary embolism (PE) causes significant mortality and evidence suggests that it is under-diagnosed. Clinical probability scores and D dimer estimation to assess pre-test probability have not been studied in children with suspected PE. PATIENTS/METHODS: This retrospective cohort study evaluated Wells simplified probability score for PE in 50 children with PE and 25 PE negative control patients, and D-dimer values in 27 PE positive and 12 PE negative children. RESULTS: PE positive and PE negative groups had similar rates of risk factors for venous thromboembolism (VTE). Wells simplified probability score showed a small difference between PE positive and PE negative children (median score: PE positive, 4.5; PE negative, 4; P = 0.009), children with PE are more likely to obtain a 'PE likely' score (score > 4), P = 0.012. The difference was of slightly greater significance when the Wells score was adjusted to account for pediatric normal ranges for heart rate, P = 0.007, and signs/symptoms of upper limb DVT, P = 0.006. Children with PE were as likely as PE negative patients to have a D-dimer value within the normal range (PE positive, 15%; PE negative, 25%; P = 0.654). A combination of a 'PE unlikely' score and normal D dimer value occurred in 1/12 (8%) of PE negative children. CONCLUSIONS: The Wells clinical probability score and D-dimer estimation may lack utility in the determination of pre-test probability of PE in children. Validation of a pediatric clinical probability score, incorporating D-dimer estimation, by prospective study, would be difficult as a result of the rarity of childhood PE. PMID- 19682235 TI - Splenic marginal zone antigen-presenting cells are critical for the primary allo immune response to therapeutic factor VIII in hemophilia A. AB - BACKGROUND: Alloimmune responses to intravenously administered protein therapeutics are the most common cause of failure of replacement therapy in patients with defective levels of endogenous proteins. Such a situation is encountered in some patients with hemophilia A, who develop inhibitory anti factor (F)VIII alloantibodies after administration of FVIII to treat hemorrhages. OBJECTIVES: The nature of the secondary lymphoid organs involved in the initiation of immune responses to human therapeutic has not been studied. We therefore investigated this in the case of FVIII, a self-derived exogenous protein therapeutic. METHODS: The distribution of intravenously administered FVIII was followed after FVIII-deficient mice were injected with radiolabeled FVIII and using immunohistochemistry. The role of the spleen and antigen presenting cells (APC) in the onset of the anti-FVIII immune response was analyzed upon splenectomy or treatment of the mice with APC-depleting compounds. RESULTS: FVIII preferentially accumulated in the spleen at the level of metallophilic macrophages in the marginal zone (MZ). Surgical removal of the spleen or selective in vivo depletion of macrophages and CD11c-positive CD8 alpha negative dendritic cells resulted in a drastic reduction in anti-FVIII immune responses. CONCLUSIONS: Using FVIII-deficient mice as a model for patients with hemophilia A, and human pro-coagulant FVIII as a model for immunogenic self derived protein therapeutics, our results highlight the importance of the spleen and MZ APCs in the initiation of immune responses to protein therapeutics. Identification of the receptors implicated in retention of protein therapeutics in the MZ may pave the way towards novel strategies aimed at reducing their immunogenicity. PMID- 19682236 TI - von Willebrand factor cleaved from endothelial cells by ADAMTS13 remains ultralarge in size. PMID- 19682237 TI - Total knee arthroplasty in hemophilic arthropathy: efficiency of clotting factor usage in multijoint procedures. PMID- 19682238 TI - IgG subclass distribution of anti-ADAMTS13 antibodies in patients with acquired thrombotic thrombocytopenic purpura. AB - BACKGROUND: ADAMTS13-neutralizing IgG autoantibodies are the major cause of acquired thrombotic thrombocytopenic purpura (TTP). OBJECTIVE: To analyze the IgG subclass distribution of anti-ADAMTS13 antibodies and a potential relationship between subclass distribution and disease prognosis. METHODOLOGY: An enzyme linked immunosorbent assay-based method was used to quantify the relative amounts of IgG subclasses of anti-ADAMTS13 antibodies in acquired TTP plasma. RESULTS: IgG(4) (52/58, 90%) was the most prevalent IgG subclass in patients with acquired TTP, followed by IgG(1) (52%), IgG(2) (50%), and IgG(3) (33%). IgG(4) was found either alone (17/52) or with other IgG subclasses (35/52). IgG(4) was not detected in 10% of the patients. There was an inverse correlation between the frequency and abundance of IgG(4) and IgG(1) antibodies (P < 0.01). Patients with high IgG(4) levels and undetectable IgG(1) are more prone to relapse than patients with low IgG(4) levels and detectable IgG(1). CONCLUSIONS: All IgG subclasses of anti-ADAMTS13 antibodies were detected in patients with acquired TTP, with IgG(4), followed by IgG(1), antibodies dominating the anti-ADAMTS13 immune response. Levels of IgG(4) could be useful for the identification of patients at risk of disease recurrence. PMID- 19682239 TI - Replication of findings on the association of genetic variation in 24 hemostasis genes and risk of incident venous thrombosis. PMID- 19682240 TI - Platelet glycoprotein VI as a mediator of metastasis. PMID- 19682241 TI - Proteomic analysis of integrin alphaIIbbeta3 outside-in signaling reveals Src kinase-independent phosphorylation of Dok-1 and Dok-3 leading to SHIP-1 interactions. AB - BACKGROUND AND OBJECTIVES: Outside-in integrin alphaIIbbeta3 signaling involves a series of tyrosine kinase reactions that culminate in platelet spreading on fibrinogen. The aim of this study was to identify novel tyrosine phosphorylated signaling proteins downstream of alphaIIbbeta3, and explore their role in platelet signaling. METHODS AND RESULTS: Utilizing proteomics to search for novel platelet proteins that contribute to outside-in signaling by the integrin alphaIIbbeta3, we identified 27 proteins, 17 of which were not previously shown to be part of a tyrosine phosphorylation-based signaling complex downstream of alphaIIbbeta3. The proteins identified include the novel immunoreceptors G6f and G6b-B, and two members of the Dok family of adapters, Dok-1 and Dok-3, which underwent increased tyrosine phosphorylation following platelet spreading on fibrinogen. Dok-3 was also inducibly phosphorylated in response to the GPVI specific agonist collagen-related peptide (CRP) and the PAR-1 and -4 agonist thrombin, independently of the integrin alphaIIbbeta3. Tyrosine phosphorylation of Dok-1 and Dok-3 was primarily Src kinase-independent downstream of the integrin, whereas it was Src kinase-dependent downstream of GPVI. Moreover, both proteins inducibly interacted with Grb-2 and SHIP-1 in fibrinogen-spread platelets. CONCLUSIONS: This study provides new insights into the molecular mechanism regulating alphaIIbbeta3-mediated platelet spreading on fibrinogen. The novel platelet adapter Dok-3 and the structurally related Dok-1 are tyrosine phosphorylated in an Src kinase-independent manner downstream of alphaIIbbeta3 in human platelets, leading to an interaction with Grb2 and SHIP-1. PMID- 19682242 TI - Sleep state distribution of obstructive events in children: is obstructive sleep apnoea really a rapid eye movement sleep-related condition? AB - Obstructive sleep apnoea (OSA) in children is commonly considered to occur predominantly in rapid eye movement (REM) sleep, but clinical experience suggests that this is not universally the case. We hypothesized that there would be a subgroup of children with OSA who have non-REM (NREM) predominance of obstructive events and that these children share certain clinical characteristics. Thus, we aimed to compare the obstructive apnoea-hypopnoea index (OAHI) in REM versus NREM sleep and to assess factors influencing the distribution of events by sleep state. Polysomnography (PSG) recordings of 102 children aged 0-18 years with moderate to severe OSA (OAHI >or=5 h(-1)) were reviewed. OAHI was calculated separately for REM and NREM sleep. A REM predominance index (RPI) was determined using log transformation [RPI = log (REM OAHI + 0.5) - log (NREM OAHI + 0.5)] and compared with possible influencing factors using multiple linear regression. Analysis showed that obstructive events were more common in REM sleep (median REM OAHI 21.4 h(-1), median NREM OAHI 8.3 h(-1), P < 0.001). Mean RPI was significantly greater than zero (P = 0.003). However, a substantial minority of children (30.4%) had a higher NREM than REM OAHI. The factors that were related significantly to NREM predominance were older age (P = 0.02), higher arousal index (P < 0.001) and higher SpO(2) nadir (P < 0.001). Our findings demonstrate that while OSA is a REM sleep-related problem in the majority of children, there is a significant subset of children with NREM predominance of obstructive events. This finding highlights the importance of considering sleep state distribution of events in studies of the pathophysiology and outcomes of OSA in childhood. PMID- 19682243 TI - Factor structure of the Athens Insomnia Scale and its associations with demographic characteristics and depression in adolescents. AB - The eight-item Athens Insomnia Scale (AIS-8) is an instrument that has been used frequently to assess insomnia problems. Previous research on adults has found that the AIS-8 functioned as a sole component. This study aimed to examine the prevalence rates of insomnia problems on the AIS-8, the factor structure of the AIS-8 in adolescents and its associations with demographic characteristics and depression in adolescents. A total of 8319 adolescent students (4334 girls and 3985 boys, mean age = 14.7 years, standard deviation = 1.7 years) in southern Taiwan were recruited into this study and completed the AIS-8. We performed an exploratory factor analysis to examine the factor structure of the AIS-8, and used the parallel analysis for making decisions regarding factor retention. We also used multiple regression analysis models to determine the associations between insomnia and demographic characteristics and depression. The results found that a high proportion of adolescents had insomnia problems as measured by the AIS-8. The AIS-8 was composed of two different factors when used among a large adolescent population, including insomnia symptoms (factor 1) and subjective sleep and daytime distress (factor 2). While being male, being younger, and having depression were associated positively with the severity of insomnia symptoms (factor 1), being older, living in urban areas, and having depression were associated positively with the severity of subjective sleep and daytime distress (factor 2). Clinicians and researchers should consider the different meanings of the two factors of the AIS-8 when using this tool to assess insomnia problems in adolescents. PMID- 19682244 TI - Complexity and context of MHC-correlated mating preferences in wild populations. AB - There is now substantial and growing evidence for a role of the major histocompatibility complex (MHC) in shaping individual mate preferences. In view of both its codominant expression and its function in immune response, it is often expected that females aim to avoid inbreeding or maximize offspring MHC heterozygosity by selecting as mates those males which share fewest or no MHC alleles with themselves. However, it is becoming increasingly clear that this view is over-simplistic: not only is MHC dissimilarity just one of several (perhaps many) criteria important in mate choice decision-making, extremely MHC dissimilar males may be avoided, and furthermore, specific alleles or combinations might be preferred if they bestow particular advantages. These points are raised in two papers in this Molecular Ecology issue, in which patterns of reproductive success in tiger salamanders (Ambystoma tigrinum) and three-spined sticklebacks (Gasterosteus aculeatus) are each inconsistent with a generalized preference for MHC dissimilarity. Together, these studies demonstrate several adaptive reasons for decision rules that do not necessarily result in maximizing mate dissimilarity. PMID- 19682245 TI - An alternative to the glyoxylate shunt. AB - A cycle remains a cycle only as long as the spokes of the wheel are not stolen. To keep the citric acid cycle going requires anaplerotic reactions such as the glyoxylate shunt to restore the cycle intermediates that are withdrawn for the biosynthesis of cell constituents, e.g. amino acids and haemin precursors. The article by Erb et al. in this issue of Molecular Microbiology documents an alternative path that replenishes four-carbon intermediates during growth on acetate in the absence of the glyoxylate shunt. The reaction sequence forms malate and succinyl-CoA from three acetyl-CoA, one CO(2) and one HCO(3) in a linear pathway. This new pathway was discovered in phototrophic anoxygenic bacteria and in few aerobic bacteria, but it is probably widespread among many metabolic groups of bacteria. PMID- 19682246 TI - Co-ordinated regulation of two divergent promoters through higher-order complex formation by the LysR-type regulator ThnR. AB - The genes required for tetralin biodegradation by Sphingomonas macrogolitabida strain TFA are clustered in two divergent and closely linked operons. ThnR, a LysR-type regulator, activates transcription from each operon in response to tetralin. The regulatory thnR gene is co-transcribed with the catabolic genes thnC, thnA3 and thnA4, resulting in positive autoregulation. ThnR binds with different affinity to two primary binding sites, designated B and C, in the intervening region between the two operons and makes additional contact with secondary sites that extend towards the promoters. In addition, ThnR may interact with itself when bound to each site via the formation of a DNA loop, as evidenced by the distortion of the DNA between the primary binding sites and the elimination of the higher-order complexes following the introduction of a half turn of the DNA helix between the primary binding sites. Transcription from each promoter is not fully independent since mutations in each binding site affected transcription from both promoters. Based on these results, we propose a model of transcription activation that involves the formation of a complex structure by interactions between ThnR molecules bound to distant binding sites and favours transcription from one promoter to the detriment of the other. PMID- 19682247 TI - A mechanism of transposon-mediated directed mutation. AB - Directed mutation is a proposed process that allows mutations to occur at higher frequencies when they are beneficial. Until now, the existence of such a process has been controversial. Here we describe a novel mechanism of directed mutation mediated by the transposon, IS5 in Escherichia coli. crp deletion mutants mutate specifically to glycerol utilization (Glp(+)) at rates that are enhanced by glycerol or the loss of the glycerol repressor (GlpR), depressed by glucose or glpR overexpression, and RecA-independent. Of the four tandem GlpR binding sites (O1-O4) upstream of the glpFK operon, O4 specifically controls glpFK expression while O1 primarily controls mutation rate in a process mediated by IS5 hopping to a specific site on the E. coli chromosome upstream of the glpFK promoter. IS5 insertion into other gene activation sites is unaffected by the presence of glycerol or the loss of GlpR. The results establish an example of transposon mediated directed mutation, identify the protein responsible and define the mechanism involved. PMID- 19682248 TI - The transcriptionally active regions in the genome of Bacillus subtilis. AB - The majority of all genes have so far been identified and annotated systematically through in silico gene finding. Here we report the finding of 3662 strand-specific transcriptionally active regions (TARs) in the genome of Bacillus subtilis by the use of tiling arrays. We have measured the genome-wide expression during mid-exponential growth on rich (LB) and minimal (M9) medium. The identified TARs account for 77.3% of the genes as they are currently annotated and additionally we find 84 putative non-coding RNAs (ncRNAs) and 127 antisense transcripts. One ncRNA, ncr22, is predicted to act as a translational control on cstA and an antisense transcript was observed opposite the housekeeping sigma factor sigA. Through this work we have discovered a long conserved 3' untranslated region (UTR) in a group of membrane-associated genes that is predicted to fold into a large and highly stable secondary structure. One of the genes having this tail is efeN, which encodes a target of the twin-arginine translocase (Tat) protein translocation system. PMID- 19682249 TI - Two-enzyme systems for glycolipid and polyglycerolphosphate lipoteichoic acid synthesis in Listeria monocytogenes. AB - Lipoteichoic acid (LTA) is an important cell wall polymer in gram-positive bacteria and often consists a polyglycerolphosphate backbone chain that is linked to the membrane by a glycolipid. In Listeria monocytogenes this glycolipid is Gal Glc-DAG or Gal-Ptd-6Glc-DAG. Using a bioinformatics approach, we have identified L. monocytogenes genes predicted to be involved in glycolipid (lmo2555 and lmo2554) and LTA backbone (lmo0644 and lmo0927) synthesis. LTA and glycolipid analysis of wild-type and mutant strains confirmed the function of Lmo2555 and Lmo2554 as glycosyltransferases required for the formation of Glc-DAG and Gal-Glc DAG. Deletion of a third gene, lmo2553, located in the same operon resulted in the production of LTA with an altered structure. lmo0927 and lmo0644 encode proteins with high similarity to the staphylococcal LTA synthase LtaS, which is responsible for polyglycerolphosphate backbone synthesis. We show that both proteins are involved in LTA synthesis. Our data support a model whereby Lmo0644 acts as an LTA primase LtaP and transfers the initial glycerolphosphate onto the glycolipid anchor, and Lmo0927 functions as LTA synthase LtaS, which extends the glycerolphosphate backbone chain. Inactivation of LtaS leads to severe growth and cell division defects, underscoring the pivotal role of LTA in this gram-positive pathogen. PMID- 19682250 TI - Vital role for the Plasmodium actin capping protein (CP) beta-subunit in motility of malaria sporozoites. AB - Summary Successful malaria transmission from the mosquito vector to the mammalian host depends crucially on active sporozoite motility. Sporozoite locomotion and host cell invasion are driven by the parasite's own actin/myosin motor. A unique feature of this motor machinery is the presence of very short subpellicular actin filaments. Therefore, F-actin stabilizing proteins likely play a central role in parasite locomotion. Here, we investigated the role of the Plasmodium berghei actin capping protein (PbCP), an orthologue of the heterodimeric regulator of filament barbed end growth, by reverse genetics. Parasites containing a deletion of the CP beta-subunit developed normally during the pathogenic erythrocytic cycle. However, due to reduced ookinete motility, mutant parasites form fewer oocysts and sporozoites in the Anopheles vector. These sporozoites display a vital deficiency in forward gliding motility and fail to colonize the mosquito salivary glands, resulting in complete attenuation of life cycle progression. Together, our results show that the CP beta-subunit exerts an essential role in the insect vector before malaria transmission to the mammalian host. The vital role is restricted to fast locomotion, as displayed by Plasmodium sporozoites. PMID- 19682251 TI - Enhancer of decapping proteins 1 and 2 are important for translation during heat stress in Saccharomyces cerevisiae. AB - In mammalian and Drosophila cells, heat stress strongly reduces general protein translation while activating cap-independent translation mechanisms to promote the expression of stress-response proteins. In contrast, in Saccharomyces cerevisiae general translation is only mildly and transiently reduced by heat stress and cap-independent translation mechanisms have not been correlated with the heat stress response. Recently we have identified direct target genes of the heat shock transcription factor (HSF), including genes encoding proteins thought to be important for general translation. One gene activated by HSF during heat stress encodes the enhancer of decapping protein, Edc2, previously shown to enhance mRNA decapping under conditions when the decapping machinery is limited. In this report we show that strains lacking Edc2, as well as the paralogous protein Edc1, are compromised for growth under persistent heat stress. This growth deficiency can be rescued by expression of a mutant Edc1 protein deficient in mRNA decapping indicative of a decapping independent function during heat stress. Yeast strains lacking Edc1 and Edc2 are also sensitive to the pharmacological inhibitor of translation paromomycin and exposure to heat stress and paromomycin functions synergistically to reduce yeast viability, suggesting that in the absence of Edc1 and Edc2 translation is compromised under heat stress conditions. Strains lacking Edc1 and Edc2 have significantly reduced rates of protein translation during growth under heat stress conditions, but not under normal growth conditions. We propose that Edc1 and the stress responsive isoform Edc2 play important roles in protein translation during stress. PMID- 19682252 TI - SirA enforces diploidy by inhibiting the replication initiator DnaA during spore formation in Bacillus subtilis. AB - How cells maintain their ploidy is relevant to cellular development and disease. Here, we investigate the mechanism by which the bacterium Bacillus subtilis enforces diploidy as it differentiates into a dormant spore. We demonstrate that a sporulation-induced protein SirA (originally annotated YneE) blocks new rounds of replication by targeting the highly conserved replication initiation factor DnaA. We show that SirA interacts with DnaA and displaces it from the replication origin. As a result, expression of SirA during growth rapidly blocks replication and causes cell death in a DnaA-dependent manner. Finally, cells lacking SirA over-replicate during sporulation. These results support a model in which induction of SirA enforces diploidy by inhibiting replication initiation as B. subtilis cells develop into spores. PMID- 19682253 TI - Positive and negative feedback regulatory loops of thiol-oxidative stress response mediated by an unstable isoform of sigmaR in actinomycetes. AB - Alternate sigma factors provide an effective way of diversifying bacterial gene expression in response to environmental changes. In Streptomyces coelicolor where more than 65 sigma factors are predicted, sigma(R) is the major regulator for response to thiol-oxidative stresses. sigma(R) becomes available when its bound anti-sigma factor RsrA is oxidized at sensitive cysteine thiols to form disulphide bonds. sigma(R) regulon includes genes for itself and multiple thiol reducing systems, which constitute positive and negative feedback loops respectively. We found that the positive amplification loop involves an isoform of sigma(R) (sigma(R')) with an N-terminal extension of 55 amino acids, produced from an upstream start codon. A major difference between constitutive sigma(R) and inducible sigma(R') is that the latter is markedly unstable (t(1/2) approximately 10 min) compared with the former (> 70 min). The rapid turnover of sigma(R') is partly due to induced ClpP1/P2 proteases from the sigma(R) regulon. This represents a novel way of elaborating positive and negative feedback loops in a control circuit. Similar phenomenon may occur in other actinomycetes that harbour multiple start codons in the sigR homologous gene. We observed that sigH gene, the sigR orthologue in Mycobacterium smegmatis, produces an unstable larger isoform of sigma(H) upon induction by thiol-oxidative stress. PMID- 19682255 TI - Genetic analysis of xenocoumacin antibiotic production in the mutualistic bacterium Xenorhabdus nematophila. AB - Xenocoumacin 1 (Xcn1) and xenocoumacin 2 (Xcn2) are the major antimicrobial compounds produced by Xenorhabdus nematophila. To study the role of Xcn1 and Xcn2 in the life cycle of X. nematophila the 14 gene cluster (xcnA-N) required for their synthesis was identified. Overlap RT-PCR analysis identified six major xcn transcripts. Individual inactivation of the non-ribosomal peptide synthetase genes, xcnA and xcnK, and polyketide synthetase genes, xcnF, xcnH and xcnL, eliminated Xcn1 production. Xcn1 levels and expression of xcnA-L were increased in an ompR strain while Xcn2 levels and xcnMN expression were reduced. Xcn1 production was also increased in a strain lacking acetyl-phosphate that can donate phosphate groups to OmpR. Together these findings suggest that OmpR phosphate negatively regulates xcnA-L gene expression while positively regulating xcnMN expression. HPLC-MS analysis revealed that Xcn1 was produced first and was subsequently converted to Xcn2. Inactivation of xcnM and xcnN eliminated conversion of Xcn1 to Xcn2 resulting in elevated Xcn1 production. The viability of the xcnM strain was reduced 20-fold relative to the wild-type strain supporting the idea that conversion of Xcn1 to Xcn2 provides a mechanism to avoid self-toxicity. Interestingly, inactivation of ompR enhanced cell viability during prolonged culturing. PMID- 19682254 TI - ESX-1 secreted virulence factors are recognized by multiple cytosolic AAA ATPases in pathogenic mycobacteria. AB - The ESX-1 secretion system of Mycobacterium tuberculosis delivers bacterial virulence factors to host cells during infection. The most abundant factor, the ESAT-6/CFP-10 dimer, is targeted for secretion via a C-terminal signal sequence on CFP-10 that is recognized by the cytosolic ATPase, Rv3871. However, the selection determinants for other ESX-1 substrates appear to be more complex. Some substrates, such as ESAT-6, are secreted despite lacking signal sequences. Furthermore, all substrates require targeting of the other ESX-1 secreted proteins, a distinguishing feature of this system. How ESX-1 substrates are selected and the basis for co-dependent secretion is unknown. Here we show that the EspC substrate interacts with Rv3868, a cytosolic AAA ATPase, through its C terminus. Swapping the C-termini of EspC and CFP-10 revealed that these signals are functionally distinct, suggesting that the proteins are targeted via interactions with different ATPases. Surprisingly, biochemical purification experiments demonstrate that these substrates and ATPases form multi-protein complexes inside the cell and identified a new secreted substrate. By interfering with this protein interaction network, we have partially uncoupled co-dependent substrate secretion. Our results suggest that proper functioning of the ESX-1 pathway requires the interaction of multiple ESX-1 substrates and components prior to their secretion. Ultimately, understanding the details of ESX-1 targeting may allow for engineering of better vaccines to prevent tuberculosis. PMID- 19682256 TI - Physical, functional and conditional interactions between ArcAB and phage shock proteins upon secretin-induced stress in Escherichia coli. AB - The phage shock protein (Psp) system found in enterobacteria is induced in response to impaired inner membrane integrity (where the Psp response is thought to help maintain the proton motive force of the cell) and is implicated in the virulence of pathogens such as Yersinia and Salmonella. We provided evidence that the two-component ArcAB system was involved in induction of the Psp response in Escherichia coli and now report that role of ArcAB is conditional. ArcAB, predominantly through the action of ArcA regulated genes, but also via a direct ArcB-Psp interaction, is required to propagate the protein IV (pIV)-dependent psp inducing signal(s) during microaerobiosis, but not during aerobiosis or anaerobiosis. We show that ArcB directly interacts with the PspB, possibly by means of the PspB leucine zipper motif, thereby allowing cross-communication between the two systems. In addition we demonstrate that the pIV-dependent induction of psp expression in anaerobiosis is independent of PspBC, establishing that PspA and PspF can function as a minimal Psp system responsive to inner membrane stress. PMID- 19682257 TI - Decreased outer membrane permeability protects mycobacteria from killing by ubiquitin-derived peptides. AB - Ubiquitin-derived peptides are bactericidal in vitro and contribute to the mycobactericidal activity of the lysosome. To further define interactions of ubiquitin-derived peptides with mycobacteria, we screened for mutants with increased resistance to the bactericidal activity of the synthetic ubiquitin derived peptide Ub2. The four Ub2-resistant Mycobacterium smegmatis mutants were also resistant to the bactericidal action of other antimicrobial peptides and macrophages. Two mutants were in the mspA gene encoding the main M. smegmatis porin. Using a translocation-deficient MspA point mutant, we showed that susceptibility of M. smegmatis to Ub2 was independent of MspA channel activity. Instead, the M. smegmatis Ub2-resistant mutants shared a common phenotype of decreased cell wall permeability compared with wild-type bacteria. Expression of mspA rendered Mycobacterium tuberculosis CDC1551 more susceptible both to ubiquitin-derived peptides in vitro and to lysosomal killing in macrophages. Finally, biochemical assays designed to assess membrane integrity indicated that Ub2 treatment impairs membrane function of M. smegmatis and M. tuberculosis cells. The M. smegmatis Ub2-resistant mutants were more resistant than wild-type M. smegmatis to this damage. We conclude that Ub2 targets mycobacterial membranes and that reduced membrane permeability provides mycobacteria intrinsic resistance against antimicrobial compounds including bactericidal ubiquitin-derived peptides. PMID- 19682258 TI - A LuxRI-family regulatory system controls excision and transfer of the Mesorhizobium loti strain R7A symbiosis island by activating expression of two conserved hypothetical genes. AB - The symbiosis island ICEMlSym(R7A) of Mesorhizobium loti R7A is an integrative and conjugative element (ICE) that carries genes required for a nitrogen-fixing symbiosis with Lotus species. ICEMlSym(R7A) encodes homologues (TraR, TraI1 and TraI2) of proteins that regulate plasmid transfer by quorum sensing in rhizobia and agrobacteria. Introduction of traR cloned on a plasmid induced excision of ICEMlSym(R7A) in all cells, a 1000-fold increase in the production of 3-oxo-C6 homoserine lactone (3-oxo-C6-HSL) and a 40-fold increase in conjugative transfer. These effects were dependent on traI1 but not traI2. Induction of expression from the traI1 and traI2 promoters required the presence of plasmid-borne traR and either traI1 or 100 pM 3-oxo-C6-HSL, suggesting that traR expression or TraR activity is repressed in wild-type cells by a mechanism that can be overcome by additional copies of traR. The traI2 gene formed an operon with hypothetical genes msi172 and msi171 that were essential for ICEMlSym(R7A) excision and transfer. Our data suggest that derepressed TraR in conjunction with TraI1 synthesized 3-oxo-C6-HSL regulates excision and transfer of ICEMlSym(R7A) through expression of msi172 and msi171. Homologues of msi172 and msi171 were present on putative ICEs in several alpha-proteobacteria, indicating a conserved role in ICE excision and transfer. PMID- 19682259 TI - Cation/proton antiporter complements of bacteria: why so large and diverse? AB - Most bacterial genomes have five to nine distinct genes predicted to encode transporters that exchange cytoplasmic Na(+) and/or K(+) for H(+) from outside the cell, i.e. monovalent cation/proton antiporters. By contrast, pathogens that live primarily inside host cells usually possess zero to one such antiporter while other stress-exposed bacteria exhibit even higher numbers. The monovalent cation/proton antiporters encoded by these diverse genes fall into at least eight different transporter protein families based on sequence similarity. They enable bacteria to meet challenges of high or fluctuating pH, salt, temperature or osmolarity, but we lack explanations for why so many antiporters are needed and for the value added by specific antiporter types in specific settings. In this issue of Molecular Microbiology, analyses of the pH dependence of cytoplasmic [Na(+)], [K(+)], pH and transmembrane electrical potential in the 'poly extremophile'Natranaerobius thermophilus are the context for assessment of the catalytic properties of 12 predicted monovalent cation/proton antiporters in the genome of this thermophilic haloalkaliphile. The results provide a profile of adaptations of the poly extremophilic anaerobe, including a proposed role of cytoplasmic buffering capacity. They also provide new perspectives on two large monovalent cation/proton antiporter families, the NhaC and the cation/proton antiporter-3 antiporter families. PMID- 19682260 TI - Another way to divide: the case of anammox bacteria. AB - Anammox bacteria exhibit remarkable and characteristic metabolic features that enable them to oxidize ammonium under anoxic conditions. Their ultrastructure is complex, with three compartments, comprising paryphoplasm, the riboplasm and the anammoxosome. The latter is surrounded by ladderane lipids, creating a lipid surrounded organelle within the riboplasm. In this edition of Molecular Microbiology, van Niftrik and co-workers present detailed ultrastructural investigations that, together with the analysis of the product of a predicted gene and its localization in ultrathin sections, provide the first hints at the way in which these cells divide. PMID- 19682261 TI - Attachment of Streptomyces coelicolor is mediated by amyloidal fimbriae that are anchored to the cell surface via cellulose. AB - The chaplin proteins ChpA-H enable the filamentous bacterium Streptomyces coelicolor to form reproductive aerial structures by assembling into surface active amyloid-like fibrils. We here demonstrate that chaplins also mediate attachment of S. coelicolor to surfaces. Attachment coincides with the formation of fimbriae, which are connected to the cell surface via spike-shaped protrusions. Mass spectrometry, electron microscopy and Congo red treatment showed that these fimbriae are composed of bundled amyloid fibrils of chaplins. Attachment and fimbriae formation were abolished in a strain in which the chaplin genes chpA-H were inactivated. Instead, very thin fibrils emerged from the spike shaped protrusions in this mutant. These fibrils were susceptible to cellulase treatment. This enzymatic treatment also released wild-type fimbriae from the cell surface, thereby abolishing attachment. The reduced attachment of a strain in which the gene of a predicted cellulose synthase was inactivated also indicates a role of cellulose in surface attachment. We propose that the mechanism of attachment via cellulose-anchored amyloidal fimbriae is widespread in bacteria and may function in initiation of infection and in formation of biofilms. PMID- 19682262 TI - Sequestration of essential proteins causes prion associated toxicity in yeast. AB - Prions are infectious, aggregated proteins that cause diseases in mammals but are not normally toxic in fungi. Excess Sup35p, an essential yeast protein that can exist as the [PSI(+)] prion, inhibits growth of [PSI(+)] but not [psi(-)] cells. This toxicity is rescued by expressing the Sup35Cp domain of Sup35p, which is sufficient for cell viability but not prion propagation. We now show that rescue requires Sup35Cp levels to be proportional to Sup35p overexpression. Overexpression of Sup35p appeared to cause pre-existing [PSI(+)] aggregates to coalesce into larger aggregates, but these were not toxic per se because they formed even when Sup35Cp rescued growth. Overexpression of Sup45p, but not other tested essential Sup35p binding partners, caused rescue. Sup45-GFPp formed puncta that colocalized with large [PSI(+)] Sup35-RFPp aggregates in cells overexpressing Sup35p, and the frequency of the Sup45-GFPp puncta was reduced by rescuing levels of Sup35Cp. In contrast, [PSI(+)] toxicity caused by a high excess of the Sup35p prion domain (Sup35NMp) was rescued by a single copy of Sup35Cp, was not rescued by Sup45p overexpression and was not associated with the appearance of Sup45-GFPp puncta. This suggests [PSI(+)] toxicity caused by excess Sup35p verses Sup35NMp is, respectively, through sequestration/inactivation of Sup45p verses Sup35p. PMID- 19682263 TI - Haem-delivery proteins in cytochrome c maturation System II. AB - Cytochromes of the c-type function on the outer side of the cytoplasmic membrane in bacteria where they also are assembled from apo-cytochrome polypeptide and haem. Two distinctly different systems for cytochrome c maturation are found in bacteria. System I present in Escherichia coli has eight to nine different Ccm proteins. System II is found in Bacillus subtilis and comprises four proteins: CcdA, ResA, ResB and ResC. ResB and ResC are poorly understood polytopic membrane proteins required for cytochrome c synthesis. We have analysed these two B. subtilis proteins produced in E. coli and in the native organism. ResB is shown to bind protohaem IX and haem is found covalently bound to residue Cys-138. Results in B. subtilis suggest that also ResC can bind haem. Our results complement recent findings made with Helicobacter CcsBA supporting the hypothesis that ResBC as a complex translocates haem by attaching it to ResB on the cytoplasmic side of the membrane and then transferring it to an extra-cytoplasmic location in ResC, from where it is made available to the apo-cytochromes. PMID- 19682264 TI - Global position analysis of the Pseudomonas aeruginosa quorum-sensing transcription factor LasR. AB - In Pseudomonas aeruginosa quorum sensing (QS), the transcriptional regulator LasR controls the expression of more than 300 genes. Several of these genes are activated indirectly via a second, subordinate QS regulator, RhlR. Conserved sequence elements upstream of individual other genes have been shown to bind LasR in vitro. To comprehensively identify all regions that are bound by LasR in vivo, we employed chromatin immunoprecipitation in conjunction with microarray analysis. We identified 35 putative promoter regions that direct the expression of up to 74 genes. In vitro DNA binding studies allowed us to distinguish between cooperative and non-cooperative LasR binding sites, and allowed us to build consensus sequences according to the mode of binding. Five promoter regions were not previously recognized as QS-controlled. Two of the associated transcript units encode proteins involved in the cold-shock response and in Psl exopolysaccharide synthesis respectively. The LasR regulon includes seven genes encoding transcriptional regulators, while secreted factors and secretion machinery are the most over-represented functional categories overall. This supports the notion that the core function of LasR is to co-ordinate the production of extracellular factors, although many of its effects on global gene expression are likely mediated indirectly by regulatory genes under its control. PMID- 19682266 TI - Switching off small RNA regulation with trap-mRNA. AB - Small non-coding regulatory RNAs in bacteria have been shown predominantly to be tightly regulated at the level of transcription initiation, and sRNAs that function by an antisense mechanism on trans-encoded target mRNAs have been shown or predicted to act stoichiometrically. Here we show that MicM, which silences the expression of an outer membrane protein, YbfM under most growth conditions, does not become destabilized by target mRNA overexpression, indicating that the small RNA regulator acts catalytically. Furthermore, our regulatory studies suggested that control of micM expression is unlikely to operate at the level of transcription initiation. By employing a highly sensitive genetic screen we uncovered a novel RNA-based regulatory principle in which induction of a trap mRNA leads to selective degradation of a small regulatory RNA molecule, thereby abolishing the sRNA-based silencing of its cognate target mRNA. In the present case, antisense regulation by chb mRNA of the antisense regulator MicM by an extended complementary sequence element, results in induction of ybfM mRNA translation. This type of regulation is reminiscent of the regulation of microRNA activity through target mimicry that occurs in plants. PMID- 19682265 TI - Targeted sigma factor turnover inserts negative control into a positive feedback loop. AB - Since their classification as members of the sigma(70) superfamily, Group IV alternative sigma factors have been found to control gene expression in response to diverse environmental or stress signals. Activity of the Streptomyces coelicolor Group IV family member, sigma(R) (SigR), is increased by changes in the oxidation-reduction state of cytoplasmic disulphide bonds. Once released by its cognate anti-sigma factor RsrA, sigma(R) activates expression of gene products that help cells reduce cytoplasmic disulphide bonds. In this issue of Molecular Microbiology, Kim and co-workers provide new insights into positive and negative control of sigma(R) activity. The authors show that a transcript derived from the inducible sigma(R)-dependent sigRrsrA p2 promoter operon encodes a sigma(R) protein of a higher molecular weight (termed sigma(R')) than is found in uninduced cells. One major difference between sigma(R') and the smaller sigma(R) protein found in uninduced cells is the rapid proteolysis of sigma(R') by the ClpP1/P2 protease system. The genes for the ClpP1/ClpP2 protease subunits are themselves members of the sigma(R) regulon. The newly identified positive (sigma(R') synthesis) and negative control (selective sigma(R') turnover) aspects of this circuit are either found or predicted to exist in other related Group IV sigma factor family members. PMID- 19682268 TI - Quantification of heterophoria and phoria adaptation using an automated objective system compared to clinical methods. AB - PURPOSE: To develop a system with which to quantify objectively the heterophoria to be measured throughout eye movement experiments. This study compared precision, resolution and accuracy characteristics of a limbus eye movement tracking system to the alternate cover test and the Maddox rod. METHODS: Precision testing was performed using each technique with binocularly-normal subjects at different test sessions. Resolution was measured and compared between the three systems. The heterophoria or phoria was measured from 40 cm/16 inches objectively with the limbus eye movement system and the Maddox rod during a sustained convergence-induced phoria adaptation experiment using physical and haploscope visual targets to compare accuracy. RESULTS: Precision testing quantified that the responses objectively recorded using the limbus tracking system exhibited similar standard deviations to the Maddox rod and the alternate cover test techniques. The limbus tracking method has the ability to quantify the response decay to phoria and has better resolution when compared to the clinical methods. When physical targets were used to induce phoria adaptation, the correlation was significant between the limbus eye movement tracking system and the Maddox rod after near and far adaptation, where the Maddox rod measurements were slightly more esophoric compared to those measured with the limbus tracking system. CONCLUSION: This objective limbus tracking system offers a better means by which to study the phoria and its adaptation throughout an eye movement experiment. The broader aim of this research is to establish a tool that will further the basic science of oculomotor control and binocular dysfunctions. PMID- 19682270 TI - Editorial regarding "relationship between pain and neuropathic symptoms in chronic musculoskeletal pain" by Dr. Giske et al. PMID- 19682267 TI - Mucosal mast cells mediate motor response induced by chronic oral exposure to ovalbumin in the rat gastrointestinal tract. AB - We previously demonstrated that oral chronic exposure to ovalbumin (OVA) causes intestinal hypermotility in Sprague-Dawley rats. In this study, the objective was to determine the mechanism of action of OVA and the role of mucosal mast cells in the regulation of motor activity in this model. Rats were orally exposed to OVA during 6 weeks. Intestinal mucosal mast cells (IMMCs) were counted and rat mast cell protease II (RMCPII) measured in duodenum, jejunum, ileum and colon. Anti OVA IgE, IgG, and IL-4 were measured in serum. Eosinophils and IgE(+) cells were counted in jejunum. In an additional study rats were treated with the mast cell stabilizer ketotifen and mast cell number, RMCPII concentration and motor activity in vitro were evaluated. OVA exposed rats showed an increase in mucosal mast cell number and in RMCPII content in small intestine and colon. However, variables of a Th(2) type response were not affected by exposure to OVA: (i) neither OVA specific IgE nor IgG were found; (ii) IL-4 did not increase and, (iii) the number of eosinophils and IgE(+) cells was identical in the exposed and unexposed groups. These results brought us to hypothesize a possible non-Ig mediated action of OVA on mast cells. Ketotifen significantly diminished the response to OVA: Ketotifen reduced the number of mast cells and the RMCPII content and blocked increased intestinal contractility. In addition ketotifen modified motor response in both OVA exposed and unexposed animals giving evidence of the importance of mast cells in intestine motor activity driving. PMID- 19682271 TI - AAPM and health care reform: your voice is needed. PMID- 19682272 TI - Depressive symptoms predict the subsequent risk of bodily pain in dialysis patients: Japan Dialysis Outcomes and Practice Patterns Study. AB - BACKGROUND: Bodily pain and psychiatric distress are common symptoms in patients with dialysis. However, the temporal relationships have not yet been investigated. Objective. To evaluate the longitudinal association between depressive symptoms and subsequent risk of developing severe bodily pain in dialysis patients. Design. Prospective cohort study. METHODS: We assessed bodily pain using a self-reported questionnaire and depressive symptoms using scores from the short version of Center for Epidemiological Studies Depression Screening Index (CES-D) from 531 participants showing no/mild bodily pain at baseline, based on the Japan Dialysis Outcomes and Practice Patterns Study, a cohort study of hemodialysis patients. To evaluate the relationship between depressive symptoms and development of severe bodily pain, multivariable logistic regression analysis was performed. RESULTS: The 531 patients had a mean age of 57.9 years, 61.4% were male, and 33.1% had depressive symptoms. Logistic regression analysis revealed that depressive symptoms at baseline were significantly associated with higher odds of developing severe bodily pain during a 0.5- to 2.5-year follow-up period (adjusted odds ratio [AOR] = 2.13, 95% confidence interval [CI]: 1.36 3.33, P = 0.001). Further, patients with higher CES-D scores were likely to develop severe bodily pain (AOR = 1.09, 95% CI: 1.04-1.15, P = 0.001). CONCLUSIONS: Results of this study suggest that depressive symptoms measured by CES-D predict the future risk of developing severe bodily pain in dialysis patients. PMID- 19682274 TI - Efficacy of a metered-dose 8% lidocaine pump spray for patients with post herpetic neuralgia. AB - OBJECTIVE: Topical lidocaine patch is effective in the treatment of post-herpetic neuralgia (PHN), but not suited for paroxysmal pain because of the long latency of analgesia. Here, we examined the efficacy of 8% lidocaine pump spray (Xylocaine pump spray, XPS) for PHN. DESIGN: Twenty-four patients with PHN were recruited into a randomized, double-blind, placebo-controlled, crossover study (study 1), and 100 patients with PHN were recruited into an open-labeled study (study 2). In study 1, patients were randomized to receive either XPS or saline placebo pump spray (PPS) applied to the painful skin areas. Following a 7-day period, patients were crossed over to receive the alternative treatment. In study 2, XPS was prescribed for patients who were advised to use the spray anytime, with a 2-hour gap between applications, for 2 weeks. The pain was assessed with a visual analogue scale (VAS). Details of use were noted in the diary. RESULTS: In study 1, greater decreases in VAS of persistent pain followed application of XPS (baseline: 6.1 +/- 1.7 cm, 15-minute post-spray: 2.3 +/- 2.5 cm, mean +/- SD) than with PPS (6.1 +/- 1.7 cm, 5.7 +/- 1.6 cm, [P < 0.01]). The effect persisted for a median of 4.5 hours (range, 2 to 24 hours) after application. In study 2, 13 of 100 patients discontinued the treatment because of mild local side effects or insufficient effect. In the remaining 87 patients, XPS maintained significant pain relief relative to baseline throughout the 2-week period. Satisfaction with the therapy was reported by 79% of patients. CONCLUSIONS: In both studies, XPS provided a significant improvement in PHN due to its prompt analgesia, lack of systemic side effects, and convenience of use. PMID- 19682273 TI - Prevention and reversal of morphine tolerance by the analgesic neuroactive steroid alphadolone. AB - OBJECTIVE: Alphadolone is a neuroactive steroid that causes antinociception in rats and analgesia in humans by interaction with spinal cord GABA(A) receptors. This study investigated whether alphadolone could affect morphine tolerance. METHODS: Morphine tolerance was induced in rats with subcutaneous sustained release morphine emulsion (M-SR; 125 mg/kg/day). Tolerance was assessed by a blinded observer using tail flick latency (TFL) response to intraperitoneal (ip) injection of immediate release morphine (M-IR 6.25 mg/kg). Fifty-five rats given M-SR were divided into three groups: group A received 1.0 mL subcutaneous emulsion containing vehicle; groups B and C had emulsions injected subcutaneously at the same time as the M-SR (B-250 mg/kg alphadolone; C-alphaxalone 80 mg/kg). RESULTS: Tail flick latency responses (percentage of maximum possible effect [% MPE]) to M-IR were reduced from 89.6 +/- 2.5 pretreatment to 20.3 +/- 4.8 after M SR treatment (mean +/- SEM; P < 0.001, one-way anova). Coadministration of alphadolone emulsion with the M-SR caused no sedation and prevented the occurrence of morphine tolerance. TFL responses to M-IR (6.25 mg/kg) given to morphine tolerant rats were 29 +/- 8% MPE whereas the TFL was 78.6 +/- 9.8% MPE when immediate release alphadolone (10 mg/kg ip) was injected at the same time as M-IR to tolerant rats (P < 0.001 one-way anova). Alphaxalone treatment caused sedation and no effects on morphine tolerance. CONCLUSIONS: We conclude that the alphadolone can prevent morphine tolerance and it also restores normal morphine antinociception in rats with established morphine tolerance. The lack of sedation suggests clinical utility in human pain states requiring morphine. PMID- 19682275 TI - Care in the interpretation of the SAFE algorithm. PMID- 19682277 TI - Comparability of a hand-held nitric oxide analyser with online and offline chemiluminescence-based nitric oxide measurement. AB - Practicability is crucial for successful implementation of fractional exhaled nitric oxide (FeNO) measurement into asthma management. The study aimed at comparing a conventional chemiluminescence NO analyser (EcoMedics) with a hand held device (NIOX MINO) and offline FeNO measurement using a commercially available system in an unselected cohort of children aged 6-16 yr. A secondary objective was to confirm FeNO stability over time in 15 samples from adult volunteers obtained using the offline system. Sixty-six children (mean +/- s.d. age 11.8 +/- 3.0 yr) underwent single breath FeNO measurement in triplets with each device. Offline collected FeNO was measured after offline breath collection into a Mylar balloon and subsequent analysis using the chemiluminescence NO analyser. Variability and between-method agreement were assessed, and stability over time within the Mylar balloons was tested by repeated hourly measurements. FeNO levels ranged from 2 to 113 p.p.b. Intra-class correlation was excellent (r = 0.98, p < 0.001 for each pair). Bland-Altman plots and back-transformation of logarithmic mean differences revealed fair agreement between methods. Stability over time was confirmed over 10 h both at room temperature and when stored under cooling conditions. FeNO values obtained using the chemiluminescence NO analyser, the portable NIOX MINO system and the offline collection technique show between method agreement within clinically acceptable range. PMID- 19682279 TI - The alternative oxidase family of Vitis vinifera reveals an attractive model to study the importance of genomic design. AB - 'Genomic design' refers to the structural organization of gene sequences. Recently, the role of intron sequences for gene regulation is being better understood. Further, introns possess high rates of polymorphism that are considered as the major source for speciation. In molecular breeding, the length of gene-specific introns is recognized as a tool to discriminate genotypes with diverse traits of agronomic interest. 'Economy selection' and 'time-economy selection' have been proposed as models for explaining why highly expressed genes typically contain small introns. However, in contrast to these theories, plant specific selection reveals that highly expressed genes contain introns that are large. In the presented research, 'wet'Aox gene identification from grapevine is advanced by a bioinformatics approach to study the species-specific organization of Aox gene structures in relation to available expressed sequence tag (EST) data. Two Aox1 and one Aox2 gene sequences have been identified in Vitis vinifera using grapevine cultivars from Portugal and Germany. Searching the complete genome sequence data of two grapevine cultivars confirmed that V. vinifera alternative oxidase (Aox) is encoded by a small multigene family composed of Aox1a, Aox1b and Aox2. An analysis of EST distribution revealed high expression of the VvAox2 gene. A relationship between the atypical long primary transcript of VvAox2 (in comparison to other plant Aox genes) and its expression level is suggested. V. vinifera Aox genes contain four exons interrupted by three introns except for Aox1a which contains an additional intron in the 3'-UTR. The lengths of primary Aox transcripts were estimated for each gene in two V. vinifera varieties: PN40024 and Pinot Noir. In both varieties, Aox1a and Aox1b contained small introns that corresponded to primary transcript lengths ranging from 1501 to 1810 bp. The Aox2 of PN40024 (12 329 bp) was longer than that from Pinot Noir (7279 bp) because of selection against a transposable-element insertion that is 5028 bp in size. An EST database basic local alignment search tool (BLAST) search of GenBank revealed the following ESTs percentages for each gene: Aox1a (26.2%), Aox1b (11.9%) and Aox2 (61.9%). Aox1a was expressed in fruits and roots, Aox1b expression was confined to flowers and Aox2 was ubiquitously expressed. These data for V. vinifera show that atypically long Aox intron lengths are related to high levels of gene expression. Furthermore, it is shown for the first time that two grapevine cultivars can be distinguished by Aox intron length polymorphism. PMID- 19682278 TI - Induction of IL-10+ CD4+ CD25+ regulatory T cells with decreased NF-kappaB expression during immunotherapy. AB - MyD88 is a major toll-like receptor (TLR) adaptor to activate NF-kappaB, which acts as a mater switch for allergic inflammation disease. Sterile hust dust extracts have been reported with TLR-dependent immunostimulatory activities. The aim of this study was to evaluate whether Dermatophagoides pteronyssinus (Der p) immunotherapy may increase IL-10+ CD4+ CD25+ T cells with modulating MyD88 signaling proteins, to decrease NF-kappaB expression. Peripheral blood mononuclear cells were isolated from patients before and after 1 yr of Der p immunotherapy, and also from matched control subjects. After 2 days of Der p-2 stimulation, intracellular IL-10 and Foxp3 expression of CD4(+) CD25(+) T cells were measured by flow-cytometry. The expression of IL-1 receptor-associated kinase (IRAK)-1 in cytoplasm and IFN-regulator factor-3 (IRF-3) with NF kappaB/p65 in nuclei was determined by Western-blot analysis. Patients undergoing immunotherapy produced more soluble CD14, IL-10, and TGF-beta that correlated with FEV(1) improvement (p < 0.05). In the immunotherapy group, the number of Foxp3+ CD4+ Treg cells increased more than the baseline status (25.06 +/- 4.19 vs. 16.08 +/- 3.54, p < 0.05). Additionally, increased IL-10 production with decreased IRAK-1 and NF-kappaB/p65 nuclear translocation was observed in sorted purified Treg cells. IL-10(+) CD4(+) CD25(+) Treg cells may respond to Der p-2 and down-regulate NF-kappaB/p65 expression to maintain immune tolerance during immunotherapy. PMID- 19682280 TI - Identification of direct transcriptional targets of (V600E)BRAF/MEK signalling in melanoma. AB - Oncogenic mutations in BRAF are common in melanoma and drive constitutive activation of the MEK/ERK pathway. To elucidate the transcriptional events downstream of (V600E)BRAF/MEK signalling we performed gene expression profiling of A375 melanoma cells treated with potent and selective inhibitors of (V600E)BRAF and MEK (PLX4720 and PD184352 respectively). Using a stringent Bayesian approach, we identified 69 transcripts that appear to be direct transcriptional targets of this pathway and whose expression changed after 6 h of pathway inhibition. We also identified several additional genes whose expression changed after 24 h of pathway inhibition and which are likely to be indirect transcriptional targets of the pathway. Several of these were confirmed by demonstrating their expression to be similarly regulated when BRAF was depleted using RNA interference, and by using qRT-PCR in other BRAF mutated melanoma lines. Many of these genes are transcription factors and feedback inhibitors of the ERK pathway and are also regulated by MEK signalling in NRAS mutant cells. This study provides a basis for understanding the molecular processes that are regulated by (V600E)BRAF/MEK signalling in melanoma cells. PMID- 19682282 TI - Tailoring intensified insulin therapy in children. PMID- 19682281 TI - Stem cell factor rescues tyrosinase expression and pigmentation in discreet anatomic locations in albino mice. AB - The K14-SCF transgenic murine model of variant pigmentation is based on epidermal expression of stem cell factor (SCF) on the C57BL/6J background. In this system, constitutive expression of SCF by epidermal keratinocytes results in retention of melanocytes in the interfollicular basal layer and pigmentation of the epidermis itself. Here, we extend this animal model by developing a compound mutant transgenic amelanotic animal defective at both the melanocortin 1 receptor (Mc1r) and tyrosinase (Tyr) loci. In the presence of K14-Scf, tyrosinase-mutant animals (previously thought incapable of synthesizing melanin) exhibited progressive robust epidermal pigmentation with age in the ears and tails. Furthermore, K14 SCF Tyr(c2j/c2j) animals demonstrated tyrosinase expression and enzymatic activity, suggesting that the c2j Tyr defect can be rescued in part by SCF in the ears and tail. Lastly, UV sensitivity of K14-Scf congenic animals depended mainly on the amount of eumelanin present in the skin. These findings suggest that c-kit signaling can overcome the c2j Tyr mutation in the ears and tails of aging animals and that UV resistance depends on accumulation of epidermal eumelanin. PMID- 19682283 TI - A rice tryptophan deficient dwarf mutant, tdd1, contains a reduced level of indole acetic acid and develops abnormal flowers and organless embryos. AB - Indole-3-acetic acid (IAA) plays a critical role in many aspects of plant growth and development; however, complete pathways of biosynthesis, localization and many aspects of functions of IAA in rice remain unclear. Here, we report the analysis of a rice tryptophan- (Trp-) and IAA-deficient mutant, tryptophan deficient dwarf1 (tdd1), which is embryonic lethal because of a failure to develop most organs during embryogenesis. Regenerated tdd1 plants showed pleiotropic phenotypes: dwarfing, narrow leaves, short roots and abnormal flowers. TDD1 encodes a protein homologous to anthranilate synthase beta-subunit, which catalyses the first step of the Trp biosynthesis pathway and functions upstream of Trp-dependent IAA biosynthesis. TDD1-uidA and DR5-uidA expression overlapped at many sites in WT plants but was lacking in tdd1, indicating that TDD1 is involved in auxin biosynthesis. Both Trp and IAA levels in flowers and embryos were much lower in tdd1 than in wild type (WT). Trp feeding completely rescued the mutant phenotypes and moderate expression of OsYUCCA1, which encodes a key enzyme in Trp-dependent IAA biosynthesis, also rescued plant height and root length, indicating that the abnormal phenotypes of tdd1 are caused predominantly by Trp and IAA deficiency. In tdd1 embryos, the expression patterns of OSH1 and OsSCR, which mark the presumptive apical region and the L2 layer, respectively, are identical to those in WT, suggesting a possibility either that different IAA levels are required for basic pattern formation than for organ formation or that an orthologous gene compensates for TDD1 deficiency during pattern formation. PMID- 19682284 TI - Single-nucleotide and long-patch base excision repair of DNA damage in plants. AB - Base excision repair (BER) is a critical pathway in cellular defense against endogenous or exogenous DNA damage. This elaborate multistep process is initiated by DNA glycosylases that excise the damaged base, and continues through the concerted action of additional proteins that finally restore DNA to the unmodified state. BER has been subject to detailed biochemical analysis in bacteria, yeast and animals, mainly through in vitro reproduction of the entire repair reaction in cell-free extracts. However, an understanding of this repair pathway in plants has consistently lagged behind. We report the extension of BER biochemical analysis to plants, using Arabidopsis cell extracts to monitor repair of DNA base damage in vitro. We have used this system to demonstrate that Arabidopsis cell extracts contain the enzymatic machinery required to completely repair ubiquitous DNA lesions, such as uracil and abasic (AP) sites. Our results reveal that AP sites generated after uracil excision are processed both by AP endonucleases and AP lyases, generating either 5'- or 3'-blocked ends, respectively. We have also found that gap filling and ligation may proceed either through insertion of just one nucleotide (short-patch BER) or several nucleotides (long-patch BER). This experimental system should prove useful in the biochemical and genetic dissection of BER in plants, and contribute to provide a broader picture of the evolution and biological relevance of DNA repair pathways. PMID- 19682285 TI - Genotype and time of day shape the Populus drought response. AB - As exposure to episodic drought can impinge significantly on forest health and the establishment of productive tree plantations, there is great interest in understanding the mechanisms of drought response in trees. The ecologically dominant and economically important genus Populus, with its sequenced genome, provides an ideal opportunity to examine transcriptome level changes in trees in response to a drought stimulus. The transcriptome level drought response of two commercially important Populus clones (P. deltoides x P. nigra, DN34, and P. nigra x P. maximowiczii, NM6) was characterized over a diurnal period using a 4 x 2 x 2 complete randomized factorial anova experimental design (four time points, two genotypes and two treatment conditions), using Affymetrix Poplar GeneChip microarrays. Notably, the specific genes that exhibited changes in transcript abundance in response to drought differed between the genotypes and/or the time of day that they exhibited their greatest differences. This study emphasizes the fact that it is not possible to draw simple, generalized conclusions about the drought response of the genus Populus on the basis of one species, nor on the basis of results collected at a single time point. The data derived from our studies provide insights into the variety of genetic mechanisms underpinning the Populus drought response, and provide candidates for future experiments aimed at understanding this response across this economically and ecologically important genus. PMID- 19682286 TI - The Arabidopsis SUMO E3 ligase AtMMS21, a homologue of NSE2/MMS21, regulates cell proliferation in the root. AB - hMMS21 is a SUMO E3 ligase required for the prevention of DNA damage-induced apoptosis, and acts by facilitating DNA repair in human cells. The Arabidopsis genome contains a putative MMS21 homologue capable of interacting with the SUMO E2 conjugating enzyme AtSCE1a, as indicated by a yeast two-hybrid screen and bimolecular fluorescence complementation experiments. In vitro and in vivo data demonstrated that AtMMS21 was a SUMO E3 ligase. We identified the Arabidopsis AtMMS21 null T-DNA insertion mutant mms21-1, which had a short-root phenotype, and affected cell proliferation in the apical root meristem, as indicated by impaired expression of the cell division marker CYCB1:GUS in mms21-1 roots. The mms21-1 roots had reduced responses to exogenous cytokinins, and decreased expression of the cytokinin-induced genes ARR3, ARR4, ARR5 and ARR7, compared with the wild type. Thus, our findings suggest that the AtMMS21 gene is involved in root development via cell-cycle regulation and cytokinin signalling. PMID- 19682287 TI - FATTY ACID DESATURASE4 of Arabidopsis encodes a protein distinct from characterized fatty acid desaturases. AB - Polar membrane glycerolipids occur in a mixture of molecular species defined by a polar head group and characteristic acyl groups esterified to a glycerol backbone. A molecular species of phosphatidylglycerol specific to chloroplasts of plants carries a Delta(3-trans) hexadecenoic acid in the sn-2 position of its core glyceryl moiety. The fad4-1 mutant of Arabidopsis thaliana missing this particular phosphatidylglycerol molecular species lacks the necessary fatty acid desaturase, or a component thereof. The overwhelming majority of acyl groups associated with membrane lipids in plants contains double bonds with a cis configuration. However, FAD4 is unusual because it is involved in the formation of a trans double bond introduced close to the carboxyl group of palmitic acid, which is specifically esterified to the sn-2 glyceryl carbon of phosphatidylglycerol. As a first step towards the analysis of this unusual desaturase reaction, the FAD4 gene was identified by mapping of the FAD4 locus and coexpression analysis with known lipid genes. FAD4 encodes a predicted integral membrane protein that appears to be unrelated to classic membrane bound fatty acid desaturases based on overall sequence conservation. However, the FAD4 protein contains two histidine motifs resembling those of metalloproteins such as fatty acid desaturases. FAD4 is targeted to the plastid. Overexpression of the cDNA in transgenic Arabidopsis led to increased accumulation of the Delta(3 trans) hexadecanoyl group in phosphatidylglycerol relative to wild type. Taken together these results are consistent with the hypothesis that FAD4 is the founding member of a novel class of fatty acid desaturases. PMID- 19682288 TI - Proteomics reveals the overlapping roles of hydrogen peroxide and nitric oxide in the acclimation of citrus plants to salinity. AB - Hydrogen peroxide (H(2)O(2)) and nitric oxide (*NO) are key reactive species in signal transduction pathways leading to activation of plant defense against biotic or abiotic stress. Here, we investigated the effect of pre-treating citrus plants (Citrus aurantium L.) with either of these two molecules on plant acclimation to salinity and show that both pre-treatments strongly reduced the detrimental phenotypical and physiological effects accompanying this stress. A proteomic analysis disclosed 85 leaf proteins that underwent significant quantitative variations in plants directly exposed to salt stress. A large part of these changes was not observed with salt-stressed plants pre-treated with either H(2)O(2) or sodium nitroprusside (SNP; a *NO-releasing chemical). We also identified several proteins undergoing changes either in their oxidation (carbonylation; 40 proteins) and/or S-nitrosylation (49 proteins) status in response to salinity stress. Both H(2)O(2) and SNP pre-treatments before salinity stress alleviated salinity-induced protein carbonylation and shifted the accumulation levels of leaf S-nitrosylated proteins to those of unstressed control plants. Altogether, the results indicate an overlap between H(2)O(2)- and *NO-signaling pathways in acclimation to salinity and suggest that the oxidation and S-nitrosylation patterns of leaf proteins are specific molecular signatures of citrus plant vigour under stressful conditions. PMID- 19682289 TI - Tethering of ferredoxin:NADP+ oxidoreductase to thylakoid membranes is mediated by novel chloroplast protein TROL. AB - Working in tandem, two photosystems in the chloroplast thylakoid membranes produce a linear electron flow from H(2)O to NADP(+). Final electron transfer from ferredoxin to NADP(+) is accomplished by a flavoenzyme ferredoxin:NADP(+) oxidoreductase (FNR). Here we describe TROL (thylakoid rhodanese-like protein), a nuclear-encoded component of thylakoid membranes that is required for tethering of FNR and sustaining efficient linear electron flow (LEF) in vascular plants. TROL consists of two distinct modules; a centrally positioned rhodanese-like domain and a C-terminal hydrophobic FNR binding region. Analysis of Arabidopsis mutant lines indicates that, in the absence of TROL, relative electron transport rates at high-light intensities are severely lowered accompanied with significant increase in non-photochemical quenching (NPQ). Thus, TROL might represent a missing thylakoid membrane docking site for a complex between FNR, ferredoxin and NADP(+). Such association might be necessary for maintaining photosynthetic redox poise and enhancement of the NPQ. PMID- 19682290 TI - Electrophysiological characterization of pathways for K(+) uptake into growing and non-growing leaf cells of barley. AB - Potassium is a major osmolyte used by plant cells. The accumulation rates of K(+) in cells may limit the rate of expansion. In the present study, we investigated the involvement of ion channels in K(+) uptake using patch clamp technique. Ion currents were quantified in protoplasts of the elongation and emerged blade zone of the developing leaf 3 of barley (Hordeum vulgare L.). A time-dependent inward rectifying K(+)-selective current was observed almost exclusively in elongation zone protoplasts. The current showed characteristics typical of Shaker-type channels. Instantaneous inward current was highest in the epidermis of the emerged blade and selective for Na(+) over K(+). Selectivity disappeared, and currents decreased or remained the same, depending on tissue, in response to salt treatment. Net accumulation rates of K(+) in cells calculated from patch clamp current-voltage curves exceeded rates calculated from membrane potential and K(+) concentrations of cells measured in planta by factor 2.5-2.7 at physiological apoplastic K(+) concentrations (10-100 mm). It is concluded that under these conditions, K(+) accumulation in growing barley leaf cells is not limited by transport properties of cells. Under saline conditions, down-regulation of voltage-independent channels may reduce the capacity for growth-related K(+) accumulation. PMID- 19682291 TI - Potassium channels in barley: cloning, functional characterization and expression analyses in relation to leaf growth and development. AB - It is not known how the uptake and retention of the key osmolyte K(+) in cells are mediated in growing leaf tissue. In the present study on the growing leaf 3 of barley, we have cloned the full-length coding sequence of three genes which encode putative K(+) channels (HvAKT1, HvAKT2, HvKCO1/HvTPK1), and of one gene which encodes a putative K(+) transporter (HvHAK4). The functionality of the gene products of HvAKT1 and HvAKT2 was tested through expression in Xenopus laevis oocytes. Both are inward-rectifying K(+) channels which are inhibited by Cs(+). Function of HvAKT1 in oocytes requires co-expression of a calcineurin-interacting protein kinase (AtCIPK23) and a calcineurin B-like protein (AtCBL9) from Arabidopsis, showing cross-species complementation of function. In planta, HvAKT1 is expressed primarily in roots, but is also expressed in leaf tissue. HvAKT2 is expressed particularly in leaf tissue, and HvHAK4 is expressed particularly in growing leaf tissue. Within leaves, HvAKT1 and HvAKT2 are expressed predominantly in mesophyll. Expression of genes changes little in response to low external K(+) or salinity, despite major changes in K(+) concentrations and osmolality of cells. Possible contributions of HvAKT1, HvAKT2, HvKCO1 and HvHAK4 to regulation of K(+) relations of growing barley leaf cells are discussed. PMID- 19682292 TI - Increased nitrogen-use efficiency in transgenic rice plants over-expressing a nitrogen-responsive early nodulin gene identified from rice expression profiling. AB - Development of genetic varieties with improved nitrogen-use efficiency (NUE) is essential for sustainable agriculture. In this study, we developed a growth system for rice wherein N was the growth-limiting factor, and identified N responsive genes by a whole genome transcriptional profiling approach. Some genes were selected to test their functionality in NUE by a transgenic approach. One such example with positive effects on NUE is an early nodulin gene OsENOD93-1. This OsENOD93-1 gene responded significantly to both N induction and N reduction. Transgenic rice plants over-expressing the OsENOD93-1 gene had increased shoot dry biomass and seed yield. This OsENOD93-1 gene was expressed at high levels in roots of wild-type (WT) plants, and its protein product was localized in mitochondria. Transgenic plants accumulated higher concentrations of total amino acids and total N in roots. A higher concentration of amino acids in xylem sap was detected in transgenic plants, especially under N stress. In situ hybridization revealed that OsENOD93-1 is expressed in vascular bundles, as well as in epidermis and endodermis. This work demonstrates that transcriptional profiling, coupled with a transgenic validation approach, is an effective strategy for gene discovery. The knowledge gained from this study could be applied to other important crops. PMID- 19682293 TI - Plasticity in maximum stomatal conductance constrained by negative correlation between stomatal size and density: an analysis using Eucalyptus globulus. AB - Maximum stomatal conductance to water vapour and CO2 (gwmax, gcmax, respectively), which are set at the time of leaf maturity, are determined predominantly by stomatal size (S) and density (D). In theory, many combinations of S and D yield the same gwmax and gcmax, so there is no inherent correlation between S and D, or between S, D and maximum stomatal conductance. However, using basic equations for gas diffusion through stomata of different sizes, we show that a negative correlation between S and D offers several advantages, including plasticity in gwmax and gcmax with minimal change in epidermal area allocation to stomata. Examination of the relationship between S and D in Eucalyptus globulus seedlings and coppice shoots growing in the field under high and low rainfall revealed a strong negative relationship between S and D, whereby S decreased with increasing D according to a negative power function. The results provide evidence that plasticity in maximum stomatal conductance may be constrained by a negative S versus D relationship, with higher maximum stomatal conductance characterized by smaller S and higher D, and a tendency to minimize change in epidermal space allocation to stomata as S and D vary. PMID- 19682294 TI - Recombineering and stable integration of the Pseudomonas syringae pv. syringae 61 hrp/hrc cluster into the genome of the soil bacterium Pseudomonas fluorescens Pf0 1. AB - Many Gram-negative bacteria use a type III secretion system (T3SS) to establish associations with their hosts. The T3SS is a conduit for direct injection of type III effector proteins into host cells, where they manipulate the host for the benefit of the infecting bacterium. For plant-associated pathogens, the variations in number and amino acid sequences of type-III effectors, as well as their functional redundancy, make studying type-III effectors challenging. To mitigate this challenge, we developed a stable delivery system for individual or defined sets of type-III effectors into plant cells. We used recombineering and Tn5-mediated transposition to clone and stably integrate, respectively, the complete hrp/hrc region from Pseudomonas syringae pv. syringae 61 into the genome of the soil bacterium Pseudomonas fluorescens Pf0-1. We describe our development of Effector-to-Host Analyzer (EtHAn), and demonstrate its utility for studying effectors for their in planta functions. PMID- 19682295 TI - Tissue and cellular localization of individual beta-glycosidases using a substrate-specific sugar reducing assay. AB - Traditional methods to localize beta-glycosidase activity in tissue sections have been based on incubation with the general substrate 6-bromo-2-naphthyl-beta-d glucopyranoside. When hydrolysed in the presence of salt zinc compounds, 6-bromo 2-naphthyl-beta-d-glucopyranoside affords the formation of an insoluble coloured product. This technique does not distinguish between different beta-glycosidases present in the tissue. To be able to monitor the occurrence of individual beta glycosidases in different tissues and cell types, we have developed a versatile histochemical method that can be used for localization of any beta-glycosidase that upon incubation with its specific substrate releases a reducing sugar. Experimentally, the method is based on hydrolysis of the specific substrate followed by oxidation of the sugar released by a tetrazolium salt (2,3,5 triphenyltetrazolium chloride) that forms a red insoluble product when reduced. The applicability of the method was demonstrated by tissue and cellular localization of two beta-glucosidases, amygdalin hydrolase and prunasin hydrolase, in different tissues and cell types of almond. In those cases where the analysed tissue had a high content of reducing sugars, this resulted in strong staining of the background. This interfering staining of the background was avoided by prior incubation with sodium borohydride. The specificity of the devised method was demonstrated in a parallel localization study using a specific antibody towards prunasin hydrolase. PMID- 19682296 TI - Mutations in the cinnamate 4-hydroxylase gene impact metabolism, growth and development in Arabidopsis. AB - The initial reactions of the phenylpropanoid pathway convert phenylalanine to p coumaroyl CoA, a branch point metabolite from which many phenylpropanoids are made. Although the second enzyme of this pathway, cinnamic acid 4-hydroxylase (C4H), is well characterized, a mutant for the gene encoding this enzyme has not yet, to our knowledge, been identified, presumably because knock-out mutations in this gene would have severe phenotypes. This work describes the characterization of an allelic series of Arabidopsis reduced epidermal fluorescence 3 (ref3) mutants, each of which harbor mis-sense mutations in C4H (At2g30490). Heterologous expression of the mutant proteins in Escherichia coli yields enzymes that exhibit P420 spectra, indicative of mis-folded proteins, or have limited ability to bind substrate, indicating that the mutations we have identified affect protein stability and/or enzyme function. In agreement with the early position of C4H in phenylpropanoid metabolism, ref3 mutant plants accumulate decreased levels of several different classes of phenylpropanoid end-products, and exhibit reduced lignin deposition and altered lignin monomer content. Furthermore, these plants accumulate a novel hydroxycinnamic ester, cinnamoylmalate, which is not found in the wild type. The decreased C4H activity in ref3 also causes pleiotropic phenotypes, including dwarfism, male sterility and the development of swellings at branch junctions. Together, these observations indicate that C4H function is critical to the normal biochemistry and development of Arabidopsis. PMID- 19682297 TI - An F-box gene, CPR30, functions as a negative regulator of the defense response in Arabidopsis. AB - Arabidopsis gain-of-resistance mutants, which show HR-like lesion formation and SAR-like constitutive defense responses, were used well as tools to unravel the plant defense mechanisms. We have identified a novel mutant, designated constitutive expresser of PR genes 30 (cpr30), that exhibited dwarf morphology, constitutive resistance to the bacterial pathogen Pseudomonas syringae and the dramatic induction of defense-response gene expression. The cpr30-conferred growth defect morphology and defense responses are dependent on ENHANCED DISEASE SUSCEPTIBILITY 1 (EDS1), PHYTOALEXIN DEFICIENT 4 (PAD4), and NONRACE-SPECIFIC DISEASE RESISTANCE 1 (NDR1). Further studies demonstrated that salicylic acid (SA) could partially account for the cpr30-conferred constitutive PR1 gene expression, but not for the growth defect, and that the cpr30-conferred defense responses were NPR1 independent. We observed a widespread expression of CPR30 throughout the plant, and a localization of CPR30-GFP fusion protein in the cytoplasm and nucleus. As an F-box protein, CPR30 could interact with multiple Arabidopsis-SKP1-like (ASK) proteins in vivo. Co-localization of CPR30 and ASK1 or ASK2 was observed in Arabidopsis protoplasts. Based on these results, we conclude that CPR30, a novel negative regulator, regulates both SA-dependent and SA-independent defense signaling, most likely through the ubiquitin-proteasome pathway in Arabidopsis. PMID- 19682298 TI - Metabolic aspects of neonatal rat islet hypoxia tolerance. AB - Sensitivity of pancreatic islets to hypoxia is one of the most important of the obstacles responsible for their failure to survive within the recipients. The aim of this study was to compare the in vitro hypoxia tolerance of neonatal and adult rat islet cells and to study the glucose metabolism in these cells after exposure to hypoxia. Islet cells from both age categories were cultured in different hypoxic levels for 24 h and insulin secretion and some metabolites of glucose metabolism were analysed. Glucose-stimulated insulin secretion decreased dramatically in both cell preparations in response to the decrease in oxygen level. The reduction of insulin secretion was more detectable in adult cells and started at 5% O(2), while a significant reduction was obtained at 1% O(2) in neonatal cells. Moreover, basal insulin release of neonatal cells showed an adaptation to hypoxia after a 4-day culture in hypoxia. Intracellular pyruvate was higher in neonatal cells than in adult ones, while no difference in lactate level was observed between them. Similar results to that of pyruvate were observed for adenosine triphosphate (ATP) and the second messenger cyclic adenosine monophosphate (cAMP). The study reveals that neonatal rat islet cells are more hypoxia-tolerant than the adult ones. The most obvious metabolic observation was that both pyruvate and lactate were actively produced in neonatal cells, while adult cells depended mainly on lactate production as an end-product of glycolysis, indicating a more enhanced metabolic flexibility of neonatal cells to utilize the available oxygen and, at the same time, maintain metabolism anaerobically. PMID- 19682299 TI - Absence of residual Hodgkin's disease demonstrated by PET/CT in a deceased organ donor. AB - With the current limited availability of organs for transplantation, it is important to consider marginal donor candidates, including survivors of potentially curable malignancies such as lymphoma. The absence of refractory/recurrent residual disease at the time of brain death can be difficult to establish. Therefore, it is critical to have objective data to decide whether to proceed or not with organ procurement and transplantation. We report a unique situation in which (18)F-fluorodeoxyglucose positron emission tomography (PET) was used to rule out Hodgkin's lymphoma recurrence in a 33-year-old, heart beating, brain-dead, potential donor with a past history of Hodgkin's disease and a persistent mediastinal mass. PET showed no significant uptake in the mass, allowing organ donation and transplantation to occur. We present a new means of evaluating potential brain-dead donors with a past history of some lymphoma, whereby PET may help transplant physicians by optimizing donation safety while rationalizing the inclusion of marginal donors. PMID- 19682300 TI - The crystal structure of a xyloglucan-specific endo-beta-1,4-glucanase from Geotrichum sp. M128 xyloglucanase reveals a key amino acid residue for substrate specificity. AB - Geotrichum sp. M128 possesses two xyloglucan-specific glycoside hydrolases belonging to family 74, xyloglucan-specific endo-beta-1,4-glucanase (XEG) and oligoxyloglucan reducing-end-specific cellobiohydrolase (OXG-RCBH). Despite their similar amino acid sequences (48% identity), their modes of action and substrate specificities are distinct. XEG catalyzes the hydrolysis of xyloglucan polysaccharides in endo mode, while OXG-RCBH acts on xyloglucan oligosaccharides at the reducing end in exo mode. Here, we determined the crystal structure of XEG at 2.5 A resolution, and compared it to a previously determined structure of OXG RCBH. For the most part, the amino acid residues that interact with substrate are conserved between the two enzymes. However, there are notable differences at subsite positions -1 and +2. OXG-RCBH has a loop around the +2 site that blocks one end of the active site cleft, which accounts for its exo mode of action. In contrast, XEG lacks a corresponding loop at this site, thereby allowing binding to the middle of the main chain of the substrate. At the -1 site in OXG-RCBH, Asn488 interacts with the xylose side chain of the substrate, whereas the -1 site is occupied by Tyr457 in XEG. To confirm the contribution of this residue to substrate specificity, Tyr457 was substituted by Gly in XEG. The wild-type XEG cleaved the oligoxyloglucan at a specific site; the Y457G variant cleaved the same substrate, but at various sites. Together, the absence of a loop in the cleft and the presence of bulky Tyr457 determine the substrate specificity of XEG. PMID- 19682301 TI - A large complex mediated by Moc1, Moc2 and Cpc2 regulates sexual differentiation in fission yeast. AB - Sexual differentiation in Schizosaccharomyces pombe is triggered by nutrient starvation and is downregulated by cAMP. Screening programs have identified the moc1/sds23, moc2/ded1, moc3 and moc4/zfs1 genes as inducers of sexual differentiation, even in the presence of elevated levels of cAMP. To investigate possible interactions among Moc1, Moc2, Moc3 and Moc4 proteins, we first screened for individual Moc-interacting proteins using the yeast two-hybrid system and verified the interactions with other Moc proteins. Using this screening process, Cpc2 and Rpl32-2 were highlighted as factors involved in interactions with multiple Moc proteins. Cpc2 interacted with Moc1, Moc2 and Moc3, whereas the ribosomal protein Rpl32-2 interacted with all Moc proteins in the two-hybrid system. Physical interactions of Cpc2 with Moc1, Moc2 and Rpl32-2, and of Rpl32-2 with Moc2 were confirmed by coimmunoprecipitation. In addition, using Blue Native/PAGE, we revealed that each Moc protein exists as a large complex. Overexpression of Moc1, Moc2, Moc3, Moc4 and Rpl32-2 resulted in the efficient induction of a key transcription factor Ste11, suggesting that all proteins tested are positive regulators of Ste11. Considering that Moc2/Ded1 is a general translation factor and that Cpc2 associates with many ribosomal proteins, including Rpl32-2, it is possible that a large Moc-mediated complex, detected in this study, may act as a translational regulator involved in the control of sexual differentiation in S. pombe through the induction of Ste11. PMID- 19682302 TI - Weak toxin WTX from Naja kaouthia cobra venom interacts with both nicotinic and muscarinic acetylcholine receptors. AB - Iodinated [125I] weak toxin from Naja kaouthia (WTX) cobra venom was injected into mice, and organ-specific binding was monitored. Relatively high levels of [125I]WTX were detected in the adrenal glands. Rat adrenal membranes were therefore used for analysis of [125I]WTX-binding sites. Specific [125I]WTX binding was partially inhibited by both alpha-cobratoxin, a blocker of the alpha7 and muscle-type nicotinic acetylcholine receptors (nAChRs), and by atropine, an antagonist of the muscarinic acetylcholine receptor (mAChR). Binding to rat adrenal nAChR had a Kd of 2.0+/-0.8 microM and was inhibited by alpha-cobratoxin but not by a short-chain alpha-neurotoxin antagonist of the muscle-type nAChR, suggesting a specific interaction with the alpha7-type nAChR. WTX binding was reduced not only by atropine but also by other muscarinic agents (oxotremorine and muscarinic toxins from Dendroaspis angusticeps), indicating an interaction with mAChR. This interaction was further characterized using individual subtypes of human mAChRs expressed in Chinese hamster ovary cells. WTX concentrations up to 30 microM did not inhibit binding of [3H]acetylcholine to any subtype of mAChR by more than 50%. Depending on receptor subtype, WTX either increased or had no effect on the binding of the muscarinic antagonist [3H]N-methylscopolamine, which binds to the orthosteric site, a finding indicative of an allosteric interaction. Furthermore, WTX alone activated G-protein coupling with all mAChR subtypes and reduced the efficacy of acetylcholine in activating G-proteins with the M1, M4, and M5 subtypes. Our data demonstrate an orthosteric WTX interaction with nAChR and an allosteric interaction with mAChRs. PMID- 19682304 TI - The determinants of queen size in a socially polymorphic ant. AB - In social animals, body size can be shaped by multiple factors, such as direct genetic effects, maternal effects, or the social environment. In ants, the body size of queens correlates with the social structure of the colony: colonies headed by a single queen (monogyne) generally produce larger queens that are able to found colonies independently, whereas colonies headed by multiple queens (polygyne) tend to produce smaller queens that stay in their natal colony or disperse with workers. We performed a cross-fostering experiment to investigate the proximate causes of queen size variation in the socially polymorphic ant Formica selysi. As expected if genetic or maternal effects influence queen size, eggs originating from monogyne colonies developed into larger queens than eggs collected from polygyne colonies, be they raised by monogyne or polygyne workers. In contrast, eggs sampled in monogyne colonies were smaller than eggs sampled in polygyne colonies. Hence, eggs from monogyne colonies are smaller but develop into larger queens than eggs from polygyne colonies, independently of the social structure of the workers caring for the brood. These results demonstrate that a genetic polymorphism or maternal effect transmitted to the eggs influences queen size, which probably affects the social structure of new colonies. PMID- 19682303 TI - Energy barriers for HET-s prion forming domain amyloid formation. AB - The prion-forming domain comprising residues 218-289 of the fungal prion HET-s forms infectious amyloid fibrils at physiological pH. Because a high-resolution molecular model for the structure of these fibrils exists, it constitutes an attractive system with which to study the mechanism of amyloid assembly. Understanding aggregation under specific conditions requires a quantitative knowledge of the kinetics and thermodynamics of the self-assembly process. We report here the study of the temperature and agitation dependence of the HET s(218-289) fibril nucleation (kn) and elongation (ke) rate constants at physiological pH. Over our temperature and agitation range, kn and ke increased 30-fold and three-fold, respectively. Both processes followed the Arrhenius law, allowing calculation of the thermodynamic activation parameters associated with them. The data confirm the nucleation reaction as the rate-limiting step of amyloid fibril formation. The formation of the nucleus appears to depend mainly on enthalpic factors, whereas both enthalpic and entropic effects contribute similarly to the energy barrier to fibril elongation. A kinetic model is proposed in which nucleation depends on the presence of an initially collapsed, but poorly structured, HET-s(218-289) state and in which the fibril tip models the conformation of the incoming monomers without substantial disorganization of its structure during the elongation process. PMID- 19682305 TI - Pollen competition as an asymmetric reproductive barrier between two closely related Silene species. AB - Reproductive barriers are important determinants of gene flow between divergent populations or species. We studied pollen competition as a post-mating reproductive barrier between Silene dioica and S. latifolia. Gene flow between these species is extensive, but early-generation hybrids are rare. In an experiment with conspecific, heterospecific and 50 : 50 mixed pollinations in the two species, pollination treatments did not significantly affect seed set and seed weight. However, molecular determination of siring success after mixed pollinations showed that fewer than expected hybrids were produced in S. latifolia (18% hybrids) but not in S. dioica (51% hybrids). This constitutes an asymmetric post-mating reproductive barrier and likely contributes to the rarity of early-generation hybrids. Our study shows that pollen competition can be an effective barrier to hybridization between closely related species that likely acts in concert with other reproductive barriers. PMID- 19682306 TI - Evolution of sexual size dimorphism in grouse and allies (Aves: Phasianidae) in relation to mating competition, fecundity demands and resource division. AB - Sexual size dimorphism (SSD) is often assumed to be driven by three major selective processes: (1) sexual selection influencing male size and thus mating success, (2) fecundity selection acting on females and (3) inter-sexual resource division favouring different size in males and females to reduce competition for resources. Sexual selection should be particularly strong in species that exhibit lek polygyny, since male mating success is highly skewed in such species. We investigated whether these three selective processes are related to SSD evolution in grouse and allies (Phasianidae). Male-biased SSD increased with body size (Rensch's rule) and lekking species exhibited more male-biased SSD than nonlekking ones. Directional phylogenetic analyses indicated that lekking evolved before SSD, but conclusions were highly dependent on the body size traits and chosen model values. There was no relationship between SSD and male display agility, nor did resource division influence SSD. Although clutch mass increased with female body size it was not related to the degree of SSD. Taken together, the results are most consistent with the hypothesis that lekking behaviour led to the evolution of male-biased SSD in Phasianidae. PMID- 19682308 TI - Females benefit from multiple mating but not multiple mates in the burying beetle Nicrophorus vespilloides. AB - Male reproductive success generally increases with number of mates but this need not be true for females. If females are the limiting sex, as few as one mate can be optimal. Despite the theoretical differences driving multiple mating in the sexes, multiple mating is the norm rather than the exception. Empirical investigations are therefore required to determine why females mate with multiple males. Both nonadaptive (correlated responses to selection on males, given the mean mating rates have to be the same) and adaptive (direct or indirect fitness benefits) can drive the evolution of multiple mating in females. Females of the burying beetle Nicorphorus vespilloides often mate repeatedly with the same male, but this appears to be a correlated response to selection on males rather than reflecting direct benefits to females for multiple mating. However, an unexamined alternative to this nonadaptive explanation is that females benefit by mating with multiple different males and therefore are selected for general promiscuity. Here we examine if mating polyandrously provides fitness benefits by examining the effects of number of mates (1, 2 or 3), mating system (monogamous, polyandrous) and their interaction. The only significant influence was mating more than once. This did not depend on type of mating. We suggest that unlike most other species examined, in N. vespilloides mating with the same male repeatedly or with several different males reflects an indiscriminate willingness to mate as a result of correlated selection on males for high rates of mating. PMID- 19682307 TI - Phylogenetic and biological species diversity within the Neurospora tetrasperma complex. AB - The objective of this study was to explore the evolutionary history of the morphologically recognized filamentous ascomycete Neurospora tetrasperma, and to reveal the genetic and reproductive relationships among its individuals and populations. We applied both phylogenetic and biological species recognition to a collection of strains representing the geographic and genetic diversity of N. tetrasperma. First, we were able to confirm a monophyletic origin of N. tetrasperma. Furthermore, we found nine phylogenetic species within the morphospecies. When using the traditional broad biological species recognition all investigated strains of N. tetrasperma constituted a single biological species. In contrast, when using a quantitative measurement of the reproductive success, incorporating characters such as viability and fertility of offspring, we found a high congruence between the phylogenetic and biological species recognition. Taken together, phylogenetically and biologically defined groups of individuals exist in N. tetrasperma, and these should be taken into account in future studies of its life history traits. PMID- 19682309 TI - Heritability of resting metabolic rate in a wild population of blue tits. AB - We report the first study with the aim to estimate heritability in a wild population, a nest box breeding population of blue tits. We estimated heritability as well as genetic and phenotypic correlations of resting metabolic rate (RMR), body mass and tarsus length with an animal model based on data from a split cross-fostering experiment with brood size manipulations. RMR and body mass, but not tarsus length, showed significant levels of explained variation but for different underlying reasons. In body mass, the contribution to the explained variation is mainly because of a strong brood effect, while in RMR it is mainly because of a high heritability. The additive variance in RMR was significant and the heritability was estimated to 0.59. The estimates of heritability of body mass (0.08) and tarsus length (0.00) were both low and based on nonsignificant additive variances. Thus, given the low heritability (and additive variances) in body mass and tarsus length the potential for direct selection on RMR independent of the two traits is high in this population. However, the strong phenotypic correlation between RMR and mass (0.643 +/- 0.079) was partly accounted for by a potentially strong, although highly uncertain, genetic correlation (1.178 +/- 0.456) between the two traits. This indicates that the additive variance of body mass, although low, might still somewhat constrain the independent evolvability of RMR. PMID- 19682310 TI - Differential diagnosis between immune (idiopathic) thrombocytopenic purpura and portal vein thrombosis in children. PMID- 19682312 TI - Contralateral testicular relapse after prophylactic radiation in a patient with primary testicular diffuse large B-cell lymphoma. AB - Prophylactic radiation to the contralateral uninvolved testicle has become a standard practice in the treatment of primary testicular lymphoma. While it is generally felt to be very effective, its failure rate is unknown. We describe a patient with primary testicular diffuse large B-cell lymphoma, who developed recurrent disease in the contralateral testicle despite receiving prophylactic testicular radiation, central nervous system prophylaxis, and anthracycline-based chemo-immunotherapy. Review of the literature shows that the testicular failure rate after prophylactic radiation may be unexpectedly high at 10% or more. We put forward hypotheses on testicular relapse and discuss potential alternative preventive strategies. PMID- 19682313 TI - Connubial tinea gladiatorum due to Trichophyton mentagrophytes. AB - Trichophyton mentagrophytes is one of the most common dermatophytes causing cutaneous human fungal disease worldwide. Pubic and/or vulvar localisation of dermatophytosis has rarely been reported and may often be misdiagnosed as bacterial infections. We present a connubial case of severe inflammatory tinea in the genital region caused by T. mentagrophytes. The case illustrates the importance of getting material for cultivation before treatment and emphasises the difficulty of diagnosis and treatment of fungal infections in the genital region. PMID- 19682311 TI - FCRL2 mRNA expression is inversely associated with clinical progression in chronic lymphocytic leukemia. AB - Fc receptor-like 2 (FCRL2) is highly expressed on B-cell chronic lymphocytic leukemia (B-CLL) cells and could possibly influence disease pathogenesis. Therefore, we investigated FCRL2 mRNA expression in a large cohort with 152 CLL patients in order to assess its role in risk prediction in B-CLL. FCRL2 mRNA expression was found to be expressed at considerably higher levels in peripheral blood mononuclear cells (PBMC) of B-CLL patients compared to controls (range 1.35 to 210-fold upregulation; P < 0.0001) and cells of other hematological diseases. Patients with high FCRL2 expression (according to ROC-analysis) had a significantly longer treatment-free survival (TFS) and overall survival (OS) than patients with low FCRL2 expression (median TFS: 119 vs. 34 months, P < 0.0001; median OS: 321 months vs. not reached, P = 0.009). Univariate comparisons found that FCRL2 expression was weakly associated with IGHV mutation status (P = 0.05), CD38 status (P < 0.0001) and ZAP-70 status (P < 0.0001). Furthermore, we show that the combination of FCRL2 with ZAP70-, CD38- or IGHV-status could further significantly refine the prognostic information provided by either of the factors alone in TFS and OS. In multivariate analysis low FCRL2 expression was a significant independent prognostic factor (HR 2.4; P = 0.005). Here we demonstrate that the level of FCRL2 expression is correlated with prognosis in B CLL. PMID- 19682314 TI - Candida albicans or Candida dubliniensis? AB - Candida albicans is increasing as an opportunistic pathogen causing candidemia and candidiasis worldwide. In addition, other non-albicans Candida species are now also associated with pertinent infections. These include the closely related C. dubliniensis, which shares many phenotypic similarities with C. albicans. These similarities pose problems in the identification of isolates and have previously led to misidentification of these species. As a result, several identification techniques based on phenotypic and genotypic characteristics have been developed to differentiate between these Candida species. This review will focus on the similarities and differences between these two Candida species highlighting different identification methods and their advantages and disadvantages. PMID- 19682315 TI - Cutaneous zygomycosis caused by Saksenaea vasiformis following water-related wound in a 24-year-old immunocompetent woman. AB - We describe a case of cutaneous zygomycosis caused by Saksenaea vasiformis in an immunocompetent 24-year-old woman. Diagnosis was based on histological and microbiological examination. The patient made a complete recovery with surgical debridement and antifungal therapy (liposomal amphotericin and posaconazole). PMID- 19682316 TI - Mutation frequency of NS5A in patients vertically infected with HCV genotype 1 predicts sustained virological response to peginterferon alfa-2b and ribavirin combination therapy. AB - Viral genome analyses performed in adult HCV-patients yielded very inconsistent results and are not transferable to children who are often infected vertically during a state of high immune tolerance. We analysed the mutational frequency in the PKR-binding domain (PKR-BD) of NS5A and PePHD of E2 protein pre- and post treatment with peginterferon-alfa-2b and ribavirin in children chronically infected with HCV genotype 1. Amino acid sequences of NS5A (2 209-2 274) and E2 (618-681) were determined in serum samples using standard PCR procedures. Concerning the PKR-BD a significant higher number of mutations was observed in vertically compared to horizontally infected patients (2.14 vs 1.24, P-value = 0.03). This difference was exclusively based on the increased number of mutations in responders vs non-responders in vertically infected patients (2.95 vs 1.33; P value = 0.02). While all patients with at least four mutations (n = 3) did respond to therapy, no other predictive parameters could be identified. In the PePHD no differences could be observed between either of these groups. These findings support the idea that viral properties, mode and therewith time of infection in terms of immune tolerance are equally important factors for predicting SVR in children. However given the low number of cases further studies are required to confirm this hypothesis. PMID- 19682317 TI - Longitudinal evaluation of viral interactions in treated HIV-hepatitis B co infected patients with additional hepatitis C and D virus. AB - Virological interactions of hepatitis B (HBV), hepatitis C (HCV) and hepatitis D (HDV) viruses in HIV-infected patients have been poorly characterized especially under treatment influences. Undetection rates of hepatitis viruses were longitudinally analyzed in a 3-year cohort of 308 HIV-HBV co-infected patients and compared using Generalized Estimating Equation models adjusted for age, HIV RNA, CD4 cell-count and antiviral treatment. Chronic hepatitis co-infection in HIV-infected patients (age years, SD) was: 265 HBV (40.7, 8.2); 19 HBV-HCV (39.7, 4.1); 12 HBV-HDV (35.2, 9.9); 12 HBV-HCV-HDV (39.2, 5.2). At inclusion, treatment with lamivudine/tenofovir was not significantly different between co-infection groups. HBV suppression was significantly associated with HDV (aOR = 3.85, 95%CI 1.13-13.10, P = 0.03) and HCV tri-infection (aOR = 2.65, 95%CI 1.03-6.81, P = 0.04), but marginally associated with HIV-HBV-HCV-HDV (aOR = 2.32, 95%CI 0.94 5.74, P = 0.07). In quad-infection, lower HDV-undetectability (vs HIV-HBV-HDV, P = 0.2) and higher HCV-undetectability (vs HIV-HBV-HCV, P = 0.1) were demonstrated. The degree of HBV suppression varied between visits and co infection groups [range of aOR during follow-up (vs HIV-HBV co-infection): HIV HBV-HCV = 2.23-5.67, HIV-HBV-HDV = 1.53-15.17]. In treated co-infected patients, HDV expressed continuous suppression over HCV- and HBV-replications. Peaks and rebounds from undetectable hepatitis B, C and/or D viremia warrant closer follow up in this patient population. HDV-replication was uncontrolled even with antiviral treatment. PMID- 19682318 TI - Ki-67 expression in non-tumour epithelium adjacent to oral cancer as risk marker for multiple oral tumours. AB - OBJECTIVE: The aim of this study was to determine whether the differential assessment of epithelial proliferation is useful to diagnose premalignant fields and assess the risk of multiple tumours. MATERIAL AND METHODS: We analysed 83 oral carcinomas with associated non-tumour epithelium classified as distant or close according to its distance (> or <1 cm) from the invasion point, and as squamous hyperplasia, mild, moderate, severe dysplasia or carcinoma in situ. Twenty-five healthy oral mucosa samples were used as controls. An immunohistochemical technique was applied using Mib-1. Ki-67 in premalignant epithelium was assessed in basal layer, parabasal layer, medium and upper third. RESULTS: Parabasal expression was significantly higher or showed a tendency to be higher in close and distant epithelia with any histological grade than in the controls. Parabasal Ki-67 significantly differed between distant epithelia associated with multiple vs single tumours (P < 0.001) and between distant epithelia associated with multiple tumours vs controls (P < 0.001). This difference was not observed between distant epithelia associated with single tumours and controls (P = 0.175). The cut-off point that differentiated epithelia associated with multiple tumours was >50% of Ki-67 + parabasal cells in distant epithelia, which yielded 0.88 sensitivity and 0.79 specificity. CONCLUSIONS: The concept of a precancerous field may be linked to an increase in the proliferative activity of parabasal cells. PMID- 19682319 TI - Melatonin in diseases of the oral cavity. AB - BACKGROUND: Melatonin is the principal secretory product of the pineal gland. It has immunomodulatory and antioxidant activities, stimulates the proliferation of collagen and osseous tissue and acts as a protector against cellular degeneration associated with aging and toxin exposure. Arising out of its antioxidant actions, melatonin protects against inflammatory processes and cellular damage caused by the toxic derivates of oxygen. As a result of these actions, melatonin may be useful as a co-adjuvant in the treatment of certain conditions of the oral cavity. METHODS: An extensive review of the scientific literature was carried out using PubMed, Science Direct, ISI Web of Knowledge and the Cochrane base. RESULTS: Melatonin, which is released into the saliva, may have important implications for oral diseases. Melatonin may have beneficial effects in certain oral pathologies including periodontal diseases, herpes viral infections and Candida, local inflammatory rocesses, xerostomia, oral ulcers and oral cancer. CONCLUSIONS: Melatonin may play a role in protecting the oral cavity from tissue damage caused by oxidative stress. The experimental evidence suggests that melatonin may have utility in the treatment of several common diseases of the oral cavity. However, more specific studies are necessary to extend the therapeutic possibilities to other oral diseases. PMID- 19682320 TI - ELISA test for p63 antibodies in chronic ulcerative stomatitis. AB - OBJECTIVE: To develop a novel test for chronic ulcerative stomatitis (CUS), a chronic immunologically mediated condition that produces oral ulcerations. Current diagnostic methods require expensive and technically demanding in situ immunofluorescence (IF) studies. DESIGN: An Enzyme-Linked ImmunoSorbent Assay (ELISA) was prepared and tested with serum samples from patients with CUS and negative controls. MATERIALS AND METHODS: The N-terminal portion of the CUS autoantigen, DeltaNp63alpha, was produced as a purified recombinant protein and used to coat ELISA plates. Sera from 25 patients with CUS and 16 negative controls were analyzed for reactive antibodies. The optimal cut-offs for positive and negative samples were determined. MAIN OUTCOME MEASURES: The optimal cut-off of 0.236 resulted in a sensitivity and specificity of the ELISA of 0.80 and 0.75, respectively (exact 95% confidence intervals, P-value of <0.001). RESULTS: The ELISA developed in this study provides a novel and reliable diagnostic assessment to distinguish CUS from other oral ulcerative diseases. CONCLUSIONS: Immunoassay will allow the true incidence and prevalence of CUS to be determined in future studies. When combined with clinical correlations, the ELISA results will facilitate the evaluation of the prognostic utility of antibody titers and allow correlation with treatment responses in individual CUS cases. PMID- 19682321 TI - Intrathecal ziconotide for neuropathic pain: a review. AB - Neuropathic pain is a considerable burden that affects activities of daily living. The management of neuropathic pain can be challenging because of multiple etiologies and complex manifestations. Ziconotide is a nonopioid intrathecal (IT) analgesic option for patients with neuropathic pain refractory to conventional treatments. The objective of this article is to review the published literature on ziconotide for the treatment of neuropathic pain. Relevant publications were identified through searches of all years of 6 databases, which included PubMed, EMBASE, and CINAHL. Search terms used were ziconotide, SNX-111, MVIIA, Prialt, and neuropathic pain. Publications were included if ziconotide was intrathecally administered (either alone or in combination with other IT agents) to treat neuropathic pain of any etiology and if pain assessment was an outcome measure. Data extracted included study design, IT drug doses, pain outcome measures, and adverse events (AEs). Twenty-eight articles met the inclusion criteria: 5 were preclinical studies and 23 were clinical studies. In the preclinical studies, ziconotide demonstrated antiallodynic effects on neuropathic pain. Data from double-blind, placebo-controlled (DBPC) trials indicated that patients with neuropathic pain reported a mean percent improvement in pain score with ziconotide monotherapy that ranged from 15.7% to 31.6%. A low starting dose and slow titration of ziconotide resulted in an improved safety profile in the aforementioned trials. Common AEs associated with ziconotide include nausea and/or vomiting, dizziness, confusion, urinary retention, and somnolence. Evidence from DBPC trials, open-label studies, case series, and case studies suggests that ziconotide, as either monotherapy or in combination with other IT drugs, is a potential therapeutic option for patients with refractory neuropathic pain. Additional studies are needed to establish the long-term efficacy and safety of ziconotide for neuropathic pain. PMID- 19682322 TI - Porphyrin and nonporphyrin photosensitizers in oncology: preclinical and clinical advances in photodynamic therapy. AB - Photodynamic therapy (PDT) is now a well-recognized modality for the treatment of cancer. While PDT has developed progressively over the last century, great advances have been observed in the field in recent years. The concept of dual selectivity of PDT agents is now widely accepted due to the relative specificity and selectivity of PDT along with the absence of harmful side effects often encountered with chemotherapy or radiotherapy. Traditionally, porphyrin-based photosensitizers have dominated the PDT field but these first generation photosensitizers have several disadvantages, with poor light absorption and cutaneous photosensitivity being the predominant side effects. As a result, the requirement for new photosensitizers, including second generation porphyrins and porphyrin derivatives as well as third generation photosensitizers has arisen, with the aim of alleviating the problems encountered with first generation porphyrins and improving the efficacy of PDT. The investigation of nonporphyrin photosensitizers for the development of novel PDT agents has been considerably less extensive than porphyrin-based compounds; however, structural modification of nonporphyrin photosensitizers has allowed for manipulation of the photochemotherapeutic properties. The aim of this review is to provide an insight into PDT photosensitizers clinically approved for application in oncology, as well as those which show significant potential in ongoing preclinical studies. PMID- 19682323 TI - Assessment of UV biological spectral weighting functions for phenolic metabolites and growth responses in silver birch seedlings. AB - In research concerning stratospheric ozone depletion, action spectra are used as biological spectral weighting functions (BSWFs) for describing the effects of UV radiation on plant responses. Our aim was to evaluate the appropriateness of six frequently used BSWFs that differ in effectiveness with increasing wavelength. The evaluation of action spectra was based on calculating the effective UV radiation doses according to 1-2) two formulations of the generalized plant action spectrum, 3) a spectrum for ultraviolet induced erythema in human skin, 4) a spectrum for the accumulation of a flavonol in Mesembryanthemum crystallinum, 5) a spectrum for DNA damage in alfalfa seedlings and 6) the plant growth action spectrum. We monitored effects of UV radiation on the concentration of individual UV absorbing metabolites and chlorophyll concentrations in leaves and growth responses of silver birch (Betula pendula) seedlings. Experiments were conducted outdoors using plastic films attenuating different parts of the UV spectrum. Chlorophyll concentrations and growth were not affected by the UV treatments. The response to UV radiation varied between and within groups of phenolics. In general, the observed responses of phenolic groups and individual flavonoids were best predicted by action spectra extending into the UV-A region with moderate effectiveness. PMID- 19682324 TI - A helical membrane-binding domain targets the Toxoplasma ROP2 family to the parasitophorous vacuole. AB - During invasion, the obligate intracellular pathogen, Toxoplasma gondii, secretes into its host cell a variety of effector molecules, several of which have been implicated in strain-specific variation in disease. The largest family of these effectors, defined by the canonical member ROP2, quickly associates with the nascent parasitophorous vacuole membrane (PVM) after secretion. Here we demonstrate that the NH(2)-terminal domain of the ROP2 family contains a series of amphipathic helices that are necessary and sufficient for membrane association. While each of the amphipathic helices is individually competent to bind cellular membranes, together they act to bind the PVM preferentially, possibly through sensing its strong negative curvature. This previously uncharacterized helical domain is an evolutionarily robust and energetically efficient design for membrane association. PMID- 19682325 TI - Neuropathological correlates of lower limb corticobasal degeneration. PMID- 19682327 TI - Diffusive movement of processive kinesin-1 on microtubules. AB - The processive motor kinesin-1 moves unidirectionally toward the plus end of microtubules. This process can be visualized by total internal reflection fluorescence microscopy of kinesin bound to a carboxylated quantum dot (Qdot), which acts both as cargo and label. Surprisingly, when kinesin is bound to an anti-HIS Qdot, it shows diffusive movement on microtubules, which decreased in favor of processive runs with increasing salt concentration. This observation implies that kinesin movement on microtubules is governed by its conformation, as it is well established that kinesin undergoes a salt-dependent transition from a folded (inactive) to an extended (active) molecule. A truncated kinesin lacking the last 75 amino acids (kinesin-Delta C) showed both processive and diffusive movement on microtubules. The extent of each behavior depends on the relative amounts of ADP and ATP, with purely diffusive movement occurring in ADP alone. Taken together, these data imply that folded kinesin.ADP can exist in a state that diffuses along the microtubule lattice without expending energy. This mechanism may facilitate the ability of kinesin to pick up cargo, and/or allow the kinesin/cargo complex to stay bound after encountering obstacles. PMID- 19682328 TI - MHC II in dendritic cells is targeted to lysosomes or T cell-induced exosomes via distinct multivesicular body pathways. AB - Dendritic cells (DCs) express major histocompatibility complex class II (MHC II) to present peptide antigens to T cells. In immature DCs, which bear low cell surface levels of MHC II, peptide-loaded MHC II is ubiquitinated. Ubiquitination drives the endocytosis and sorting of MHC II to the luminal vesicles of multivesicular bodies (MVBs) for lysosomal degradation. Ubiquitination of MHC II is abrogated in activated DCs, resulting in an increased cell surface expression. We here provide evidence for an alternative MVB sorting mechanism for MHC II in antigen-loaded DCs, which is triggered by cognately interacting antigen-specific CD4+ T cells. At these conditions, DCs generate MVBs with MHC II and CD9 carrying luminal vesicles that are secreted as exosomes and transferred to the interacting T cells. Sorting of MHC II into exosomes was, in contrast to lysosomal targeting, independent of MHC II ubiquitination but rather correlated with its incorporation into CD9 containing detergent-resistant membranes. Together, these data indicate two distinct MVB pathways: one for lysosomal targeting and the other for exosome secretion. PMID- 19682326 TI - Does Alzheimer's disease begin in the brainstem? AB - Although substantial evidence indicates that the progression of pathological changes of the neuronal cytoskeleton is crucial in determining the severity of dementia in Alzheimer's disease (AD), the exact causes and evolution of these changes, the initial site at which they begin, and the neuronal susceptibility levels for their development are poorly understood. The current clinical criteria for diagnosis of AD are focused mostly on cognitive deficits produced by dysfunction of hippocampal and high-order neocortical areas, whereas noncognitive, behavioural and psychological symptoms of dementia such as disturbances in mood, emotion, appetite, and wake-sleep cycle, confusion, agitation and depression have been less considered. The early occurrence of these symptoms suggests brainstem involvement, and more specifically of the serotonergic nuclei. In spite of the fact that the Braak and Braak staging system and National Institutes of Aging - Reagan Institute (NIA-RI) criteria do not include their evaluation, several recent reports drew attention to the possibility of selective and early involvement of raphe nuclei, particularly the dorsal raphe nucleus (DRN), in the pathogenesis of AD. Based on these findings of differential susceptibility and anatomical connectivity, a novel pathogenetic scheme of AD progression was proposed. Although the precise mechanisms of neurofibrillary degeneration still await elucidation, we speculated that cumulative oxidative damage may be the main cause of DRN alterations, as the age is the main risk factor for sporadic AD. Within such a framework, beta-amyloid production is considered only as one of the factors (although a significant one in familial cases) that promotes molecular series of events underlying AD-related neuropathological changes. PMID- 19682329 TI - Transferrin-directed internalization and cycling of transferrin receptor 2. AB - Transferrin receptor 2 (TfR2) is a homologue of transferrin receptor 1 (TfR1) but has distinct functions from TfR1 in iron homeostasis. In keeping with its proposed role in iron sensing, previous studies showed that TfR2 has a short half life and that holo-Tf stabilizes TfR2 by redirecting it from a degradative pathway to a recycling pathway. In this study, we characterized how the endocytosis, recycling and degradation of TfR2 relates to its function and differs from TfR1. TfR2 endocytosis was adaptor protein-2 (AP-2) dependent. Flow cytometry analysis showed that TfR1 and TfR2 utilized the same endocytic pathway only in the presence of holo-Tf, indicating that holo-Tf alters the interaction of TfR2 with the endocytic machinery. Unlike TfR1, phosphofurin acidic cluster sorting protein 1 (PACS-1) binds to the cytoplasmic domain of TfR2 and data suggest that PACS-1 is involved in the TfR2 recycling. Depletion of TSG101 by siRNA or expression of a dominant negative Vps4 inhibited TfR2 degradation, indicating that TfR2 degradation occurs through a multivesicular body (MVB) pathway. TfR2 degradation is not mediated through ubiquitination on the single lysine (K31) in the cytoplasmic domain or on the amino terminal residue. No ubiquitination of TfR2 by HA-ubiquitin was detected, indicating a lack of direct TfR2 ubiquitination involvement in its degradation. PMID- 19682331 TI - Bacterial contamination of pediatric whole blood transfusions in a Kenyan hospital. AB - BACKGROUND: Hospitalized children in sub-Saharan Africa frequently receive whole blood transfusions for severe anemia. The risk from bacterial contamination of blood for transfusion in sub-Saharan Africa is not known. This study assessed the frequency of bacterial contamination of pediatric whole blood transfusions at a referral hospital in Kenya. STUDY DESIGN AND METHODS: This was an observational study. Over the course of 1 year, bacteriologic cultures were performed on 434 of the 799 blood packs issued to children by the blood bank of Coast Provincial General Hospital, Mombasa. Clinical outcome was not assessed. RESULTS: Forty-four bacterial contaminants were isolated from 38 blood packs-an overall contamination frequency of 8.8% (95% confidence interval, 6.1%-11.4%). Sixty-four percent of the bacteria isolated were Gram-negative. Many of the isolates are usually found in the environment and the most likely source of contamination was considered to be the hospital blood bank. CONCLUSION: Bacterial contamination of whole blood may be a significant but unrecognized hazard of blood transfusion for children in sub-Saharan Africa. Further work is needed to clarify the extent of the problem and its clinical consequences. Increased awareness and adherence to basic principles of asepsis in the hospital blood bank may be important immediate interventions. PMID- 19682332 TI - Efficacy of hepatitis B virus (HBV) DNA screening and characterization of acute and occult HBV infections among blood donors from Madrid, Spain. AB - BACKGROUND: Screening of blood units for hepatitis B virus (HBV) DNA identifies donations collected during the window period (WP) of the acute infection and may improve viral safety of the blood supply. It also leads to the detection of occult hepatitis B infection (OBI). STUDY DESIGN AND METHODS: From January 2005 to December 2006, a total of 383,267 blood units were screened for hepatitis B surface antigen (HBsAg) and HBV DNA in two transfusion centers in Madrid, using either individual-donation nucleic acid testing (ID-NAT) or minipool (MP-NAT) of eight donations (MP8). Samples positive for HBV DNA and negative for HBsAg were confirmed by a second molecular test, the viral DNA was quantified, and a genome fragment including the region encoding the major hydrophilic region (MHR) of HBsAg was sequenced. RESULTS: The overall yield of HBV DNA-positive, HBsAg negative units was 1 in 21,282 (18 cases), higher when using ID-NAT than MP8-NAT (1:9862 vs. 1:51,011; p < 0.01). Four donations (1/95,817) were collected during the infectious pre-HBsAg WP, one during an early recovery stage, and the remaining 13 (1/29,482) were OBIs, six of whom had no detectable antibody to HBsAg. Low-level Genotype D HBV DNA was detected in all OBI cases; the frequencies of this genotype and MHR amino acid substitutions were significantly higher than reported from unselected Spanish HBsAg carriers. Donors with OBI had normal aminotransferase levels and were significantly older than donors carrying HBsAg. CONCLUSIONS: Blood donors in the WP and with OBI are not uncommon in Madrid and are detected at a higher frequency with ID-NAT than MP-NAT. PMID- 19682333 TI - Instituting a thawed plasma procedure: it just makes sense and saves cents. AB - BACKGROUND: The objectives of this time-series study were to elucidate the impact of a thawed plasma standard operating procedure (TP SOP) on plasma wastage and on cost savings. STUDY DESIGN AND METHODS: This study compared plasma wastage for 1 year before versus 1 year after implementation of a TP SOP. RESULTS: The plasma wastage and discard declined 79.7 and 64.9%, respectively, with a cost savings of $15,654.79 during the 1 year after implementation of the TP SOP. The risk that a unit of plasma would be wasted decreased 86.2% from Year 1 to Year 2 and the risk that a unit of plasma would be discarded decreased 76.3% from Year 1 to Year 2. CONCLUSION: Our study showed the positive, sustained, impact of implementing a TP SOP. Twelve months after introducing the SOP our Blood Bank and Transfusion Medicine Services' plasma wastage and discard were dramatically reduced, saving thousands of dollars. Initiating a TP SOP just makes sense; it is easy to implement, conserves plasma, and saves cents. PMID- 19682334 TI - The characterization and classification of concurrent blood group antibodies. AB - BACKGROUND: Concurrent alloantibodies are defined as two or more blood group (BG) antibodies coexisting in a given patient. These antibodies are significant because they can present major problems in compatibility testing. The goals of this study were to determine the properties of concurrent BG antibodies. STUDY DESIGN AND METHODS: The transfusion records of 18,750 patients at a Veterans Affairs medical center were reviewed to identify alloimmunized individuals. The following data were collected on patients making concurrent antibodies: antibody specificities, time of first detection, whether the antibodies disappeared over time, and, if so, their time of disappearance. RESULTS: Multiple alloimmunization occurred in 21.7% (96/443) of alloimmunized patients, constituting 39.9% (230/577) of all antibodies. The rate at which an antibody was concurrent with another antibody varied by antigenic specificity (p < 0.05). Anti-C (21/28; 71.4%) and anti-c (19/27; 70.4%) were most likely to coexist with another antibody, while anti-P(1) (3/22; 13.6%) and anti-M (3/18; 16.7%) were least likely. The most common alloantibody pairs were anti-K/-E, anti-D/-C, and anti-E/ c. Paired antibodies were initially detected in the same antibody screen in most cases. The majority of antibody pairs were either both persistent or both evanescent. When they were evanescent, both antibodies usually became undetectable at the same time. The persistence over time of concurrent alloantibodies (70.4%) was not higher than that of antibodies occurring singly (67.5%; p = 0.63). CONCLUSIONS: Antibody concurrence varied by BG antigenic specificity. Paired BG antibodies tended to appear and disappear in a coordinated fashion. Concurrent alloimmunization had no effect on antibody persistence. PMID- 19682335 TI - A clinical scenario-based survey of transfusion decisions for intensive care patients with delayed weaning from mechanical ventilation. AB - BACKGROUND: Despite evidence supporting the use of restrictive hemoglobin (Hb) transfusion triggers in critically ill patients, translation of this evidence into practice remains inconsistent. It was hypothesized that clinicians believe that longer-term ventilated patients require a higher Hb, particularly when ischemic heart disease coexists. STUDY DESIGN AND METHODS: A scenario was developed describing an anemic patient recovering from multiple organ failure, but failing weaning trials after 6 days of mechanical ventilation. Clinicians were asked to state their Hb transfusion trigger and target Hb range assuming no history of ischemic heart disease (Scenario 1), known stable chronic ischemic heart disease (Scenario 2), or evidence of myocardial ischemia during weaning trials (Scenario 3). A prospective cross-sectional postal survey of clinicians practicing intensive care in the United Kingdom was undertaken. RESULTS: A total of 184 responses were obtained (52% response rate), which varied widely. Median (first, third quartile) transfusion trigger Hb levels were 8 (7, 8), 9 (8, 9.5), and 9.5 (9, 9.5) g/dL for Scenarios 1 to 3, respectively (p < 0.001 across and between each group). The target Hb was more than 9 g/dL for 47, 80, and 94% of respondents for Scenarios 1 to 3, respectively, and more than 10 g/dL for 14, 44, and 65% of respondents for Scenarios 1 to 3, respectively (p < 0.001 across the groups). CONCLUSIONS: In response to scenarios, clinicians in the United Kingdom believe that a more liberal transfusion practice is required for patients failing weaning trials after 6 days of mechanical ventilation than the current evidence base supports. PMID- 19682336 TI - Thrombin generation in trauma patients. AB - BACKGROUND: Trauma patients are at risk of developing an acute coagulopathy of trauma (ACT) related to tissue injury, shock, and hemodilution. ACT is incompletely understood, but is similar to disseminated intravascular coagulation (DIC) and is associated with poor outcome. STUDY DESIGN AND METHODS: Thrombin generation assays were used to evaluate plasma hemostasis in 42 trauma patients, 25 normal subjects, and 45 patients on warfarin and in laboratory-prepared factor reduced plasma. RESULTS: Prolonged prothrombin time (PT), more than 18 seconds, or an international normalized ratio of greater than 1.5 was present in 15 trauma patients indicating possible ACT. Native thrombin generation (no activator added, contact activation blocked) showed that Trauma with ACT patients had lag times 68% shorter and peak thrombin generation threefold higher than normal patients indicating the presence of circulating procoagulants capable of initiating coagulation systemically. Trauma patients had lower platelet counts and fibrinogen and Factor (F)II levels putting them at increased risk of bleeding. In laboratory-prepared isolated factor-reduced samples and in patients with vitamin K-dependent factor deficiency due to warfarin, thrombin generation decreased in direct proportion to FII levels. In contrast, in diluted plasma and in trauma patients with reduced factor levels, thrombin generation was increased and associated with slower inhibition of thrombin generation (prolonged termination time) and decreased antithrombin levels (43% of normal in Trauma with ACT). CONCLUSIONS: Thrombin generation studies indicate that Trauma with ACT patients show dysregulated hemostasis characterized by excessive non-wound-related thrombin generation due to a combination of circulating procoagulants capable of activating coagulation systemically and reduced inhibitor levels allowing systemic thrombin generation to continue once started. PMID- 19682338 TI - A novel rapid and effective donor arm disinfection method. AB - BACKGROUND: The aim of the study was to derive a donor arm disinfection technique that was rapid, but with a disinfection efficacy equivalent to a previous "best practice" technique. This method consisted of a two-stage procedure with an initial application of 70% isopropyl alcohol and then 2% tincture of iodine (IATI). The total time for the IATI method was 2 minutes in duration. A rapid technique (1 min in duration) was needed to obviate potential problems due to increased donor waiting time, had the IATI method been implemented at blood donation sessions. STUDY DESIGN AND METHODS: A direct swabbing and plating technique was used to enumerate bacteria present before and after disinfection. In total, seven methods were evaluated. RESULTS: The chlorhexidine/alcohol applicator (CAA) disinfection device containing 1.5 mL of 2% chlorhexidine gluconate and 70% isopropyl alcohol (99.91% reduction; confidence limits, 99.55%, 99.98%) was shown to have equivalent disinfection efficacy as the IATI method (99.89% reduction; confidence limits, 99.36%, 99.98%; p = 0.86). Procedural time for the 1.5-mL CAA method was 1 minute thereby avoiding potential problems of increased donor waiting time, inherent in the IATI 2-minute procedure at blood donation sessions. CONCLUSIONS: The 1.5-mL CAA disinfection method offers blood services a rapid and effective donor arm disinfection procedure. In 2006, the 1.5 mL CAA procedure was implemented throughout the entire English blood service for all donations. PMID- 19682339 TI - Relative safety of pooled whole blood-derived versus single-donor (apheresis) platelets in the United States: a systematic review of disparate risks. AB - BACKGROUND: Risks of transfusion-transmitted infections (TTIs), transfusion associated sepsis (TAS), and transfusion-related acute lung injury (TRALI) were compared between pooled whole blood-derived (PWBD) and single-donor platelets (PLTs) transfused in the United States. STUDY DESIGN AND METHODS: The literature was searched for estimates of the risk of TTIs and TAS and of the effect on bacterial contamination of PLTs of process improvements, bacterial culture, and surrogate methods to detect bacteria. Seven studies published between January 2005 and December 2008 and comparing bacterial contamination frequency between PWBD and single-donor PLTs after implementing bacterial culture testing of both components were subjected to meta-analysis. The three retrieved studies diagnosing TRALI based on the 2004 consensus definition in settings transfusing both PWBD and single-donor PLTs were not amenable to meta-analysis and were assessed qualitatively. RESULTS: Under a best-case scenario, if 100% (from the current 12.5%) of PLT doses were provided as PWBD PLTs, the number of additional transmissions of human immunodeficiency virus, hepatitis C virus, hepatitis B virus, bacteria, or a novel pathogen annually could be 1.2, 1.3, 9.0, 105.3, or 69.2 to 252.6, respectively. Compared with single-donor PLTs, US PLT pools of five concentrates have a 5.6-fold higher risk of bacterial contamination (summary odds ratio, 5.58; 95% confidence interval, 2.60-11.98; p < 0.05). The three studies that diagnosed TRALI based on the consensus definition did not demonstrate a difference in risk between PWBD and single-donor PLTs. CONCLUSIONS: TTIs and TAS determine the relative safety of PWBD versus single-donor PLTs. The available limited data do not support a higher risk of TRALI from single-donor (compared with PWBD) PLTs. PMID- 19682340 TI - A novel approach to pathogen reduction in platelet concentrates using short-wave ultraviolet light. AB - BACKGROUND: Transfusion of platelet concentrates (PCs) is the basic treatment for severe platelet disorders. PCs carry the risk of pathogen transmission, especially bacteria. Pathogen reduction (PR) by addition of photochemical reagents and irradiation with visible or ultraviolet (UV) light can significantly reduce this risk. We present a novel approach for PR in PCs employing UVC light alone. STUDY DESIGN AND METHODS: UVC PR was evaluated by bacteria and virus infectivity assays. PC quality was investigated by measuring pH, lactate, glucose, hypotonic shock response, platelet aggregation, CD62P expression, and annexin V binding as in vitro parameters. The impact of UVC PR on the platelet proteome was assessed by differential in-gel electrophoresis and compared with changes caused by UVB and gamma-irradiation, respectively. RESULTS: Vigorous agitation of loosely placed PCs generated thin fluid layers that allow penetration of UVC light for inactivation of the six bacteria and six of the seven virus species tested. HIV-1 was only moderately inactivated. UVC light at the dose used (0.4 J/cm(2)) had a minor impact on in vitro parameters and on storage stability of treated PCs. Proteome analysis revealed a common set of 92 (out of 793) protein spots being affected by all three types of irradiation. Specific alterations were most pronounced for gamma-irradiation (45 spots), followed by UVB (11 spots) and UVC (2 spots). CONCLUSION: UVC irradiation is a potential new method for pathogen reduction in PCs. The data obtained until now justify further development of this process. PMID- 19682337 TI - Understanding loss of donor white blood cell immunogenicity after pathogen reduction: mechanisms of action in ultraviolet illumination and riboflavin treatment. AB - BACKGROUND: Donor white blood cells (WBCs) present in transfusion products can lead to immune sequelae such as production of HLA antibodies or graft-versus-host disease in susceptible transfusion recipients. Eliminating the immunogenicity of blood products may prove to be of clinical benefit, particularly in patients requiring multiple transfusions in whom allosensitization is common. This study examines a method of pathogen reduction based on ultraviolet light illumination in the presence of riboflavin. In addition to pathogens, WBCs treated with this system are affected and fail to stimulate proliferation of allogeneic peripheral blood mononuclear cells (PBMNCs) in vitro. STUDY DESIGN AND METHODS: This study sought to determine the mechanisms regulating this loss of immunogenicity. Treated cells were examined for surface expression of a number of molecules involved in activation and adhesion, viability, cell-cell conjugation, and ability to stimulate immune responses in allogeneic PBMNCs. RESULTS: Compared with untreated controls, ultraviolet (UV)-irradiated antigen-presenting cells showed slightly reduced surface expression of HLA Class II and costimulatory molecules and had more significant reductions in surface expression of a number of adhesion molecules. Furthermore, treated cells had a severe defect in cell cell conjugation. The observed loss of immunogenicity was nearly complete, with UV-irradiated cells stimulating barely measurable interferon-gamma production and no detectable STAT-3, STAT-5, or CD3-epsilon phosphorylation in allospecific primed T cells. CONCLUSION: These results suggest that defective cell-cell adhesion prevents UV-irradiated cells from inducing T-cell activation. PMID- 19682341 TI - Coagulopathy screening and early plasma treatment for the prevention of intraventricular hemorrhage in preterm infants. AB - BACKGROUND: Despite the improvement in the assistance and treatment of preterm infants, intraventricular hemorrhage (IVH) remains a frequent complication in these patients. Our aim was to demonstrate the hypothesis that a coagulopathy screening and the early treatment with fresh-frozen plasma (FFP) of proven coagulopathy may contribute to decrease the occurrence of IVH in infants with gestational age of less than 29 weeks. STUDY DESIGN AND METHODS: This study compared two cohorts of infants who received FFP (10 mL/kg) after the evidence of pathologic coagulation tests performed within 2 hours after birth (screening group, n = 127) or after the development of bleedings and evidence of pathologic coagulation tests (no-screening group, n = 91). RESULTS: The screening strategy decreased the relative risk (RR) of developing IVH (RR = 0.65; 95% confidence interval, 0.44-0.98) compared to no-screening strategy, but the effect was limited to infants born at 23 to 26 weeks of gestation in whom screening strategy lowered IVH occurrence (34.5% vs. 61.1%, p = 0.008). CONCLUSIONS: A coagulopathy screening strategy decreases the risk of developing IVH in preterm infants but this effect is limited to infants born at 23 to 26 weeks of gestation. PMID- 19682342 TI - Large-dose intravenous ferric carboxymaltose injection for iron deficiency anemia in heavy uterine bleeding: a randomized, controlled trial. AB - BACKGROUND: The objective was to evaluate efficacy and safety of rapid, large dose intravenous (IV) administration of ferric carboxymaltose compared to oral iron in correcting iron deficiency anemia due to heavy uterine bleeding. STUDY DESIGN AND METHODS: In a randomized, controlled trial, 477 women with anemia, iron deficiency, and heavy uterine bleeding were assigned to receive either IV ferric carboxymaltose (or= 12 g/dL) of anemia (73% vs. 50%, p < 0.001). Patients treated with ferric carboxymaltose compared to those prescribed ferrous sulfate reported greater gains in vitality and physical function and experienced greater improvement in symptoms of fatigue (p < 0.05). There were no serious adverse drug events. CONCLUSIONS: In patients with iron deficiency anemia due to heavy uterine bleeding, rapid IV administration of large doses of a new iron agent, ferric carboxymaltose, is more effective than oral iron therapy in correcting anemia, replenishing iron stores, and improving quality of life. PMID- 19682343 TI - A new one-platform flow cytometric method for residual cell counting in platelet concentrates. AB - BACKGROUND: According to German regulations and guidelines, residual red blood cells (rRBCs) and residual white blood cells (rWBCs) must number fewer than 3 x 10(9) cells/unit and 1 x 10(6) cells/unit in platelet concentrates (PCs), respectively. Due to low levels of residual cells in final products, there is still a need for fast, reliable, and sensitive methods of automated detection of these cell types. STUDY DESIGN AND METHODS: In Part A, 21 PCs were spiked with predetermined numbers of red blood cells (RBCs) and white blood cells (WBCs). The linearity, precision, and accuracy of the BD Thrombo Count assay (BD Biosciences Europe) were tested and validated according to international guidelines. Finally in Part B, 100 PCs prepared from pooled buffy coats were tested by the BD Thrombo Count assay and compared with other methods, including Nageotte (rWBCs) and Neubauer (rRBCs) counting chambers and the flow cytometric BD LeucoCOUNT (Becton Dickinson) assay (rWBCs). RESULTS: The unspecific background of blank PC samples was fewer than 0.02 cells/microL for WBCs and fewer than 34 cells/microL for RBCs (mean, 21). Linear regression and precision analyses of spiked PC samples were determined for both WBCs (r(2) = 0.992; range, 0.6-6.0 WBCs/microL) and RBCs (r(2) = 0.999; 800-8000 RBCs/microL). No carryover of cells or drift in results was detected in the automated sample acquisition mode. Analysis according to statistical methods of Bland and Altman demonstrated a high correlation between BD Thrombo Count and the Neubauer manual counting chamber. CONCLUSION: This novel flow cytometric test is a quick and reliable single-tube assay that has been demonstrated as a potential alternative for the existing manual microscopic counting procedures that are both time-consuming and laborious. PMID- 19682344 TI - Liver donor's age and recipient's serum creatinine predict blood component use during liver transplantation. AB - BACKGROUND: Excessive use of blood components during liver transplantation should be avoided because it has been associated with poor outcomes and it may stress blood bank resources. STUDY DESIGN AND METHODS: To determine preoperative predictors of excessive transfusion requirements in patients undergoing liver transplantation, the clinical records of 126 consecutive adult patients undergoing primary liver transplantation were retrospectively reviewed. Outcome variables included number of red blood cells (RBCs), plasma, and plateletpheresis components intraoperatively transfused. Univariate analyses of the following predictor variables were performed: recipient age, sex, ethnicity, height/weight, Model for End Stage Liver Disease score, year of transplant, previous abdominal surgery, hepatoma, wait-list time, standard recipient laboratory values obtained immediately before transplantation, cold ischemia time, donor age, sex, and height/weight. Multivariate analysis using logistic regression was used to build a model that best predicted how many blood components should be available before transplant. RESULTS: Donor age of more than 50 years old (odds ratio [OR], 2.8 95% confidence interval [CI], 1.3-6.0), and recipient serum creatinine (SCr) level of more than 1.3 mg/dL (OR, 3.8 95% CI, 1.6-8.9) were the only variables found to be predictive of RBC use in multivariate analysis. This model accurately predicted the use of more than 10 units of RBCs 79% of cases. Having both adverse factors present resulted in using more than one box in 80% of cases as compared to 44% of cases where only one or no adverse factor was present (p = 0.002). Further analyses showed a direct correlation between the number of RBCs transfused and plasma (r = 0.93) and plateletpheresis components (r = 0.74) transfused. [Corrections added after online publication 22-Jul-2009: OR updated from 3.8 to 2.8; CI from 1.6-8.9 to 1.3-6.0; OR from 2.8-3.8.] CONCLUSION: Liver donor's age and recipient's SCr are important in preoperatively predicting blood use during liver transplantation. PMID- 19682345 TI - Infectivity of human immunodeficiency virus-1, hepatitis C virus, and hepatitis B virus and risk of transmission by transfusion. PMID- 19682347 TI - Bacterial screening of platelet concentrates: results of 2 years active surveillance of transfused positive cultured units released as negative to date. AB - BACKGROUND: The BacT/ALERT system for bacterial monitoring of platelet concentrates (PCs) was introduced in the Netherlands in 2001. Samples are cultured for 7 days, and as a result of the short shelf-life of PCs, they are usually released as 'negative to date'. Therefore, some of the PCs have already been transfused at the moment of a positive signal in continued cultures in the BacT/Alert. It is unclear, however, whether these PCs are associated with more transfusion reactions. METHODS: During a 2-year period clinical data were collected from all patients who received PCs released as 'negative to date' but with a positive bacterial culture after being transfused. RESULTS: Data of 158 patients who received PCs with confirmed positive bacterial culture tests were analysed. Two patients developed a transfusion reaction. In both PCs, Propionibacterium was cultured. The imputability as related to the transfusion was classified as unlikely in both patients. CONCLUSION: Two of 158 transfusions of PCs released as 'negative to date', but with a confirmed positive BacT/ALERT result, were initially associated with transfusion reactions. However, the imputability of both reactions was low. PMID- 19682346 TI - Red blood cell transfusion in the treatment and management of anaemia: the search for the elusive transfusion trigger. AB - Therapeutic red blood cell (RBC) transfusion is widely utilized in the management of anaemia. Critically ill intensive care unit (ICU) patients in particular, as well as medical and haematology-oncology patients, are among the largest groups of users of RBC products. While anaemia is common in these patients, its treatment and management, including appropriate thresholds for RBC transfusion, remain controversial. We review here the function of RBCs in oxygen transport and physiology, with a view to their role in supporting and maintaining systemic tissue oxygenation. Adaptive and physiological compensatory mechanisms in the setting of anaemia are discussed, along with the limits of compensation. Finally, data from clinical studies will be examined in search of evidence for, or against, a clinically relevant transfusion trigger. PMID- 19682348 TI - Blood donor screening with cobas s 201/cobas TaqScreen MPX under routine conditions at German Red Cross institutes. AB - BACKGROUND: In 1997 the German Red Cross (GRC) blood donor services introduced mini-pool nucleic acid testing (NAT) for human immunodeficiency virus (HIV)-1, hepatitis C virus (HCV) and hepatitis B virus (HBV) to increase blood safety. With the new cobas s 201/cobas TaqScreen MPX, a fully automated extraction method and a multiplex amplification system specifically adapted to the needs of blood donation services is available. METHODS: The cobas s 201 system was evaluated at the GRC BTS locations Hagen, Springe and Frankfurt. In phase A, the analytical sensitivity for the detection of HBV, HCV and HIV-1 was investigated and in phase B, at least 60,000 samples at each test site were screened in parallel with the MPX test on s 201 system and the existing routine mini-pool NAT system to compare the diagnostic specificity and the diagnostic sensitivity. RESULTS: Comparable analytical sensitivities in a range of 1.6-3.6 IU/ml, 4.9-10.9 IU/ml and 14.7 26.6 IU/ml for HBV, HCV HIV, respectively, for the MPX test on s 201 system (95% probability based on probit analysis) were determined at all test sites. The diagnostic sensitivity was 99.8% and the diagnostic specificity was 99.85%. CONCLUSIONS: The MPX test on s 201 system is a fully automated NAT system suitable for routine blood donor screening. The analytical sensitivity as well as the diagnostic sensitivity fulfilled all requirements of the Paul Ehrlich Institute for blood donor screening in mini-pools up to 96 donations per pool. A major benefit of the automated NAT system is the reduced personnel time and the extensive complete barcode-controlled process documentation. PMID- 19682349 TI - An international quality control programme for PRISM chemiluminescent immunoassays in blood service and blood product laboratories. AB - BACKGROUND: Laboratories screening for blood-borne virus infections in blood and blood products are required by international standards and guidelines to ensure that their testing processes remain within control. An effective means of ensuring this aim is through participation in a quality control programme. Analyses of results from a quality control (QC) programme conducted for the Abbott PRISM (PRISM) assays are reported. MATERIALS AND METHODS: Laboratories participating in the National Serology Reference Laboratory, Australia's PRISM QC programme were provided with aliquots of a multimarker QC sample which were tested regularly in each PRISM subchannel. Test results were submitted to a single database using an Internet-based QC monitoring system, EDCNet. The QC test results submitted between 15 October 2001 and 5 March 2006 for each PRISM instrument and each lot of PRISM reagent were analysed to determine the imprecision and bias in each test system. RESULTS: A total of 157,404 test results from approximately 47,000 test runs submitted into the EDCNet database were analysed. Six batches of the multimarker QC samples were tested in 454 PRISM reagent lots. The coefficient of variation of QC sample test results ranged from 9.17 to 15.83%, 8.29 to 9.44%, 10.50 to 15.38% and 7.05 to 10.32% when tested in the PRISM anti-hepatitis C virus, anti-human immunodeficiency virus, anti-human T cell lymphotrophic virus and hepatitis B surface antigen assays, respectively. Analysis of QC test results reported from testing in the anti-HTLV assay detected one lot of reagent (10572HN00) which was identified to be an outlier using Tukey's filter. DISCUSSION: Analysis of test results of an external QC sample can be used as a statistical process control through ongoing measurement of imprecision. When laboratories test the same QC sample in the same assay and submit test results to a single database, the results can be compared and a measure of bias can be calculated. The resulting QC programme can offer detection of unexpected variation in the testing processes and the source of variation investigated. PMID- 19682350 TI - Microbiology, time course and clinical characteristics of infection in critically ill patients receiving packed red blood cell transfusion. AB - BACKGROUND AND OBJECTIVES: Packed red blood cell transfusion has been associated with increased infection in a variety of critically ill patient populations. We evaluated the microbiology and time course of infection in transfused patients in the intensive care unit (ICU) as no data exist on these parameters. MATERIALS AND METHODS: We performed a retrospective review of data for all patients admitted to a 24-bed medical-surgical ICU at Cooper University Hospital from July 2003 to September 2006 and entered in the Project Impact database. RESULTS: A total of 2432 patients were admitted during the study period, of which 609 underwent transfusion. Transfused patients were more likely to develop a nosocomial infection (10.5% vs. 4.9%, P < 0.001). ICU and hospital length of stay were longer in the transfused group (P < 0.001 for both). Mortality was also greater (13.1% vs. 8.7%, P = 0.001). Transfused patients had a shorter time from hospital admission to first infection (P < 0.001) and ICU admission to first infection (P < 0.001). Multivariate analysis confirmed transfusion as an independent risk factor for infection, mortality, hospital and ICU length of stay. Methicillin resistant Staphylococcus aureus, vancomycin-resistant enterococcus and Acinetobacter occurred more often in transfused patients. Acinetobacter accounted for a disproportionate share of infections among transfused patients (P < 0.001). CONCLUSIONS: Transfused ICU patients have a higher incidence of nosocomial infection and worse outcomes. Transfused patients had a shorter onset of infection. Acinetobacter infection appears to be particularly common among these patients. Further investigation is merited to better elucidate the mechanism for these findings and their therapeutic and clinical implications. PMID- 19682352 TI - The greenhouse gas emissions performance of cellulosic ethanol supply chains in Europe. AB - BACKGROUND: Calculating the greenhouse gas savings that may be attributed to biofuels is problematic because production systems are inherently complex and methods used to quantify savings are subjective. Differing approaches and interpretations have fuelled a debate about the environmental merit of biofuels, and consequently about the level of policy support that can be justified. This paper estimates and compares emissions from plausible supply chains for lignocellulosic ethanol production, exemplified using data specific to the UK and Sweden. The common elements that give rise to the greatest greenhouse gas emissions are identified and the sensitivity of total emissions to variations in these elements is estimated. The implications of including consequential impacts including indirect land-use change, and the effects of selecting alternative allocation methods on the interpretation of results are discussed. RESULTS: We find that the most important factors affecting supply chain emissions are the emissions embodied in biomass production, the use of electricity in the conversion process and potentially consequential impacts: indirect land-use change and fertiliser replacement. The large quantity of electricity consumed during enzyme manufacture suggests that enzymatic conversion processes may give rise to greater greenhouse gas emissions than the dilute acid conversion process, even though the dilute acid process has a somewhat lower ethanol yield. CONCLUSION: The lignocellulosic ethanol supply chains considered here all lead to greenhouse gas savings relative to gasoline An important caveat to this is that if lignocellulosic ethanol production uses feedstocks that lead to indirect land use change, or other significant consequential impacts, the benefit may be greatly reduced.Co-locating ethanol, electricity generation and enzyme production in a single facility may improve performance, particularly if this allows the number of energy intensive steps in enzyme production to be reduced, or if other process synergies are available. If biofuels policy in the EU remains contingent on favourable environmental performance then the multi-scale nature of bioenergy supply chains presents a genuine challenge. Lignocellulosic ethanol holds promise for emission reductions, but maximising greenhouse gas savings will not only require efficient supply chain design but also a better understanding of the spatial and temporal factors which affect overall performance. PMID- 19682353 TI - Combined therapy with cyclophosphamide and DNA preparation inhibits the tumor growth in mice. AB - BACKGROUND: When cyclophosphamide and preparations of fragmented exogenous genomic double stranded DNA were administered in sequence, the regressive effect on the tumor was synergic: this combined treatment had a more pronounced effect than cyclophosphamide alone. Our further studies demonstrated that exogenous DNA stimulated the maturation and specific activities of dendritic cells. This suggests that cyclophosphamide, combined with DNA, leads to an immune response to the tumors that were grafted into the subjects post treatment. METHODS: Three month old CBA/Lac mice were used in the experiments. The mice were injected with cyclosphamide (200 mkg per 1 kg body weight) and genomic DNA (of human, mouse or salmon sperm origin). The DNA was administered intraperitoneally or subcutaneously. After 23 to 60 days, one million tumor cells were intramuscularly grafted into the mice. In the final experiment, the mice were pre-immunized by subcutaneous injections of 20 million repeatedly thawed and frozen tumor cells. Changes in tumor growth were determined by multiplying the three perpendicular diameters (measured by caliper). Students' t-tests were used to determine the difference between tumor growth and average survival rate between the mouse groups and the controls. RESULTS: An analysis of varying treatments with cyclophosphamide and exogenous DNA, followed by tumor grafting, provided evidence that this combined treatment had an immunizing effect. This inhibitory effect in mice was analyzed in an experiment with the classical immunization of a tumor homogenate. The strongest inhibitory action on a transplanted graft was created through the following steps: cyclophosphamide at 200 mg/kg of body weight administered as a pretreatment; 6 mg fragmented exogenous DNA administered over the course of 3 days; tumor homogenate grafted 10 days following the final DNA injection. CONCLUSION: Fragmented exogenous DNA injected with cyclophosphamide inhibits the growth of tumors that are grafted to mice after this combined treatment. PMID- 19682351 TI - Two mycoheterotrophic orchids from Thailand tropical dipterocarpacean forests associate with a broad diversity of ectomycorrhizal fungi. AB - BACKGROUND: Mycoheterotrophic plants are considered to associate very specifically with fungi. Mycoheterotrophic orchids are mostly associated with ectomycorrhizal fungi in temperate regions, or with saprobes or parasites in tropical regions. Although most mycoheterotrophic orchids occur in the tropics, few studies have been devoted to them, and the main conclusions about their specificity have hitherto been drawn from their association with ectomycorrhizal fungi in temperate regions. RESULTS: We investigated three Asiatic Neottieae species from ectomycorrhizal forests in Thailand. We found that all were associated with ectomycorrhizal fungi, such as Thelephoraceae, Russulaceae and Sebacinales. Based on 13C enrichment of their biomass, they probably received their organic carbon from these fungi, as do mycoheterotrophic Neottieae from temperate regions. Moreover, 13C enrichment suggested that some nearby green orchids received part of their carbon from fungi too. Nevertheless, two of the three orchids presented a unique feature for mycoheterotrophic plants: they were not specifically associated with a narrow clade of fungi. Some orchid individuals were even associated with up to nine different fungi. CONCLUSION: Our results demonstrate that some green and mycoheterotrophic orchids in tropical regions can receive carbon from ectomycorrhizal fungi, and thus from trees. Our results reveal the absence of specificity in two mycoheterotrophic orchid-fungus associations in tropical regions, in contrast to most previous studies of mycoheterotrophic plants, which have been mainly focused on temperate orchids. PMID- 19682354 TI - Recruitment rates and reasons for community physicians' non-participation in an interdisciplinary intervention study on leg ulceration. AB - BACKGROUND: This article describes the challenges a research team experienced recruiting physicians within a randomised controlled trial about leg ulcer care that seeks to foster the cooperation between the medical and nursing professions. Community-based physicians in North Rhine-Westphalia, Germany, were recruited for an interdisciplinary intervention designed to enhance leg ulcer patients' self care agency. The aim of this article is to investigate the success of different recruitment strategies employed and reasons for physicians' non-participation. METHODS: The first recruitment phase stressed the recruitment of GPs, the second the recruitment of specialists. Throughout the recruitment process data were collected through phone conversations with GP practices who indicated reasons for non-participation. RESULTS: Despite great efforts to recruit physicians, the recruitment rate reached only 26 out of 1549 contacted practices (1.7%) and 12 out of 273 (4.4%) practices during the first and second recruitment phase respectively. The overall recruitment rate over the 16-month recruitment period was 2%. With a target recruitment rate of n = 300, only 45 patients were enrolled in the study, not meeting study projections. Various reasons for community physicians' non-participation are presented as stated spontaneously during phone conversations that might explain low recruitment rates. The recruitment strategy utilised is discussed against the background of factors associated with high participation rates from the international literature. CONCLUSION: Time, money, and effort needed during the planning and recruitment phase of a study must not be underestimated to avoid higher than usual rates of refusal and lack of initial contact. Pilot studies prior to a study start-up may provide some evidence on whether the target recruitment rate is feasible. TRIAL REGISTRATION: Current Controlled Trials ISRCTN42122226. PMID- 19682356 TI - Breast, cervical, and colorectal cancer screening rates amongst female Cambodian, Somali, and Vietnamese immigrants in the USA. AB - INTRODUCTION: Minority women, particularly immigrants, have lower cancer screening rates than Caucasian women, but little else is known about cancer screening among immigrant women. Our objective was to assess breast, cervical, and colorectal cancer screening rates among immigrant women from Cambodia, Somalia, and Vietnam and explore screening barriers. METHODS: We measured screening rates by systematic chart review (N = 100) and qualitatively explored screening barriers via face-to-face questionnaire (N = 15) of women aged 50-75 from Cambodia, Somalia, and Vietnam attending a general medicine clinic (Portland, Maine, USA). RESULTS: Chart Review - Somali women were at higher risk of being unscreened for breast, cervical, and colorectal cancer compared with Cambodian and Vietnamese women. A longer period of US residency was associated with being screened for colorectal cancer. We observed a 7% (OR 1.07, 95% CI 1.01 1.13, p = 0.01) increase in the odds that a woman would undergo a fecal occult blood test for each additional year in the US, and a 39% increase in the odds of a woman being screened by colonoscopy or flexible sigmoidoscopy for every five years of additional US residence (OR 1.39, 95% CI 1.21-1.61, p = 0.02). We did not observe statistically significant relationships between odds of being screened by mammography, clinical breast exam or papanicolaou test according to years in the US. Questionnaire - We identified several barriers to breast, cervical, and colorectal cancer screening, including discomfort with exams conducted by male physicians. DISCUSSION: Somali women were less likely to be screened for breast, cervical, and colorectal cancer than Cambodian and Vietnamese women in this population, and uptake of colorectal cancer screening is associated with years of residency in this country. Future efforts to improve equity in cancer screening among immigrants may require both provider and community education. PMID- 19682355 TI - Influence of gender, sexual orientation, and need on treatment utilization for substance use and mental disorders: findings from the California Quality of Life Survey. AB - BACKGROUND: Prior research has shown a higher prevalence of substance use and mental disorders among sexual minorities, however, the influence of sexual orientation on treatment seeking has not been widely studied. We use a model of help-seeking for vulnerable populations to investigate factors related to treatment for alcohol or drug use disorders and mental health disorders, focusing on the contributions of gender, sexual orientation, and need. METHODS: Survey data were obtained from a population-based probability sample of California residents that oversampled for sexual minorities. Logistic regression was used to model the enabling, predisposing, and need-related factors associated with past year mental health or substance abuse treatment utilization among adults aged 18 64 (N = 2,074). RESULTS: Compared with individuals without a diagnosed disorder, those with any disorder were more likely to receive treatment. After controlling for both presence of disorder and other factors, lesbians and bisexual women were most likely to receive treatment and heterosexual men were the least likely. Moreover, a considerable proportion of sexual orientation minorities without any diagnosable disorder, particularly lesbians and bisexual women, also reported receiving treatment. CONCLUSION: The study highlights the need to better understand the factors beyond meeting diagnostic criteria that underlie treatment utilization among sexual minorities. Future research should also aim to ascertain the effects of treatment provided to sexual minorities with and without diagnosable disorders, including the possibility that the provision of such treatment may reduce the likelihood of their progression to greater severity of distress, disorders, or impairments in functioning. PMID- 19682357 TI - Rapid detection of ERG11 gene mutations in clinical Candida albicans isolates with reduced susceptibility to fluconazole by rolling circle amplification and DNA sequencing. AB - BACKGROUND: Amino acid substitutions in the target enzyme Erg11p of azole antifungals contribute to clinically-relevant azole resistance in Candida albicans. A simple molecular method for rapid detection of ERG11 gene mutations would be an advantage as a screening tool to identify potentially-resistant strains and to track their movement. To complement DNA sequencing, we developed a padlock probe and rolling circle amplification (RCA)-based method to detect a series of mutations in the C. albicans ERG11 gene using "reference" azole resistant isolates with known mutations. The method was then used to estimate the frequency of ERG11 mutations and their type in 25 Australian clinical C. albicans isolates with reduced susceptibility to fluconazole and in 23 fluconazole susceptible isolates. RCA results were compared DNA sequencing. RESULTS: The RCA assay correctly identified all ERG11 mutations in eight "reference" C. albicans isolates. When applied to 48 test strains, the RCA method showed 100% agreement with DNA sequencing where an ERG11 mutation-specific probe was used. Of 20 different missense mutations detected by sequencing in 24 of 25 (96%) isolates with reduced fluconazole susceptibility, 16 were detected by RCA. Five missense mutations were detected by both methods in 18 of 23 (78%) fluconazole-susceptible strains. DNA sequencing revealed that mutations in non-susceptible isolates were all due to homozygous nucleotide changes. With the exception of the mutations leading to amino acid substitution E266D, those in fluconazole-susceptible strains were heterozygous. Amino acid substitutions common to both sets of isolates were D116E, E266D, K128T, V437I and V488I. Substitutions unique to isolates with reduced fluconazole susceptibility were G464 S (n = 4 isolates), G448E (n = 3), G307S (n = 3), K143R (n = 3) and Y123H, S405F and R467K (each n = 1). DNA sequencing revealed a novel substitution, G450V, in one isolate. CONCLUSION: The sensitive RCA assay described here is a simple, robust and rapid (2 h) method for the detection of ERG11 polymorphisms. It showed excellent concordance with ERG11 sequencing and is a potentially valuable tool to track the emergence and spread of azole-resistant C. albicans and to study the epidemiology of ERG11 mutations. The RCA method is applicable to the study of azole resistance in other fungi. PMID- 19682358 TI - Development and validation of a simple questionnaire for the identification of hereditary breast cancer in primary care. AB - BACKGROUND: Breast cancer is a significant public health problem worldwide and the development of tools to identify individuals at-risk for hereditary breast cancer syndromes, where specific interventions can be proposed to reduce risk, has become increasingly relevant. A previous study in Southern Brazil has shown that a family history suggestive of these syndromes may be prevalent at the primary care level. Development of a simple and sensitive instrument, easily applicable in primary care units, would be particularly helpful in underserved communities in which identification and referral of high-risk individuals is difficult. METHODS: A simple 7-question instrument about family history of breast, ovarian and colorectal cancer, FHS-7, was developed to screen for individuals with an increased risk for hereditary breast cancer syndromes. FHS-7 was applied to 9218 women during routine visits to primary care units in Southern Brazil. Two consecutive samples of 885 women and 910 women who answered positively to at least one question and negatively to all questions were included, respectively. The sensitivity, specificity and positive and negative predictive values were determined. RESULTS: Of the 885 women reporting a positive family history, 211 (23.8%; CI95%: 21.5-26.2) had a pedigree suggestive of a hereditary breast and/or breast and colorectal cancer syndrome. Using as cut point one positive answer, the sensitivity and specificity of the instrument were 87.6% and 56.4%, respectively. Concordance between answers in two different applications was given by a intra-class correlation (ICC) of 0.84 for at least one positive answer. Temporal stability of the instrument was adequate (ICC = 0.65). CONCLUSION: A simple instrument for the identification of the most common hereditary breast cancer syndrome phenotypes, showing good specificity and temporal stability was developed and could be used as a screening tool in primary care to refer at-risk individuals for genetic evaluations. PMID- 19682359 TI - Rapid reduction versus abrupt quitting for smokers who want to stop soon: a randomised controlled non-inferiority trial. AB - BACKGROUND: The standard way to stop smoking is to stop abruptly on a quit day with no prior reduction in consumption of cigarettes. Many smokers feel that reduction is natural and if reduction programmes were offered, many more might take up treatment. Few trials of reduction versus abrupt cessation have been completed. Most are small, do not use pharmacotherapy, and do not meet the standards necessary to obtain a marketing authorization for a pharmacotherapy. DESIGN/METHODS: We will conduct a non-inferiority randomised trial of rapid reduction versus standard abrupt cessation among smokers who want to stop smoking. In the reduction arm, participants will be advised to reduce smoking consumption by half in the first week and to 25% of baseline in the second, leading up to a quit day at which participants will stop smoking completely. This will be assisted by nicotine patches and an acute form of nicotine replacement therapy. In the abrupt arm participants will use nicotine patches only, whilst smoking as normal, for two weeks prior to a quit day, at which they will also stop smoking completely. Smokers in either arm will have standard withdrawal orientated behavioural support programme with a combination of nicotine patches and acute nicotine replacement therapy post-cessation. OUTCOMES/FOLLOW-UP: The primary outcome of interest will be prolonged abstinence from smoking, with secondary trial outcomes of point prevalence, urges to smoke and withdrawal symptoms. Follow up will take place at 4 weeks, 8 weeks and 6 months post-quit day. TRIAL REGISTRATION: Current Controlled Trials ISRCTN22526020. PMID- 19682361 TI - Endoscopic T-tube placement in the management of lye-induced esophageal perforation: Case report of a safe treatment strategy. AB - Esophageal perforation is associated with a significant risk of morbidity and mortality. We report herein a case of lye-induced esophageal perforation managed successfully by employing endoscopic T-tube placement with a successful outcome. PMID- 19682360 TI - Efficacy of prescribed injectable diacetylmorphine in the Andalusian trial: Bayesian analysis of responders and non-responders according to a multi domain outcome index. AB - BACKGROUND: The objective of this research was to evaluate data from a randomized clinical trial that tested injectable diacetylmorphine (DAM) and oral methadone (MMT) for substitution treatment, using a multi-domain dichotomous index, with a Bayesian approach. METHODS: Sixty two long-term, socially-excluded heroin injectors, not benefiting from available treatments were randomized to receive either DAM or MMT for 9 months in Granada, Spain. Completers were 44 and data at the end of the study period was obtained for 50. Participants were determined to be responders or non responders using a multi-domain outcome index accounting for their physical and mental health and psychosocial integration, used in a previous trial. Data was analyzed with Bayesian methods, using information from a similar study conducted in The Netherlands to select a priori distributions. On adding the data from the present study to update the a priori information, the distribution of the difference in response rates were obtained and used to build credibility intervals and relevant probability computations. RESULTS: In the experimental group (n = 27), the rate of responders to treatment was 70.4% (95% CI 53.287.6), and in the control group (n = 23), it was 34.8% (95% CI 15.354.3). The probability of success in the experimental group using the a posteriori distributions was higher after a proper sensitivity analysis. Almost the whole distribution of the rates difference (the one for diacetylmorphine minus methadone) was located to the right of the zero, indicating the superiority of the experimental treatment. CONCLUSION: The present analysis suggests a clinical superiority of injectable diacetylmorphine compared to oral methadone in the treatment of severely affected heroin injectors not benefiting sufficiently from the available treatments. TRIAL REGISTRATION: Current Controlled Trials ISRCTN52023186. PMID- 19682362 TI - Quality of paediatric blood transfusions in two district hospitals in Tanzania: a cross-sectional hospital based study. AB - BACKGROUND: Blood transfusion (BT) can be lifesaving for children; however, monitoring the quality of BT is important. The current study describes the quality of paediatric BT delivered in two district hospitals in north-east Tanzania in order to identify areas for quality assurance and improvement in the administration of BT. METHODS: All 166 children admitted in the paediatric wards and receiving BT through April to June 2007 were prospectively observed. Medical records, request forms and registers in the laboratories were reviewed to identify blood source, blood screening and indications for BT. BT was observation before, during and after transfusion process. RESULTS: Malaria related anaemia accounted for 98% of the BTs. Ninety-two percent of the children were assessed for paleness. Clinical signs such as difficult breathing and symptoms of cardiac failure were only assessed in 67% and 15% of the children respectively, prior to the BT decision. Pre-transfusion haemoglobin and body temperature were recorded in 2/3 of the patients, but respiratory rate and pulse rate were not routinely recorded. In 40% of BTs, the transfusion time exceeded the recommended 4 hours. The zonal blood bank (ZBB) and local donors accounted for 10% and 90% of the blood, respectively. ABO and RhD typing and screening for HIV and syphilis were undertaken in all transfused blood. Evidence for hepatitis B or C infection was not checked except in the ZBB. CONCLUSION: Criteria for BT are not always fulfilled; time to initiate and complete the transfusion is often unacceptable long and monitoring of vital signs during BT is poor. Blood from the ZBB was often not available and BT often depended on local donors which implied lack of screening for hepatitis B and C. It is recommended that an external supervision system be established to monitor and evaluate the quality of BT performance in the laboratories as well as in wards. PMID- 19682363 TI - Integration of irradiation with cytoplasmic incompatibility to facilitate a lymphatic filariasis vector elimination approach. AB - BACKGROUND: Mass drug administration (MDA) is the emphasis of an ongoing global lymphatic filariasis (LF) elimination program by the World Health Organization, in which the entire 'at risk' human population is treated annually with anti filarial drugs. However, there is evidence that the MDA strategy may not be equally appropriate in all areas of LF transmission, leading to calls for the augmentation of MDA with anti-vector interventions. One potential augmentative intervention is the elimination of vectors via repeated inundative releases of male mosquitoes made cytoplasmically incompatible via an infection with Wolbachia bacteria. However, with a reduction in the vector population size, there is the risk that an accidental female release would permit the establishment of the incompatible Wolbachia infection type, resulting in population replacement instead of population elimination. To avoid the release of fertile females, we propose the exposure of release individuals to low doses of radiation to sterilize any accidentally released females, reducing the risk of population replacement. RESULTS: Aedes polynesiensis pupae of differing ages were irradiated to determine a radiation dose that results in sterility but that does not affect the survival and competitiveness of males. Laboratory assays demonstrate that males irradiated at a female sterilizing dosage of 40 Gy are equally competitive with un-irradiated males. No effect of irradiation on the ability of Wolbachia to affect egg hatch was observed. CONCLUSION: An irradiation dose of 40 Gy is sufficient to cause female sterility, but has no observed negative effect on male fitness. The results support further development of this approach as a preventative measure against accidental population replacement. PMID- 19682365 TI - An XML transfer schema for exchange of genomic and genetic mapping data: implementation as a web service in a Taverna workflow. AB - BACKGROUND: Genomic analysis, particularly for less well-characterized organisms, is greatly assisted by performing comparative analyses between different types of genome maps and across species boundaries. Various providers publish a plethora of on-line resources collating genome mapping data from a multitude of species. Datasources range in scale and scope from small bespoke resources for particular organisms, through larger web-resources containing data from multiple species, to large-scale bioinformatics resources providing access to data derived from genome projects for model and non-model organisms. The heterogeneity of information held in these resources reflects both the technologies used to generate the data and the target users of each resource. Currently there is no common information exchange standard or protocol to enable access and integration of these disparate resources. Consequently data integration and comparison must be performed in an ad hoc manner. RESULTS: We have developed a simple generic XML schema (GenomicMappingData.xsd - GMD) to allow export and exchange of mapping data in a common lightweight XML document format. This schema represents the various types of data objects commonly described across mapping datasources and provides a mechanism for recording relationships between data objects. The schema is sufficiently generic to allow representation of any map type (for example genetic linkage maps, radiation hybrid maps, sequence maps and physical maps). It also provides mechanisms for recording data provenance and for cross referencing external datasources (including for example ENSEMBL, PubMed and Genbank.). The schema is extensible via the inclusion of additional datatypes, which can be achieved by importing further schemas, e.g. a schema defining relationship types. We have built demonstration web services that export data from our ArkDB database according to the GMD schema, facilitating the integration of data retrieval into Taverna workflows. CONCLUSION: The data exchange standard we present here provides a useful generic format for transfer and integration of genomic and genetic mapping data. The extensibility of our schema allows for inclusion of additional data and provides a mechanism for typing mapping objects via third party standards. Web services retrieving GMD-compliant mapping data demonstrate that use of this exchange standard provides a practical mechanism for achieving data integration, by facilitating syntactically and semantically-controlled access to the data. PMID- 19682364 TI - SNAD: Sequence Name Annotation-based Designer. AB - BACKGROUND: A growing diversity of biological data is tagged with unique identifiers (UIDs) associated with polynucleotides and proteins to ensure efficient computer-mediated data storage, maintenance, and processing. These identifiers, which are not informative for most people, are often substituted by biologically meaningful names in various presentations to facilitate utilization and dissemination of sequence-based knowledge. This substitution is commonly done manually that may be a tedious exercise prone to mistakes and omissions. RESULTS: Here we introduce SNAD (Sequence Name Annotation-based Designer) that mediates automatic conversion of sequence UIDs (associated with multiple alignment or phylogenetic tree, or supplied as plain text list) into biologically meaningful names and acronyms. This conversion is directed by precompiled or user-defined templates that exploit wealth of annotation available in cognate entries of external databases. Using examples, we demonstrate how this tool can be used to generate names for practical purposes, particularly in virology. CONCLUSION: A tool for controllable annotation-based conversion of sequence UIDs into biologically meaningful names and acronyms has been developed and placed into service, fostering links between quality of sequence annotation, and efficiency of communication and knowledge dissemination among researchers. PMID- 19682367 TI - Improved base calling for the Illumina Genome Analyzer using machine learning strategies. AB - The Illumina Genome Analyzer generates millions of short sequencing reads. We present Ibis (Improved base identification system), an accurate, fast and easy-to use base caller that significantly reduces the error rate and increases the output of usable reads. Ibis is faster and more robust with respect to chemistry and technology than other publicly available packages. Ibis is freely available under the GPL from http://bioinf.eva.mpg.de/Ibis/. PMID- 19682366 TI - Identification and expression of differentially expressed genes in the hard clam, Mercenaria mercenaria, in response to quahog parasite unknown (QPX). AB - BACKGROUND: The hard clam, Mercenaria mercenaria, has been affected by severe mortality episodes associated with the protistan parasite QPX (Quahog Parasite Unknown) for several years. Despite the commercial importance of hard clams in the United States, molecular bases of defense mechanisms in M. mercenaria, especially during QPX infection, remain unknown. RESULTS: Our study used suppression subtractive hybridization (SSH), as well as the construction of cDNA libraries from hemocytes to identify genes related to the defense of the hard clam against its parasite. Hard clams were experimentally infected with QPX and SSH was performed on mRNA samples extracted from mantle and gill tissues at different times post-challenge. A total of 298 clones from SSH libraries and 1352 clones from cDNA libraries were sequenced. Among these sequences, homologies with genes involved in different physiological processes related to signal transduction, stress response, immunity and protein synthesis were identified. Quantitative PCR revealed significant changes in the expression of several of these genes in response to QPX challenge and demonstrated significant correlations in terms of levels of gene expression between intermediates of signalling pathways and humoral defense factors, such as big defensin and lysozyme. CONCLUSION: Results of this study allowed the detection of modifications caused by QPX at the transcriptional level providing insight into clam immune response to the infection. These investigations permitted the identification of candidate genes and pathways for further analyses of biological bases of clam resistance to QPX allowing for a better understanding of bivalve immunity in general. PMID- 19682369 TI - Streptococcus pneumoniae serotype 3 among Costa Rican children with otitis media: clinical, epidemiological characteristics and antimicrobial resistance patterns. AB - BACKGROUND: After the introduction of the seven valent-pneumococcal conjugated vaccine into our National Immunization Program, it is important to establish and track local serotype distribution in order to evaluate its impact specially because serotype replacement phenomena has been described.To describe the clinical, epidemiological and antimicrobial resistance patterns of Costa Rican children with otitis media caused by Streptococcus pneumoniae serotype 3. METHODS: Middle ear fluid samples were obtained from Costa Rican children with otitis media who participated in various antimicrobial clinical trials between 1992 and 2007. Streptococcus pneumoniae was identified according to laboratory standard procedures. Strains were serotyped and antimicrobial susceptibility to penicillin, amoxicillin, cefuroxime, ceftriaxone, azithromycin and levofloxacin was determined by E-test. RESULTS: Throughout 1992-2007 a total of 1919 tympanocentesis were performed in children with otitis media (median age: 19 months) and yielded a total of 1208 middle ear isolates. The most common pathogens were: Streptococcus pneumoniae, 511 isolates (49%); Non-Typable Haemophilus influenzae, 386 isolates (37%); Moraxella catarrahalis, 100 isolates (9.5%); and Streptococcus pyogenes, 54 isolates (5%). Streptococcus pneumoniae serotyping was performed in 346/511 isolates (68%) recovered during years 1999 2006. The most common serotypes were 19F (101/30.0%), 14 (46/13.7%), 3 (34/10.1%), 6B (30/8.9%) and 23F (23/6.8%). Analysis performed per years showed a higher prevalence of serotype 3 Streptococcus pneumoniae during the study period 2004 and 2005. During the entire study period (1999-2006) serotype 3 was most commonly isolated in children older than 24 months (61.2% vs 40.6%;P = 0.05) and showed a lower rate of penicillin non-susceptibility (4.0% vs 18%; P = 0.003). CONCLUSION: Streptococcus pneumoniae serotype 3 is an important pathogen in Costa Rican children with otitis media, especially in children older than 24 months of age (P = 0.05). Most serotype 3 isolates were susceptible to penicillin, cephalosporins, macrolides and quinolones. PMID- 19682368 TI - Association of cetuximab with adverse pulmonary events in cancer patients: a comprehensive review. AB - Compounds derived from biologic sources, or biologicals, are increasingly utilized as therapeutic agents in malignancy. Development of anti-cancer targeted therapies from biologics is increasingly being utilized. Cetuximab, a chimeric monoclonal antibody, is one such anti-cancer targeted therapeutic that has shown efficacy in quelling the rate of patient decline in colorectal, head/neck, and non-small cell lung cancer. However, due to the relatively recent addition of biologic compounds to the therapeutic arsenal, information related to adverse reactions is less well known than those seen in traditional chemotherapeutics. Dermatologic reactions have been demonstrated as the most frequent side effect cited during cetuximab therapy for malignancy; however, other effects may lead to greater morbidity. In general, pulmonary complications of therapeutics can lead to significant morbidity and mortality. The purpose of this review is to compile the various pulmonary side effects seen in patients treated with cetuximab for various malignancies, and to compare the incidence of these adverse reactions to standard therapies. PMID- 19682370 TI - Common polymorphisms within the NR4A3 locus, encoding the orphan nuclear receptor Nor-1, are associated with enhanced beta-cell function in non-diabetic subjects. AB - BACKGROUND: Neuron-derived orphan receptor (Nor) 1, nuclear receptor (Nur) 77, and nuclear receptor-related protein (Nurr) 1 constitute the NR4A family of orphan nuclear receptors which were recently found to modulate hepatic glucose production, insulin signalling in adipocytes, and oxidative metabolism in skeletal muscle. In this study, we assessed whether common genetic variation within the NR4A3 locus, encoding Nor-1, contributes to the development of prediabetic phenotypes, such as glucose intolerance, insulin resistance, or beta cell dysfunction. METHODS: We genotyped 1495 non-diabetic subjects from Southern Germany for the five tagging single nucleotide polymorphisms (SNPs) rs7047636, rs1526267, rs2416879, rs12686676, and rs10819699 (minor allele frequencies >or= 0.05) covering 100% of genetic variation within the NR4A3 locus (with D' = 1.0, r2 >or= 0.9) and assessed their association with metabolic data derived from the fasting state, an oral glucose tolerance test (OGTT), and a hyperinsulinemic euglycemic clamp (subgroup, N = 506). SNPs that revealed consistent associations with prediabetic phenotypes were subsequently genotyped in a second cohort (METSIM Study; Finland; N = 5265) for replication. RESULTS: All five SNPs were in Hardy-Weinberg equilibrium (p >or= 0.7, all). The minor alleles of three SNPs, i.e., rs1526267, rs12686676, and rs10819699, consistently tended to associate with higher insulin release as derived from plasma insulin at 30 min(OGTT), AUCC C-peptide-to-AUC Gluc ratio and the AUC Ins30-to-AUC Gluc30 ratio with rs12686676 reaching the level of significance (p or = the median concentrations (3 microg/L for mothers; 6 microg/L for fathers) compared to combined parents' PBB exposure < the median concentrations (AOR = 1.43, 95% CI: 0.89-2.29), although this did not reach statistical significance. In addition, there was a suggestion of increased odds of a male birth for combined parents' enrollment PCB exposure > or = the median concentrations (6 microg/L for mothers; 8 microg/L for fathers) compared to combined parents' enrollment PCB exposure < the median concentrations (AOR = 1.53, 95% CI: 0.93-2.52). CONCLUSION: This study adds to the body of literature on secondary sex ratio and exposure to environmental contaminants. In this population, combined parental exposure to PBBs or PCBs increased the odds of a male birth. Further research is needed to corroborate these findings and shed light on the biological mechanisms by which these types of chemicals may influence the secondary sex ratio. PMID- 19682391 TI - Palliative care for HIV in the era of antiretroviral therapy availability: perspectives of nurses in Lesotho. AB - BACKGROUND: Southern Africa is disproportionately affected by the HIV/AIDS epidemic. In Lesotho 23% of adults are HIV-positive, and only 26% of those in need are accessing antiretroviral treatment (ART). Consequently, about 18,000 people die from AIDS each year. In this situation, palliative care is needed towards the end of life, but is also recommended throughout the HIV disease trajectory. The World Health Organisation (WHO) has produced the Integrated Management of Adolescent and Adult Illness (IMAI) guidelines, which includes a palliative care guidebook (as well as acute and chronic ART guidebooks). IMAI aims to facilitate the implementation of integrated HIV/AIDS care in resource poor areas. The opinions of health workers towards this integrated approach to care and the use of IMAI has not been considered in previous research studies. This paper therefore aims to address some of these issues. METHODS: Semi structured interviews were conducted with six key informants and ten nurses in Lesotho. The interviews were transcribed verbatim and analysed using content thematic analysis. RESULTS: Many nurses described palliative care as synonymous with chronic care and felt that palliative care is necessary for HIV-positive patients despite the introduction of ART. It was thought that the approach taken should be holistic and integrated throughout the disease trajectory. Pain management was noted to be a particular area of need for palliative care, and it was suggested that this could be improved in Lesotho. The IMAI guidelines were thought to be useful, but knowledge of the palliative care booklet was limited. CONCLUSION: Palliative care remains necessary for HIV despite the increasing availability of ART. However, it is currently significantly lacking in Lesotho and many other sub-Saharan African countries. Greater understanding of palliative care amongst health workers is required, as well as strong political will from the Ministry of Health. The IMAI guidelines are a useful tool for holistic HIV care, including palliative care, but they need to be used more effectively. As ART is becoming increasingly available worldwide, the complex chronic care issues for patients with HIV/AIDS should not be neglected. PMID- 19682392 TI - Intraprostatic Botulinum Toxin Type A injection in patients with benign prostatic enlargement: duration of the effect of a single treatment. AB - BACKGROUND: Botulinum Toxin Type-A (BoNT/A) intraprostatic injection can induce prostatic involution and improve LUTS and urinary flow in patients with Benign Prostatic Enlargement (BPE). However, the duration of these effects is unknown. The objective of this work was to determine the duration of prostate volume reduction after one single intraprostatic injection of 200U of Botulinum Toxin Type-A. METHODS: This is an extension of a 6 month study in which 21 frail elderly patients with refractory urinary retention and unfit for surgery were submitted to intraprostatic injection of BoNT/A-200U, by ultrasound guided transrectal approach. In spite of frail conditions, eleven patients could be followed during 18 months. Prostate volume, total serum PSA, maximal flow rate (Qmax), residual volume (PVR) and IPSS-QoL scores were determined at 1, 3, 6, 12 and 18 months post-treatment. RESULTS: Mean prostate volume at baseline, 82 +/- 16 ml progressively decreased from month one coming to 49 +/- 9,5 ml (p = 0,003) at month six. From this moment on, prostate volume slowly recovered, becoming identical to baseline at 18 months (73 +/- 16 ml, p = 0.03). Albeit non significant, serum PSA showed a 25% decrease from baseline to month 6. The 11 patients resumed spontaneous voiding at month one. Mean Qmax was 11,3 +/- 1,7 ml/sec and remained unchanged during the follow-up period. PVR ranged from 55 +/- 17 to 82 +/- 20 ml and IPSS score from10 to 12 points. CONCLUSION: Intraprostatic BoNT/A injection is safe and can reduce prostate volume for a period of 18 months. During this time a marked symptomatic improvement can be maintained. PMID- 19682393 TI - Romidepsin inhibits Ras-dependent growth transformation of NIH 3T3 fibroblasts and RIE-1 epithelial cells independently of Ras signaling inhibition. AB - BACKGROUND: Despite intensive effort, currently no effective anti-Ras therapies have successfully reached clinical application. Previous studies suggest that the histone deacetylatse (HDAC) inhibitor romidepsin, which is currently in clinical trials for the treatment of multiple malignancies, can block Ras-dependent signaling and growth transformation. These studies suggest that mutational activation of Ras may be a useful biomarker for sensitivity to romidepsin and that the anti-tumor activity of this HDAC inhibitor may involve inhibition of Ras effector-mediated signaling. RESULTS: To rigorously assess romidepsin as an antagonist of Ras, we utilized two well-characterized cell models for Ras transformation. We found that romidepsin blocked the anchorage-dependent and independent growth of NIH 3T3 fibroblasts and RIE-1 epithelial cells transformed by all three Ras isoforms. However, romidepsin treatment also blocked growth transformation caused by other oncoproteins (B-Raf and ErbB2/Neu), suggesting that romidepsin is not selective for Ras. We also observed striking differences in romidepsin-mediated growth inhibition between transformed NIH 3T3 fibroblasts compared to RIE-1 epithelial cells, suggesting that the mechanism by which romidepsin blocks transformation is dependent on cellular context. Finally, we found that romidepsin did not inhibit Ras activation of the ERK and AKT effector pathways in NIH 3T3 and RIE-1 cells, suggesting that romidepsin does not directly antagonize Ras. CONCLUSION: Taken together, our results suggest that romidepsin is not selective for Ras-transformed cells and that the anti-tumor activity of romidepsin is not due to direct inhibition of Ras function. PMID- 19682394 TI - The challenge to detect heart transplant rejection and transplant vasculopathy non-invasively - a pilot study. AB - BACKGROUND: Cardiac allograft rejection and vasculopathy are the main factors limiting long-term survival after heart transplantation.In this pilot study we investigated whether non-invasive methods are beneficial to detect cardiac allograft rejection (Grade 03 R) and cardiac allograft vasculopathy. Thus we compared multi-slice computed tomography and magnetic resonance imaging with invasive methods like coronary angiography and left endomyocardial biopsy. METHODS: 10 asymptomatic long-term survivors after heart transplantation (8 male, 2 female, mean age 52.1 +/- 12 years, 73 +/- 11 months after transplantation) were included. In a blinded fashion, coronary angiography and multi-slice computed tomography and ventricular endomyocardial biopsy and magnetic resonance imaging were compared against each other. RESULTS: Cardiac allograft vasculopathy and atherosclerosis were correctly detected by multi-slice computed tomography and coronary angiography with positive correlation (r = 1). Late contrast enchancement found by magnetic resonance imaging correlated positively (r = 0.92, r2 = 0.85, p < 0.05) with the histological diagnosis of transplant rejection revealed by myocardial biopsy. None of the examined endomyocardial specimen revealed cardiac allograft rejection greater than Grade 1 R. CONCLUSION: A combined non-invasive approach using multi-slice computed tomography and magnetic resonance imaging may help to assess cardiac allograft vasculopathy and cardiac allograft rejection after heart transplantation before applying more invasive methods. PMID- 19682395 TI - Definition of an automated Content-Based Image Retrieval (CBIR) system for the comparison of dermoscopic images of pigmented skin lesions. AB - BACKGROUND: New generations of image-based diagnostic machines are based on digital technologies for data acquisition; consequently, the diffusion of digital archiving systems for diagnostic exams preservation and cataloguing is rapidly increasing. To overcome the limits of current state of art text-based access methods, we have developed a novel content-based search engine for dermoscopic images to support clinical decision making. METHODS: To this end, we have enrolled, from 2004 to 2008, 3415 caucasian patients and collected 24804 dermoscopic images corresponding to 20491 pigmented lesions with known pathology. The images were acquired with a well defined dermoscopy system and stored to disk in 24-bit per pixel TIFF format using interactive software developed in C++, in order to create a digital archive. RESULTS: The analysis system of the images consists in the extraction of the low-level representative features which permits the retrieval of similar images in terms of colour and texture from the archive, by using a hierarchical multi-scale computation of the Bhattacharyya distance of all the database images representation with respect to the representation of user submitted (query). CONCLUSION: The system is able to locate, retrieve and display dermoscopic images similar in appearance to one that is given as a query, using a set of primitive features not related to any specific diagnostic method able to visually characterize the image. Similar search engine could find possible usage in all sectors of diagnostic imaging, or digital signals, which could be supported by the information available in medical archives. PMID- 19682396 TI - Notch signaling is required for maintaining stem-cell features of neuroprogenitor cells derived from human embryonic stem cells. AB - BACKGROUND: Studies have provided important findings about the roles of Notch signaling in neural development. Unfortunately, however, most of these studies have investigated the neural stem cells (NSCs) of mice or other laboratory animals rather than humans, mainly owing to the difficulties associated with obtaining human brain samples. It prompted us to focus on neuroectodermal spheres (NESs) which are derived from human embryonic stem cell (hESC) and densely inhabited by NSCs. We here investigated the role of Notch signaling with the hESC derived NESs. RESULTS: From hESCs, we derived NESs, the in-vitro version of brain derived neurospheres. NES formation was confirmed by increased levels of various NSC marker genes and the emergence of rosette structures in which neuroprogenitors are known to reside. We found that Notch signaling, which maintains stem cell characteristics of in-vivo-derived neuroprogenitors, is active in these hESC-derived NESs, similar to their in-vivo counterpart. Expression levels of Notch signaling molecules such as NICD, DLLs, JAG1, HES1 and HES5 were increased in the NESs. Inhibition of the Notch signaling by a gamma secretase inhibitor reduced rosette structures, expression levels of NSC marker genes and proliferation potential in the NESs, and, if combined with withdrawal of growth factors, triggered differentiation toward neurons. CONCLUSION: Our results indicate that the hESC-derived NESs, which share biochemical features with brain-derived neurospheres, maintain stem cell characteristics mainly through Notch signaling, which suggests that the hESC-derived NESs could be an in vitro model for in-vivo neurogenesis. PMID- 19682397 TI - Vasohibin inhibits angiogenic sprouting in vitro and supports vascular maturation processes in vivo. AB - BACKGROUND: The murine homologue of human vasohibin (mVASH1), a putative antiangiogenic protein, was investigated for its effects on in vitro and in vivo angiogenesis. METHODS: Cell growth and migration were analyzed in murine fibroblasts, smooth muscle cells and endothelial cells. Angiogenic sprouting was studied in human umbilical vein endothelial cells (HUVECs) in the spheroid sprouting assay. In vivo effects on blood vessel formation were investigated in the chorioallantoic membrane (CAM) assay and in the C57BL/6 melanoma xenograft model. RESULTS: Purified murine and human VASH1 protein induced apoptosis of murine fibroblasts in vitro, but not of vascular aortic smooth muscle cells (AoSMC) or endothelial cells. Adenoviral overexpression of murine and human VASH1 inhibited capillary sprouting of HUVECs in the spheroid assay. Administration of recombinant murine and human VASH1 inhibited growth of large vessels in the CAM assay and promoted the formation of a dense, fine vascular network. Murine VASH1 overexpressing B16F10 melanomas displayed a reduction in large vessels and vascular area. Moreover, tumors showed more microvessels that stained positive for the mural cell markers alpha-smooth muscle cell actin (ASMA) and proteoglycan (NG2). CONCLUSION: Our data imply that murine VASH1 causes angiogenic remodelling by inhibiting angiogenic sprouting and large vessel growth, thereby supporting the formation of a vascular bed consisting predominantly of mature microvessels. PMID- 19682398 TI - Influence of environmental factors on food intake and choice of beverage during meals in teenagers: a laboratory study. AB - Environmental conditions influence meal size in adults and children. Intake of sweet drinks could contribute significantly to energy intake and potentially affect body weight, particularly in young individuals. The objectives of the present study were to measure the lunch intake of food and drinks under controlled laboratory settings in teenagers and to compare the influence of different meal conditions. Normal-weight adolescents (fourteen males and fifteen females) participated in four standardised lunches, scheduled 1 week apart. The same popular items (meat dish, dessert, water, juice, soda) were served at all meals. Ad libitum intake was measured under four conditions: subjects ate alone; in groups; alone while viewing television; alone while listening to music. Visual analogue scales were used to assess pre- and post-meal hunger and thirst and meal palatability. Energy, solid food and fluid intake was different (significantly lower) only in the 'eating in group' condition, in spite of identical intensity of pre-meal hunger. More soda was consumed when participants were watching television, and more water was consumed while listening to music. Across all conditions, more soda than water was consumed. Post-meal ratings of hunger, thirst and palatability did not differ between conditions. We concluded that, in teenagers, a 'social inhibition' effect appears rather than the 'social facilitation' previously reported in adults. Although teenagers do not respond to the presence of television or another 'distractor' such as music by eating more, they do ingest more soda when the television is on. The social significance of meals, conditioned responses and habituation to 'distractors' may be different between adolescents and adults. PMID- 19682399 TI - The effect of dairy foods on CHD: a systematic review of prospective cohort studies. AB - There is interest in the degree to which fats in dairy foods contribute to CHD. We undertook a systematic review to investigate the effect of dairy consumption on CHD using prospective cohort studies. A systematic search of electronic databases identified studies relating dairy food intake in adulthood to episodes or death from CHD, IHD and myocardial infarction. Included studies were assessed for quality based on study methodology, validity of dietary assessment, success of follow-up, standardised assessment of CHD, IHD or myocardial infarction end points and appropriateness of statistical adjustment. Data from twelve cohorts involving >280,000 subjects were included. Most studies had follow-up of >80 %, adjusted statistically for three or more confounders and used standard criteria to determine end points. About half the studies used a validated FFQ, administered the FFQ more than once or had follow-up of >20 years. Fewer than half the studies involved subjects representative of the general population. Four of the twelve cohorts found no association between dairy intake and CHD. Eight studies reported varying relationships between different dairy foods and CHD or differential associations based on race, sex or over time. Although dairy foods contribute to the SFA composition of the diet, this systematic review could find no consistent evidence that dairy food consumption is associated with a higher risk of CHD. This could be due to the limited sensitivity of the dietary assessment methods to detect an effect of a single food in a mixed diet on complex clinical outcomes. PMID- 19682400 TI - Diet and deprivation in pregnancy. AB - Deprivation is associated with poor pregnancy outcome but the role of nutrition as a mediating factor is not well understood. We carried out a prospective cohort study of 1461 singleton pregnancies in Aberdeen, UK during 2000-6. We measured nutrient intake and supplement use, B vitamin and homocysteine status, birth weight, gestational age, neonatal treatment and socio-economic deprivation status. Women in the most deprived deciles were approximately 6 years younger and half as likely to take folic acid supplements periconceptually as the least deprived mothers. Deprivation was associated with low blood folate, high homocysteine and diets low in protein, fibre and many of the vitamins and minerals. The diets of the more deprived women were also characterised by low intakes of fruit, vegetables and oily fish and higher intakes of processed meat, fried potatoes, crisps and snacks. Deprivation was related to preterm birth (OR 1.14 (95 % CI 1.03, 1.25); P = 0.009) and whether the baby required neonatal treatment (OR 1.07 (95 % CI 1.01, 1.14); P = 0.028). Low birth weight was more common in women consuming diets low in vitamin C (OR 0.79 (95 % CI 0.64, 0.97); P = 0.028), riboflavin (OR 0.77 (95 % CI 0.63, 0.93); P = 0.008), pantothenic acid (OR 0.79 (95 % CI 0.65, 0.97); P = 0.023) and sugars (OR 0.78 (95 % CI 0.64, 0.96); P = 0.017) even after adjustment for deprivation index, smoking, marital status and parity. Deprivation in pregnancy is associated with diets poor in specific nutrients and poor diet appears to contribute to inequalities in pregnancy outcome. Improving the nutrient intake of disadvantaged women of childbearing age may potentially improve pregnancy outcome. PMID- 19682401 TI - Antioxidant efficacy of curcuminoids from turmeric ( Curcuma longa L.) powder in broiler chickens fed diets containing aflatoxin B1. AB - A 3-week-feeding study (1-21 d post-hatch) was conducted to evaluate the efficacy of total curcuminoids (TCMN), as an antioxidant, to ameliorate the adverse effects of aflatoxin B1 (AFB1) in broiler chickens. Turmeric powder (Curcuma longa L.) that contained 2.55 % TCMN was used as a source of TCMN. Six cage replicates of five chicks each were assigned to each of six dietary treatments, which included: basal diet; basal diet supplemented with 444 mg/kg TCMN; basal diet supplemented with 1.0 mg/kg AFB1; basal diet supplemented with 74 mg/kg TCMN and 1.0 mg/kg AFB1; basal diet supplemented with 222 mg/kg TCMN and 1.0 mg/kg AFB1; basal diet supplemented with 444 mg/kg TCMN and 1.0 mg/kg AFB1. The addition of 74 and 222 mg/kg TCMN to the AFB1 diet significantly (P < 0.05) improved weight gain and feed efficiency. Increase (P < 0.05) in relative liver weight in birds fed AFB1 was significantly reduced (P < 0.05) with the addition of 74, 222 and 444 mg/kg TCMN to the AFB1 diet. The inclusion of 222 mg/kg TCMN ameliorated the adverse effects of AFB1 on serum chemistry in terms of total protein, albumin and gamma-glutamyl transferase activity. The decreased antioxidant functions due to AFB1 were also alleviated by the inclusion of 222 mg/kg TCMN. It is concluded that the addition of 222 mg/kg TCMN to the 1.0 mg/kg AFB1 diet demonstrated maximum antioxidant activity against AFB1. PMID- 19682402 TI - Effects of an onion by-product on bioactivity and safety markers in healthy rats. AB - Onions are excellent sources of bioactive compounds including fructo oligosaccharides (FOS) and polyphenols. An onion by-product was characterised in order to be developed as a potentially bioactive food ingredient. Our main aim was to investigate whether the potential health and safety effects of this onion by-product were shared by either of two derived fractions, an extract containing the onion FOS and polyphenols and a residue fraction containing mainly cell wall materials. We report here on the effects of feeding these products on markers of potential toxicity, protective enzymes and gut environment in healthy rats. Rats were fed during 4 weeks with a diet containing the products or a control feed balanced in carbohydrate. The onion by-product and the extract caused anaemia as expected in rodents for Allium products. No other toxicity was observed, including genotoxicity. Glutathione reductase (GR) and glutathione peroxidase (GPx1) activities in erythrocytes increased when rats were fed with the onion extract. Hepatic gene expression of Gr, Gpx1, catalase, 5-aminolevulinate synthase and NAD(P)H:quinone oxidoreductase was not altered in any group of the onion fed rats. By contrast, gamma-glutamate cysteine ligase catalytic subunit gene expression was upregulated but only in rats given the onion residue. The onion by-products as well as the soluble and insoluble fractions had prebiotic effects as evidenced by decreased pH, increased butyrate production and altered gut microbiota enzyme activities. In conclusion, the onion by-products have no in vivo genotoxicity, may support in vivo antioxidative defence and alter the functionality of the rat gut microbiota. PMID- 19682403 TI - Cystatin C levels in plasma and peripheral blood mononuclear cells among hyperhomocysteinaemic subjects: effect of treatment with B-vitamins. AB - Homocysteine has been related to increased risk of CVD. Matrix degradation and inflammation may be involved in this link between hyperhomocysteinaemia and CVD. Recent studies suggest that cystatin C can modulate matrix degradation and inflammation. The present study measured cystatin C at protein (plasma) and mRNA levels (peripheral blood mononuclear cells (PBMC)) in hyperhomocysteinaemic individuals (n 37, female seven and male thirty, aged 20-70 years) before and after B-vitamin supplementation for 3 months in a randomised, placebo-controlled double-blind trial. In a cross-sectional study, seventeen of the hyperhomocysteinaemic subjects were age- and sex-matched to healthy controls (n 17). Our main findings were: (i) as compared with controls, hyperhomocysteinaemic subjects tended to have higher plasma concentrations of cystatin C and lower mRNA levels of cystatin C in PBMC; (ii) compared with placebo, treatment of hyperhomocysteinaemic individuals with B-vitamins significantly increased plasma levels of cystatin C and mRNA levels of cystatin C in PBMC; (iii) while plasma levels of cystatin C were positively correlated with plasma levels of TNF receptor-1, mRNA levels of cystatin C in PBMC were inversely correlated with this TNF parameter. Taken together, our findings suggest that disturbed cystatin C levels may be a characteristic of hyperhomocysteinaemic individuals, potentially related to low-grade systemic inflammation in hyperhomocysteinaemic subjects, and that B-vitamins may modulate cystatin C levels in these individuals. PMID- 19682404 TI - Body composition by 2H dilution in Gambian infants: comparison with UK infants and evaluation of simple prediction methods. AB - Gambian infants show growth faltering, but the underlying body composition is unknown. The present study aimed to compare body composition in Gambian and UK infants using 2H dilution; and to evaluate accuracy of bioelectrical impedance analysis (BIA) and creatinine excretion for estimating lean mass (LM), using 2H as the reference. Body composition was measured in thirty Gambian infants, aged 3 18 months, using (1) anthropometry, (2) 2H, (3) BIA (equation of Fjeld et al. Pediatr Res (1990), 27, 98-102) and (4) 5 h urinary creatinine excretion. Compared with UK reference data, Gambian infants were light, short and had reduced BMI and skinfolds. The subscapular skinfold standard deviation score (SDS) was greater than the triceps SDS (P < 0.01), indicating central fat preservation. Both LM and fat mass were reduced in Gambian infants, with or without adjustment for length. However, whereas the Gambia-UK difference in LM increased with age, that in fat mass decreased. Average creatinine excretion was similar to that expected (95.5 (sd 23.2) % recovery), but LM estimates showed unacceptable error in individuals. BIA using Fjeld's equation overestimated total body water and LM (P < 0.001), hence a new equation was developed, with standard error of 0.47 kg LM. In conclusion, Gambian infants characterised by growth faltering had LM deficits that increased with age. However, adiposity increased with age, and showed indications of a more central distribution than in the reference infants. A new BIA equation for LM prediction is presented; however, creatinine excretion is not recommended for LM estimation in this population. PMID- 19682405 TI - GPs' awareness of patients' preference for place of death. AB - BACKGROUND: Being able to die in one's place of choice is an indicator of the quality of end-of-life care. GPs may play a key role in exploring and honouring patients' preferences for place of death. AIM: To examine how often GPs are informed about patients' preferred place of death, by whom and for which patients, and to study the expressed preferred place of death and how often patients die at their preferred place. DESIGN OF STUDY: One-year nationwide mortality retrospective study. SETTING: Sentinel Network of GPs in Belgium, 2006. METHOD: GPs' weekly registration of all deaths (patients aged =1 year). RESULTS: A total of 798 non-sudden deaths were reported. GPs were informed of patients' preferred place of death in 46% of cases. GPs obtained this information directly from patients in 63%. GP awareness was positively associated with patients not being hospitalised in the last 3 months of life (odds ratio [OR] = 3.9; 95% confidence interval [CI] = 2.8 to 5.6), involvement of informal caregivers (OR = 3.3; 95% CI = 1.8 to 6.1), use of a multidisciplinary palliative care team (OR = 2.5; 95% CI = 1.8 to 3.5), and with presence of more than seven contacts between GP and patient or family in the last 3 months of life (OR = 3.0; 95% CI = 2.2 to 4.3). In instances where GPs were informed, more than half of patients (58%) preferred to die at home. Overall, 80% of patients died at their preferred place. CONCLUSION: GPs are often unaware of their patients' preference for place of death. However, if GPs are informed, patients often die at their preferred location. Several healthcare characteristics might contribute to this and to a higher level of GP awareness. PMID- 19682406 TI - Suppression of the multidrug transporter P-glycoprotein using RNA interference in cultured rat astrocytes induced by coriaria lactone. AB - The overexpression of the multidrug resistance gene (MDR-1) and its translational product p-glycoprotein (P-gp) may play an important role in pharmacoresistant epilepsy. We established the rat astrocyte model overexpressing P-gp induced by coriaria lactone and successfully nucleofected it with the siRNA-hairpin expression vector pSIREN-shuttle designed to target MDR-1B mRNA. The mRNA expression of MDR-1B gene was mostly knock down by 67.70% (p<0.01). The expression of P-gp in experimental group was significantly lower than that in negative control (p<0.05), and the rhodamine efflux ratio of experimental group (23.08%) was remarkably lower than that of negative control (78.35%, p<0.01). We first employed RNA interfering to the drug resistance reversal of refractory epilepsy and this may provide a new way for refractory epilepsy remedy. PMID- 19682407 TI - Magnetoencephalography of language: new approaches to understanding the cortical organization of Chinese processing. AB - OBJECTIVE: Chinese is a logographic language. Many of its psycholinguistic characteristics differ from those of alphabetic languages. These differences might be expected to entail a different pattern of neural activity underpinning Chinese language processing compared to the processing of alphabetic languages. The aim of the current study was to investigate neural language centers for processing Chinese language information in healthy Chinese speakers using magnetoencephalography (MEG). Overall, we aimed to elucidate language-specific and language-general characteristics of processing across different language scripts. METHODS: Ten healthy Chinese-speaking subjects were asked to silently read genuine Chinese characters and view pseudo-characters in a MEG scanner. The functional language areas were located by overlapping the MEG results over magnetic resonance imaging (MRI) images. RESULTS: Distinctive late magnetic response waves were observed in both hemispheres while the subjects were reading genuine Chinese characters. The polarization of the response waveforms was found to be greater in the left than the right hemisphere. Broca's area was found to be located at the back of gyrus frontalis inferior or gyrus frontalis medius. Wernicke's area was located at gyrus temporalis medius, gyrus temporalis superior and gyrus supramariginalis. In addition, Wernicke's area was activated earlier than Broca's area. CONCLUSION: Native Chinese speakers reading Chinese characters showed neural responses that were lateralized to the left hemisphere. Overall, the functional brain areas activated by reading Chinese in this study corresponded to classical language centers found for alphabetic languages in previous studies, but some differences were also found in the specific patterns of activation. PMID- 19682408 TI - High ICP as trigger of proinflammatory IL-6 cytokine activation in aneurysmal subarachnoid hemorrhage. AB - OBJECTIVE: There is a rising debate about the role of inflammation in the pathogenesis of complications after aneurysmal subarachnoid hemorrhage (SAH) such as intracranial hypertension (intracranial pressure, ICP >20 mmHg). This study aimed to analyse the origin of interleukin-6 (IL-6) in respect to ICP and cerebral metabolism in SAH patients. METHODS: Prospectively, IL-6 was measured in three compartments, the extracellular fluid (ECF) monitored by cerebral microdialysis (MD), cerebrospinal fluid (CSF) and plasma for 10 days after SAH (days 0-4, three times daily; days 5-10, two times daily). Patients were classified having intracranial hypertension (n=7) or normal ICP (n=17) during 10 days after bleeding. Glasgow outcome scale (GOS) was assessed after 3 and 6 months. RESULTS: Patient groups were comparable for age, WFNS and Fisher grade. Intracranial hypertension was associated with an inflammatory response, indicating activation of the inflammatory cascade in the brain (ECF) and systemic circulation with high IL-6 and C-reactive protein (CRP) plasma levels after SAH, the latter associated with unfavourable outcome. The data suggest the ECF but not the CSF as main origin of IL-6 in the systemic circulation in the presence of intracranial hypertension in SAH. DISCUSSION: Intracranial hypertension is associated with a strong activation of the inflammatory cascade in the brain and systemic circulation, and might be underestimated as proinflammmatory trigger in the pathogenesis of complications after SAH. Future therapies targeting anti inflammatory response in plasma may help to reduce the inflammatory cascade responsible for development of intracranial hypertension. PMID- 19682410 TI - Exercise pre-conditioning reduces brain inflammation in stroke via tumor necrosis factor-alpha, extracellular signal-regulated kinase 1/2 and matrix metalloproteinase-9 activity. AB - OBJECTIVE: We sought to determine whether cerebral inflammation in ischemic rats was reduced by a neuroprotective action of pre-ischemic tumor necrosis factor alpha up-regulation, which down-regulated matrix metalloproteinase-9 activity via extracellular signal-regulated kinase 1/2 phosphorylation. MATERIAL AND METHODS: Adult male Sprague-Dawley rats were subjected to 30 minutes of exercise on a treadmill for 3 weeks. Stroke was induced by a 2 hour middle cerebral artery occlusion using an intraluminal filament. The exercised animals were treated with tumor necrosis factor-alpha antibody, UO126 (extracellular signal-regulated kinase 1/2 inhibitor), or both UO126 and doxycycline (matrix metalloproteinase-9 inhibitor). Brain infarct volume was assessed using Nissl staining. Leukocyte infiltration was evaluated using myeloperoxidase immunostaining. Intercellular adhesion molecule-1 and matrix metalloproteinase protein levels were determined by Western blot, and enzyme activity was evaluated using zymography. RESULTS: There was a significant decrease in neurological deficits, brain infarct volume and leukocyte infiltration, in association with reduction in matrix metalloproteinase-9 and intercellular adhesion molecule-1 expression in exercised animals. Exercised animals treated with either tumor necrosis factor-alpha antibody or with UO126 showed a reversal of neurological outcome, infarct volume and leukocyte infiltration. Matrix metalloproteinase-9 activity was reversed, at least partially, but the intercellular adhesion molecule-1 expression was not. Neuroprotection remained when the exercised ischemic rats were treated with both UO126 and doxycycline. CONCLUSION: These results suggest that exercise-induced up regulation of tumor necrosis factor-alpha before stroke and extracellular signal regulated kinase 1/2 phosphorylation play a role in decreasing brain inflammation by regulating matrix metalloproteinase-9 activity. PMID- 19682409 TI - Epileptiform discharges and neuronal plasticity in the acute peri-infarct cortex of rats. AB - While the peri-infarct cortex is thought to be responsible for functional recovery, the site is also a strong candidate for post-stroke seizures. Since it is crucial to identify the conditions when the site is changed with such beneficial or detrimental results, the peri-infarct changes were investigated before and just after inducing a focal infarct on rat cortex. The receptive fields in the peri-infarct cortex began to increase a few hours after the infarct, and reached a statistical significance at 6 hours (Dunnett post hoc tests; p<0.05). In temporal association with these changes, EEG in the peri infarct cortex showed epileptiform activities containing large-amplitude spike and-wave discharges. The gross amplitude, peak-to-peak amplitude and burst frequency showed statistically significant increases within 4 hours, in comparison to those of the controls (Dunnett post hoc tests; p<0.05). FFT power spectrum analyses showed a distinct increase in approximately 25 Hz frequency bands in the post-stroke groups. The homogeneous area of the contralateral hemisphere in the infarct group, in contrast, did not show such plastic or excitability changes. This study demonstrated, for the first time, that the peri infarct cortex acquires the characteristics of potential epileptogenesis and functional recovery within hours of a stroke. PMID- 19682411 TI - Assessment of changes in the epidemiology of Clostridium difficile isolated from diarrheal patients in Hungary. AB - 150 Clostridium difficile strains isolated from diarrheal feces were collected from three parts of Hungary and the presence of genes responsible for toxin A and B, and binary toxin production were examined. MIC distribution against clindamycin, erythromycin, metronidazole, moxifloxacin and rifampin of 80 toxigenic strains selected from the above-mentioned strains and 20 large clostridial toxins (LCTs)-positive strains chosen from our earlier strain collection were determined. 80% of the examined 150 strains were positive for both tcdA and tcdB, and no toxin A-negative, toxin B-positive isolates were found during the study period. 5.3% of toxigenic strains proved to be positive for binary toxin too. Among binary toxin-positive strains, one strain showed the same pattern characteristic of PCR ribotype 027. Comparison of recent findings and our earlier results, the prevalence of toxin-producing and binary toxin-positive strains among C. difficile isolated from diarrheal specimens increased. No metronidazole resistant isolate was detected among strains isolated in 2002-2003 and 2006-2007. The rates of resistance to erythromycin, clindamycin, moxifloxacin and rifampin among strains isolated between 2006 and 2007 were 25%, 27.5%, 25% and 6.3%, respectively. Erythromycin resistance was frequently associated with clindamycin and moxifloxacin resistance, however this resistant phenotype was not found among strains isolated in 2002-2003. PMID- 19682425 TI - New tricks by an old dogma: mechanisms of the Organizational/Activational Hypothesis of steroid-mediated sexual differentiation of brain and behavior. AB - The hormonal regulation of sexual behavior has been the topic of study for over 50 years and yet controversies persist regarding the importance of early versus late events and the identity of the critical neural and cellular substrates. We have taken a mechanistic approach toward the masculinizing actions of the gonadal steroid estradiol, as a means to understand how organization of the neuroarchitechture during a perinatal sensitive period exerts enduring influences on adult behavior. We have identified important roles for prostaglandins, FAK and paxillin, PI3 kinase and glutamate, and determined that cell-to-cell signaling is a critical component of the early organizational events. We have further determined that the mechanisms mediating different components of sexual behavior are distinct and regionally specific. The multitude of mechanisms by which the steroid estradiol, exerts divergent effects on the developing nervous system provides for a multitude of phenotypes which can vary significantly both within and between the sexes. PMID- 19682427 TI - Structure, dynamics and mapping of membrane-binding residues of micelle-bound antimicrobial peptides by natural abundance (13)C NMR spectroscopy. AB - Worldwide bacterial resistance to traditional antibiotics has drawn much research attention to naturally occurring antimicrobial peptides (AMPs) owing to their potential as alternative antimicrobials. Structural studies of AMPs are essential for an in-depth understanding of their activity, mechanism of action, and in guiding peptide design. Two-dimensional solution proton NMR spectroscopy has been the major tool. In this article, we describe the applications of natural abundance (13)C NMR spectroscopy that provides complementary information to 2D (1)H NMR. The correlation of (13)Calpha secondary shifts with both 3D structure and heteronuclear (15)N NOE values indicates that natural abundance carbon chemical shifts are useful probes for backbone structure and dynamics of membrane peptides. Using human LL-37-derived peptides (GF-17, KR-12, and RI-10), as well as amphibian antimicrobial and anticancer peptide aurein 1.2 and its analog LLAA, as models, we show that the cross peak intensity plots of 2D (1)H-(13)Calpha HSQC spectra versus residue number present a wave-like pattern (HSQC wave) where key hydrophobic residues of micelle-bound peptides are located in the troughs with weaker intensities, probably due to fast exchange between the free and bound forms. In all the cases, the identification of aromatic phenylalanines as a key membrane-binding residue is consistent with previous intermolecular Phe-lipid NOE observations. Furthermore, mutation of one of the key hydrophobic residues of KR 12 to Ala significantly reduced the antibacterial activity of the peptide mutants. These results illustrate that natural abundance heteronuclear-correlated NMR spectroscopy can be utilized to probe backbone structure and dynamics, and perhaps to map key membrane-binding residues of peptides in complex with micelles. (1)H-(13)Calpha HSQC wave, along with other NMR waves such as dipolar wave and chemical shift wave, offers novel insights into peptide-membrane interactions from different angles. PMID- 19682428 TI - Rev-erb beta regulates the Srebp-1c promoter and mRNA expression in skeletal muscle cells. AB - The nuclear hormone receptor, Rev-erb beta operates as a transcriptional silencer. We previously demonstrated that exogenous expression of Rev-erb betaDeltaE in skeletal muscle cells increased Srebp-1c mRNA expression. We validated these in vitro observations by injection of an expression vector driving Rev-erb betaDeltaE expression into mouse tibialis muscle that resulted in increased Srebp-1c mRNA expression. Paradoxically, Rev-erb beta siRNA expression in skeletal muscle cells repressed Srebp-1c expression, and indicated that Rev erb beta expression was necessary for Srebp-1c expression. ChIP analysis demonstrated that Rev-erb beta was recruited to the Srebp-1c promoter. Moreover, Rev-erb beta trans-activated the Srebp-1c promoter, in contrast, Rev-erb beta efficiently repressed the Rev-erb alpha promoter, a previously characterized target gene. Finally, treatment with the Rev-erb agonist (hemin) (i) increased the trans-activation of the Srebp-1c promoter by Rev-erb beta; and (ii) increased Rev-erb beta and Srebp-1c mRNA expression. These data suggest that Rev-erb beta has the potential to activate gene expression, and is a positive regulator of Srebp-1c, a regulator of lipogenesis. PMID- 19682426 TI - A label-free mass spectrometry method for the quantification of protein isotypes. AB - Successful quantitative mass spectrometry (MS) requires strategies to link the mass spectrometer response to the analyte abundance, with the response being dependent on more factors than just analyte abundance. Label-dependent strategies rely on the incorporation of an isotopically labeled internal standard into the sample. Current label-free strategies (performed without internal standards) are useful for analyzing samples that are unsuitable for isotopic labeling but are less accurate. Here we describe a label-free technique applicable to analysis of products from related genes (isotypes). This approach enables the invariant tryptic peptide sequences within the family to serve as "built-in" internal standards and the isotype-specific peptide sequences to report the amount of the various isotypes. A process of elimination segregates reliably trypsin-released standard and reporter peptides from unreliably released peptides. The specific MS response factors for these reporter and standard peptides can be determined using synthetic peptides. Analysis of HeLa tubulin digests revealed peptides from betaI , betaII-, betaIII-, betaIVb-, and betaV-tubulin, eight of which were suitable; along with five standard peptides for quantification of the beta-tubulin isotypes. To show the utility of this method, we determined that betaI-tubulin represented 77% and betaIII-tubulin represented 3.2% of the total HeLa beta tubulin. PMID- 19682429 TI - Aggresome-like structure induced by isothiocyanates is novel proteasome-dependent degradation machinery. AB - Unwanted or misfolded proteins are either refolded by chaperones or degraded by the ubiquitin-proteasome system (UPS). When UPS is impaired, misfolded proteins form aggregates, which are transported along microtubules by motor protein dynein towards the juxta-nuclear microtubule-organizing center to form aggresome, a single cellular garbage disposal complex. Because aggresome formation results from proteasome failure, aggresome components are degraded through the autophagy/lysosome pathway. Here we report that small molecule isothiocyanates (ITCs) can induce formation of aggresome-like structure (ALS) through covalent modification of cytoplasmic alpha- and beta-tubulin. The formation of ALS is related to neither proteasome inhibition nor oxidative stress. ITC-induced ALS is a proteasome-dependent assembly for emergent removal of misfolded proteins, suggesting that the cell may have a previously unknown strategy to cope with misfolded proteins. PMID- 19682430 TI - MicroRNA-21 promotes cell proliferation and down-regulates the expression of programmed cell death 4 (PDCD4) in HeLa cervical carcinoma cells. AB - MicroRNAs are involved in cancer-related processes. The microRNA-21(miR-21) has been identified as the only miRNA over-expressed in a wide variety of cancers, including cervical cancer. However, the function of miR-21 is unknown in cervical carcinomas. In this study, we found that the inhibition of miR-21 in HeLa cervical cancer cells caused profound suppression of cell proliferation, and up regulated the expression of the tumor suppressor gene PDCD4. We also provide direct evidence that PDCD4-3'UTR is a functional target of miR-21 and that the 18bp putative target site can function as the sole regulatory element in HeLa cells. These results suggest that miR-21 may play an oncogenic role in the cellular processes of cervical cancer and may serve as a target for effective therapies. PMID- 19682431 TI - Calneuron I inhibits Ca(2+) channel activity in bovine chromaffin cells. AB - Calneuron I (CalnI) is a calmodulin-like protein that contains two functional EF hand motifs at the N-terminal and a hydrophobic segment at the C-terminal. CalnI was cloned from the adult rat cortex and fused with GFP at its N-terminal. When expressed in bovine chromaffin cells, wild-type CalnI was localized at the plasma membrane. However, a mutant that lacked the hydrophobic segment was localized in the cytosol and nucleus, while a Ca(2+)-binding-deficient mutant was found in the cytosol and at the plasma membrane. Evaluation using the whole-cell patch-clamp technique revealed that Ca(2+) currents were inhibited by both wild-type CalnI and the Ca(2+)-binding-deficient mutant. When the bovine N-type Ca(2+) channel was expressed in 293T cells, Ca(2+) currents were mostly inhibited by co expression of CalnI, but not by the mutant without the hydrophobic tail. These results suggest that CalnI attenuates Ca(2+) channel activity and that its subcellular localization is important for this effect. PMID- 19682432 TI - Splicing of intron 3 of human BACE requires the flanking introns 2 and 4. AB - Regulation of proteolytic cleavage of the amyloid precursor protein by the aspartic protease BACE may occur by alternative splicing and the generation of enzymatically inactive forms. In fact, the presence of exonic donor and acceptor sites for intron 3 generates the two deficient variants BACE457 and BACE476. In HEK293 cells, when introns are inserted separately in the BACE cDNA, we found that whilst introns 2 and 4 are efficiently spliced out, intron 3 is not removed. On the other hand, splicing to wild-type BACE is restored when intron 3 is flanked by the two other introns. The presence of all three introns also leads to alternative splicing of intron 3 and the generation of BACE476. In contrast, BACE457 expression takes place only after mutating the donor splice site of intron 3, indicating that additional regulatory elements are necessary for the use of the splicing site within exon 4. Overall, our data demonstrate that a complex splicing of intron 3 regulates the maturation of the BACE mRNA. This appears orchestrated by domains present in the exons and introns flanking intron 3. Excessive BACE activity is a risk factor for Alzheimer's disease, therefore this complex regulation might guarantee low neuronal BACE activity and disease prevention. PMID- 19682433 TI - Constitutive androstane receptor transcriptionally activates human CYP1A1 and CYP1A2 genes through a common regulatory element in the 5'-flanking region. AB - Phenobarbital has long been known to increase cellular levels of CYP1A1 and CYP1A2 possibly through a pathway(s) independent of aryl hydrocarbon receptor. We have investigated the role of constitutive androstane receptor (CAR), a xenobiotic-responsive nuclear receptor, in the transactivation of human CYP1A1 and CYP1A2. These genes are located in a head-to-head orientation, sharing a 5' flanking region. Reporter assays were thus performed with dual-reporter constructs, containing the whole or partially deleted human CYP1A promoter between two different reporter genes. In this system, human CAR (hCAR) enhanced the transcription of both genes through common promoter regions from -461 to -554 and from -18089 to -21975 of CYP1A1. With reporter assays using additional deleted and mutated constructs, electrophoresis mobility shift assays and chromatin immunoprecipitation assays, an ER8 motif (everted repeat separated by eight nucleotides), located at around -520 of CYP1A1, was identified as an hCAR responsive element and a binding motif of hCAR/human retinoid X receptor alpha heterodimer. hCAR enhanced the transcription of both genes also in the presence of an aryl hydrocarbon receptor ligand. Finally, hCAR activation increased CYP1A1 and CYP1A2 mRNA levels in cultured human hepatocytes. Our results indicate that CAR transactivates human CYP1A1 and CYP1A2 in human hepatocytes through the common cis-element ER8. Interestingly, the ER8 motif is highly conserved in the CYP1A1 proximal promoter sequences of various species, suggesting a fundamental role of CAR in the xenobiotic-induced expression of CYP1A1 and CYP1A2 independent of aryl hydrocarbon receptor. PMID- 19682434 TI - Curcumin potentiates the antitumor effects of gemcitabine in an orthotopic model of human bladder cancer through suppression of proliferative and angiogenic biomarkers. AB - Little progress has been made in the last three decades in the treatment of bladder cancer. Novel agents that are nontoxic and can improve the current standard of care of this disease are urgently needed. Curcumin, a component of Curcuma longa (also called turmeric), is one such agent that has been shown to suppress pathways linked to oncogenesis, including cell survival, proliferation, invasion and angiogenesis. We investigated whether curcumin has potential to improve the current therapy for bladder cancer, using an orthotopic mouse model. Curcumin potentiated the apoptotic effects of gemcitabine against human bladder cancer 253JBV cells in culture. Electrophoretic mobility shift assay revealed that curcumin also suppressed the gemcitabine-induced activation of the cell survival transcription factor NF-kappaB. In an orthotopic mouse model, bioluminescence imaging revealed that while curcumin alone significantly reduced the bladder tumor volume, maximum reduction was observed when curcumin was used in combination with gemcitabine (P<0.01 versus vehicle; P<0.01 versus gemcitabine alone). Curcumin also significantly decreased the proliferation marker Ki-67 and microvessel density (CD31) (P<0.01 versus vehicle; P<0.01 versus gemcitabine alone), but maximum reduction occurred when it was combined with gemcitabine (P<0.01 versus vehicle; P<0.01 versus gemcitabine alone). Curcumin abolished the constitutive activation of NF-kappaB in the tumor tissue; induced apoptosis, and decreased cyclin D1, VEGF, COX-2, c-myc and Bcl-2 expression in the bladder cancer tissue. Overall our results suggest that curcumin alone exhibits significant antitumor effects against human bladder cancer and it further potentiates the effects of gemictabine, possibly through the modulation of NF kappaB signaling pathway. PMID- 19682436 TI - Involvement of matrix metalloproteinase-3 in CCL5/CCR5 pathway of chondrosarcomas metastasis. AB - CCL5 (previously called RANTES) was originally recognized as a product of activated T cells, and plays a crucial role in the migration and metastasis of human cancer cells. It has been reported that the effect of CCL5 is mediated via CCR receptors. We found that human chondrosarcoma tissues had significant expression of the CCL5 and CCR5, which was higher than that in normal cartilage. We also found CCL5 increased the migration and matrix metalloproteinases-3 (MMP) 3 expression in human chondrosarcoma cells (JJ012 cells). In addition, MMP-3 small interfering RNA and inhibitor inhibited the CCL5-induced cell migration. Activations of phosphatidylinositol 3-kinase (PI3K), Akt and NF-kappaB pathways after CCL5 treatment was demonstrated, and CCL5-induced expression of MMP-3 and migration activity was inhibited by the specific inhibitor of PI3K, Akt and NF kappaB cascades. Taken together, these results indicate that CCL5 and CCR5 interaction enhanced migration of chondrosarcoma cells through the increase of MMP-3 production. PMID- 19682435 TI - Conformation and recognition of DNA modified by a new antitumor dinuclear PtII complex resistant to decomposition by sulfur nucleophiles. AB - Reported herein is a detailed biochemical and molecular biophysics study of the molecular mechanism of action of antitumor dinuclear Pt(II) complex [{PtCl(DACH)}(2)-mu-Y](4+) [DACH=1,2-diaminocyclohexane, Y=H(2)N(CH(2))(6)NH(2)(CH(2))(2)NH(2)(CH(2))(6)NH(2)] (complex 1). This new, long chain bifunctional dinuclear Pt(II) complex is resistant to metabolic decomposition by sulfur-containing nucleophiles. The results show that DNA adducts of 1 can largely escape repair and yet inhibit very effectively transcription so that they should persist longer than those of conventional cisplatin. Hence, they could trigger a number of downstream cellular effects different from those triggered in cancer cells by DNA adducts of cisplatin. This might lead to the therapeutic effects that could radically improve chemotherapy by platinum complexes. In addition, the findings of the present work make new insights into mechanisms associated with antitumor effects of dinuclear/trinuclear Pt(II) complexes possible. PMID- 19682437 TI - Mitochondria and calcium flux as targets of neuroprotection caused by minocycline in cerebellar granule cells. AB - Minocycline, an antibiotic of the tetracycline family, has attracted considerable interest for its theoretical therapeutic applications in neurodegenerative diseases. However, the mechanism of action underlying its effect remains elusive. Here we have studied the effect of minocycline under excitotoxic conditions. Fluorescence and bioluminescence imaging studies in rat cerebellar granular neuron cultures using fura2/AM and mitochondria-targeted aequorin revealed that minocycline, at concentrations higher than those shown to block inflammation and inflammation-induced neuronal death, inhibited NMDA-induced cytosolic and mitochondrial rises in Ca(2+) concentrations in a reversible manner. Moreover, minocycline added in the course of NMDA stimulation decreased Ca(2+) intracellular levels, but not when induced by depolarization with a high K(+) medium. We also found that minocycline, at the same concentrations, partially depolarized mitochondria by about 5-30 mV, prevented mitochondrial Ca(2+) uptake under conditions of environmental stress, and abrogated NMDA-induced reactive oxygen species (ROS) formation. Consistently, minocycline also abrogates the rise in ROS induced by 75 microM Ca(2+) in isolated brain mitochondria. In search for the mechanism of mitochondrial depolarization, we found that minocycline markedly inhibited state 3 respiration of rat brain mitochondria, although distinctly increased oxygen uptake in state 4. Minocycline inhibited NADH-cytochrome c reductase and cytochrome c oxidase activities, whereas the activity of succinate cytochrome c reductase was not modified, suggesting selective inhibition of complexes I and IV. Finally, minocycline affected activity of voltage-dependent anion channel (VDAC) as determined in the reconstituted system. Taken together, our results indicate that mitochondria are a critical factor in minocycline mediated neuroprotection. PMID- 19682438 TI - Cleaved high molecular weight kininogen, a novel factor in the regulation of matrix metalloproteinases in vascular smooth muscle cells. AB - We previously reported that Brown Norway Katholiek rats, which feature a deficiency of plasma kininogens, develop severe abdominal aortic aneurysm. Increased activity of matrix metalloproteinases (MMPs) in the aortic wall, leading to degradation of extracellular matrix components, is considered to play a crucial role in aneurysm formation. Using an in vitro model of vascular smooth muscle cells (VSMCs), cultured from the rat aorta, we investigated whether the cleaved form of high molecular weight kininogen, designated HKa, affects the expression of MMP-9 and MMP-2 and their tissue inhibitors (TIMPs). Treatment of VSMCs with HKa reduced in a concentration-dependent manner IL-1alpha-induced release of MMP-9 and MMP-2, associated with decreased MMP enzymatic activity levels in conditioned media, as demonstrated by gelatin zymography and fluorescein-labeled gelatin substrate assay, respectively. Real-time PCR revealed that HKa reduced corresponding MMP-9 mRNA levels. Further investigations showed that this effect did not result from a modified rate of MMP-9 mRNA degradation. TIMP-1 mRNA levels, already increased as a result of cytokine-stimulation, were significantly enhanced by HKa. Furthermore, we found elevated basal mRNA expression levels of MMP-2 and TIMP-2 in VSMCs derived from kininogen-deficient Brown Norway Katholiek rats. These results demonstrate for the first time that HKa affects the regulation of MMPs in VSMCs. PMID- 19682440 TI - A cuing study of the N2pc component: an index of attentional deployment to objects rather than spatial locations. AB - Symbolic visual cues indicating the location of an upcoming target are believed to invoke endogenous shifts of attention to cued locations. In the present study, we investigated how visual attention is shifted during such cuing paradigms by recording event-related potentials (ERPs). We focused on a component known to index lateralized shifts of perceptual attention during visual search tasks, known as the N2pc component. The ERP data show that attention was shifted to a cued location in anticipation of a target shape when the location is marked by a placeholder object (Experiments 1 and 2). However, when the possible locations were not marked by placeholder objects, we found no evidence for an anticipatory shift of attention to the cued location (Experiment 3). These findings indicate that the perceptual attention mechanism indexed by the N2pc is deployed to objects and not simply locations in space devoid of object structure. PMID- 19682441 TI - Osthole, a potential antidiabetic agent, alleviates hyperglycemia in db/db mice. AB - Osthole is an agent isolated from Cnidium monnieri (L.) Cusson and Angelica pubescens and has been used to treat several diseases, including metabolic syndromes. To investigate the hypoglycemic effects of osthole in diabetic db/db mice and the underlying mechanisms of these effects by in vitro assay, diabetic db/db mice and cell experiments were utilized to understand its possible effects. Osthole significantly activated both PPARalpha and PPARgamma in a dose-dependent manner based on the results of the transition transfection assay. The activation of PPARalpha and PPARgamma by osthole also resulted in an increase in the expression of PPAR target genes such as PPAR itself, adipose fatty acid-binding protein 2, acyl-CoA synthetases, and carnitine palmitoyltransferase-1A. In vitro results suggested that osthole might be a dual PPARalpha/gamma activator, but its chemical structure differed from that of the thiazolidinedione class of antidiabetic drugs. In addition, osthole markedly activated the AMP-activated protein kinase and its downstream acetyl CoA carboxylase molecules by increasing their phosphorylation levels. Finally, obese diabetic db/db mice were treated with osthole by different administered routes, and osthole was found to markedly reduce blood glucose level. Interestingly, osthole did not reduce the blood insulin or lipid levels, two phenomena that did occur in animals treated with insulin sensitizers like PPAR agonists. These results suggest that osthole can alleviate hyperglycemia and could be potentially developed into a novel drug for treatment of diabetes mellitus. PMID- 19682442 TI - A novel phenotype-based approach for systematically screening antiproliferation metallodrugs. AB - Ruthenium (Ru) derivatives have less toxicity and higher water-solubility than cisplatin, giving them great potential as antitumor metallodrugs. In this study, zebrafish were employed as a whole-organism model to screen new Ru compounds for anti-cell proliferation activity. After soaking fish embryos in cisplatin and five Ru derivatives, [Ru(terpy)(bpy)Cl]Cl, [Ru(terpy)(dppz)OH(2)](ClO(4))(2), [Ru(terpy)(tMen)OH(2)](ClO(4))(2), [Ru(terpy)(Me(4)Phen)OH(2)](ClO(4))(2), and Ru(bpy)(2)Cl(2), only cisplatin and [Ru(terpy)(bpy)Cl]Cl-treated embryos displayed obvious phenotypic effects, such as fin-reduction. After further modification of [Ru(terpy)(bpy)Cl]Cl's main structure and the synthesis of two structurally related compounds, [Ru(terpy)(dcbpyH(2))Cl]Cl and [Ru(terpy)(dmbpy)Cl]Cl, only [Ru(terpy)(dmbpy)Cl]Cl exhibited fin-reduction phenotypes. TUNEL assays combined with immunostaining techniques revealed that treatment with cisplatin, [Ru(terpy)(bpy)Cl]Cl, and [Ru(terpy)(dmbpy)Cl]Cl led proliferating fin mesenchymal cells to undergo apoptosis and consequently caused fin-reduction phenotypes. Furthermore, [Ru(terpy)(bpy)Cl]Cl was able to activate the P53-dependent and independent pathways, and induced human hepatoma cells to undergo apoptosis. In summary, it was concluded that the zebrafish model was effective for the screening of phenotype-based antiproliferation metallodrugs. PMID- 19682439 TI - Therapeutic epilepsy research: from pharmacological rationale to focal adenosine augmentation. AB - Epilepsy is a common seizure disorder affecting approximately 70 million people worldwide. Current pharmacotherapy is neuron-centered, frequently accompanied by intolerable side effects, and fails to be effective in about one third of patients. Therefore, new therapeutic concepts are needed. Recent research suggests an astrocytic basis of epilepsy, presenting the possibility of novel therapeutic targets. In particular, dysfunction of the astrocyte-controlled, endogenous, adenosine-based seizure control system of the brain is implicated in seizure generation. Thus, astrogliosis - a pathological hallmark of the epileptic brain - is associated with upregulation of the adenosine-removing enzyme adenosine kinase (ADK), resulting in focal adenosine deficiency. Both astrogliotic upregulation of ADK in epilepsy and transgenic overexpression of ADK are associated with seizures, and inhibition of ADK prevents seizures in a mouse model of pharmacoresistant epilepsy. These findings link adenosine deficiency with seizures and predict that adenosine augmentation therapies (AATs) will likely be effective in preventing seizures. Given the wide-spread systemic and central side effects of systemically administered AATs, focal AATs (i.e., limited to the astrogliotic lesion) are a necessity. This Commentary will discuss the pharmacological rationale for the development of focal AATs. Additionally, several AAT strategies will be discussed: (1) adenosine released from silk-based brain implants; (2) adenosine released from locally implanted encapsulated cells; (3) adenosine released from stem cell-derived brain implants; and (4) adenosine augmenting gene therapies. Finally, new developments and therapeutic challenges in using focal AATs for epilepsy therapy will critically be evaluated. PMID- 19682443 TI - Novel nitro-oxy derivatives of celecoxib for the regulation of colon cancer cell growth. AB - Celecoxib is a non-steroidal anti-inflammatory drug (NSAID) developed as a selective inhibitor of cyclooxygenase-2 (COX-2). Despite the associated cardiovascular toxicity risk, celecoxib has been found to be effective in reducing cancer risk in animal and human studies. In the present study the antiproliferative activity of novel nitro-oxy-methyl substituted analogues of celecoxib (NO-cel), potentially less cardiotoxic, has been investigated in vitro on human colon cancer cells and compared with action of the parent drug. Moreover, experiments were performed in order to evaluate whether COX-2 pharmacological inhibition may affect beta-catenin/E-cadherin signalling pathway. All the tested analogues of celecoxib exerted a significant antiproliferative activity on COX-2 positive HT-29 human colon cancer cells, being less effective on the COX-2 negative SW-480 human colon cancer cell line. In particular, the analogue displaying two nitro-oxy functions fully mimicked the known inhibitory properties of celecoxib, including inhibition of COX-2, as well as of ERK/MAPK and beta-catenin signalling pathways. Interestingly, the latter compound also elicited a strong reorganization of the beta-catenin/E-cadherin complex, which has been suggested to be relevant for colon carcinogenesis. On these premises, NO cel analogues of celecoxib can represent promising colon cancer chemopreventive agents potentially able to affect colon cancer development. PMID- 19682444 TI - Prophylactic role of arjunolic acid in response to streptozotocin mediated diabetic renal injury: activation of polyol pathway and oxidative stress responsive signaling cascades. AB - Diabetic nephropathy is a common cause for end-stage renal disease. Present study investigated the beneficial role of arjunolic acid (AA) against streptozotocin (STZ) induced diabetic nephropathy in rats. Diabetic renal injury was associated with increased kidney weight to body weight ratio, glomerular area and volume, blood glucose (hyperglycemia), urea nitrogen and serum creatinine. This nephro pathophysiology increased the productions of reactive oxygen species (ROS) and reactive nitrogen species (RNS), enhanced lipid peroxidation, protein carbonylation and decreased intracellular antioxidant defense in the kidney tissue. In addition, hyperglycemia activates polyol pathway by increasing aldose reductase (AR) with a concomitant reduction in Na+-K+-ATPase activity. Investigating the oxidative stress responsive signaling cascades, we found the activation of PKCdelta, PKCvarepsilon, MAPKs and NF-kappaB (p65) in the renal tissue of the diabetic animals. Furthermore, hyperglycemia disturbed the equilibrium between the pro and anti-apoptotic members of Bcl-2 family of proteins as well as reduced mitochondrial membrane potential, elevated the concentration of cytosolic cytochrome C and caspase-3 activity. Treatment of AA effectively ameliorated diabetic renal dysfunctions by reducing oxidative as well as nitrosative stress and deactivating the polyol pathways. Histological studies also support the experimental findings. Results suggest that AA might act as a beneficial agent against the renal dysfunctions developed in STZ-induced diabetes. PMID- 19682445 TI - Detection of antibodies in cardiac autoimmunity. AB - Cardiac autoimmunity has been widely studied over the last years. A lot of cardiac antigens have been identified. Laboratory tests for detection of antibodies against the heart are very helpful in diagnosing and monitoring cardiac autoimmune diseases, but the prevalence of anti-heart antibodies varies a lot between the tests. The aim of this paper is to review the number of laboratory assays used today and to determine the most suitable test for detection of cardiac autoantibodies in human serum. PMID- 19682446 TI - Evidence for an effect of clozapine on the regulation of fat-cell derived factors. AB - OBJECTIVES: The aims of this study were to investigate the effects of clozapine on adipocyte differentiation and its regulation for fat-cell derived factors. MATERIALS AND METHODS: 3T3-L1 preadipocytes were induced into differentiated adipocytes by the addition of 5 microg/ml of insulin, 1 micromol/l dexamethasone, 10 mmol/l IBMX, 1% DMSO, and 10% FBS in DMEM medium. The semi-quantitative RT-PCR was performed to determine the mRNA levels of PPAR gamma, C/EBP alpha, ADD1/SREBP(1C), LPL, and DGAT1. The expression levels of LPL and DGAT1 proteins in the adipocytes treated with clozapine or rosiglitazone were determined by Western blot analysis. The triglyceride concentration was determined by use of GPO-POD assay Kit. RESULTS: Clozapine enhanced the expression level of ADD1/SREBP(1C) mRNA and triglyceride concentration in the differentiated adipocytes. Clozapine significantly suppressed the expression levels of LPL mRNA and LPL protein with a dose-dependent and time-dependent manner, respectively. CONCLUSION: These data suggest that clozapine might play an important role in inducing adipocyte differentiation and the regulation of fat-cell derived factors. PMID- 19682447 TI - AgNORs count and DNA ploidy in liver biopsies from patients with schistosomal liver cirrhosis and hepatocellular carcinoma. AB - OBJECTIVES: Argyrophilic nucleolar organizer regions (AgNOR) proteins are a set of argyrophilic nucleolar proteins that accumulate in highly proliferating cells, whereas their expression is very low in nonproliferating cells. The present study aimed to investigate the potential of DNA flow cytometry (FCM) and AgNORs count in the assessment of cellular kinetics of liver cirrhosis and hepatocellular carcinoma. DESIGN AND METHODS: Small-needle liver biopsies (217) were included and were taken from 84 patients with hepatocellular carcinoma (HCC) (one biopsy from tumor lesion and the other from residual nontumor) liver tissues. Only one biopsy was taken from 49 patients with liver cirrhosis. One part of biopsy was subjected to flow cytometry, and the other, to histopathology and AgNORs counting. RESULTS: An aneuploidy was shown in 44.5% of liver cirrhosis and in 78.6% of tumor sites. Aneuploid HCC cases showed high AgNORs count compared with diploid cases (3.407+/-1.18 vs. 1.74+/-0.9). An extremely significant increase in AgNORs count in tumor lesion (P<0.001) was found compared with residual liver tissues, liver cirrhosis and normal liver (3.89+/-0.827, 1.49+/-0.52, 1.62+/ 0.29, and 1.3+/-0.17, respectively). In liver cirrhosis, dysplasia showed a significant relationship with ploidy (P<0.001) and AgNORs count (P<0.05). CONCLUSION: AgNORs count and DNA ploidy analysis of core biopsy specimens are useful in the assessment of cellular kinetics of liver cirrhosis and hepatocellular carcinoma. PMID- 19682449 TI - Individual variation in baseline and stress-induced corticosterone and prolactin levels predicts parental effort by nesting mourning doves. AB - Endocrine systems have an important mechanistic role in structuring life-history trade-offs. During breeding, individual variation in prolactin (PRL) and corticosterone (CORT) levels affects behavioral and physiological processes that drive trade-offs between reproduction and self-maintenance. We examined patterns in baseline (BL) and stress induced (SI; level following a standard capture restraint protocol) levels of PRL and CORT for breeding mourning doves (Zenaida macroura). We determined whether the relationship of adult condition and parental effort to hormone levels in wild birds was consistent with life-history predictions. Both BL PRL and BL CORT level in adults were positively related to nestling weight at early nestling ages, consistent with the prediction of a positive relationship of hormone levels to current parental effort of adults and associated increased energy demand. Results are consistent with the two hormones acting together at baseline levels to limit negative effects of CORT on reproduction while maintaining beneficial effects such as increased foraging for nestling feeding. Our data did not support predictions that SI responses would vary in response to nestling or adult condition. The magnitude of CORT response in the parents to our capture-restraint protocol was negatively correlated with subsequent parental effort. Average nestling weights for adults with the highest SI CORT response were on average 10-15% lighter than expected for their age in follow-up visits after the stress event. Our results demonstrated a relationship between individual hormone levels and within population variation in parental effort and suggested that hormonal control plays an important role in structuring reproductive decisions for mourning doves. PMID- 19682448 TI - Electrically assisted delivery of macromolecules into the corneal epithelium. AB - Electrically assisted delivery is noninvasive and has been investigated in a number of ocular drug delivery studies. The objectives of this study were to examine the feasibility of electrically assisted delivery of macromolecules such as small interfering RNA (siRNA) into the corneal epithelium, to optimize the iontophoresis and electroporation methods, and to study the mechanisms of corneal iontophoresis for macromolecules. Anodal and cathodal iontophoresis, electroporation and their combinations were the methods examined with mice in vivo. Cyanine 3 (Cy3)-labeled glyceraldehyde-3-phosphate dehydrogenase (GAPDH) siRNA and fluorescein isothiocyanate (FITC)-labeled dextran of different molecular weights (4-70 kDa) were the macromolecules studied. Microscopy and histology after cryostat sectioning were used to analyze and compare the delivery of the macromolecules to the cornea. Iontophoresis was effective in delivering siRNA and dextran up to 70 kDa into the cornea. The electroporation method studied was less effective than that of iontophoresis. Although both iontophoresis and electroporation alone can deliver the macromolecules into the cornea, these methods alone were not as effective as the combination of iontophoresis and electroporation (iontophoresis followed by electroporation). The significant enhancement of dextran delivery in anodal iontophoresis suggests that electroosmosis can be a significant flux-enhancing mechanism during corneal iontophoresis. These results illustrate the feasibility of electrically assisted delivery of macromolecules such as siRNA into the cornea. PMID- 19682450 TI - Similarity in parasite communities of the teleost fish Pinguipes brasilianus in the southwestern Atlantic: infracommunities as a tool to detect geographical patterns. AB - Patterns of distance decay in similarity among communities of the fish Pinguipes brasilianus (Teleostei: Pinguipedidae) from five areas in the southwestern Atlantic were investigated to determine whether the rate of decay varied depending on the community level or the parasite guild analyzed (ectoparasites, adult endoparasites and larval endoparasites). Similarities in species composition were computed at both the component community and infracommunity levels. Similarity indices were calculated between all possible pairs of assemblages from different zones. Infracommunity similarity values between and within host populations were averaged. Significance of linear regressions for similarity values against distance was assessed using randomization tests. Different patterns were observed for each guild, and similarity among infracommunities within host populations varied accordingly. Decay in similarity over distance was recorded for most communities. The slopes differed significantly between infracommunities and component communities in all cases, and stronger decay was always observed for infracommunities. Different geographical patterns in parasite communities were a consequence of variability in parasite availability in the different regions, modulated by oceanographic conditions, as well as variation among species in terms of host specificity and life-cycles strategies. Infracommunities showed a stronger effect of distance than component communities, probably due to the influence of short term and local variability of oceanographic conditions. PMID- 19682451 TI - Purification and expression analysis of imaginal disc growth factor in the silkworm, Bombyx mori. AB - In the present study, we purified and sequenced a homolog of the Drosophila imaginal disc growth factor (IDGF) from the hemolymph of Bombyx mori (BmIDGF). Antibodies against BmIDGF were produced and subsequently used in immunoblotting analyses. The immunoblotting analyses demonstrated an extremely high level of BmIDGF in the hemolymph throughout the period of rapid growth of the organs of B. mori. The results of RT-PCR showed that BmIDGF was predominantly expressed in fat bodies. Real-time RT-PCR revealed that BmIDGF transcripts in fat bodies were highly expressed during the feeding stage but significantly suppressed during the molting, wandering, and pupal stages. Starvation brought about a significant decline of BmIDGF mRNAs in the fat bodies and BmIDGF proteins in the hemolymph. After re-feeding, the BmIDGF transcripts in fat bodies and BmIDGF proteins in the hemolymph increased again. In addition, an immunocytochemical study revealed BmIDGF proteins on the surface of wing discs. The present findings suggest that the level of BmIDGF in the hemolymph was modulated by the fat body in response to nutritional conditions and that BmIDGF was transported to target organs through the hemolymph. PMID- 19682452 TI - Insights to host discrimination and host acceptance behaviour in a parasitoid (Diptera: Asilidae): implications for fitness. AB - The robber fly Mallophora ruficauda is one of the principal pests of apiculture in the Pampas region of Argentina. As adults they prey on honey bees and other insects, while as larvae they are solitary ectoparasitoids of third instar scarab beetle larvae. Females of M. ruficauda lay eggs away from the host in tall grasses. After being dispersed by the wind, larvae drop to the ground, where they dig in search of their hosts. It is known that second instar larvae of M. ruficauda exhibit active host searching behaviour towards its preferred host, third instar larva of Cyclocephala signaticollis. Although the means by which host location occurs has been studied and since superparasitism is a frequent scenario in the field, no information about host discrimination and host acceptance is available. We carried out studies in the field and behavioural experiments in the laboratory to determine if M. ruficauda is capable of quality host discrimination. We also studied if this parasitoid is capable of conspecific detection in order to avoid superparasitism. Finally, we analyzed the conditions under which superparasitism occurs in the field. We report here that the second instar larva of M. ruficauda is able to discriminate the parasitism status of the host by means of chemical cues, but is not capable of detecting conspecifics prior to attacking a host. We also found that the host cannot detect the presence of the parasitoid by means of chemical cues, so that no counter-defense against parasitism occurs. Furthermore, we determined that superparasitism occurs on the heavier hosts, i.e. those with more abundant resources which could harbor several parasitoid individuals. Finally, we discuss the possible implications of larval host location and host discrimination decisions on the fitness of this parasitoid. PMID- 19682453 TI - Cryoprotection in dampwood termites (Termopsidae, Isoptera). AB - In contrast to the majority of the Order, the dampwood termites of the family Termopsidae found in colder regions can experience frost and snow, either in cool temperate areas at high latitudes (45 degrees ), or alpine areas at high elevations (>1000m). This suggests that dampwood termites are adapted to cold climates. We investigated this hypothesis in two dampwood termites, Porotermes adamsoni Froggatt and Stolotermes victoriensis Hill. We measured nest temperatures and atmospheric temperatures of their alpine habitat during winter, and measured survival and recovery at subzero temperatures. We also determined the minimum temperature at which these species remain active and the LT50 values. We used a novel gas chromatographic strategy to examine eight metabolites from individuals of both species collected in winter and summer to identify possible cryoprotectants. Both P. adamsoni and S. victoriensis had significantly higher levels of trehalose, a known cryoprotectant, in winter than in summer; in addition S. victoriensis also had higher levels of unsaturated fatty acid ligands in winter than in summer, consistent with patterns observed for cold adaptation in other organisms. These results are the first to reveal that dampwood termites are adapted to cold climates and use trehalose and unsaturated lipids as cryoprotectants. PMID- 19682454 TI - Tradeoffs between metabolic rate and spiracular conductance in discontinuous gas exchange of Samia cynthia (Lepidoptera, Saturniidae). AB - The insect tracheal system is a unique respiratory system, designed for maximum oxygen delivery at high metabolic demands, e.g. during activity and at high ambient temperatures. Therefore, large safety margins are required for tracheal and spiracular conductance. Spiracles are the entry to the tracheal system and play an important role in controlling discontinuous gas exchange (DGC) between tracheal system and atmosphere in moth pupae. We investigated the effect of modulated metabolic rate (by changing ambient temperature) and modulated spiracular conductance (by blocking all except one spiracles) on gas exchange patterns in Samia pupae. Both, spiracle blocking and metabolic rates, affected respiratory behavior in Samia cynthia pupae. While animals showed discontinuous gas exchange cycles at lower temperatures with unblocked spiracles, the respiratory patterns were cyclic at higher temperatures, with partly blocked spiracles or a combination of these two factors. The threshold for the transition from a discontinuous (DGC) to a cyclic gas exchange ((cyc)GE) was significantly higher in animals with unblocked spiracles (18.7 nmol g(-1) min(-1) vs. 7.9 nmol g(-1) min(-1)). These findings indicate an important influence of spiracle conductance on the DGC, which may occur mostly in insects showing high spiracular conductances and low metabolic rates. PMID- 19682455 TI - Structural alterations in the male reproductive system of the freshwater crayfish, Cherax quadricarinatus (Decapoda, Parastacidae). AB - No diseases affecting reproductive performance have been previously reported in freshwater crayfishes. This study aims to characterise one reproductive system abnormality found in males of Cherax quadricarinatus reared in captivity. Fifteen adult males of C. quadricarinatus (70-110 g) were purchased from San Mateo S.A. farm (Entre Rios, Argentina) each season during 2007. Macroscopic analysis showed that 26.6% of the animals sacrificed in winter presented brownish distal vasa deferentia. Histological analysis showed different levels of structural abnormality in the epithelium of the vasa deferentia and spermatophore. Granular and hyaline haemocytes were identified within the vasa deferentia but no significant differences were found in the sperm count between normal and brownish vas deferens. Histological analysis of the crayfishes sacrificed in autumn also showed these modifications in 22% of the animals, however, they did not show the brownish colour under macroscopic analysis. The similarities between the male reproductive system syndrome in shrimps and the abnormalities found in C. quadricarinatus are notable. An unspecific response to thermic stress is a possible explanation of these structural alterations. PMID- 19682456 TI - A new rod-shaped virus from parasitic wasp Diachasmimorpha longicaudata (Hymenoptera: Braconidae). AB - A new rod-shaped nucleocapsids (NCs) was found inadvertently in Diachasmimorpha longicaudata accessory gland filaments (AGFs). The NCs were 30 nm in diameter and nearly 900 nm in length. They replicated in a small cell type of the AGFs in D. longicaudata, and following oviposition, invaded and proliferated in the hemocytes of a parasitized host Bactrocera dorsalis Hendel. This finding of a completely new virus in the AGF indicate that different geographical populations (subspecies) of D. longicaudata may carry different sybionts. This is the first report showing that the same wasp species, but from a different geographical populations, can carry an entirely different virus. PMID- 19682457 TI - PCR-RFLP analysis of chitinase genes enables efficient genotyping of Metarhizium anisopliae var. anisopliae. AB - A new genotyping tool has been developed and evaluated for Metarhizium anisopliae var. anisopliae. The tool is based on Restriction Fragment Length Polymorphism (RFLP) analysis of three chitinase genes that are functionally linked to insect pathogenicity of this fungus. It allowed for discrimination of 14 genotypes among 22 M. anisopliae var. anisopliae strains of a world wide collection. Analyses revealed that the approach may also be applicable to other Metarhizium varieties. The new tool will be useful for genetic characterization of M. anisopliae var. anisopliae strains, and it is applicable for laboratories with limited access to molecular diagnostic equipment. PMID- 19682458 TI - Characterization of Xenorhabdus isolates from La Rioja (Northern Spain) and virulence with and without their symbiotic entomopathogenic nematodes (Nematoda: Steinernematidae). AB - Eighteen Xenorhabdus isolates associated with Spanish entomopathogenic nematodes of the genus Steinernema were characterized using a polyphasic approach including phenotypic and molecular methods. Two isolates were classified as Xenorhabdus nematophila and were associated with Steinernema carpocapsae. Sixteen isolates were classified as Xenorhabdus bovienii, of which fifteen were associated with Steinernema feltiae and one with Steinernema kraussei. Two X. bovienii Phase II were also isolated, one instable phase isolated from S. feltiae strain Rioja and one stable phase from S. feltiae strain BZ. Four representative bacterial isolates were chosen to study their pathogenicity against Spodoptera littoralis with and without the presence of their nematode host. The four bacterial isolates were pathogenic for S. littoralis leading to septicemia 24h post-injection and killing around 90% of the insect larvae 36 h post-injection, except for that isolated from S. kraussei. After 48 h of injection, this latter isolate showed a lower final population in the larval hemolymph (10(7) instead of 10(8)CFU per larvae) and a lower larval mortality (70% instead of 95-100%). The virulence of the nematode-bacteria complexes against S. littoralis showed similar traits with a significant insect larvae mortality (80-90%) 5 days post-infection except for S. kraussei, although this strain reached similar of larval mortality at 7 days after infection. PMID- 19682459 TI - Invasive Bombus terrestris (Hymenoptera: Apidae) parasitized by a flagellate (Euglenozoa: Kinetoplastea) and a neogregarine (Apicomplexa: Neogregarinorida). AB - The flagellate Crithidia bombi and the neogregarine Apicystis bombi have been found in individuals of Bombus terrestris, a Palaearctic species of bumble bee commercially reared and shipped worldwide for pollination services. B. terrestris has recently entered into the northwestern Patagonia region of Argentina from Chile, where it was introduced in 1998. Prevalence was 21.6% for C. bombi and 3.6% for A. bombi (n=111). The pathogens were not detected in 441 bumble bees belonging to five of the eight known Argentine native species (Bombus atratus, Bombus morio, Bombus bellicosus, Bombus opifex, Bombus tucumanus) collected elsewhere in the country. Although the absence of natural occurrence of C. bombi and A. bombi in Argentine native bumble bees cannot be ascertained at present due to the limited surveys performed, it is important to report their detection in invasive B. terrestris. The invasion event is relatively recent and the accompanying pathogens are not species specific within the genus Bombus. PMID- 19682460 TI - Intraguild interactions between the entomopathogenic fungus Pandora neoaphidis and an aphid predator and parasitoid at the population scale. AB - The interactions that occur between the entomopathogenic fungus Pandora neoaphidis and a predator (Coccinella septempunctata) and a parasitoid (Aphidius ervi) were assessed in microcosm and polytunnel experiments. Transmission of P. neoaphidis to the pea aphid, Acyrthosiphon pisum, was enhanced in the presence of both C. septempunctata and A. ervi in microcosm experiments done under fixed abiotic conditions. In contrast, the reproductive success of A. ervi was reduced in the presence of P. neoaphidis. Despite the increased fungal transmission in the presence of C. septempunctata, there was no additional decrease in the aphid population indicating that P. neoaphidis is functionally redundant in the presence of the coccinellid. In polytunnel experiments the reproductive success of A. ervi was not affected by P. neoaphidis. These results do not support those of the microcosm and may be due to the more natural abiotic conditions in the polytunnel reducing the competitive advantage of the fungus. Microcosms therefore provide an arena in which the interactions between fungal pathogens and other aphid-natural enemies can be assessed however, further assessments at increased spatial scales under more natural abiotic conditions are also required to accurately determine the outcome of these interactions. PMID- 19682462 TI - Describing long-range patterns in leaf vasculature by metaphoric fields. AB - Some patterns in dicotyledonous leaf vasculature depict rather precise, long range structural features. This work identifies and quantifies these previously unrecognized features in terms of an empirically derived mathematical formalism that generates wave-like spatial patterns referred to as metaphoric fields. These patterns were used to specify regularities in the long-range structure of dicot leaf vasculature, and were found to account significantly for the predominant features of all 27 dicot species studied. The conserved features of these metaphoric fields are discussed in terms of existing models for leaf pattern formation based on efflux-protein mediated auxin transport in a developing cellular field. This work highlights the complex, regular, long-range structures existing in leaf vascular patterns, and provides a means for specifying and identifying the inherent global features of vascular patterns which must be accounted for in functional developmental models. PMID- 19682461 TI - Intracardiac renin-angiotensin system and myocardial repair/remodeling following infarction. AB - The circulating renin-angiotensin system (RAS) is a classic endocrine system that regulates cardiovascular homeostasis during physiologic and pathologic states. Accumulated evidence has shown the presence of components of RAS in various tissues, which are upregulated in certain pathological conditions. Locally produced angiotensin (Ang)II may play an important role in tissue repair/remodeling in autocrine and/or paracrine manners. Following acute myocardial infarction (MI), cardiac repair occurs in the infarcted myocardium and structural remodeling is developed in noninfarcted myocardium, which are accompanied by activated cardiac RAS. In this review, the current understanding of independent activation of cardiac RAS and its regulation in the pathogenesis of myocardial repair/remodeling after MI is discussed. PMID- 19682463 TI - Absence of adenosine A1 receptors unmasks pulses of insulin release and prolongs those of glucagon and somatostatin. AB - AIMS: Extracellular ATP modulates pulsatile release of insulin, glucagon and somatostatin by activating P2Y(1) receptors. The present study examines if adenosine via A(1) receptors (A(1)R) interferes with pulsatile islet hormone release. MAIN METHODS: Pancreas was perfused in mice expressing or lacking the A(1) receptor and the hormones measured with radioimmunoassay. Cytoplasmic Ca(2+) was recorded in isolated beta-cells using the fura-2 indicator. KEY FINDINGS: Addition of 10 microM adenosine removed the Ca(2+) transients supposed to coordinate the insulin release pulses. This effect of adenosine was counteracted by 100 nM of the A(1)R antagonist DPCPX. In situ perfusion of the pancreas indicated two phases of islet hormone release when glucose was raised from 3.3 to 16.7 mM. The first phase was characterized by a brief dip followed by a peak, which was more pronounced for insulin and somatostatin than for glucagon. The second phase was markedly affected by knock out of A(1)R. The wild-type A(1)R (+/+) mice, usually lacked statistically verified insulin pulses but generated antisynchronous glucagon and somatostatin pulses with half-widths of 4 min. In the A(1)R (-/-) mice time-average release of insulin during the second phase was almost three times higher than in the controls and 30% of the hormone was released as distinct pulses with half-widths of 3 min. The absence of the A(1)R receptor resulted in 50% prolongation of the pulse cycles of glucagon and somatostatin and loss of their antisynchronous relationship. SIGNIFICANCE: The A(1)R receptor is important both for the amplitude (insulin) and duration (glucagon and somatostatin) of islet hormone pulses. PMID- 19682464 TI - A mouse model of yellow fluorescent protein (YFP) expression in hematopoietic cells to assess leukocyte-endothelial interactions in the microcirculation. AB - In this study, we describe the use of intravital microscopy in a transgenic mouse model expressing yellow fluorescent protein (YFP) under the control of a monocyte specific promoter c-fms (CD115) to track and quantify specific leukocyte subsets. Flow cytometry on peripheral and bone marrow leukocytes revealed that YFP was predominantly expressed by CD11a(+), CD11b(+), and CD14(+) monocytes. In the bone marrow, 67+/-4% of Ly6C(high) F4/80(+) cells were YFP(high) while 55+/-1% of Ly6C(low) F4/80(+) cells were YFP(low) supporting the use of c-fms(YFP) expression as a marker of monocyte lineage. 70+/-7% of CD11b(+) F4/80(+) Ly6C(+) ("triple positive") cells expressed YFP. To assess leukocyte-endothelial interactions in YFP(+) cells in c-fms(YFP+) mice, we evaluated leukocyte adhesion, rolling and local shear stress responses in the cremasteric endothelium 4 h following administration of TNFalpha. TNFalpha resulted in a five-fold increase in adhesion of YFP(+) cells to the endothelium and provided superior discriminative ability in assessing rolling and adhesion events when compared with bright field microscopy. Additionally, when compared with Rhodamine-6G labeled leukocytes or GFP(+) cells in mice transplanted with green fluorescent protein (GFP) positive bone marrow, the level of detail observed in the c fms(YFP+) was greater, with both GFP(+) and YFP(+) cells demonstrating superior signal to noise compared to bright field microscopy. A weak positive linear correlation between wall shear stress and YFP(+) cell adhesion (r(2)=0.20, p<0.05) was seen in the cremasteric microcirculation. Taken together, these data demonstrate the use of c-fms(YFP+) mice in identifying distinct monocyte subsets and highlight the potential of this model for real-time monocyte-endothelial interactions using intravital microscopy. PMID- 19682466 TI - Mechanism of elimination of phosphorylated histone H2AX from chromatin after repair of DNA double-strand breaks. AB - Covalent modifications of histones in chromatin play an important role in regulation of eukaryotic gene expression and DNA repair. Formation of double strand breaks (DSBs) in DNA is followed by the rapid local phosphorylation of the C-terminal serine in the replacement histone H2AX in megabase chromatin domains around DSBs and formation of discrete nuclear foci called gammaH2AX foci. This epigenetic modification of chromatin represents the "histone code" for DNA damage signaling and repair and has been extensively studied during last decade. It is known that after DSB rejoining gammaH2AX foci are eliminated from the nucleus, but molecular mechanism of this elimination remains to be established. However, gammaH2AX elimination can serve as a useful marker of DSB repair in normal cells and tissues. In this paper the available data on kinetics and possible mechanisms of gammaH2AX elimination are reviewed. PMID- 19682465 TI - DNA polymerases at the eukaryotic fork-20 years later. AB - Function of the eukaryotic genome depends on efficient and accurate replication of anti-parallel DNA strands. Eukaryotic DNA polymerases have different properties adapted to perform a wide spectrum of DNA transactions. Here we focus on major players in the bulk replication, DNA polymerases of the B-family. We review the organization of the replication fork in eukaryotes in a historical perspective, analyze contemporary models and propose a new integrative model of the fork. PMID- 19682467 TI - Regulation of damage recognition in mammalian global genomic nucleotide excision repair. AB - Nucleotide excision repair operating throughout the mammalian genome plays a crucial role in the suppression of mutagenesis and carcinogenesis, which can arise from DNA lesions induced by a wide variety of genotoxic agents, such as ultraviolet light and chemical compounds. A key process of this DNA repair pathway, damage recognition, is accomplished through multiple steps including concerted actions of the damaged DNA binding factors XPC and UV-DDB, both of which are implicated in a human cancer-prone genetic disorder, xeroderma pigmentosum. Accumulating evidence indicates that the expression and functions of these damage recognition factors are subject to exquisite regulation at diverse levels, including transcriptional activation, post-translational modification, complex formation, and protein degradation through ubiquitination. PMID- 19682469 TI - Positron emission tomography neuroimaging for a better understanding of the biology of ADHD. AB - Attention-deficit/hyperactivity disorder (ADHD) is a neurobehavioral disorder characterized by inappropriate symptoms of inattention, impulsivity and motor restlessness. Converging data from neuropsychological, genetic, neurochemical and pharmacological studies have implicated fronto-striatal network abnormalities as the likely cause of ADHD. The functional imaging field has evolved rapidly providing unprecedented tools to examine questions regarding the pathophysiology of ADHD and the biological effects of medications used to treat it. Positron emission tomography (PET) provides unique quantitative information on the spatial resolution of radiolabelled molecules in the brain of patients or healthy subjects allowing the longitudinal assessment of physiological parameters such as binding potential over extended periods of time. The main goal of this review is to provide an overview of PET studies performed in ADHD patients, discuss their relative strengths and weaknesses and show how they can complement one another to enable a better understanding of the neurobiology and the neuropharmacology of this disease. PMID- 19682470 TI - Attentional and sensory effects of lowered levels of intrinsic alertness. AB - Low levels of intrinsic alertness are associated with lateralized performance in visual tasks, similar to neglect of the left (ipsilesional) visual hemi-field. However, it is unclear whether reduced alertness produces a specific lateralization of spatial-attentional processes in terms of the prioritization of right- over left-side stimuli, or whether it affects more basic functions of visuo-sensory coding, and/or higher function of the top-down control of selection, of stimuli on the left side. To decide between these alternatives, the present study examined the effects of lowered alertness, induced by a 50-min vigilance task, in a partial-report paradigm of briefly presented letter displays. With only one (unilateral) stimulus in display, no specific hemi-field effects were found under low-alertness conditions, indicating that reduced alertness impairs neither sensory effectiveness nor the top-town control of selection. However, with dual, bilateral stimuli, report accuracy was specifically affected for left-side targets (in subjects who showed comparable performance for both sides under normal-alertness conditions). This pattern can be interpreted in terms of a specific bias in spatial-attentional weighting, where prioritization of stimuli on the right leads to (mild) extinction of targets on the left. Moreover, participants who had a lower general level of alertness also showed a more pronounced re-distribution of weights, evidenced by a more severe imbalance in report accuracy, in a low compared to a normal state of alertness. This suggests that a low general level of intrinsic alertness engenders a specific vulnerability to neglect-like performance with a (mild) left side extinction. PMID- 19682468 TI - Differences between human slow N-acetyltransferase 2 alleles in levels of 4 aminobiphenyl-induced DNA adducts and mutations. AB - Aromatic amines such as 4-aminobiphenyl (ABP) require biotransformation to exert their carcinogenic effects. Genetic polymorphisms in biotransformation enzymes such as N-acetyltransferase 2 (NAT2) may modify cancer risk following exposure. Nucleotide excision repair-deficient Chinese hamster ovary (CHO) cells stably transfected with human cytochrome P4501A1 (CYP1A1) and a single copy of either NAT2*4 (rapid acetylator), NAT2*5B (common Caucasian slow acetylator), or NAT2*7B (common Asian slow acetylator) alleles (haplotypes) were treated with ABP to test the effect of NAT2 polymorphisms on DNA adduct formation and mutagenesis. ABP N acetyltransferase catalytic activities were detectable only in cell lines transfected with NAT2 and were highest in cells transfected with NAT2*4, lower in cells transfected with NAT2*7B, and lowest in cells transfected with NAT2*5B. Following ABP treatment, N-(deoxyguanosin-8-yl)-4-aminobiphenyl (dG-C8-ABP) was the primary adduct formed. Cells transfected with both CYP1A1 and NAT2*4 showed the highest concentration-dependent cytotoxicity, hypoxanthine phosphoribosyl transferase (hprt) mutants, and dG-C8-ABP adducts. Cells transfected with CYP1A1 and NAT2*7B showed lower levels of cytotoxicity, hprt mutagenesis, and dG-C8-ABP adducts. Cells transfected with CYP1A1 only or cells transfected with both CYP1A1 and NAT2*5B did not induce cytotoxicity, hprt mutagenesis or dG-C8-ABP adducts. ABP-DNA adduct levels correlated very highly (r>0.96) with ABP-induced hprt mutant levels following each treatment. The results of the present study suggest that investigations of NAT2 genotype or phenotype associations with disease or toxicity could be more precise and reproducible if heterogeneity within the "slow" NAT2 acetylator phenotype is considered and incorporated into the study design. PMID- 19682471 TI - Attention and material-specific memory in children with lateralized epilepsy. AB - Epilepsy is frequently associated with attention and memory problems. In adults, lateralization of seizure focus impacts the type of memory affected (left-sided lesions primarily impact verbal memory, while right-sided lesions primarily impact visual memory), but the relationship between seizure focus and the nature of the memory impairment is less clear in children. The current study examines the correlation between parent-reported attention problems and material-specific memory (verbal or visual-spatial) in 65 children (ages 6-16) with medically intractable lateralized epilepsy. There were no significant differences in attention and memory between those with left-lateralized epilepsy (n=25) and those with right-lateralized epilepsy (n=40). However, in the left-lateralized group attention problems were significantly negatively correlated only with delayed visual memory (r=-.450, p<.05), while the right-lateralized group demonstrated the opposite pattern (attention problems significantly negatively correlated with delayed verbal memory; r=-.331, p<.05). These findings suggest that lateralization of seizure focus may in fact impact children's memory in a material-specific manner, while problems with attention may impact memory more globally. Therefore, interventions designed to improve attention in children with epilepsy may have utility in improving certain aspects of memory, but further suggest that in children with lateralized epilepsy, material-specific memory deficits may not resolve with such interventions. PMID- 19682472 TI - Touch and feel? Using the rubber hand paradigm to investigate self-touch enhancement in right-hemisphere stroke patients. AB - Following stroke, a patient may fail to report touch administered by another person but claim that she feels touch when it is self-administered. We investigated three explanations for self-touch enhancement: (1) proprioceptive information from the administering hand, (2) attentional modulation, and (3) temporal expectation. Tactile sensation was assessed with vision precluded, and with the affected hand positioned in the left and right hemispace. In four of six experiments, the somatic rubber hand paradigm was used: the Examiner administered stimulation to the patient's affected left hand while guiding the patient's right hand to administer synchronous stimulation to a prosthetic hand. Even though the patient's two hands were not in contact, patients detected the same number of stimulations as when they touched their own hand directly (self-administered condition). Moreover, there was no decline in rates of detection when potentially informative movements of the administering hand were restricted. This demonstrates that patients feel rather than infer stimulation under conditions of self-touch. When patients received stimulation to the affected hand in the opposite hemispace to the hand administering touch to the prosthetic hand, all but one showed self-touch enhancement. Thus, neither proprioceptive information nor attentional modulation at the spatial region of the administering hand provided a sufficient explanation for self-touch enhancement. A follow-up experiment indicated an important role for temporal expectation: a delay, between the patient's stimulation of the prosthetic hand and the Examiner's stimulation of the patient's affected hand, eliminated the self-touch enhancement effect. PMID- 19682473 TI - Lateralized temporal order judgement in dyslexia. AB - Temporal and spatial attentional deficits in dyslexia were investigated using a lateralized visual temporal order judgment (TOJ) paradigm that allowed both sensitivity to temporal order and spatial attentional bias to be measured. Findings indicate that adult participants with a positive screen for dyslexia were significantly less sensitive to the temporal order of the stimuli than control participants, but did not show a significantly different lateral bias. However, the data indicated that performance on the TOJ task dissociated into at least three factors. One loaded on trials with long Stimulus Onset Asynchronies (SOA) and was strongly correlated with full-scale IQ (FSIQ), and, while also correlated with both poor reading and with symptoms of attentional deficit disorder, was not specific to these. The second factor loaded on trials with short SOAs in which the left stimulus was presented first. Low scores on this factor were associated with poor non-word reading accuracy, and factor scores accounted for variance in non-word reading accuracy that was not accounted for by either FSIQ or the presence of a phonological deficit. This suggests that a "left mini-neglect" syndrome, also reported in attention deficit and hyperactivity disorder, may directly contribute to poor non-word reading. However, attentional deficit symptoms loaded not only on this factor, but also on a third factor, representing on trials at short SOAs in which the first stimulus was presented in left hemifield. This suggests that attentional deficit symptoms impaired temporal processing at short SOAs, regardless of the hemifield in with the stimuli were presented. We conclude that people with attentional deficits find a visual TOJ task difficult when the stimuli are presented rapidly, regardless of FSIQ, and that where a rightward attentional bias is present, non-word reading accuracy may be directly impaired, even in the absence of a phonological deficit. PMID- 19682474 TI - Preference for full-fat over low-fat foods among individuals suffering from coronary heart disease and healthy controls. AB - Dietary behaviour can modify the risks for coronary heart disease (CHD). Dietary fat contributes to the sensory characteristics of many foods, but there are individual differences in liking for regular and reduced fat products. Preference for dietary fat might differ between healthy individuals and those of diagnosed CHD status. Preference for full-fat versus low-fat foods was assessed in a group suffering from CHD (N=24) and a healthy control group (N=41). Preferences were evaluated using a series of paired preference tests including 34 food pairs. Further, patterns across fat preference and a range of known risk factors for CHD were examined. The study was conducted in Sweden; March 2004 until May 2006. Overall the full-fat food items were preferred to the low-fat alternatives by both patients and control participants. The pattern of preference responses diverged significantly between patients and controls on only four of the 34 food items, in both directions, why preference related to total fat content per se is not confirmed as differentiating significantly between a group diagnosed with CHD compared to a healthy control group. PMID- 19682475 TI - Central-place seed foraging and vegetation patterns. AB - We investigate how central-place seed foragers with a nest in the proximity of one or more seed sources determine the formation of different vegetation patterns. In particular, we discuss the ecological conditions that lead to the formation of hump-shaped (Janzen-Connell) patterns in a two-dimensional landscape. Our analysis shows that central-place predation can generate Janzen Connell patterns even if predators' movement strategies are exclusively based on resource abundance, both in the single-plant/single-nest case and in a forest with several seed sources. We also show that social foraging may either promote or work against the formation of Janzen-Connell patterns, depending upon the way foragers take advantage of social interactions. PMID- 19682476 TI - Diabetes increases susceptibility of primary cultures of rat proximal tubular cells to chemically induced injury. AB - Diabetic nephropathy is characterized by increased oxidative stress and mitochondrial dysfunction. In the present study, we prepared primary cultures of proximal tubular (PT) cells from diabetic rats 30 days after an ip injection of streptozotocin and compared their susceptibility to oxidants (tert-butyl hydroperoxide, methyl vinyl ketone) and a mitochondrial toxicant (antimycin A) with that of PT cells isolated from age-matched control rats, to test the hypothesis that PT cells from diabetic rats exhibit more cellular and mitochondrial injury than those from control rats when exposed to these toxicants. PT cells from diabetic rats exhibited higher basal levels of reactive oxygen species (ROS) and higher mitochondrial membrane potential, demonstrating that the PT cells maintain the diabetic phenotype in primary culture. Incubation with either the oxidants or mitochondrial toxicant resulted in greater necrotic and apoptotic cell death, greater evidence of morphological damage, greater increases in ROS, and greater decreases in mitochondrial membrane potential in PT cells from diabetic rats than in those from control rats. Pretreatment with either the antioxidant N-acetyl-l-cysteine or a catalase mimetic provided equivalent protection of PT cells from both diabetic and control rats. Despite the greater susceptibility to oxidative and mitochondrial injury, both cytoplasmic and mitochondrial glutathione concentrations were markedly higher in PT cells from diabetic rats, suggesting an upregulation of antioxidant processes in diabetic kidney. These results support the hypothesis that primary cultures of PT cells from diabetic rats are a valid model in which to study renal cellular function in the diabetic state. PMID- 19682477 TI - Inhalation exposure to sulfur mustard in the guinea pig model: clinical, biochemical and histopathological characterization of respiratory injuries. AB - Guinea pigs (GP) were exposed (head only) in individual plethysmographs to various concentrations of sulfur mustard vapor, determined online, using FTIR attached to flow chamber. The LCt(50) and the inhaled LD(50) were calculated at different time points post exposure. Surviving animals were monitored for clinical symptoms, respiratory parameters and body weight changes for up to 30 days. Clinical symptoms were noted at 3 h post exposure, characterized by erythematic and swelling nose with extensive mucous secretion (with or without bleeding). At 6 h post exposure most of the guinea pigs had breathing difficulties, rhonchi and dyspnea and few deaths were noted. These symptoms peaked at 48 h and were noted up to 8 days, associated with few additional deaths. Thereafter, a spontaneous healing was noted, characterized by recovery of respiratory parameters and normal weight gain with almost complete apparent healing within 2 weeks. Histopathological evaluation of lungs and trachea in the surviving GPs at 4 weeks post exposure revealed a dose-dependent residual injury in both lung and trachea expressed by abnormal recovery of the tracheal epithelium concomitant with a dose-dependent increase in cellular volume in the lungs. These abnormal epithelial regeneration and lung remodeling were accompanied with significant changes in protein, LDH, differential cell count and glutathione levels in the bronchoalveolar lavage (BAL). It is suggested that the abnormal epithelial growth and cellular infiltration into the lung as well as the continuous lung inflammation could cause recurrent lung injury similar to that reported for HD exposed human casualties. PMID- 19682478 TI - Peroxynitrite mediates muscle insulin resistance in mice via nitration of IRbeta/IRS-1 and Akt. AB - Accumulating evidence suggests that peroxynitrite (ONOO(-)) is involved in the pathogenesis of insulin resistance. In the current study, we investigated whether insulin resistance in vivo could be mediated by nitration of proteins involved in the early steps of the insulin signal transduction pathway. Exogenous peroxynitrite donated by 3-morpholinosydnonimine hydrochloride (SIN-1) induced in vivo nitration of the insulin receptor beta subunit (IRbeta), insulin receptor substrate (IRS)-1, and protein kinase B/Akt (Akt) in skeletal muscle of mice and dramatically reduced whole-body insulin sensitivity and muscle insulin signaling. Moreover, in high-fat diet (HFD)-fed insulin-resistant mice, we observed enhanced nitration of IRbeta and IRS-1 in skeletal muscle, in parallel with impaired whole body insulin sensitivity and muscle insulin signaling. Reversal of nitration of these proteins by treatment with the peroxynitrite decomposition catalyst FeTPPS yielded an improvement in whole-body insulin sensitivity and muscle insulin signaling in HFD-fed mice. Taken together, these findings provide new mechanistic insights for the involvement of peroxynitrite in the development of insulin resistance and suggest that nitration of proteins involved in the early steps of insulin signal transduction is a novel molecular mechanism of HFD-induced muscle insulin resistance. PMID- 19682480 TI - Activation of Nrf2 by cadmium and its role in protection against cadmium-induced apoptosis in rat kidney cells. AB - Kidney is the primary target organ in chronic cadmium (Cd) toxicity, and oxidative stress plays an important role in this process. The nuclear transcription factor Nrf2 binds to antioxidant response elements (AREs) and regulates genes involved in protecting cells from oxidative damage. Whether kidney cells respond to Cd by activating Nrf2 is unknown. This study was designed to examine the Cd-induced activation of Nrf2 transcriptional activity in a stable rat kidney cell line, NRK-52E, and to investigate the protection this might offer against apoptosis. The cells were treated with 5-20 microM CdCl(2) for 5 h, followed by a recovery period of up to 24 h. A concentration-dependent increase (up to 2.9-fold) in the level of reactive oxygen species was noted upon termination of 5-h Cd treatment. The Nrf2-ARE binding activity also increased and peaked (6.1-fold) at 10 microM Cd concentration. Time-course study revealed that the binding activity increased at 1 h of Cd treatment and peaked 2 h post Cd treatment. Apoptosis was detected 6 h post treatment with Cd and a concentration- and time-dependent increase in the apoptotic cell population occurred during the next 18 h. Over-expression of Nrf2 by transient transfection conferred resistance against Cd-induced apoptosis. Conversely, suppression of Nrf2 expression by specific siRNA resulted in greater sensitivity of the cells to Cd by decreasing the levels of two antioxidant enzymes, hemeoxygenase-1 and glutamate-cysteine ligase. Taken together, these results suggest that in kidney cells the activation of Nrf2 is an adaptive intracellular response to Cd-induced oxidative stress, and that Nrf2 is protective against Cd-induced apoptosis. PMID- 19682479 TI - Arsenic exacerbates atherosclerotic lesion formation and inflammation in ApoE-/- mice. AB - Exposure to arsenic-contaminated water has been shown to be associated with cardiovascular disease, especially atherosclerosis. We examined the effect of arsenic exposure on atherosclerotic lesion formation, lesion composition and nature in ApoE-/- mice. Early post-natal exposure (3-week-old mice exposed to 49 ppm arsenic as NaAsO(2) in drinking water for 7 weeks) increased the atherosclerotic lesion formation by 3- to 5-fold in the aortic valve and the aortic arch, without affecting plasma cholesterol. Exposure to arsenic for 13 weeks (3-week-old mice exposed to 1, 4.9 and 49 ppm arsenic as NaAsO(2) in drinking water) increased the lesion formation and macrophage accumulation in a dose-dependent manner. Temporal studies showed that continuous arsenic exposure significantly exacerbated the lesion formation throughout the aortic tree at 16 and 36 weeks of age. Withdrawal of arsenic for 12 weeks after an initial exposure for 21 weeks (to 3-week-old mice) significantly decreased lesion formation as compared with mice continuously exposed to arsenic. Similarly, adult exposure to 49 ppm arsenic for 24 weeks, starting at 12 weeks of age increased lesion formation by 2- to 3.6-fold in the aortic valve, the aortic arch and the abdominal aorta. Lesions of arsenic-exposed mice displayed a 1.8-fold increase in macrophage accumulation whereas smooth muscle cell and T-lymphocyte contents were not changed. Expression of pro-inflammatory chemokine MCP-1 and cytokine IL-6 and markers of oxidative stress, protein-HNE and protein-MDA adducts were markedly increased in lesions of arsenic-exposed mice. Plasma concentrations of MCP-1, IL 6 and MDA were also significantly elevated in arsenic-exposed mice. These data suggest that arsenic exposure increases oxidative stress, inflammation and atherosclerotic lesion formation. PMID- 19682481 TI - Ciguatoxin-induced catecholamine secretion in bovine chromaffin cells: mechanism of action and reversible inhibition by brevenal. AB - Ciguatoxin (P-CTX-1B) from the dinoflagellate Gambierdiscus toxicus, belongs to the family of polyether neurotoxins responsible for the neurological poisoning disorder ciguatera. Although it is the most widespread marine-borne disease affecting humans, there is no current FDA-approved treatment available except for symptomatic therapies. In this paper, we report that P-CTX-1B promotes catecholamine secretion from bovine chromaffin cells, an effect that is insensitive to concomitant activation of capacitative Ca(2+) entry. Moreover, we confirm that brevenal, a polyether from the dinoflagellate Karenia brevis, blocks P-CTX-1B-induced catecholamine secretion. This effect is partially reversible. Our results therefore raise the prospect of finding functional antagonists for P CTX-1B that could be useful for the treatment of ciguatera. PMID- 19682482 TI - Characteristic profiles of Ciguatera toxins in different strains of Gambierdiscus spp. AB - Ciguatera fish poisoning characterizes the intoxication caused by consumption of fish from tropical and subtropical areas, which have accumulated ciguatoxins (CTXs). The observed pattern of ciguatoxins in fish highly depends on the marine region and the causative organisms. It is evident that differences exist between ciguatoxins produced by certain strains of the dinoflagellate Gambierdiscus toxicus and other Gambierdiscus spp. In this context cultured strains purchased from the Provasoli-Guillard National Center for Culture of Marine Phytoplankton (CCMP) and strains from Vietnam were analyzed. Besides, lyophilized samples of several Gambierdiscus spp. from the National Oceanic and Atmospheric Administration (NOAA), USA and lyophilized samples of G. toxicus from Vietnam were analyzed. The latter has been cultured at different salinities. We observed differences between the toxin ratios of the analogues in the strain from Vietnam depending on the salinity. The CTX profiles of the Vietnamese samples were compared with cultures of Gambierdiscus spp. from CCMP and the National Oceanic and Atmospheric Administration (NOAA) resulting in an overview of toxins in cultures from different regions. Hence, it was obvious that the strain from Vietnam forms a characteristic CTX profile which is not directly comparable to CTX pattern observed in other tropical marine regions. PMID- 19682483 TI - Maturation-associated changes in toxicity of the pufferfish Takifugu poecilonotus. AB - From October 2006 to December 2007, wild specimens of the pufferfish Takifugu poecilonotus (93 females, 45 males) were collected from the Ariake Sea. Tissue toxicity was examined by mouse bioassay, and tetrodotoxin (TTX) content in the blood plasma by enzyme-linked immunosorbent assay. The relationship between toxicity and maturation was investigated based on changes in the gonadosomatic index: December-March in females and November-March in males, the 'maturation period'; April, 'just after spawning'; and the other months, the 'ordinary period'. Toxicity of both sexes was high throughout the year, but sharply declined in April. In all tissues examined (skin, liver, and ovary) other than testis, toxicity exceeded 1000 MU/g or 10,000 MU/individual in many individuals. Seasonal profiles of tissue toxicity differed markedly between sexes. In females, liver toxicity was high during the ordinary period, and ovary toxicity was high during the maturation period. In males, little maturation-associated change in the toxin distribution was observed. Plasma TTX levels were similar between the sexes (1.59-15.1 MU/ml), and fluctuated largely throughout the year without corresponding changes in tissue toxicity. The percentage of TTX binding to high molecular-weight substances in the plasma varied in association with maturation; the binding ratio fluctuated at relatively low levels during the ordinary period, and stabilized at a high level during the maturation period. PMID- 19682484 TI - Neurotoxic and cytotoxic effects of venom from different populations of the Egyptian Scorpio maurus palmatus. AB - Neurotoxic and cytotoxic effects of venoms from Scorpio maurus palmatus taken from different populations were assessed for geographic based variability in toxicity, and to evaluate their insecticidal potency. Scorpions were collected from four regions. Three locations were mutually isolated pockets in the arid area of Southern Sinai. The fourth sample was collected from a population inhabiting the semi-arid environment of Western Mediterranean Coastal Desert. The neurotoxic (paralytic) effect of the venom from each population was assayed by its ability to induce permanent disability in adult cockroaches within 3h. Venom was applied using microinjection techniques through an intersegmental membrane. Probit analysis was used to calculate the Paralytic Effective Dose (PED(50), ng/100mg). Levels of glutathione, lipid peroxidation, protein carbonyl content and nitric oxide, as well as the activities of superoxide dismutase, catalase and cholinesterase, were measured to assess the cytotoxicity of the venom. The results show that the injected venom from each population induced obvious spasticity, followed by flaccid paralysis. All the tested biochemical parameters, except glutathione content, revealed significant differences in toxicity in venom taken from the different scorpion populations. We conclude that (i) the venom of this scorpion has significant neurotoxic and cytotoxic effects on insect cells, (ii) its efficacy, as assessed by the PED(50) unit, exhibited variation across its geographic range, and (iii) components in the venom may have the potential for being developed into effective and environmentally friendly bioinsecticides. PMID- 19682485 TI - Vision in autism spectrum disorders. AB - Autism spectrum disorders (ASDs) are developmental disorders which are thought primarily to affect social functioning. However, there is now a growing body of evidence that unusual sensory processing is at least a concomitant and possibly the cause of many of the behavioural signs and symptoms of ASD. A comprehensive and critical review of the phenomenological, empirical, neuroscientific and theoretical literature pertaining to visual processing in ASD is presented, along with a brief justification of a new theory which may help to explain some of the data, and link it with other current hypotheses about the genetic and neural aetiologies of this enigmatic condition. PMID- 19682486 TI - Curvature coding in illusory contours. AB - We have employed the shape frequency and shape-amplitude after-effects (SFAE and SAAE) to investigate: (i) whether the shapes of illusory and real curves are processed by the same or different mechanisms, and (ii) the carrier-tuning properties of illusory curvature mechanisms. The SFAE and SAAE are the phenomena in which adaptation to a sinusoidal-shaped contour results in a shift in, respectively, the perceived shape-frequency and perceived shape-amplitude of a test contour in a direction away from that of the adapting stimulus. Both after effects are believed to be mediated by mechanisms sensitive to curvature (Gheorghiu & Kingdom, 2007a, 2009; see also Hancock & Peirce, 2008). We observed both shape after-effects in sinusoidally-shaped illusory contours defined by phase-shifted line-grating carriers. We tested whether illusory and real contours were mediated by the same or different mechanisms by comparing same adaptor-and test with different adaptor-and-test combinations of real and illusory contours. Real contour adaptors produced after-effects in illusory contour tests that were as great as, or even greater than those produced by illusory contour adaptors. However, illusory contour adaptors produced much weaker after-effects in real contour tests than did real contour adaptors. This asymmetry suggests that illusory contour curves are encoded by a sub-set of mechanisms sensitive to real contour curves. We also examined the carrier-tuning properties of illusory contour curvature processing using adaptor and test illusory contours that differed in the luminance contrast-polarity, luminance scale and orientation of the carriers. We found no selectivity to any of these dimensions for either even symmetric or odd-symmetric line-gratings carriers, even though selectivity to these dimensions was found for real contours. PMID- 19682487 TI - Contribution of configural information in a direction discrimination task: evidence using a novel masking paradigm. AB - Understanding how structure and motion information contribute to the perception of biological motion is often studied with masking techniques. Current techniques in masking point-light walkers typically rely on adding surrounding masking dots or altering phase relations between joints. Here, we demonstrate the use of novel stimuli that make it possible to determine the noise level at which the local motion cues mask the opposing configural cues without changing the number of overall points in the display. Results show improved direction discrimination when configural cues are present compared to when the identical local motion signals are present but lack configural information. PMID- 19682489 TI - Cellular senescence of human mammary epithelial cells (HMEC) is associated with an altered MMP-7/HB-EGF signaling and increased formation of elastin-like structures. AB - The extracellular matrix (ECM) and a complex interplay of cell-to-cell and cell to-matrix (ECM) interactions provide important platforms to determine cellular senescence and a potentially tumorigenic transformation of normal human mammary epithelial cells (HMEC). An enhanced formation of extracellular filaments, consisting of elastin-like structures, in senescent post-selection HMEC populations was paralleled by a significantly increased expression of its precursor protein tropoelastin and matched with a markedly elevated activity of the cross-linking enzyme family of lysyl oxidases (LOX). RNAi experiments revealed both the ECM metalloproteinase MMP-7 and the growth factor HB-EGF as potential effectors of an increased tropoelastin expression. Moreover, co localization of MMP-7 and HB-EGF as well as a concomittant downstream signaling via Fra-1 indicated a possible association between the reduced MMP-7 enzyme activity and an impaired HB-EGF processing, resulting in an enhanced tropoelastin synthesis during senescence of HMEC. In agreement with previous work, these findings suggested an important influence of the extracellular proteinase MMP-7 on the aging process of HMEC, affecting both extracellular remodeling as well as intracellular signaling pathways. PMID- 19682488 TI - Defective responses to oxidative stress in protein l-isoaspartyl repair-deficient Caenorhabditis elegans. AB - We have shown that Caenorhabditis elegans lacking the PCM-1 protein repair l isoaspartyl methyltransferase are more sensitive to oxidative stress than wild type nematodes. Exposure to the redox-cycling quinone juglone upon exit from dauer diapause results in defective egg-laying (Egl phenotype) in the pcm-1 mutants only. Treatment with paraquat, a redox-cycling dipyridyl, causes a more severe developmental delay at the second larval stage in pcm-1 mutants than in wild-type nematodes. Finally, exposure to homocysteine and homocysteine thiolactone, molecules that can induce oxidative stress via distinct mechanisms, results in a more pronounced delay in development at the first larval stage in pcm-1 mutants than in wild-type animals. Homocysteine treatment also induced the Egl phenotype in mutant but not wild-type nematodes. All of the effects of these agents were reversed upon addition of vitamin C, indicating that the developmental delay and egg-laying defects result from oxidative stress. Furthermore, we have demonstrated that a mutation in the gene encoding the insulin-like receptor DAF-2 suppresses the Egl phenotype in pcm-1 mutants treated with juglone. Our results support a role of PCM-1 in the cellular responses mediated by the DAF-2 insulin-like signaling pathway in C. elegans for optimal protection against oxidative stress. PMID- 19682490 TI - Computer-delivered, parent-involvement intervention to prevent substance use among adolescent girls. AB - OBJECTIVE: To test a computer-delivered program for preventing substance use among adolescent girls. METHODS: Randomly, 916 girls aged 12.76+/-1.0 years and their mothers were assigned to an intervention arm or to a test-only control arm. Intervention-arm dyads engaged in exercises to improve the mother-daughter relationship, build girls' substance use prevention skills, and reduce associated risk factors. Study outcomes were girls' and mothers' substance use and mediator variables related to girls' substance use risk and protective factors. The study was conducted between September 2006 and February 2009 with participants from greater New York City, including southern Connecticut and eastern New Jersey. RESULTS: At 2-year follow-up and relative to control-arm girls, intervention-arm girls reported lower relevant risk factors and higher protective factors as well as less past 30-day use of alcohol (p<0.006), marijuana (p<0.016), illicit prescription drugs (p<0.03), and inhalants (p<0.024). Intervention-arm mothers showed more positive 2-year outcomes than control-arm mothers on variables linked with reduced risks of substance use among their daughters, and mothers reported lower rates of weekly alcohol consumption (p<0.0001). CONCLUSIONS: A computer delivered prevention program for adolescent girls and their mothers was effective in changing girls' risk and protective factors and girls' and mothers' substance use behavior. PMID- 19682491 TI - Late embryonic exposure to all-trans retinoic acid induces a pattern of motor deficits unrelated to the developmental stage. AB - The present study extends previous investigations examining the behavioral outcomes of all-trans retinoic acid (RA) exposure at embryonic (E) days 14-16. A sublethal dose (2.5 mg/kg b.w.) compatible with high neonatal survival sufficient to supply offspring for later behavioral testing, was used. The results show that E14-16 RA exposure, similar to E8-10 or E11-13 (previous studies), impairs locomotor activity (open field test) as well as motor coordination and motor learning (rotarod/accelerod task) in young-adult rats. The results provide further evidence that RA exposure induces a pattern of motor deficits which are not strictly related to the embryonic stage, compatible with the protracted developmental profile of the cerebellum. PMID- 19682493 TI - A simple method for quantitative calcium imaging in unperturbed developing neurons. AB - Calcium imaging has been widely used to address questions of neuronal function and development. To gain deeper insights into the actions of calcium as a second messenger, but also to measure synaptic function, it is necessary to quantify the level of calcium at rest and during calcium transients. While quantification of calcium levels is straightforward when using ratiometric calcium indicators, these dyes have several draw-backs due to their short wavelength excitation spectra, such as light scattering and cytotoxicity. In contrast, many single wavelength indicators exhibit superior photostability, low phototoxicity, extended dynamic ranges and very high signal to noise ratios. However, quantifying calcium levels in unperturbed neurons has not been performed with these indicators. Here, we explore a new approach for determining the calcium concentration at rest as well as calcium rises during evoked and spontaneous neuronal activity in unperturbed developing neurons using a single-wavelength calcium indicator. We show that measuring the maximal fluorescence at the end of an imaging experiment allows determining calcium levels with high resolution. Specifically, we assessed the limits of calcium measurements with a CCD camera in small neuronal processes and found that even in small diameter dendrites and spines the intracellular calcium concentration and its changes can be estimated accurately. This approach may not only allow mapping patterns of neuronal activity quantitatively with the resolution of single synapses and a few tens of milliseconds, but also facilitate investigating the role of calcium as a second messenger. PMID- 19682492 TI - Real-time artifact filtering in continuous VEPs/fMRI recording. AB - Continuous recording of Visual Evoked Potentials (VEPs) and functional Magnetic Resonance Imaging (fMRI) exploits the VEPs high temporal resolution and the fMRI high spatial resolution. In this work, we present a new method of continuous VEPs/fMRI recording to study visual function in seven normal subjects. Our real time artifact filtering is characterized by a procedure based on an analytical study of echo-planar imaging (EPI) sequence parameters related electro encephalogram (EEG)-artifact shapes. The magnetic field artifacts were minimized by using a dedicated amagnetic device and by a subtraction algorithm that takes into account the EPI sequence parameters. No significant decrease in signal-to noise ratio was observed in case of EEG recording simultaneously with MR acquisition; similarly, transient and steady-state VEPs parameters were comparable during fMRI acquisition and in the off-phase of fMRI recording. We also applied this method to one patient with optic neuritis, and, compared with controls, found different results. We suggest that our technique can be reliably used to investigate the function of human visual cortex and properly correlate the electrophysiological and functional neuroimaging related changes. PMID- 19682494 TI - PKCtheta and Itk functionally interact during primary mouse CD3+ T cell activation. AB - PKCtheta serine/threonine and Itk tyrosine protein kinases have been implicated in T lymphocyte signal transmission. We observed a PKCtheta/Itk complex after T cell activation, raising the possibility that PKCtheta and Itk might interact functionally during T cell development and response. To address this question PKCtheta/Itk double knockout mice were generated and T cell activation responses were compared to single deficiencies as well as to wild type controls. Consistent with previous reports, Itk and PKCtheta are required in modulating CD3(+) T cell cytokine secretion responses ex vivo. Itk- and PKCtheta-deficient cells show impaired NFAT/AP-1 and NF-kappaB transactivation responses, however the combined loss, did not exceed but partially rescue the strong NFAT and NF-kappaB activation defects observed in Itk(-/-) single-deficient T cells. Taken together, this provides evidence for a more complex functional crosstalk between Itk and PKCtheta during T cell receptor signalling then previously anticipated. PMID- 19682495 TI - An in vivo model of priming of antigen-specific human CTL by Mo-DC in NOD/Shi scid IL2rgamma(null) (NOG) mice. AB - In vivo assay to evaluate anti-cancer immunotherapy at the pre-clinical phase is eagerly needed. We currently established xenotransplantation-based method to analyze in vivo priming of cancer-antigen-specific human cytotoxic T lymphocytes (CTLs). We transplanted human peripheral T cells and analyzed priming of CTLs in NOG mice. Half of the mice engrafted with bulk lymphocytes including CD4(+) T cells died before analysis probably due to xenoreactive graft versus host disease. All of the mice engrafted with purified CD8(+) T cells survived until the analysis, and successful engraftment was observed in 80% of recipient mice. Thus, transfer of purified CD8(+) T cells is sufficient and safer than that of bulk lymphocytes. To add antigenic stimulation to the CD8(+) T cells in vivo, injection of antigenic peptide-loaded and monocyte-derived autologous dendritic cells (DCs) was simultaneously done and repeated 7 days later. The DC-based vaccinization resulted in efficient priming of HLA class I-restricted and MART1, WT1 or CMV peptides-specific CTLs in the recipient mice. This system may be useful to evaluate the stimulation of antigen-specific human CTLs in vivo. PMID- 19682496 TI - Modulation of inflammation through IL-17 production by gammadelta T cells: mandatory in the mouse, dispensable in humans? AB - Recent studies suggest that gammadelta T cells are innate IL-17 producers owing to unique features of their developmental program. A key contribution of this subset to T helper 17 responses has been also suggested by numerous physiopathological studies mainly performed in mouse models. In the present review, we will summarize the main features of IL-17-producing gammadelta T cells and highlight the similarities and differences between murine gammadelta T cells and their human counterparts. PMID- 19682497 TI - Altered apoptosis of inflammatory neutrophils in MMP-9-deficient mice is due to lower expression and activity of caspase-3. AB - Matrix metalloproteinase 9 (MMP-9) is a Zn(2+)-dependent endopeptidase that degrades some of the components of basement membranes and extracellular matrix and thus participates in leukocyte infiltration during inflammation. In a model of zymosan peritonitis, neutrophil infiltration in MMP-deficient (MMP-9(-/-)) mice was significantly weaker at the time of their maximal influx in wild-type mice (6h). However, during the late stages of peritonitis (24h) an extended accumulation of neutrophils was observed in MMP-9(-/-)versus the wild-type mice. Recently, we reported that the ratio of apoptosis of inflammatory leukocytes is impaired in MMP-9(-/-) mice during late peritonitis and the process depends on COX-1-driven PGE(2). Here we scrutinized the alterations in apoptotic mechanisms by comparisons between MMP-9(-/-) and the wild-type mice. Altered apoptosis occurred only during late (24h) peritonitis and concerned only neutrophils, and not macrophages, mast cells or lymphocytes. Furthermore, expression and activity of caspases was altered in MMP-9(-/-) animals, delayed for caspase-8 and -9, and decreased in the case of caspase-3. Also the expression of Bax/Bcl-2 proteins was changed in MMP-9(-/-) mice. These changes, and in particular the impaired neutrophil apoptosis and weaker caspase-3 activity, were restored by the selective COX-1 inhibition. We conclude that in mice lacking MMP-9 the enhanced COX-1-PGE(2) decreases caspase-3 expression and activity leading to impaired apoptosis of inflammatory neutrophils resulting in abnormal accumulation of the cells at the inflammatory focus. The data also reinforce the notion that MMP-9 is a key enzyme in neutrophil biology. PMID- 19682498 TI - A SYBR Green I real-time RT-PCR assay for detection and differentiation of influenza A(H1N1) virus in swine populations. AB - The novel influenza A(H1N1) virus that emerged recently in Mexico has spread rapidly to many countries and initiated a human pandemic. It would be interesting to determine whether the virus has existed in, or will spread to, the swine population. However, it is difficult to differentiate the virus from some swine influenza viruses. In this study, a SYBR Green I real-time RT-PCR assay was designed for detection and differentiation of influenza A(H1N1) virus from some swine influenza viruses, by comparing the amplification of two pairs of primers corresponding to influenza A(H1N1) virus and some swine influenza viruses, respectively. The assay was evaluated using online analysis, identified influenza viruses and clinical samples. The results indicated that the assay has high sensitivity and specificity to detect influenza A(H1N1) virus, and is able to differentiate it from some swine influenza viruses. This, in turn, could provide essential epidemiological information for risk analysis and decision making in combating the disease, and stimulate research to differentiate pathogens similar to each other using the same method. PMID- 19682501 TI - Early life programming of fear conditioning and extinction in adult male rats. AB - The early rearing environment programs corticolimbic function and neuroendocrine stress reactivity in adulthood. Although early environmental programming of innate fear has been previously examined, its impact on fear learning and memory later in life remains poorly understood. Here we examined the role of the early rearing environment in programming fear conditioning and extinction in adult male rats. Pups were subjected to maternal separation (MS; 360 min), brief handling (H; 15 min), or animal facility rearing (AFR) on post-natal days 2-14. As adults, animals were tested in a 3-day fear learning and memory paradigm which assessed the acquisition, expression and extinction of fear conditioning to an auditory cue; the recall of extinction was also assessed. In addition, contextual fear was assessed prior to cued extinction and its recall. We found that the acquisition of fear conditioning to the cue was modestly impaired by MS. However, no early rearing group differences were observed in cue-induced fear expression. In contrast, both the rate of extinction and extinction recall were attenuated by H. Finally, although contextual fear was reduced after extinction to the cue, no differences in context-induced fear were observed between the early rearing groups. These results add to a growing body of evidence supporting an important role for early environmental programming of fear conditioning and extinction. They also indicate that different early rearing conditions can program varying effects on distinct fear learning and memory processes in adulthood. PMID- 19682502 TI - Transcriptional memory and switching in the Plasmodium falciparumrif gene family. AB - The human malaria parasite Plasmodium falciparum expresses erythrocyte-surface directed variant antigens which are important virulence factors. Many are transcribed from multigene families and presumably their mode of expression is strictly controlled to guarantee immune evasion in the human host. In order to elucidate the dynamics of rif transcription and to investigate if rif switching is comparable to var switching we monitored rif variant gene expression in parasites with different cytoadhesive properties as well as after a number of reinvasions. We found identical transcripts in parasite lines with different adhesive phenotypes suggesting that rif genes do not have a critical role in determining the cytoadhesion specificity of infected erythrocytes. We show for the first time that rif genes may show a conserved mode of transcription, maintaining the previously dominant rif transcript in subsequent reinvasions, but also observed rapid switching at rates up to 45% per generation, much higher than for the var gene family. PMID- 19682500 TI - Reversal of chlorpyrifos neurobehavioral teratogenicity in mice by nicotine administration and neural stem cell transplantation. AB - Identifying the mechanisms underlying the adverse effects of developmental neurotoxicants enables the design of therapies that can potentially reverse neurobehavioral deficits in adulthood. We administered chlorpyrifos (CPF), a model organophosphate pesticide to pregnant mice and identified visuospatial deficits in adult offspring using performance in the Morris maze. We then evaluated two strategies to reverse the effects, nicotine administration and transplantation of neural stem cells. Daily administration of nicotine prior to behavioral testing did not alter maze performance by itself, but completely reversed the deficits evoked by prenatal CPF exposure. Similarly, control animals grafted with neural stem cells in adolescence did not show any alterations in behavioral performance as adults, but the grafts completely reversed the effects of prenatal CPF treatment. This study thus provides a model for the development and application of both pharmacologic and cell-based therapies to offset the effects of neurobehavioral teratogens. PMID- 19682503 TI - Growth hormone-releasing peptide-2 stimulates secretion and synthesis of adrenocorticotropic hormone in mouse pituitary. AB - Growth hormone (GH)-releasing peptides (GHRPs) are synthetic peptides which induce strong GH release in both animals and humans. Among them, GHRP-2 is known to stimulate GH release by acting at both hypothalamic and pituitary sites, but also induces adrenocorticotropic hormone (ACTH) release in healthy subjects. GHRP 2 may stimulate ACTH release directly via GHRP receptor type 1a in ACTH-producing tumors. GHRP-2 increases ACTH secretion in rat in vivo, but not ACTH release from rat primary pituitary cells. In the present study, in order to elucidate the mechanism underlying ACTH secretion by GHRPs, mouse pituitary cells were stimulated by GHRP-2. GHRP receptor mRNA was expressed in the mouse pituitary, and GHRP-2 directly stimulated secretion and synthesis of ACTH in the mouse anterior pituitary cells. GHRP-2 increased intracellular cyclic AMP production. H89, a potent protein kinase A (PKA) inhibitor, and bisindolylmaleimide I, a selective protein kinase C (PKC) inhibitor, inhibited the GHRP-2-induced ACTH release, and that H89, but not bisindolylmaleimide I, inhibited the GHRP-2 induced proopiomelanocortin mRNA levels. Together, the GHRP-2-induced ACTH release was regulated via both PKA and PKC pathways in the mouse pituitary cells, while ACTH was synthesized by GHRP-2 only via the PKA pathway. PMID- 19682499 TI - Reverse genetics technology for Rift Valley fever virus: current and future applications for the development of therapeutics and vaccines. AB - The advent of reverse genetics technology has revolutionized the study of RNA viruses, making it possible to manipulate their genomes and evaluate the effects of these changes on their biology and pathogenesis. The fundamental insights gleaned from reverse genetics-based studies over the last several years provide a new momentum for the development of designed therapies for the control and prevention of these viral pathogens. This review summarizes the successes and stumbling blocks in the development of reverse genetics technologies for Rift Valley fever virus and their application to the further dissection of its pathogenesis and the design of new therapeutics and safe and effective vaccines. PMID- 19682504 TI - Hsp90 transcriptionally and post-translationally regulates the expression of NDRG1 and maintains the stability of its modifying kinase GSK3beta. AB - N-myc downstream-regulated gene 1 (NRDG1) is a stress-induced protein whose putative function is suppression of tumor metastasis. A recent proteonomic study showed NDRG1 interacts with the molecular chaperone heat shock protein 90 (Hsp90). From their reported association, we investigated if NDRG1 is dependent on Hsp90 for its stability and is therefore a yet unidentified Hsp90 client protein. Here, we demonstrate that endogenous NDRG1 and Hsp90 physically associate in hepatocellular cancer cell lines. However, geldanamycin (GA) mediated inhibition of Hsp90 did not disrupt their interaction or result in NDRG1 protein destabilization. On the contrary, inhibition of Hsp90 led to a transcriptional increase of NDRG1 protein which was associated with cell growth arrest. We also observed that GA inhibited the phosphorylation of NDRG1 by targeting its regulating kinases, serum- and glucocorticoid-induced kinase 1 (SGK1) and glycogen synthase kinase 3 beta (GSK3beta). We demonstrate that in the presence of GA, GSK3beta protein and activity were decreased thus indicating that Hsp90 is necessary for GSK3beta stability. Taken together, our data demonstrate that NDRG1 is not a classic client protein but interacts with Hsp90 and is still dually regulated by Hsp90 at a transcriptional and post-translational level. Finally, we suggest for the first time GSK3beta as a new client protein of Hsp90. PMID- 19682505 TI - Selective adsorption of hydrocarbon-oxidizing Rhodococcus cells in a column with hydrophobized poly(acrylamide) cryogel. AB - A method for selective adsorption of Rhodococcus cells in the column with hydrophobized poly(acrylamide) cryogel (cryoPAAG) was developed that allowed rhodococci separation from mixed bacterial populations and their effective concentration within a sponge-like gel matrix. Hydrophobization of cryoPAAG using the n-dodecane graft (C12) was performed to enhance the adhesion of Rhodococcus cells to the cryogel; this was suggested by our finding that alkanotrophic rhodococci possess high adhesive activity (91-98%) towards n-alkanes, whereas other Gram-positive and Gram-negative bacteria tested did not adhere strongly to hydrocarbons. The selective index of the hydrophobic C12-cryoPAAG column for Rhodococcus cells was 72% that ensured their separation from complex bacterial cultures. Respirometry results using the Columbus Micro-Oxymax respirometer showed that the maximal respiratory activity of C12-cryoPAAG-immobilized Rhodococcus cells incubated with petroleum hydrocarbons was 1.6-1.8 times higher than that of freely suspended cells, and this correlated with the largest immobilized cell number. Moreover, high respiration rates were maintained over 3 weeks of incubation, indicating a considerable functional stability of the cryoPAAG-immobilized biocatalyst developed. PMID- 19682506 TI - Social ties and cardiovascular function: an examination of relationship positivity and negativity during stress. AB - The quality and quantity of one's relationships have been reliably linked to morbidity and mortality. More recently, studies have focused on links between relationships and cardiovascular reactivity as a physiological mechanism via the stress-buffering hypothesis. However, not all social relationships are consistently positive which points to the importance of a more comprehensive examination of relationship that includes negative qualities. In this study, we manipulated relationship positivity and negativity with an experimenter and examined its influence on cardiovascular reactivity. Results revealed that relationship positivity was associated with lower systolic blood pressure (SBP) reactivity for men and women. Relationship negativity, on the other hand, was related to less of an increase in diastolic blood pressure (DBP) reactivity in men. Internal analyses showed that perceptions of positivity and negativity interacted such that high positivity/high negativity perceptions (ambivalence) were related to the highest SBP reactivity. Results of this study suggest that the quality of one's relationships is an important moderator of cardiovascular reactivity during stress. PMID- 19682508 TI - Cerebral heme oxygenase 1 and 2 spatial distribution is modulated following injury from hypoxia-ischemia and middle cerebral artery occlusion in rats. AB - The regional and cellular distribution of heme oxygenase (HO)-1 and -2 following cerebral ischemia has not been ascertained. Employing the transient middle cerebral artery occlusion (MCAO) and hypoxia-ischemia (HI) models of unilateral brain injury, the aim was to elucidate immunolocalization of HO-1 and HO-2. Animals were sacrificed 3 days post-ischemia and immunohistochemistry and Western blotting were utilized to determine HO-1 and HO-2 expression. In the ipsilateral hemisphere following HI, HO-1 immunoreactivity was significantly upregulated in many neuronal and glial populations (including the cortex, hippocampus and thalamus). HO-1 was also detected in macrophages/microglia within the infarct. In addition to widespread neuronal HO-2 labelling, HO-2 was also expressed in vascular endothelial cells. Inflammatory cells within the infarct of MCAO and HI animals were surprisingly immunoreactive for HO-2, but only HI animals had significantly elevated HO-2 protein expression in the ipsilateral hemisphere. This may be due to the presence of global hypoxia in the HI model which can upregulate vascular endothelial growth factor and subsequent proliferation of endothelial cells. This report of HO-2 protein expression upregulation following HI coupled with an increase in HO-1 immunoreactivity suggests that this response may be implicated in reducing cell death or repairing damage induced by cerebral ischemia. PMID- 19682507 TI - Stereo Navi 2.0: software for stereotaxic surgery of the common marmoset (Callithrix jacchus). AB - Recently, we reported our web-accessible digital brain atlas of the common marmoset (Callithrix jacchus) at http://marmoset-brain.org:2008. Using digital images obtained during construction of this website, we developed stand-alone software for navigation of electrodes or injection needles for stereotaxic electrophysiological or anatomical experiments in vivo. This software enables us to draw lines on exchangeable section images, measure the length and angle of lines, superimpose a stereotaxic reference grid on the image, and send the image to the system clipboard. The software, Stereo Navi 2.0, is freely available at our brain atlas website. PMID- 19682509 TI - Cellular distribution of the nicotinic acetylcholine receptor alpha7 subunit in rat hippocampus. AB - The hippocampus is a region of the mammalian brain that has been extensively studied due to its role in many forms of memory. To better understand hippocampal function, significant attention has focused upon the cellular distribution of ligand-gated ion channels. Despite strong cholinergic innervation from the basal forebrain and a dense expression of nicotinic acetylchoine receptors (nAChRs), the cellular distribution of subunits forming these receptors has received little attention. We used organotypic hippocampal slice cultures (OHSCs) to study native alpha7 subunits, which, unlike other nAChR subunits, form a homomeric receptor. Cell-surface biotinylation, cross-linking of surface proteins, and sub-cellular fractionation all revealed a very limited presence of the subunit at the plasma membrane. In contrast, subunits of other receptors displayed significant surface expression. Notably, subunits in adult hippocampal tissue were distributed in a fashion similar to that observed in OHSCs. To monitor alpha7 subunits contained in functional nAChRs, a colourimetric assay using alpha-bungarotoxin (a specific alpha7 nAChR antagonist) was developed, and revealed a majority of binding at the cell surface. To change alpha7 subunit distribution, OHSCs were treated with compounds known to affect other ionotropic receptors-insulin, genistein, and elevated external K(+); however, neither subunit surface expression nor antagonist binding was affected. Our data reveal that hippocampal neurons possess a large internal population of alpha7 subunits under basal conditions, which persists during stimuli affecting tyrosine phosphorylation or neuronal activity. The nature of the internal pool of alpha7 subunits remains to be determined, but should have important implications for hippocampal activity. PMID- 19682510 TI - Characterization of the nuclear and nucleolar localization signals of bovine herpesvirus-1 infected cell protein 27. AB - Bovine herpesvirus-1 infected cell protein 27 (BICP27) was detected predominantly in the nucleolus. The open reading frame of BICP27 was fused with the enhanced yellow fluorescent protein (EYFP) gene to investigate its subcellular localization in live cells and BICP27 was able to direct monomeric, dimeric or trimeric EYFP exclusively to the nucleolus. By constructing a series of deletion mutants, the putative nuclear localization signal (NLS) and nucleolar localization signal (NoLS) were mapped to (81)RRAR(84) and (86)RPRRPRRRPRRR(97) respectively. Specific deletion of the putative NLS, NoLS or both abrogated nuclear localization, nucleolar localization or both respectively. Furthermore, NLS was able to direct trimeric EYFP predominantly to the nucleus but excluded from the nucleolus, whereas NoLS targeted trimeric EYFP primarily to the nucleus, and enriched in the nucleolus with faint staining in the cytoplasm. NLS+NoLS directed trimeric EYFP predominantly to the nucleolus with faint staining in the nucleus. Moreover, deletion of NLS+NoLS abolished the transactivating activity of BICP27 on gC promoter, whereas deletion of either NLS or NoLS did not. The study demonstrated that BICP27 is a nucleolar protein, adding BICP27 to the growing list of transactivators which localize to the nucleolus. PMID- 19682511 TI - Involvement of mitochondrial permeability transition in hepatitis B virus replication. AB - The HBx protein of human hepatitis B virus (HBV) activates a calcium-dependent kinase pathway which is essential for the viral replication. In this study, we found that HBx expression in the absence of other HBV proteins and in the context of HBV replication decreased the mitochondrial calcein-AM/CoCl(2) signals by 10% and 14% in HepG2 cells and by 15% and 10% in Huh7 cells, respectively. This indicates that HBx can induce mitochondrial permeability transition (MPT) and cause calcium effusion into the plasma. In addition, RNA interference of Cylophilin D decreased HBx-induced MPT and suppressed HBV DNA replication by 41% in HepG2 cells. Our results suggest that HBx expression can induce MPT and facilitate HBV DNA replication. PMID- 19682512 TI - Clinical experience with anthracycline antibiotics-HPMA copolymer-human immunoglobulin conjugates. AB - This paper reviews an early clinical experience with anthracycline (epirubicin; Epi or doxorubicin; Dox) containing an N-(2-hydroyxypropyl)methacrylamide copolymer carrier targeted with autologous or commercial human immunoglobulin in six patients aged 28-55 suffering from therapy-resistant metastatic cancer. More than 100 biochemical, hematological and immunological parameters, including nine tumor markers, were tested in blood samples taken 24 h after the first and up to 10 months after the last application. The intravenous application proceeded without serious adverse or side effects and did not require hospitalization. Cardiotoxicity was not observed. Four of six monitored patients attained stabilization of disease (liver ultrasound scan and bone computer tomography) with a very good quality of life lasting from seven up to 18 months. Positive response to the treatment was, among others, evaluated as decreased CA 15-3 and CEA tumor markers. In three of five tested patients the serum level of C-reactive protein was temporarily increased 72 h after the treatment. A stable or elevated number of peripheral blood reticulocytes together with activation of natural killer (NK) cells and lymphokine-activated killer (LAK) cells supports the data previously obtained in experimental animals pointing to a dual role, i.e. the cytotoxic and immunomobilizing character of doxorubicin-HPMA conjugates. PMID- 19682513 TI - Polymeric carriers: preclinical safety and the regulatory implications for design and development of polymer therapeutics. AB - Since the early 1990s polymer-protein conjugates (included PEGylated enzymes and cytokines), polymeric drugs and polymeric sequestrants have been entering the market as innovative polymer-based therapeutics. Initially these products were most frequently developed as novel anticancer agents; indeed they can be considered first generation "nanomedicines". More recently, a much broader range of life-threatening and debilitating diseases (e.g. viral infections, arthritis, multiple sclerosis and hormone abnormalities) have been targeted via intravenous (i.v.), subcutaneous (s.c.) or oral routes of administration. Given the increasing novelty of polymeric materials proposed for development as second generation polymer therapeutics (with increasing complexity of conjugate composition), and the growing debate as to the safety of nanomedicines per se, the need for evolution of an appropriate regulatory framework is at the forefront of the scientific discussion. The adequacy of the current tests and models used to define safety are also constantly being reviewed. Here we describe the current status and future challenges in relation to these issues. PMID- 19682514 TI - Design and development of IT-101, a cyclodextrin-containing polymer conjugate of camptothecin. AB - IT-101 (Insert Therapeutics-101) is a linear, cyclodextrin-containing polymer conjugate of camptothecin (CPT). When formulated properly, the polymer conjugate self-assembles into nanoparticles of ca. 30 nm diameter and near neutral zeta potential. The nanoparticles show long circulation half-lives in animals and humans and localize in tumors. The nanoparticles enter the tumor cells and slowly release the CPT causing them to disassemble into individual polymer chains that are sufficiently small to be cleared renally. IT-101 is currently being investigated in human clinical trials. Here, the design and development of IT-101 is described with emphasis on features distinguishing it from other polymer containing therapeutics. PMID- 19682515 TI - Functional polymers as therapeutic agents: concept to market place. AB - Biologically active synthetic polymers have received considerable scientific interest and attention in recent years for their potential as promising novel therapeutic agents to treat human diseases. Although a significant amount of research has been carried out involving polymer-linked drugs as targeted and sustained release drug delivery systems and prodrugs, examples on bioactive polymers that exhibit intrinsic therapeutic properties are relatively less. Several appealing characteristics of synthetic polymers including high molecular weight, molecular architecture, and controlled polydispersity can all be utilized to discover a new generation of therapies. For example, high molecular weight bioactive polymers can be restricted to gastrointestinal tract, where they can selectively recognize, bind, and remove target disease causing substances from the body. The appealing features of GI tract restriction and stability in biological environment render these polymeric drugs to be devoid of systemic toxicity that are generally associated with small molecule systemic drugs. The present article highlights recent developments in the rational design and synthesis of appropriate functional polymers that have resulted in a number of promising polymer based therapies and biomaterials, including some marketed products. PMID- 19682516 TI - Polymer therapeutics: clinical applications and challenges for development. PMID- 19682517 TI - XMT-1001, a novel polymeric camptothecin pro-drug in clinical development for patients with advanced cancer. AB - An overview of XMT-1001 is provided in the context of other topoisomerase I inhibitors conjugated to polymers or encapsulated in liposomes. XMT-1001 is a novel polymeric pro-drug derivative of camptothecin (CPT) with a molecular weight of 70 kDa, in which CPT is chemically tethered to a hydrophilic, biodegradable polyacetal polymer, poly(1-hydroxymethylethylene hydroxymethylformal), also called PHF or Fleximer(R). XMT-1001 releases CPT via intermediates camptothecin 20-O-(N-succinimidoglycinate) (CPT-SI), and camptothecin-20-O-(N-succinamidoyl glycinate) (CPT-SA) over an extended time period. XMT-1001 has an improved therapeutic window compared to CPT and irinotecan in human tumor xenograft models, providing a compelling rationale for clinical development. A unique feature of XMT-1001 is its dual phase release mechanism for CPT which may result in lower levels of CPT in the urine and less bladder toxicity, a serious dose limiting toxicity associated with CPT and CPT conjugated to other polymers. XMT 1001 is being evaluated in patients with advanced cancer in an ongoing Phase 1 trial. PMID- 19682518 TI - Lunasin and lunasin-like peptides inhibit inflammation through suppression of NF kappaB pathway in the macrophage. AB - Inflammation is part of the host defense mechanism against harmful matters and injury; however, aberrant inflammation is associated to the development of chronic diseases such as cancer. Lunasin is a novel peptide that demonstrates potential anticancer activity against mammalian cancer cell lines and may play a role in inflammation. The objective of this study was to determine the mechanism of action by which lunasin and lunasin-like peptides exert their anti inflammatory properties using RAW 264.7 macrophage cell line as an in vitro model. We purified three peptides (5, 8, and 14 kDa) from defatted soybean flour with a positive immunoreactivity towards lunasin mouse monoclonal antibody. Treatment with these peptides (10-50 microM) resulted in the inhibition of pro inflammatory markers in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. The 5 kDa peptide inhibited most potently pro-inflammatory markers including interleukin-6 production (IC(50)=2 microM), interleukin-1beta production (IC(50)=13 microM), nuclear factor-kappa B (NF-kappaB) transactivation (IC(50)=21 microM), cyclooxygenase-2 expression (IC(50)=25 microM), nitric oxide production (IC(50)=28 microM), inducible nitric oxide synthase expression (IC(50)=37 microM), prostaglandin E(2) production (IC(50)=41 microM), p65 nuclear translocation (IC(50)=48 microM) and p50 nuclear translocation (IC(50)=77 microM). In conclusion, lunasin and lunasin-like peptides purified from defatted soybean flour inhibited inflammation in LPS-induced RAW 264.7 macrophage by suppressing NF-kappaB pathway. PMID- 19682520 TI - Characterization of a new bioactive peptide from Potamotrygon gr. orbignyi freshwater stingray venom. AB - Brazilian freshwater stingrays, Potamotrygon gr. orbigyni, are relatively common in the middle-western regions of Brazil, where they are considered an important public health threat. In order to identify some of their naturally occurring toxin peptides available in very low amounts, we combine analytical protocols such as reversed-phase high-performance liquid chromatography (RP-HPLC), followed by a biological microcirculatory screening and mass spectrometry analysis. Using this approach, one bioactive peptide was identified and characterized, and two analogues were synthesized. The natural peptide named Porflan has the primary structure ESIVRPPPVEAKVEETPE (MW 2006.09 Da) and has no similarity with any bioactive peptide or protein found in public data banks. Bioassay protocols characterized peptides as presenting potent activity in a microcirculatory environment. The primary sequences and bioassay results, including interactions with the membrane phospholipids, suggest that these toxins are a new class of fish toxins, directly involved in the inflammatory processes of a stingray sting. PMID- 19682519 TI - Identification and expression profile of Halloween genes involved in ecdysteroid biosynthesis in Spodoptera littoralis. AB - 20-Hydroxyecdyone (20E), an active form of ecdysteroid, is the key hormone in insect growth and development. The biosynthesis of ecdysteroid is triggered and under the control of the neuropeptide, prothoracicotropic hormone (PTTH). To date, five cytochrome P450 enzymes, namely Spook (Spo), Phantom (Phm), Disembodied (Dib), Shadow (Sad) and Shade (Shd) related to ecdysteroid biosynthesis, are identified and the character of last four enzymes is well studied in Drosophila melanogaster, Bombyx mori and Manduca sexta. These genes are called Halloween genes and mediate the biosynthesis of 20E from cholesterol. In this study, we extended these works to a major pest insect in agriculture, the cotton leafworm Spodoptera littoralis (Lepidoptera: Noctuidae). We identified the sequence of five Halloween genes, and the converted amino acid sequences were compared with those of other insects. The phylogenetic analysis clearly showed separated clusters of each gene and the evolutional conservation in insects with a high similarity in Lepidoptera. Spo, phm, dib and sad were predominantly expressed in prothoracic glands, and shd was expressed in fat body and Malpighian tubules at the last instar larvae. Spo expression was kept high level between day 2 and day 4 after ecdysis. The expression of phm and dib peaked at day 2, and sad and shd expressions peaked at day 2 and day 4 after ecdysis. In addition, the hemolymph ecdysteroid titer showed a small peak at day 2 and a large peak at day 4 after ecdysis. These results suggest the importance of Halloween genes in ecdysone biosynthesis by prothoracic glands and conversion of ecdysone into 20E by fat body in larval-pupal metamorphosis. PMID- 19682521 TI - cANF causes endothelial cell hyperpolarization by activation of chloride channels. AB - OBJECTIVES: Natriuretic peptides bind with natriuretic peptide receptor (NPR)-C, which can alter cellular function through its interaction with the G(i) protein complex. NPR-C has been found to mediate the activation of K(+) channels and non selective cation channels in vascular smooth muscle and cardiac fibroblast cells, respectively. However, the electrophysiological effect of NPR-C activation on endothelial cells (EC) has not been previously examined. In this study we sought to elucidate the effect of cANF(4-23), a selective NPR-C ligand, on EC membrane potential (E(m)). METHODS/RESULTS: Changes in EC E(m) was measured through non invasive fluorescence imaging. EC were preincubated in the potentiometric dye, DiBAC(4)(3) and subsequently exposed to cANF(4-23), in the presence of selective inhibitors of ion-channels or second messengers. NPR-C expression in rat lung microvascular endothelial cells was assessed by RT-PCR. cANF(4-23) induced a sustained decrease in EC cellular fluorescence, indicating endothelial cell hyperpolarization. The cANF-induced hyperpolarization could not be attenuated by TEA, barium, ouabain or by the reduction of extracellular Ca(2+). Further, the cANF-induced hyperpolarization was insensitive to inhibition of G(i) and protein kinase G (PKG), downstream messengers of NPRs. However, the Cl(-) channel inhibitors, 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid, niflumic acid, and hypertonic saline attenuated the cANF-induced hyperpolarization. Perforated patch clamp recordings confirmed the cANF-induced current was carried by Cl(-) and could be inhibited by niflumic acid. RT-PCR confirmed expression of NPR-C in vascular smooth muscle cells but not in EC. CONCLUSIONS: cANF causes hyperpolarization that is most likely mediated via activation of Cl(-) channels by a PKG and G(i) independent mechanism. PMID- 19682522 TI - Inhibiting the inhibitors: retro-inverso Smac peptides. AB - Resistance against apoptosis-inducing anti-cancer drugs remains a severe problem in therapy. One reason is the overexpression of inhibitors of apoptosis proteins (IAPs), a group of proteins responsible for the prevention of apoptosis induction by inactivation of initiator caspases. The natural inhibitor of the IAPs is the protein Smac, which impedes the binding to the caspases. Although Smac is a potent inhibitor, Smac peptides are not very stable in vivo and thus not applicable in therapy. Bioinformatical methods were applied to design Smac derived peptides to break the therapy resistance in IAP high-expressing tumor cells. The exchange of amino acids in the Smac peptides AVPI and AVPF against unnatural amino acids leads to an improvement of the apoptosis sensitivity. The variety of Smac peptides was filtered by computational docking. Moreover, Smac derived peptides with sufficient binding to the IAPs were tested in IAP expressing Hodgkin Lymphoma cell lines. PMID- 19682523 TI - Casein phosphopeptides promote calcium uptake and modulate the differentiation pathway in human primary osteoblast-like cells. AB - Casein phosphopeptides (CPPs), originating by in vitro and/or in vivo casein digestion, are characterized by the ability to complex and solubilize calcium ions preventing their precipitation. Previous works demonstrated that CPPs improve calcium uptake by human differentiated intestinal tumor cell lines, are able to re-mineralize carious lesions in a dental enamel, and, as components of a diet, affect bone weight and calcium content in rats. The aim of the present study was to evaluate if CPPs can directly modulate bone cells activity and mineralization. Primary human osteoblast-like cells were established in culture from trabecular bone samples obtained from waste materials during orthopedic surgery. Commercial mixtures of bovine casein phosphopeptides were used. The CPP dependent intracellular calcium rises were monitored at the single cell level through fura2-fluorescence assays. Results show that CPPs: (i) stimulate calcium uptake by primary human osteoblast-like cells; (ii) increase the expression and activity of alkaline phosphatase, a marker of human osteoblast differentiation; (iii) affect the cell proliferation rate and the apoptotic level; (iv) enhance nodule formation by human SaOS-2. Taken together these results confirm the possibility that CPPs play a role as modulator of bone cell activity, probably sustained by their ability as calcium carriers. Although the exact mechanism by which CPPs act remains not completely clarified, they can be considered as potential anabolic factors for bone tissue engineering. PMID- 19682524 TI - Characterization of two novel tritiated radioligands for labelling Neuropeptide FF (NPFF(1) and NPFF(2)) receptors. AB - The binding characteristics of [(3)H]-NPVF and [(3)H]-EYF, the two first tritiated probes for the respective labelling of NPFF(1) and NPFF(2) receptors, are presented. In membranes from CHO cells transfected with the human NPFF(1) receptor, [(3)H]-NPVF labelled one class of binding sites with a high affinity (Bmax=4pmol/mg protein, Kd=2.65nM). In membranes from CHO cells transfected with the human NPFF(2) receptor, [(3)H]-EYF labelled one class of binding sites with a high affinity (Bmax=16pmol/mg protein, Kd=0.54nM). Both radioligands exhibited time-dependent binding, low (10-20%) non-specific binding and poor cross reactivity towards the related receptor subtype. The potency of different NPFF ligands to displace [(3)H]-NPVF and [(3)H]-EYF binding profiles was in good agreement with the profile previously measured by using (125)I-probes (NPFF(1) receptor: NPVF> or =1DMe=SPA-NPFF>NPFF=SQA-NPFF=QFW-NPSF>NPSF>RF9; NPFF(2) receptor: SPA-NPFF>>SQA-NPFF=QFW-NPSF=1DMe=NPFF>>NPSF=NPVF>RF9). Therefore, [(3)H]-NPVF and [(3)H]-EYF are new valuable tools for performing binding on NPFF receptors. PMID- 19682526 TI - Oxidative variables and antioxidant enzymes activities in the mdx mouse brain. AB - Dystrophin is a protein found at the plasmatic membrane in muscle and postsynaptic membrane of some neurons, where it plays an important role on synaptic transmission and plasticity. Its absence is associated with Duchenne's muscular dystrophy (DMD), in which cognitive impairment is found. Oxidative stress appears to be involved in the physiopathology of DMD and its cognitive dysfunction. In this regard, the present study investigated oxidative parameters (lipid and protein peroxidation) and antioxidant enzymes activities (superoxide dismutase and catalase) in prefrontal cortex, cerebellum, hippocampus, striatum and cortex tissues from male dystrophic mdx and normal C57BL10 mice. We observed (1) reduced lipid peroxidation in striatum and protein peroxidation in cerebellum and prefrontal cortex; (2) increased superoxide dismutase activity in cerebellum, prefrontal cortex, hippocampus and striatum; and (3) reduced catalase activity in striatum. It seems by our results, that the superoxide dismutase antioxidant mechanism is playing a protective role against lipid and protein peroxidation in mdx mouse brain. PMID- 19682525 TI - Regional brain metabolism with cytochrome c oxidase histochemistry in a PS1/A246E mouse model of autosomal dominant Alzheimer's disease: correlations with behavior and oxidative stress. AB - Mitochondrial dysfunction and brain metabolic alteration are early neurofunctional aspects in Alzheimer's disease (AD). Regional brain metabolism was analyzed by cytochrome c oxidase (COX) histochemistry in PS1-A246E mouse mutants, a model of autosomal dominant AD overexpressing beta-amyloid (Abeta) peptide without amyloidosis or cell degeneration. Immunohistochemical samples were analyzed on adjacent sections for regional Abeta1-42 levels, as well as DNA oxidative damage with 8-hydroxy-2-deoxyguanosine (8-OHdG). COX activity increased in the basal forebrain nuclear complex, specific parts of the amygdala and hippocampus, as well as in striatum and connected regions. On the contrary, a hypometabolism was observed in midline thalamic, interpeduncular, and pedonculopontine nuclei. The integration of these regions in circuitries subserving emotions, arousal, and cognitive functions may explain why neurochemical alterations in specific brain regions were linearly correlated with psychomotor slowing and disinhibition previously reported in the mutant. As the PS1-A246E model appears to mimick prodromal AD, the results support the existence of mitochondrial abnormalities prior to AD-related cognitive deficits. However, since affected PS1-A246E brain regions were not primarily those altered in AD associated histopathological features and did not systematically display either Abeta overexpression or higher 8-OHdG immunolabelling, the hypermetabolism observed seems to comprise a compensatory reaction to early mitochondrial abnormalities; furthermore, neuronal synaptic function should be considered as particularly relevant in COX activity changes. PMID- 19682527 TI - Homotypic T-cell/T-cell interaction induces T-cell activation, proliferation, and differentiation. AB - Activated CD4 T cells might induce T-cell activation from CD4 resting T cells in the absence of antigen presenting cells through interaction of activation-induced surface molecules (e.g., CD80, CD86, CD70, major histocompatibility complex class II) and their ligands constitutively expressed on resting T cells. Supporting this hypothesis, CD4 memory T cells proliferated in response to contact with activated T cells and expressed activation markers, such as CD25, CD30, and CD69. Analysis of their cytokine profile revealed differentiation of interleukin (IL) 10 and interferon-gamma double-producing cells in response to contact with activated T helper (Th) 1 effector cells, and interleukin (IL)-4-producing cells in response to contact with activated Th2 effector cells. Whereas neutralization of interferon-gamma or IL-4 during co-culture did not diminish the frequency of the arising cytokine-producing cells, separation of the responder cells from effector cells significantly decreased cytokine secretion. Specific blocking of particular receptor/ligand interactions denoted above could not prevent cytokine production induced by T-cell/T-cell interaction. However, blockade of all of the receptor/counterreceptor pairs significantly inhibited cytokine production, although not completely. Given the immunomodulatory capacity of IL-4 and IL-10, these findings might indicate a novel contact dependent negative feedback mechanism to control T-cell-driven immunity. PMID- 19682528 TI - Safety assessment of a modified acetolactate synthase protein (GM-HRA) used as a selectable marker in genetically modified soybeans. AB - Acetolactate synthase (ALS) enzymes have been isolated from numerous organisms including soybeans (Glycine max; GM-ALS) and catalyze the first common step in biosynthesis of branched chain amino acids. Expression of an ALS protein (GM-HRA) with two amino acid changes relative to native GM-ALS protein in genetically modified soybeans confers tolerance to herbicidal active ingredients and can be used as a selectable transformation marker. The safety assessment of the GM-HRA protein is discussed. Bioinformatics comparison of the amino acid sequence did not identify similarities to known allergenic or toxic proteins. In vitro studies demonstrated rapid degradation in simulated gastric fluid (<30s) and intestinal fluid (<1min). The enzymatic activity was completely inactivated at 50 degrees C for 15 min demonstrating heat lability. The protein expressed in planta is not glycosylated and genetically modified soybeans expressing the GM-HRA protein produced similar protein/allergen profiles as its non-transgenic parental isoline. No adverse effects were observed in mice following acute oral exposure at a dose of at least 436 mg/kg of body weight or in a 28-day repeated dose dietary toxicity study at doses up to 1247 mg/kg of body weight/day. The results demonstrate GM-HRA protein safety when used in agricultural biotechnology. PMID- 19682529 TI - Reaction on Gargas et al.: Acrylamide: consideration of species differences and nonlinear processes in estimating risk and safety for human ingestion. PMID- 19682530 TI - The benchmark dose approach in food risk assessment: is it applicable and worthwhile? AB - The benchmark dose (BMD) approach is being increasingly used in the area of food risk assessment because it offers several advantages compared to the conventional no-observed-adverse-effect-level approach. The aim of this work was to check the applicability of the BMD approach on toxicity data available from pesticides, mycotoxins and natural toxins. Based on toxicological evaluations, the pivotal study was identified. Detailed data from the original study were retrieved and used for BMD modelling. Twenty-five studies used for BMD modelling were analysed with regard to study design: total number of animals, number of dose levels, and spacing between dose levels. The quality of the modelled endpoints was evaluated according to the following aspects: BMD/BMDL ratio, test for goodness of fit and BMD in the range of dose levels. If one of these aspects was not fulfilled, the BMD derived from this endpoint was considered to be uncertain to some extent and corresponding modelled data sets were examined. The present work demonstrates that the BMD approach is in principle applicable to pesticides, mycotoxins, and natural toxins. Although large differences relating to data availability and data quality were noticed, 69 of 82 modelled endpoints (84%) fulfilled the three quality aspects of BMD modelling. PMID- 19682531 TI - Harmonisation of food consumption data format for dietary exposure assessments of chemicals analysed in raw agricultural commodities. AB - In this paper, we present an approach to format national food consumption data at raw agricultural commodity (RAC) level. In this way, the data is both formatted in a harmonised way given the comparability of RACs between countries, and suitable to assess the dietary exposure to chemicals analysed in RACs at a European level. In this approach, consumption data needs to be converted to edible part of RAC (e-RAC) level using a RAC conversion database. To subsequently use this data in exposure assessments, both e-RACs and RACs analysed in chemical control programmes should be classified via a uniform system. Furthermore, chemical concentrations in RACs may need to be converted to e-RAC level using processing factors. To illustrate the use of this approach, we describe how the Dutch RAC conversion database was used to convert consumption data of four national consumption surveys to e-RAC level, and the use of the FAO/WHO Codex Classification system of Foods and Animal Feeds to harmonise the classification. We demonstrate that this approach works well for pesticides and glycoalkaloids, and is an essential step forward in the harmonisation of risk assessment procedures within Europe when addressing chemicals analysed in RACs by all national food control systems. PMID- 19682532 TI - Disinfectant properties of essential oils from Salvia officinalis L. cultivated in Tunisia. AB - The essential oils were obtained by hydro-distillation of the aerial part of Salvia officinalis L. cultivated in Sfax gardens, Tunisia. The obtained oils were analyzed by gas chromatography-mass spectrometry (GC-MS) and 44 compounds were identified. Strong bactericidal and fungicidal effects were shown using the NCCLS broth dilution, EN 1275 and EN 1276 standard methods. The minimal cidal concentrations (MCCs) values ranged from 0.031 to 0.25 microLmL(-1). The essential oils concentrations of 0.5% and 1% (v/v) resulted in a reduction in viability higher than 5 and 4 log units per mL for the standard bacteria and fungi, respectively, within a contact time of 5 min. Using an air sampler and an aroma dispenser, vaporisation of 0.25 mLm(-3) of S. officinalis essential oils resulted in (72%, 73% and 70%) and (54%, 55% and 55%) reduction of the total microbial count and the total count of yeasts and moulds, after a residence time of 1h, 6h and 24h in a selected testing room, respectively. S. officinalis essential oils showed a potent vapour activity against a panel of bacteria, yeasts and fungi. This supported their use as a natural eco-friendly disinfectant to manage airborne microbes. PMID- 19682534 TI - Adapting the diagnostic definitions of the RDC/TMD to routine clinical practice: a feasibility study. AB - The Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) is a well-known diagnostic tool for clinical trials on TMD. OBJECTIVES: This study aims to assess the reliability, validity and feasibility of a new method of physically diagnosing temporomandibular disorders (TMD), designed for routine clinical use. This version, known as Clinical Examination Protocol-TMD (CEP-TMD), was compared to the gold standard original RDC/TMD. METHODS: A total of 49 subjects (41 referred TMD patients and 8 symptom free subjects) were examined using both RDC/TMD and CEP-TMD versions. Three examiners, with varying levels of experience in diagnosing TMD, worked in pairs. Each member of a pair saw the same patient twice, once for the RDC/TMD and once for the CEP-TMD examination. The examination order was randomized. Each patient's examinations alternated between examiners to reduce the memory effect. Examinations could yield single, multiple or no diagnosis. Kappa statistics were calculated to estimate reliability. RESULTS: There was substantial overall agreement between the CEP-TMD and the RDC/TMD (kappa=0.70). Intra-examination agreements were substantial in both RDC/TMD (kappa=0.70) and CEP-TMD (kappa=0.90). For examination and diagnosis, the CEP-TMD was almost 3 min faster than the RDC/TMD (p<0.05). CONCLUSIONS: It was concluded that the CEP-TMD's diagnosis is comparable to the RDC/TMD thus providing a convenient and intuitive approach for dentists to physically diagnose TMD in clinical practice. The well-established RDC/TMD remains the gold standard for research diagnosis of TMD. PMID- 19682533 TI - Inflammatory gene expression in response to sub-lethal ricin exposure in Balb/c mice. AB - The toxin ricin has been shown to cause inflammatory lung damage, leading to pulmonary oedema and, at higher doses, mortality. In order to understand the genetic basis of this inflammatory cascade a custom microarray platform (1509 genes) directed towards immune and inflammatory markers was used to investigate the temporal expression profiles of genes in a Balb/c mouse model of inhalational ricin exposure. To facilitate examination of those genes involved in both inflammatory cascades and wound repair the dose which was investigated was sub lethal across a 96-h time course. Histopathology of the lung was mapped across the time course and genetic responses considered in the context of overall lung pathology. Six hundred and eighty-five genes were found to be statistically significantly different compared to controls, across the time course and these genes have been investigated in the context of their biological function in ricin poisoning. As well as confirming key inflammatory markers associated with ricin intoxication (TNFalpha and IL1beta) several pathways that are altered in expression were identified following pulmonary exposure to ricin. These genes included those involved in cytokine-cytokine signalling cascades (IL1, IL1r, IL1r2, Ccl 4, 6, 10), focal adhesion (Fn1, ICAM1) and tissue remodelling (VEGF, Pim1). Furthermore, the observed alteration in expression of matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) indicates a key role in membrane integrity and cellular adhesion in ricin poisoning. Data captured using this transcriptomic approach could be used to develop a specific approach to the treatment of inhalational ricin exposure. This work was conducted as part of a wider programme of work to compare a number of militarily relevant lung damaging agents, with a view to establishing a rational basis for the identification of more generic medical countermeasures. PMID- 19682535 TI - Synchrony of spatial populations induced by colored environmental noise and dispersal. AB - Spatial synchrony of oscillating populations has been observed in many ecological systems, and its influences and causes have attracted the interest of ecologists. Spatially correlated environmental noises, dispersal, and trophic interactions have been considered as the causes of spatial synchrony. In this study, we develop a spatially structured population model, which is described by coupled map lattices and incorporates both dispersal and colored environmental noise. A method for generating time series with desired spatial correlation and color is introduced. Then, we use these generated time series to analyze the influence of noise color on synchrony in population dynamics. The noise color refers to the temporal correlation in the time series data of the noise, and is expressed as the degree of (first-order) autocorrelation for autoregressive noise. Patterns of spatial synchrony are considered for stable, periodic and chaotic population dynamics. Numerical simulations verify that environmental noise color has a major influence on the level of synchrony, which depends strongly on how noise is introduced into the model. Furthermore, the influence of noise color also depends on patterns of dispersal between local populations. In addition, the desynchronizing effect of reddened noise is always weaker than that of white noise. From our results, we notice that the role of reddened environmental noise on spatial synchrony should be treated carefully and cautiously, especially for the spatially structured populations linked by dispersal. PMID- 19682536 TI - Study of the transport of thyroid hormone by transporters of the SLC10 family. AB - Transport of (sulfated) iodothyronines across the plasma membrane is required for their intracellular metabolism. Rat Na(+)/taurocholate cotransporting polypeptide (Ntcp; Slc10a1) has been identified as an important transporter protein. We demonstrate that among the 7 members of the solute carrier family SLC10, only human SLC10A1 mediates sodium-dependent transport of the iodothyronine T4 and iodothyronine sulfates T3S and T4S. In contrast to SLC10A2-7, cells co-expressing SLC10A1 and the deiodinase D1 demonstrate a dramatic increase in T3S and T4S metabolism. The SLC10A1 substrates taurocholate, DHEAS and E3S inhibit T3S and T4S transport. Furthermore, co-transfection of SLC10A1 with CRYM, a well-known intracellular iodothyronine-binding protein, results in an enhanced intracellular accumulation of T3S and T4S, indicating that CRYM binds iodothyronine sulfates. The present findings indicate that the liver-specific transporter SLC10A1 transports (sulfated) iodothyronines, thereby increasing their intracellular availability. Therefore, SLC10A1 may fulfill a critical step in providing liver D1 with iodothyronine sulfates for rapid degradation. PMID- 19682537 TI - Expression of adipokines and estrogen receptors in adipose tissue and placenta of patients with gestational diabetes mellitus. AB - The purpose of this study was to assess the expression profile of genes with potential role in the development of insulin resistance (adipokines, cytokines/chemokines, estrogen receptors) in subcutaneous adipose tissue (SAT), visceral adipose tissue (VAT) and placenta of pregnant women with gestational diabetes mellitus (GDM) and age-matched women with physiological pregnancy at the time of Caesarean section. qRT-PCR was used for expression analysis of the studied genes. Leptin gene expression in VAT of GDM group was significantly higher relative to control group. Gene expressions of interleukin-6 and interleukin-8 were significantly increased, whereas the expressions of genes for estrogen receptors alpha and beta were significantly reduced in SAT of GDM group relative to controls, respectively. We found no significant differences in the expression of any genes of interest (LEP, RETN, ADIPOR1, ADIPOR2, TNF-alpha, CD68, IL-6, IL-8, ER alpha, ER beta) in placentas of women with GDM relative to controls. We conclude that increased expression of leptin in visceral adipose depot together with increased expressions of proinflammatory cytokines and reduced expressions of estrogen receptors in subcutaneous fat may play a role in the etiopathogenesis of GDM. PMID- 19682539 TI - Adipokine dysregulation, adipose tissue inflammation and metabolic syndrome. AB - Obesity plays a causative role in the pathogenesis of the metabolic syndrome. Adipokines may link obesity to its co-morbidities. Most adipokines with pro inflammatory properties are overproduced with increasing adiposity, while some adipokines with anti-inflammatory or insulin-sensitizing properties, like adiponectin are decreased. This dysregulation of adipokine production may promote obesity-linked metabolic disorders and cardiovascular disease. Besides considering adipokines, this review will also highlight the cellular key players and molecular mechanisms involved in adipose inflammation. Targeting the changes in the cellular composition of adipose tissue, the underlying molecular mechanisms, and the altered production of adipokines may have therapeutic potential in the management of the metabolic syndrome. PMID- 19682538 TI - Regulation of spermatogenesis in the microenvironment of the seminiferous epithelium: new insights and advances. AB - Spermatogenesis is a complex biochemical event, involving the participation of the hypothalamus and the pituitary gland via secretion of the hypothalamus hormone GnRH, and two pituitary hormones FSH and LH. Thus, the hypothalamic pituitary-testicular axis is a crucial regulatory axis for testicular function. Recent studies have shown that in the microenvironment of the seminiferous epithelium, wherein each Sertoli cell supports approximately 30-50 germ cells at different stages of development, locally produced autocrine and paracrine factors are also involved in spermatogenesis, in particular at the level of cell junctions. These cell junctions at the Sertoli-Sertoli and Sertoli-germ cell interface are crucial for coordinating different events of spermatogenesis by sending signals back-and-forth between Sertoli and germ cells, in order to precisely regulate spermatogonial cell renewal by mitosis, cell cycle progression, meiosis, spermiogenesis, germ cell movement across the epithelium, spermiation and germ cell apoptosis. In this minireview, we provide an update on these latest findings for an emerging new concept regarding the presence of a local "apical ectoplasmic specialization-blood-testis barrier hemidesmosome/basement membrane" functional axis that regulates the events of spermiation and blood-testis barrier (BTB) restructuring via paracrine/autocrine factors and polarity proteins produced locally in the seminiferous epithelium. These findings provide a new window of research for investigators in the field to tackle the functional regulation of spermatogenesis. PMID- 19682540 TI - Specific transgene expression in mouse pancreatic beta-cells under the control of the porcine insulin promoter. AB - The availability of regulatory sequences directing tissue-specific expression of transgenes in genetically modified mice and large animals is a prerequisite for the development of adequate models for human diseases. The rat insulin 2 gene (Ins2) promoter, widely used to achieve transgene expression in pancreatic beta cells of mice, also directs expression to extrapancreatic tissues and performs poorly in isolated pancreatic islets of human, mouse, and pig. To evaluate whether the full 5' untranslated region (UTR) of the porcine insulin gene (INS) confers robust and specific expression in beta-cells we generated an expression cassette containing 1500bp of the porcine INS 5' UTR and the 3' UTR of the bovine growth hormone gene (GH). The cassette was designed to allow easy exchange of the sequences to be expressed and easy removal of the vector backbone from the expression cassette. To evaluate the properties of the cassette, we initially inserted a cDNA encoding human betacellulin, a growth factor known to affect structural and functional parameters of beta-cells. After confirming the functionality and specificity of the construct in vitro, transgenic mouse lines were generated by pronuclear DNA microinjection. Using RT-PCR, immunohistochemistry and immunofluorescence, we show that transgenic mice expressed human betacellulin exclusively in beta-cells. Confirming the proposed insulinotropic effect of betacellulin, transgenic mice showed improved glucose tolerance. We conclude that the newly designed expression cassette containing 1500bp of the porcine insulin promoter 5' UTR confers robust and specific transgene expression to beta-cells in vitro and in transgenic mice. PMID- 19682541 TI - Zebrafish cyp11a1 and hsd3b genes: structure, expression and steroidogenic development during embryogenesis. AB - Zebrafish has been used increasingly as a good animal model for a number of studies. To facilitate the use of this zebrafish model, the current report put emphasis on the study of two steroidogenic genes: cyp11a1 and hsd3b. These two genes encode enzymes that catalyze the first two steps of the steroidogenic pathway, and both enzymes are important for the synthesis of all steroids. Zebrafish cyp11a1 and hsd3b genes are expressed in the same cells in the gonads and interrenal gland. The interrenal gland is the counterpart of mammalian adrenal; it is located inside the head kidney and is developed parallel to the development of the pronephros. In addition, cyp11a1 and hsd3b are also expressed in the blastomeres of the early embryos before gastrulation, and in the extra embryonic yolk syncytial layer during gastrulation. This early expression implies a novel role of steroids at gastrulation. PMID- 19682542 TI - Mediation of protein kinase C zeta in mu-opioid receptor activation for increase of glucose uptake into cultured myoblast C2C12 cells. AB - The present study is designed to investigate the role of atypical protein kinase C (PKC) in the signaling of mu-opioid receptors (MOR) for glucose uptake in myoblast C(2)C(12) cells. Loperamide enhanced the uptake of radioactive deoxyglucose into C(2)C(12) cells in a concentration-dependent manner that was abolished in cells pre-incubated with GF109203X at concentrations sufficient to block PKC. Inhibition of the atypical zeta (zeta) isoform of PKC using myristoylated PKC pseudosubstrate resulted in a concentration-dependent decrease of loperamide-stimulated glucose uptake into C(2)C(12) cells. In addition, loperamide elicited the phosphorylation of PKC-zeta in C(2)C(12) cells in a concentration-dependent manner that was abolished by pretreatment with naloxonazine at concentrations sufficient to block MOR. These results suggest the mediation of PKC-zeta in MOR signaling for glucose uptake in C(2)C(12) cells. Activation of PKC-zeta by MOR stimulation is highly relevant to the search for therapeutic targets for glucose transport in insulin-sensitive tissues. PMID- 19682543 TI - Proteomic profiling of the insoluble fractions in the rat hippocampus post propofol anesthesia. AB - Cognitive dysfunction after propofol anesthesia has been previously found. The underlying mechanisms of this sequel remain unclear. Insoluble proteins as major targets of anesthetics participated in various pathophysiological processes. This study aimed to provide evidence that changes in insoluble proteome in rat hippocampus may be involved in molecular mechanism of cognitive dysfunction following propofol anesthesia. Proteins extracted from rat hippocampus were separated by two-dimensional electrophoresis (2-DE). Their expression patterns were observed at 1, 6, 24 h and 7 days after 3 h of propofol anesthesia. Differentially expressed protein spots among groups were submitted to matrix assisted laser desorption/ionization time of flight mass spectrometer (MALDI-TOF MS) assay and peptide mass fingerprinting (PMF) identification. Identified proteins were further analyzed through Gene Ontology (GO). Results of 2-DE were selectively assayed using Western blot and RT-PCR. Fifty-nine differentially expressed proteins were detected, among which 43 were identified through MALDI TOF MS. Most identified proteins were distributed in organelles and membranes. According to biological process category, 27 proteins were involved in metabolic process, 19 in developmental process, 14 in stimulus-response, and 21 in biological regulation. Most changes took place within 24 h, with more down regulation within 6 h. Twelve proteins did not restore to the basic level until the 7th day after propofol anesthesia. Expressions of insoluble proteome dynamically changed following propofol anesthesia. Down-regulations at early stage might produce depressive effects, which may be involved in molecular mechanism of cognitive dysfunction after propofol anesthesia. PMID- 19682544 TI - Motor cortex stimulation for ALS: a double blind placebo-controlled study. AB - Preliminary data suggest that repetitive transcranial magnetic stimulation (rTMS) of the brain may produce a modest slowing of disease progression in amyotrophic lateral sclerosis (ALS). The present study was designed to test the hypothesis that rTMS given as continuous theta burst stimulation (cTBS), repeated monthly for one year, would affect ALS progression. We performed a double blind, placebo controlled trial. Twenty patients with ALS were randomly allocated to blinded real or placebo stimulation. cTBS of the motor cortex was performed for five consecutive days every month for one year. Primary outcome was the rate of decline as evaluated with the revised ALS functional rating scale (ALSFRS-R). Treatment was well tolerated. There was no significant difference in the ALSFRS-R score deterioration between patients treated with real or placebo stimulation. ALSFRS-R mean scores declined from 32.0 (SD 7.1) at study entry to 23.1 (SD 6.3) at 12 months in patients receiving real cTBS and from 31.3 (SD 6.9) to 21.2 (SD 6.0) in those receiving placebo stimulation. Although cTBS proved a safe procedure, on the basis of the present findings a larger randomized confirmatory trial seems unjustified in ALS patients, at least in advanced stage of the disease. PMID- 19682545 TI - Gamma(2)-melanocyte stimulating hormone decreases food intake in chicks. AB - The role of gamma melanocyte stimulating hormone (gamma-MSH) in appetite regulation is controversial in mammals and to our knowledge unreported within the avian class. Thus, the present study was designed to determine the effects of intracerebroventricularly (i.c.v.) administered gamma2-MSH on food intake using Cobb-500 chicks as models. In Experiment 1, chicks that received i.c.v. gamma2 MSH decreased their food intake throughout the 180 min observation period and plasma glucose concentration was not affected. Water intake was also decreased in i.c.v. gamma2-MSH-treated chicks, but only from 30 to 90 min post-injection. In Experiment 2, food pecking efficiency was decreased in i.c.v. gamma2-MSH-treated chicks and the amount of time spent sitting was increased. Other behaviors were not significantly affected by i.c.v. gamma2-MSH including distance traveled, the number of jumps, escape attempts, defecations, food pecks, exploratory pecks, and the amount of time spent standing, preening, perching, or in deep rest. These data suggest that gamma2-MSH is associated with anorexigenic effects and because of gamma-MSH's selectivity, implicates the melanocortin 3 receptor in appetite regulation. PMID- 19682546 TI - NeuN is not a reliable marker of dopamine neurons in rat substantia nigra. AB - Quantification of neuronal cell number is a key endpoint in the characterization of neurodegenerative disease models and neuroprotective regimens. Immunohistochemistry for phenotypic markers, followed by unbiased stereology is often used to quantify the relevant neuronal population. To control for loss of phenotypic markers in the absence of cell death, or to determine if other types of neurons are lost, a general neuronal marker is often desired. Vertebrate neuron-specific nuclear protein (NeuN) is reportedly expressed in most mammalian neurons. In Parkinson's disease models, NeuN has been widely used to determine if there is actual nigral dopamine neuron loss or simply loss of tyrosine hydroxylase expression, a prominent phenotypic marker. To date, the qualitative value of NeuN expression as such a marker in the substantia nigra has not been assessed. Midbrain tissue sections from control rats were stained for NeuN and tyrosine hydroxylase and assessed by light or confocal microscopy. Here we report that NeuN expression level in the rat substantia nigra was highly variable, with many faintly stained cells that would not be meet stereological scoring criteria. Additionally, dopamine neurons with little or no NeuN expression were readily identified. Subcellular compartmentalization of NeuN expression was also variable, with many cells dorsal and ventral to the nigra exhibiting expression in both the nucleus and cytoplasm. NeuN expression also appeared to be much higher in non-dopamine neurons within the ventral midbrain. This characterization of nigral NeuN expression suggests that it is not useful as a quantitative general neuronal marker in the substantia nigra. PMID- 19682547 TI - Modification of cortical excitability in neuropathic rats: a voltage-sensitive dye study. AB - Recent advances in optical imaging techniques have made it possible to monitor neural activity and provided powerful tools to reveal the spatiotemporal patterns of neural activity. We used optical imaging to determine whether nerve injury affects excitability of the sensory cortex. Male Sprague-Dawley rats were subjected to neuropathic surgery consisting of a tight ligation and transection of the left tibial and sural nerves while under pentobarbital anesthesia. The rats were reanesthetized with urethane two weeks post-operatively, and the exposed cortex surfaces were stained with a voltage-sensitive dye (di-2-ANEPEQ). After electrical stimulation of the receptive field, optical signals from the cerebral cortex were recorded using an optical imaging system. Increased optical intensity and an enlarged area of activation were observed in the cerebral cortex of neuropathic rats during electrical stimulation compared to normal or sham operated rats. Higher electric stimulation resulted in more intensity and a larger area of activation in neuropathic rats. These results suggest that cortical excitability, resulting from peripheral stimulation, may be affected by nerve injury, which indicates a degree of neural plasticity. PMID- 19682548 TI - Effects of task complexity and sensory conflict on goal-directed movement. AB - Interactions between brain regions are necessary for compound activities to take place. Accordingly, evaluating hemispheric information processing during skilled behaviour provides valuable knowledge about brain regulation. To this end, the present study assessed the neural changes in response to task complexity and visuomotor discrepancy during motor (drawing) actions. EEG phase synchronization, expressing interregional communication, showed that visuomotor discordance perturbed information processing across both hemispheres, whereas task complexity induced pronounced adjustments in the left (dominant) hemisphere. However, the effects of task complexity and sensorimotor conflict interacted, and suggested that the main process of spatiotemporal integration was localized within the left hemisphere. Furthermore, a significant association between left hemisphere couplings and performance accuracy proposed that connectivity strength and behavioural output are linked with one another. These results suggest that functional connectivity patterns provide higher-order associations for information coding during skilled actions. PMID- 19682549 TI - Transcriptional and post-transcriptional regulation of mitochondrial biogenesis in skeletal muscle: effects of exercise and aging. AB - Acute contractile activity of skeletal muscle initiates the activation of signaling kinases. This promotes the phosphorylation of transcription factors, leading to enhanced DNA binding and transcriptional activation and/or repression. The mRNA products of nuclear genes encoding mitochondrial proteins are translated in the cytosol and imported into pre-existing mitochondria. When contractile activity is repeated, the recapitulation of these cellular events progressively leads to an expansion of the mitochondrial reticulum within muscle. This has physiologically relevant health benefit, including enhanced lipid metabolism and reduced muscle fatigability. In aging skeletal muscle, the response to contractile activity appears to be attenuated, suggesting that a greater contractile stimulus is required to attain a similar phenotype adaptation. This review summarizes our current understanding of the effects of exercise on the gene expression pathway leading to organelle biogenesis in muscle. PMID- 19682550 TI - Molecular pathology of RUNX3 in human carcinogenesis. AB - A major goal of molecular biology is to elucidate the mechanisms underlying cancer development and progression in order to achieve early detection, better diagnosis and staging and novel preventive and therapeutic strategies. We feel that an understanding of Runt-related transcription factor 3 (RUNX3)-regulated biological pathways will directly impact our knowledge of these areas of human carcinogenesis. The RUNX3 transcription factor is a downstream effector of the transforming growth factor-beta (TGF-beta) signaling pathway, and has a critical role in the regulation of cell proliferation and cell death by apoptosis, and in angiogenesis, cell adhesion and invasion. We previously identified RUNX3 as a major gastric tumor suppressor by establishing a causal relationship between loss of function and gastric carcinogenesis. More recently, we showed that RUNX3 functions as a bona fide initiator of colonic carcinogenesis by linking the Wnt oncogenic and TGF-beta tumor suppressive pathways. Apart from gastric and colorectal cancers, a multitude of epithelial cancers exhibit inactivation of RUNX3, thereby making it a putative tumor suppressor in human neoplasia. This review highlights our current understanding of the molecular mechanisms of RUNX3 inactivation in the context of cancer development and progression. PMID- 19682551 TI - Ion beam radiobiology and cancer: time to update ourselves. AB - High-energy protons and carbon ions exhibit an inverse dose profile allowing for increased energy deposition with penetration depth. Additionally, heavier ions like carbon beams have the advantage of a markedly increased biological effectiveness characterized by enhanced ionization density in the individual tracks of the heavy particles, where DNA damage becomes clustered and therefore more difficult to repair, but is restricted to the end of their range. These superior biophysical and biological profiles of particle beams over conventional radiotherapy permit more precise dose localization and make them highly attractive for treating anatomically complex and radioresistant malignant tumors but without increasing the severe side effects in the normal tissue. More than half a century since Wilson proposed their use in cancer therapy, the effects of particle beams have been extensively investigated and the biological complexity of particle beam irradiation begins to unfold itself. The goal of this review is to provide an as comprehensive and up-to-date summary as possible of the different radiobiological aspects of particle beams for effective application in cancer treatment. PMID- 19682552 TI - Tumor cell energy metabolism and its common features with yeast metabolism. AB - During the last decades a considerable amount of research has been focused on cancer. A number of genetic and signaling defects have been identified. This has allowed the design and screening of a number of anti-tumor drugs for therapeutic use. One of the main challenges of anti-cancer therapy is to specifically target these drugs to malignant cells. Recently, tumor cell metabolism has been considered as a possible target for cancer therapy. It is widely accepted that tumors display an enhanced glycolytic activity and oxidative phosphorylation down regulation (Warburg effect). Therefore, it seems reasonable that disruption of glycolysis might be a promising candidate for specific anti-cancer therapy. Nonetheless, the concept of aerobic glycolysis as the paradigm of tumor cell metabolism has been challenged, as some tumor cells use oxidative phosphorylation. Mitochondria are of special interest in cancer cell energy metabolism, as their physiology is linked to the Warburg effect. Besides, their central role in apoptosis makes these organelles a promising "dual hit target" for selectively eliminate tumor cells. Thus, it is desirable to have an easy-to use and reliable model in order to do the screening for energy metabolism inhibiting drugs to be used in cancer therapy. From a metabolic point of view, the fermenting yeast Saccharomyces cerevisiae and tumor cells share several features. In this paper we will review these common metabolic properties and we will discuss the possibility of using S. cerevisiae as an early screening test in the research for novel anti-tumor compounds used for the inhibition of tumor cell metabolism. PMID- 19682554 TI - Aggrecan-based extracellular matrix shows unique cortical features and conserved subcortical principles of mammalian brain organization in the Madagascan lesser hedgehog tenrec (Echinops telfairi Martin, 1838). AB - The Madagascan tenrecs (Afrotheria), an ancient mammalian clade, are characterized by unique brain anatomy. Striking features are an expanded paleocortex but a small and poorly differentiated neocortex devoid of a distinct granular layer IV. To investigate the organization of cortical areas we analyzed extracellular matrix components in perineuronal nets (PNs) using antibodies to aggrecan, lectin staining and hyaluronan-binding protein. Selected subcortical regions were studied to correlate the cortical patterns with features in evolutionary conserved systems. In the neocortex, paleocortex and hippocampus PNs were associated with nonpyramidal neurons. Quantitative analysis in the cerebral cortex revealed area-specific proportions and laminar distribution patterns of neurons ensheathed by PNs. Cortical PNs showed divergent structural phenotypes. Diffuse PNs forming a cotton wool-like perisomatic rim were characteristic of the paleocortex. These PNs were associated with a dense pericellular plexus of calretinin-immunoreactive fibres. Clearly contoured PNs were devoid of a calretinin-positive plexus and predominated in the neocortex and hippocampus. The organization of the extracellular matrix in subcortical nuclei followed the widely distributed mammalian type. We conclude that molecular properties of the aggrecan-based extracellular matrix are conserved during evolution of mammals; however, the matrix scaffold is adapted to specific wiring patterns of cortical and subcortical neuronal networks. PMID- 19682553 TI - Rapamycin protects against rotenone-induced apoptosis through autophagy induction. AB - Ubiquitin proteasome system (UPS) and autophagy lysosome pathway (ALP) are the two most important routes for degradation of aggregated/misfolded proteins. Additionally, ALP is so far the only known route to clear entire organelles, such as mitochondria. We proposed that enhancement of ALP may be beneficial for some neurodegenerative disorders, such as Parkinson's disease (PD), in which the accumulation of aggregated/misfolded proteins and the dysfunction of mitochondria are the two major pathogenesis. Mitochondrial complex I inhibitor rotenone, which causes dysfunction mitochondria and UPS, has been considered as one of the neurotoxins related to PD. In this study, rotenone-exposed human neuronal SH-SY5Y cells were used as an in vitro model for us to determine whether autophagy enhancer rapamycin could protect against rotenone-induced injury and its underlying mechanisms. The observed results showed that rapamycin alleviated rotenone-induced apoptosis, whose effects were partially blocked when autophagy related gene 5 (Atg5) was suppressed by Atg5 small interference RNA (siRNA) transfection. Additionally, the results showed that rapamycin pretreatment diminished rotenone-induced accumulation of high molecular weight ubiquitinated bands, and reduced rotenone-induced increase of cytochrome c in cytosolic fraction and decreased mitochondrial marker cytochrome oxidase subunit IV (COX IV) in mitochondrial fraction. The changes in cytochrome c and COX IV indicated that the decreased translocation of cytochrome c from mitochondria to cytosol was probably due to the turn over of entire injured mitochondria. The results that lysosome and mitochondria were colocolized within the cells pretreated with rapamycin and that the mitochondria could be found within autophagy double membrane structures further supported that the damaged mitochondria might be cleared through autophagy, which process has been termed as "mitophagy." Our studies suggested that autophagy enhancer rapamycin is neuroprotective against rotenone-induced apoptosis through autophagy enhancement. We concluded that pharmacologically induction of autophagy by rapamycin may represent a useful therapeutic strategy as disease-modifiers in PD. PMID- 19682555 TI - Aquaporin-4 in brain and spinal cord oedema. AB - Brain oedema is a major clinical problem produced by CNS diseases (e.g. stroke, brain tumour, brain abscess) and systemic diseases that secondarily affect the CNS (e.g. hyponatraemia, liver failure). The swollen brain is compressed against the surrounding dura and skull, which causes the intracranial pressure to rise, leading to brain ischaemia, herniation, and ultimately death. A water channel protein, aquaporin-4 (AQP4), is found in astrocyte foot processes (blood-brain border), the glia limitans (subarachnoid cerebrospinal fluid-brain border) and ependyma (ventricular cerebrospinal fluid-brain border). Experiments using mice lacking AQP4 or alpha syntrophin (which secondarily downregulate AQP4) showed that AQP4 facilitates oedema formation in diseases causing cytotoxic (cell swelling) oedema such as cerebral ischaemia, hyponatraemia and meningitis. In contrast, AQP4 facilitates oedema elimination in diseases causing vasogenic (vessel leak) oedema and therefore AQP4 deletion aggravates brain oedema produced by brain tumour and brain abscess. AQP4 is also important in spinal cord oedema. AQP4 deletion was associated with less cord oedema and improved outcome after compression spinal cord injury in mice. Here we consider the possible routes of oedema formation and elimination in the injured cord and speculate about the role of AQP4. Finally we discuss the role of AQP4 in neuromyelitis optica (NMO), an inflammatory demyelinating disease that produces oedema in the spinal cord and optic nerves. NMO patients have circulating AQP4 IgG autoantibody, which is now used for diagnosing NMO. We speculate how NMO-IgG might produce CNS inflammation, demyelination and oedema. Since AQP4 plays a key role in the pathogenesis of CNS oedema, we conclude that AQP4 inhibitors and activators may reduce CNS oedema in many diseases. PMID- 19682557 TI - The effect of orientation adaptation on responses of lateral geniculate nucleus neurons with high orientation bias in cats. AB - Adaptation to stimulus orientation is assumed to have a cortical basis, but few studies have addressed whether it affects the activity of subcortical neurons. Using single-unit recording, we studied the effects of orientation adaptation on the responses of lateral geniculate nucleus (LGN) neurons with high orientation bias (OB) in anesthetized and paralyzed cats. Following adaptation to one stimulus orientation, the response at the adapting orientation was decreased, and the preferred orientation was shifted away from the adapting orientation. This phenomenon was similar to the effects observed for orientation adaptation in the primary visual cortex (V1), and was obvious when the adapting orientation was at an appropriate location relative to the original preferred orientation. Moreover, when the V1 was inactivated, the response at the adapting orientation was also decreased but the preferred orientation did not show a systematic shift after orientation adaptation in LGN. This result indicates that cortical feedback contributes to the effect of orientation adaptation on LGN neurons, which have a high OB. These data provide an example of how the corticothalamic loop modulates the processing of visual information, and suggest that the LGN is not only a simply passive relay but also a modulator of visual information. PMID- 19682556 TI - Immunocytochemical identification of proteins involved in dopamine release from the somatodendritic compartment of nigral dopaminergic neurons. AB - We examined the somatodendritic compartment of nigral dopaminergic neurons by immunocytochemistry and confocal microscopy, with the aim of identifying proteins that participate in dopamine packaging and release. Nigral dopaminergic neurons were identified by location, cellular features and tyrosine hydroxylase immunoreactivity. Immunoreactive puncta of vesicular monoamine transporter type 2 and proton ATPase, both involved in the packaging of dopamine for release, were located primarily in dopaminergic cell bodies, but were absent in distal dopaminergic dendrites. Many presynaptic proteins associated with transmitter release at fast synapses were absent in nigral dopaminergic neurons, including synaptotagmin 1, syntaxin1, synaptic vesicle proteins 2a and 2b, synaptophysin and synaptobrevin 1 (VAMP 1). On the other hand, syntaxin 3, synaptobrevin 2 (VAMP 2) and SNAP-25-immunoreactivities were found in dopaminergic somata and dendrites Our data imply that the storage and exocytosis of dopamine from the somatodendritic compartment of nigral dopaminergic neurons is mechanistically distinct from transmitter release at axon terminals utilizing amino acid neurotransmitters. PMID- 19682558 TI - Ultra-low dose naloxone restores the antinociceptive effect of morphine in pertussis toxin-treated rats by reversing the coupling of mu-opioid receptors from Gs-protein to coupling to Gi-protein. AB - Pertussis toxin (PTX) treatment results in ADP-ribosylation of Gi-protein and thus in disruption of mu-opioid receptor signal transduction and loss of the antinociceptive effect of morphine. We have previously demonstrated that pretreatment with ultra-low dose naloxone preserves the antinociceptive effect of morphine in PTX-treated rats. The present study further examined the effect of ultra-low dose naloxone on mu-opioid receptor signaling in PTX-treated rats and the underlying mechanism. Male Wistar rats implanted with an intrathecal catheter received an intrathecal injection of saline or PTX (1 microg in 5 microl of saline), then, 4 days later, were pretreated by intrathecal injection with either saline or ultra-low dose naloxone (15 ng in 5 microl of saline), followed, 30 min later, by saline or morphine (10 microg in 5 microl of saline). Four days after PTX injection, thermal hyperalgesia was observed, together with increased coupling of excitatory Gs-protein to mu-opioid receptors in the spinal cord. Ultra-low dose naloxone pretreatment preserved the antinociceptive effect of morphine, and this effect was completely blocked by the mu-opioid receptor antagonist CTOP, but not by the kappa-opioid receptor antagonist nor-BNI or the delta-opioid receptor antagonist naltrindole. Moreover, a co-immunoprecipitation study showed that ultra-low dose naloxone restored mu-opioid receptor/Gi-protein coupling and inhibited the PTX-induced mu-opioid receptor/Gs-protein coupling. In addition to the anti-neuroinflammatory effect and glutamate transporter modulation previously observed in PTX-treated rats, the re-establishment of mu opioid receptor Gi/Go-protein coupling is involved in the restoration of the antinociceptive effect of morphine by ultra-low dose naloxone pretreatment by normalizing the balance between the excitatory and inhibitory signaling pathways. These results show that ultra-low dose naloxone preserves the antinociceptive effect of morphine, suppresses spinal neuroinflammation, and reduces PTX-elevated excitatory Gs-coupled opioid receptors in PTX-treated rats. We suggest that ultra low dose naloxone might be clinically valuable in pain management. PMID- 19682559 TI - Sustained elevation of cyclic guanosine monophosphate induces apoptosis in microglia. AB - Cyclic nucleotides mediate transient as well as plastic cellular responses. The most ultimate response is cell death. In the present study, we propose that an increase of intracellular cyclic guanosine monophosphate (cGMP) for at least 1h promotes cell death in the murine microglial cell line, BV-2 cells, as well as in primary murine microglia. Cells were exposed to ammonium, the guanylyl cyclase inhibitor, 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), and to the membrane permeable cGMP analogue, 8-Bromo-cGMP (8-Br-cGMP), respectively. Cell death was estimated using DAPI labelling and annexin-V labelling of exposed phosphatidylserine, and cGMP level was quantified by an immunoassay. Ammonium not only increased the number of apoptotic cells but also promoted a moderate increase in intracellular cGMP. Addition of ODQ suppressed ammonium-induced apoptosis. Furthermore, we found that 8-Br-cGMP significantly increased the number of BV-2 cells and primary microglia, respectively, containing nuclei with condensed chromatin accumulated at the nuclear periphery. Similarly, cells exposed to 8-Br-cGMP showed significantly more cells with exposed phosphatidylserine compared to control cells. Thus, according to the nuclear structure as well as to changes in the plasma membrane, chronic elevation of cGMP induces apoptosis in microglia. PMID- 19682561 TI - "Genomic Impact of Eukaryotic Transposable Elements", Pacific Grove, California, February 6-10, 2009. PMID- 19682560 TI - Author's reply to "cerebral metabolism and sleep homeostasis: a comment on Vyazovskiy et al.". PMID- 19682562 TI - Preparation of rifampicin-loaded PLGA microspheres for lung delivery as aerosol by premix membrane homogenization. AB - The water-in-oil solvent evaporation method with premix membrane homogenization was investigated to improve productivity of the preparation of narrowly size distributed poly(lactide-co-glycolide) (PLGA) microspheres for rifampicin lung delivery as dry aerosols. Using ethyl acetate as organic solvent, a coarse oil-in water emulsion (or premix) was prepared under magnetic stirring and homogenized by extrusion through a Shirasu porous glass (SPG) membrane (5.9 microm porosity). Microspheres were obtained after dilution and solvent evaporation. Formulation parameters investigated were: PLGA concentration, transmembrane pressure and oil:water volume ratio. The optimal formulation parameters were then applied to prepare rifampicin-loaded microspheres. Loaded microspheres were 1.72+/-0.16 microm in diameter with a span of 0.86+/-0.04 and a rifampicin content of 52+/-6 microg/mg microspheres. Release studies in phosphate-buffered saline showed a linear release profile with 40% rifampicin release over 4.5 days. The MMAD of 2.63 microm of freeze-dried microspheres should be suitable for aerosol administration and delivery into the rat lungs by intratracheal insufflation. PMID- 19682563 TI - Influence of protein on mannitol polymorphic form produced during co-spray drying. AB - The stabilizing ability of the excipient on pharmaceutically relevant proteins for potential therapeutic use is an extensive area of research but the effect the protein has on the excipient is rarely reported. The influence of two model proteins on the polymorphic behaviour of mannitol during spray drying was therefore investigated. Spray dried mannitol/protein blends were characterised structurally using X-ray powder diffraction (XRPD) and Fourier transform Raman spectroscopy (FT-Raman) and thermally by differential scanning calorimetry (DSC) and also thermogravimetric analysis (TGA). To assess the long term storage stability, samples were subjected to conditions of elevated temperature and relative humidity (RH). Structural and thermal analysis of the samples showed that upon spray drying mannitol could be completely amorphous or crystalline dependent on the protein co-spray dried. Upon storage at elevated temperature and RH different polymorphic forms of mannitol (beta and delta) were evident again dependent on the protein co-spray dried. Under the conditions employed there was a polymorph directing effect on mannitol dependent on the protein with which it was co-spray dried with co-solute effects on relative water levels being indicated as a major factor in directing the polymorph. PMID- 19682564 TI - Effects of the traditional Japanese medicine Tokaku-jyoki-to in rat-models for menopausal hot flash. AB - AIM OF STUDY: Luteinizing hormone-releasing hormone (LH-RH) has been suggested as an inducer of centrally mediated elevation of skin temperature, and calcitonin gene-related peptide (CGRP) is one of the potent vasodilator neuropeptides that has been suggested as an inducer of peripherally mediated elevation of skin temperature. We investigate the effect of the Japanese herbal medicine Tokaku jyoki-to using two rat-models for menopausal hot flash. MATERIALS AND METHODS: Tokaku-jyoki-to used in present study was prepared as a spray-dried powder from hot-water extract. Skin temperature was measured by thermister thermometer. Estrogen receptor (ER) binding assay of Tokaku-jyoki-to extract was performed using human recombinant ERalpha or ERbeta. RESULTS: Oral Tokaku-jyoki-to (1000 mg/kg) restored skin temperature rise induced by LH-RH or CGRP in ovariectomized (OVX) rats as well as subcutaneous 17beta-estradiol (0.010 mg/kg) did. Tokaku jyoki-to did not affect the lower concentration of plasma estradiol and the decreased uterine weight due to ovariectomy, although the hormone replacement of 17beta-estradiol restored them. In estrogen receptor ligand-binding study, Tokaku jyoki-to extract bound to human ERalpha poorly and did not bound to human ERbeta. CONCLUSIONS: These results suggest that Tokaku-jyoki-to, which appears to contain organ-specific selective estrogen receptor modulator, may be useful for the treatment of hot flashes in patients for whom estrogen replacement therapy is contraindicated as well as menopausal women. PMID- 19682565 TI - Effect of flavonol glycosides from Cinnamomum osmophloeum leaves on adiponectin secretion and phosphorylation of insulin receptor-beta in 3T3-L1 adipocytes. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Cinnamomum osmophloeum is used for various ethnomedical conditions in Taiwan including diabetic complications. AIM OF THE STUDY: The aim of present study was to identify the anti-diabetic compounds from C. osmophloeum leaves and evaluate the preliminary molecular basis for their insulin-like effects. MATERIALS AND METHODS: Silica gel column chromatographic purification of MeOH extract from leaves of C. osmophloeum resulted in the isolation of a two kaempferol glycosides CO-1 and CO-2. These two compounds were evaluated for their effects on adiponectin secretion, tyrosine phosphorylation of insulin receptor (IR)-beta and glucose transporter 4 (GLUT4) in differentiated mouse 3T3-L1 adipocytes, and the results were compared with the reference drug insulin. RESULTS: The compound CO-1 at a concentration of 5 microM was able to act as an insulin-mimetic in terms of its ability to increase adiponectin secretion by 12.2-fold, while CO-2 has no such effect up to 20 microM tested. Furthermore, 5 microM of CO-1 and 20 microM of CO-2 showed potential to increase the phosphorylation of IRbeta by 2.3- and 2.1-fold, respectively, in addition to their positive effect on GLUT4 translocation. CO-1 and CO-2 stimulated GLUT4 translocation are reduced by phosphatidylinositol-3-kinase (PI3-K) inhibitor. CONCLUSION: The present study indicates that the insulin-like anti-diabetic mechanism of constituents from C. osmophloeum leaves in part due to enhanced adiponectin secretion, and activation of insulin signaling pathway leading to GLUT4 translocation which involved phosphorylation of IR and activation of PI3-K. PMID- 19682566 TI - Role of Glutathione monoester on age-related neurochemical alterations in rat brain. AB - It is quite apparent that the incidence of neurodegenerative diseases in both men and women increases in a logarithmic fashion with age and begins to rise much more rapidly after the age of 60. Brain aging is accompanied by structural and functional changes at cellular and tissue levels such as increase in free radical generation, lowered antioxidant defenses, decrease in number of neurons, decrease in the activities of enzymes, (g) decrease in impulse transmission. The present study was aimed to assess the neuromodulatory role of Glutathione monoester (GME) when administered intraperitoneally (12 mg/kg body weight) for 20 days on acetylcholine esterase (AchE) activity, levels of neurotransmitters such as dopamine, serotonin, norepinephrine and rotorod behavioral analysis in discrete brain regions of young and aged male albino Wistar rats. Age-related decrease (p<0.05) in acetylcholine esterase activity, neurotransmitter levels and also decrease in sensorimotor performance was observed. GME administration was effective in restoring these neuronal parameters in aged rat brain regions. Thus GME act as a neuromodulator in discrete brain regions of aged rats. PMID- 19682568 TI - Differential distribution of 4-hydroxynonenal adducts to sulfur and nitrogen residues in blood proteins as revealed using Raney nickel and gas chromatography mass spectrometry. AB - Quantification of 4-hydroxy-2-nonenal (HNE) bound to circulating proteins may prove to be useful in evaluating the role of this bioactive lipoperoxidation by product in the pathogenesis of various diseases. Recently, we developed a quantitative gas chromatography-mass spectrometry (GCMS) assay of total protein bound HNE (HNE-P) in blood after reduction with NaB(2)H(4) and cleavage with Raney nickel. Whereas it has been assumed that Raney nickel cleaves only Michael adducts of HNE to cysteine via a thioether bond (HNE-SP), results from this study demonstrate that our GCMS method also detects with precision picomoles of HNE adducts via nitrogen residues (HNE-NP). Specifically, evidence was obtained using various study models, including polyamino acids consisting of cysteine, lysine, and histidine and a biologically relevant molecule, albumin. Furthermore, we show that dinitrophenylhydrazine treatment before Raney nickel treatment can be used to discriminate and quantify the various HNE-P molecular species in plasma and blood samples from normal rats, which range between 0.15 and 3 pmol/mg protein or 10 to 600 nM. However, whereas HNE-SP predominated in whole blood, we detected HNE-NP only in plasma. We also identified another significant MS signal, which we attribute to protein-bound 1,4-dihydroxynonane (DHN-P) presumably formed from the enzymatic reduction of HNE-P. The distribution profile of all these species in plasma differed from that observed when physiologically relevant concentrations of albumin and HNE were incubated in vitro. Furthermore, interestingly, hypercholesterolemic rabbits showed higher plasma levels of HNE-NP, but not of DHN-P. Beyond documenting the presence of various types of HNE-P in circulating proteins, our results emphasize the importance of enzymatic mechanisms in situ as a factor determining their distribution in the various blood compartments under various conditions. PMID- 19682567 TI - Molecular characterization of phosphorylcholine expression on the lipooligosaccharide of Histophilus somni. AB - Histophilus somni (Haemophilus somnus) is an important pathogen of cattle that is responsible for respiratory disease, septicemia, and systemic diseases such as thrombotic meningoencephalitis, myocarditis, and abortion. A variety of virulence factors have been identified in H. somni, including compositional and antigenic variation of the lipooligosaccharide (LOS). Phosphorylcholine (ChoP) has been identified as one of the components of H. somni LOS that undergoes antigenic variation. In this study, five genes (lic1ABCD(Hs) and glpQ) with homology to genes responsible for ChoP expression in Haemophilus influenzae LOS were identified in the H. somni genome. An H. somni open reading frame (ORF) with homology to H. influenzae lic1A (lic1A(Hi)) contained a variable number of tandem repeats (VNTR). However, whereas the tetranucleotide repeat 5'-CAAT-3' is present in lic1A(Hi), the VNTR in H. somni lic1A (lic1A(Hs)) consisted of 5'-AACC-3'. Due to the propensity of VNTR to vary during replication and cause the ORF to shift in and out of frame with the upstream start codon, the VNTR were deleted from lic1A(Hs) to maintain the gene constitutively on. This construct was cloned into Escherichia coli, and functional enzyme assays confirmed that lic1A(Hs) encoded a choline kinase, and that the VNTR were not required for expression of a functional gene product. Variation in the number of VNTR in lic1A(Hs) correlated with antigenic variation of ChoP expression in H. somni strain 124P. However, antigenic variation of ChoP expression in strain 738 predominately occurred through variable extension/truncation of the LOS outer core. These results indicated that the lic1(Hs) genes controlled expression of ChoP on the LOS, but that in H. somni there are two potential mechanisms that account for antigenic variation of ChoP. PMID- 19682569 TI - Mutational study of the role of N-terminal amino acid residues in tetrachlorohydroquinone reductive dehalogenase from Sphingomonas sp. UG30. AB - This research presents the first extensive mutational study of N-terminal amino acids necessary for activity of a bacterial Zeta class glutathione transferase (GST). Our studies on UG30 tetrachlorohydroquinone reductive dehalogenase (PcpC) revealed that, similar to other Zeta class GSTs, N-terminal Ser and Cys residues play critical roles in glutathione binding and their mutation results in functional and structural changes to PcpC. Mutation of the N-terminal Ser and Cys residues decreased the apparent temperature optimum (by 6-10 degrees C) and maximum (by 5 degrees C) of PcpC. Also, mutation of Ser12 and Ser15 resulted in structural changes that were accompanied by the emergence of substrate inhibition. Mutation of the N-terminal Cys residue adversely affected the rate of the enzymatic reaction, but not on the metabolites formed. This study adds to the knowledge that, despite low sequence homology for the Zeta GST protein family, differences in preferred electrophilic substrates, and the manner in which glutathione is utilized in catalysis, GSTs from prokaryotic and eukaryotic organisms rely upon the same critical amino acids for glutathione binding. PMID- 19682570 TI - Mitochondria in Huntington's disease. AB - Huntington's disease (HD) is an inherited progressive neurodegenerative disorder associated with involuntary abnormal movements (chorea), cognitive deficits and psychiatric disturbances. The disease is caused by an abnormal expansion of a CAG repeat located in exon 1 of the gene encoding the huntingtin protein (Htt) that confers a toxic function to the protein. The most striking neuropathological change in HD is the preferential loss of medium spiny GABAergic neurons in the striatum. The mechanisms underlying striatal vulnerability in HD are unknown, but compelling evidence suggests that mitochondrial defects may play a central role. Here we review recent findings supporting this hypothesis. Studies investigating the toxic effects of mutant Htt in cell culture or animal models reveal mitochondrial changes including reduction of Ca2+ buffering capacity, loss of membrane potential, and decreased expression of oxidative phosphorylation (OXPHOS) enzymes. Striatal neurons may be particularly vulnerable to these defects. One hypothesis is that neurotransmission systems such as dopamine and glutamate exacerbate mitochondrial defects in the striatum. In particular, mitochondrial dysfunction facilitates impaired Ca2+ homeostasis linked to the glutamate receptor-mediated excitotoxicity. Also dopamine receptors modulate mutant Htt toxicity, at least in part through regulation of the expression of mitochondrial complex II. All these observations support the hypothesis that mitochondria, acting as "sensors" of the neurochemical environment, play a central role in striatal degeneration in HD. PMID- 19682571 TI - Regulation of neuron mitochondrial biogenesis and relevance to brain health. AB - Mitochondrial dysfunction has severe cellular consequences, and is linked to aging and neurological disorders in humans. Impaired energy supply or Ca(2+) buffering, increased ROS production, or control of apoptosis by mitochondria may contribute to the progressive decline of long-lived postmitotic cells. Mitochondrial biogenesis refers to the process via which cells increase their individual mitochondrial mass. Mitochondrial biogenesis may represent an attempt by cells to increase their aerobic set point, or an attempt to maintain a pre existing aerobic set point in the face of declining mitochondrial function. Neuronal mitochondrial biogenesis itself has been poorly studied, but investigations from other tissues and model systems suggest a series of transcription factors, transcription co-activators, and signal transduction proteins should function to regulate mitochondrial number and mass within neurons. We review data pertinent to the mitochondrial biogenesis field, and discuss implications for brain aging and neurodegenerative disease research efforts. PMID- 19682572 TI - Cytochrome c oxidase deficiency: patients and animal models. AB - Cytochrome c oxidase (COX) deficiencies are one of the most common defects of the respiratory chain found in mitochondrial diseases. COX is a multimeric inner mitochondrial membrane enzyme formed by subunits encoded by both the nuclear and the mitochondrial genome. COX biosynthesis requires numerous assembly factors that do not form part of the final complex but participate in prosthetic group synthesis and metal delivery in addition to membrane insertion and maturation of COX subunits. Human diseases associated with COX deficiency including encephalomyopathies, Leigh syndrome, hypertrophic cardiomyopathies, and fatal lactic acidosis are caused by mutations in COX subunits or assembly factors. In the last decade, numerous animal models have been created to understand the pathophysiology of COX deficiencies and the function of assembly factors. These animal models, ranging from invertebrates to mammals, in most cases mimic the pathological features of the human diseases. PMID- 19682573 TI - Spatial distribution of the basidiocarps of aphyllophoraceous fungi in a tropical rainforest on Borneo Island, Malaysia. AB - The spatial distribution of basidiocarps provides much information on the dispersal abilities, habitat preferences, and inter- and intraspecific interactions of aphyllophoraceous fungi. To reveal the spatial distribution and resource utilization patterns of aphyllophoraceous fungi in Malaysia, we conducted field observations in a primary forest in 2006 and analyzed the relationships between the abundance of eight dominant fungal species and various environmental factors. The topographical characteristics were significantly patchily distributed at the 100-m scale, whereas woody debris and most fungal species were distributed randomly. Although the dominant fungal species differed among the decay classes and diameters of the woody debris, the abundance of a few dominant species was significantly correlated with environmental factors. Although the latter factors might affect the spatial distribution of these fungi, the effects appear to be so small that they would not create an aggregated distribution at a few 100-m scales. PMID- 19682574 TI - Fungal biomass associated with the phyllosphere of bryophytes and vascular plants. AB - Little is known about the amount of fungal biomass in the phyllosphere of bryophytes compared to higher plants. In this study, fungal biomass associated with the phyllosphere of three bryophytes (Hylocomium splendens, Pleurozium schreberi, Polytrichum commune) and three vascular plants (Avenella flexuosa, Gymnocarpium dryopteris, Vaccinium myrtillus) was investigated using ergosterol content as a proxy for fungal biomass. Phyllosphere fungi accounted for 0.2-4.0 % of the dry mass of moss gametophytes, representing the first estimation of fungal biomass associated with bryophytes. Significantly more fungal biomass was associated with the phyllosphere of bryophytes than co-occurring vascular plants. The ergosterol present in moss gametophytic tissues differed significantly between species, while the ergosterol present in vascular plant leaf tissues did not. The photosynthetic tissues of mosses had less associated fungal biomass than their senescent tissues, and the magnitude of this difference varied in a species specific manner. The fungal biomass associated with the vascular plants studied varied significantly between localities, while that of mosses did not. The observed differences in phyllosphere community biomass suggest their size could be affected by host anatomical and physiological attributes, including micro niche availability and chemical host defenses, in addition to abiotic factors like moisture and nutrient availability. PMID- 19682576 TI - Genetic diversity of Labyrinthula terrestris, a newly emergent plant pathogen, and the discovery of new Labyrinthulid organisms. AB - Labyrinthuloid organisms are thought almost exclusively to be only associated with marine environments. However in 1995, a disease of turfgrass suddenly appeared that was eventually determined to be caused by a new Labyrinthula species (Labyrinthula terrestris). The disease is primarily thought to be caused by the use of elevated salinity irrigation water, making it a unique example of an emergent plant disease potentially induced by human activity. Our objective was to examine diversity of L. terrestris from broadly distributed isolates using AFLP, sequence analysis of two rDNA loci (SSU & LSU-ITS), and pathogenicity tests since previous research on a limited number of isolates found no variability based in ITS and SSU. In contrast to previous work, 18 unique genotypes were found out of a total of 29 analyzed based on AFLP. Sequence variability was only found in a single pathogenic isolate (Laby 31) that was isolated from the United Kingdom. The divergence based on AFLP and sequence analysis suggests that this isolate is a distinct species but closely related to the other L. terrestris isolates examined. Two putatively new nonpathogenic Labyrinthulid species were also found (Laby 13 & 32). Our results suggest that these organisms may be widely distributed in terrestrial environments based on the diversity found in this study and may have long been associated with terrestrial plants. Our results also suggest that more Labyrinthulid organisms may potentially emerge as new plant pathogens in the future if salinification of agricultural systems continues to increases worldwide. PMID- 19682577 TI - Dic2 and Dic3 loci confer osmotic adaptation and fungicidal sensitivity independent of the HOG pathway in Cochliobolus heterostrophus. AB - Previously, we identified three gene loci, Dic1, Dic2, and Dic3, that confer high osmolarity adaptation and dicarboximide/phenylpyrrole fungicide sensitivity in Cochliobolus heterostrophus. Dic1 encoded a group III histidine kinase, but the other genes were not characterized. In the present study, we revealed that both Dic2 and Dic3 are involved in the Skn7 pathway. Dic2 encoded an Skn7-type response regulator, ChSkn7. Strain N4502 contained D359N in the response regulator domain of ChSkn7. Strain E4503 contained a deletion of 50 amino acids in the DNA-binding domain. Strain N4507 was a null mutant of the ChSkn7 gene. All of the dic2 mutant strains showed similar levels of sensitivity to high osmolarity and similar levels of resistance to fungicides. These results strongly suggested that both the DNA-binding domain and response regulator domain are essential for Skn7 function in osmotic adaptation and fungicide sensitivity. A western blot analysis revealed that Dic3 is not involved in the regulation of Hog1-type MAPKs. The Chssk1/dic3 double mutant strains clearly showed greater resistance to fungicides than the single mutant strains. An additive effect was also observed in the high-osmolarity experiments. On the other hand, the dic3/dic2 double mutant strains did not show higher levels of resistance to fungicides and greater sensitivity to KCl than the single mutant strains. These results strongly suggested that the dic3 locus confer high-osmolarity adaptation and fungicide sensitivity independently from Ssk1-Hog1 pathway, but not the Skn7 pathway. Moreover, the dic3 strain and all dic2 strains showed similar levels of sensitivity to high-osmolarity stress and similar levels of resistance to fungicides, suggesting Dic3 to have an essential role in the Skn7 pathway. Our results provide new insight into the functions of the Skn7 pathway in filamentous fungi. PMID- 19682578 TI - Overexpression of Nrp/b (nuclear restrict protein in brain) suppresses the malignant phenotype in the C6/ST1 glioma cell line. AB - Upon searching for glucocorticoid-regulated cDNA sequences associated with the transformed to normal phenotypic reversion of C6/ST1 rat glioma cells, we identified Nrp/b (nuclear restrict protein in brain) as a novel rat gene. Here we report on the identification and functional characterization of the complete sequence encoding the rat NRP/B protein. The cloned cDNA presented a 1767 nucleotides open-reading frame encoding a 589 amino acids residues sequence containing a BTB/POZ (broad complex Tramtrack bric-a-brac/Pox virus and zinc finger) domain in its N-terminal region and kelch motifs in its C-terminal region. Sequence analysis indicates that the rat Nrp/b displays a high level of identity with the equivalent gene orthologs from other organisms. Among rat tissues, Nrp/b expression is more pronounced in brain tissue. We show that overexpression of the Nrp/b cDNA in C6/ST1 cells suppresses anchorage independence in vitro and tumorigenicity in vivo, altering their malignant nature towards a more benign phenotype. Therefore, Nrp/b may be postulated as a novel tumor suppressor gene, with possible relevance for glioblastoma therapy. PMID- 19682579 TI - The effect of murine anti-thymocyte globulin on experimental kidney warm ischemia reperfusion injury in mice. AB - Kidney ischemia-reperfusion injury (IRI) is an important contributor to delayed graft function (DGF) and poor outcome of allografts. Small clinical studies suggest a beneficial role for human anti-thymocyte globulin (ATG) in DGF. We investigated the short-term effect of mouse anti-thymocyte globulin (mATG) on kidney warm IRI in mice. We administered either mATG, rabbit immunoglobulin (RIgG), or saline with different dosing schedules in three different IRI models: 30 min bilateral, 60 min bilateral, and 45min unilateral IRI. mATG effectively depleted circulating T cells but had less effect on kidney-infiltrating T cells. There was no difference in serum creatinine levels between groups in each study. Scoring of renal tubular damage and regenerating tubules revealed no difference between groups. The percentage of CD3(+)CD4(-)CD8(-) double-negative (DN) T cells, which were reported to contribute to the pathogenesis of lupus nephritis, increased and the percentages of regulatory T cells and NK cells decreased in the post-ischemic kidneys of mATG treated mice. mATG did not alter the expression of pro-inflammatory cytokines such as IFN-gamma or anti-inflammatory cytokines such as IL-10 in post-ischemic kidneys. mATG treatment, whether initiated before ischemia or immediately after reperfusion, had minimal effects on renal injury following warm IRI in mice. PMID- 19682581 TI - Role of the PRMT-DDAH-ADMA axis in the regulation of endothelial nitric oxide production. AB - There is abundant evidence that the endothelium plays a crucial role in the maintenance of vascular tone and structure. One of the major endothelium-derived vasoactive mediators is nitric oxide (NO), formed in healthy vascular endothelium from the amino acid precursor l-arginine. Endothelial dysfunction is increased by various cardiovascular risk factors, metabolic diseases, and systemic or local inflammation. One mechanism that has been implicated in the development of endothelial dysfunction is the presence of elevated levels of asymmetric dimethylarginine (ADMA). Free ADMA, which is formed during proteolysis, is actively degraded by the intracellular enzyme dimethylarginine dimethylaminohydrolase (DDAH) which catalyzes the conversion of ADMA to citrulline and dimethylamine. It has been estimated that more than 70% of ADMA is metabolized by DDAH (Achan et al. [1]). Decreased DDAH expression/activity is evident in disease states associated with endothelial dysfunction and is believed to be the mechanism responsible for increased methylarginines and subsequent ADMA mediated eNOS impairment. However, recent studies suggest that DDAH may regulate eNOS activity and endothelial function through both ADMA-dependent and independent mechanisms. In this regard, elevated plasma ADMA may serve as a marker of impaired methylarginine metabolism and the pathology previously attributed to elevated ADMA may be manifested, at least in part, through altered activity of the enzymes involved in ADMA regulation, specifically DDAH and PRMT. PMID- 19682582 TI - Construction and assessment of a 3-T MRI brain template. AB - New MR imaging protocols enable visualization of brain structures. However, for dedicated clinical applications such as targeting deep brain stimulation (DBS), a more accurate localization requires the use of atlases. We developed a three dimensional digitized mono-subject anatomical template of the human brain based on 3-T magnetic resonance images (MRI). By averaging 15 registered T1 image acquisitions, we have shown that the final image corresponds to an optimal image, limited by the performance of the 3-T MR machine. We compared different preprocessing workflows for template construction. With the optimal strategy, along with validated existing processing methods, one T1 template and one T1-T2 mixing template were created in order to improve visualization of spatially complex deep structures. Reduction of voxel size to 0.25 mm(3) was also advantageous to observe fine structures and white matter/gray matter intensity crossings. Results demonstrated that such a template also improved inter-patient registration for population comparison in DBS. These MR templates are made freely available to our community (http://www.vmip.org/mritemplate) to serve as a reference for neuroimage processing methods. PMID- 19682583 TI - Selective impairment of prediction error signaling in human dorsolateral but not ventral striatum in Parkinson's disease patients: evidence from a model-based fMRI study. AB - Animal studies have found that the phasic activity of dopamine neurons during reward-related learning resembles a "prediction error" (PE) signal derived from a class of computational models called reinforcement learning (RL). An apparently similar signal can be measured using fMRI in the human striatum, a primary dopaminergic target. However, the fMRI signal does not measure dopamine per se, and therefore further evidence is needed to determine if these signals are related to each other. Parkinson's disease (PD) involves the neurodegeneration of the dopamine system and is accompanied by deficits in reward-related decision making tasks. In the current study we used a computational RL model to assess striatal error signals in PD patients performing an RL task during fMRI scanning. Results show that error signals were preserved in ventral striatum of PD patients, but impaired in dorsolateral striatum, relative to healthy controls, a pattern reflecting the known selective anatomical degeneration of dopamine nuclei in PD. These findings support the notion that PE signals measured in the human striatum by the BOLD signal may reflect phasic DA activity. These results also provide evidence for a deficiency in PE signaling in the dorsolateral striatum of PD patients that may offer an explanation for their deficits observed in other reward learning tasks. PMID- 19682580 TI - Cellular ADMA: regulation and action. AB - Asymmetric (N(G),N(G)) dimethylarginine (ADMA) is present in plasma and cells. It can inhibit nitric oxide synthase (NOS) that generates nitric oxide (NO) and cationic amino acid transporters (CATs) that supply intracellular NOS with its substrate, l-arginine, from the plasma. Therefore, ADMA and its transport mechanisms are strategically placed to regulate endothelial function. This could have considerable clinical impact since endothelial dysfunction has been detected at the origin of hypertension and chronic kidney disease (CKD) in human subjects and may be a harbinger of large vessel disease and cardiovascular disease (CVD). Indeed, plasma levels of ADMA are increased in many studies of patients at risk for, or with overt CKD or CVD. However, the levels of ADMA measured in plasma of about 0.5micromol.l(-1) may be below those required to inhibit NOS whose substrate, l-arginine, is present in concentrations many fold above the Km for NOS. However, NOS activity may be partially inhibited by cellular ADMA. Therefore, the cellular production of ADMA by protein arginine methyltransferase (PRMT) and protein hydrolysis, its degradation by N(G),N(G)-dimethylarginine dimethylaminohydrolase (DDAH) and its transmembrane transport by CAT that determines intracellular levels of ADMA may also determine the state of activation of NOS. This is the focus of the review. It is concluded that cellular levels of ADMA can be 5- to 20-fold above those in plasma and in a range that could tonically inhibit NOS. The relative importance of PRMT, DDAH and CAT for determining the intracellular NOS substrate:inhibitor ratio (l-arginine:ADMA) may vary according to the pathophysiologic circumstance. An understanding of this important balance requires knowledge of these three processes that regulate the intracellular levels of ADMA and arginine. PMID- 19682584 TI - Dynamic in vivo imaging of cerebral blood flow and blood-brain barrier permeability. AB - The brain is characterized by an extremely rich blood supply, regulated by changes in blood vessel diameter and blood flow, depending on metabolic demands. The blood-brain barrier (BBB)-a functional and structural barrier separating the intravascular and neuropil compartments-characterizes the brain's vascular bed and is essential for normal brain functions. Disruptions to the regional cerebral blood supply, to blood drainage and to BBB properties have been described in most common neurological disorders, but there is a lack of quantitative methods for assessing blood flow dynamics and BBB permeability in small blood vessels under both physiological and pathological conditions. Here, we present a quantitative image analysis approach that allows the characterization of relative changes in the regional cerebral blood flow (rCBF) and BBB properties in small surface cortical vessels. In experiments conducted using the open window technique in rats, a fluorescent tracer was injected into the tail vein, and images of the small vessels at the surface of the cortex were taken using a fast CCD camera. Pixel-based image analysis included registration and characterization of the changes in fluorescent intensity, followed by cluster analysis. This analysis enabled the characterization of rCBF in small arterioles and venules and changes in BBB permeability. The method was implemented successfully under experimental conditions, including increased rCBF induced by neural stimulation, bile salt induced BBB breakdown, and photothrombosis-mediated local ischemia. The new approach may be used to study changes in rCBF, neurovascular coupling and BBB permeability under normal and pathological brain conditions. PMID- 19682585 TI - High-resolution mapping of neuronal activity using the lipophilic thallium chelate complex TlDDC: protocol and validation of the method. AB - In neurons the rate of K(+)-uptake increases with increasing activity. K(+) analogues like the heavy metal ion thallium (Tl(+)) can be used, therefore, as tracers for imaging neuronal activity. However, when water-soluble Tl(+)-salts are injected systemically only minute amounts of the tracer enter the brain and the Tl(+)-uptake patterns are influenced by regional differences in blood-brain barrier (BBB) K(+)-permeability. We here show that the BBB-related limitations in using Tl(+) for imaging neuronal activity are no longer present when the lipophilic Tl(+) chelate complex thallium diethyldithiocarbamate (TlDDC) is applied. We systemically injected rodents with TlDDC and mapped the Tl(+) distribution in the brain using an autometallographic (AMG) technique, a histochemical method for detecting heavy metals. We find that Tl(+)-doses for optimum AMG staining could be substantially reduced, and regional differences attributable to differences in BBB K(+)-permeability were no longer detectable, indicating that TlDDC crosses the BBB. At the cellular level, however, the Tl(+) distribution was essentially the same as after injection of water-soluble Tl(+) salts, indicating Tl(+)-release from TlDDC prior to neuronal or glial uptake. Upon sensory stimulation or intracortical microstimulation neuronal Tl(+)-uptake increased after TlDDC injection, upon muscimol treatment neuronal Tl(+)-uptake decreased. We present a protocol for mapping neuronal activity with cellular resolution, which is based on intravenous TlDDC injections during ongoing activity in unrestrained behaving animals and short stimulation times of 5 min. PMID- 19682586 TI - Gradient distortions in MRI: characterizing and correcting for their effects on SIENA-generated measures of brain volume change. AB - Precise and accurate quantification of whole-brain atrophy based on magnetic resonance imaging (MRI) data is an important goal in understanding the natural progression of neurodegenerative disorders such as Alzheimer's disease and multiple sclerosis. We found that inconsistent MRI positioning of subjects is common in typically acquired clinical trial data - particularly along the magnet's long (i.e., Z) axis. We also found that, if not corrected for, the gradient distortion effects associated with such Z-shifts can significantly decrease the accuracy and precision of MRI-derived measures of whole-brain atrophy - negative effects that increase in magnitude with (i) increases in the Z distance between the brains to be compared and (ii) increases in the Z-distance from magnet isocenter of the center of the pair of brains to be compared. These gradient distortion effects can be reduced by accurate subject positioning, and they can also be corrected post hoc with the use of appropriately-generated gradient-distortion correction fields. We used a novel DUPLO-based phantom to develop a spherical-harmonics-based gradient distortion field that was used to (i) correct for observed Z-shift-associated gradient distortion effects on SIENA generated measures of brain atrophy and (ii) simulate the gradient distortion effects that might be expected with a greater range of Z-shifts than those that we were able to acquire. Our results suggest that consistent alignment to magnet isocenter and/or correcting for the observed effects of gradient distortion should lead to more accurate and precise estimates of brain-related changes and, as a result, to increased statistical power in studies aimed at understanding the natural progression and the effective treatment of neurodegenerative disorders. PMID- 19682587 TI - Improved shimming for fMRI specifically optimizing the local BOLD sensitivity. AB - In functional MRI, magnetic field inhomogeneities due to air-tissue susceptibility differences may lead to severe signal dropouts and geometric distortions in echo-planar images. Therefore, the inhomogeneities in the field are routinely minimized by shimming prior to imaging. However in fMRI, the Blood Oxygen Level Dependent (BOLD) effect is the measure of interest, so the BOLD sensitivity (BS) should be optimized rather than the magnetic field homogeneity. The analytical expression for an estimate of the BOLD sensitivity has been recently developed, allowing for the computation of BOLD sensitivity maps from echo-planar images and field maps. This report describes a novel shimming procedure that optimizes the local BOLD sensitivity over a region of interest. The method is applied in vivo and compared to a standard global shimming procedure. A breath-holding experiment was carried out and demonstrated that the BS-based shimming significantly improved the detection of activation in a target region of interest, the medial orbitofrontal cortex. PMID- 19682588 TI - Study on the neuroprotective effect of fluoxetine against MDMA-induced neurotoxicity on the serotonin transporter in rat brain using micro-PET. AB - 3, 4-Methylenedioxymethamphetamine (MDMA, "ecstasy") has toxic effects on serotonergic neurons in the brain. Our aim was to determine whether N,N-dimethyl 2-(2-amino-4-[(18)F]-fluorophenylthio) benzylamine (4-[(18)F]-ADAM; a serotonin transporter imaging agent) and micropositron emission tomography (micro-PET) can be used to examine in vivo the effect of fluoxetine on MDMA-induced loss of serotonin transporters in rat brain. Male Sprague-Dawley rats were injected with fluoxetine [1 dose, 5 mg/kg, subcutaneously (s.c.)] followed by MDMA (twice a day for 4 consecutive days, 10 mg/kg, s.c.). Micro-PET with 4-[(18)F]-ADAM was performed on days 4, 10, 17, 24, and 31. In addition, the time course of occupancy by fluoxetine at 4-[(18)F]-ADAM sites was measured. Specific 4-[(18)F] ADAM uptake ratios (SURs) were calculated from the micro-PET imaging data for various brain regions. Immunohistochemistry was performed 7 days after the last micro-PET scan. From day 4 to day 31, SURs were markedly decreased (by approximately 55-75% compared to control values) in all brain regions in MDMA treated rats. The effect of MDMA was markedly attenuated (approximately 30-50%) by fluoxetine. The fluoxetine-induced decrease in uptake in different brain regions was 40-75% at 90-min postinjection, and this decrease returned to baseline values in most brain regions by day 31. The distribution and intensity of serotonin transporter (SERT) immunostaining in the brain paralleled the PET imaging results, suggesting that a single dose of fluoxetine provides long lasting protection against MDMA-induced loss of SERT and that such neuroprotection is detectable in vivo by 4-[(18)F]-ADAM micro-PET. PMID- 19682589 TI - Patterns of macroevolution among Primates inferred from a supermatrix of mitochondrial and nuclear DNA. AB - Here, we present a new primate phylogeny inferred from molecular supermatrix analyses of size 42 kb containing 70% of missing data, and representing 75% of primate species diversity. The supermatrix was analysed using a gene-partitioned maximum likelihood approach to obtain an exhaustive molecular phylogenetic framework. All clades recovered from recent molecular works were upheld in our analyses demonstrating that the presence of missing data did not bias our supermatrix inference. The resulting phylogenetic tree was subsequently dated with a molecular dating method to provide a timescale for speciation events. Results obtained from our relaxed molecular clock analyses concurred with previous works based on the same fossil constraints. The resulting dated tree allowed to infer of macroevolutionary processes among the primates. Shifts in diversification rate and speciation rates were determined using the SymmeTREE method and a birthdeath process. No significant asymmetry was detected for the primate clade, but significant shifts in diversification rate were identified for seven clades: Anthropoidea, Lemuriformes, Lemuridae, Galagidae, Callithrix genus, the Cercopithecinae and Asian Macaca. Comparisons with previous primate supertree results reveal that (i) there was a diversification event at the root of the Lemuriformes, (ii) a higher diversification rate is detected for Cercopithecidae and Anthropoidea and (iii) a shift in diversification is always recovered for Macaca genus. Macroevolutionary inferences and primate divergence dates show that major primate diversification events occurred after the Paleogene, suggesting the extinction of ancient primate lineages. PMID- 19682590 TI - A new species of Haemopis (Annelida: Hirudinea): evolution of North American terrestrial leeches. AB - Among the relatively few terrestrial leeches known worldwide, only two (Haemopis terrestris, Haemopis septagon) are described from North America. Here we report a third terrestrial leech collected from the southern part of New Jersey, USA. Tissue samples were obtained from 14 individuals representing three populations, and morphological characters were scored after dorsal and ventral dissections. Maximum Parsimony and Bayesian Inference analyses resolved phylogenetic relationships within the genus Haemopis using cytochrome c oxidase subunit 1 (CO1), 12S ribosomal RNA (rRNA), and 28S rRNA gene fragments, establishing the monophyly of North American haemopids and terrestrialism as a synapomorphy for some members of the group. Morphological distinctions and geographic isolation support the designation of a new species of terrestrial leech, Haemopis ottorum. Phylogeographic interpretations of the haemopid clade suggest that terrestrialism was derived from a northern, aquatic ancestor whose descendents were initially confined to Midwestern States and central Canada by the Appalachian Range. More recently, the terrestrial lineage (i.e., Haemopis terrestris) diverged near the southern extent of its range and began a northeasterly migration along coastal states giving rise to Haemopis septagon and Haemopis ottorum, the latter of which appears to define the leading edge of a northward expansion. PMID- 19682593 TI - Transcriptomics: unravelling the biology of transcription factors and chromatin remodelers during development and differentiation. AB - Mammalian development is a highly complex and tightly regulated process. Transcription factors and chromatin remodelers, acting downstream of cell signalling pathways, are the key intrinsic factors which control gene expression. Recent advances in transcriptomics are allowing biologists to begin to unravel the complex biological roles played by these factors. This review focuses on how genome-wide gene expression and chromatin immunoprecipitation studies are expanding our understanding of the roles played by transcription factors and chromatin remodelers during cell fate decisions in development and differentiation. PMID- 19682594 TI - Live imaging of development in fish embryos. AB - Understanding the cellular and molecular mechanisms that drive the development of embryos requires a detailed knowledge of the way cells divide, move, change shape, interact with one another and die during embryogenesis. Ideally this should be analysed in intact embryos using minimally invasive techniques. Because of their easy accessibility, external development and excellent transparency the teleost embryo has emerged as probably the premier vertebrate model for this type of study. This review will discuss some of the recent advances in this field including attempts to image every cell and their movements during the first 24h of development as well as other studies that focus on the development of specific organs or high resolution analyses of the behaviour of individual cells. PMID- 19682591 TI - Encoding and retrieval are differentially processed by the anterior cingulate and prelimbic cortices: a study based on trace eyeblink conditioning in the rabbit. AB - A growing body of literature suggests that structures along the midline of the prefrontal cortex (mPFC), including Brodmann's area 32 (prelimbic cortex) and area 24 (anterior cingulate cortex) in the rabbit play a role in retrieval of learned information. The present studies compared the effects of post-training lesions produced either immediately or 1-week following learning, to either prelimbic (area 32) or anterior cingulate (area 24) cortex on trace eyeblink (EB) conditioning. Further, because recent evidence suggests that the mPFC may play an even greater role in learning and memory when emotional arousal is low, these studies compared the effects of lesions in groups conditioned with either a relatively low-arousal corneal airpuff, or a more aversive periorbital eyeshock unconditioned stimulus (US). A total of six groups were tested, which received selective ibotenic acid or "sham" control lesions to either area 32 or 24, immediately or 1-week following asymptotic learning, and conditioned with an eyeshock US or an airpuff US. Results showed that the greatest lesion deficits were found when conditioning with the less aversive airpuff US. Further, lesions produced to area 32 one-week, but not immediately following learning, caused significant deficits in performance, while lesions produced to area 24 immediately, but not 1-week following learning, caused significant deficits in performance. These findings add to the body of evidence which shows that area 32 of the mPFC regulates retrieval, but not acquisition or storage of information, while area 24 mediates a less specific reacquisition process, but not permanent storage or retrieval of information during relearning of memories abolished by mPFC damage. These findings were, however, specific to those experiments in which the relatively non-aversive airpuff was the US. PMID- 19682592 TI - Collagen-based cell migration models in vitro and in vivo. AB - Fibrillar collagen is the most abundant extracellular matrix (ECM) constituent which maintains the structure of most interstitial tissues and organs, including skin, gut, and breast. Density and spatial alignments of the three-dimensional (3D) collagen architecture define mechanical tissue properties, i.e. stiffness and porosity, which guide or oppose cell migration and positioning in different contexts, such as morphogenesis, regeneration, immune response, and cancer progression. To reproduce interstitial cell movement in vitro with high in vivo fidelity, 3D collagen lattices are being reconstituted from extracted collagen monomers, resulting in the re-assembly of a fibrillar meshwork of defined porosity and stiffness. With a focus on tumor invasion studies, we here evaluate different in vitro collagen-based cell invasion models, employing either pepsinized or non-pepsinized collagen extracts, and compare their structure to connective tissue in vivo, including mouse dermis and mammary gland, chick chorioallantoic membrane (CAM), and human dermis. Using confocal reflection and two-photon-excited second harmonic generation (SHG) microscopy, we here show that, depending on the collagen source, in vitro models yield homogeneous fibrillar texture with a quite narrow range of pore size variation, whereas all in vivo scaffolds comprise a range from low- to high-density fibrillar networks and heterogeneous pore sizes within the same tissue. Future in-depth comparison of structure and physical properties between 3D ECM-based models in vitro and in vivo are mandatory to better understand the mechanisms and limits of interstitial cell movements in distinct tissue environments. PMID- 19682595 TI - Computational approaches to the integration of gene expression, ChIP-chip and sequence data in the inference of gene regulatory networks. AB - A major challenge in systems biology is the ability to model complex regulatory interactions, such as gene regulatory networks, and a number of computational approaches have been developed over recent years to address this challenge. This paper reviews a number of these approaches, with a focus on probabilistic graphical models and the integration of diverse data sets, such as gene expression and transcription factor binding site location and activity. PMID- 19682596 TI - Nitric oxide and repair of skeletal muscle injury. AB - The muscle wound healing occurs in three overlapping phases: (1) degeneration and inflammation, (2) muscle regeneration, and (3) fibrosis. Simultaneously to injury cellular infiltration by neutrophils and macrophages occur, as well as cellular 'respiratory burst' via activation of the enzyme NADPH oxidase. When skeletal muscle is stretched or injured, myogenic satellite cells are activated to enter the cell cycle, divide, differentiate and fuse with muscle fibers to repair damaged regions and to enhance hypertrophy of muscle fibers. This process depends on nitric oxide (NO) production, metalloproteinase (MMP) activation and release of hepatocyte growth factor (HGF) from the extracellular matrix. Generation of a fibrotic scar tissue, with partial loss of function, can also occur, and seems to be dependent, at least in part, on local TGF-beta expression, which can be downregulated by NO. Hence, regeneration the muscle depends on the type and severity of the injury, the appropriate inflammatory response and on the balance of the processes of remodeling and fibrosis. It appears that in all these phases NO exerts a significant role. Better comprehension of this role, as well as of the participation of other important mediators, may lead to development of new treatment strategies trying to tip the balance in favor of greater regeneration over fibrosis, resulting in better functional recovery. PMID- 19682597 TI - Endothelial nitric oxide synthase is involved in calcium-induced Akt signaling in mouse skeletal muscle. AB - We hypothesized that targeted mutation of the endothelial nitric oxide synthase (eNOS) gene would reduce Akt-related signaling events in skeletal muscle cells, compared to wild type (WT) controls. Results show that slow myosin heavy chain (type I/beta) expression and the abundance of slow-twitch fibers are reduced in plantaris muscle of eNOS(-/-) mice, compared to WT. Further, basal phosphorylation of Akt (p-Akt (Ser-473)/total Akt) and GSK-3beta (GSK-3beta (Ser 9)/total GSK-3beta) are reduced 60-70% in primary myotubes from eNOS(-/-) mice. Treatment with the calcium ionophore, A23187 (0.4 microM, 1 h), increased phosphorylation of Akt and GSK-3beta by approximately 2-fold (P<0.05) in myotubes from WT mice, but had no effect on phosphorylation of these proteins in eNOS(-/-) myotubes. Additionally, A23187 treatment failed to induce nuclear translocation of the transcription factor, NFATc1, in eNOS(-/-) myotubes. Treatment with the nitric oxide donor, propylamine propylamine NONOate (PAPA-NO; 1 microM for 1 h) increased Akt and GSK-3beta phosphorylation, and induced NFATc1 nuclear translocation in WT and eNOS(-/-) myotubes, and eliminated differences from WT in the NOS knockout cultures. Parallel experiments in C2C12 myotubes found that Akt phosphorylation induced by NO or the guanylate cyclase activator, YC-1, is prevented by co-treatment with either a guanylate cyclase or PI3K inhibitor (10 microM ODQ or 25 microM LY2904002, respectively). These data suggest that eNOS activity is necessary for calcium-induced activation of the Akt pathway, and that nitric oxide is sufficient to elevate Akt activity in primary myotubes. NO appears to influence Akt signaling through a cGMP, PI3K-dependent pathway. PMID- 19682599 TI - Use of non-invasive bioluminescent imaging to assess mycobacterial dissemination in mice, treatment with bactericidal drugs and protective immunity. AB - Monitoring the spread of mycobacterium in vivo using biophotonic imaging provides a fast, reliable and sensitive method to evaluate the distribution of the infection. Moreover, this technique allows for a significant reduction in the number of animals required in comparison to conventional anatomopathological studies. Here, we describe for the first time and validate the use of a luciferase-tagged recombinant Mycobacterium bovis BCG for non-invasive bioluminescent imaging of 1) bacterial dissemination in tissues, 2) the efficacy of treatment with anti-mycobacterial drugs and 3) the role of adaptive immune responses in controlling mycobacterial infection in vivo. PMID- 19682600 TI - Cigarette smoking and serum soluble Fas levels: Findings from the JACC study. AB - Cigarette smoking enhances low-grade systemic inflammation in the lung and other organs. Activated immune cells play an important role at early and late stages of inflammation, and in recent years, soluble Fas (sFas), an isoform of death molecule Fas, was found to interfere with the apoptotic pathways of these activated immune cells. The aim of this study was to confirm the association between cigarette smoking and sFas levels in healthy male subjects. We measured serum sFas levels of 4415 male subjects selected as controls for a nested case control study within the large-scale cohort study conducted in Japan, called the JACC Study. Smoking status at baseline was evaluated by a self-administered questionnaire. Least square means of sFas according to smoking status and numbers of cigarettes smoked per day among smokers were calculated and adjusted for possible confounding factors. Mean sFas levels showed an increasing trend across never smokers, past smokers and current smokers, as 2.21 (95% CI: 2.14-2.27) ng/ml, 2.29 (2.22-2.36) ng/ml, and 2.36 (2.30-2.43) ng/ml, respectively. However, no dose-response relationship was observed between the number of cigarettes smoked per day and sFas levels among smokers. PMID- 19682601 TI - Ovary structure in a presocial insect, Elasmucha grisea (Heteroptera, Acanthosomatidae). AB - First generation egg clusters of Elasmucha grisea are more closely guarded than second generation clusters. The ovaries of this species are structured to enhance this behavior. The population of E. grisea from S-W Poland breeds in the spring (May-June) and late summer (July-August). The second generation clutches contain fewer eggs and are destroyed 3-4 days after oviposition by predators and parasitoids. The ovary structure in the studied species differs from that found in other Heteroptera. The average number of ovarioles per ovary is 24 while in the other investigated species the number of ovarioles per ovary is 6-7. Lateral oviducts are elongated and the ovarioles are arranged in a pennate pattern. Each ovariole contains only one growing ovarian follicle. Differentiation of the ovarioles and ovarian follicles is synchronised thus enabling simultaneous oviposition. A comparative analysis of the ovary structure during the life cycle, particularly the presence of atresive ovarian follicles in the ovarioles of egg- and nymph guarding females, as well as the shape and structure of the apical part of the tropharium all support the hypothesis of cooperation between females in E. grisea. A similar ovary structure has been observed in the Coccoidea (Hemiptera, Homoptera) which indicates presocial behavior. PMID- 19682598 TI - Evaluating dual activity LPA receptor pan-antagonist/autotaxin inhibitors as anti cancer agents in vivo using engineered human tumors. AB - Using an in situ cross-linkable hydrogel that mimics the extracellular matrix (ECM), cancer cells were encapsulated and injected in vivo following a "tumor engineering" strategy for orthotopic xenografts. Specifically, we created several three-dimensional (3D) human tumor xenografts and evaluated the tumor response to BrP-LPA, a novel dual function LPA antagonist/ATX inhibitor (LPAa/ATXi). First, we describe the model system and the optimization of semi-synthetic ECM (sECM) compositions and injection parameters for engineered xenografts. Second, we summarize a study to compare angiogenesis inhibition in vivo, comparing BrP-LPA to the kinase inhibitor sunitinib maleate (Sutent). Third, we compare treatment of engineered breast tumors with LPAa/ATXi alone with treatment with Taxol. Fourth, using a re-optimized sECM for non-small cell lung cancer cells, we created reproducibly sized subcutaneous lung tumors and evaluated their response to treatment with LPAa/ATXi. Fifth, we summarize the data on the use of LPAa/ATXi to treat a model for colon cancer metastasis to the liver. Taken together, these improved, more realistic xenografts show considerable utility for evaluating the potential of novel anti-metastatic, anti-proliferative, and anti-angiogenic compounds that modify signal transduction through the LPA signaling pathway. PMID- 19682602 TI - The sensory structures of the antennal flagellum in Hyalesthes obsoletus (Hemiptera: Fulgoromorpha: Cixiidae): a functional reduction? AB - Despite their relevance as harmful pests on plants of economic importance, Hemiptera Fulgoromorpha have been poorly studied as regards their antennal sensory structures. In particular, the flagellum has been neglected and, therefore, to date there are no data on its structural organization and sensory equipment. In order to fill this gap, we carried out a study on the sensillum types and distribution on the flagellum of the planthopper Hyalesthes obsoletus Signoret, an efficient vector of the stolbur phytoplasma, the cause of various crop diseases. In this cixiid species the antenna is composed of three segments, the scape, an enlarged pedicel and a long flagellum. This latter is made of a single segment and presents a basal, bulb-like enlargement from which two processes arise, a short spur and a long arista. Combining scanning electron microscopy, transmission electron microscopy and focused ion beam investigations, we discovered the presence of a total number of 6 sensilla, belonging to 4 different types: a single scolopidium extending from the bulb to the arista, three sensilla styloconica within the cuticular spur and two different sensilla coeloconica inside the bulb. As far as structural data can suggest, these sensilla might be involved in the perception of mechanical stimuli (possibly air borne vibrations), temperature and humidity variations and CO(2) concentration. The strong reduction in sensillum number in this species is discussed as possible functional specialization of the flagellum itself. The ultrastructure of the sensilla in the flagellum of a species of Fulgoromorpha is here presented for the first time. PMID- 19682603 TI - Mitochondrial haplotype and phenotype of 13 Chinese families may suggest multi original evolution of mitochondrial C1494T mutation. AB - Mutations in mitochondrial DNA (mtDNA) are associated with sensorineural hearing loss. In this study, we traced the origin of the 12S rRNA C1494T mutation through analysis of the clinical, genetic, and molecular characteristics of 13 Han Chinese pedigrees with aminoglycoside-induced and non-syndromic bilateral hearing loss that were selected by C1494T screening in 3133 subjects with non-syndromic hearing impairment from 27 regions of China (13/3133). Clinical evaluation revealed the variable phenotypes of hearing impairment including severity, age-of onset, and audiometric configuration in these subjects. Through the whole mitochondrial genome DNA sequence analysis, we identified two evolutionarily conservative variants in protein-coding genes: tRNA(Ala) T 5628C and tRNA(Tyr) A5836G mutations. However, the pedigrees with these mutations did not have a higher or lower penetrance of deafness than in other pedigrees. These results suggested that both T 5628C and A5836G mutations might not significantly modify the manifestation of the C1494T mutation. Sequencing analysis of the whole mitochondrial genome of the probands showed that 13 pedigrees from seven different provinces were classified into 10 haplogroups by the distinct sets of mtDNA polymorphisms, including haplogroups A, B, D, D4, D4b2, F1, M, M7c, N9a1, and H2b. This result suggested that the C1494T mutation occurred sporadically with multi-origins through the evolution of the mtDNA in China, and these mtDNA haplogroup-specific variants may not play an important role in the phenotypic expression of the C1494T mutation in these Chinese families with different penetrance of hearing loss. In addition, the lack of a significant mutation in the GJB2 gene ruled out the possible involvement of GJB2 in the phenotypic expression of the C1494T mutation in those affected subjects. Therefore, the aminoglycosides is solo well-established factor to contribute to the deafness manifestation of the C1494T mutation, and prevention by avoiding the administration of aminoglycosides in individuals carrying C1494T mutation is the most effective way to protect their vulnerability to deafness. PMID- 19682604 TI - Endurance training improves gastrocnemius mitochondrial function despite increased susceptibility to permeability transition. AB - The aim of the present work was to test the hypothesis that moderate endurance treadmill training ameliorates gastrocnemius mitochondrial bioenergetics and increases the tolerance to the calcium-induced mitochondrial permeability transition pore (MPTP) opening. Twelve adult (6-8 week old) male Wistar rats were randomly divided into two groups (n=6per group): sedentary and trained (14 week of endurance treadmill running, 60min/day). Several end-points for invitro gastrocnemius mitochondrial function including oxygen consumption, transmembrane electric potential and susceptibility to calcium-induced MPTP opening were evaluated. Caspase-9 activity was measured in the intact tissue. Endurance training induced significant increases in state 3 and in respiratory control ratio both with complex I and II-linked substrates (malate+pyruvate and succinate, respectively). Increased CCCP-induced uncoupled respiration with succinate as substrate was also observed (p<0.05). No differences were found regarding state 4 and ADP/O ratio with both substrates. In addition, training significantly decreased the phosphorylative lag phase, whereas no changes were observed on maximal transmembrane electric potential, ADP-induced depolarization and repolarization potential (p<0.05). Interestingly and as opposed to our hypothesis, muscle mitochondria isolated from trained rats were more susceptible to MPTP induction by calcium, although in an initial phase muscle mitochondria isolated from trained rats had an increased calcium uptake. Interestingly, we also verified that endurance training increased the activity of caspase 9. The data obtained confirms that endurance training results in a general improvement in the gastrocnemius mitochondrial respiratory function, although mitochondrial and cellular alterations during training also result in increased calcium-induced MPTP opening. PMID- 19682605 TI - Identification of mitochondrial Complex II subunits SDH3 and SDH4 and ATP synthase subunits a and b in Plasmodium spp. AB - While most protist mitochondrial enzymes could be identified in database, the membrane anchor subunits of Complex II and F(o)F(1)-ATP synthase of malaria parasites are not annotated. Based on the presence of structural fingerprints or proteomics data from other protists, here we present their candidates. In contrast to canonical subunits, Plasmodium Complex II anchors have two transmembrane helices and may coordinate heme b via Tyr in place of His. Transmembrane helix IV of ATP synthase subunit a lacks an essential Arg residue. Membrane anchors of Plasmodium Complex II and ATP synthase are divergent from orthologs and promising targets for new chemotherapeutics. PMID- 19682607 TI - Autonomic function following cervical spinal cord injury. AB - Spinal cord injury (SCI) is commonly associated with devastating paralysis. However, this condition also results in a variety of autonomic dysfunctions, primarily: cardiovascular, broncho-pulmonary, urinary, gastrointestinal, sexual, and thermoregulatory. SCI and the resultant unstable autonomic control are responsible for increased mortality from cardiovascular and respiratory disease among individuals with SCI. Injury level and severity directly correlate to the severity of autonomic dysfunctions following SCI. Following high cervical SCI, parasympathetic (vagal) control will remain intact, while the spinal sympathetic circuits will lose their tonic supraspinal autonomic control. On the other hand, in individuals with injury below the 5th thoracic segment, both the sympathetic and parasympathetic control of the heart and broncho-pulmonary tree are intact. As a result of injury level, individuals with quadriplegia versus those with paraplegia will have very different cardiovascular and respiratory responses. Furthermore, similar relationships can exist between the level of SCI and function of other organs that are under autonomic control (bladder, bowel, sweat glands, etc.). It is also important to appreciate that high cervical injuries result in significant respiratory dysfunctions due to the involvement of the diaphragm and a larger portion of the accessory respiratory muscles. Early recognition and timely management of autonomic dysfunctions in individuals with SCI are crucial for the long term health outcomes in this population. PMID- 19682606 TI - The m.3243A>G mtDNA mutation is pathogenic in an in vitro model of the human blood brain barrier. AB - MELAS is a common mitochondrial disease frequently associated with the m.3243A>G point mutation in the tRNA(Leu(UUR)) of mitochondrial DNA and characterized by stroke-like episodes with vasogenic edema and lactic acidosis. The pathogenic mechanism of stroke and brain edema is not known. Alterations in the blood brain barrier (BBB) caused by respiratory chain defects in the cortical microvessels could explain the pathogenesis. To test this hypothesis we developed a tissue culture model of the human BBB. The MELAS mutation was introduced into immortalized brain capillary endothelial cells and astrocytes. Respiratory chain activity and transendothelial electrical resistance, TEER was measured. Severe defects of respiratory chain complex I and IV activities, and a moderate deficiency of complex II activity in cells harboring the MELAS mutation were associated with low TEER, indicating that the integrity of the BBB was compromised. These data support our hypothesis that respiratory chain defects in the components of the BBB cause changes in permeability. PMID- 19682608 TI - Descending bulbospinal pathways and recovery of respiratory motor function following spinal cord injury. AB - The rodent respiratory system is a relevant model for study of the intrinsic post lesion mechanisms of neuronal plasticity and resulting recovery after high cervical spinal cord injury. An unilateral cervical injury (hemisection, lateral section or contusion) interrupts unilaterally bulbospinal respiratory pathways to phrenic motor neurons innervating the diaphragm and leads to important respiratory defects on the injured side. However, the ipsilateral phrenic nerve exhibits a spontaneous and progressive recovery with post-lesion time. Shortly after a lateral injury, this partial recovery depends on the activation of contralateral pathways that cross the spinal midline caudal to the injury. Activation of these crossed phrenic pathways after the injury depends on the integrity of phrenic sensory afferents. These pathways are located principally in the lateral part of the spinal cord and involve 30% of the medullary respiratory neurons. By contrast, in chronic post-lesion conditions, the medial part of the spinal cord becomes sufficient to trigger substantial ipsilateral respiratory drive. Thus, after unilateral cervical spinal cord injury, respiratory reactivation is associated with a time-dependent anatomo-functional reorganization of the bulbospinal respiratory descending pathways, which represents an adaptative strategy for functional compensation. PMID- 19682609 TI - "Short-chain" alpha-1,4-glucan phosphorylase having a truncated N-terminal domain: functional expression and characterization of the enzyme from Sulfolobus solfataricus. AB - All known alpha-1,4-glucan phosphorylases (GPs) are active as homodimers and use their N-terminal domains for oligomerisation. Structure-based sequence comparison of a putative phosphorylase from the thermophilic crenarchaeon Sulfolobus solfataricus (SsGP) with the well characterized GP from Escherichia coli reveals that SsGP totally lacks the otherwise conserved regions for building the dimer interface. Because all efforts of producing functional SsGP in E. coli failed, we used heterologous gene expression in the hyperthermophilic archaeon Thermococcus kodakaraensis and isolated, in low amounts, SsGP harboring Strep-Tag II fused to the C-terminal Tyr-465 of the enzyme. The recombinant protein eluted in size exclusion chromatography with an apparent molecular mass of approximately 69 kDa, consistent with neither the mass expected for a monomer (55 kDa) nor that of a homodimer (110 kDa). The biochemical properties of SsGP were similar to those seen for other GPs containing the N-terminal elements for dimerisation, suggesting that the "short-chain" format of SsGP is fully appropriate for phosphorylase catalytic function and stability. However, the substrate specificity of SsGP differed from that reported for GPs from other thermophilic microorganisms. PMID- 19682610 TI - S1 site residues of Lactococcus lactis prolidase affect substrate specificity and allosteric behaviour. AB - Lactococcus lactis prolidase preferably hydrolyzes Xaa-Pro dipeptides where Xaa is a hydrophobic amino acid. Anionic Glu-Pro and Asp-Pro dipeptides cannot be hydrolyzed at any observable rates and the hydrolysis of cationic Arg-Pro and Lys Pro dipeptides is at about one tenth of the rate of Leu-Pro. It was hypothesized that the hydrophobic residues in the S1 site were responsible for this substrate specificity, thus the residues in the S1 site were substituted with hydrophilic residues. The substitution of Leu193 and Val302 revealed that these residues influenced the substrate specificity. The introduction of a cationic residue, L193R, allowed Asp-Pro to be utilized as a substrate at 37.0% of the rate of Leu Pro, and the anionic mutation, V302D, yielded mutants that could hydrolyze Asp Pro, Arg-Pro and Lys-Pro at 25.9 to 57.4% rates. Interestingly, these mutants of S1 site residues eliminated the allosteric behaviour of L. lactis prolidase that makes this enzyme unique among known prolidases. Results of pH dependency, thermal dependency, and molecular modelling suggested that these observed changes were due to the alteration of the interactions among catalytic zinc cations, Arg293, His296, and the mutated residues. PMID- 19682611 TI - Company profile: Hexascreen Culture Technologies. PMID- 19682613 TI - Chromatin structure regulates human cytomegalovirus gene expression during latency, reactivation and lytic infection. AB - Infection of cells with human cytomegalovirus (HCMV) has two potential outcomes. For instance, infection of fibroblasts results in extensive viral gene expression, viral DNA replication and release of progeny virus. In contrast, in undifferentiated myeloid cells, the lytic transcription programme of HCMV is effectively suppressed and cells undergo latent infection. It is now accepted that the suppression of viral lytic gene expression observed during latency in myeloid cells is a result of the inability of undifferentiated cell types to support robust viral immediate early (IE) gene expression--crucial genes responsible for driving the lytic cycle. The repression of IE gene expression in undifferentiated myeloid cells, at least in part, results from specific post translational modifications of histones associated with the viral major immediate early promoter (MIEP). In cells of the early myeloid lineage, the histone modifications present on the MIEP impart on it a repressive chromatin structure preventing transcriptional activity. Reactivation of HCMV lytic infection is correlated to changes in histone modifications around the MIEP resulting in a chromatin structure conducive to transcriptional activity. These changes are intimately linked with the differentiation of myeloid cells - a phenomenon known to reactivate latent virus in vivo. Chromatin structure of the viral MIEP, therefore, plays a crucial role in latency and reactivation of this persistent human herpesvirus. Whether chromatin-mediated regulation of viral lytic gene expression also occurs, is only beginning to be addressed. However, recent work suggests that all classes of lytic HCMV promoters are subjected to regulation by post-translational modification of their associated histones throughout the time course of infection. Incoming viral genomes appear to be the targets of intrinsic cellular defence mechanisms which attempt to silence viral gene expression through chromatinisation. Viral functions eventually overcome these cellular repression mechanisms permitting high levels of IE gene expression which results in modification of the chromatin structure of early and late gene promoters driving a regulated cascade of viral lytic gene expression and virus production. PMID- 19682615 TI - Association of the herpes simplex virus major tegument structural protein VP22 with chromatin. AB - The abundant tegument protein VP22 has homologs throughout the alphaherpesvirus family. Numerous groups have been involved in the examination of these homologs, trying to decipher their multiple functions. Investigations early on indicated that VP22 associates with chromatin and further evidence has accumulated in recent years, correlating this protein with chromatin and its components. Ongoing and future studies will determine whether this association with chromatin is specific and whether it serves a function during infection in a physiologically relevant setting. PMID- 19682614 TI - Chromatin assembly on herpes simplex virus genomes during lytic infection. AB - The human herpes simplex viruses HSV-1 and HSV-2 infect a significant portion of the human population. Both viruses can undergo lytic infection in epithelial cells and establish lifelong latency in neuronal cells. The large HSV-1 DNA genomes have long been considered to be devoid of histones both inside the virion particle and inside the cell during lytic infection, but to be packaged in repressive chromatin during latency. However, recent reports indicate that many histone and non-histone chromosomal proteins can associate with viral DNA during lytic infection and may influence important events during the HSV-1 lytic cycle. In this article, we summarize recent developments in this field and their implications. PMID- 19682616 TI - Towards an attenuated enterohemorrhagic Escherichia coli O157:H7 vaccine characterized by a deleted ler gene and containing apathogenic Shiga toxins. AB - The aim of this study was to develop a candidate vaccine against enterohemorrhagic Escherichia coli O157:H7. A ler deletion mutant derived from wild-type EHEC O157:H7 86-24 was constructed by use of suicide vector pCVD442. The bacteriophage encoding Shiga toxin (Stx) was excised by serial passage to produce a ler/stx deletion mutant, F25. Stx1 and Stx2 mutants were constructed by site-directed mutagenesis within the active center and membrane-spanning region of the toxin A subunit. Mutants stx1 and stx2 were then introduced into F25 to construct live attenuated candidate vaccine F105. The cytotoxicity of F25 was inactivated and that of F105 was significantly reduced in comparison with wild type E. coli strain EDL933. Mice injected with candidate vaccine strains F25 and F105 gained weight and showed no clinical signs of disease. F25 and F105 reduced the colonization of wild-type O157:H7 in mouse intestine. Immunized pregnant mice were able to protect their suckling newborns from intragastric challenge with wild-type O157:H7. Immunized mice were protected against infection with wild-type O157:H7 and exhibited normal weight gain. Such attenuated vaccine strains may therefore have potential use as oral vaccines against O157:H7. PMID- 19682617 TI - Post-exposure rabies prophylaxis in patients with AIDS. PMID- 19682612 TI - Control of alpha-herpesvirus IE gene expression by HCF-1 coupled chromatin modification activities. AB - The immediate early genes of the alpha-herpesviruses HSV and VZV are transcriptionally regulated by viral and cellular factors in a complex combinatorial manner. Despite this complexity and the apparent redundancy of activators, the expression of the viral IE genes is critically dependent upon the cellular transcriptional coactivator HCF-1. Although the role of HCF-1 had remained elusive, recent studies have demonstrated that the protein is a component of multiple chromatin modification complexes including the Set1/MLL1 histone H3K4 methyltransferases. Studies using model viral promoter-reporter systems as well as analyses of components recruited to the viral genome during the initiation of infection have elucidated the significance of HCF-1 chromatin modification complexes in contributing to the final state of modified histones assembled on the viral IE promoters. Strikingly, the absence of HCF-1 results in the accumulation of nucleosomes bearing repressive marks on the viral IE promoters and silencing of viral gene expression. PMID- 19682618 TI - Cell based vaccination using transplantation of iPSC-derived memory B cells. AB - The recently developed induced pluripotent stem cell (iPSC) technique provides new direction for vaccination: somatic cells can be induced into iPSCs and expanded, then the cells are genetically or chemically promoted to a immune cell fate, followed with in vitro antigen presenting and processing processes to produce memory B cells that can secret functional antibodies to different pathogens; finally these cells are transplanted back to human. PMID- 19682619 TI - Hepatitis B vaccination failure in celiac disease: is there a need to reassess current immunization strategies? AB - Human leukocyte antigen (HLA) phenotype DQ2 is considered the most important genetic marker for un-responsiveness to hepatitis B vaccine. Since celiac disease (CD) is also strongly associated with the same haplo-type it may be hypothesized that celiac patients are less able to respond to the vaccine. We report a retrospective study on celiac patients vaccinated with three doses of 10 microg at 3, 5 and 11 months of age by an intramuscular injection of a recombinant hepatitis B vaccine (Engerix B). We found 30 of 60 celiac patients (50%) unresponsive to vaccination and a significant higher number of responders among patients younger than 18 months at the time of celiac disease diagnosis. Our study confirms that celiac patients have a lower percentage of response to hepatitis B vaccination than healthy subjects. These findings provide useful information to evaluate if current vaccine strategies should be reassessed and if revaccination should be recommended. PMID- 19682620 TI - Re: the power of detention in the management of non-compliance with tuberculosis treatment: a survey of Irish practitioners and analysis of potential legal liability. PMID- 19682622 TI - Radiation therapy for orbital tumors: concepts, current use, and ophthalmic radiation side effects. AB - Radiation therapy is widely used for the treatment of orbital tumors and inflammatory disease. Both external beam teletherapy and implant brachytherapy radiation techniques are employed. External beam radiation therapy is the most common. It involves directing an external radiation source towards the eye, sinuses, and orbit. Whereas most patients are treated with linear accelerator derived external beam radiation therapy, proton, neutron stereotactic radiosurgery, gamma knife, and intensity-modulated radiation therapy have become more available in developed countries. Radiation can be used alone or together with surgery or chemotherapy. Implant radiation therapy (brachytherapy) is also used to treat orbital tumors. Brachytherapy involves surgical placement of radiation sources within the tumor or targeted volume. Characteristically conformal, brachytherapy increases the dose within the target while maximally decreasing exposure of normal tissues. Orbital brachytherapy can be used to boost the dose to the target volume prior to orbital external beam radiation therapy. Herein, I explore the unique challenges associated with irradiation of the orbit, basic radiobiology, doses, indications, and results of treatment. The tolerances of normal ocular and orbital tissues are reported. This review of the literature offers a unique perspective, synthesizing the world's experience on the use of orbital radiation therapy. PMID- 19682621 TI - Intraocular pressure change in orbital disease. AB - Intraocular pressure change has been found concurrent with many orbital pathologies, particularly those involving proptosis. The objective of this review is to offer an inclusive classification of orbital disease-related intraocular pressure change, not only for oculoplastics and glaucoma specialists, but also for general ophthalmologists. Various orbital conditions associated with increased intraocular pressure and glaucoma are comprehensively summarized, and pathophysiology, clinical manifestations, and treatment options of these diseases are discussed. Graves disease, arterio-venous shunts, trauma, and orbital neoplasia, and other common conditions are discussed in detail; less frequent syndromes such as orbitocraniofacial deformities, phakomatoses, and mucopolysaccharidoses are included for the sake of comprehensiveness, but discussed less extensively. PMID- 19682623 TI - Ultrastructural features of posterior crocodile shagreen of the cornea. AB - Three elderly patients with a clinical diagnosis of posterior crocodile shagreen of the cornea underwent penetrating keratoplasty. Each of the patients had the characteristic symmetrical, polygonal opacities with indistinct edges and intervening clear spaces in the central posterior corneal stroma. Electron microscopic examination of the keratoplasty specimens disclosed the presence of vacuoles throughout the corneal stroma, many of which contained electron-dense material. The vacuoles were observed with increasing density posteriorly and were most concentrated adjacent to the anterior banded portion of Descemet's membrane. A sawtooth-like configuration of the stromal collagenous lamellae was noted in the posterior cornea in one case. We reviewed the three cases of central corneal clouding reported with histopathological findings as well as the clinical descriptions of this entity. Although a sawtooth-like lamellar configuration of the stromal collagen may be seen by electron microscopy, characteristic vacuoles appear to be the only consistent finding. We believe that posterior crocodile shagreen is most likely a degenerative disorder and that it should be distinguished from central cloudy dystrophy of Francois by its non-familial pattern of occurrence. PMID- 19682624 TI - The use of vital dyes in ocular surgery. AB - Vital dyes have advanced diagnosis and surgical technique in various specialties, including oncology, gastroenterology, and ophthalmology. In ocular surgery vital dyes are widely used in cataract and vitreoretinal surgery. Worldwide, intra operative use of trypan blue during cataract surgery has enhanced visualization of the anterior capsule during capsulorrhexis, and patent blue has been recently licensed in Europe for cataract surgery. For chromovitrectomy, the vital dyes indocyanine green, infracyanine green, and brilliant blue stain the internal limiting membrane, and trypan blue and triamcinolone acetonide help visualize epiretinal membranes and vitreous, respectively. Intra-operative vital dyes are finding uses in corneal, glaucoma, orbit, strabismus, and conjunctival surgery. We provide a summary of current knowledge of the use of vital dyes in ocular surgery. We review the properties of dyes, techniques of application, indications, and complications in ocular surgery. Vital dyes represent an expanding area of research, and novel dyes deserve further investigation. PMID- 19682626 TI - Bernard Schwartz, MD, PhD, Founding Chair of Ophthalmology, Tufts University School of Medicine, and Founding Editor of Survey of Ophthalmology. AB - Bernard Schwartz, MD, PhD (1927-2007) was first and foremost a scholar. He was a teacher, serving as Professor of ophthalmology for many years at Tufts University; an administrator, chairing the department of ophthalmology for over 20 years; a clinician, practicing primarily in the field of glaucoma; a scientist with a broad interest in ophthalmic research; an historian, especially of medical history; and an editor, primarily of Survey of Ophthalmology. Throughout all of his activities, the central theme was always scholarship. PMID- 19682625 TI - A shot of adrenaline. AB - Acute macular neuroretinopathy is a rare disorder characterized by the sudden onset of unilateral or bilateral paracentral scotomas with relative sparing of the central vision that occurs mostly in young women. It is often characterized by wedge-like macular lesions. The cause of acute macular neuroretinopathy is unknown but viral, immunological, and vascular etiologies have been proposed. There is no current treatment and the visual prognosis is variable. We describe a young woman in whom this disorder was associated with the administration of epinephrine. PMID- 19682627 TI - The enigma of Galileo's eyesight: some novel observations on Galileo Galilei's vision and his progression to blindness. AB - Galileo Galilei became blind. Before this happened he revealed that his left eye had always had less than perfect vision. A study of his written works, his handwriting, and the originals of the portraits undertaken during his lifetime indicate that this probably was the case. These portraits suggest that his left eye tended to lose fixation and that, at the age of 60, he suffered from a mucocoele of the right frontal sinus; but these conditions would not have caused blindness. Considering the systemic diseases from which he suffered over his lifetime, he could possibly have had a long standing uveitis with secondary pupillary block glaucoma, common in those with the group of conditions classified as sero-negative arthropathies. Posterior scleritis with secondary glaucoma is less likely. If either of these were the cause, then the disease was probably triggered by a well-documented, severe acute illness as a young adult, the inflammation being localized to the eye as a result of severe recurrent conjunctival infections in his youth. The intermittent nature of the visual loss, the normal appearance of the cornea and pupils in his portraits, the absence of any evidence of inflammatory joint disease, the presence of halos, and the severe nature of the pain-combined with the high level of visual acuity in between attacks and its persistence until the last few weeks of vision means that angle closure glaucoma must also be considered. These suggestions might be confirmed or refuted by studying his remains. Application has been made for this to be done. PMID- 19682629 TI - Preface: Essays on peripheral nerve repair and regeneration. PMID- 19682628 TI - Mucin 6 in seminal plasma binds DC-SIGN and potently blocks dendritic cell mediated transfer of HIV-1 to CD4(+) T-lymphocytes. AB - Many viruses transmitted via the genital or oral mucosa have the potential to interact with dendritic cell-specific intercellular adhesion molecule-3 grabbing non integrin (DC-SIGN) expressed on immature dendritic cells (iDCs) that lie below the mucosal surface. These cells have been postulated to capture and disseminate human immunodeficiency virus type-1 (HIV-1) to CD4(+) lymphocytes, potentially through breaches in the mucosal lining. We have previously described that BSSL (bile salt-stimulated lipase) in human milk can bind DC-SIGN and block transfer. Here we demonstrate that seminal plasma has similar DC-SIGN blocking properties as BSSL in human milk. Using comparative SDS-PAGE and Western blotting combined with mass spectrometry we identified mucin 6 as the DC-SIGN binding component in seminal plasma. Additionally, we demonstrate that purified mucin 6 binds DC-SIGN and successfully inhibits viral transfer. Mucin 6 in seminal plasma may therefore interfere with the sexual transmission of HIV-1 and other DC-SIGN co-opting viruses. PMID- 19682630 TI - Chapter 1: Peripheral nerve repair and regeneration research: a historical note. AB - Although the most significant advances in nerve repair and regeneration have been acquired over the last few decades, the study of nerve repair and regeneration potential dates back to ancient times namely to Galen in the second century A.D. This brief historical note outlines the milestones which have guided us to our present knowledge. In particular, we focus on the nineteenth century and the first decades of the twentieth century, an age in which the fathers of neurosurgery and neurobiology established the basis for most of the nerve repair and regeneration concepts used today. Finally, we shine a light on the most current history to show how recent pressure to use modern interdisciplinary and translational approach represents a sort of rediscovery of the scientific habits of the fathers of modern biomedicine, who used to carry out research from an integrated and broad point of view rather than from a super-specialized and specific one as it is often used today. PMID- 19682631 TI - Chapter 2: Development of the peripheral nerve. AB - Normal function of the peripheral nerve (PN) is based on morphological integrity and relationship between axons, Schwann cells, and connective sheaths, which depends on the correct development of all these components. Most of the relevant studies in this field were carried out using animal models, since reports on the development of the human PNs from the time of prenatal formation to postnatal development are limited as it is quite difficult to find many nerves in fetuses. In this review paper, we will address the main developmental stages of axons, Schwann cells, and connective tissue sheaths in PNs. Knowledge on the development of PNs and their main components is important for the study of nerve repair and regeneration. This knowledge can be helpful for designing innovative treatment strategies since, like with other organs, the development and regeneration processes share many biological features. PMID- 19682632 TI - Chapter 3: Histology of the peripheral nerve and changes occurring during nerve regeneration. AB - Peripheral nerves are complex organs that can be found throughout the body reaching almost all tissues and organs to provide motor and/or sensory innervation. A parenchyma (the noble component made by the nerve fibers, i.e., axons and Schwann cells) and a stroma (the scaffold made of various connective elements) can be recognized. Although morphological analysis is the most common approach for studying peripheral nerve regeneration, researchers are not always aware of several histological peculiarities of these organs. Therefore, the aim of this review is to describe, at a structural and ultrastructural level, the main features of the parenchyma and the stroma of the normal undamaged nerve as well as the most important morphological changes that occur after nerve damage and during posttraumatic nerve regeneration. The paper is aimed at providing the reader with the basic framework information on nerve morphology. This would enable the correct interpretation of morphological data obtained by many experimental studies on peripheral nerve repair and regeneration such as those outlined in several other papers included in this special issue of the International Review of Neurobiology. PMID- 19682633 TI - Chapter 4: Methods and protocols in peripheral nerve regeneration experimental research: part I-experimental models. AB - This paper addresses several basic issues that are important for the experimental model design to investigate peripheral nerve regeneration. First, the importance of carrying out adequate preliminary in vitro investigation is emphasized in light of the ethical issues and with particular emphasis on the concept of the Three Rs (Replacement, Reduction, and Refinement) for limiting in vivo animal studies. Second, the various options for the selection of the animal species for nerve regeneration research are reviewed. Third, the two main experimental paradigms of nerve lesion (axonotmesis vs. neurotmesis followed by microsurgical reconstruction) are critically outlined and compared. Fourth, the various nerve models that have most commonly been employed are overviewed focusing in particular on forearm mixed nerves and on behavioural tests for assessing their function: the ulnar test and the grasping test which is useful for assessing both median and radial nerves in the rat. Finally, the importance of considering the influence of various factors and diseases which could interfere with the nerve regeneration process is emphasized in the perspective of a wider adoption of experimental models which more closely mimic the environmental and clinical conditions found in patients. PMID- 19682634 TI - Chapter 5: Methods and protocols in peripheral nerve regeneration experimental research: part II-morphological techniques. AB - This paper critically overviews the main procedures used for carrying out morphological analysis of peripheral nerve fibers in light, confocal, and electron microscopy. In particular, this paper emphasizes the importance of osmium tetroxide post-fixation as a useful procedure to be adopted independently from the embedding medium. In order to facilitate the use of any described techniques, all protocols are presented in full details. The pros and cons for each method are critically addressed and practical indications on the different imaging approaches are reported. Moreover, the basic rules of morpho-quantitative stereological analysis of nerve fibers are described addressing the important concepts of design-based sampling and the disector. Finally, a comparison of stereological analysis on myelinated nerve fibers between paraffin- and resin embedded rat radial nerves is reported showing that different embedding procedures might influence the distribution of size parameters. PMID- 19682635 TI - Chapter 6: Methods and protocols in peripheral nerve regeneration experimental research: part III-electrophysiological evaluation. AB - Peripheral nerve injuries usually lead to devastating loss of motor, sensory, and autonomic functions in the patients. Due to the complex requirements for adequate axonal regeneration, functional recovery is often poorly achieved. Experimental models are a useful tool to investigate the mechanisms related to axonal regeneration and reinnervation and to test new strategies to improve functional recovery. Therefore, objective and reliable evaluation methods should be applied for the assessment of regeneration and function restitution after nerve injury in animal models. Electrophysiological tests are commonly used in clinical practice and can be also performed in animal models to determine the nature of peripheral nerve disorders, their severity, and their evolution. These tests provide an integrated approach using sensory and motor nerve conduction studies and electromyography, spinal reflex tests, and motor and sensory evoked potentials if appropriate. The low-invasiveness of several electrophysiological methods allows serial evaluation of sensory and motor reinnervation distal to the injury site at desired intervals without killing the animals or disturbing the regeneration process. This chapter gives a brief review of the most useful electrophysiological methods, their values, and limitations. PMID- 19682636 TI - Chapter 7: Methods and protocols in peripheral nerve regeneration experimental research: part IV-kinematic gait analysis to quantify peripheral nerve regeneration in the rat. AB - Functional recovery is one of the primary goals of therapeutic intervention in peripheral nerve research. The number and diversity of tests which have been used to assess functional recovery after experimental interventions often makes it difficult to recommend any particular indicator of nerve regeneration. Functional assessment after sciatic nerve lesion has long been focused on walking track analysis; however, it is important to note that the validity of the sciatic functional index has been questioned by several researchers. In the last decade, several authors have designed a series of sensitive quantitative methods to assess the recovery of locomotor function using computerized rat gait analysis. The objective of the present review is to provide a helpful tool for the peripheral nerve investigator, by integrating the most important gait kinematic measures described in the literature that can be gathered with this technology. PMID- 19682637 TI - Chapter 8: Current techniques and concepts in peripheral nerve repair. AB - Despite the progress in understanding the pathophysiology of peripheral nervous system injury and regeneration, as well as advancements in microsurgical techniques, peripheral nerve injuries are still a major challenge for reconstructive surgeons. Thorough knowledge of anatomy, pathophysiology, and surgical reconstruction is a prerequisite of proper peripheral nerve injury management. This chapter reviews the currently available surgical treatment options for different types of nerve injuries in clinical conditions. In overview of direct nerve repair, various end-to-end coaptation techniques and the role of end-to-side repair for proximal nerve injuries is described. When primary repair cannot be performed without undue tension, nerve grafting or tubulization techniques are required. Current gold standard for bridging nerve gaps is nerve autografting. However, disadvantages of this approach, such as donor site morbidity and limited length of available graft material encouraged the search for alternative means of nerve gap reconstruction. Nerve allografting was introduced for repair of extensive nerve injuries. Tubulization techniques with natural or artificial conduits are applicable as an alternative for bridging short nerve defects without the morbidities associated with harvesting of autologous nerve grafts. Achieving better outcomes depends both on the advancements in microsurgical techniques and introduction of molecular biology discoveries into clinical practice. The field of peripheral nerve research is dynamically developing and concentrates on more sophisticated approaches tested at the basic science level. Future directions in peripheral nerve reconstruction including, tolerance induction and minimal immunosuppression for nerve allografting, cell based supportive therapies and bioengineering of nerve conduits are also reviewed in this chapter. PMID- 19682638 TI - Chapter 9: Artificial scaffolds for peripheral nerve reconstruction. AB - Posttraumatic peripheral nerve repair is one of the major challenges in restorative medicine and microsurgery. Despite the recent progresses in the field of tissue engineering, functional recovery after severe nerve lesions is generally partial and unsatisfactory. Autograft is still the best method to treat peripheral nerve lesions, although it has several drawbacks and does not allow complete functional recovery. Full recovery of nerve functionality could ideally be achieved by proper guiding axon regeneration toward the original target tissues, through the use of purposely engineered artificial nerve guidance channels (NGCs). In the last decade, artificial NGCs have been produced using a variety of both natural and synthetic, biodegradable and nonbiodegradable polymers. Several techniques have been developed to obtain porous and nonporous NGCs and to realize and incorporate bioactive fillers for NGCs. Some of the developed products have been approved for clinical applications. Many other NGC typologies have been object of interest and are currently under investigation. The current trend of nerve tissue engineering is the realization of biomimetic NGCs, providing chemotactic, topological, and haptotactic signalling to cells, respectively by surface functionalization with cell binding domains, the use of internal-oriented matrices/fibres and the sustained release of neurotrophic factors. The present contribution provides a balanced integration of the most recent achievements of tissue engineering in the field of peripheral nerve repair. By an accurate evaluation of the status of research, the review delineates the most promising directions to which research should address for consistent progress in the field of peripheral nerve repair. PMID- 19682639 TI - Chapter 10: Conduit luminal additives for peripheral nerve repair. AB - The use of nerve conduits as an alternative for nerve grafting has a long experimental and clinical history. Luminal additives, factors introduced into these nerve conduits, were later developed to enhance the nerve regeneration through conduits. This chapter generalizes the types of additives used, and the reported performance of luminal additives in conduits to present a preference list for the most effective additives to use over specific distances of nerve defect. PMID- 19682640 TI - Chapter 11: Tissue engineering of peripheral nerves. AB - Tissue engineering of peripheral nerves has seen an increasing interest over the last years and, similarly to many other fields of regenerative medicine, great expectations have risen within the general public to its potential clinical application in the treatment of damaged nerves. However, in spite of the scientific advancements, applications to the patients is still very limited and it appears that to optimize the strategy for the tissue engineering of the peripheral nerves in the clinical view, researchers have to strive for a new level of innovation which will bring together (in a multitranslational approach) the main pillars of tissue engineering: namely (1) microsurgery, (2) cell and tissue transplantation, (3) material science, and (4) gene transfer. This review paper provides an overview of these four key approaches to peripheral nerve tissue engineering. While some of these issues will also be specifically addressed in other papers in this special issue on peripheral nerve regeneration of the International Review of Neurobiology, in this paper we will focus on an example of successful translational research in tissue engineering, namely nerve reconstruction by muscle-vein-combined nerve scaffolds. PMID- 19682641 TI - Chapter 12: Mechanisms underlying the end-to-side nerve regeneration. AB - End-to-side (ETS) nerve repair is used in selected clinical cases. The mechanisms, by which regeneration into the attached nerve segment is initiated and occur, are still not fully understood. Based on numerous experimental studies, different mechanisms have been suggested by which regenerating axons are recruited, such as contamination from the proximal nerve segment, collateral sprouting, and terminal regenerating sprouting from the donor nerve. A variety of experimental models, most commonly in the lower and upper extremity of rats, and techniques have been used to shed light on the mechanisms. Retrograde labeling techniques have revealed that collateral sprouting do occur, but is probably, at least as observed in long-term experiments, less important over time. Pruning of branching nerve fibers, induced by the collateral sprouting, is an additional mechanism in this context. Experiments have also focused on the stimuli, including the question of epineurial or perineurial windows, that trigger the sprouting of axons form the donor nerve, which can detected by the use of markers of cellular injury. In the present article, we review studies contributing to clarifications of mechanisms of end-to-side nerve repair, including used experimental techniques. We also stress the importance of the plastic brain. PMID- 19682642 TI - Chapter 13: Experimental results in end-to-side neurorrhaphy. AB - Experimental evidence suggests that reinnervation of the distal stump of a transected nerve may occur if the former is coapted end-to-side to the trunk of an adjacent nerve. Axonal regeneration occurs by collateral sprouting of healthy donor nerve axons, induced by neurotrophic factors. End-to-side neurorrhaphy can provide satisfactory functional recovery for the recipient nerve, without any deterioration of the donor nerve function. Various experimental models have been proposed in order to increase regeneration efficiency after end-to-side neurorrhaphy. End-to-side neurorrhaphy has already been used in the clinical practise, but there are still some issues that have not been completely clarified yet: (i) the origin of regenerating axons, (ii) collateral sprouting molecular mechanisms, and (iii) the degree of donor nerve axotomy needed for motor functional recovery. The results of experimental studies trying to investigate these parameters are briefly discussed in this review article. PMID- 19682643 TI - Chapter 14: End-to-side nerve regeneration: from the laboratory bench to clinical applications. AB - Translation of laboratory results to the patient is a critical step in biomedical research and sometimes promising basic science and preclinical results fail to meet the expectations when translated to the clinics. End-to-side (ETS) nerve regeneration is an example of an innovative neurobiological concept, which, after having generated great expectations in experimental and preclinical studies, provided very conflicting results when applied to clinical case series. A number of basic science studies have shown that ETS neurorrhaphy, in fact, is able to induce collateral sprouting from donor nerve's axons, allowing for massive repopulation of the distal nerve stump. Experimental studies have also shown that ETS neurorrhaphy can recover voluntary control of skeletal muscles and that voluntary motor function recovery can be achieved both with agonistic and antagonistic donor nerves, thus widening the potential clinical indications. However, clinical case series reported so far, did not meet these promises and results have been rather conflicting, especially regarding repair of proximally located mixed nerves. In contrast, ETS reconstruction of distal sensory nerve lesions led to a more positive outcome and, most importantly, consistent results among international centers carrying out clinical trials. Concluding, ETS is a promising microsurgical approach for nerve coaptation, based on a convincing and innovative neurobiological concept. However, conflicting clinical results and disagreement among surgeons regarding its employment suggest that this technique should still be considered an ultima ratio, reserved for cases where no other repair technique can be attempted. New data coming from neurobiological research will help further enlarge the clinical indications of ETS nerve reconstruction, explain the different results found in laboratory animals and humans, and contribute to new treatments and rehabilitation strategies aimed at improving the efficacy of nerve regeneration after ETS neurorrhaphy. PMID- 19682644 TI - Chapter 15: Novel pharmacological approaches to Schwann cells as neuroprotective agents for peripheral nerve regeneration. AB - Peripheral neuropathies are common neurologic disorders, but current treatments are limited. Among the different approaches to treat the acquired neuropathies due to traumatic injuries, the pharmacological interventions directed to Schwann cell may represent a useful and challenging opportunity. Following nerve damage the distal axon and the ensheathing Schwann cells degenerate, ensuing a process known as "Wallerian degeneration". Schwann cells then dedifferentiate and proliferate to support neurite outgrowth. In the recent years, several pharmacological agents that may promote the Schwann cell in its role of supporting nerve regeneration have been proposed. However, in view of increased understanding of the cellular mechanisms controlling neuron-glial interactions, a great attention has focused on neurotransmitters, neuroactive steroids, and neurohormones. In this review, we survey the latest findings on these factors and assess their potential as novel promising treatments for peripheral neuropathies caused by injury. PMID- 19682645 TI - Chapter 16: Melatonin and nerve regeneration. AB - Melatonin is a widely distributed and important signal molecule that occurs in unicellular organisms, plants, and fungi in addition to animals and humans. It is the main hormone of the pineal gland and its synthesis occurs mainly in this gland. It has free radical scavenging and antioxidative properties and shows clinical antibacterial and analgesic effects. By means of these properties, it is able to protect cells, tissues, and organs against oxidative damage from free radicals. Recently, widespread interest has grown among researchers regarding the apparent protective effects of melatonin following traumatic events to peripheral nerves, especially the sciatic nerve and its pathological conditions, as melatonin administration could be beneficial following surgery. Although there are great numbers of studies that have mentioned protective effects of melatonin on peripheral nerve pathologies, there are also some studies that report toxic effects of melatonin on peripheral nerves. This paper reviews the available literature in terms of both the beneficial and the toxic effects of melatonin on peripheral nerves. Short descriptions of the structure of pineal gland and synthesis and secretion of melatonin are also given. PMID- 19682646 TI - Chapter 17: Transthyretin: an enhancer of nerve regeneration. AB - Transthyretin (TTR), a plasma and cerebrospinal fluid protein secreted by the liver and choroid plexus, is mainly known as the physiological carrier of thyroxine (T(4)) and retinol. Under pathological conditions, various TTR mutations are related to familial amyloid polyneuropathy (FAP), a neurodegenerative disorder characterized by deposition of TTR amyloid fibrils, particularly in the peripheral nervous system (PNS), leading to axonal loss and neuronal death. Recently, a number of TTR functions in neurobiology have been described; these may explain the preferential TTR deposition, when mutated, in the PNS of FAP patients. In this respect, and with a particular relevance in the PNS, TTR has been shown to have the ability to enhance neurite outgrowth in vitro and nerve regeneration following injury, in vivo. In the following pages, this novel TTR function, as well as its importance in nerve biology and repair will be discussed. PMID- 19682647 TI - Chapter 18: Enhancement of nerve regeneration and recovery by immunosuppressive agents. AB - Clinically, little can be done to induce restoration of good to excellent neurological function following nervous system trauma, and time is required before an effective technique is developed and applied clinically. However, there are novel techniques that have not been tested experimentally or clinically that may induce significantly faster, reliable, and extensive neurological recovery following nervous system trauma than is presently possible, even for techniques currently being tested on animal models. To repair peripheral nerves following trauma in which a length of the nerve pathway is destroyed, many clinicians consider autologous sensory nerve grafts to be the "gold standard" for inducing neurological recovery. However, this technique has severe limitations, such as being effective only across gaps less than 2 cm, for repairs performed less than 2 months posttrauma, and in young patients. As a consequence, many patients suffer permanent neurological deficits or recover only limited neurological function, and they frequently develop irreversible neuropathic pain. This review examines the clinical role that immunosuppressants might play, in the presence or absence of autologous, allografts, or xenografts, in increasing the rate, success, and extent of neurological recovery following nervous system trauma. PMID- 19682648 TI - Chapter 19: The role of collagen in peripheral nerve repair. AB - Collagens are extracellular proteins characterized by a triple helical structure and predominantly involved in the formation of fibrillar and microfibrillar networks of the extracellular matrix and basement membranes. There are 29 collagen types which differ in size, structure, and function. In the peripheral nervous system, two classes of collagen molecules are expressed: fibril forming collagens (type-I, III, and V) and basement membrane collagens (type-IV). Collagens are required for normal extracellular matrix assembly and play an important role in the regulation of Schwann cell function. After injury collagen production in the severed nerve often exceeds the ideal response which is suggested to hinder the growth of sprouting axons into the appropriate distal fascicles and therefore delays and limits nerve regeneration. Both surgical techniques and pharmacological agents are developed to reduce injury induced scarring but despite this nerve regeneration is frequently incomplete. The aim of the present review is to provide the reader a clear overview of the current knowledge with respect to collagens in the peripheral nervous system and to emphasize its role after nerve injury. PMID- 19682650 TI - Chapter 21: Use of stem cells for improving nerve regeneration. AB - A clear need exists for new surgical approaches to enhance the recuperation of functions after peripheral nerve injury and repair. At present, advances in the regenerative medicine fields of biomaterials, cellular engineering, and molecular biology are all contributing to the development of a bioengineered nerve implant, which could be used clinically as an alternative to nerve autograft. In this review we examine the recent progress in this field, looking in particular at the applicability of Schwann cells and stem cell transplantation to enhance nerve regeneration. PMID- 19682649 TI - Chapter 20: Gene therapy perspectives for nerve repair. AB - Despite advances in microsurgical techniques and a good rate of structural nerve healing, functional recovery often remains suboptimal, and thus innovative strategies able to provide extra neurotrophic support to the proper re establishment of functional circuits are warranted. In this review, we will discuss the potential of gene therapy in fostering neuroprotection and neuroregeneration. We will then address a few key emerging concepts in the field, which might help in gaining a better understanding of the cellular and molecular events involved in axonal regeneration, and eventually to the definition of novel targets for intervention. The translation of these new concepts into effective therapies will represent an outstanding challenge for regenerative medicine over the next decades. PMID- 19682651 TI - Chapter 22: Transplantation of olfactory ensheathing cells for peripheral nerve regeneration. AB - Peripheral nerve injury is a common clinical problem, and the development of novel strategies to enhance peripheral nerve regeneration is important. Traumatic events, including motor vehicle accidents, sports-related injuries, violence, and falls, lead to significant numbers of peripheral nerve lesions. Traumatic nerve injuries are often associated with life-threatening injuries, which must be treated first. During the delay in nerve repair, the transected nerves undergo Wallerian degeneration. Therefore, delay before surgical treatment is critical, but care must also be taken to ensure that nerve reapposition is performed in a manner that will result in a therapeutic benefit. Peripheral nerve repair after transection injury combined with transplantation of myelin-forming glia cells, for example, Schwann cells (SCs) or olfactory ensheathing cells (OECs), may facilitate the regenerative process. Cell-based therapies are being considered in clinical trials for a number of neurological diseases, including multiple sclerosis, spinal cord injury, Parkinson's disease, and stroke. The rationale is that transplanted cells may provide neuroprotection by production of chemokines and neurotrophins or could serve as a replacement therapy. A number of cells derived from adult peripheral tissues for cell therapies are also being actively investigated. These cells include SCs from peripheral nerve, olfactory OECs from the olfactory system, and stromal cells from bone marrow (mesenchymal stem cells, MSCs). In principle, these cells could be derived autologously, and used acutely or expanded in culture and used for cell-based therapies. Here, we review experimental work demonstrating the potential of one of these cells, the OEC, as an experimental tool for promoting recovery in peripheral nerve injury. PMID- 19682652 TI - Chapter 23: Manual stimulation of target muscles has different impact on functional recovery after injury of pure motor or mixed nerves. AB - Direct coaptation and interpositional nerve grafting (IPNG) of an injured peripheral nerve is still associated with poor functional recovery. Main reasons for that are thought to be an extensive collateral axonal branching at the site of transection and the polyinnervation of motor endplates due to terminal axonal and intramuscular sprouting. Moreover, severe changes occurring within the muscle after long-term denervation, like loss of muscle bulk and circulation as well as progressive fibrosis, have a negative effect on the quality of functional recovery after reinnervation. We have recently shown that manual stimulation (MS) of paralyzed vibrissal muscles in rat promotes full recovery after facial nerve coaptation. Furthermore, MS improved functional recovery after hypoglossal nerve repair, hypoglossal-facial IPNG of the facial nerve in rat. In contrary, MS did not improve recovery after injury of the median nerve in rat, which is however a mixed peripheral nerve comparing to the facial nerve. It is speculated that manually stimulated recovery of motor function requires an intact sensory input, which is affected in case of mixed peripheral nerves but not in case of pure motor nerves. In this article, we summarize our results of MS in several peripheral nerve injury models in order to illustrate the application potential of this method and to give insights into further investigations on that field. PMID- 19682653 TI - Chapter 24: Electrical stimulation for improving nerve regeneration: where do we stand? AB - While injured neurons regenerate their axons in the peripheral nervous system, it is well recognized that functional recovery is frequently poor. Animal experiments in which injured motoneurons remain without peripheral targets (chronic axotomy) and Schwann cells in distal nerve stumps remain without innervation (chronic denervation) revealed that it is the duration of chronic axotomy and Schwann cell denervation that accounts for this poor functional recovery and not irreversible muscle atrophy that has been so commonly thought to be the reason. More recently, we demonstrated that axon outgrowth across lesion sites is a major contributing factor to the long delays incurred between the injury and the reinnervation of denervated targets. In the rat, a period of 1 month transpires before all motoneurons regenerate their axons across a lesion site. We have developed a technique of 1 h low-frequency electrical stimulation (ES) of the proximal nerve stump just after surgical repair of a transected peripheral nerve that greatly accelerates axon outgrowth. This technique has been applied in patients after carpal tunnel release surgery where the ES promoted the regeneration of all median nerves to reinnervate thenar muscles within 6-8 months, which contrasted with failure of any injured nerves to reinnervate muscles in the same time frame without ES. These findings are very promising such that the ES method could become a clinically viable tool for accelerating axon regeneration and muscle reinnervation. PMID- 19682654 TI - Chapter 25: Phototherapy in peripheral nerve injury: effects on muscle preservation and nerve regeneration. AB - Posttraumatic nerve repair and prevention of muscle atrophy represent a major challenge of restorative medicine. Considerable interest exists in the potential therapeutic value of laser phototherapy for restoring or temporarily preventing denervated muscle atrophy as well as enhancing regeneration of severely injured peripheral nerves. Low-power laser irradiation (laser phototherapy) was applied for treatment of rat denervated muscle in order to estimate biochemical transformation on cellular and tissue levels, as well as on rat sciatic nerve model after crush injury, direct or side-to-end anastomosis, and neurotube reconstruction. Nerve cells' growth and axonal sprouting were investigated in embryonic rat brain cultures. The animal outcome allowed clinical double-blind, placebo-controlled randomized study that measured the effectiveness of 780-nm laser phototherapy on patients suffering from incomplete peripheral nerve injuries for 6 months up to several years. In denervated muscles, animal study suggests that the function of denervated muscles can be partially preserved by temporary prevention of denervation-induced biochemical changes. The function of denervated muscles can be restored, not completely but to a very substantial degree, by laser treatment initiated at the earliest possible stage post injury. In peripheral nerve injury, laser phototherapy has an immediate protective effect. It maintains functional activity of the injured nerve for a long period, decreases scar tissue formation at the injury site, decreases degeneration in corresponding motor neurons of the spinal cord, and significantly increases axonal growth and myelinization. In cell cultures, laser irradiation accelerates migration, nerve cell growth, and fiber sprouting. In a pilot, clinical, double blind, placebo-controlled randomized study in patients with incomplete long-term peripheral nerve injury, 780-nm laser irradiation can progressively improve peripheral nerve function, which leads to significant functional recovery. A 780 nm laser phototherapy temporarily preserves the function of a denervated muscle, and accelerates and enhances axonal growth and regeneration after peripheral nerve injury or reconstructive procedures. Laser activation of nerve cells, their growth, and axonal sprouting can be considered as potential treatment for neural injury. Animal and clinical studies show the promoting action of phototherapy on peripheral nerve regeneration, which makes it possible to suggest that the time for broader clinical trials has come. PMID- 19682655 TI - Chapter 26: Age-related differences in the reinnervation after peripheral nerve injury. AB - Numerous and extensive functional, structural, and biochemical changes characterize intact aged peripheral nervous system. Functional recovery after peripheral nerve injury depends on survival of injured neurons and functional reinnervation of target tissue by regeneration of injured axons and collateral sprouting of uninjured (intact) adjacent axons. The rate of axonal regeneration becomes slower and its extent (density and number of regenerating axons) decreases in aged animals. Aging also impairs terminal sprouting of regenerated axons and collateral sprouting of intact adjacent axons, thus further limiting target reinnervation and its functional recovery. Decreased survival of aged noninjured and injured neurons, limited intrinsic growth potential of neuron, alteration in its responsiveness to stimulatory or inhibitory environmental factors, and changes in the peripheral neural pathways and target tissues are possible reasons for impaired reinnervation after peripheral nerve injury in old age. The review of present data suggests that this impairment is mostly due to the age-related changes in the peripheral neural pathways and target tissues, and not due to the limited intrinsic growth capacity of neurons or their reduced responsiveness to trophic factors. Age-related alterations in the soluble target derived neurotrophic factors, like nerve growth factor, and nonsoluble extracellular matrix components of neural pathways, like laminin, might be important in this respect. PMID- 19682656 TI - Chapter 27: Neural plasticity after nerve injury and regeneration. AB - Injuries to the peripheral nerves result in partial or total loss of motor, sensory, and autonomic functions in the denervated segments of the body due to the interruption of axons, degeneration of distal nerve fibers, and eventual death of axotomized neurons. Functional deficits caused by nerve injuries can be compensated by reinnervation of denervated targets by regenerating injured axons or by collateral branching of undamaged axons, and remodeling of nervous system circuitry related to the lost functions. Plasticity of central connections may compensate functionally for the lack of adequate target reinnervation; however, plasticity has limited effects on disturbed sensory localization or fine motor control after injuries, and may even result in maladaptive changes, such as neuropathic pain and hyperreflexia. After axotomy, neurons shift from a transmitter to a regenerative phenotype, activating molecular pathways that promote neuronal survival and axonal regeneration. Peripheral nerve injuries also induce a cascade of events, at the molecular, cellular, and system levels, initiated by the injury and progressing throughout plastic changes at the spinal cord, brainstem nuclei, thalamus, and brain cortex. Mechanisms involved in these changes include neurochemical changes, functional alterations of excitatory and inhibitory synaptic connections, sprouting of new connections, and reorganization of sensory and motor central maps. An important direction for research is the development of therapeutic strategies that enhance axonal regeneration, promote selective target reinnervation, and are also able to modulate central nervous system reorganization, amplifying positive adaptive changes that improve functional recovery and also reducing undesirable effects. PMID- 19682657 TI - Chapter 28: Future perspective in peripheral nerve reconstruction. AB - Nerve injuries induce severe disability and suffering for patients. Profound alterations in nerve trunks, neurons, and the central nervous system are induced rapidly after injury. This includes activation of intracellular signal transduction mechanisms aiming at the transfer of the cells into a regenerative state through the induction of the appropriate gene programs. The understanding of the neurobiological mechanisms that occur after injury can be used to design modern strategies for reconstruction after nerve injuries. Signal transduction mechanisms for instance may be targets for pharmacological intervention to stimulate nerve regeneration. Nerve injuries, particularly where there is a defect between the severed nerve trunks like in brachial plexus lesions, remain a challenge for the surgeon. Reconstruction of nerve injuries with a defect requires utilization of graft material, which can be of various designs. Application of autologous nerve grafts and use of nerve transfers are the most common clinical solutions to overcome problems with nerve defects. In this chapter we discuss the future perspective of nerve reconstruction with focus on signal transduction mechanisms and new avenues to bridge nerve defects using nanomodified graft surfaces. PMID- 19682658 TI - Livestock waste treatment systems of the future: a challenge to environmental quality, food safety, and sustainability. OECD Workshop. PMID- 19682659 TI - Gender medicine in the American College of Cardiology. PMID- 19682660 TI - Cognitive enhancements in human beings. AB - In the area of genometry-the nascent field of science and technology that proposes to apply enhanced understanding of the human genetic code to reshaping our individual and collective destinies-research and development directed toward improving the function of the human brain has not advanced at the same rate as studies focused on physical enhancement. This article describes developments in the area of cognitive enhancement, including the rise in nonmedical uses of prescription drugs. As those without medical ailments continue to take prescription drugs for their cognitive-enhancing side effects, social policy questions about the fairness of such actions will arise. This article also examines the regulation of cognitive enhancers, both by US law and the rules of international competition. PMID- 19682662 TI - Gender differences in psychotic bipolar mania. AB - OBJECTIVE: This study examined gender differences in the prevalence and types of psychotic symptoms in bipolar mania. METHODS: Participants were drawn from consecutive admissions to the psychiatric clinic in Chemnitz, Germany, in 2005. The diagnosis of bipolar disorder, manic episode was made within 24 hours of admission, and the severity of mania was assessed using the Young Mania Rating Scale (YMRS) and the German version of the Altman Self-Rating Mania Scale. Data collected for each patient included age at the onset of bipolar illness, number of previous episodes, social functioning between episodes, and duration of hospitalization for the index episode. Based on the Task Force for Methods and Documentation in Psychiatry system, psychotic symptoms were classified as hallucinations (visual, auditory, olfactory, tactile, acousma, somatic); delusions (paranoid, reference, guilt, grandeur, religious, erotomania, hypochondriac, poverty, jealousy); and ego disorder (thought control, thought broadcasting). RESULTS: One hundred thirty-seven women and 109 men met the criteria for an acute manic episode, of whom 93 women and 62 men had psychotic symptoms. Compared with psychotic men, psychotic women had more delusions and hallucinations, both overall and per patient, and more delusions of reference and paranoid delusions. Psychotic women had more mixed states compared with psychotic men. Psychotic women differed from both psychotic men and nonpsychotic women on a number of clinical and social variables: they had higher YMRS scores and more previous episodes of depression despite an earlier onset of illness. CONCLUSION: Women with bipolar mania exhibited a specific pattern of psychotic symptoms that appeared to be associated with greater severity of the acute episode, more mixed states, and a more severe course of illness. PMID- 19682661 TI - Effects of gender on response to treatment with rivastigmine in mild cognitive impairment: A post hoc statistical modeling approach. AB - BACKGROUND: Epidemiologic studies have identified several demographic factors, including gender, that may influence the progression from mild cognitive impairment (MCI) to Alzheimer's disease (AD). OBJECTIVE: This analysis aimed to develop a sensitive model for detecting treatment benefits in patients with MCI by controlling for factors that predict progression to AD. The study used this statistical modeling to investigate the effect of gender on treatment response in patients with MCI. METHODS: This post hoc analysis used data from the InDDEx (Investigation in Delay to Diagnosis of Alzheimer's disease with Exelon) study, a long-term (3- to 4-year), multicenter, randomized, double-blind, placebo controlled trial of rivastigmine 3- to 12-mg/d capsules in 1018 patients with MCI. Baseline variables that were significantly associated with progression to AD within the InDDEx study population were identified. A Cox proportional hazards multivariate regression model that adjusted for these predictive factors was applied to the overall study population and to the gender subgroups. RESULTS: Of 31 baseline variables analyzed, 13 were found to be significantly associated with progression to AD in the overall population. After adjustment using the Cox proportional hazards regression model, rivastigmine was associated with a significantly lower risk for progression to AD compared with placebo in the overall population (1016 patients; hazard ratio [HR] = 0.747; P = 0.045). This effect of rivastigmine was evident in women (530 patients; HR = 0.676; P = 0.046) but not in men. CONCLUSIONS: In these patients with MCI, a significant decrease in the risk for progression to AD was found with rivastigmine over 3 to 4 years. The proportion of women whose disease progressed to AD was significantly lower in the rivastigmine group than the placebo group (P = 0.046). This effect was not found in men. These data suggest that prospectively adjusting for predictive factors could lead to a more accurate estimation of treatment benefits in future studies of MCI. PMID- 19682663 TI - Gender differences in self-reported use, perceived efficacy, and interest in future use of nicotine-dependence treatments: A cross-sectional survey in adults at a tertiary care center for nicotine dependence. AB - BACKGROUND: Clinical trials have assessed the effectiveness of nicotine dependence treatments (NDTs), alone or in combination, and reported that men and women have variable responses to these treatments. The variations in therapeutic responses highlight the need to explore gender-specific preferences for NDTs, including complementary and alternative medicine (CAM), which has become increasingly popular in the US population for the cessation of tobacco use. OBJECTIVE: The aim of this study was to assess gender differences in the self reported use, perceived efficacy, and interest in future use of NDTs, including CAM, in an outpatient setting. METHODS: This cross-sectional survey was conducted in men and women at a tertiary care NDT clinic. The primary inclusion criterion was the willingness and ability of the patients to participate in the survey. RESULTS: Data from 1171 patients were included (599 men, 572 women; mean age: men, 46.2 years; women, 46.5 years). Of these, 68% of women and 65% of men reported use of nicotine-replacement therapy (NRT), other prescription medication, or counseling/group support. In men and women, NRT was the most commonly used type of pharmacologic treatment, of which the patch was the most popular (77% and 75%). A significantly greater proportion of women than men perceived the nicotine inhaler to be efficacious (67% vs 50%; P = 0.027). No other significant gender differences were found among NRTs. Among non-NRT methods, bupropion sustained release (SR) and counseling/group support were used by significantly more women than men (53% vs 43% [P = 0.007] and 16% vs 11% [P = 0.026], respectively). Compared with men, significantly greater proportions of women reported current or previous use of CAM for nicotine abstinence and expressed an interest in future use of CAM (34% vs 22% [P < 0.001] and 71% vs 64% [P = 0.006]). CONCLUSIONS: In this sample of patients at an NDT clinic, significantly more women than men reported previous use of bupropion SR, counseling, and CAM. More women than men expressed an interest in the future use of CAM. Based on these findings, an improved understanding of gender-based differences in the use of conventional and nonconventional NDTs might improve the rates of success of nicotine-cessation efforts among women. PMID- 19682664 TI - Sex differences in body composition early in life. AB - BACKGROUND: Early development of the percentage of fat and muscle is rarely considered, but is important because excessive fat is related to the development of diabetes and other morbidities later in life. In pediatric medicine, there are few to no data comparing sex differences in body composition in the first months of life despite the fact that males are typically longer and weigh more than girls at birth. OBJECTIVE: The purpose of this study was to determine whether observed sex differences in body composition at birth persist through the first 6 months of life. METHODS: Participants were healthy, full-term, male and female newborns. Children throughout the Oklahoma City, Oklahoma, metropolitan area were enrolled. The inclusion criteria were: mothers aged 18 to 45 years at the time of delivery; a term pregnancy lasting >or=37 weeks of gestation (determined by mother's physician); weight adequate for gestational age; and a hospital stay for the infant of <3 days following delivery. The exclusion criteria were: maternal tobacco use or alcohol consumption (>1 drink per week) during pregnancy; gestational diabetes; preeclampsia; and infants with presumed or known congenital birth defects. Baseline assessment at birth included length and weight. Newborns had their body composition (percent fat [%fat], total fat, and fat-free mass) determined at approximately 1 month of age using whole body plethysmography. Mothers were invited to have their children take part in a 5-month extension that conducted additional body composition measurements at 3 and 6 months of age. RESULTS: Sixty-four girls (mean [SD] age at time of testing, 20.9 [7.9] days; birth weight, 3500 [388] g; birth length, 49.9 [2.4] cm; white race, 73.4%) and 53 boys (mean age at time of testing, 20.2 [7.3] days; birth weight, 3353 [413] g; birth length, 51.0 [2.4] cm; white race, 69.8%) were assessed and included in the study. At birth, girls were significantly shorter and weighed more than boys (both, P < 0.05). At ~1 month of age, body composition revealed that girls had significantly greater %fat (15.1% vs 12.7%; P < 0.05) and less fat-free mass (3182 [303] vs 3454 [361] g; P < 0.001) than did boys. At 3 months of age, girls continued to have significantly less fat-free mass (4379 [347] vs 4787 [310] g; P < 0.01) than did boys; however, by 6 months of age, no significant sex difference was observed in any body composition variable studied. CONCLUSION: In this small sample of healthy, full-term newborns, at ~1 month of age, statistically significant differences in %fat and fat-free mass existed between girls and boys; however, by 6 months of age, these differences no longer existed. PMID- 19682665 TI - Sex differences in intraventricular hemorrhage rates among very low birth weight newborns. AB - BACKGROUND: The influence of male or female sex on newborn outcomes has been recognized for >30 years. Several studies have observed higher mortality and morbidity in males than in females. It is not clear how this sex difference is sustained in postnatal complications such as intraventricular hemorrhage (IVH), especially in very low birth weight (VLBW) newborns. OBJECTIVE: This study examined possible sex-related differences in IVH rates among VLBW neonates. METHODS: In a retrospective observational study conducted in Hospital Privado, Cordoba, Argentina, data from 332 consecutive VLBW newborns in a 12-year period were reviewed. Maternal factors, labor and delivery characteristics, and neonatal parameters, including the results of cranial ultrasound examination to detect IVH, were compared for males and females. Bivariate and multivariate logistic regression analyses were performed. RESULTS: A total of 322 VLBW newborns were included, 168 males and 154 females. Compared with female neonates, male neonates had a higher risk of overall IVH (26.8% vs 9.7%; odds ratio [OR] = 3.4 [95% CI, 1.8-6.4]; P < 0.001) and for grades III or IV on the Papile scale (16.1% vs 1.9%; OR = 9.6 [95% CI, 2.9-32.5]; P < 0.001). In the multivariate logistic regression model, male sex sustained the association with a greater risk of IVH (OR = 6.8 [95% CI, 3.8-12.0]). CONCLUSIONS: IVH was significantly associated with male sex in these VLBW newborns. Because other factors affect these differences, further research is required. PMID- 19682666 TI - Complementary medicine in the primary care setting: Results of a survey of gender and cultural patterns in Israel. AB - OBJECTIVE: The purpose of this study was to examine the use of complementary and alternative medicine (CAM) in a primary care practice in Israel to determine prevalence and patterns of use. METHODS: Trained research assistants invited all patients attending the administrative, medical, pharmaceutical, or nursing services of 7 clinics in urban and rural areas of northern Israel over a 16-month period, from April 1, 2005, through August 1, 2006, to complete a 13-item written questionnaire about CAM use and beliefs about CAM safety and efficacy. CAM was defined as therapies often referred to as alternative, complementary, natural, or folk/traditional medicine, and which are not usually offered as part of the medical treatment in the clinic, including herbal medicine, Chinese medicine (including acupuncture), homeopathy, folk and traditional remedies, dietary/nutritional therapy (including nutritional supplements), chiropractic, movement/manual healing therapies (including massage, reflexology, yoga, and Alexander and Feldenkrais techniques), mind-body techniques (including meditation, guided imagery, and relaxation), energy and healing therapies, and other naturopathic therapies. The Pearson chi(2) test and multivariate logistic regression were used to assess univariate associations with the odds ratios of CAM use among Arab and Jewish women. A t test was performed to determine whether there were any differences in the continuous variables between the 2 groups. RESULTS: Of 3972 consecutive patients who received the questionnaire, 3447 responded; 2139 respondents (62%) were women. Of the female respondents, 2121 reported their religion (1238 respondents [58%] self-identified as being Arab, and 883 [41.6%] as being Jewish). Compared with men, more women used CAM during the previous year (46.4% vs 39.4%; P < 0.001). Women were more likely to use CAM and to be interested in receiving CAM at primary care clinics. Arab women reported less CAM use than Jewish women but were more interested in experiencing CAM, had a higher degree of confidence in CAM efficacy and safety, and more frequently supported the integration of CAM practitioners in primary care clinics. CONCLUSIONS: In this study, women visiting primary care clinics in northern Israel used CAM more often than men did. Arab women reported less use of CAM than did Jewish women but also reported greater confidence in CAM efficacy and safety. PMID- 19682667 TI - The obesity paradox, weight loss, and coronary disease. AB - PURPOSE: Because obesity is a cardiovascular risk factor but is associated with a more favorable prognosis among cohorts of cardiac patients, we assessed this "obesity paradox" in overweight and obese patients with coronary heart disease enrolled in a cardiac rehabilitation and exercise training (CRET) program, making this assessment in patients classified as overweight/obese using both traditional body mass index (BMI) and percent body fat assessments. Additionally, we assessed the efficacy and safety of purposeful weight loss in overweight and obese coronary patients. PATIENTS AND METHODS: We retrospectively studied 529 consecutive CRET patients following major coronary events before and after CRET, and compared baseline and post program data in 393 overweight and obese patients (body mass index [BMI] > or =25 kg/m(2)) divided by median weight change (median= 1.5%; mean +2% vs -5%, respectively). In addition, we assessed 3-year total mortality in various baseline BMI categories as well as compared mortality in those with high baseline percent fat (>25% in men and >35% in women) versus those with low baseline fat. RESULTS: Following CRET, the overweight and obese with greater weight loss had improvements in BMI (-5%; P <.0001), percent fat (-8%; P <.0001), peak oxygen consumption (+16%; P <.0001), low-density lipoprotein cholesterol (-5%; P <.02), high-density lipoprotein cholesterol (+10%; P <.0001), triglycerides (-17%; P <.0001), C-reactive protein (-40%; P <.0001), and fasting glucose (-4%; P=.02), as well as marked improvements in behavioral factors and quality-of-life scores. Those with lower weight loss had no significant improvements in percent fat, low-density lipoprotein cholesterol, triglycerides, C-reactive protein, and fasting glucose. During 3-year follow-up, overall mortality trended only slightly lower in those with baseline overweightness/obesity who had more weight loss (3.1% vs 5.1%; P=.30). However, total mortality was considerably lower in the baseline overweight/obese (BMI > or =25 kg/m(2)) than in 136 CRET patients with baseline BMI <25 kg/m(2) (4.1% vs 13.2%; P <.001), as well as in those with high baseline fat compared with those with low fat (3.8% vs 10.6%; P <.01). CONCLUSIONS: Purposeful weight loss with CRET in overweight/obese coronary patients is associated with only a nonsignificant trend for lower mortality but is characterized by marked improvements in obesity indices, exercise capacity, plasma lipids, and inflammation, as well as behavioral factors and quality of life. Although an "obesity paradox" exists using either baseline BMI or baseline percent fat criteria, these results support the safety and potential long-term benefits of purposeful weight loss in overweight and obese patients with coronary heart disease. PMID- 19682669 TI - Reducing psychosocial stress: a novel mechanism of improving survival from exercise training. AB - BACKGROUND: Exercise training reduces mortality in patients with coronary artery disease. Behavioral characteristics, including depression, hostility, and overall psychosocial stress, have been shown to be independent risk factors for recurrent myocardial infarction and death in these patients. Exercise training can reduce these high-risk behaviors, but it remains uncertain as to what extent the health benefits of exercise training can be attributed to improving these behaviors. METHODS: We evaluated the impact of exercise training during cardiac rehabilitation on mortality in 53 patients with coronary artery disease with high levels of psychosocial stress and in 469 patients with coronary artery disease with low levels of psychosocial stress and compared them with 27 control patients with high psychosocial stress who did not undergo formal cardiac rehabilitation and exercise training. RESULTS: Mortality was approximately 4-fold greater in patients with high psychosocial stress than in those with low psychosocial stress (22% vs 5%; P = .003). Exercise training decreased the prevalence of psychosocial stress from 10% to 4% (P<.0001) and similarly improved peak oxygen uptake in patients with high and low psychosocial stress. Mortality in patients who improved exercise capacity by>or=10% (high exercise change) was 60% lower than in patients who had<10% improvement in exercise capacity (low exercise change) (P=.009). Mortality was lower in patients with high psychosocial stress with high exercise change compared with patients with high psychosocial stress with low exercise change (0% vs 19%; P=.009). In contrast, there was no significant improvement in mortality in patients with high versus low exercise change with low psychosocial stress (4% vs 8%; P=.14). CONCLUSION: Psychosocial stress is an independent risk factor for mortality in patients with coronary artery disease, and exercise training can effectively reduce its prevalence. Exercise training reduces mortality in patients with coronary artery disease, and this effect seems to be mediated in part because of the salutary effects of exercise on psychosocial stress. PMID- 19682668 TI - Inflammation and thrombosis biomarkers and incident frailty in postmenopausal women. AB - BACKGROUND: The immune and blood coagulation systems have been implicated in the pathophysiology of the geriatric syndrome of frailty, but limited prospective data examining the relationship of clotting/inflammation biomarkers to risk of incident frailty exist. METHODS: This prospective analysis was derived from a nested case-control study within the Women's Health Initiative. Among women 65 to 79 years free of frailty at enrollment, we randomly selected 900 incident cases from those developing frailty within 3 years; 900 non-frail controls were individually matched on age, ethnicity, and blood collection date. Biomarkers assessed for risk of incident frailty included fibrinogen, factor VIII, D-dimer, C-reactive protein, interleukin-6, and tissue plasminogen activator (t-PA). RESULTS: When examined by quartiles in multivariable adjusted models, higher D dimer and t-PA levels were each associated with increased risk of frailty (P trend = .04). Relative to the lowest quartile, the odds ratios for frailty compared with the upper quartile were 1.52 (95% confidence interval, 1.05-2.22) for t-PA and 1.57 (95% confidence interval, 1.11-2.22) for D-dimer. For women having high t-PA and high D-dimer compared with women having lower levels of both biomarkers, the odds of frailty was 2.20 (1.29-3.75). There was little evidence for association between coagulation factor VIII, fibrinogen, C-reactive protein, or interleukin-6 levels and incident frailty. CONCLUSION: This prospective analysis supports the role of markers of fibrin turnover and fibrinolysis as independent predictors of incident frailty in postmenopausal women. PMID- 19682670 TI - Incidence of thrombocytopenia in hospitalized patients with venous thromboembolism. AB - PURPOSE: To determine the incidence of heparin-associated thrombocytopenia in patients receiving prophylaxis or treatment for venous thromboembolism. METHODS: We assessed the database of the National Hospital Discharge Survey from 1979 through 2005 and complemented this with a meta-analysis of published literature. RESULT: Among 10,554,000 patients discharged from short-stay hospitals throughout the US with venous thromboembolism during the 27 years of study, secondary thrombocytopenia was coded in 38,000 patients (0.36%). From 1979 through 1992, secondary thrombocytopenia was coded in only 0.15% of hospitalized patients with venous thromboembolism. The frequency increased sharply to 0.54% from 1993 through 2005. Secondary thrombocytopenia was rarely diagnosed among 1,446,000 patients aged <40 years and among 77,000 women who had venous thromboembolism with deliveries. Meta-analysis of published literature showed a higher incidence among patients who received unfractionated heparin (UFH) for prophylaxis than those who received low-molecular-weight heparin (LMWH) for prophylaxis. Treatment resulted in smaller differences of the incidence between UFH and LMWH. CONCLUSION: Heparin-associated thrombocytopenia is rare among patients aged <40 years and women following delivery. The risk of heparin-associated thrombocytopenia is more duration-related than dose-related, and higher with UFH when used for an extended duration. Our findings and those of the literature suggest that although heparin-associated thrombocytopenia is uncommon, the incidence can be minimized by use of LMWH, particularly if extended prophylaxis or extended treatment is required. PMID- 19682671 TI - Negative self-evaluations and the relation to performance level in socially anxious children and adolescents. AB - The current study investigated whether negatively biased self-evaluations of nervousness and social skills are related to how well an individual actually performs, that is performance level. Sixty-eight high socially anxious and 68 control participants (age range 9-17 years) gave a 5 min speech in front of a pre recorded audience of same age peers and a teacher. Participants' evaluations immediately after the task were measured on a number of performance dimensions. Three independent observers also evaluated recordings of the speech performances. Participants were further divided into good and bad performers based on their actual performance level as judged by the observers. Self-evaluations of the high socially anxious participants were negatively biased for nervous appearance regardless of how well they actually performed. In contrast, a negative bias for social skills only occurred in the high anxious participants with a good performance. The social skill evaluations of the poor performers appear warranted. Taking actual performance level into account may help to clarify the exact nature of a negative bias in socially anxious youth and has clear implications for the choice of treatment approach. PMID- 19682672 TI - Embodied communication: speakers' gestures affect listeners' actions. AB - We explored how speakers and listeners use hand gestures as a source of perceptual-motor information during naturalistic communication. After solving the Tower of Hanoi task either with real objects or on a computer, speakers explained the task to listeners. Speakers' hand gestures, but not their speech, reflected properties of the particular objects and the actions that they had previously used to solve the task. Speakers who solved the problem with real objects used more grasping handshapes and produced more curved trajectories during the explanation. Listeners who observed explanations from speakers who had previously solved the problem with real objects subsequently treated computer objects more like real objects; their mouse trajectories revealed that they lifted the objects in conjunction with moving them sideways, and this behavior was related to the particular gestures that were observed. These findings demonstrate that hand gestures are a reliable source of perceptual-motor information during human communication. PMID- 19682673 TI - Visual perspective taking impairment in children with autistic spectrum disorder. AB - Evidence from typical development and neuroimaging studies suggests that level 2 visual perspective taking - the knowledge that different people may see the same thing differently at the same time - is a mentalising task. Thus, we would expect children with autism, who fail typical mentalising tasks like false belief, to perform poorly on level 2 visual perspective taking as well. However, prior data on this issue are inconclusive. We re-examined this question, testing a group of 23 young autistic children, aged around 8years with a verbal mental age of around 4years and three groups of typical children (n=60) ranging in age from 4 to 8years on a level 2 visual perspective task and a closely matched mental rotation task. The results demonstrate that autistic children have difficulty with visual perspective taking compared to a task requiring mental rotation, relative to typical children. Furthermore, performance on the level 2 visual perspective taking task correlated with theory of mind performance. These findings resolve discrepancies in previous studies of visual perspective taking in autism, and demonstrate that level 2 visual perspective taking is a mentalising task. PMID- 19682674 TI - Object representations maintain attentional control settings across space and time. AB - Previous research has revealed that we create and maintain mental representations for perceived objects on the basis of their spatiotemporal continuity. An important question is what type of information can be maintained within these so called object files. We provide evidence that object files retain specific attentional control settings for items presented inside the object, even when it disappears from vision. The objects were entire visual search displays consisting of multiple items moving into and out of view. It was demonstrated that search was speeded when the search target position was repeated from trial to trial, but especially so when spatiotemporal continuity suggested that the entire display was the same object. We conclude that complete spatial attentional biases can be stored in an object file. PMID- 19682676 TI - Language dominance, handedness and sex: recessive X-linkage theory and test. AB - The possibility is investigated that cerebral dominance for language and handedness share a common X-linkage and that the relation between the two is therefore a function of sex. In particular, an X-linked recessive account is shown to predict an overall configuration of language dominance, handedness and sex within which there is a sex effect in the pattern of language dominance among right-handed but not left-handed people. The recent accurate determination of cerebral dominance among relatively large samples of the general population by means of functional transcranial Doppler ultrasonography makes it possible to test this new theory rigorously, and its parameter-free pattern of predictions is found to be supported. PMID- 19682677 TI - Dose-response relationships of rat fetal skeleton variations: Relevance for risk assessment. AB - In developmental toxicity studies, skeleton abnormalities found in fetuses at term are classified as variations or malformations. The relevance of skeleton variations for human risk assessment, however, is a controversial issue. This paper is a contribution to the discussion on the interpretation of fetal skeleton variations in the context of risk assessment. Dose-response relationships of skeleton variations and malformations induced by three antineoplastic drugs (FUDR: 5-fluoro-2'-deoxyuridine, HU: hydroxyurea and 6-MPr: 6-mercaptopurine riboside) were evaluated. FUDR (0, 3, 14, 25, 35, 45, 55 and 65mg/kg body wt sc) and HU (0, 250, 300, 350, 400, 450, 500 and 550mg/kg body wt ip) were administered to rats on gestation day 11 (GD 11) while 6-MPr (0, 3, 7, 10 and 14mg/kg body wt sc) was given on GD 11, or on GD 12. Caesarean sections were performed on GD 21 and all fetuses were cleared and stained with alizarin red S for skeleton examination. Drugs given on GD 11 increased the incidence of thoracic and lumbar vertebra (dumbbell-shaped and bipartite ossification center (o.c.) and sternum (misaligned sternebrae) variations in a dose-dependent manner. Occurrence of zygomatic bone fused with maxilla (a variation in our rats) was also increased by HU and 6-MPr (GD 11) but it was not altered by FUDR. Spontaneous occurrence of wavy ribs was reduced by all treatments. Malformations such as cleft palate, tympanic bone absent and tibia absent were also increased in a dose-dependent manner by the three compounds. No observed effect levels (NOEL) for variations, irrespective of the compound administered, were generally lower than NOELs for malformations. In the discussion, we supported the view that any dose-related increase in the incidence of variations should be taken into account for determination of NOELs in routine studies. Increased occurrences of skeleton variations in term fetuses are also to be considered in risk assessment, unless experimental evidence exists that a particular change has no detrimental effect on the animal survival or health after birth or that it does not occur in humans. PMID- 19682678 TI - Polychlorinated biphenyl and organochlorine pesticide contamination signatures in deep-sea fish from the Mediterranean Sea. AB - Polychlorinated biphenyl (PCB) and OCP concentrations were determined in the livers of two deep-sea fish species, roughsnout grenadier and hollowsnout grenadier, from the Adriatic Sea. In both species, contaminant concentrations were in the following order: PCBs>DDTs>>HCB. Contaminant load was higher in roughsnout grenadier (PCB: 12,327ngg(-1); DDTs: 5357ngg(-1); HCB: 13.1ngg(-1)) than in hollowsnout grenadier (PCB: 1234ngg(-1); DDTs: 763ngg(-1); HCB 6.3ngg( 1)). PCB patterns were dominated by higher chlorinated congeners (hexa-CBs: 50.3 52.1%, hepta-CBs: 29.6-35.5%, penta-CBs: 8.0-11.1% and octa-CBs :5.2-5.4%). PCBs 138, 153 180 and 187 were the most abundant. Regarding the DDT pattern, p,p'-DDE was prevalent in both species (roughsnout grenadier: 99.7%, hollowsnout grenadier: 90%), suggesting no recent DDT input. In both species, the total 2,3,7,8-tetrachlorodibenzo-p-dioxin toxic equivalent (TEQ) concentrations (roughsnout grenadier: mean 43.77pg/g, hollowsnout grenadier: mean 20.49pg/g), calculated from non- and mono-ortho PCBs, reached those encountered in marine organisms at higher levels in the trophic chain. PMID- 19682675 TI - White matter microstructural differences linked to left perisylvian language network in children with dyslexia. AB - Studies of dyslexia using diffusion tensor imaging (DTI) have reported fractional anisotropy (FA) differences in left inferior frontal gyrus (LIFG) and left temporo-parietal white matter, suggesting that impaired reading is associated with atypical white matter microstructure in these regions. These anomalies might reflect abnormalities in the left perisylvian language network, long implicated in dyslexia. While DTI investigations frequently report analyses on multiple tensor-derived measures (e.g., FA, orientation, tractography), it is uncommon to integrate analyses to examine the relationships between atypical findings. For the present study, semi-automated techniques were applied to DTI data in an integrated fashion to examine white matter microstructure in 14 children with dyslexia and 17 typically developing readers (ages 7-16 years). Correlations of DTI metrics (FA and fiber orientation) to reading skill (accuracy and speed) and to probabilistic tractography maps of the left perisylvian language tracts were examined. Consistent with previous reports, our findings suggest FA decreases in dyslexia in LIFG and left temporo-parietal white matter. The LIFG FA finding overlaps an area showing differences in fiber orientation in an anterior left perisylvian language pathway. Additionally, a positive correlation of FA to reading speed was found in a posterior circuit previously associated with activation on functional imaging during reading tasks. Overall, integrating results from several complementary semi-automated analyses reveals evidence linking atypical white matter microstructure in dyslexia to atypical fiber orientation in circuits implicated in reading including the left perisylvian language network. PMID- 19682679 TI - Combined gain and phase margins. AB - In this note, the limitations of conventional or separate gain or phase margin are shown. Then, the combined gain and phase margins are introduced to overcome the limitations and suit both stable and unstable systems. A simple method for their computation is presented and their applications in stabilization is demonstrated. PMID- 19682680 TI - Current management of isolated sternal fractures in the UK: time for evidence based practice? A cross-sectional survey and review of literature. AB - Routine admission of patients with isolated sternal fractures for observation is still widespread in the UK. However, the evidence appears to suggest that this is unnecessary. We undertook a cross-sectional telephone survey of management of isolated sternal fractures in the UK. We contacted 85 acute admitting units over a three-month period and were able to get a response from 67 units. Most of the hospitals were district general hospitals (52) and situated in England (49). The orthopaedic department was the most common admitting department. 51 units indicated that they regularly admit isolated sternal fractures for observation. Other indications for admission included pain control (33), abnormal cardiac enzymes (28), social circumstances (23), abnormal electrocardiogram (6), and low oxygen saturation (5). Chest X-ray was performed on admission in all hospitals. 57 hospitals performed ECG and cardiac enzyme tests prior to admission and 6 hospitals carried out echocardiogram following admission on a regular basis. Patients were not followed up on discharge. 2 hospitals with on-site cardiothoracic unit followed-up patients on discharge, and 1 hospital advised GP follow-up. A review of the literature indicated that patients with isolated sternal fractures are at low risk of significant cardiac, pulmonary or mediastinal complications and do not need extensive investigations or routine admission. The current practice of management of isolated sternal fractures in the UK does not appear to conform to available evidence. In order to decide on management plans based on more rigorous evidence, there is a need for a prospective double blind randomised study of patients with isolated sternal fractures, comparing those discharged to those admitted over a longer follow-up period. PMID- 19682681 TI - The effect of exercise on back pain and lordosis in pregnant women. PMID- 19682682 TI - Vulvar lymphangioma circumscriptum. PMID- 19682683 TI - Comparison of transabdominal versus transvaginal ultrasound to measure thickness of the lower uterine segment at term. AB - OBJECTIVE: To compare the accuracy of transvaginal (TVS) versus transabdominal (TAS) ultrasound to assess the thickness of the lower uterine segment (LUS). METHODS: Eighty-three pregnant women admitted for an elective cesarean delivery were enrolled. LUS thickness was measured using both TVS and TAS prior to the cesarean. The actual thickness of the LUS was measured using a sterile metal ruler after the neonate had been delivered. RESULTS: Seventeen women had unscarred uteri (20.1%); 41 had had one previous cesarean (49.4%); and 25 had had two previous cesareans (30.1%). Mean thickness of the LUS measured after delivery was 7.58+/-1.3 mm in unscarred uteri; 5.09+/-1.4 mm for one cesarean; and 3.92+/ 1.1 mm for two cesareans (P<0.01). Actual thickness of the LUS showed a significant correlation with TVS among the total (r(s)=0.89); with unscarred uteri (r(s)=0.68); with 1 cesarean (r(s)=0.89); and 2 cesareans (r(s)=0.68), while with TAS the correlations were significant only with the total (r(s)=0.53) and 2 previous cesareans (r(s)=0.63) (P<0.01). CONCLUSION: TVS is a more accurate method of assessing the thickness of the LUS compared with TAS. PMID- 19682684 TI - [45th Congress of the American Society of Clinical Oncology (ASCO) Orlando, May 30th-June 2nd 2009]. PMID- 19682685 TI - [The strategy "wait and watch" in patients with a cancer of bottom stocking rectum with a complete clinical answer after neoadjuvant radiochemotherapy]. PMID- 19682686 TI - [A "cystic" lesion of the pancreas]. PMID- 19682687 TI - Plasma triglyceride levels and body mass index values are the most important determinants of prebeta-1 HDL concentrations in patients with various types of primary dyslipidemia. AB - OBJECTIVE: Experimental studies have shown that the prebeta-1 subclass of high density lipoprotein particles (prebeta-1 HDL) may play an important role in the reverse cholesterol transport pathway as the initial acceptors of cellular cholesterol. The aim of the present study was the direct comparison of prebeta-1 HDL values in individuals with various types of primary dyslipidemias. METHODS: Four hundred and eighty-six unrelated individuals were included in the study. According to their lipid values study participants were subdivided into four groups: control group (n=206), type IIA dyslipidemia group (n=148), type IIB dyslipidemia group (n=49) and type IV dyslipidemia group (n=83). RESULTS: All dyslipidemic patients displayed higher concentrations of prebeta-1 HDL compared to control individuals. However, patients with dyslipidemias characterized by an abnormal catabolism of triglyceride-rich lipoproteins (such as dyslipidemias of type IIB and IV) tend to have higher prebeta-1 HDL values compared to patients with hypercholesterolemia, and this increase is proportional to the degree of hypertriglyceridemia. In addition, patients with metabolic syndrome exhibited significantly higher levels of prebeta-1 HDL compared to individuals that do not fulfill the criteria for the diagnosis of this syndrome. Multiple regression analysis revealed that serum triglyceride concentrations and body mass index (BMI) values were the most important determinants of prebeta-1 HDL levels in our population. CONCLUSION: All dyslipidemic patients exhibit increased prebeta-1 HDL concentrations as compared to normolipidemic individuals. Whether this increase represents a defensive mechanism against atherosclerosis or it is indicative of impaired maturation of HDL particles and thus of a defective reverse cholesterol transport mechanism remains to be established. PMID- 19682688 TI - Tacrolimus-eluting stent inhibits neointimal hyperplasia via calcineurin/NFAT signaling in porcine coronary artery model. AB - AIMS: The purpose is to elucidate the mechanism by which a newly developed tacrolimus-eluting stent (TES) prevents neointimal hyperplasia after stenting. METHODS AND RESULTS: The three major coronary arteries in juvenile swine were randomized to implantation of either a TES or bare metal stent (BMS). Twelve weeks after stenting, the TES showed 29% less neointimal area than the BMS. Immunohistochemical staining showed that the expression of calcineurin was up regulated in the neointima and media after stenting, and the TES inhibited this up-regulation. Western blotting demonstrated that the expression of calcineurin, nuclear factor of activated T cell (NFAT), and interleukin-2 (IL-2) was lower with the TES than with the BMS. To confirm the effect of tacrolimus on vascular smooth muscle cells (VSMCs) and its mechanism, cultured rat VSMCs were incubated with 12.5 microM of tacrolimus (tacrolimus group) or without tacrolimus (control group). The cell number of the tacrolimus group was significantly lower than that of the control group at 48 h of incubation. Western blotting demonstrated that tacrolimus decreased the expression of calcineurin, NFATc4, and IL-2 of cultured VSMCs. We confirmed that calcineurin small-interfering RNA (siRNA) decreased cell proliferation and the expression of NFATc4 and IL-2 in cultured VSMCs compared with negative control-siRNA. CONCLUSION: The newly developed TES inhibited neointimal hyperplasia after stenting via the calcineurin/NFAT/IL-2 signaling pathway, which is one of several mechanisms through which TES inhibits restenosis. Calcineurin may be an important molecular target to prevent restenosis after stenting. PMID- 19682689 TI - Lipopolysaccharide up-regulates the expression of Fcalpha/mu receptor and promotes the binding of oxidized low-density lipoprotein and its IgM antibody complex to activated human macrophages. AB - Natural IgM antibodies against oxidized low-density lipoprotein (oxLDL) can inhibit the binding of oxLDL to macrophages and bacterial infection may deteriorate the pathogenesis of atherosclerosis. However, little is known about the molecular mechanisms underlying the action of bacterial lipopolysaccharide (LPS) in the binding of oxLDL to macrophages, contributing to the formation of foam macrophages. In this study, human monocytes-derived macrophages were cultured and incubated with purified human anti-oxLDL IgM antibodies (HAO-IgM), lipopolysaccharide (LPS) and oxLDL. The HAO-IgM were found specifically inhibited the binding of CuoxLDL to naive macrophages but failed to inhibit the binding of CuoxLDL to LPS-activated macrophages and promoted the formation of CuoxLDL mediated foam macrophages. Furthermore, the HAO-IgM F(ab')(2) or pre-incubation with unrelated IgM inhibited the binding of HAO-IgM/CuoxLDL complex to LPS activated macrophages, suggesting that Fcalpha/mu receptor (Fcamr) may be responsible for the binding of HAO-IgM/CuoxLDL complex to LPS-activated macrophages. Indeed, LPS up-regulated the expression of Fcamr in macrophages in a dose- and time-dependent manner, which was diminished by treatment with anti TLR4. In addition, LPS induced the phosphorylation of p38MAPK and translocation of NF-kappaB p65, contributing to the up-regulated expression of Fcamr in macrophages as treatment with specific inhibitor for p38MAPK (SB203580) or NF kappaB (PDTC) attenuated the up-regulation of Fcalpha/mu receptor expression induced by LPS in macrophages. Inhibition of p38MAPK and NF-kappaB decreased the foam cells formation increased by Fcamr expression. These data demonstrated that LPS, through the TLR4 receptor, activated the p38MAPK and NF-kappaB pathways and up-regulate the expression of Fcamr in human macrophages, which promotes the binding of IgM/CuoxLDL complex to macrophages and the formation of foam cells. Therefore, our findings provide a new explanation why bacterial infection deteriorates the pathogenesis of atherosclerosis. PMID- 19682691 TI - Wearing shoes increasing dorsiflexion improves short-term but not long-term balance control in young healthy adults. AB - Wearing sport shoes inducing ankle dorsiflexion has been shown to alter the biomechanical specificities of the stretched muscles. The possible effect over the short and long term upon the sensorial capacities induced by such stretching has not been addressed yet. Fourteen healthy individuals were involved to assess the proprioceptive repercussion and their effects upon postural control strategies. Postural control and proprioceptive assessment were measured twice: when receiving sport shoes inducing ankle dorsiflexion and 18 days later. Proprioceptive effects were assessed using an ad-hoc device through which the seated and blindfolded subjects were required to reposition their feet in a starting position after the ankles were passively displaced to dorsiflexed and plantarflexed positions. Center-of-gravity horizontal displacements (CG(v)), estimated from center-of-pressure (CP) displacements, and CP-CG(v) displacements were measured through a force platform during upright quiet stance maintenance. The initial session was recorded with the subjects barefoot and wearing the shoes with a set of chocks with 0 degrees (horizontal) and -5 degrees (dorsiflexion) tilting angles. The second session included only barefoot performance in horizontal and dorsiflexion conditions. Dorsiflexion had no immediate effect on the postural control strategies along the anteroposterior axis. In contrast, barefoot or wearing shoes, stability was increased along the mediolateral axis during the dorsiflexion conditions. No ankle proprioceptive or postural change was observed after wearing the shoes for 18 days. Wearing dorsiflexion sport shoes induces short-term effects probably by inducing a backward tilt of the pelvis. A muscular adaptation likely prevents this effect from being prolonged. PMID- 19682690 TI - Fatigue creep damage at the cement-bone interface: an experimental and a micro mechanical finite element study. AB - The goal of this study was to quantify the micromechanics of the cement-bone interface under tensile fatigue loading using finite element analysis (FEA) and to understand the underlying mechanisms that play a role in the fatigue behavior of this interface. Laboratory cement-bone specimens were subjected to a tensile fatigue load, while local displacements and crack growth on the specimen's surface were monitored. FEA models were created from these specimens based upon micro-computed tomography data. To accurately model interfacial gaps at the interface between the bone and cement, a custom-written erosion algorithm was applied to the bone model. A fatigue load was simulated in the FEA models while monitoring the local displacements and crack propagation. The results showed the FEA models were able to capture the general experimental creep damage behavior and creep stages of the interface. Consistent with the experiments, the majority of the deformation took place at the contact interface. Additionally, the FEA models predicted fatigue crack patterns similar to experimental findings. Experimental surface cracks correlated moderately with FEA surface cracks (r(2)=0.43), but did not correlate with the simulated crack volume fraction (r(2)=0.06). Although there was no relationship between experimental surface cracks and experimental creep damage displacement (r(2)=0.07), there was a strong relationship between the FEA crack volume fraction and the FEA creep damage displacement (r(2)=0.76). This study shows the additional value of FEA of the cement-bone interface relative to experimental studies and can therefore be used to optimize its mechanical properties. PMID- 19682692 TI - How do spinal segments move? AB - PURPOSE: To study and clarify the kinematics of spinal segments following cyclic torques causing axial rotation (T(z) (t)), lateral-flexion (T(x) (t)), flexion/extension (T(y) (t)). METHODS: A 6D--Measurement of location, alignment, and migration of the instantaneous helical axis (IHA) as a function of rotational angle in cervical, thoracic, and lumbar segments subjected to axially directed preloads. RESULTS: IHA retained an almost constant alignment, but migrated along distinct centrodes. THORACIC SEGMENTS: IHA was almost parallel to T(z) (t), T(x) (t), or T(y) (t), stationary for T(x) (t) or T(y) (t), and migrating for T(z) (t) along dorsally opened bows. IHA locations hardly depended on the position or size of axial preload. LUMBAR SEGMENTS: IHA was also almost parallel to T(z) (t), T(x) (t), or T(y) (t). In axial rotation IHA-migration along wide, ventrally or dorsally bent bows depending on segmental flexional/extensional status. Distances covered: 20-60mm. In lateral-flexion: IHA-migration to the left/right joint and vice versa. In flexion/extension IHA-migration from the facets to the centre of the disc. CERVICAL SEGMENTS: In flexion/flexion IHA was almost stationary for and parallel to T(y) (t). In axial rotation or lateral-flexion IHA intersected T(z) (t)/T(x) (t) under approximately -30 degrees /+30 degrees. CONCLUSIONS: Generally joints alternate in guidance. Lumbar segments: in axial rotation and lateral flexion parametrical control of IHA-position and IHA-migration by axial preload position. Cervical segments: kinematical coupling between axial rotation and lateral-flexion. The IHA-migration guided by the joints should be taken into account in the design of non-fusion implants. FE-calculations of spinal mechanics and kinematics should be based on detailed data of curvature morphology of the articulating surfaces of the joint facets. PMID- 19682693 TI - Footfall dynamics for racewalkers and runners barefoot on compliant surfaces. AB - The main purpose of this study is to investigate the role of footfall surface compliance on the physical parameters affecting barefoot racewalkers and runners. These parameters are identified using a new inverted pendulum body model with a forward moving foot pivot. Model correlations of footfall loads measured for four compliant surface mats showed leg-foot compression stiffness for both gaits were in the range of 10.8-12.9 kN/m, with the initial stiffness spikes in the range of 6.5-52 kN/m. The average leg damping factor was about 0.6% for racewalkers and 6% for runners. For both gaits there was negative leg damping just prior to foot lift-off. Compared to the peak reactions for the rigid surface, a mat of intermediate compliance (1020 kN/m) was effective in reducing the runners' peak reaction spikes by as much as 17%. PMID- 19682694 TI - Gait posture estimation using wearable acceleration and gyro sensors. AB - A method for gait analysis using wearable acceleration sensors and gyro sensors is proposed in this work. The volunteers wore sensor units that included a tri axis acceleration sensor and three single axis gyro sensors. The angular velocity data measured by the gyro sensors were used to estimate the translational acceleration in the gait analysis. The translational acceleration was then subtracted from the acceleration sensor measurements to obtain the gravitational acceleration, giving the orientation of the lower limb segments. Segment orientation along with body measurements were used to obtain the positions of hip, knee, and ankle joints to create stick figure models of the volunteers. This method can measure the three-dimensional positions of joint centers of the hip, knee, and ankle during movement. Experiments were carried out on the normal gait of three healthy volunteers. As a result, the flexion-extension (F-E) and the adduction-abduction (A-A) joint angles of the hips and the flexion-extension (F E) joint angles of the knees were calculated and compared with a camera motion capture system. The correlation coefficients were above 0.88 for the hip F-E, higher than 0.72 for the hip A-A, better than 0.92 for the knee F-E. A moving stick figure model of each volunteer was created to visually confirm the walking posture. Further, the knee and ankle joint trajectories in the horizontal plane showed that the left and right legs were bilaterally symmetric. PMID- 19682695 TI - High-temperature liquid chromatography. Part I. Determination of the vapour pressures of binary solvent mixtures--implications for liquid chromatographic separations. AB - This paper is the first in a series of consecutive publications, explaining the concept of high-temperature liquid chromatography under various important aspects. The first publication deals with the determination of the vapour pressure of binary solvent mixtures used in reversed-phase liquid chromatography. Also, the practical implications regarding the use of elevated temperatures in liquid chromatography and towards commercially available HPLC systems are explained. PMID- 19682696 TI - Sulfonated polyvinyl chloride fibers for cation-exchange microextraction. AB - Polyvinyl chloride (PVC) fiber was derivatized by concentrated sulfuric acid to yield sulfonated PVC (PVC-SO3H). The PVC-SO3H fiber had dual properties as a sorbent, based on cation-exchange and hydrophobicity. In the present study, the novel fiber was used directly as an individual device for extraction purposes in the cation-exchange microextraction of anaesthetics, followed by high-performance liquid chromatography-UV analysis. The results demonstrated that this PVC-SO3H fiber-based microextraction afforded convenient operation and cost-effective application to basic analytes. The limits of detection for four anaesthetics ranged from 1.2 to 6.0 ng/mL. No carryover (because of its disposable usage), and no loss of sorbent phase (which normally occurs in stir-bar sorptive extraction) during extraction were observed. PMID- 19682697 TI - Evaluation of multiplexed hollow fiber flow field-flow fractionation for semi preparative purposes. AB - A multiplexed hollow fiber flow field-flow fractionation (MxHF5) is introduced to increase throughput of an HF5 channel system for semi-preparative purposes. HF5, a cylindrical version of the flow field-flow fractionation (FlFFF) operated with a porous, hollow fiber membrane by controlling the ratio of radial and axial flow rates, is capable of fractionating proteins, cells, and macromolecules by size. An advantage of HF5 is its inexpensive channel construction, allowing for disposability that can reduce run-to-run carryover problems. MxHF5 constructed in this study was made with six parallel HF5 modules connected to seven-port manifolds for the semi-preparative scale separation of proteins or biological particles. For the evaluation of MxHF5 separation efficiency, protein standards were utilized to test peak recoveries, band broadening, and throughput. The assembly showed the possibility of handling up to 50 microg of proteins without incurring overloading. The developed channel was applied to demonstrate size sorting of lipoproteins for the future study of size dependent lipidomic and proteomic analysis. The current trial offers a unique advantage of scaling up HF5 separation without using wide-bore, hollow fibers which sacrifice separation speed. PMID- 19682698 TI - Bilinear electric field gradient focusing. AB - Electric field gradient focusing (EFGF) uses an electric field gradient and a hydrodynamic counter flow to simultaneously separate and focus charged analytes in a channel. Previously, most EFGF devices were designed to form a linear field gradient in the channel. However, the peak capacity obtained using a linear gradient is not much better than what can be obtained using conventional CE. Dynamic improvement of peak capacity in EFGF can be achieved by using a nonlinear gradient. Numerical simulation results indicate that the peak capacity in a 4-cm long channel can be increased from 20 to 150 when changing from a linear to convex bilinear gradient. To demonstrate the increased capacity experimentally, an EFGF device with convex bilinear gradient was fabricated from poly(ethylene glycol) (PEG)-functionalized acrylic copolymers. The desired gradient profile was confirmed by measuring the focusing positions of a standard protein for different counter flow rates at constant voltage. Dynamically controlled elution of analytes was demonstrated using a monolith-filled bilinear EFGF channel. By increasing the flow rate, stacked proteins that were ordered but not resolved after focusing in the steep gradient segment were moved into the shallow gradient segment, where the analyte peak resolution increased significantly. In this way, the nonlinear field gradient was used to realize a dynamic increase in the peak capacity of the EFGF method. PMID- 19682699 TI - Characterization of ethylene oxide-propylene oxide block copolymers by combination of different chromatographic techniques and matrix-assisted laser desorption ionization time-of-flight mass spectroscopy. AB - Block copolymers of ethylene oxide (EO) and propylene oxide (PO) are characterized by combination of two-dimensional chromatography and MALDI-TOF-MS. Liquid chromatography under critical conditions (LCCC) is used as first dimension and fractions are collected, mobile phase evaporated and diluted in the mobile phase used in second dimension (SEC or LAC). This two-dimensional chromatography in combination of MALDI-TOF-MS gives information about purity of reaction products, presence of the byproducts, chemical composition and molar mass distribution of all the products. PMID- 19682700 TI - Multifunctional nanoparticles/silica microsphere assemblies using polyglycidyl methacrylate shells as supports. AB - A general method to prepare functional (or multifunctional) nanoparticles/silica microsphere assemblies is reported in this article. A thin shell of polyglycidyl methacrylate is grafted on the surface of silica through surface-initiated atom transfer radical polymerization technique. And then, various types of nanoparticles, including water-soluble CdTe quantum dots, Au nanoparticles and oil-soluble Fe(3)O(4) nanoparticles are assembled on silica microspheres, respectively, or simultaneously. The properties of the assembled nanoparticles are well retained in the nanocomposite assemblies, and the controllable integration of magnetic and fluorescent properties can be achieved through varying the proportion of different nanoparticles assembled on nanoparticle/silica microsphere. PMID- 19682702 TI - Comparison of surface properties between kaolin and metakaolin in concentrated lime solutions. AB - The surface adsorption of calcium hydroxide onto kaolin and metakaolin was investigated by monitoring with atomic emission spectroscopy and pH measurements the amounts of ions left in solution after exposing clays to calcium hydroxide solutions of various concentrations. Both clays adsorb calcium and hydroxyl ions but differently. Kaolin adsorbs calcium hydroxide not only at the edges of the clay particles but also onto the basal faces. The adsorbed hydrated calcium ions form a layer on the clay particle surfaces, preventing further dissolution of the clay mineral platelet. Metakaolin shows high pozzolanic activity, which provides the quick formation of hydrated phases at the interfaces between metakaolin and lime solutions. The nature of the hydration products has been investigated using X-ray diffraction (XRD) and differential thermal analysis (DTA). The most important hydrated phases like CSH (hydrated calcium silicate) and C(2)ASH(8) (gehlenite) have been identified. PMID- 19682701 TI - Phase behavior of bile acid/lipid/water systems containing model dietary lipids. AB - Dietary lipids are solubilized in bile acid micelles in the small intestine. In the present study, we investigate the phase behavior of bile acid/model rapeseed oil (or model beef tallow)/water systems to predict interfacial phenomena during consumption of a variety of foods. The structures of molecular assemblies are identified based on polarizing microscope images, wide-angle X-ray diffraction (WAXD) and small-angle X-ray scattering (SAXS). The results of in vitro tests suggest that an increase in the intake of model rapeseed oil causes the formation of multi-lamellar vesicles and lamellar liquid crystals. The molecules in the lamellar liquid crystal are formed highly ordered layer structure with the spacing of 8.8nm along the c-axis, while monoclinic packed structure is constructed as two-dimensional structure in ab-plane due to bulky molecular structures of bile acid and unsaturated fatty acid. When the model beef tallow composition in the model system is more than several wt.%, stearic acid crystals are extracted. Moreover, bicarbonate ions are important ingredients to solubilize >10wt.% of the model lipids. These phase transitions might be induced by the addition of dietary lipids in vivo during the consumption of oil or meat. Our findings are significant for understanding the lipid absorption process in the small intestine, and for developing medical and healthcare products. PMID- 19682704 TI - Gels from a semifluorinated n-alkane in fluorinated solvents as a probe for intermolecular interactions. AB - Diblock semifluorinated n-alkanes can form aggregates and gels in fluorinated solvents. We have investigated the thermal behavior of binary mixtures comprising F(CF(2))(8)(CH(2))(16)H and fluorinated solvents. The solvents were perfluorohexane, perfluoroheptane, perfluorooctane, perfluorooctyl bromide, perfluorodecalin, and perfluorotributylamine. The phase diagrams were used to calculate the activity coefficients of the two components and the main excess thermodynamic functions. The solubility and self-assembly behavior of F(8)H(16) in the fluorinated solvents are related to the different solute-solvent dispersion interactions that depend on the polarizabilities and ionization potentials of the interacting species, and on the structural properties of the solvent. PMID- 19682703 TI - Nano-structured calcium silicate hydrate functionalised with iodine. AB - Nano-structured calcium silicate hydrate can physisorb or chemisorb iodine, making it interesting for medical or materials science applications, where a slow, controlled release of iodine is desired. It was found that iodine can be sorbed and released by applying the elemental halogen in solution, either as a gas or as a solid. At ambient temperatures the sorption and desorption process is quantitative and physical, meaning that the same amount of iodine is taken up and released. At temperatures above 32.5 degrees C (305.7K) iodine reacts with the calcium silicate hydrate forming a complex, which is stable above the sublimation temperature of iodine. The formation energy for the iodine calcium silicate hydrate complex was established to be 41.8+/-0.8kJmol(-1) by calorimetry and the nature of the complex was investigated using X-ray photoelectron spectroscopy. PMID- 19682705 TI - Degradation of phenol by mechanical activation of a rutile catalyst. AB - In the present paper a novel mechanochemical process for the elimination of organic pollutants dissolved in water is proposed. In this regard, phenol aqueous solutions (100mgL(-1)) were ball-milled for 0, 12, 18, 24, 36, 48, and 72h with and without a well-characterized (XRD, SEM, and N(2) Adsorption), rutile powder catalyst and the reaction products analyzed with UV and GC/MS. It was found that when the catalyst was not included in the process, phenol was not affected, but when it was included, phenol was decomposed. The catalyst itself did not change and the reaction follows a pseudo-first-order kinetics. Besides, intermediates which are characteristic of the ()OH radical mechanism were found in the reaction products. Then, a mechanism similar to those accepted for other advanced oxidation processes was proposed. The value measured for the pseudo-first-order reaction constant was very low, indicating that the reported process is inefficient. Nevertheless, this problem could be solved by applying catalysts consisting of particles with smaller diameters. PMID- 19682706 TI - Type II bidirectional ventricular tachycardia in a patient with myocardial infarction. AB - A 84-year-old man presented to the emergency department complaining of chest pain and palpitations. He had no history of coronary artery disease. The 12-lead electrocardiography showed bidirectional ventricular tachycardia (BVT). Coronary angiography revealed severe mid left anterior descending and mid left circumflex lesions. The BVT, in this case, was most likely due to myocardial ischema. The ethiology of published BVT cases are most commonly digitalis toxicity and rarely herbal aconitine poisoning, hypokalemic periodic paralysis, cathecolaminergic VT, myocarditis, and Anderson-Tawil syndrome. The patient had neither of these underlying conditions. To the best of our knowledge and research in the literature, there was no report of bidirectional VT in the patients with myocardial infarction. PMID- 19682707 TI - Potential danger of ocular compression in paroxysmal supraventricular tachycardia in patients with latent preexcitation. AB - The vagal maneuver is the first line of therapeutic available for patients with paroxysmal supraventricular tachycardia. It increases vagal tone and includes the traditional ocular compression, carotid sinus massage, and Valsalva maneuver. A 40-year-old man was admitted because of 180 beats/min regular narrow QRS-complex tachycardia. The physician in the emergency department had performed an ocular compression, and at its ending, the tachycardia degenerated into unstable hemodynamically high ventricular rate atrial fibrillation. It was reverted to sinus rhythm by electrical shock. The electrophysiologic study documented a latent posterolateral bypass tract, with an anterograde refractory period of 210 milliseconds, which was successfully ablated. PMID- 19682708 TI - Correlations between the atherosclerotic changes of femoral, carotid and coronary arteries: a post mortem study. AB - OBJECTIVE: To compare the severity of atherosclerosis in the carotid, coronary and femoral arteries in autopsy findings of stroke patients. METHODS AND RESULTS: 40 patients (age: 75.2 (12.3) years, 21 men, 19 women) were investigated, who died of ischemic stroke. Carotid, femoral and coronary arteries were removed and cut into slices. Atherosclerotic changes were scored and compared. The severity of atherosclerotic changes of the common carotid artery did not correlate with any other arteries. Atherosclerotic parameters of the internal carotid artery correlated with those of the deep femoral and common femoral arteries (r=0.457 0.459; P=0.022-0.028 respectively). We found significant correlations between the deep femoral artery and left anterior descendent coronary arteries (r=0.513; P=0.012). External carotid artery correlated with both the left anterior descendent coronary and deep femoral arteries (r=0.458-0.473 and P=0.028-0.017 respectively). CONCLUSIONS: The severity of atherosclerosis in the external carotid arteries and/or the femoral arteries showed a stronger correlation with the atherosclerosis in the coronaries than that of the common carotid arteries. PMID- 19682709 TI - Ischemic stroke patterns and hemodynamic features in patients with small vertebrobasilar artery. AB - BACKGROUND: To determine the role of small vertebrobasilar artery (SVBA) in patients with posterior circulation stroke (PCS), we evaluated the ischemic patterns, collateral features, and stroke mechanisms in PCS patients with SVBA. METHODS: Ischemic findings on magnetic resonance (MR) imaging were correlated with 3D time-of-flight/contrast-enhanced MR angiography and/or catheter angiography in 18 patients (mean age, 68.0+/-11.8 years; 9 males). SVBA (lumen diameter of <3 mm) was compared with stenotic normal-sized VBA (NVBA) in 14 PCS patients. RESULTS: Ischemic lesions were predominantly observed in the cerebellum and/or medulla (vertebral artery (VA) territory). All subjects had fetal posterior circulation (FPC) from the internal carotid artery to the posterior cerebral artery. Sixteen patients (88.9%) had distal or diffuse VA stenosis/occlusion. In 14 patients (77.8%), long circumferential artery (LCA) was prominently observed. In atherothrombotic patients, infratentorial PCS might occur following artery-to-artery embolism from the low-flowed or stenotic VA to LCA. Ischemic patterns between subjects with and without VA disease were almost similar. As the degree of VA disease increased, the frequency of LCA prominence showed an increased tendency (P=0.003). Relatively small, scattered infarcts were observed in patients with SVBA than in those with stenotic NVBA. CONCLUSIONS: FPC does not protect against infratentorial PCS. Regardless of extensive arterial lesions, relatively small infarcts may be due to previously established collaterals from the LCA, which could compensate for the defects in the infratentorial area. PMID- 19682710 TI - Reactivity of transparent exopolymeric particles: a key parameter of trace metal cycling in the lagoon of Noumea, New Caledonia. AB - Seawater samples were collected in the lagoon of Noumea (New Caledonia) along two transects from coastal bays to the oligotrophic barrier reef. Dissolved (<0.2 microm) and sub-micrometer (0.2-0.8 microm) concentrations of chromium, nickel and zinc were measured. Concentrations in the 0.2-0.8 microm size range were correlated with transparent exopolymeric particles (TEP) turnover rates, suggesting that a TEP pool rapidly cycling in the system is more reactive toward metals. The role of TEP reactivity in the transfer efficiency of metals from dissolved to particulate phases was estimated through variations of the metal partitioning coefficient K between sub-micrometer TEP and solution, as a function of the water mass residence time. The log K (6.0-8.0) increased from high to low residence time, suggesting that TEP reactivity increases when water masses renew rapidly. This suggests that hydrodynamics control TEP reactivity toward metals. The characteristics (adsorption vs. complexation) of TEP-metal association are discussed. PMID- 19682711 TI - Monitoring the freely dissolved concentrations of polycyclic aromatic hydrocarbons (PAH) and alkylphenols (AP) around a Norwegian oil platform by holistic passive sampling. AB - In order to assess the environmental impact of aquatic discharges from the offshore oil industry, polar organic chemical integrative samplers (POCIS) and semipermeable membrane devices (SPMDs) were deployed around an oil platform and at reference locations in the North Sea. Exposure to polycyclic aromatic hydrocarbons (PAH) and alkylated phenols (AP) was determined from passive sampler accumulations using an empirical uptake model, the dissipation of performance reference compounds and adjusted laboratory derived sampling rates. Exposure was relatively similar within 1-2 km of the discharge point, with levels dominated by short chained C1-C3 AP isomers (19-51 ngL(-1)) and alkylated naphthalenes, phenanthrenes and dibenzothiophenes (NPD, 29-45 ngL(-1)). Exposure stations showed significant differences to reference sites for NPD, but not always for more hydrophobic PAH. These concentrations are several orders of magnitude lower than those reported to give both acute and sub-lethal effects, although their long term consequences are unknown. PMID- 19682712 TI - Metal accumulation in sea urchins and their kelp diet in an Arctic fjord (Kongsfjorden, Svalbard). PMID- 19682713 TI - Identification and quantification of mixed sources of oil spills based on distributions and isotope profiles of long-chain n-alkanes. AB - Combined with quantitative determination of concentration and isotopic composition of petroleum hydrocarbons, weathering simulation experiments on artificially mixed oils and their two end-member oils are performed for identification and quantification of mixed sources. The >C(18)n-alkanes show no appreciable losses during a short-term weathering process. An approach based on distribution of long-chain n-alkanes (>C(18)) is suggested for estimating the contribution proportion of each source in mixed oils. Stable carbon isotope profile of individual n-alkanes is a powerful tool to differentiate sources of oil spills, but unavailable to accurately allocate each contribution due to a relatively large analytical error. PMID- 19682715 TI - Lecithin cholesterol acyltransferase (LCAT) activity as a predictor for ketosis and parturient haemoglobinuria in Egyptian water buffaloes. AB - Lecithin cholesterol acyltransferase (LCAT) activity was measured in 48 Egyptian water buffaloes four weeks pre-parturient. The activity was significantly low in 37 buffaloes (77.1%). Four weeks post-partum, clinical examination revealed that 23 buffaloes had the clinical signs of ketosis (K) while 14 had the clinical signs of parturient-haemoglobinuria (PHU). Serum samples were collected from 5 buffaloes of each group (K and PHU) besides 5 clinically healthy buffaloes with normal LCAT (control). Glucose level was significantly reduced in K and PHU groups while the phosphorous (P) level was significantly reduced in PHU group compared to control. There were significant reductions in the total cholesterol, free cholesterol, triglycerides, total protein and albumin in K and PHU groups; whereas, significant increases in AST, GGT, non-esterified fatty acids (NEFA) and beta-hydroxybutyric acid (BHBA) in K and PHU groups were detected. Therefore, LCAT could be a predictor for metabolic disorders in Egyptian water buffaloes. PMID- 19682714 TI - Elderberry flavonoids bind to and prevent H1N1 infection in vitro. AB - A ionization technique in mass spectrometry called Direct Analysis in Real Time Mass Spectrometry (DART TOF-MS) coupled with a Direct Binding Assay was used to identify and characterize anti-viral components of an elderberry fruit (Sambucus nigra L.) extract without either derivatization or separation by standard chromatographic techniques. The elderberry extract inhibited Human Influenza A (H1N1) infection in vitro with an IC(50) value of 252+/-34 microg/mL. The Direct Binding Assay established that flavonoids from the elderberry extract bind to H1N1 virions and, when bound, block the ability of the viruses to infect host cells. Two compounds were identified, 5,7,3',4'-tetra-O-methylquercetin (1) and 5,7-dihydroxy-4-oxo-2-(3,4,5-trihydroxyphenyl)chroman-3-yl-3,4,5 trihydroxycyclohexanecarboxylate (2), as H1N1-bound chemical species. Compound 1 and dihydromyricetin (3), the corresponding 3-hydroxyflavonone of 2, were synthesized and shown to inhibit H1N1 infection in vitro by binding to H1N1 virions, blocking host cell entry and/or recognition. Compound 1 gave an IC(50) of 0.13 microg/mL (0.36 microM) for H1N1 infection inhibition, while dihydromyricetin (3) achieved an IC(50) of 2.8 microg/mL (8.7 microM). The H1N1 inhibition activities of the elderberry flavonoids compare favorably to the known anti-influenza activities of Oseltamivir (Tamiflu; 0.32 microM) and Amantadine (27 microM). PMID- 19682716 TI - [Giant cell tumors and pseudogiant cell tumors of the jaws]. AB - Giant cell tumors and pseudogiant cell tumors of the jaws are divided into cherubism, central giant cell granuloma (CGCG), aneurysmal cyst, giant cell tumor, and hyperparathyroidism-jaw tumor. These tumors have variable clinical characteristics, such as bilateral tumors in cherubism, disseminated tumors in hyperparathyroidism, or focal tumors in other types. Their radiological presentation is quite similar with presence of well-delimitated uni- or multilocular radiolucency. The primary surgical treatment must be conservative. Anatomopathology gives the histologic diagnosis, with presence of giant cells. Some tumors are locally aggressive leading to larger bone exeresis. The nosology of these giant-cell tumors remains unclear, especially for giant cell central granuloma and giant cell tumors. PMID- 19682717 TI - Middle pancreatectomy: safety and long-term results. AB - BACKGROUND: Pancreaticoduodenectomy and distal pancreatectomy for lesions of the neck or body of the pancreas sacrifice a large amount of normal pancreatic tissue. Middle pancreatectomy (MP) is a parenchyma sparing technique that reduces the risk of postoperative endocrine and exocrine insufficiency. This study aims to evaluate the perioperative and long-term results of MP and to clarify whether MP can be performed with outcomes comparable with traditional pancreatectomies. METHOD: Twenty-six patients who underwent MP for benign or low-grade malignant tumor of the pancreas between 1991 and 2006 at the Department of Surgery II, Nagoya University Graduate School of Medicine, were identified. Their outcomes were compared with 2 separate control groups, 35 left-side pancreatectomies (LSP) and 60 right-side pancreatectomies (RSP). RESULTS: The mean operating time of the MP group was 295 minutes, which was significantly shorter than that for RSP (P=.0001). The rate of pancreatic fistula formation was higher in the MP group than in the 2 control groups, although the differences did not reach statistical significance. After a mean follow-up of 71 months, postoperative endocrine function was equivalent to the pre-operative values in the MP group, and none of the patients developed diabetes mellitus postoperatively. Only 1 patient in the MP group required enzyme substitution postoperatively for exocrine insufficiency. The MP group was inclined to be superior to the other 2 control groups in terms of postoperative nutritional status. CONCLUSION: Middle pancreatectomy is a reasonable technique that is indicated for selected patients with benign or low malignant tumors in the neck and body of the pancreas. Middle pancreatectomy seems to result in better preservation of exocrine and endocrine functions as well as in better nutritional status postoperatively. PMID- 19682718 TI - Enhanced experimental approach to measure the absolute ultrasonic wave attenuation. AB - The absolute ultrasonic wave attenuation is an important input parameter for mathematical models, which play an increasingly important role in non-destructive testing. The measurement of the absolute ultrasonic wave attenuation however is a difficult task. When conventional measurement techniques are applied, corrections to the raw data are required to account for apparent losses. In this study, a modified experimental approach is proposed to determine the absolute ultrasonic attenuation without any further corrections of the raw data. PMID- 19682719 TI - Ultrasonic particle size fractionation in a moving air stream. AB - Identification of bio-aerosol particles may be enhanced by size sorting before applying analytical techniques. In this paper, the use of ultrasonic acoustic radiation pressure to continuously size fractionate particles in a moving air stream is described. Separate particle-laden and clean air streams are introduced into a channel and merged under laminar flow conditions. An ultrasonic transducer, mounted flush to one wall of the channel, excites a standing ultrasonic wave perpendicular to the flow of the combined air stream. Acoustic radiation forces on the particles cause them to move transverse to the flow direction. Since the radiation force is dependent upon the particle size, larger particles move a greater transverse distance as they pass through the standing wave. The outlet flow is then separated into streams, each containing a range of particle sizes. Experiments were performed with air streams containing glass microspheres with a size distribution from 2-22 microm, using a centerline air stream velocity of approximately 20 cm/s. An electrostatic transducer operating at a nominal frequency of 50 kHz was used to drive an ultrasonic standing wave of 150 dB in pressure amplitude. The microsphere size distributions measured at the outlet were compared with the predictions of a theoretical model. Experiments and theory show reasonable correspondence. The theoretical model also indicates an optimal partitioning of the particle-laden and clean air inlet streams. PMID- 19682720 TI - Defining the role of CD46, CD80 and CD86 in mediating adenovirus type 3 fiber interactions with host cells. AB - Attachment of human adenoviruses (Ads) to host cells is mediated by the interaction of the fiber protein of the capsid with specific cell-surface molecules. For one of the species B adenoviruses, Ad3, the mechanism of binding to cells remains to be defined. Several previous reports have proposed CD46, CD80 or CD86 as possible Ad3 fiber attachment molecules. In this study, CD80 and CD86 were not found to mediate Ad3 fiber binding or Ad3-EGFP transduction of cells. Low levels of Ad3-EGFP transduction of a CHO cell line expressing relatively high levels of CD46 were detected which might suggest a role for CD46 in facilitating Ad3: cell interactions, in the absence of other attachment molecules. Anti-CD46 antibodies and siRNAs had almost no effect on Ad3 fiber binding or Ad3-EGFP transduction of HeLa cells. However, treatment of A549 cells with CD46 siRNA resulted in some decrease of transduction with Ad3-EGFP. PMID- 19682721 TI - Gel-forming exopolysaccharides explain basic differences between structures of aerobic sludge granules and floccular sludges. AB - The sol-gel transition of extracellular polymeric substances (EPS) derived from sludge flocs and granules is investigated in order to explain basic differences between the two aggregates. A reversible, pH dependent sol-gel transition was observed at pH 9.0-12.0 in EPS extracted from granules. At pH <9 granule EPS existed as a strong gel, indicating that their EPS exist in a gel state at normal operating pH of a wastewater treatment system (i.e. 6.0-8.5). This characteristic transition from solution to strong gel was not observed in any of the EPS samples derived from floccular sludges. A transition to a weak gel was however, observed at pH 4.0-5.0. Enriched exopolysaccharides from the granular EPS exhibited rheological behaviour analogous to the granules and the granule EPS. The critical overlap concentration (c*) of the exopolysaccharide concentrate was 0.33% w/w, similar to the c* of other known bacterial exopolysaccharides. Additionally, the protein content was found to be not contributing to the storage modulus of granule EPS gels. These factors suggest that exopolysaccharides or glycosides were the gelling agent in aerobic sludge granules. Given that EPS derived from aerobic sludge granules and flocs are distinguished by such a sol-strong gel transition, these exopolysaccharides therefore likely play an important role in granulation. PMID- 19682722 TI - Arsenic concentration in porewater of an alkaline coal ash disposal site: Roles of siderite precipitation/dissolution and soil cover. AB - The geochemical behavior of As in porewaters of an alkaline coal ash disposal site was investigated using multilevel samplers. The disposal site was in operation from 1983 until 1994 and was covered with 0.3-0.5m thick soils in 2001 when this study was initiated. Sequential extraction analyses and batch leaching experiments were also performed using the coal ash samples collected from the disposal site. The results suggest the important roles of siderite (FeCO(3)) precipitation/dissolution and soil cover, which have been ignored previously. Arsenic levels in the porewater were very low (average of 10microgL(-1)) when the site was covered with soil due to coprecipitation with siderite. The soil cover enabled the creation of anoxic conditions, which raised the Fe concentration by the reductive dissolution of Fe-(hydr)oxides. Because of the high alkalinity generated from the alkaline coal ash, even a small increase in the Fe concentration (0.66mgL(-1) on average) could cause siderite precipitation. When the soil cover was removed, however, an oxidizing condition was created and triggered the precipitation of dissolved Fe as (hydr)oxides. As a result, the dissolution of previously precipitated As-rich siderite caused higher As concentration in the porewater (average of 345microgL(-1)). PMID- 19682723 TI - Probabilistic environmental risk characterization of pharmaceuticals in sewage treatment plant discharges. AB - The occurrence of pharmaceuticals in different water bodies and the findings of effects on aquatic organisms in ecotoxicity tests have raised concerns about environmental risks of pharmaceuticals in receiving waters. Due to the fact that the amount of ecotoxicological studies has increased significantly during the last decade, probabilistic approaches for risk characterization of these compounds may be feasible. This approach was evaluated by applying it to 22 human used pharmaceuticals covering both pharmaceuticals with a high volume and high ecotoxicity, using ecotoxicological effect data from laboratory studies and comparing these to monitoring data on the effluents from sewage treatment plants in Europe and pharmaceutical sales quantities. We found that for 19 of the 22 selected pharmaceuticals the existing data were sufficient for probabilistic risk characterizations. The subsequently modeled ratios between monitored concentrations and low-effect concentrations were mostly above a factor of 100. Compared to the current paradigm for EU environmental risk assessment where a safety factor of 10 or 100 might have been used it seems that for the modeled compounds there's a low environmental risk. However, similarly calculated ratios for five pharmaceuticals (propranolol, ibuprofen, furosemide, ofloxacin, and ciprofloxacin) were below 100, while ibuprofen and ciprofloxacin are considered to be of high concern due to lack of ecotoxicity studies. This paper shows that by applying probabilistic approaches, existing data can be used to execute a comprehensive study on probability of impacts, thereby contributing to a more comprehensive environmental risk assessment of pharmaceuticals. PMID- 19682724 TI - The toxic effects of ionic liquids on the activities of acetylcholinesterase and cellulase in earthworms. AB - The earthworm Eisenia foetida was exposed to different concentrations of imidazolium ionic liquids with varying chain lengths according to the method of OECD [OECD, 1984. (The Current Organization of Economic and Cooperative Development Acute Earthworm Toxicity Test) Guidelines for the Testing of Chemicals, No. 207. Earthworm Acute Toxicity Tests]. The acute and subchronic toxic effects of [C(8)mim]Br on the activities of acetylcholinesterase (AChE) and cellulase in earthworms were determined under an artificial soil condition. Using filter paper contact tests, the 48 h-LC(50) values of [C(4)mim]Br, [C(6)mim]Br, [C(8)mim]Br, [C(10)mim]Br and [C(12)mim]Br on the earthworm were 73.33, 28.25, 2.69, 0.37 and 0.02 microg cm(-2), respectively. The 7 d-LC(50) of [C(8)mim]Br was 206.8 mg kg(-1) artificial soil (dry weight) and the 14 d-LC(50) was 159.4 mg kg(-1) artificial soil (dry weight), under the condition of artificial soil. After 1 d and 3 d of acute exposure, the activity of AChE was markedly inhibited when compared to the control, while it was increased at 7d. The cellulase activity was elevated significantly in the treatment groups of 20-160 mg kg(-1) after 3 and 7d of acute exposure. The activity of cellulase was also promoted under the subchronic exposure condition in the 10 and 20 mg kg(-1) groups. The experimental results suggest that [C(8)mim]Br may interfere with the nervous function of the earthworms and increase their cellulase activity. These results indicate that [C(8)mim]Br-exposure can affect the metabolized enzyme activity of earthworms at low concentrations and can even cause worm death at high doses, both of which have potential impacts on the soil environment. PMID- 19682726 TI - WITHDRAWN: Reply to Comment by Sierra Rayne and Kaya Forest on "Ab initio study of the structural, electronic, and thermodynamic properties of linear perfluorooctane sulfonate (PFOS) and its branched isomers" AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19682725 TI - Patterns and concentration levels of polybrominated diphenyl ethers (PBDEs) in placental tissue of women in Denmark. AB - The levels and congener patterns of PBDEs were investigated in human placental samples in Denmark. The median concentrations of sigmaPBDE(tri-hepta) and BDE-209 in the 50 samples were 1.22 and 1.14 ng g(-1) lw, respectively, with the total sum ranging from 0.51 to 17.1 ng g(-1) lw, which is similar to previous placental studies. The PBDE content in placental tissue was dominated by BDE-209, which accounted for approximately 50% of the total amount of PBDEs. BDE-47, -99, and 153 were detected in all samples. Approximately equal amounts of BDE-47 and BDE 153 were observed in the placental tissue, which is in agreement with previous European studies of human serum. Principal Component Analysis (PCA) was performed to analyze congener patterns within and between mothers. The loading plot showed groupings of the measured PBDE variables in three groups, representative of Penta , Octa- and Deca-BDE technical mixtures. Congeners representing the individual technical mixtures were close to orthogonal or inversely correlated, indicating variation in the congener patterns of internal exposure corresponding to the patterns of technical mixtures used in products. Visualisation of the participant objects according to body mass index (BMI), revealed inherent congener patterns (19% X-variance) showing increased frequency for participants within the highest BMI group to have elevated concentrations of BDE-209 in the placental tissue. PMID- 19682727 TI - Removal of personal care compounds from sewage sludge in reed bed container (lysimeter) studies--effects of macrophytes. AB - Sludge reed beds have been used for dewatering (draining and evapotranspiration) and mineralisation of sludge in Europe since 1988. Although reed beds are considered as a low cost and low contamination method in reducing volume, breaking down organic matter and increasing the density of sludge, it is not yet clear whether this enhanced biological treatment is suitable for degradation of organic micro-pollutants such as personal care products. Within this project the effect of biological sludge treatment in a reed bed on reducing the concentrations of the fragrances HHCB, AHTN, OTNE was studied as on the bactericide Triclosan. Additionally, the capacity of different macrophytes species to affect the treatment process was examined. Three different macrophyte species were compared: bulrush (Typha latifolia), reed (Phragmites australis) and reed canary grass (Phalaris arundinacea). They were planted into containers (lysimeters) with a size of 1 m x 1 m x 1 m which were filled with 20 cm gravel at the bottom and 50 cm sludge on top, into which the macrophytes were planted. During the twelve months experiment reduction of 20-30% for HHCB and AHTN, 70% for Triclosan and 70% for OTNE were determined under environmental conditions. The reduction is most likely due to degradation, since volatilization, uptake into plants and leaching are insignificant. No difference between the containers with different macrophyte species or the unplanted containers was observed. Considering the usual operation time of 10 years for reed beds, an assessment was made for the whole life time. PMID- 19682728 TI - Mobile phone radiation inhibits Vigna radiata (mung bean) root growth by inducing oxidative stress. AB - During the last couple of decades, there has been a tremendous increase in the use of cell phones. It has significantly added to the rapidly increasing EMF smog, an unprecedented type of pollution consisting of radiation in the environment, thereby prompting the scientists to study the effects on humans. However, not many studies have been conducted to explore the effects of cell phone EMFr on growth and biochemical changes in plants. We investigated whether EMFr from cell phones inhibit growth of Vigna radiata (mung bean) through induction of conventional stress responses. Effects of cell phone EMFr (power density: 8.55 microW cm(-2); 900 MHz band width; for 1/2, 1, 2, and 4 h) were determined by measuring the generation of reactive oxygen species (ROS) in terms of malondialdehyde and hydrogen peroxide (H(2)O(2)) content, root oxidizability and changes in levels of antioxidant enzymes. Our results showed that cell phone EMFr significantly inhibited the germination (at > or =2 h), and radicle and plumule growths (> or =1 h) in mung bean in a time-dependent manner. Further, cell phone EMFr enhanced MDA content (indicating lipid peroxidation), and increased H(2)O(2) accumulation and root oxidizability in mung bean roots, thereby inducing oxidative stress and cellular damage. In response to EMFr, there was a significant upregulation in the activities of scavenging enzymes, such as superoxide dismutases, ascorbate peroxidases, guaiacol peroxidases, catalases and glutathione reductases, in mung bean roots. The study concluded that cell phone EMFr inhibit root growth of mung bean by inducing ROS-generated oxidative stress despite increased activities of antioxidant enzymes. PMID- 19682729 TI - Performance of two relatively new quantitative D-dimer assays (Innovance D-dimer and AxSYM D-dimer) for the exclusion of deep vein thrombosis. AB - INTRODUCTION: D-dimer assays are now widely used as the first-line test in the diagnostic algorithm of suspected deep vein thrombosis (DVT). The aim of this study was to evaluate the performance of two relatively new quantitative D-Dimer assays (Innovance and AxSYM) by comparison with a clinical gold standard. PATIENTS AND METHODS: 311 samples from outpatients with clinical suspicion of DVT, included in a prospective management study, was analysed (prevalence of DVT 23%). The diagnostic workup included estimation of pre-test probability, D-dimer determination, objective imaging as well as 3 month clinical follow up of negative patients. RESULTS: No significant differences were seen in sensitivity and negative predictive values between Innovance, AxSYM and the reference assays. The area under the ROC curve was slightly lower for the AxSYM assay and the correlation to the reference assays was only moderate (r < 0.8) whereas the agreement with the Vidas assay was near excellent (kappa = 0.8). The Innovance assay reached the highest AUC, showed a strong correlation with the reference assays (r > or = 0.9) and a good agreement with the Vidas assay (kappa = 0.76). In combination with a low pre-test probability score the Innovance assay reached a NPV of 100% (95% CI, 92-100) and the AxSYM assay 98% (95% CI, 87-100). CONCLUSION: The Innovance and AxSYM assays show an overall good and comparable performance for the exclusion of DVT when compared to the established assays. Our results for the AxSYM assay indicate that the optimal cut-off value needs to be further evaluated. PMID- 19682731 TI - Sampling techniques for oviductal and uterine luminal fluid in cattle. AB - Analysis of luminal fluid microenvironments in the reproductive tract is pivotal to elucidate embryo-maternal signaling mechanisms responsible for successful reproduction in mammals, including cattle. Besides facilitating production of an optimized medium for in vitro fertilization and embryo culture in assisted reproductive technologies, screening of luminal fluid constituents in the oviduct and uterus could also provide critical information for elucidation of mechanisms underlying developmental programming. A key issue in this type of research is the sampling of luminal fluids. In this review we discuss the sampling techniques available for bovine species, including a recent in situ technique developed with the Ghent device, which allows rapid recovery of measurable amounts of pure uterine luminal fluid with minimal disturbance to the donor animal. PMID- 19682730 TI - Identification of selective inhibitors of cancer stem cells by high-throughput screening. AB - Screens for agents that specifically kill epithelial cancer stem cells (CSCs) have not been possible due to the rarity of these cells within tumor cell populations and their relative instability in culture. We describe here an approach to screening for agents with epithelial CSC-specific toxicity. We implemented this method in a chemical screen and discovered compounds showing selective toxicity for breast CSCs. One compound, salinomycin, reduces the proportion of CSCs by >100-fold relative to paclitaxel, a commonly used breast cancer chemotherapeutic drug. Treatment of mice with salinomycin inhibits mammary tumor growth in vivo and induces increased epithelial differentiation of tumor cells. In addition, global gene expression analyses show that salinomycin treatment results in the loss of expression of breast CSC genes previously identified by analyses of breast tissues isolated directly from patients. This study demonstrates the ability to identify agents with specific toxicity for epithelial CSCs. PMID- 19682733 TI - Molecules involved in sperm-oviduct adhesion and release. AB - In mammals, sperm ascension within the female reproductive tract involves a transient adhesion to the caudal isthmus of the oviduct. Sperm adhesion to this specialized region, which is termed the "oviductal reservoir", extends the sperm fertile life span by delaying capacitation until, around ovulation, specific signals induce sperm release. In vivo and in vitro studies demonstrated that carbohydrates on the oviductal cell apical membranes and lectin-like molecules on the rostral sperm surface are involved in adhesion in a species-specific way. In this respect, the most intensely studied species are pigs and cattle. On the other hand, less is known about molecules involved in sperm release. Direct evidence that molecules present in the oviductal fluid trigger the release of sperm bound to in vitro cultured oviductal epithelium has been provided only in cattle. However, the identity of sperm and/or oviductal molecules that respond to these releasing signals is still unknown. The comprehension of molecular mechanisms underlying sperm-oviduct interaction may advance our understanding of the behavior of sperm within the female reproductive tract and provide new tools for sperm selection, extension of fertile life and modulation of capacitation in the field of reproductive biotechnologies. The aim of the present paper is to review the available knowledge on molecules involved in sperm selection, storage and release from the oviductal reservoir. PMID- 19682732 TI - Endoscopic approaches to manage in vitro and in vivo embryo development: use of the bovine oviduct. AB - The oviduct plays a major part in different reproductive processes providing the microenvironment for numerous steps in early embryogenesis. Consequently, there is a growing demand to perform comparative studies focusing on causal mechanisms related to embryo development within its environment including complex and holistic strategies. However, the routine flushing and transfer procedure of bovine embryos is limited to the morula and blastocyst stage. Additionally, the use of in vitro production of bovine embryos provides access to an extra amount of embryos at various stages. But the quality of these embryos does not reflect the quality of its ex vivo counterparts. For two decades our own studies have focused on use of the oviductal environment of different species to optimize early embryo development for different purposes. The current article briefly highlights some main characteristics of the fallopian tube and reviews the endoscopic approach to access the fallopian tube using the stepwise minimal invasive technique established in different species. PMID- 19682734 TI - In vitro cytotoxicity evaluation of elemental ions released from different prosthodontic materials. AB - OBJECTIVES: This study investigated the cytotoxicity of elemental ions contained in four fixed prosthodontic materials (gold, nickel-chromium, stainless-steel alloys and CAD-CAM ceramics). MATERIALS AND METHODS: According to the determination of elements released from prosthodontic materials by using inductively coupled plasma mass spectroscopy, similar amounts of elements Pd, Ag, Zn, Cu, Ni, Cr, Mo, Be, Fe, Al, and K were prepared as salt solutions. Wells with a tenfold higher concentration of the tested elements were used as positive controls, while a well without any tested element was used as a negative control. These salt solutions were tested for cytotoxicity by culturing mouse L-929 fibroblasts in the salt solutions for a 7-day period of incubation. Then, the percentage of viable cells for each element was measured using trypan blue exclusion assay. The data (n=5) were statistically analyzed by ANOVA/Tukey test (p<0.05). RESULTS: The results showed a statistically significant difference for the cytotoxic effect of the tested elements salt solutions. For the released element concentrations the lowest percentage of viable cells (mean+/-SD) was evident with Zn, Cu or Ni indicating that they are the highly toxic elements. Be and Ag were found to be intermediate in cytotoxic effect. Fe, Cr, Mo, Al, Pd or K were found to be the least cytotoxic elements. SIGNIFICANCE: Zn and Cu released from gold alloys, and Ni released from nickel-chromium alloys, which are commonly used as fixed prosthodontic restorations, show evidence of a high cytotoxic effect on fibroblast cell cultures. PMID- 19682735 TI - Non-invasive ventilation after extubation in hypercapnic patients with chronic respiratory disorders: randomised controlled trial. AB - BACKGROUND: Non-invasive ventilation can prevent respiratory failure after extubation in individuals at increased risk of this complication, and enhanced survival in patients with hypercapnia has been recorded. We aimed to assess prospectively the effectiveness of non-invasive ventilation after extubation in patients with hypercapnia and as rescue therapy when respiratory failure develops. METHODS: We undertook a randomised controlled trial in three intensive care units in Spain. We enrolled 106 mechanically ventilated patients with chronic respiratory disorders and hypercapnia after a successful spontaneous breathing trial. We randomly allocated participants by computer to receive after extubation either non-invasive ventilation for 24 h (n=54) or conventional oxygen treatment (n=52). The primary endpoint was avoidance of respiratory failure within 72 h after extubation. Analysis was by intention to treat. This trial is registered with clinicaltrials.gov, identifier NCT00539708. FINDINGS: Respiratory failure after extubation was less frequent in patients assigned non-invasive ventilation than in those allocated conventional oxygen therapy (8 [15%] vs 25 [48%]; odds ratio 5.32 [95% CI 2.11-13.46]; p<0.0001). In patients with respiratory failure, non-invasive ventilation as rescue therapy avoided reintubation in 17 of 27 patients. Non-invasive ventilation was independently associated with a lower risk of respiratory failure after extubation (adjusted odds ratio 0.17 [95% CI 0.06-0.44]; p<0.0001). 90-day mortality was lower in patients assigned non-invasive ventilation than in those allocated conventional oxygen (p=0.0146). INTERPRETATION: Early non-invasive ventilation after extubation diminished risk of respiratory failure and lowered 90-day mortality in patients with hypercapnia during a spontaneous breathing trial. Routine implementation of this strategy for management of mechanically ventilated patients with chronic respiratory disorders is advisable. FUNDING: IDIBAPS, CibeRes, Fondo de Investigaciones Sanitarias, European Respiratory Society. PMID- 19682736 TI - Leaving invasive ventilation behind. PMID- 19682737 TI - Candidate biomarker discovery in plasma of juvenile cod (Gadus morhua) exposed to crude North Sea oil, alkyl phenols and polycyclic aromatic hydrocarbons (PAHs). AB - In this study we have investigated protein changes in plasma of juvenile Atlantic cod (Gadus morhua) induced by crude North Sea oil and North Sea oil spiked with alkyl phenols and polycyclic aromatic hydrocarbons, a surrogate produced water composition. Using a proteomic approach, we identified 137 differentially expressed proteins at different levels of crude oil exposure. Many of the induced protein changes occurred at low levels of exposure. The results obtained with protein expression profiles after exposure to oil and surrogate produced water indicate effects on fibrinolysis and the complement cascade, the immune system, fertility-linked proteins, bone resorption, fatty acid metabolism as well as increased oxidative stress, impaired cell mobility and increased levels of proteins associated with apoptosis. Although the number of individuals and samples in this study is limited within each treatment group, the protein changes observed in this study represent a first screening for potential biomarker candidates in cod plasma reflecting potential effects of crude oil and produced water exposure on fish. PMID- 19682738 TI - Rising water temperatures, reproduction and recruitment of an invasive oyster, Crassostrea gigas, on the French Atlantic coast. AB - The recent appearance and invasion of feral oysters (Crassostrea gigas) along the northern European Atlantic coast, underscores the necessity to investigate the relationship between environmental variables, reproductive physiology, larval development and recruitment. We studied these relationships at both high (HT) and intermediate (IT) - turbidity sites, through historical data on water temperatures, multi-parameter environmental probes, histological analyses, and field collections of planktonic larvae and settled post-larvae in 2005 and 2006. A progressive warming trend was observed, especially since 1995, when oyster proliferation first became severe. Threshold temperatures for oocyte growth, larval development and settlement were achieved in both 2005 and 2006. The HT site showed greater numbers of larvae and post-larvae than the IT site for both years, with the highest numbers of post-larvae observed at both sites during the warmer summer of 2006. These results suggest that increased temperatures in northern European waters allow successful reproduction, larval development, and recruitment of C. gigas. High turbidity conditions further enhance this success. PMID- 19682739 TI - The use of green fluorescence gene (GFP)-modified rabbit mesenchymal stem cells (rMSCs) co-cultured with chondrocytes in hydrogel constructs to reveal the chondrogenesis of MSCs. AB - This study was conducted to reveal the chondrogenesis of mesenchymal stem cells that had been genetically modified with the green fluorescence protein (GFP) gene and then co-cultured with chondrocytes in vitro and in vivo. Subsequent mixing of chondrocytes in the hydrogel constructs induced increased chondrogenic differentiation of the transfected hMSCs. The proliferation and differentiation of MSCs that were transfected with the GFP gene and co-cultured with chondrocytes (1:1 and 1:3) or chondrocytes alone were evaluated by a live/dead assay, MTT assay, GAG & DNA assay, RT-PCR, real time-PCR, and histological and immunochemical analysis in vitro and in vivo. Real-time PCR revealed that the expression of aggrecan and COMP by genetically modified hMSCs co-cultured with chondrocytes was 2 or 3 times greater than that of genetically modified MSCs alone. Moreover, the expression of collagen type II was more than 3.5 times greater than that of genetically modified MSCs alone. 3-D hydrogel constructs co cultured with chondrocytes and genetically modified MSCs showed a significantly higher number of specific lacunae phenotypes at the end of the 4 week study, regardless of whether they were co-cultured in the presence of chondrocytes. These findings indicate that co-culture with chondrocytes and genetically modified MSCs can be used to engineer well designed implants for the formation of neocartilage by transplanted genetically modified MSCs. PMID- 19682740 TI - Combined delivery of heme oxygenase-1 gene and fibroblast growth factor-2 protein for therapeutic angiogenesis. AB - Ectopic expression of heme oxygenase-1 (HO-1) in ischemic tissue protects the tissue from apoptosis and necrosis and promotes angiogenesis. However, apoptosis and necrosis will decrease HO-1 gene transfection efficacy. We hypothesized that fibroblast growth factor-2 (FGF2) would attenuate ischemic damage during the incipient period, improve HO-1 gene transfection and, in turn, enhance neovascularization. To test this hypothesis, we employed a mouse model of hindlimb ischemia and treated the mice with HO-1 gene therapy alone, FGF2 alone, or HO-1 gene therapy plus FGF2. As controls, a group of mice was left untreated. At 12h, prior to the expression of exogenously delivered HO-1, apoptosis was significantly reduced in mice treated with FGF2, either alone or in combination with HO-1 gene therapy. At 3 days, HO-1 expression was greater in mice that also received FGF2 than in mice treated with HO-1 gene therapy alone. The expression of angiogenic growth factors and angiogenesis was greater in mice treated with HO 1 gene therapy plus FGF2 than in mice treated with HO-1 gene therapy alone. These data indicate that FGF2 rescued muscle necrosis prior to the exogenous expression of HO-1 and enhanced HO-1 gene transfection in ischemic murine limbs. PMID- 19682741 TI - Dual modulation of CNS voltage-gated calcium channels by cannabinoids: Focus on CB1 receptor-independent effects. AB - The neuromodulatory effects of cannabinoids in the central nervous system have mainly been associated with G-protein coupled cannabinoid receptor (CB1R) mediated inhibition of voltage-gated calcium channels (VGCCs). Numerous studies show, however, that cannabinoids can also modulate VGCCs independent of CB1R activation. Nevertheless, despite the fact that endocannabinoids have a nearly equal efficacy for direct and CB1R-mediated effects on VGCC, the role of the direct cannabinoid-VGCC interaction has been largely underestimated. In this review, we summarize recent studies on the modulation of different types of VGCCs by cannabinoids, highlight the evidence for and implications of the CB1R independent modulation, and put forward the concept, that direct interaction of cannabinoids and VGCCs is as important in regulation of VGCCs function as the CB1R-mediated effects. PMID- 19682742 TI - How to improve the treatment outcome in chronic lymphocytic leukemia? PMID- 19682743 TI - Preclinical antileukemia activity of JNJ-26481585, a potent second-generation histone deacetylase inhibitor. AB - Histone deacetylase (HDAC) inhibitors have been shown to induce cell cycle arrest, terminal differentiation, and apoptosis in a broad spectrum of human tumors and animal xenograft models. JNJ-26481585 is a hydroxamic acid derivative, second-generation pan-HDAC inhibitor that has demonstrated high potency in preclinical studies. In the current study, we demonstrated that JNJ-26481585 has antileukemia and molecular activity in leukemia cell lines and primary human leukemia cells. We also observed a synergistic effect between treatment with decitabine and JNJ-26481585. In conclusion, JNJ-26481585 is a potent second generation pan-HDAC inhibitor with activity in human leukemia, and it is currently in clinical development. PMID- 19682744 TI - Inter-tissue distribution and evaluation of potential toxicity of PCBs in Baltic cod (Gadus morhua L.). AB - PCBs measured in the muscle, liver, and gonads of cod collected from two locations in the southern Baltic were evaluated as total (summation operator PCBs), indicator (summation operator PCB(7)), and dioxin-like (summation operator DL-PCBs) concentrations. Gender, tissue lipid content, and collection site were important determinants of PCB levels. Irrespective of collection site, lipid normalized summation operator PCBs, summation operator PCB(7), and summation operator DL-PCBs in females were the lowest in the muscle, and of similar levels in liver and gonads. In males the concentrations had a decreasing order of liver >muscle >gonads. Despite the gender differences in tissue levels, the profiles were fairly similar in both genders and sites. Significant relationships were found for both summation operator DL-PCBs and summation operator TEQ(DL-PCB) and concentrations of summation operator PCBs and summation operator PCB(7). The levels of summation operator TEQ(DL-PCB) in the livers of both genders and in female gonads, in addition to the total PCBs load in these organs, raise concern and suggest that the species can suffer from PCBs toxic effects. PMID- 19682745 TI - Epidermal nevus syndrome: an unusual cerebellar involvement. AB - The epidermal nevus syndrome is characterized by several developmental anomalies associated with an epidermal nevus. In addition to the skin, other organs commonly affected include the brain, eyes and musculoskeletal system. We report here on a 24-year-old woman with this syndrome who presented with hemifacial hypertrophy, hearing abnormalities, arrhythmia and an unusual infratentorial brain involvement. PMID- 19682746 TI - Life cycle assessments of municipal solid waste management systems: a comparative analysis of selected peer-reviewed literature. AB - Life cycle assessment (LCA) is a popular tool used to evaluate the environmental performance of municipal solid waste (MSW) management systems. Although reviews of LCAs of MSW have been undertaken to assess the validity of the 'waste hierarchy,' a recent review of the goal, scope and results of LCAs of mixed material MSW management systems has yet to be performed. This paper is a comparative analysis of 20 process-based LCAs of MSW published between 2002 and 2008 in a total of 11 English-language peer-reviewed journals. It quantifies the methodological transparency of the studies and the frequency of use of particular system boundaries, types of data sources, environmental impact categories, impact weightings, economic valuations, sensitivity analyses, and LCA computer models. Net energy use (NEU), global warming potential (GWP), and acidification potential (AP) values for various types of MSW management systems are also compared using statistical indicators. The reviewed LCAs differ substantially in their system boundaries. Half or more of the LCAs either do not mention or are unclear in whether or not life cycle emissions from energy inputs or capital equipment are included in the calculation of results. Only four impact categories are common to more than half of the reviewed LCAs. The human and ecological toxicity impact categories are much less common than global warming potential, acidification, and eutrophication. A financial life cycle costing is present in eight of the reviewed LCAs, while an economic valuation of the environmental impacts is observed in five. Explicit sensitivity analyses are present in 4/20 of the studies, although many more LCAs evaluate the effects of varying model parameters by increasing the number of waste management scenarios. There is no consensus on whether or not to use the marginal or average source of electricity in calculating environmental impacts. Eight out of the 20 do not mention this source while the remaining LCAs are evenly split between the marginal and average electricity source. One quarter of the reviewed LCAs supply weighted results for the overall environmental performance of MSW management scenarios. All but one of these concurred with the 'hierarchy of waste' that the environmental performance of landfilling is lower than that of all the other treatment methods, and that thermal treatments are inferior to recycling. The comparative analyses of the NEU, GWP and AP results are based on 37, 45, and 42 MSW management scenarios, respectively. As measures of statistical dispersion, the interquartile ranges of the NEU, GWP and AP values are lowest for the landfilling (AP, NEU) and thermal treatment (GWP) scenarios. The results of the statistical analysis of the NEU, AP and GWP values appear to indicate that thermal treatment scenarios have a better environmental performance than landfilling, while the results for mixed treatment scenarios are less obvious. A comparison of the relative environmental performances of MSW treatment scenario types within each study did not provide a clear confirmation or repudiation of the waste hierarchy. This paper concludes that many recently published LCAs do not ensure that the methodological assumptions are made clear to the reader. Lack of transparency makes the results difficult to interpret, and hampers meaningful comparisons between the LCA results. A convergence in the adoption of particular assumptions that are more representative of MSW management systems would facilitate the comparison of the results. PMID- 19682747 TI - Experimental history: swinging pendulums and melting shellac. AB - Four hundred years ago Galileo Galilei aimed a telescope at the sky. He revolutionized astronomy. Equally revolutionary were his experiments in physics. Unlike his astronomical observations the experiments remain difficult to understand and replicate even today. Two centuries after Galileo, Augustin Coulomb demonstrated experimentally the law of electrostatic force. It has never been successfully replicated. Yet both Galileo and Coulomb were exquisite experimentalists. The fact is that revolutionary experiments in physics are never finished. They are open for investigation for generations to come. PMID- 19682748 TI - Leishmania major parasites induced macrophage tolerance: implication of MAPK and NF-kappaB pathways. AB - During infection pathogens elicit early inflammatory events and modulate mediators of the host inflammatory response. We show in the present study that the two stages of the Leishmania parasite differentially modulate the production of inflammatory cytokines with amastigotes inducing cytokine production but not promastigotes. However once they infect macrophages both stages were able to induce a state of tolerance characterized by the inhibition of TNFalpha production in response to a subsequent contact with the parasite. This effect was not due to the action of soluble deactivating cytokines released in the supernatant, but likely reflect direct cell-parasite interactions. Moreover, we show that parasite viability is required for macrophage tolerisation. Cross stimulation experiments using two stimuli namely Leishmania and LPS, demonstrated a hierarchy of signalling with LPS mediating abrogation of Leishmania tolerance but not vice versa. Indeed, while LPS was able to overcome Leishmania induced tolerance LPS induced cell tolerance was refractory to subsequent Leishmania stimulation. This state of tolerance correlates with the hypo responsiveness of MAPK transduction pathways and defective activation of NF-kappaB transcription factor. PMID- 19682749 TI - Canadian Network for Mood and Anxiety Treatments (CANMAT) clinical guidelines for the management of major depressive disorder in adults. II. Psychotherapy alone or in combination with antidepressant medication. AB - BACKGROUND: In 2001, the Canadian Psychiatric Association and the Canadian Network for Mood and Anxiety Treatments (CANMAT) partnered to produce evidence based clinical guidelines for the treatment of depressive disorders. A revision of these guidelines was undertaken by CANMAT in 2008-2009 to reflect advances in the field. This article, one of five in the series, reviews new studies of psychotherapy in the acute and maintenance phase of MDD, including computer-based and telephone-delivered psychotherapy. METHODS: The CANMAT guidelines are based on a question-answer format to enhance accessibility to clinicians. Evidence based responses are based on updated systematic reviews of the literature and recommendations are graded according to the Level of Evidence, using pre-defined criteria. Lines of Treatment are identified based on criteria that included evidence and expert clinical support. RESULTS: Cognitive-Behavioural Therapy (CBT) and Interpersonal Therapy (IPT) continue to have the most evidence for efficacy, both in acute and maintenance phases of MDD, and have been studied in combination with antidepressants. CBT is well studied in conjunction with computer-delivered methods and bibliotherapy. Behavioural Activation and Cognitive-Behavioural Analysis System of Psychotherapy have significant evidence, but need replication. Newer psychotherapies including Acceptance and Commitment Therapy, Motivational Interviewing, and Mindfulness-Based Cognitive Therapy do not yet have significant evidence as acute treatments; nor does psychodynamic therapy. LIMITATIONS: Although many forms of psychotherapy have been studied, relatively few types have been evaluated for MDD in randomized controlled trials. Evidence about the combination of different types of psychotherapy and antidepressant medication is also limited despite widespread use of these therapies concomitantly. CONCLUSIONS: CBT and IPT are the only first-line treatment recommendations for acute MDD and remain highly recommended for maintenance. Both computer-based and telephone-delivered psychotherapy--primarily studied with CBT and IPT--are useful second-line recommendations. Where feasible, combined antidepressant and CBT or IPT are recommended as first-line treatments for acute MDD. PMID- 19682750 TI - Canadian Network for Mood and Anxiety Treatments (CANMAT) clinical guidelines for the management of major depressive disorder in adults. Introduction. PMID- 19682751 TI - Group A streptococcal puerperal sepsis: initial characterization of virulence factors in association with clinical parameters. AB - Group A beta-hemolytic streptococcus (GAS) is an uncommon but potentially fatal source of postpartum infection. Pathogenesis in invasive GAS infections has been linked to bacterial virulence factors. In this study, we sought to provide an initial description of potential virulence factors in association with puerperal morbidity by virtue of specific M-protein type antigens. Women with confirmed GAS puerperal infection in the Salt Lake City region were prospectively identified over a 6-year interval (1991-1997). From this cohort, GAS isolates were analyzed with respect to M-serotype and presence of genes encoding the Streptococcal Pyogenic Exotoxins A and B (SPE-A and SPE-B). Bacterial isolates from 18 subjects with GAS puerperal infection underwent M-serotyping and PCR-based genotyping for the speA and speB genes. Among these, 8/18 subjects manifest criteria of severe disease. All 18 isolate strains expressed speB; 6/18 isolates expressed speA. Of the M-serotypes, 8/8 severe disease isolates expressed M-types 1 (N=3) or 28 (N=5). Pulse-field gel electrophoresis did not indicate an outbreak strain among similar isolates. We conclude that in this initial characterization, morbidity among women with GAS puerperal infection is associated with M-types 1 and 28, but not speB genotype. PMID- 19682752 TI - Classical and non-classical Major Histocompatibility Complex class I gene expression in in vitro derived bovine embryos. AB - The role of the Major Histocompatibility Complex class I (MHC-I) genes in human and mouse preimplantation embryo development has received considerable attention. In contrast, information concerning the role of these genes in bovine embryo development is limited. The objective of the current study was to characterize the expression pattern of MHC-I genes during preimplantation embryo development in cattle. To this end, bovine oocytes were harvested from slaughterhouse ovaries, matured, fertilized and cultured in vitro. Samples were collected at immature and mature oocyte, presumptive zygote, 2-4-cell embryo, 8-16-cell embryo, morula, blastocyst and hatched blastocyst stages of development. MHC-I expression was detected using quantitative real-time-PCR, cDNA sequencing, whole mount immunocytochemistry and Western blotting. We report classical and non classical MHC-I mRNA expression in bovine oocytes and developing embryos. Furthermore, we report that the pattern of MHC-I mRNA expression across development was gene- and stage-specific. PMID- 19682753 TI - Altered distribution of NK and NKT cells in follicular fluid is associated with IVF outcome. AB - The aim of this study was to investigate differences in the relative distributions of subsets of natural killer (NK) cells, including immunoregulatory NK cells (CD56(+)CD16(-)), cytotoxic NK cells (CD56(+)CD16(+)), as well as total NK cells (CD56(+)CD3(-)), and NKT cells (CD56(+)CD3(+)) in peripheral blood and follicular fluid in subjects with successful or unsuccessful IVF treatment. The immunoregulatory NK cell population in follicular fluid of women who failed to achieve pregnancy after IVF treatment was significantly decreased compared to women who became pregnant after IVF. Conversely, the NKT cell population in the follicular fluid of women with unsuccessful treatment was significantly elevated compared with those with successful IVF. Understanding the changes in the distribution of NK and NKT cell populations in follicular fluid might serve as the basis for a more detailed study to determine whether NK cell parameters have prognostic value in guiding the selection of individual ova for use in IVF procedures. PMID- 19682754 TI - Rational design of synthetic peptides to generate antibodies that recognize in situ CD11c(+) putative dendritic cells in horse lymph nodes. AB - A three-dimensional model of the alphaX I-domain of the horse integrin CD11c from dendritic cells provided information for selecting two segments of the primary structure for peptide synthesis. Peptide 1 contains 20 amino acids and peptide 2 has 17 amino acid residues. The first spans from position Thr229 to Arg248 of an alpha-helix segment of the structure, whereas peptide 2 goes from Asp158 to Phe174 and corresponds to an exposed segment of the loop considered to be the metal ion-dependent adhesion site. Murine polyclonal antisera against both peptides were generated and assayed in peripheral blood cell suspensions and in cryosections of horse lymph nodes. Only the serum against peptide 2 was capable of identifying cells in suspension and in situ by immunohistochemistry, some with evident dendritic morphology. Using this approach, an immunogenic epitope exposed in CD11c was identified in cells from horse lymph node in situ. PMID- 19682755 TI - Evaluation of humoral immune response in adaptive immunity in ALS patients during disease progression. AB - In ALS, evidence suggests immune reactions in disease pathogenesis. Although immunological changes point to adaptive immune response, whether humoral or cellular response dominates during disease course is unknown. The study aim was to evaluate humoral immune response in ALS patients during disease progression. Circulating immune complexes (CICs), IgG, and IgM in sera of ALS patients and matching controls were evaluated after each of two visits. Results showed significantly elevated CICs and IgG in ALS patient sera. CICs decreased to control levels at the second visit, yet IgG remained higher than controls. Serum IgM was within normal range. Results suggest a humoral immune response initiating adaptive immunity in ALS, however, cellular immune response needs verification. PMID- 19682757 TI - Relative importance of factors influencing the prevalence of lameness in Austrian cubicle loose-housed dairy cows. AB - The development of lameness is influenced by a number of different factors (housing, management, human-animal relationship and animal-related parameters). The aim of this study was to investigate the effects of the complex interactions of these aspects and to search for the relative importance of single factors. In 80 dairy herds of Austrian Simmental cows housed in cubicle loose housing in Upper and Lower Austria, risk factors for lameness, selected from the four factor groups housing, management, human-animal relationship and animal-related variables, were investigated during one farm visit in the autumn and winter months. To assess their relative importance, a multivariable analysis (regression trees) was calculated. The most important risk factor for lameness was the lying surface: straw bedding of at least 2 cm thickness or cow-comfort mats were associated with a lower percentage of lame cows. In case of insufficient quality of the lying surface, the next important parameter identified was the position of the neck rail: a neck rail diagonal greater than 1.94 m was associated with a lower percentage of lame cows. By contrast, on farms with high-quality lying surfaces, lameness prevalence was lower when at least parts of the alleys were constructed with solid floor and not slatted. Further variables associated with a low prevalence of lameness were a longer time span between calving and separation of the calf from the dam, the existence of an outside run, a lower percentage of fat cows, a greater space allowance, more cubicles than animals and a lower kerb height. In addition, further management factors such as the way in which heifers are integrated into the herd or management decisions taking into account the cows' welfare were related to less lameness. Human-animal relationship variables such as, for example, the behaviour and attitude of the stockpeople were explaining variables. In sum, important risk factors were found in all factor groups. Therefore it is necessary to optimise all those different aspects mentioned above to reduce the risk of lameness. PMID- 19682756 TI - Interaction of phase variation, host and pressure/gas composition: pneumococcal gene expression of PsaA, SpxB, Ply and LytA in simulated middle ear environments. AB - OBJECTIVE: Streptococcus pneumoniae, a leading cause of otitis media (OM), undergoes spontaneous intra-strain variations in colony morphology. Transparent (T) variants are more efficient in colonizing the nasopharynx while opaque (O) variants exhibit greater virulence during systemic infections. This study was intended to delineate the underlying molecular mechanisms by which the predominant S. pneumoniae variant efficiently infects the middle ear (ME) mucosa. METHODS: Human ME epithelial cells were preconditioned for 24h under one of the three gas/pressure conditions designed to simulate those for (1) normal ME (NME), (2) ME with Eustachian tube obstruction (ETO) and (3) ME with tympanostomy tube placement (TT), and then were incubated with ~ 10(7)CFU/ml of either T or O variants of S. pneumoniae (6A) for 3h. Relative expression levels of genes encoding virulence factors, PsaA (surface adhesion), SpxB (pyruvate oxidase), Ply (pneumolysin), and LytA (autolysin) were assessed separately in epithelium attached and supernatant bacteria 3h post infection using real-time PCR. RESULTS: Basal levels of the virulence molecules in inocula were comparable between two variants. However, relative expression levels of the gene transcripts were significantly induced in epithelium-attached T variants 3h after infection. Comparing with NME and TT conditions, ETO environment produced the largest effect on the differential expression of the virulence genes in the infected ME epithelial cells between T (induced) and O (suppressed) phenotypic pneumococci. CONCLUSIONS: T variant is a predominant phenotype responsible for the pathogenesis of pneumococcal OM. PMID- 19682758 TI - Dosimetric assessment of static and helical TomoTherapy in the clinical implementation of breast cancer treatments. AB - BACKGROUND AND PURPOSE: Investigation of the use of TomoTherapy and TomoDirect versus conventional radiotherapy for the treatment of post-operative breast carcinoma. This study concentrates on the evaluation of the planning protocol for the TomoTherapy and TomoDirect TPS, dose verification and the implementation of in vivo dosimetry. MATERIALS AND METHODS: Eight patients with different breast cancer indications (left/right tumor, axillary nodes involvement (N+)/no nodes (N0), tumorectomy/mastectomy) were enrolled. TomoTherapy, TomoDirect and conventional plans were generated for prone and supine positions leading to six or seven plans per patient. Dose prescription was 42Gy in 15 fractions over 3weeks. Dose verification of a TomoTherapy plan is performed using TLDs and EDR2 film inside a home-made wax breast phantom fixed on a rando-alderson phantom. In vivo dosimetry was performed with TLDs. RESULTS: It is possible to create clinically acceptable plans with TomoTherapy and TomoDirect. TLD calibration protocol with a water equivalent phantom is accurate. TLD verification with the phantom shows measured over calculated ratios within 2.2% (PTV). An overresponse of the TLDs was observed in the low dose regions (<0.1Gy). The film measurements show good agreement for high and low dose regions inside the phantom. A sharp gradient can be created to the thoracic wall. In vivo dosimetry with TLDs was clinically feasible. CONCLUSIONS: The TomoTherapy and TomoDirect modalities can deliver dose distributions which the radiotherapist judges to be equal to or better than conventional treatment of breast carcinoma according to the organ to be protected. PMID- 19682759 TI - Core needle biopsies for determination of the microenvironment in individual tumours for longitudinal radiobiological studies. AB - PURPOSE: We aimed to establish a core needle biopsy technique to investigate the impact on outcome of irradiation of the microenvironment in individual experimental tumours. METHODS: Nude mice bearing FaDu, UT-SCC-5, UT-SCC-14, and UT-SCC-15 tumours (n=67) were injected with pimonidazole hypoxia and Hoechst 33342 perfusion markers. One core needle biopsy was taken from the central part of the tumour under anaesthesia and the rest of the tumour was excised after marking the position of the needle. Relative hypoxic area (pHF), relative vascular area (RVA), fraction of perfused vessels (PF), and necrotic fraction (NF) were compared in the biopsies and the adjacent whole tumour sections. In a TCD(50) (dose to cure 50% of tumours) assay, 223 UT-SCC-5 tumours were irradiated with 30 fractions over 6weeks either with or without a core biopsy before the start of radiotherapy. RESULTS: The correlations between histological parameters measured in the biopsies and the adjacent tumour sections were dependent on the tumour line. All the four parameters showed weak although significant correlations only in UT-SCC-5. PF was the only parameter which showed a weak but significant correlation in all the four tumour lines. The needle biopsy procedure did not significantly impact on TCD(50) after fractionated irradiation in UT-SCC 5: 98Gy [92; 106] versus 105Gy [96; 117] (p=0.12). pHF, RVA, PF, and NF measured in the pre-treatment biopsy did not predict the outcome of fractionated irradiation within the UT-SCC-5 tumour line. CONCLUSION: A single pre-treatment core needle biopsy may provide valid results for parameters of the tumour micromilieu, however the accuracy is limited by significant intratumoural heterogeneity in the parameters and sampling error. The needle biopsy procedure does not significantly affect local tumour control rates after fractioned irradiation and may therefore be integrated for longitudinal studies in radiobiological experiments. Pre-treatment histological parameters measured in the biopsy did not correlate with the outcome of fractionated irradiation within the UT-SCC-5 tumour line. PMID- 19682760 TI - GTV spatial conformity between different delineation methods by 18FDG PET/CT and pathology in esophageal cancer. AB - PURPOSE: To find optimal threshold of length and GTV delineation for esophageal cancer using 18FDG PET/CT. MATERIALS AND METHODS: Sixteen patients with esophageal carcinoma underwent surgery. For each patient, six GTVs were defined. GTVCT was based on CT data alone. GTV20%, GTV40%, GTV2.5 and GTV40%M were generated by PET/CT, using SUVbgd + 20%(SUVmax(slice)--SUVbgd), SUVbgd + 40%(SUVmax(slice)--SUVbgd), 2.5 and 40%SUVmax(total) as thresholds. GTVpath was derived from pathology. Lengths of GTVs were recorded as LCT, L20%, L40%, L2.5, L40%M and Lpath, respectively. The former five GTVs/lengths were compared with GTVpath/Lpath by means of a conformity index CI/CI', which is the square of intersection of two GTVs/lengths divided by their product. RESULTS: Mean LCT, L20%, L40%, L2.5, L40%M and Lpath were 6.30 +/- 2.69, 5.55 +/- 2.48, 6.80 +/- 2.92, 6.65 +/- 2.66, 4.88 +/- 1.99 and 5.90 +/- 2.38 cm. Mean , , , and were 0.68 +/- 0.16, 0.84 +/- 0.17, 0.76 +/- 0.14, 0.78 +/- 0.15 and 0.80 +/- 0.11. and was significantly superior to (P < 0.05). Mean GTVCT, GTV20%, GTV40%, GTV2.5, GTV40%M and GTVpath were 29.16 +/- 18.56, 18.75 +/- 12.37, 12.52 +/- 8.08, 22.69 +/- 14.84, 9.18 +/- 5.96 and 28.16 +/- 17.02 cm3. Mean CIs increased significantly from CI40%&path(0.27 +/- 0.09) and CI'40% M&path (0.28 +/- 0.08) < CI'20% & path (0.52 +/- 0.16) and CI'2.5&path (0.52 +/- 0.20) < CICT&path(0.77 +/- 0.17). CONCLUSIONS: The SUVbgd + 20% (SUVmax(slice)--SUVbgd) method optimally estimated gross tumor length, but only reached an unsatisfactory CI for GTV. Due to possible motion factor enveloped in PET images and lack of histopathologic transverse reference, the information from both PET and CT should be referred to complementarily when delineating GTV. PMID- 19682761 TI - Adaptation and learning: characteristic time scales of performance dynamics. AB - A multiple time scales landscape model is presented that reveals structures of performance dynamics that were not resolved in the traditional power law analysis of motor learning. It shows the co-existence of separate processes during and between practice sessions that evolve in two independent dimensions characterized by time scales that differ by about an order of magnitude. Performance along the slow persistent dimension of learning improves often as much and sometimes more during rest (memory consolidation and/or insight generation processes) than during a practice session itself. In contrast, the process characterized by the fast, transient dimension of adaptation reverses direction between practice sessions, thereby significantly degrading performance at the beginning of the next practice session (warm-up decrement). The theoretical model fits qualitatively and quantitatively the data from Snoddy's [Snoddy, G. S. (1926). Learning and stability. Journal of Applied Psychology, 10, 1-36] classic learning study of mirror tracing and other averaged and individual data sets, and provides a new account of the processes of change in adaptation and learning. PMID- 19682762 TI - Analysis of the effect of nutritional factors on OTA and OTB biosynthesis and polyketide synthase gene expression in Aspergillus ochraceus. AB - The effect of a wide variety of nutritional based biotic factors on the production of both OTA and OTB biosynthesis in A. ochraceus was assessed. Different carbon sources including glucose, sucrose, maltose, galactose, xylose and glycerol appear to repress OTA production when the fungus is grown in OTA permissive PDY medium. In contrast lactose appears to induce OTA production, with the addition of lactose and galactose to the OTA restrictive PDC medium resulting in marked increases in OTA levels. The addition of lactose to MCB and PDY media considerably increases OTB production. The addition of both sucrose and galactose to MCB has similar yet less marked effects. Different nitrogen sources also affect OTA production with ammonium chloride significantly reducing OTA production, while organic nitrogen sources such as urea and amino acids including phenylalanine, lysine, glutamine and proline induce OTA production. The induction of otapksAo gene expression under these conditions correlates well with the levels of OTA produced under the same experimental conditions, suggesting that the observed effects appear to be modulated, at least in part, at the level of gene transcription. However while the levels of OTB produced in A. ochraceus also appear to be influenced by these nutritional based biotic factors, this appears to be regulated in a manner which is independent of otapksAo gene expression. PMID- 19682763 TI - Aldose reductase inhibitor counteracts the attenuated adhesion of human corneal epithelial cells induced by high glucose through modulation of MMP-10 expression. AB - AIMS: We investigated the preventive effect of aldose reductase inhibitor (ARI) on the adhesion of SV40-transformed human corneal epithelial cells (HCECs) exposed to high glucose, and the underlying mechanism focusing on the role of matrix metalloproteinase (MMP)-10. METHODS: HCECs were cultured in medium containing a normal (5.5 mM) or high (31.2 mM) concentration of D-glucose in the presence or absence of ARI, fidarestat. Cell attachment ability was evaluated by short-term adhesion assay. The levels of intracellular polyol were measured by liquid-gas chromatography. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR), and Western blotting were used to determine the expression levels. RESULTS: Decreased attachment activity and increased accumulation of polyol induced by exposure to high glucose were abrogated by ARI. Supply of recombinant MMP-10 decreased integrin alpha3beta1-expression and cell adhesion. The expression level of MMP-10 was enhanced at both protein and mRNA levels by exposure to high glucose, while that of integrin alpha3beta1 was decreased at the protein level, but remained unchanged at the mRNA level. These alterations in the expression levels of MMP-10 and integrin alpha3beta1 were normalized by ARI. CONCLUSIONS: ARI counteracts the decreased adhesion of HCECs induced by high glucose exposure, through the modulation of the expression of MMP-10. PMID- 19682764 TI - Pharmaceutical reforms: implications through comparisons of Korea and Japan. AB - This paper compares the process and results of the reform which confined doctors to prescribing and pharmacists to dispensing in both Korea and Japan from comparative and politico-economic perspectives. At the present time, several years since the reforms were implemented, a 'compulsory separation' is being established in Korea. The claims containing antibiotics against the total claims from the doctor's clinic dropped from 55.7% in 2000 to 29.6%, and the number of drugs per claim from 5.9 in 2000 to 4.2 in 2008. Japan selected an 'arbitrary separation'. Efforts to raise the rate of the 'separation' have increased the rate from 1% in 1974 to 57.2% in 2007, but nearly half of medical prescriptions are still being dispensed by doctors. Disparity in the two countries has been brought about by what follows: first, the president's political leadership caused a radical shift in the attitude of the bureaucratic group in Korea; second, in their confrontation with doctors the pharmacists' camp in Korea proved to hold political power stronger than that in Japan; third, intervention in policy of progressive civic groups in particular played a pivotal role in accomplishing the reform in Korea. PMID- 19682765 TI - Evaluating the mitochondrial timescale of human evolution. AB - Different methodologies and modes of calibration have produced disparate, sometimes irreconcilable, reconstructions of the evolutionary and demographic history of our species. We discuss how date estimates are affected by the choice of molecular data and methodology, and evaluate various mitochondrial estimates of the timescale of human evolution in the context of the contemporary palaeontological and archaeological evidence for key stages in human prehistory. We contend that some of the most widely-cited mitochondrial rate estimates have several significant shortcomings, including a reliance on a human-chimpanzee calibration, and highlight the pressing need for revised rate estimates. PMID- 19682766 TI - Influence of temporally variable groundwater flow conditions on point measurements and contaminant mass flux estimations. AB - Monitoring of contaminant concentrations, e.g., for the estimation of mass discharge or contaminant degradation rates, often is based on point measurements at observation wells. In addition to the problem, that point measurements may not be spatially representative, a further complication may arise due to the temporal dynamics of groundwater flow, which may cause a concentration measurement to be not temporally representative. This paper presents results from a numerical modeling study focusing on temporal variations of the groundwater flow direction. "Measurements" are obtained from point information representing observation wells installed along control planes using different well frequencies and configurations. Results of the scenario simulations show that temporally variable flow conditions can lead to significant temporal fluctuations of the concentration and thus are a substantial source of uncertainty for point measurements. Temporal variation of point concentration measurements may be as high as the average concentration determined, especially near the plume fringe, even when assuming a homogeneous distribution of the hydraulic conductivity. If a heterogeneous hydraulic conductivity field is present, the concentration variability due to a fluctuating groundwater flow direction varies significantly within the control plane and between the different realizations. Determination of contaminant mass fluxes is also influenced by the temporal variability of the concentration measurement, especially for large spacings of the observation wells. Passive dosimeter sampling is found to be appropriate for evaluating the stationarity of contaminant plumes as well as for estimating average concentrations over time when the plume has fully developed. Representative sampling has to be performed over several periods of groundwater flow fluctuation. For the determination of mass fluxes at heterogeneous sites, however, local fluxes, which may vary considerably along a control plane, have to be accounted for. Here, dosimeter sampling in combination with time integrated local water flux measurements can improve mass flux estimates under dynamic flow conditions. PMID- 19682767 TI - Effect of selenium on characteristics of rape chloroplasts modified by cadmium. AB - Selenium appears to be an important protective agent that decreases cadmium induced toxic effects in animals and plants. The aim of these studies was to investigate the changes of properties of chloroplast membranes obtained from Cd treated rape seedlings caused by Se additions. Chloroplasts were isolated from leaves of 3-week-old rape plants cultured on Murashige-Skoog media supplied with 2 microM Na(2)SeO(4) and/or 400 microM CdCl(2) under in vitro conditions. The following physicochemical characteristics of chloroplasts were chosen as indicators of Se-effects: average size, zeta potential, ultrastructure, lipid and fatty acid composition and fluidity of envelope membrane. The results suggest that Se can partly counterbalance the destructive effects of Cd. This protective action led to an increase of chloroplast size reduced by Cd treatment and rebuilt, to some extent, the chloroplast ultrastructure. Lipid and fatty acid composition of chloroplast envelopes modified by Cd showed a decrease in digalactosyl-diacylglycerol content and an increase of content of monogalactosyl diacylglycerol and phospholipid fractions, as well as an increase of fatty acid saturation of all lipids studied. The change in fatty acid saturation correlated well with a decrease of membrane fluidity and with a diminishing of absolute values of zeta potential. The presence of selenium in cultured media caused a partial reversal of the detected changes, which was especially visible in properties related to the hydrophobic part of an envelope, i.e. fatty acid saturation and fluidity. PMID- 19682768 TI - Expression and functional characterization of two pathogenesis-related protein 10 genes from Zea mays. AB - A novel PR10 gene (ZmPR10.1) was isolated from maize and its expression and function were compared with the previous ZmPR10. ZmPR10.1 shares 89.8% and 85.7% identity to ZmPR10 at the nucleotide and amino acid sequence level, respectively. ZmPR10 and ZmPR10.1 were mainly expressed in root tissue with low expression in other tissues. ZmPR10.1 had significantly lower expression than ZmPR10 in all tissues examined. The expression of both ZmPR10 and ZmPR10.1 was induced by most abiotic stresses including SA, CuCl(2), H(2)O(2), coldness, darkness and wounding during the 16-h treatments, and biotic stresses such as Erwinia stewartii and Aspergillus flavus infection. However, ZmPR10.1 was induced only 2 HAT and down regulated thereafter, whereas ZmPR10 remained induced during the 16-h NAA treatment. Also, inoculation with Erwinia chrysanthemi caused about 2-fold induction in ZmPR10.1 expression 60 HAT but not significant changes for ZmPR10. Both ZmPR10.1 and ZmPR10 showed RNase activity in vitro with an optimal pH and temperature of 6.5 and 55 degrees C. Their RNase activities were significantly inhibited by low concentrations (1.0mM) of Cu(2+), Ag(+), Co(2+), SDS, EDTA or DTT. However, ZmPR10.1 possessed significantly higher (8-fold) specific RNase activity than ZmPR10. Also, ZmPR10.1 showed a stronger inhibition against bacterium Pseudomonas syringae pv. tomato DC3000 in vivo and fungus A. flavus in vitro than ZmPR10, indicating that ZmPR10.1 may also play an important role in host plant defense. PMID- 19682769 TI - Phase I/II randomized bilateral half-head comparison of topical bexarotene 1% gel for alopecia areata. AB - BACKGROUND: Alopecia areata, hair loss caused by perifollicular T-cell infiltrates, is refractory to therapy. Bexarotene, a retinoid X receptor is a selective retinoid, induces T-cell apoptosis. OBJECTIVE: We sought to determine the safety, including the dose-limiting toxicities with adverse events, and efficacy, ie, response rate, of bexarotene in alopecia areata. METHODS: We conducted a phase I/II randomized, half-head trial of 1% bexarotene gel applied twice daily for 6 months. RESULTS: In all, 42 patients (11 male and 31 female) with alopecia totalis (n = 3), alopecia universalis (n = 5), or alopecia areata (n = 34) applied 1% bexarotene gel for 24 weeks. Five of 42 (12%) had 50% or more partial hair regrowth on the treated side, and 6 of 42 (14%) on both sides including 3 complete responders. In all, 31 patients had mild irritation; 4 had grade-3 irritation. LIMITATIONS: This design cannot differentiate between drug induced and spontaneous regrowth. CONCLUSION: Topical bexarotene 1% application is well tolerated and possibly effective. A randomized placebo-controlled trial should be conducted. PMID- 19682771 TI - Analgesic, anticonvulsant and anti-inflammatory activities of some synthesized benzodiazipine, triazolopyrimidine and bis-imide derivatives. AB - A series of diazipine, pyrimidine, fused triazolopyrimidine and imide derivatives were newly synthesized using 4-phenyl-but-3-en-2-one 1 as a starting material and compounds 2 and 9 are intermediates. Initially the acute toxicity of the compounds was assayed via the determination of their LD(50). All the compounds were interestingly less toxic than the reference drug. The pharmacological screening showed that many of these obtained compounds have good analgesic, anticonvulsant and anti-inflammatory activities comparable to Valdecoxib, Carbamazepine and Predensilone as reference drugs. Regarding the protection against Carrageenan induced edema, five compounds were found more potent than Prednisolone. On the other hand, in searching for COX-2 inhibitor, the inhibition of plasma PGE2 for the compounds were determined and four compounds were found more potent than Prednisolone. The detailed synthesis, spectroscopic data, pharmacological screening and acute toxicity LD(50) for the synthesized compounds were reported. PMID- 19682772 TI - Learning pathways in contemporary primary care settings - student nurses's views. AB - Workforce planning to address nursing supply along with curriculum change affecting practice placements has acted as key drivers shaping learning resources for student nurses within Aberdeen Community Health Partnership. As part of their learning, students are afforded the opportunity to visit a range of contemporary primary care settings such as public health, specialist nurses and NHS 24 on what are known as 'pathway experiences'. This research adopted a qualitative approach using focus group interviews to discover students' learning as a result of this initiative. Two main themes emerged, these being 'Healthcare contexts' and 'Nursing know-how'. Students developed a greater awareness and understanding about the delivery of healthcare in primary care settings and they expanded their nursing knowledge and skills. While students valued the experience, they also thought pathways fragmented their time at the base placement and detracted from gaining the practical nursing skills they considered of prime importance in their transit towards qualifying and working within a clinical setting. Implications for the future development of the pathway initiative are discussed. PMID- 19682773 TI - Who enters nursing schools and why do they choose nursing? A comparison with female non-nursing students using longitudinal data. AB - Understanding who enters nursing schools and why they choose nursing is essential for the nursing profession to recruit and retain their successors. This study was conducted to examine the characteristics of nursing students and factors influencing their career decision, as compared with non-nursing students. We used public databases of the Korean Education & Employment Panel that followed 4000 high school students in their final year of school. Out of those students, 2456 students entered a college or university and 40 students (39 females and one male) entered a nursing school. Nursing students were compared with 1011 female non-nursing students. Nursing students had lower household income. Nursing students had better high school academic achievement overall and in science. A more studious attitude and lower absenteeism were found among nursing students. For nursing students, employability (55%) followed by aptitude (20%) were the most influential factors in choosing a nursing major, while conversely, aptitude (48%) followed by employability (23%) were most important to non-nursing students. About half of nursing students answered "herself/himself" as the most influential person in choosing nursing, whereas three quarters of non-nursing students did. Nursing students had a higher rate (75%) of satisfaction with their major than non-nursing students (60%). PMID- 19682770 TI - Local media influence on opting out from an exception from informed consent trial. AB - STUDY OBJECTIVE: News media are used for community education and notification in exception from informed consent clinical trials, yet their effectiveness as an added safeguard in such research remains unknown. We assessed the number of callers requesting opt-out bracelets after each local media report and described the errors and content within each media report. METHODS: We undertook a descriptive analysis of local media trial coverage (newspaper, television, radio, and Web log) and opt-out requests during a 41-month period at a single site participating in an exception from informed consent out-of-hospital trial. Two nontrial investigators independently assessed 41 content-based media variables (including background, trial information, graphics, errors, publication information, and assessment) with a standardized, semiqualitative data collection tool. Major errors were considered serious misrepresentation of the trial purpose or protocol, whereas minor errors included misinformation unlikely to mislead the lay reader about the trial. We plotted the temporal relationship between opt-out bracelet requests and media reports. Descriptive information about the news sources and the trial coverage are presented. RESULTS: We collected 39 trial related media reports (33 newspaper, 1 television, 1 radio, and 4 blogs). There were 13 errors in 9 (23%) publications, 7 of which were major and 6 minor. Of 384 requests for 710 bracelets, 310 requests (80%) occurred within 4 days after trial media coverage. Graphic timeline representation of the data suggested a close association between media reports about the trial and requests for opt-out bracelets. CONCLUSION: According to results from a single site, local media coverage for an exception from informed consent clinical trial had a substantial portion of errors and appeared closely associated with opt-out requests. PMID- 19682774 TI - Faculty mentors' and students' perceptions of students' research self-efficacy. AB - Mentoring in nursing is an important process for socializing nurse researchers, developing a body of professional knowledge, and influencing career choices of students. Self-efficacy (Bandura, 1997) is concerned with one's perceived ability to perform tasks within a specific domain. The purpose of this study was to compare undergraduate and graduate student's perceptions of their abilities to pursue research (research self-efficacy) with their mentors' perceptions. A cross sectional design was used to study mentors in any academic discipline who received external funding and worked with an undergraduate or graduate student on the research study. Recruitment and data collection were completed using the Internet and included 21 faculty mentors and student dyads. The Clinical Research Appraisal Inventory was used to measure research self-efficacy. Differences between the faculty mentor's perception of the student's confidence in research and students' perception were significant at p=<0.001. Misjudgment of efficacy appraisals can result in opportunities forsaken and careers not pursued. Assisting mentors to guide students' skill perfection may increase students' choice of research careers, promote the effectiveness of mentorship, aid in the development of a body of professional knowledge and benefit careers of both mentors and students. PMID- 19682775 TI - Integrating problem-based learning in a nursing informatics curriculum. AB - In recent years employers in health care organizations have been recognizing the need for nurses to enter the workforce with a set of informatics competencies. Numerous nursing informatics programs have been established worldwide. The challenge becomes to explore innovative tools that will equip nurses with the appropriate skills to utilize information technology to improve health care quality and patient safety and redesign health care services. This paper presents the introduction of problem-based learning (PBL) modules into an existing nursing informatics curriculum, the Clinical Informatics and Patient Centered Technologies Master program at the School of Nursing, University of Washington. Additionally, we discuss recommendations and challenges associated with the integration of PBL in nursing informatics graduate education including the need for facilitators, flexible technology platforms, promotion and documentation of group work, faculty training and supervision by a program committee. PMID- 19682777 TI - Isolated posterior orbital floor fractures, diplopia and oculocardiac reflexes: a 10-year review. AB - Isolated fractures of the posterior orbital floor are rarely encountered in facial trauma, but warrant appropriate management to afford optimum recovery from debilitating diplopia and symptoms of activation of the oculocardiac reflex. We reviewed all records of patients with isolated fractures of the orbital floor between 1997 and 2007. Seven of 58 fractures operated on during this time involved the posterior orbit alone (all male, age range: 10-23). These seven patients presented with serious diplopia in the primary gaze, intense orbital pain, and a range of signs and symptoms of activation of the oculocardiac reflex. Early operations were done in all cases (usually within 24h), which immediately relieved pain and the effects of the oculocardiac reflex. Postoperatively diplopia resolved substantially in upgaze but was worse in downgaze, and took between 2 and 6 months to resolve to normal functional ranges, which were confirmed by a Hess chart. This review suggests that this fracture pattern is not restricted to children as is often reported, and operation should be within 24 48h of injury to optimise outcome. Diplopia will improve postoperatively, but will persist over months, and patients should be well informed of this. Interestingly none of our patients' injuries were the result of assault, even though this accounted for nearly 75% of all facial fractures treated in our department. Five of the seven patients were adults, and the mechanism of fracture pointed to being crushed by injuries of low velocity and high force. PMID- 19682776 TI - Dutch national survey to test the STRONGkids nutritional risk screening tool in hospitalized children. AB - BACKGROUND & AIMS: Children admitted to the hospital are at risk of developing malnutrition. The aim of the present study was to investigate the feasibility and value of a new nutritional risk screening tool, called STRONG(kids), in a nationwide study. METHODS: A Prospective observational multi-centre study was performed in 44 Dutch hospitals (7 academic and 37 general), over three consecutive days during the month of November 2007. The STRONG(kids) screening tool consisted of 4 items: (1) subjective clinical assessment, (2) high risk disease, (3) nutritional intake, (4) weight loss. Measurements of weight and length were performed. SD-scores <-2 for weight-for-height and height-for-age were considered to indicate acute and chronic malnutrition respectively. RESULTS: A total of 424 children were included. Median age was 3.5 years and median hospital stay was 2 days. Sixty-two percent of the children were classified "at risk" of developing malnutrition by the STRONG(kids) tool. Children at risk had significantly lower SD-scores for weight-for-height, a higher prevalence of acute malnutrition and a longer hospital stay compared to children with no nutritional risk. CONCLUSIONS: The nutritional risk screening tool STRONG(kids) was successfully applied to 98% of the children. Using this tool, a significant relationship was found between having a "high risk" score, a negative SD-score in weight-for-height and a prolonged hospital stay. PMID- 19682778 TI - Belowground effects of enhanced tropospheric ozone and drought in a beech/spruce forest (Fagus sylvatica L./Picea abies [L.] Karst). AB - The effects of experimentally elevated O(3) on soil respiration rates, standing fine-root biomass, fine-root production and delta(13)C signature of newly produced fine roots were investigated in an adult European beech/Norway spruce forest in Germany during two subsequent years with contrasting rainfall patterns. During humid 2002, soil respiration rate was enhanced under elevated O(3) under beech and spruce, and was related to O(3)-stimulated fine-root production only in beech. During dry 2003, the stimulating effect of O(3) on soil respiration rate vanished under spruce, which was correlated with decreased fine-root production in spruce under drought, irrespective of the O(3) regime. delta(13)C signature of newly formed fine-roots was consistent with the differing g(s) of beech and spruce, and indicated stomatal limitation by O(3) in beech and by drought in spruce. Our study showed that drought can override the stimulating O(3) effects on fine-root dynamics and soil respiration in mature beech and spruce forests. PMID- 19682779 TI - Urinary arsenic species and CKD in a Taiwanese population: a case-control study. AB - BACKGROUND: Inorganic arsenic has been linked to decreased kidney function through oxidative damage. Arsenic methylation is believed to be a pathway for arsenic metabolism. Lycopene is an antioxidant that reduces oxidative stress; however, the association between urinary arsenic species, plasma lycopene level, and chronic kidney disease (CKD) has seldom been evaluated. STUDY DESIGN: Case control study. SETTING & PARTICIPANTS: 125 patients with CKD and 229 controls were recruited from a hospital-based pool. PREDICTOR: Urinary arsenic species and plasma lycopene level. OUTCOMES & MEASUREMENTS: CKD was defined as estimated glomerular filtration rate (eGFR) less than 60 mL/min/1.73 m(2), calculated by using the Modification of Diet in Renal Disease Study equation. Plasma lycopene was measured by means of high-performance liquid chromatography. Urinary arsenic species, including arsenite, arsenate, monomethylarsonic acid, and dimethylarsinic acid, were determined by means of high-performance liquid chromatography and hydride generator-atomic absorption spectrometry. RESULTS: Lycopene level was associated positively with eGFR, and participants with a high serum lycopene level had a significant, inverse association with CKD (odds ratio, 0.41; 95% confidence interval, 0.21 to 0.81). Total arsenic level was associated significantly with CKD in a dose-response relationship, especially in participants with a total arsenic level greater than 20.74 compared with 11.78 microg/g creatinine or less (odds ratio, 4.34; 95% confidence interval, 1.94 to 9.69). Furthermore, participants with a high urinary total arsenic level or participants with a low percentage of dimethylarsinic acid had a positive association with CKD when their plasma lycopene level was low. LIMITATIONS: Because of the single spot evaluation of plasma antioxidants and urinary arsenic species and the small sample size, statistical significance should be interpreted with caution. CONCLUSIONS: This study shows that high urinary total arsenic or low plasma lycopene level is associated positively with CKD. Results suggest that the capacity for arsenic methylation may be associated with CKD in individuals who ingest low arsenic levels in drinking water and also have a low plasma lycopene level. PMID- 19682780 TI - Induction antibody therapy in kidney transplantation. AB - Antilymphocyte antibodies have been used for the prevention or treatment of acute rejection in kidney transplant recipients since the 1960s. Both monoclonal and polyclonal agents now are available and generally are classified as either lymphocyte-depleting or nondepleting agents. Use of such antibodies for induction therapy in the immediate postoperative period has varied over the years. Currently, induction antibodies are administered to more than 70% of kidney transplant recipients in the United States. However, the choice of specific agents and the patients for whom they are used vary substantially between and within transplant centers. Many centers use antibody induction therapy only in patients perceived to be at high risk of acute rejection or delayed graft function. Recently, induction antibody therapy also has become the standard of practice in protocols designed to facilitate minimization of such maintenance immunosuppressive drugs as corticosteroids or calcineurin inhibitors. The benefits of induction therapy, including a decreased incidence and delayed onset of acute rejection, must be balanced against the considerable cost and side effects of the individual agents, including risk of infection. Some, but not all, antibodies are associated with increased risk of posttransplantation lymphoproliferative disease and other malignancies. PMID- 19682782 TI - Copycat effects after media reports on suicide: a population-based ecologic study. AB - This study aimed to investigate whether the risk of increased suicide occurrence after reports on suicide is associated with the social characteristics of the reported suicides and whether this varies with similarity between the reported suicides and suicides in the population. We collected reports on all 179 individual suicides named in the 13 largest Austrian nationwide newspapers from 1996 to 2006. Information on social status and sociodemographic characteristics of the reported suicides, on certainty of labelling the death as a suicide, and on the suicide methods applied were extracted from the articles. We conducted logistic regression analyses, with the increase of post-report suicides within 28 days after the reports as dependent variable. In model 1, the increase of suicides that matched the reported individual suicide with regard to age group, sex and suicide method was used as outcome variable. In model 2, the increase of suicides that were different from the reported suicide with regard to these characteristics was the outcome. In model 3, the post-report increase of total suicides was the dependent variable. Celebrity status of the reported suicide, age of the reported suicide between 30 and 64 years, and definitive labelling as a suicide were associated with an increased risk of a post-report increase of similar suicides; criminality (i.e. the individual was reported as suspected or convicted of crime) of the reported suicide was associated with a lower risk of a post-report increase. In dissimilar suicides, none of the variables was associated with a post-report increase of suicides. Celebrity status of the reported suicide was the only predictor of a post-report increase of total suicides. The findings support the hypothesis that social variables of reported suicides impact the risk of post-report copycat behaviour. Evidence of copycat effects seemed to be strongest in suicides that were similar to the respective model with regard to age group, sex, and suicide method. PMID- 19682781 TI - The influence of emotion-focused rumination and distraction on depressive symptoms in non-clinical youth: a meta-analytic review. AB - This review examined evidence for some core predictions of the response styles theory (RST) concerning the relation between response styles and symptoms of depression and gender differences in the use of response styles in non-clinical children and adolescents. In summarizing the literature, effect sizes (pooled correlation coefficients) were calculated for cross-sectional and longitudinal studies. Stability of the obtained effect sizes was evaluated by means of a fail safe N analysis. Results indicated that stable and significant effect sizes were found for rumination being associated with concurrent and future levels of depression. When controlling for baseline levels of depression, effect sizes for rumination and distraction were not stable, indicating that these findings should be interpreted with considerable caution. Finally, significant and stable effect sizes for gender differences in response styles were found only for rumination among adolescents. Taken together, the findings partly support the predictions of the response styles theory examined in this meta-analysis and may implicate that rumination is a cognitive vulnerability factor for depressive symptoms among adolescents. PMID- 19682784 TI - [Lipofilling (for reconstructive surgery after breast cancer): a new technique for fat recuperation]. PMID- 19682783 TI - [Muscle-sparing latissimus dorsi flap. Vascular anatomy and indications in breast reconstruction]. AB - BACKGROUND: The muscle-sparing latissimus dorsi flap pedicled on descending branch presents distinct advantages in breast reconstruction, specially when there is a transversely oriented skin paddle, including reduced donor site morbidity, sparing muscle function and greater freedom of orientation of the skin paddle. This study reports the anatomical basis, surgical technique, advantages and complications of this technique. Four clinical cases illustrate surgical indications in breast reconstructive surgery. METHODS: An anatomical cadaveric study underwent to University of Texas Southwestern Medical Center, Dallas. The goal was performed to determine the location of the bifurcation of the thoracodorsal artery and the course of its descending branch compare to the anterior side of latissimus dorsi muscle. Four clinical cases illustrated indications of muscle-sparing latissimus dorsi flap pedicled on descending branch in breast reconstruction. These cases showed advantages and complications of the technique, and impact on donor site. RESULTS: Fifteen descending branch muscle sparing latissimus dorsi flaps were harvested. All flaps had a bifurcation of the thoracodorsal artery. The average was located at 5,1cm from posterior axillary side (from 2,1 to 7,5 cm) and average of 2,2 cm from the anterior side of latissimus dorsi muscle (from 1,3 to 3,1cm). To 5, 10 and 15 cm from posterior axillary side, the descending branch was located at respectively an average of 2,0 cm (from 1,4 to 2,5), 2,4 cm (from 1,3 to 3,3), and 2,9 cm (from 2,0 to 3,8) behind the anterior side of latissimus dorsi muscle. The average length of descending branch was measured at 15,2 cm (from 13,2 to 19,0). None clinical cases paddle suffering was observed. Donor site morbidity was less than classical or extended adipomuscular technique. Latissimus dorsi muscle function is spared. CONCLUSIONS: The muscle-sparing latissimus dorsi flap, pedicled on descending branch, is versatile and reproducible. It results in minimal functional deficit of the donor site, absence of seroma, large freedom of orientation of the skin paddle, low rate of flap complications, and a cosmetically acceptable scar. There are a lot of indications in breast reconstruction. PMID- 19682786 TI - [Analytical study of the silhouette: the segmental syndromes and therapeutic options]. AB - Many studies showed we have a visceral adipose tissue and a subcutaneous adipose tissue. Roles and risks of these fats are different. The purpose of this work is to demonstrate that there are at least two types of subcutaneous fat; one is "metabolic" and diet sensitive and another genetic which is diet resistant. The distribution of subcutaneous fats is also determined by genetic factors, and its study led to the description of segmental syndromes whose distribution and evolution were studied in women in our environment. It seems that no fundamental studies have been made on these two types of fat. We think there is also a genetic programming of our weight which varies with age. This programming may be modulated by the environment (physical activity, diet) but always on a temporary basis. In recent years, the surgical treatment of obesity by gastric rings or by gastric bypass was right to cite, while liposuction is still considered as an accessory process, reserved for purely cosmetic indications. In light of the above, we review the preventive and therapeutic indications of this technique. PMID- 19682785 TI - [Benefit of complementary lipofilling in aesthetic breast augmentation with implant]. AB - Since 2 years, the author uses an aesthetic mammary augmentation technique with a combination in the same surgical procedure of: placement of a breast implant in a retrofascial plane; lipofilling under the skin of the cleavage to cover the upper part of the implant. Forty-seven women have been operated with this procedure since 2006, with immediate satisfactory results in all the cases. We consider this procedure as an interesting alternative to retromuscular placement in thin patients with small breast. PMID- 19682788 TI - Real-time contrast-enhanced ultrasound for the assessment of perfusion dynamics in skeletal muscle. AB - We developed a real-time low-MI contrast-enhanced ultrasound method (CEUS), compared it with venous occlusion plethysmography (VOP) and evaluated its robustness in the quantification of skeletal muscle perfusion during exercise. Contrast pulse sequencing (7 MHz) during continuous intravenous infusion of SonoVue (4.8 mL/300 s) was used repeatedly in eight healthy volunteers to monitor changes of the muscle perfusion before, during and after isometric exercises (10 to 50% of individual maximum strength for 20 to 30 s) of the gastrocnemius muscle in real time. CEUS was correlated with VOP at different time points, and the exactness of several CEUS parameters obtained from ultrasound-signal-intensity time curves was evaluated. Real-time CEUS depicted a large variability of the skeletal muscle blood volume at rest (mean, 3.48; range, 0.60 to 9.92 [approximately mL]), with a significant reproducibility (r=0.72, p<0.05) and correlation with VOP (r=0.59, p<0.001). Mean blood volume during exercise was 1.58(approximately mL), increased to a mean maximum after exercise of 8.88 (approximately mL), the mean change of the local blood volume during and directly after the exercise was -0.10 and +1.57(approximately mL/s). The average CEUS signal during exercise decreased (mean area under the curve, -50.4 [approximately mL.s]) and subsequently increased post exercise (mean 118.6 [approximately mL.s]). CEUS parameters could be calculated with mean relative errors between 6 and 36%. Continuous assessment of local muscle microcirculation during exercise is possible with real-time CEUS with an acceptable robustness. Its application may be of particular interest in a better understanding of the role of perfusion during muscle training, and the monitoring of pathological vascular response, such as in diabetic microvessel diseases. PMID- 19682787 TI - Subcutaneous graft of D1 mouse mesenchymal stem cells leads to the formation of a bone-like structure. AB - Mesenchymal stem cells (MSC) are capable of both self-renewal and multi-lineage differentiation into mesoderm-type cells such as osteoblasts, chondrocytes, adipocytes and myocytes. Together the multipotent nature of MSCs and the facility to expand them in vitro make these cells ideal resources for regenerative medicine, particularly for bone reconstruction, and therefore research efforts focused on defining efficient protocols for directing their differentiation into the requisite lineage. Despite much progress in identifying mechanisms and factors that direct and control in vitro osteogenic differentiation of MSCs, a rapid and simple model to evaluate in vivo tissue formation is still lacking. Here, we describe the unique capacity of the murine bone marrow-derived D1 MSC cell line, which differentiates in vitro into at least three cell lineages, to form in vivo a structure resembling bone. This bone-like structure was obtained after subcutaneous grafting of D1 cells into immunocompetent mice without the need of neither an osteogenic factor nor scaffold material. These data allow us to propose this cell model as a tool for exploring in vivo the mechanisms and/or factors that govern and potentially regulate osteogenesis. PMID- 19682789 TI - A generalized speckle tracking algorithm for ultrasonic strain imaging using dynamic programming. AB - This study developed an improved motion estimation algorithm for ultrasonic strain imaging that employs a dynamic programming technique. In this article, we model the motion estimation task as an optimization problem. Since tissue motion under external mechanical stimuli often should be reasonably continuous, a set of cost functions combining correlation and various levels of motion continuity constraint were used to regularize the motion estimation. To solve the optimization problem with a reasonable computational load, a dynamic programming technique that does not require iterations was used to obtain displacement vectors in integer precision. Then, a subsample estimation algorithm was used to calculate local displacements in fractional precision. Two implementation schemes were investigated with in vivo ultrasound echo data sets. We found that the proposed algorithm provides more accurate displacement estimates than our previous algorithm for in vivo clinical data. In particular, the new algorithm is capable of tracking motion in more complex anatomy and increases strain image consistency in a sequence of images. Preliminary results also suggest that a significantly longer sequence of high contrast strain images could be obtained with the new algorithm compared with the previous algorithm. The new algorithm can also tolerate larger motion discontinuities (e.g., cavity in an anthropomorphic uterine phantom). PMID- 19682791 TI - Biodegradation of the low concentration of polycyclic aromatic hydrocarbons in soil by microbial consortium during incubation. AB - The biodegradation of polycyclic aromatic hydrocarbons (PAHs) (8.15 mg PAHs kg( 1) soil) in aged contaminated soil by isolated microbial consortium (five fungi and three bacteria) during the incubation of 64d is reported. The applied treatments were: (1) biodegradation by adding microbial consortium in sterile soils (BM); (2) biodegradation by adding microbial consortium in non-sterile soils (BMN); and (3) biodegradation by in situ "natural" microbes in non-sterile soils (BNN). The fungi in BM and BMN soils grew rapidly 0-4d during the incubation and then reached a relative equilibrium. In contrast the fungi in BNN soil remained at a constant level for the entire time. Comparison with the fungi, the bacteria in BNN soils grew rapidly during the incubation 0-2d and then reached a relative equilibrium, and those in BM and BMN soils grew slowly during the incubation of 64 d. After 64 d of incubation, the PAH biodegradations were 35%, 40.7% and 41.3% in BNN, BMN and BM, respectively. The significant release of sequestrated PAHs in aged contaminated soil was observed in this experiment, especially in the BM soil. Therefore, although bioaugmentation of introduced microbial consortium increased significantly the biodegradation of PAHs in aged contaminated soil with low PAH concentration, the creation of optimum of the environmental situation might be the best way to use bioremediation successfully in the field. PMID- 19682790 TI - Prostate-specific antigen velocity for early detection of prostate cancer: result from a large, representative, population-based cohort. AB - BACKGROUND: It has been suggested that changes in prostate-specific antigen (PSA) over time (ie, PSA velocity [PSAV]) aid prostate cancer detection. Some guidelines do incorporate PSAV cut points as an indication for biopsy. OBJECTIVE: To evaluate whether PSAV enhances prediction of biopsy outcome in a large, representative, population-based cohort. DESIGN, SETTING, AND PARTICIPANTS: There were 2742 screening-arm participants with PSA <3 ng/ml at initial screening in the European Randomized Study of Screening for Prostate Cancer in Rotterdam, Netherlands, or Goteborg, Sweden, and who were subsequently biopsied during rounds 2-6 due to elevated PSA. MEASUREMENTS: Total, free, and intact PSA and human kallikrein 2 were measured for 1-6 screening rounds at intervals of 2 or 4 yr. We created logistic regression models to predict prostate cancer based on age and PSA, with or without free-to-total PSA ratio (%fPSA). PSAV was added to each model and any enhancement in predictive accuracy assessed by area under the curve (AUC). RESULTS AND LIMITATIONS: PSAV led to small enhancements in predictive accuracy (AUC of 0.569 vs 0.531; 0.626 vs 0.609 if %fPSA was included), although not for high-grade disease. The enhancement depended on modeling a nonlinear relationship between PSAV and cancer. There was no benefit if we excluded men with higher velocities, which were associated with lower risk. These results apply to men in a screening program with elevated PSA; men with prior negative biopsy were not evaluated in this study. CONCLUSIONS: In men with PSA of about >=3 ng/ml, we found little justification for formal calculation of PSAV or for use of PSAV cut points to determine biopsy. Informal assessment of PSAV will likely aid clinical judgment, such as a sudden rise in PSA suggesting prostatitis, which could be further evaluated before biopsy. PMID- 19682792 TI - Pyridine degradation in the microbial fuel cells. AB - The objective of this study was to investigate the feasibility of pyridine biodegradation in the microbial fuel cell (MFC), from which electricity was generated. Experiments were initially conducted in a graphite-packed MFC (G-MFC) using a pyridine concentration of 500 mg/L combined with different glucose concentrations. Pyridine of 500 mg/L only used as the G-MFC fuel resulted in a maximal voltage of 116 mV and a maximal power density of 1.7 W/m(3). The maximal voltage reached within 12 h when pyridine was totally depleted. The glucose supplement with concentrations of 500, 250, and 100 mg/L resulted in the maximum voltages of 623, 538, and 349 mV, respectively, correspondingly the maximal volumetric power densities were 48.5, 36.2, and 15.2 W/m(3). Pyridine biodegradation rates reached 95% within 24h using the G-MFC. Interestingly, after 90 d of acclimation, the biodegradation rates of pyridine in the G-MFC using pyridine only as the fuel were higher than those using the glucose-pyridine mixtures. Further experiments were conducted using a graphite fiber brush MFC (B MFC). Compared to the G-MFC, the B-MFC enhanced the electrical charges by 89, 186, and 586% for the mixtures with ratios of glucose-to-pyridine of 1:1, 1:2, and 1:5, respectively. GC/MS analyses of the anode solution indicated that the metabolism of pyridine in the MFC was initiated by ring reduction and NH3-N production. The results suggest that pyridine may be used as the MFC fuel in practical applications of wastewater treatment. PMID- 19682793 TI - Endogenous opioids may buffer effects of anger arousal on sensitivity to subsequent pain. AB - Evidence suggests that anger and pain are related, yet it is not clear by what mechanisms anger may influence pain. We have proposed that effects of anger states and traits on pain sensitivity are partly opioid mediated. In this study, we test the extent to which analgesic effects of acute anger arousal on subsequent pain sensitivity are opioid mediated by subjecting healthy participants to anger-induction and pain either under opioid blockade (oral naltrexone) or placebo. Participants were 160 healthy individuals. A double blind, placebo-controlled, between-subjects opioid blockade design is used, with participants assigned randomly to one of two drug conditions (placebo or naltrexone), and to one of two Task Orders (anger-induction followed by pain or vice versa). Results of ANOVAs show significant Drug Condition x Task Order interactions for sensory pain ratings (MPQ-Sensory) and angry and nervous affect during pain-induction, such that participants who underwent anger-induction prior to pain while under opioid blockade (naltrexone) reported more pain, and anger and nervousness than those who underwent the tasks in the same order, but did so on placebo. Results suggest that for people with intact opioid systems, acute anger arousal may trigger endogenous opioid release that reduces subsequent responsiveness to pain. Conversely, impaired endogenous opioid function, such as that found among some chronic pain patients, may leave certain people without optimal buffering from the otherwise hyperalgesic affects of anger arousal, and so may lead to greater pain and suffering following upsetting or angry events. PMID- 19682794 TI - Latency to facial expression change following noxious stimulation in infants is dependent on postmenstrual age. AB - Change in facial expression over a fixed time after a noxious stimulus is the key measure used to calculate pain scores in preterm and newborn infants. We hypothesised that the latency of facial motor responses would be longer in the youngest premature infants and that behavioural scoring methods of pain may need to take this into account. One hundred and seventy-two clinically required heel lances were performed in 95 infants from 25 to 44 weeks postmenstrual age (PMA). Sixty-four percentage of the heel lances evoked a change in facial expression. Change in facial expression was observed in infants across the whole age range from 25 weeks PMA and the latency to the facial expression response ranged from 1 to 17s. Latency to facial expression change was dependent on the infants' PMA at the time of the heel lance. Infants below 32 weeks PMA had a significantly longer latency to change in facial expression than older infants (54% increase in infants below 32 weeks; p < 0.001). Sleep state and presence of brain damage (IVH grades 1-4) did not significantly increase the latency (p > 0.05 for each variable). Intravenous morphine at the time of the heel lance significantly increased the latency to facial expression response (p < 0.001) but the analysis shows that latency is highly dependent on PMA independent of morphine administration. These findings highlight developmental changes underlying infant behaviour that are critically important if pain scores are to be correctly interpreted. PMID- 19682796 TI - Identification of cryptic species of Culicoides (Diptera: Ceratopogonidae) in the subgenus Culicoides and development of species-specific PCR assays based on barcode regions. AB - Culicoides biting midges (Diptera: Ceratopogonidae) are vectors of important diseases affecting wild and domestic animals. During the last decade they have played a major role in the epidemiology of the largest bluetongue epizootic ever recorded in Europe, the disease is transmitted between hosts almost exclusively by bites of Culicoides midges and affects both domestic and wild ruminants however severe disease usually occurs in certain breeds of sheep and some species of deer. An accurate vector identification is of major importance in arthropod borne diseases surveillance, as great differences in vectorial capacity are found even between close species. Unfortunately, specialized taxonomic knowledge of Culicoides identification is rarely available in routine surveillance, mainly based on wing morphology. Recently, some European species of Culicoides belonging to the subgenus Avaritia Fox, 1955 and Culicoides Latreille, 1809 have been described as new bluetongue virus vectors. In the present study, by using a fragment of the barcode region (COI gene) we report the presence of up to 11 species within the subgenus Culicoides in Catalonia (NE Spain), a region recently affected by a bluetongue epizootic. The molecular analysis revealed new non described cryptic species which were grouped in three complexes of morphologically similar species, two in the Pulicaris complex resembling Culicoides pulicaris, two in the Fagineus complex resembling Culicoides fagineus and three in the Newsteadi complex resembling Culicoides newsteadi. The phylogenetic relationships among them showed that cryptic species detected in both Pulicaris and Fagineus complexes were closely related, whereas those in the Newsteadi complex were more distant. Accurate analysis of all species using morphological and molecular approaches resulted in the detection of diagnostic metric traits for cryptic species and the design of several new species-specific single and multiplex PCR assays to identify unambiguously all the species, most of them still lacking a specific molecular diagnosis. PMID- 19682795 TI - Association between nematode larvae and "low worm egg count diarrhoea" in sheep in Western Australia. AB - Nine flocks of sheep with a high prevalence (>30%) of diarrhoea and severe breech faecal soiling were investigated over a three-year period to examine the causes of diarrhoea in sheep with low mean faecal worm egg counts (WEC). All nine flocks were located in the southwest of Western Australia in areas with a winter rainfall pattern (Mediterranean climate). There was no difference (p=0.304) in WEC of diarrhoeic sheep (loose faeces and severe breech faecal soiling) and "normal sheep" (pelleted faeces and mild or no breech faecal soiling). Teladorsagia (Ostertagia) circumcincta and Trichostrongylus spp. were the nematodes most commonly identified by total worm counts and differentiation of larvae recovered from faeces and pasture. Larval stages of strongyle worms accounted for the largest proportion of total worm counts in both diarrhoeic and normal sheep. Adult worm burdens were small in most sheep. Diarrhoeic sheep had higher numbers of fourth stage larvae than normal sheep (p=0.046). There was no histopathological evidence of bacterial or viral causes of diarrhoea in any of the flocks or bacteriological evidence of bacterial infections associated with diarrhoea. Two flocks had marginal selenium glutathione peroxidase (selenium) levels. One flock was diagnosed with helminthosis based on rising WEC and high total worm counts. Larval hypersensitivity diarrhoea, nutritional factors or a combination of these two factors were the most likely causes of diarrhoea in the other eight flocks based on exclusion of other known causes of diarrhoea. Treatment with moxidectin and an ivermectin controlled-release capsule did not change faecal moisture content of treated sheep compared to untreated sheep three to five weeks after treatment. The findings suggest that the immune response to strongyle larvae may explain some cases of low WEC diarrhoea observed during winter-spring in immunocompetent mature sheep grazing in Mediterranean environments. PMID- 19682798 TI - Screening for the presence of nematophagous fungi collected from Irish sheep pastures. AB - With worldwide development of anthelmintic resistance, alternative approaches to the chemotherapeutic dominant approach for the control of parasitic nematodes in sheep are urgently required. As natural enemies of nematodes, nematophagous fungi offer the exciting possibility of an alternative to the dominant anthelmintic approach for parasite control in ruminants. Permanent sheep pasture harbor a promising array of nematophagous fungi and merits further investigation. One hundred and fifty samples of soil, old and fresh faeces were collected from 10 Irish sheep pastures. The three methods employed for the isolation of nematophagous fungi include the Baermann technique, flotation method and the sprinkling-baiting technique. Twenty-nine nematophagous fungi were observed of which 12 were predacious and 17 were endoparasitic. The most prevalent fungi were Cystopage lateralis, Stylopage hadra, Drechmeria coniospora and Meristacrum asterosperum. Permanent sheep pasture is a good source of nematophagous fungi and hence may harbor potential biological control agents. Monacrosporium cionopaga, Duddingtonia flagrans, D. coniospora and Hirsutella rhoissilensis were detected in fresh faecal samples indicating they may have survived the gastrointestinal tract and therefore a viable option as a biological control agent. PMID- 19682797 TI - Non-curative, but prophylactic effects of paromomycin in Histomonas meleagridis infected turkeys and its effect on performance in non-infected turkeys. AB - Histomonosis (blackhead or infectious enterohepatitis) is a disease of gallinaceous birds, especially of turkeys, and is caused by the protozoan Histomonas meleagridis. Since the ban of all chemoprophylactic and chemotherapeutic products against this disease in the European Union, this parasite causes a considerable amount of economical problems in the poultry industry. Research which could ultimately lead to the discovery of new drugs against this disease is thus highly necessary. Hence, in this study, the efficacy of paromomycin against histomonosis in turkeys was investigated. First, the prophylactic and therapeutic efficacy of this drug against H. meleagridis and its effect on the weight gain of turkeys was determined. Adding paromomycin to the feed (400 ppm as well as 200 ppm paromomycin) or to the drinking water (420 mg paromomycin per liter water, added prior to or on the day of challenge) significantly lowered the mortality rate and the caecal and liver lesion scores after an intracloacal infection compared to infected untreated birds. However, when paromomycin was administered to turkeys in the drinking water after the challenge, no significant differences in mortality or in lesion scores could be observed compared to the infected untreated control group. This demonstrates that paromomycin exerts a purely preventive action against histomonosis in turkeys. Additionally, the weight gain of the treated birds was positively influenced by the use of the drug, as the average weight gain of all treated groups (except for the group treated at the day of first mortality) was significantly higher than that of the untreated control group. Finally, the target site of paromomycin was detected in the SS rRNA gene of H. meleagridis. Consequently, the susceptibility to paromomycin can be correlated to the presence of the binding site of the drug at the 3' end of the small subunit rRNA gene of the parasite. In conclusion, paromomycin can be used as a new prophylactic measure in the control of histomonosis in turkeys. PMID- 19682799 TI - Spatial distribution of Dermacentor reticulatus tick in Slovakia in the beginning of the 21st century. AB - A new field survey monitoring the spatial distribution of Dermacentor (D.) reticulatus (Fabricius, 1794) tick in Slovakia was carried out in 2005-2008 in order to record changes in its distribution when compared to former studies. Last surveys on the geographical distribution were conducted in 1950s and 1970s and the presence of D. reticulatus was determined along the rivers in the south-east (Latorica) as well as in the south-west (Morava, Dunaj) Slovakia. In the present survey new areas with D. reticulatus occurrence were detected, providing evidence that this tick species has extended its range in the surroundings of its former habitats but also by at least 200 km further North and by 300 m of elevation into higher altitudes. D. reticulatus is known to transmit Babesia spp. causing babesiosis in cattle and dogs. Expansion of D. reticulatus range is therefore likely to bring a spread of babesiosis, which can be severe or fatal especially for dogs. PMID- 19682800 TI - Intestinal and haematic parasitism in the birds of the Almunecar (Granada, Spain) ornithological garden. AB - Birds from the Almunecar ornithological garden (Granada, Spain) were surveyed from June 2006 to May 2007 to establish programmes to prevent, control, and treat intestinal and haematic parasites. A total of 984 faecal samples and 41 samples of blood were collected from Psittacidae, Cacatuidae, Phasianidae, and Anatidae. One or more intestinal parasites were identified in 51.6% of the samples. Blood parasites were found in 26.8% of the birds examined. The most frequent pathogenic endoparasites were coccidians, such as Cyclospora sp. (4.5%), Eimeria sp. (4.1%) and Isospora sp. (2%) and helminths such as Capillaria sp. (10. 1%), Ascaridia sp. (4.9%) and Heterakis gallinarum (4.9%). All the parasites varied with season but the most were found year round. Multiple parasitic infections by intestinal parasites were common, with 196 of 984 faecal samples having 2-5 intestinal parasites. The most frequent cases of multiple parasitism were Blastocystis plus Entamoeba sp. and Blastocystis plus Cyclospora sp. The haematic protozoa detected were Haemoproteus sp. (17%) and Plasmodium sp. (7.3%). Multiple parasitism by Haemoproteus sp. and Plasmodium sp. was detected in 1 sample of Gallus gallus. After each sampling, some of the affected animals were treated according to our results, and the corresponding programmes of prevention and control were designed. PMID- 19682801 TI - Clinical applications of rituximab in allogeneic stem cell transplantation: anti tumor and immunomodulatory effects. AB - Rituximab is a chimeric monoclonal antibody directed against CD20, a B-cell antigen expressed on B-cell lymphoma. It is widely used as single agent for the treatment of follicular lymphoma or in conjunction with other combination therapy as frontline or salvage therapy. Its efficiency in B-cell depletion has lead to other applications such as therapy for autoimmune diseases, GVHD and other immunologic complications in allogeneic stem cell transplantation. Clinical responses to rituximab unveiled the role of B-cells in the pathogenesis of these disorders. Attenuation of pathogenic antibody production partly explained the clinical response in patients with autoimmune disease and chronic GVHD, but other immune mechanisms might be operative as well. Expansion of regulatory T-cells (Tregs) following rituximab therapy indicated the interaction of T- and B-cells in chronic GVHD. Therefore, effort to maintain expansion of Tregs might be important for long-term control of these diseases. Other B-cell targeting strategy directing against B-cell-activating factor (BAFF) or its receptor could be considered in conjunction with rituximab. Recent CIBMTR data on reduced cumulative incidence of acute GVHD in patients who had prior rituximab also suggest the early role of B-cells in allogeneic transplantation, thus opening the opportunity for further immune modulation to prevent acute GVHD. PMID- 19682802 TI - Neonatal bacterial infection alters fever to live and simulated infections in adulthood. AB - Fever is a critical component of the host immune response to infection. An emerging literature demonstrates that experience with infectious organisms early in life, during the perinatal period, may permanently program immune responses later in life, including fever. We explored the influence of neonatal infection with Escherichia coli on fever responses to lipopolysaccharide (LPS) and E. coli in adulthood. Fever to a low dose of LPS in adulthood did not significantly differ as a consequence of early-life infection. Eight days after the LPS injection, the same group of rats received a high dose of live E. coli. This time, neonatally infected rats exhibited a markedly longer fever than controls. In a subsequent experiment, fever to a single high dose of E. coli without prior LPS in adulthood did not differ by group, suggesting that the previous difference was a lack of tolerance to the dual challenges in early-infected rats. Finally, both groups exhibited decreased tumor necrosis factor (TNF)-alpha and toll-like receptor (TLR) 4 production to dual LPS challenges in isolated splenocytes, whereas only rats infected as neonates exhibited increased cyclooxygenase-2 within the hypothalamus in response to adult infection, suggesting that early infection-induced changes in fever regulation may involve a change in central mechanisms. Taken together, these data indicate that early-life infection is associated with marked changes in host temperature regulation in adulthood. PMID- 19682803 TI - Acute exercise ameliorates reduced brain-derived neurotrophic factor in patients with panic disorder. AB - The neurotrophin brain-derived neurotrophic factor (BDNF) has been implicated in depression and anxiety. Antidepressants and exercise increase BDNF expression, and both have an antidepressant and anxiolytic activity. To further characterize the association of anxiety, BDNF and exercise, we studied panic disorder patients (n=12) and individually matched healthy control subjects (n=12) in a standardized exercise paradigm. Serum samples for BDNF analyses were taken before and after 30min of exercise (70 VO(2max)) or quiet rest. The two conditions were separated by 1 week and the order was randomized. Non-parametric statistical analyses were performed. There was a negative correlation of BDNF concentrations and subjective arousal at baseline (r=-0.42, p=0.006). Compared to healthy control subjects, patients with panic disorder had significantly reduced BDNF concentrations at baseline and 30min of exercise significantly increased BDNF concentrations only in these patients. Our results suggest that acute exercise ameliorates reduced BDNF concentrations in panic disorder patients and raise the question whether this is also found after long-term exercise training and if it is related to the therapeutic outcome. PMID- 19682804 TI - Rolling autogenetic dermis up to form a tube may be used as scaffold in tissue engineered blood vessels. AB - Coronary and peripheral artery bypass grafting are widely being used to deal with vascular deficiencies currently, and a man-made synthetic tube or autogenous arteries or veins are needed a lot. But one's autogenous arteries or veins are limited, and artificial graft substitute isn't yet available in clinical applications because of many disadvantages. Various polymeric materials have been used as scaffolds, but without satisfying results due to intimal hyperplasia and the rate of degradation. Autogenetic dermis, which has the advantages of resistance to immunogenicity, good biocompatibility, and appropriate mechanical and physiological properties, has gained our attention to use it as a scaffold for tissue-engineered blood vessels. What is more, autogenetic dermis can be harvested easily. So we postulate that autogenetic dermis rolled up to form a tube may be an ideal scaffold for tissue-engineered blood vessels. PMID- 19682805 TI - Clostridia as agents of zoonotic disease. AB - Clostridia are not normally considered to be zoonotic pathogens, although many species affect both humans and domestic animals. Three cases in which organisms occur, possibly via direct or indirect transmission, in both food animals and humans are considered here. Strains of Clostridium perfringens that produce enterotoxin (CPE) are typically transmitted to humans in contaminated, improperly handled foods. Pathogenesis is based upon action of CPE in the intestine, and disease is usually self-limiting. Infection of domestic animals by CPE-producing C. perfringens is uncommon. C. perfringens type C is best known as a pathogen of neonatal domestic animals, which acquire the infection from the dam. The course may be peracute, and prevention by vaccination of the dam is universally advocated. Humans consuming meat contaminated with type C may develop enteritis necroticans, with segmental hemorrhagic and necrotic jejunitis, which must usually be treated by bowel resection. Clostridium difficile is a pathogen of both humans and domestic animals. Examination of retail meats by bacteriologic culture has revealed genotypes of C. difficile that in many cases are identical to those from food animals and diseased humans. Transmission, food animals to foods to humans, has not been documented. PMID- 19682806 TI - Lectin binding patterns and carbohydrate mediation of sperm binding to llama oviductal cells in vitro. AB - Sperm binding to oviductal epithelium would be involved in sperm reservoir formation in the utero tubal junction (UTJ). Although in other mammals sperm oviduct interaction has been proved to be mediated by carbohydrate-recognition mechanisms, the factors implicated in the sperm adhesion to oviductal epithelium of llama are still unknown. In order to assess the role of carbohydrates present in the mucosa surface, we examined the distribution of glycoconjugates in the llama oviduct by confocal lectin-histochemistry. Mannosyl, glucosyl, N acetylglucosaminyl, galactosyl, N-acetylgalactosaminyl and sialic acid residues were detected in the oviductal mucose glycocalyx. By incubation of UTJ oviductal explants with LCA, DBA, UEA-1 or PNA lectin previous to co-culture with sperm, we observed a significant decrease in sperm binding only with LCA lectin. In the mucosa surface there were numerous d-glucosyl and D-manosyl residues, which were spotted by this lectin. Probably, this fact promotes the whole covering of the oviduct luminal surface by the sugar-lectin complex, preventing sperm access and adhesion of further residues. However, sperm incubation with mannose or glucose does not significantly prevent binding, which means that glucose and mannose would not be involved in a specific sperm-oviduct interaction. On the other hand, we observed a high reduction in sperm binding to UTJ explants with N acetylgalactosamine and galactose (p<0.001). Coincidentally, binding sites for N acetylgalactosamine-PAA-FITC conjugate were observed on the whole surface of the sperm, supporting the concept that llama sperm have lectin-like molecules in their surface, as is the case in other mammals. Probably, these lectin-like molecules, by means of N-acetylgalactosamine and galactose recognition, could link the sperm to the oviductal mucosa with the purpose of forming storing sites in the UTJ. Our results support the idea that more than one carbohydrate could participate in sperm reservoir formation in the llama UTJ oviductal segment. PMID- 19682807 TI - Intra-abdominal needle: medical malpractice? AB - This case relates to an intra-abdominal foreign body which was detected in a patient. The patient claimed that this was a needle which had been negligently left in his abdomen by a surgical team following an appendectomy which had been performed 13 years previously (in 1994). A radiograph was taken in 2007 and compared against the pre-operative radiograph which had been taken in 1994. This showed that the foreign body had been in the patient's body prior to the appendectomy being performed. The foreign body was extracted from the patient's liver through laparoscopic surgery in 2007. Examination of the fragments revealed that the foreign body was a sewing needle. Medical literature suggests that the needle may have accidentally been ingested by the patient. This is a rare case of a migrated swallowed foreign body (there are only 46 similar cases reported worldwide). It is of interest that the patient's immediate reaction was to assume that this was a case of medical malpractice. This report discusses issues relating to this case that were raised during the criminal proceedings relating to the medical negligence claim. PMID- 19682808 TI - Rapid oscillatory activity in delta brushes of premature and term neonatal EEG. AB - We compared frequency and power of neonatal EEG delta brush rapid oscillatory activity (ROA) using multiple band frequency analysis (MBFA) in three groups; pre term (PT, post-conceptional age 33-35.6 weeks, n=5); full-term (FT, 39.4-40.6 weeks, n=5) and pre-term or full-term with phenobarbital exposure (PB, n=5). Mean number of delta brushes analyzed was 29.4 (range 26-47) in PT, 20.8 (14-33) in FT and 20 (7-37) in PB. Mean frequency+/-standard deviation (s.d.) was 16.9+/-2.1 Hz (range 15-20 Hz) in PT, 17.3+/-1.9 Hz (15-20 Hz) in FT and 16.1+/-1.6 Hz (14-19 Hz) in PB. Mean power+/-s.d. was 22.9+/-6.2 microV(2) (range 16-39 microV(2)) in PT, 11.9+/-4.1 microV(2) (7-19 microV(2)) in FT and 17.1+/-6.2 microV(2) (9-26 microV(2)) in PB. Power was significantly higher in PT than FT (p<0.005). Power after merging PB into respective PT (PT', n=8) and FT (FT', n=7) groups, remained significantly higher in PT' (mean+/-s.d. 21.8+/-7.4 microV(2)) than FT' (11.4+/ 3.6 microV(2)) (p<0.05). We characterise ROA in delta brushes in maturing neonates using MBFA, which may provide additional information for assessing future seizure recurrence and epilepsy risk. PMID- 19682809 TI - Magnetic resonance imaging at first episode in pediatric multiple sclerosis retrospective evaluation according to KIDMUS and lesion dissemination in space criteria. AB - BACKGROUND: Several diagnostic imaging criteria are being described and examined in pediatric multiple sclerosis (MS). Compared to adults, children are more likely to experience acute or relapsing demyelinating episodes of various etiologies which show similar clinical and magnetic resonance imaging (MRI) findings. AIM: To investigate the fulfillment of MRI diagnostic criteria at initial episode in pediatric MS. METHODS: We reviewed our series of children and adolescents with the final diagnosis of clinically definite MS and applied the McDonald dissemination in space (DIS) and KIDMUS criteria to their initial MRI scans. RESULTS: Thirty patients (17 girls, 13 boys), most with brainstem dysfunction and polysymptomatic presentation, were included in the study. Twenty five (83.3%) patients fulfilled both McDonald and KIDMUS criteria. Patients who did not meet any McDonald DIS criteria did not meet KIDMUS criteria either. Only one patient met the McDonald criteria but not the KIDMUS criteria because of the absence of lesions perpendicular to corpus callosum. CONCLUSIONS: Our results show 5/30 (16.6%) of MS patients may not present the diagnostic MRI features initially. The variable sensitivity observed for the current MRI criteria in different series can be due to referral biases, differences between populations and length of follow-up, and the definition of MS patients by two attacks only. PMID- 19682810 TI - [Vascular medicine: Comments on the future of the speciality and other timely topics]. PMID- 19682811 TI - [Anorectal disease: past, present, future]. AB - Over the last decade, therapeutic approaches of anorectal disorders have been profoundly modified by new drugs, new procedures and functional considerations. In fact, the primary goals of these procedures emphasize minimal invasive approaches. Less functional postoperative complaints are often preferred over a radical efficacy. As compared to haemorrhoidectomy, haemorrhoidopexy procedure is today advocated to reduce postoperative care and complaints. As compared to lateral sphincterotomy, nitrates and botulinum toxin represent a second line therapy of chronic anal fissure to avoid faecal incontinence. As compared to fistulotmy, both glue and plug may be used to treat a high tract fistulae for the same reasons. PMID- 19682812 TI - [Gastroduodenal ulcer before and after Helicobacter pylori]. AB - Before the discovery of Helicobacter pylori, ulcer disease was considered as the result of a conflict between gastric acid and pepsin, on one side, and protection afforded by gastric mucosal barrier, on the other side. The discovery of H. pylori by Marshall and Warren in 1982 overthrew this conception and revealed ulcer disease mainly as an infectious disease. H. pylori eradication with an appropriate triple therapy is now considered as the gold standard treatment of gastroduodenal ulcer. The pathogenic role of H. pylori lies far beyond ulcer disease since H. pylori is looked as involved in nonulcer dyspepsia, nonsteroidal anti-inflammatory drug ulcers, gastric cancer, MALT lymphoma and, eventually, oesophageal adenocarcinoma, and nondigestive diseases as cardiovascular diseases. The pandemic nature of the H. pylori infection, particularly within developing countries, combined with emerging resistances to antibiotics make the development of a vaccine a public health necessity. The relationships between the human host and the bacterium remains mostly unknown, some of which could be beneficial. PMID- 19682813 TI - [Irritable bowel syndrome: from the gut to the brain-gut]. AB - Irritable bowel syndrome is not only a digestive motor disorder. It is a multifactorial disease for which many data have highlighted the pathophysiological importance of visceral hypersensitivity in the onset of symptoms, particularly abdominal pain. Hypersensitivity is due either to an afferent neurons disfunction at the enteric nervous system level, either to an abnormal brain-gut axis processing of sensory or nociceptive inputs arising from the gut, at the spinal or supraspinal level. Disturbances of the autonomic nervous system occur in IBS as a consequence of this brain-gut axis dysfunction. Neurological abnormalities may be triggered by inflammation, mast cell dysfunction or increased intestinal permeability while the neuro-immune consequences of stress (mainly chronic) play a major role in the genesis and the maintenance of irritable bowel syndrome. The role of emotions and mood disturbances cannot be omitted in the interpretation the central processing of digestive sensory inputs. Neurosciences, in particular brain imaging techniques, have contributed to this better understanding of irritable bowel syndrome pathophysiology. It is likely to play a major role in the future to improve our knowledge of the brain-gut axis function (mechanisms, neurotransmitters and receptors involved both in normal and pathological conditions). This knowledge is crucial because of the need for updated treatment strategies and new pharmacological and/or cognitive or behavioral therapies. PMID- 19682815 TI - Quantitative computed tomography. AB - Quantitative computed tomography (QCT) was introduced in the mid 1970s. The technique is most commonly applied to 2D slices in the lumbar spine to measure trabecular bone mineral density (BMD; mg/cm(3)). Although not as widely utilized as dual-energy X-ray absortiometry (DXA) QCT has some advantages when studying the skeleton (separate measures of cortical and trabecular BMD; measurement of volumetric, as opposed to 'areal' DXA-BMDa, so not size dependent; geometric and structural parameters obtained which contribute to bone strength). A limitation is that the World Health Organisation (WHO) definition of osteoporosis in terms of bone densitometry (T score -2.5 or below using DXA) is not applicable. QCT can be performed on conventional body CT scanners, or at peripheral sites (radius, tibia) using smaller, less expensive dedicated peripheral CT scanners (pQCT). Although the ionising radiation dose of spinal QCT is higher than for DXA, the dose compares favorably with those of other radiographic procedures (spinal radiographs) performed in patients suspected of having osteoporosis. The radiation dose from peripheral QCT scanners is negligible. Technical developments in CT (spiral multi-detector CT; improved spatial resolution) allow rapid acquisition of 3D volume images which enable QCT to be applied to the clinically important site of the proximal femur, more sophisticated analysis of cortical and trabecular bone, the imaging of trabecular structure and the application of finite element analysis (FEA). Such research studies contribute importantly to the understanding of bone growth and development, the effect of disease and treatment on the skeleton and the biomechanics of bone strength and fracture. PMID- 19682814 TI - [Celiac disease in 2009: a future without gluten-free diet?]. AB - Celiac disease is an enteropathy related to autoimmune diseases induced by gluten in genetically predisposed individuals. Its prevalence is of 1% in Europe and United States. Its clinical presentation is extremely various and diagnosis relies on the detection of specific serum antibodies and on the demonstration of intestinal villous atrophy. Treatment relies on a life-long gluten free diet which prevents bone, autoimmune and malignant complications. The keystone of its pathogenesis is the interaction of gliadin peptides with HLA DQ2/8 molecules, the main genetic risk factor, which induces the activation of CD4+ T-cells in the lamina propria. Yet, complementary mechanisms are necessary to provoke the loss of tolerance to gluten which involves the cytokine IL-15 responsible of the activation/expansion of intraepithelial lymphocytes, a hallmark of the origin of the severe lymphomatous complications. The burden of the gluten-free diet leads to a strong demand for alternative treatments. Numerous strategies have been identified to prevent the recognition of gliadin peptides by the immune system. Their efficiency and safety remained to be evaluated, the most attainable strategy today being oral therapy by enzymes able to eliminate gluten immunogenicity. PMID- 19682816 TI - Genotypic and phenotypic characterization of rhizobia that nodulate snap bean (Phaseolus vulgaris L.) in Egyptian soils. AB - Snap bean fields in 12 of the 25 governorates of Egypt were surveyed to determine the distribution and taxonomy of snap bean-nodulating rhizobia. Nodulation rates in the field were very low, indicating that Egyptian soils do not have sufficient numbers of snap bean-compatible Rhizobium spp. A total of 87 rhizobial isolates were assayed on the most commonly grown cultivars in order to identify the most effective strains. The five most effective isolates (R11, R13, R28, R49 and R52) were fast-growing and utilized a wide range of carbon and nitrogen sources. A phylogenetic assignment of these strains by analysis of the 16S ribosomal RNA gene suggested that all fell within the Rhizobium etli-Rhizobium leguminosarum group. Strains R11, R49 and R52 all clustered with other identified R. etli strains, while strains R13 and R28 were more distinct. The distinctness of R13 and R28 was supported by physiological characteristics, such as their ability to utilize citrate, erythritol, dulcitol and lactate. Strains R13 and R28 also yielded the highest plant nitrogen content of all isolates. The highly effective strains isolated in this study, in particular strains R13 and R28, are promising candidates for improving crop yields. The data also suggested that these two strains represented a novel sub-group within the R. etli-R. leguminosarum group. As snap bean is a crop of great economic value to Egypt, the identification of highly effective rhizobial strains adapted to Egyptian soils, such as strains R13 and R28, is of great interest. PMID- 19682817 TI - Theoretical problems associated with the use of acetic anhydride as a co-solvent for the non-aqueous titration of hydrohalides of organic bases and quaternary ammonium salts. AB - A potentiometric titration study of organic base hydrohalides and quaternary ammonium salts using perchloric acid as the titrant and a mixture of acetic anhydride and acetic acid as the solvent was carried out and the titration mixture was analysed by NMR in order to clarify the chemistry of the reactions involved. It was found that in contrast to the general belief the formation of acetyl halides and titratable free acetate ion does not take place prior to the titration but NMR spectra proved the formation of acetyl halides in the course of the titration. This observation and the fact that the shape of the titration curves depends on the nature of the hydrohaloic acid bound to the base or of the anion in the quaternary ammonium salts led to the conclusion that the titrating agent is acetyl perchlorate formed in situ during the titration. Equations of the reactions involved in the titration process are shown in the paper. PMID- 19682818 TI - EuroCMR (European Cardiovascular Magnetic Resonance) registry: results of the German pilot phase. AB - OBJECTIVES: During its German pilot phase, the EuroCMR (European Cardiovascular Magnetic Resonance) registry sought to evaluate indications, image quality, safety, and impact on patient management of routine CMR. BACKGROUND: CMR has a broad range of applications and is increasingly used in clinical practice. METHODS: This was a multicenter registry with consecutive enrollment of patients in 20 German centers. RESULTS: A total of 11,040 consecutive patients were enrolled. Eighty-eight percent of patients received gadolinium-based contrast agents. Twenty-one percent underwent adenosine perfusion, and 11% high-dose dobutamine-stress CMR. The most important indications were workup of myocarditis/cardiomyopathies (32%), risk stratification in suspected coronary artery disease/ischemia (31%), as well as assessment of viability (15%). Image quality was good in 90.1%, moderate in 8.1%, and inadequate in 1.8% of cases. Severe complications occurred in 0.05%, and were all associated with stress testing. No patient died during or due to CMR. In nearly two-thirds of patients, CMR findings impacted patient management. Importantly, in 16% of cases the final diagnosis based on CMR was different from the diagnosis before CMR, leading to a complete change in management. In more than 86% of cases, CMR was capable of satisfying all imaging needs so that no further imaging was required. CONCLUSIONS: CMR is frequently performed in clinical practice in many participating centers. The most important indications are workup of myocarditis/cardiomyopathies, risk stratification in suspected coronary artery disease/ischemia, and assessment of viability. CMR imaging as used in the centers of the pilot registry is a safe procedure, has diagnostic image quality in 98% of cases, and its results have strong impact on patient management. PMID- 19682821 TI - Acute pulmonary edema and cardiac failure due to zinc phosphide ingestion. PMID- 19682819 TI - Effects of angiotensin-converting enzyme inhibitor therapy on clinical outcome in patients undergoing coronary artery bypass grafting. AB - OBJECTIVES: This study evaluates the effect of pre-operative angiotensin converting enzyme inhibitor (ACEI) therapy on early clinical outcomes after coronary artery bypass grafting (CABG). BACKGROUND: Therapy with ACEIs has been shown to reduce the rate of mortality and prevent cardiovascular events in patients with coronary artery disease. However, their pre-operative use in patients undergoing CABG is still controversial. METHODS: A retrospective, observational, cohort study was undertaken of prospectively collected data on 10,023 consecutive patients undergoing isolated CABG between April 1996 and May 2008. Of these, 3,052 patients receiving pre-operative ACEI were matched to a control group by propensity score analysis. RESULTS: Overall rate of mortality was 1%. Pre-operative ACEI therapy was associated with a doubling in the risk of death (1.3% vs. 0.7%; odds ratio [OR]: 2.00, 95% confidence interval [CI]: 1.17 to 3.42; p = 0.013). There was also a significant difference between the ACEI and control group in the risk of post-operative renal dysfunction (PRD) (7.1% vs. 5.4%; OR: 1.36, 95% CI: 1.1 to 1.67; p = 0.006), atrial fibrillation (AF) (25% vs. 20%; OR: 1.34, 95% CI: 1.18 to 1.51; p < 0.0001), and increased use of inotropic support (45.9% vs. 41.1%; OR: 1.22, 95% CI: 1.1 to 1.36; p < 0.0001). In a multivariate analysis, pre-operative ACEI treatment was an independent predictor of mortality (p = 0.04), PRD (p = 0.0002), use of inotropic drugs (p < 0.0001), and AF (p < 0.0001). CONCLUSIONS: Pre-operative therapy with ACEI is associated with an increased risk of mortality, use of inotropic support, PRD, and new onset of post-operative AF. PMID- 19682820 TI - High-sensitive troponin testing and the "runner's syndrome". PMID- 19682822 TI - Resident involvement in civilian tactical emergency medicine. AB - BACKGROUND: Tactical emergency medicine services (TEMS) has emerged as a specialized niche within the field of emergency medicine. With increasing demand for physician participation in civilian tactical teams, there will be efforts by residents to become involved at earlier points in their clinical training. OBJECTIVES: This article discusses resident involvement with a civilian TEMS unit and provides five maxims for emergency physicians to better understand the difference between working in the emergency department or with emergency medical services vs. in a TEMS environment. DISCUSSION: Differences between TEMS and other trauma life support models, institutional and political barriers likely to be encountered by the resident, the value of preventive medicine and the role of the physician in long-term tactical operations, opportunities for subspecialty growth, and the role of operational security are all discussed in detail. CONCLUSION: Tactical emergency medicine is a specialty that utilizes the full array of the emergency physician's skill set. It is also a field that is ripe for continued expansion, but the resident looking to become involved with a team should be aware of the requirements necessary to do so and the obstacles likely to be encountered along the way. PMID- 19682823 TI - Bleeding scrotal arteriovenous malformation. AB - BACKGROUND: Arteriovenous malformation (AVM) of the scrotum has, to our knowledge, been reported fewer than 10 times in the medical literature. It may be of congenital or post-traumatic etiology and has been reported to present with spontaneous bleeding. CASE REPORT: A case of a spontaneously bleeding scrotal AVM that developed after remote trauma is presented. The differential diagnosis of a scrotal vascular lesion includes varicocele, hemangioma, lymphangioma, and AVM. Although ultrasound and magnetic resonance imaging play an important role in the evaluation, angiography is essential to fully delineate the feeder vessels, vascular takeoffs, and draining veins. CONCLUSION: Scrotal AVMs are rare vascular scrotal lesions that may present with scrotal enlargement and a bruit. Definitive therapy should be planned in concert with Interventional Radiology and surgical consultants. PMID- 19682824 TI - Persistent ventricular fibrillation during therapeutic hypothermia and prolonged high-dose vasopressor therapy: case report. AB - BACKGROUND: Recent emphasis on high quality prehospital cardiopulmonary resuscitation has resulted in more out-of-hospital cardiac arrest victims surviving to the emergency department. As such, standardized in-hospital post cardiac arrest care is necessary to assure optimal neurological recovery. Although therapeutic hypothermia has arisen as a key component in the post cardiac arrest care paradigm, its interaction with other therapies remains poorly defined. OBJECTIVE: The purpose of this communication is to demonstrate a potential interaction between therapeutic hypothermia and routinely administered resuscitation medications. CASE REPORT: We present a case of idiopathic ventricular fibrillation in a previously healthy 36-year-old man who developed persistent ventricular fibrillation in the setting of mild therapeutic hypothermia and high doses of routine resuscitation medications. CONCLUSION: This case illustrates the importance of understanding the potential interaction between therapeutic hypothermia and resuscitation medications along with the need for a systematic and standardized, multi-disciplinary approach to post-cardiac arrest care. PMID- 19682826 TI - A case of catamenial pneumothorax with diaphragmatic fenestrations. AB - BACKGROUND: Considerable controversy exists with regards to the physiopathogenesis of catamenial pneumothorax. The rarity of catamenial pneumothorax makes understanding of its pathophysiology and verification of etiological mechanisms difficult. OBJECTIVE: To contribute evidence to the knowledge base on the pathogenesis of catamenial pneumothorax. CASE REPORT: We describe a case of catamenial pneumothorax with images that substantiate the pore hypothesis as a cause of recurrence of air in the pleural cavity in this patient. CONCLUSION: Our case report contributes evidence that transperitoneal migration of endometrial implants may occur through diaphragmatic fenestrations. Surgical options may be more viable to prevent recurrent pneumothoraces in such patients. PMID- 19682827 TI - Transverse sinus thrombosis after internal jugular vein ligation. AB - BACKGROUND: Cerebral vein and dural sinus thrombosis is a rare condition with a wide range of causes and a highly variable presentation. It can lead to significant morbidity, but scant literature is available describing diagnosis and treatment when this occurs after ligation of the internal jugular vein. OBJECTIVES: To discuss potential risk factors for cerebral vein and dural sinus thrombosis after ligation of the internal jugular vein, and present current options for diagnosis and treatment. CASE REPORT: A 23-year-old male construction worker was brought to the Emergency Department by Emergency Medical Services after sustaining a severe neck laceration from a hand-held grinder. He was treated with ligation of the left internal jugular vein, but subsequently developed severe headaches and symptoms of increased intracranial pressure. A magnetic resonance venogram of the head revealed a left transverse sinus thrombosis requiring treatment with anticoagulation. The placement of a lumboperitoneal shunt was ultimately needed for relief of his symptoms. CONCLUSIONS: Early diagnosis and aggressive therapeutic interventions are critical to prevent further morbidity in patients who develop cerebral vein and dural sinus thrombosis after ligation of the internal jugular vein. PMID- 19682828 TI - Emergency department workload increase: dependence on primary care? AB - BACKGROUND: Increasing demand for emergency care and crowded emergency departments (EDs) lead some planners to conclude that inconvenient primary care scheduling increases the number of "unnecessary" ED visits. The reasons that the planners argue for more primary care are: to increase funding for primary care; the unfounded notion that it is less expensive to see a primary care physician (PCP) than an Emergency Physician; and the impractical goal that the ED should be used only by intellectually interesting life- or limb-threatened patients or "true emergencies." OBJECTIVE: To explore the rates of patient-reported access to primary care in ambulatory presentations to a rural tertiary care ED. METHODS: An observational study was performed in which an anonymous survey was given to a convenience sample of patients who presented by walking into the ED. RESULTS: Overall, 70.4% (686/975) of respondents stated that they had a PCP, and 38.1 % (252/661) of the sample had attempted to contact their physicians before presenting to the ED. Of the group who attempted to contact their physicians, 62.8% (130) were neither spoken to nor seen by any doctor. These rates did not change by time of presentation or by day of the week. CONCLUSION: The results suggest that it is neither a lack of primary care, nor the time of day or night that drives patients to come to the ED. PMID- 19682829 TI - De novo cerebral arteriovenous malformation: Pink Floyd's song "Brick in the Wall" as a warning sign. AB - BACKGROUND: Arteriovenous malformations are shunts between an artery and the venous system that lie within a nidus without an intervening capillary bed. These lesions are thought to be congenital, but recent reports have challenged this assumption. CASE REPORT: A 50-year-old man presented to the emergency department with a generalized tonic-clonic seizure. Before the onset of his seizure, he experienced a vivid auditory hallucination of his favorite song by the band Pink Floyd, "A Brick in the Wall." He had been diagnosed with epilepsy 25 years previously. On presentation, his neurological examination was normal, but a computed tomography scan of the brain revealed a large arteriovenous malformation (AVM) occupying the left temporal lobe. Upon more detailed questioning, he recalled that a brain angiogram had been performed 25 years before and was reported to be normal. Neurosurgery was not performed in view of the size of the malformation. The patient is being followed-up as an outpatient. CONCLUSION: AVMs may arise de novo and then spontaneously become symptomatic. Cerebral de novo AVM should be considered in the differential diagnosis in patients with complex auditory musical hallucinations or any new neuropsychiatric symptoms. PMID- 19682830 TI - Patient empowerment: myths and misconceptions. AB - OBJECTIVE: The purpose of this article is to clarify the concept of empowerment and to correct common misconceptions about its use in diabetes care and education. METHODS: The patient empowerment approach is well suited to helping patients make self-selected changes related to weight, nutrition, and physical activity. Although the concept of patient empowerment has become an integral part of diabetes education, an accurate understanding and authentic application of empowerment has not occurred as readily. The empowerment approach is clarified and common misconceptions have been corrected. RESULTS: Embracing empowerment means making a paradigm shift that is often difficult because the traditional approach to care is embedded in the training and socialization of most health care professionals (HCPs). CONCLUSION: Unlike the traditional approach, empowerment is not something one does to patients. Rather, empowerment begins when HCPs acknowledge that patient are in control of their daily diabetes care. Empowerment occurs when the HCPs goal is to increase the capacity of patients to think critically and make autonomous, informed decisions. Empowerment also occurs when patients are actually making autonomous, informed decisions about their diabetes self-management. PRACTICE IMPLICATIONS: Clarity about all aspects of the empowerment approach is essential if it is to be used effectively. PMID- 19682831 TI - Perceptions of couple HIV counseling and testing in Botswana: a stakeholder analysis. AB - OBJECTIVE: To explore stakeholder's perceptions of Couples HIV Counseling and Testing (CHCT) as opposed to individual testing and potential couples' preferences for CHCT promotion and service provision. METHODS: Study was conducted as formative research for a phase III clinical trial of Herpes (HSV-2) suppression to prevent HIV transmission among HIV discordant couples. We used non probability purposive sampling and snowballing techniques to identify study participants. Data were collected using key informant interviews and focus group discussions. Systematic textual data analysis was used. Two independent coders coded and compared their codes for agreement. Data was categorized by emerging themes. RESULTS: The general themes from both key informant interviews and focus group discussions were a preference for CHCT as opposed to individual counseling in HIV prevention and the need for a client-centered approach to promotion and provision of couple HIV testing services. CONCLUSION: CHCT is important in HIV prevention and should be integrated in existing HIV testing programs. The study also demonstrates the challenges of HIV status disclosure and discordance among sexual partners who test as individuals. PRACTICE IMPLICATIONS: Current low HIV status disclosure rates imply that reducing HIV incidence rates will require integrating CHCT into current testing programs. Increasing CHCT uptake however, requires improving access, training providers and addressing social, cultural, political and logistical barriers. PMID- 19682833 TI - Radiation therapy in the treatment of desmoid tumours reduces surgical indications. AB - BACKGROUND: While several modalities have been proposed for the treatment of desmoid tumour/aggressive fibromatosis, high local recurrence rates have been reported. We present a retrospective study of including patients treated with radiation therapy, some of them in combination with surgical resection. PATIENTS AND METHODS: Thirty-four consecutive patients were included (mean age 40+/-16 years, 9 male). Complete follow-up was available in 31 patients (51+/-36 months). Seventeen patients (50%) were treated with radiation therapy alone, 17 patients with radiation therapy and surgery. Radiation therapy (external beam) was applied in most cases to a total dose of 50.4 Gy in 28 fractions. The lesion was located in the upper extremity in 11 patients, in the lower extremity in 14 cases and on the trunk in 9 cases. RESULTS: Overall recurrence/progression free survival was 88.5% at 5 years and 77.5% at 10 years. Recurrence free survival of the subset of patients undergoing combined treatment with radiation therapy and surgical resection was 83.6% at 5 years and 10 years. In patients who did not receive surgery but only radiation therapy, MRI showed a complete response in 20%, a partial response in 20%, and stable disease in 53% of cases. In this subset, two third of patient had a metabolic response to radiotherapy (i.e. decrease uptake on the thallium-210 scan after radiotherapy compared to pre-therapy levels). CONCLUSION: Low recurrence rates can be achieved with the use of radiation therapy alone in selected cases. Patients with a metabolic response (decrease) to radiotherapy may be treated with a non-surgical approach. Surgery might be considered in patients with a poor metabolic response to radiotherapy. PMID- 19682834 TI - [Headaches in a 21-year-old man with Goodpasture disease]. AB - The case of a 21-year old man who died due to an intracranial thrombosis just after diagnosis of Goodpasture's disease, is reported. Discussion deals with the putative mechanisms, which could be responsible for the thrombosis. PMID- 19682835 TI - [A very extensive necrotizing fasciitis]. AB - Necrotizing fasciitis is a severe skin infection. Fluidized bed may be indicated to improve healing. We report a 36-year-old woman case, who developed an important skin emphysema on a fluidized bed that may have worsen the situation. PMID- 19682836 TI - [Exchanges between patients on the Internet]. AB - Internet is an important place for debates between patients, whose active discussions allow the patient to appropriate medical information and to build up their expertise. Lists and forums are regulated and organised areas; discussions about common rules take up a large place in the discussion. On discussion lists, we observe that speech is unequally distributed: 10% of the most talkative speakers send between 50 and 70% of all the messages; we also observe the temporal stability of the groups and the weak prevalence of anonymity which is made possible because the group is not open freely to the general public. Experimented speakers dominate discussions on lists as on forums. 4 speakers' profiles can be distinguished: the adviser, the agitator, the hyperactive person, and the charismatic leader. The tone of a list is defined, among others, by the distribution of those profiles. A systematic study of the contents of discussion lists reveals six main topics within the debates on health: collective, personal and interpersonal, medicine, patient / user, information, and phatic. We can distinguish various kinds of lists: lists focused on the articulation between patients experiences and medical expertise, lists mainly concerned by the exchange of personal experiences, lists organized around the sharing of medical information, and finally lists dedicated to mutual support. In collaboratively contributing to the production of health information, patients involved in these discussions can open the way to enrichment of medical knowledge. PMID- 19682838 TI - Detection of locking bolt loosening in the stem-condyle junction of a modular femoral stem in revision total knee arthroplasty. AB - We report one case of loosening of the locking bolt in the stem-condyle junction of a constrained modular femoral component in revision total knee arthroplasty. Early detection of loosening of the locking bolt was possible using reconstructed view of computerized tomography scans before complete disengagement and radiographic loosening of the stemmed femoral component. When using the Total Condyle III femoral component with a stem extension, surgeons should keep in mind that stress concentration at the stem-condyle junction can result in failure of the locking bolt although it is rare and that loosening or disengagement of the locking bolt may be a sign of a loose stemmed femoral component. PMID- 19682837 TI - Hip resurfacing arthroplasty for ankylosing spondylitis. AB - We compared the results of the metal-on-metal hip resurfacing with the ceramic-on ceramic total hip arthroplasty (THA) in 2 groups. The preoperative and postoperative ranges of motion (ROMs) were recorded. At the latest follow-up, both of the groups make satisfactory clinical and radiographic results. There was no significant difference in Harris hip score of the 2 groups, but the ROM of the hips in hip resurfacing group was significantly wider than THA group (P < .01). Hip resurfacing has better ROM improvement than THA, with the same pain relief. Its high stability and low dislocation rate allow patients to do early function exercises, which is important for ankylosing spondylitis (AS) patients to avoid reankylosis. Hip resurfacing may be a reasonable option for young AS population. PMID- 19682839 TI - First series of Exeter small stem primary total hip arthroplasty minimum 5 years of follow-up. AB - We carried out a prospective study of 47 Exeter (Stryker Inc, Warsaw, Ind) small stem total hip arthroplasty in 42 patients with an average age of 58 years and a mean follow-up of 8.5 years. The Oxford hip score improved from a preoperative mean of 47 to 17 at last follow-up. More than 87% patients had excellent or good Harris hip scores, and 90% were able to walk with little or no pain. Stem subsidence within the cement mantle was observed in 26% of cases, and none showed evidence of aseptic loosening or implant failure. Two stems were removed due to infection. The survival rate of this implant was 95.7% at 10 years. This first series of Exeter small stem showed excellent medium-term results, comparable to its larger counterparts. PMID- 19682840 TI - Risk factors and treatment outcome in scuba divers with spinal cord decompression sickness. AB - PURPOSE: This study was designed to determine the recompression strategy and the potential risk factors associated with the development of severe diving-related spinal cord decompression sickness (DCS). MATERIAL AND METHODS: Sixty-three injured recreational divers (52 men and 11 women; 46 +/- 12 years) presenting with symptoms of spinal involvement were retrospectively included. Diving information, symptom latency after dive completion, and time interval between symptom onset and hyperbaric treatment were studied. The severity of spinal cord DCS was rated numerically for both the acute event and 1-month later. Initial recompression treatment at 2.8 atmosphere absolute (ATA) with 100% oxygen breathing or deeper recompression at 4 atmosphere absolute with nitrogen-oxygen or helium-oxygen breathing mixture was also noted. RESULTS: Twenty-one divers (33%) had incomplete resolution after 1 month. The clinical severity at presentation was the only independent predictor of poor outcome (odd ratio, 2.68; P < .033). Time to treatment did not influence the recovery with a similar median delay (3 hours) between the divers with or without long-term sequelae. Choice of recompression procedure was not also a determinant factor for treatment outcome. CONCLUSION: The initial clinical course before treatment is a major prognostic factor of spinal cord DCS. Delay to recompression less than 3 hours and use of deep treatment tables did not improve outcome in DCS divers. PMID- 19682841 TI - Severe childhood injuries and poisoning in a densely populated city: where do they occur and what type? AB - AIM: The aim of this study was to review the patterns of severe childhood injuries and poisoning necessitating pediatric intensive care in a regional trauma center. METHODS: We reviewed discharge data of all children who were hospitalized for severe injuries and poisoning at the pediatric intensive care unit of a teaching hospital between October 2002 and December 2008. RESULTS: There were 86 patients (males/females, 2:1). Road traffic injuries (n = 19), falls (n = 17), and scalds (n = 13) were the 3 leading categories and accounted for 57% of these pediatric intensive care unit admissions. Injuries more commonly occurred indoor (63%), and victims of indoor accidents were younger (median age, 2.6 vs 8.4 years; P < .0001), with scalds, poisoning, and foreign body aspiration being predominant, whereas road traffic accidents predominated in outdoor accidents. As a risk factor, premorbid neurodevelopmental conditions such as mental retardation, convulsion disorder, or cerebral palsy were only present in indoor accidents. Children in outdoor injuries were generally healthy. Both groups were associated with significant morbidity (mechanical ventilation in 60%, inotrope use in 20%, anticonvulsants in 24%, and neurological/neurosurgical supports in 49%). Comparing the 3 most common categories of patients, there were significant differences in the median age, requirement of neurological/neurosurgical supports, and median hospital stay. Although not requiring neurological/neurosurgical supports, scalds injuries involved the youngest age group and required the longest hospital stay. CONCLUSIONS: The causes of severe childhood injuries are heterogeneous. Cardiopulmonary or neurological/neurosurgical supports are often required. These injuries more commonly occur indoor and involve toddlers with underlying neurodevelopmental conditions. These findings have important implications and serve to heighten public awareness especially on home safety measures in the prevention of childhood accidents. PMID- 19682842 TI - Contamination of 0.2-micrometer infusion filters by N,N-dimethylacrylamide. AB - PURPOSE: Infusion filters, 0.2 mum, are commonly used in intensive care units as in-line filters to minimize particle and microbiological burden on patients. These filters usually contain either a positively charged or an uncharged membrane. The aim of the present study was to identify and to quantify an additive causing an unexpected maximum at 234.5 nm in the ultraviolet spectrum of a filtered drug solution using a filter. MATERIALS AND METHODS: For identification and quantification of the substance, water for injection was used as eluent. Measurements were done by mass spectrometry and ultraviolet spectroscopy. RESULTS: The unexpected additive in the filter was found to be N,N dimethylacrylamide. The eluate of 2 filter batches had a mean N,N dimethylacrylamide concentration of 3.9 and 2.5 microg/mL, respectively. Further investigations showed that after infusion breaks of different duration, the N,N dimethylacrylamide concentration at the beginning of the next infusion cycle reaches a higher level than that at the end of the preceding infusion cycle. CONCLUSIONS: The type of filter examined in this study is primarily used in premature infants, newborns, and infants. These patients are vulnerable to neurotoxic substances having a long-term effect. Therefore, N,N dimethylacrylamide should be completely removed from the final filter product. PMID- 19682843 TI - Usefulness of intra-abdominal pressure in a predominantly medical intensive care unit. AB - BACKGROUND: The deleterious effects of elevated intra-abdominal pressure (IAP) have been known for more than a century. The proposed objectives were to measure changes in IAP and analyze increase-related factors and complications and whether high IAP and its persistence are related to complications and mortality in a predominantly medical intensive care unit. METHODS: Over a 1-year period, we conducted a prospective cohort study in which IAP was measured using the bladder method. Hospitalization time, demographic variables, diagnosis on admission, APACHE II score, and clinical complications were recorded. RESULTS: A total of 130 patients were studied. Overall mean IAP was 12.3 mm Hg (standard deviation [SD], 3.79; 95% confidence interval [CI], 11.7-13), and on the first day, 12.68 mm Hg (SD, 5.32; 95% CI, 11.8-13.6); maximum IAP was 16.4 mm Hg (SD, 4.6; 95% CI, 15.6-17.2). A positive correlation was found between IAP, APACHE (Acute Physiology And Chronic Health Evaluation) II, and age. Higher IAP values were independently associated with higher age, prolonged activated partial thromboplastin time, need for dialysis, and intolerance to enteral feeding. The value showing the best sensitivity and specificity in predicting mortality was persistence of IAP 20 mm Hg or greater for 4 days or more. The number of days with IAP 20 mm Hg or greater was a factor associated with a higher risk of death (odds ratio, 2.3). Patients who died showed a tendency to increased IAP. CONCLUSION: In this study, a threshold IAP of 20 mm Hg and its permanence over time were the best predictive factors of complications and mortality. Among other relationships, we also observed that older patients had higher IAP. High IAP was a cause of intolerance to enteral nutrition. PMID- 19682844 TI - A new dosing protocol reduces dexmedetomidine-associated hypotension in critically ill surgical patients. AB - BACKGROUND: Although no ideal sedative exists, dexmedetomidine is unique because it produces sedation and analgesia without decreasing the respiratory drive. Hemodynamic responses to dexmedetomidine are variable and dependent on the patient population. Our initial experience was associated with an unacceptable incidence of hypotension and bradycardia. We evaluated occurrence of hypotension and bradycardia in critically ill surgical patients receiving dexmedetomidine before and after implementation of a dosing protocol. METHODS: This is a retrospective chart review of all admissions to a university medical center based, 44-bed surgical intensive care unit pre and post protocol implementation. RESULTS: Forty-four patients received dexmedetomidine including 19 historic controls and 25 dosed via protocol. Both groups had comparable demographics and initial and maximum dosages of dexmedetomidine. Use of the dosing protocol resulted in fewer dosage changes (mean +/- standard deviation, 4.8 +/- 3.8 compared to 7.8 +/- 3.9; P = .014) and fewer episodes of hypotension (16% vs 68.4%; P = .0006) but did not influence bradycardic episodes (20% vs 15.5%; P > .99). CONCLUSION: We found that use of a protocol that increases the time interval between dosage adjustments may reduce dexmedetomidine-associated hypotension. PMID- 19682845 TI - Effects of polygeline and hydroxyethyl starch solutions on liver functions assessed with LIMON in hypovolemic patients. AB - BACKGROUND: Hypovolemia is a common clinical entity in critical patients, and adequate volume replacement therapy seems to be essential for maintaining tissue perfusion. However, it is still uncertain which solution is most appropriate for fluid resuscitation. OBJECTIVE: The aim of this study was to investigate the effects of fluid resuscitation with 3.5% polygeline versus 6% hydroxyethyl starch solutions on hemodynamic functions and liver functions assessed with a noninvasive liver function monitoring system (LIMON) in hypovolemic patients. DESIGN: This study is a prospective randomized clinical trial. MEASUREMENTS AND RESULTS: Thirty hypovolemic patients (intrathoracic blood volume index, <850 mL/m(2)) were randomized into hydroxyethyl starch (mean molecular weight, 130,000 Da) and polygeline (mean molecular weight, 30,000 Da) groups (15 patients each). Indocyanine green plasma disappearance elimination (ICG-PDR) were conducted concurrently using LIMON. A dose of 0.3 mg/kg ICG was given through a cubital fossa vein as a bolus. For fluid resuscitation, 500 mL of colloid was given to the patients. Repeated hemodynamic and ICG-PDR measurements were done at baseline, after infusion, and then at 30 minutes after infusion. RESULTS: Intrathoracic blood volume index and systolic, diastolic, and mean blood pressures increased significantly after infusion and remained elevated for 30 minutes after infusion, but there was no significant difference between the 2 groups. Indocyanine green plasma disappearance elimination values were similar in both groups with no significant difference between the two. CONCLUSION: Increasing intrathoracic blood volume index and hemodynamic variables by fluid loading is not associated with a significant change in ICG-PDR. PMID- 19682846 TI - Study of device use adjusted rates in health care-associated infections after implementation of "bundles" in a closed-model medical intensive care unit. AB - "Bundles" strategies improve health care-associated infection (HCAI) rates in medical intensive care units (MICUs). However, few studies have analyzed HCAI rates adjusted for the device removal component of the bundles. An observational study of adult MICU patients while using bundles to prevent HCAIs associated with endovascular catheters, mechanical ventilation, and urinary tract catheters was conducted. The HCAI rates, unadjusted and adjusted for device use, were calculated using incidence rate ratios (unadjusted IRRs [uIRR] and adjusted IRRs [aIRR], respectively). Among 4550 study patients, HCAIs declined from 47 in 2004 to 10 in 2005, 8 in 2006, and 3 in 2007. Catheter-related blood stream infection (CRBSI) rates decreased from 10.77 to 1.67 per 1000 central line days (uIRR, 0.155; 95% confidence interval [CI], 0.13-0.18; P < .0001). Foley-related urinary tract infections (CA-UTI) decreased from 6.23 to 0.63 per 1000 device days (uIRR, 0.1; 95% CI, 0.08-0.19; P < .0001). Ventilator-associated pneumonia (VAP) per 1000 ventilator days diminished from 2.17 to 0.62 (uIRR, 0.29; 95% CI, 0.21-0.38; P < .0001). After adjustment for device use, aIRRs of CRBSI (0.14; 95% CI, 0.11 0.18), UTI (0.09; 95% CI, 0.06-0.12), and VAP (0.33; 95% CI, 0.22-0.47) declined significantly (P < .00001). Implementing comprehensive bundle strategies reduces HCAI beyond the impact of device removal. PMID- 19682847 TI - Limiting life-sustaining treatment in German intensive care units: a multiprofessional survey. AB - PURPOSE: Deciding about the limitation of life-sustaining treatment (LST) is a major challenge for intensive care medicine. The aim of the study was to investigate the practices and perspectives of German intensive care nurses and physicians on limiting LST. METHODS: We conducted an anonymous, self-administered questionnaire survey among the 268 nurses and 95 physicians on all 10 intensive care units of the Munich University Hospital, Germany. RESULTS: The response rate was 53%. Of all respondents, 91% reported being confronted with the topic at least once a month. Although all reported limiting cardiopulmonary resuscitation, almost no one reported limiting artificial hydration. Half of nurses and junior physicians felt uncertain about the decision-making process. Junior physicians were most dissatisfied with their training for this task and expressed the highest fear of litigation. Nurses were less satisfied than physicians with the communication process. Both nurses and relatives were not routinely involved in decision making. There is no standardized documentation practice, and many notes are not readily accessible to nurses. CONCLUSIONS: Limiting LST is common in German intensive care units. The major shortcomings are team communication, communication with the patient's family, and documentation of the decision-making process. PMID- 19682848 TI - A model for identifying patients who may not need intensive care unit admission. AB - PURPOSE: This study presents a new model for identifying patients who might be too well to benefit from intensive care unit (ICU) care. PATIENTS AND METHODS: Intensive care unit admissions in 2002 to 2003 were used to develop a model to predict whether patients monitored on day one would receive one or more of 33 subsequent active life-supporting treatments. Accuracy was assessed by testing the model in a subsequent cohort of admissions in 2004 to 2006. We then assessed the frequency of active treatment among monitor patients at a low (<10%) risk for active life-supporting therapy on ICU day 1. RESULTS: Among 28 847 ICU monitor admissions in 2004 to 2006, 3153 patients (11.0%) were predicted to receive active treatment; 3296 (11.5%) actually did. There were 17 720 admissions with a low (<10%) risk for receiving subsequent active life-supporting treatment; 1238 (7.0%) received subsequent active treatment. Hospital mortality (2.5%) and mean ICU stay (1.8 days) suggests that most of these patients did not require ICU care. CONCLUSIONS: The outcome for low-risk monitor patients suggest they may be too well to benefit from intensive care. The frequency of low-risk monitor admissions provides a measure of ICU resource use. Improved resource use and reduced costs might be achieved by strategies to provide care for these patients on floors or intermediate care units. PMID- 19682850 TI - Discordance in interpretation of chest radiographs between pediatric intensivists and a radiologist: impact on patient management. AB - PURPOSE: When radiologists are not available, chest radiographs (CXRs) of pediatric intensive care unit (PICU) patients are commonly interpreted by pediatric intensivists. We prospectively investigated the frequency of errors in CXR interpretation by pediatric intensivists and their impact on patient management. MATERIALS AND METHODS: Chest radiographs of PICU patients were evaluated by 5 pediatric intensivists then by a pediatric radiologist (the "gold standard"). If the interpretation of the radiologist and intensivist differed, an independent intensivist determined whether a management change took place. A pediatric pulmonologist determined how many intensivist interpretations were different from the radiologist's interpretations. RESULTS: Seven hundred twenty eight radiographic findings were identified by the radiologist in 460 CXRs. There were 33 interpretation errors by the intensivists (4.5% of the findings in 7.1% of the CXRs). Only 3/33 error corrections (0.45% of the findings in 0.7% of the CXRs) resulted in change in patient management. CONCLUSIONS: Errors in interpretation of CXRs by pediatric intensivists were common but less than that in other series, probably because of education of the pediatric intensivists through daily rounds with the radiologist. Although interpretation errors that affected patient management were rare, their clinical importance supports the growing practice of 24/7 remote radiograph reading by radiologists. PMID- 19682849 TI - The impact of the initial ventilatory strategy on survival in hematological patients with acute hypoxemic respiratory failure. AB - PURPOSE: The aim of this study was to assess the impact of the 3 types of initial respiratory support (noninvasive positive pressure ventilation vs invasive positive pressure ventilation vs supplemental oxygen only) in hematological patients with acute hypoxemic respiratory failure (ARF). MATERIALS AND METHODS: This study is a retrospective analysis of a cohort of hematological patients admitted to the intensive care unit (ICU) of a tertiary care hospital between January 1, 2002, and June 30, 2006. RESULTS: One hundred thirty-seven hematological patients were admitted at the ICU with ARF (defined as Pao(2)/Fio(2) <200): within the first 24 hours, 24 and 67 patients received noninvasive positive pressure ventilation and invasive positive pressure ventilation, respectively, and 46 received supplemental oxygen only. Intensive care unit mortality in the 3 patient categories was 71%, 63%, and 32%, respectively (P = .001), and in-hospital mortality was 75%, 80%, and 47%, respectively (P = .001). In multivariate regression analysis, increasing cancer specific severity-of-illness score upon admission and more organ failure after 24 hours of ICU admission, but not the type of initial respiratory support, were significantly associated with ICU or in-hospital mortality. CONCLUSIONS: Intensive care unit and in-hospital mortality in our population of hematological patients with hypoxemic ARF was determined by severity of illness and not by the type of initial respiratory support. PMID- 19682851 TI - Vasopressin use is associated with death in acute trauma patients with shock. AB - PURPOSE: Traumatic hemodynamic instability is associated with high mortality if not expeditiously corrected. Hypotension despite adequate volume resuscitation is treated with vasopressors. Although catecholamines are typically the first agent used, arginine vasopressin (AVP) is increasingly been used as an adjuvant agent. Mortality with refractory hypotension and vasopressin use in trauma patients is unknown. MATERIALS AND METHODS: A retrospective cohort analysis of trauma patients requiring vasopressors within 72 hours of admission was performed. Two groups were identified: patients who received AVP (AVP+) and those who did not (AVP-). Primary outcome was mortality. RESULTS: Five hundred thirty nine patients met the criteria with 189 patients receiving AVP. Demographics, Injury Severity Score, minimum hemoglobin, and blood volume resuscitation (packed red blood cell, fresh frozen plasma, and platelets) were similar between groups. Trauma and Injury Severity Score suggested a higher probability of survival in AVP+ (0.88 vs 0.73, P < .001); however, the observed mortality was higher (55% vs 41%, P = .002). The age, Injury Severity Score, initial lactate, and severe head injury adjusted odds ratio of death for AVP+ patients was 1.6 (95% confidence interval, 1.1-2.4; P = .02). CONCLUSIONS: Arginine vasopressin is associated with increased mortality in trauma patients with refractory hypotension. Arginine vasopressin may be a marker of illness or possibly play a causal role in adverse outcomes. Clinicians should reconsider expanding the indications of AVP use. PMID- 19682852 TI - Body mass index is negatively correlated with respiratory muscle weakness and interleukin-6 production after coronary artery bypass grafting. AB - PURPOSE: The present study was performed to clarify the relationships between body mass index (BMI), interleukin-6 (IL-6) production, and respiratory muscle weakness in patients undergoing coronary artery bypass grafting (CABG). MATERIALS AND METHODS: The correlations among BMI, changes in maximum inspiratory and expiratory pressure (DeltaMIP, DeltaMEP) on postoperative days (POD) 1 and 7, postoperative IL-6, and rapid turnover proteins (retinol-binding protein, prealbumin, and transferrin) on POD1 were assessed in 154 consecutive patients undergoing elective CABG. The patients were divided into quartiles of BMI, Q1 (BMI, <20.8 kg/m(2)) to Q4 (BMI, > or =25.25 kg/m(2)), and compared among groups. RESULTS: There were significant correlations between DeltaMIP and BMI (POD1, r = 0.369; POD7, r = -0.285) and IL-6 (POD1, r = 0.423; POD7, r = 0.431), and between DeltaMEP and BMI (POD1, r = -0.252; POD7, r = -0.228) and IL-6 (POD1, r = 0.252; POD7, r = 0.384). Interleukin-6 showed a significant negative correlation with BMI (r = -0.374) and retinol-binding protein (r = -0.382). Interleukin-6 was highest in Q1 and lowest in Q4. Higher BMI indicated greater respiratory muscle strength than lower BMI. CONCLUSIONS: Preoperative BMI is correlated with respiratory muscle strength and cytokine production after CABG. The findings of this study suggest that BMI may be a valuable predictor for respiratory management in CABG patients. PMID- 19682853 TI - Can an adequate energy intake be able to reverse the negative nitrogen balance in mechanically ventilated critically ill patients? AB - PURPOSE: Adequate energy provision and nitrogen losses prevention of critically ill patients are essentials for treatment and recovery. The aims of this study were to evaluate energy expenditure (EE) and nitrogen balance (NB) of critically ill patients, to classify adequacy of energy intake (EI), and to verify adequacy of EI capacity to reverse the negative NB. METHODS: Seventeen patients from an intensive care unit were evaluated within a 24-hour period. Indirect calorimetry was performed to calculate patient's EE and Kjeldhal for urinary nitrogen analysis. The total EI and protein intake were calculated from the standard parenteral and enteral nutrition infused. Underfeeding was characterized as EI 90% or less and overfeeding as 110% or greater of EE. The adequacy of the EI (EI EE(-1) * 100) and the NB were estimated and associated with each other by Spearman coefficient. RESULTS: The mean EE was 1515 +/- 268 kcal d(-1), and most of the patients (11/14) presented a negative NB (-8.2 +/- 4.7 g.d(-1)). A high rate (53%) of inadequate energy intake was found, and a positive correlation between EI EE(-1) and NB was observed (r = 0.670; P = .007). CONCLUSION: The results show a high rate of inadequate EI and negative NB, and equilibrium between EI and EE may improve NB. Indirect calorimetry can be used to adjust the energy requirements in the critically ill patients. PMID- 19682854 TI - Temporal allocation of attention toward threat in individuals with posttraumatic stress symptoms. AB - Research suggests that individuals with posttraumatic stress disorder (PTSD) selectively attend to threat-relevant information. However, little is known about how initial detection of threat influences the processing of subsequently encountered stimuli. To address this issue, we used a rapid serial visual presentation paradigm (RSVP; Raymond, J. E., Shapiro, K. L., & Arnell, K. M. (1992). Temporary suppression of visual processing in an RSVP task: An attentional blink? Journal of Experimental Psychology: Human Perception and Performance, 18, 849-860) to examine temporal allocation of attention to threat related and neutral stimuli in individuals with PTSD symptoms (PTS), traumatized individuals without PTSD symptoms (TC), and non-anxious controls (NAC). Participants were asked to identify one or two targets in an RSVP stream. Typically processing of the first target decreases accuracy of identifying the second target as a function of the temporal lag between targets. Results revealed that the PTS group was significantly more accurate in detecting a neutral target when it was presented 300 or 500ms after threat-related stimuli compared to when the target followed neutral stimuli. These results suggest that individuals with PTSD may process trauma-relevant information more rapidly and efficiently than benign information. PMID- 19682856 TI - Limited validity of parental recall on pregnancy, birth, and early childhood at child age 10 years. AB - OBJECTIVE: Evidence on the validity of parental recall of early childhood behavior is lacking. Our aim was to examine the validity of parental recall at child age 10-12 years for maternal lifestyle during pregnancy, the birth characteristics, and early childhood behavior. STUDY DESIGN AND SETTING: The study population comprised 2,230 children and their parents. Children aged 10-12 years were recruited from elementary schools (response: 76.0%). Parents were asked to recall lifestyle during pregnancy, birth characteristics, and childhood behavior at age 4-6 years. Recalled data were compared with information registered by Preventive Child Healthcare (PCH) from birth onwards. RESULTS: For birth weight and gestational age, we found no systematic difference between recalled and PCH-registered data; 95% limits of agreement were + or - 1.2 pounds (600 g) and + or - 2.4 weeks, respectively. For maternal alcohol use during pregnancy and early childhood behavior problems, Cohen's kappas were low (0.03 0.11). Compared with PCH registration, parents tended to overreport at age 10-12 years. In contrast, kappa was high for maternal smoking during pregnancy (0.77). CONCLUSION: Retrospectively collected information on lifestyle during pregnancy, birth, and early childhood behavior is sometimes biased, which limits its value in estimating the contribution of early-life adversity to health in later life. PMID- 19682855 TI - The Short-Form Six-Dimension utility index predicted mortality in the European Prospective Investigation into Cancer-Norfolk prospective population-based study. AB - OBJECTIVE: To examine the relationship between the Short-Form Six-Dimension (SF 6D) and mortality. STUDY DESIGN AND SETTING: Participants were 17,736 men and women aged 40-79 years at baseline who lived in Norfolk, UK, and had no known cardiovascular disease or cancer, and completed the anglicized Short-Form 36 (SF 36)-item during 1996-2000 in the European Prospective Investigation into Cancer Norfolk prospective population study. The SF-36 data were converted to SF-6D. The relationship between SF-6D and all-cause and cause-specific mortality were examined. RESULTS: One thousand and seventy deaths occurred during a total of 115,255 person years of follow-up (mean 6.5 years). Lower SF-6D was associated with increased risk of all-cause mortality in men and women. A decrease of 1 standard deviation (0.12 point) in SF-6D was associated with a 35% increase in all-cause mortality (hazards ratio = 1.35; 95% CI: 1.26, 1.45) after controlling for age, gender, body mass index, systolic blood pressure, cholesterol, diabetes, smoking, and social class. Similar results were observed for cardiovascular, cancer, and other causes of deaths. CONCLUSION: Poor health utility measured by the SF-6D predicted increased risk of all-cause and cause-specific mortality in men and women. The present study provides the first evidence of the sensitivity of the SF-6D in predicting mortality in an apparently healthy population. PMID- 19682857 TI - Navigating the passage between Charybdis and Scylla: recognizing the achievements of Noel Rose. AB - This special edition of the Journal of Autoimmunity is dedicated to celebrate the enormous contributions of Dr. Noel Rose, a physician-scientist, someone that can be readily anointed as a gifted scientist who laid down the foundation and fundamental concepts of human autoimmunity. Dr. Rose performed a series of landmark studies that extend back more than 50 years and whose work is the cornerstone of the commonly used term "from the bench to the bedside." The studies included not only characterization of a normal immune response but, more importantly, defining the nature of not only the antigen, but also the aberrant response that results in organ, tissue specific reactions and immunopathology. These pioneering studies contributed to the biochemical nature of antigens, dissection of the immune repertoire, the recognition of the importance of genetics and environment, and the distinctions between a normal and an autoimmune response. Hence, this volume has been coined "Navigating the Passage Between Charybdis and Scylla: Recognizing the Achievements of Noel Rose." PMID- 19682858 TI - The genetics of human autoimmune disease. AB - Autoimmune diseases are known to have a multifactorial pathogenesis, with both environmental and inherited components. Wide technical progresses together with the completion of the sequencing of human genome have recently allowed the identification of new genetic risk variants in many autoimmune disorders. While part of these studies confirmed previous knowledge, most of the data has disclosed novel and unsuspected roles in the development of autoimmunity for molecules involved in various pathogenic pathways. After the current first wave of data from high-density genome-wide studies, we now need to further characterize these genetic factors and find additional ones, possibly among rare variants. In addition, a role for sex chromosomes in the development of autoimmune diseases has also been suggested. This review will focus on the recent discoveries related to genetics of autoimmunity. PMID- 19682860 TI - Long-term results of surgical therapy for masticatory muscle tendon-aponeurosis hyperplasia accompanied by limited mouth opening. AB - Masticatory muscle tendon-aponeurosis hyperplasia is a new disease entity characterized by limited mouth opening due to contracture of the masticatory muscles, resulting from hyperplasia of tendons and aponeuroses. In the case of masseter muscle type, the face displays a square mandible configuration. Pharmacotherapy, occlusal splints and physical therapy are ineffective. This study evaluated the long-term results of aponeurectomy of the masseter muscle with coronoidectomy to release the temporal muscle tendon. The subjects were 10 patients who underwent surgery between 2000 and 2005. Mean maximum mouth opening before surgery was 21.8mm (range 17-29 mm). All patients received bilateral aponeurectomy of the masseter muscle and coronoidectomy. Three patients additionally underwent bilateral anglectomy for esthetic reasons. After discharge, one patient did not return to the hospital. Data from the other nine patients were analyzed. The mean duration of follow-up was 4 years. At final follow-up, the maximum mouth opening was >44 mm in four patients, 40-44 mm in three patients, and 35-39 mm in two patients. Overall satisfaction was excellent or good in all patients. PMID- 19682859 TI - Application of tissue-specific NK and NKT cell activity for tumor immunotherapy. AB - Natural killer (NK) and NKT cells are a first line of defense against pathogens and transformed cells. However, dysregulation of their function can lead to autoimmune disease. A better understanding of the mechanisms controlling NK and NKT effector function should lead to the development of improved strategies for the treatment of many diseases. The site in which NK and NKT cells reside should be taken into account, because accumulating evidence suggests that the tissue microenvironment strongly influences their function. In this regard, the liver represents a unique immunologic organ in which the balance between the need for tolerance and the ability to respond rapidly to pathogens and tissue injury is tightly regulated. NK cells in the liver have augmented cytolytic activity as compared to other organs, which is consistent with a role for liver-associated NK cells in being critical effector cells for inhibiting tumor metastasis in the liver. Several studies also suggest that hepatic NKT cells have different functions than those in other organs. Whereas splenic and thymic NKT cells have been shown to suppress diabetes development, facilitate the induction of systemic tolerance and are regulated by IL-4 and other Th2 cytokines, certain subsets of NKT cells in the liver are important sources of Th1 cytokines such as Interferon gamma, and are the primary mediators of anti-tumor responses. The unique properties and roles as critical effector cells make NK and NKT cells within the liver microenvironment attractive targets of immunotherapeutic approaches that have the goal of controlling tumor metastasis in the liver. PMID- 19682861 TI - Association study of the gamma-aminobutyric acid type a receptor gamma2 subunit gene with schizophrenia. AB - Schizophrenia (SCZ) is a severe neuropsychiatric disorder with a strong genetic basis. We analyzed eight GABRG2 and one DRD5 tag single-nucleotide polymorphisms for association with SCZ in 109 small nuclear families and 229 independent SCZ case-control pairs. The marker rs183294 in the 5' region of GABRG2 was found to be associated with SCZ in both samples with the C allele over-represented in SCZ cases and over-transmitted in SCZ families (combined z=9.18; p<1 x 10(-3)). Taken together, the results of the present study suggest that GABRG2 may be involved in SCZ susceptibility, but further studies are required. PMID- 19682862 TI - Efficacy of moxifloxacin or gatifloxacin as prophylaxis against experimental murine Brucella melitensis infection. AB - The prophylactic potential of moxifloxacin and gatifloxacin was assessed in comparison with doxycycline, an established therapeutic antibiotic, to limit or control infection by Brucella melitensis in an experimental mouse model, determined by reduced bacterial burden in the spleen. Although moxifloxacin was found to have a small protective effect when administered 6 h following infection, neither moxifloxacin nor gatifloxacin showed significant efficacy in vivo. In comparison, doxycycline provided significant protection when prophylaxis was started at 6 h, 7 days or 14 days following infection. Overall, these results confirm the utility of doxycycline in the prophylaxis of brucellosis and suggest that neither moxifloxacin nor gatifloxacin are likely to be valuable for post exposure prophylaxis of Brucella infection. PMID- 19682863 TI - Cardiovascular disease and diabetes in people with severe mental illness position statement from the European Psychiatric Association (EPA), supported by the European Association for the Study of Diabetes (EASD) and the European Society of Cardiology (ESC). AB - People with severe mental illnesses, such as schizophrenia, depression or bipolar disorder, have worse physical health and reduced life expectancy compared to the general population. The excess cardiovascular mortality associated with schizophrenia and bipolar disorder is attributed in part to an increased risk of the modifiable coronary heart disease risk factors; obesity, smoking, diabetes, hypertension and dyslipidaemia. Antipsychotic medication and possibly other psychotropic medication like antidepressants can induce weight gain or worsen other metabolic cardiovascular risk factors. Patients may have limited access to general healthcare with less opportunity for cardiovascular risk screening and prevention than would be expected in a non-psychiatric population. The European Psychiatric Association (EPA), supported by the European Association for the Study of Diabetes (EASD) and the European Society of Cardiology (ESC) published this statement with the aim of improving the care of patients suffering from severe mental illness. The intention is to initiate cooperation and shared care between the different healthcare professionals and to increase the awareness of psychiatrists and primary care physicians caring for patients with severe mental illness to screen and treat cardiovascular risk factors and diabetes. PMID- 19682864 TI - Spectroscopic correlates of antidepressant response to sleep deprivation and light therapy: a 3.0 Tesla study of bipolar depression. AB - Glutamate is the primary excitatory neurotransmitter of the human brain, and recent findings suggest a role for the glutamatergic system in the pathophysiology and treatment of mood disorders. Single proton magnetic resonance spectroscopy (1H-MRS) was used to study the relative in vivo levels of brain neural metabolites. We evaluated the effect of antidepressant treatments on the relative concentration of unresolved glutamate and glutamine (Glx) with GABA contamination (2.35 ppm peak) using single voxel 1H-MRS at 3.0 Tesla. We studied 19 inpatients (7 males, 12 females) affected by bipolar disorder type I, current depressive episode without psychotic features, before and after 1 week of treatment with repeated total sleep deprivation (TSD) combined with light therapy (LT). Chronobiological treatment caused a significant amelioration in mood levels. Changes in the brain Glx/creatine ratio followed a general trend toward decrease, with individual variability. We observed that the decrease in the Glx/creatine ratio significantly correlated with the improvement of both objective and subjective measures of depression. PMID- 19682865 TI - Regional distribution and behavioral correlates of 5-HT(2A) receptors in Alzheimer's disease with [(18)F]deuteroaltanserin and PET. AB - Postmortem studies show reductions in brain serotonin 2A (5-HT(2A)) receptors in Alzheimer's disease (AD). Converging evidence also suggests that serotonergic dysregulation may contribute to behavioral symptoms that frequently occur in AD. This study aimed to define regional reductions in 5-HT(2A) binding in AD patients and to examine their behavioral correlates. Nine patients with probable AD and eight elderly controls were studied using a constant infusion paradigm for equilibrium modeling of [(18)F]deuteroaltanserin with positron emission tomography (PET). Region of interest analyses were performed on PET images coregistered to MRI scans. The outcome measures BP(P) (ratio of specific brain uptake to total plasma parent concentration) and BP(ND) (ratio of specific to nondisplaceable uptake) were obtained for pertinent cortical and subcortical regions. AD patients showed a statistically significant decrease in the anterior cingulate in both BP(P) and BP(ND), but in no other region. Within the AD patient sample, no significant correlations were observed between regional 5-HT(2A) binding and behavioral measures, including depressive and psychotic symptoms. These results confirm a reduction in cortical 5-HT(2A) receptors in AD, specifically in the anterior cingulate. However, in a limited AD patient sample, they fail to demonstrate a relationship between regional 5-HT(2A) binding and major behavioral symptoms. PMID- 19682866 TI - Depressive mood in pre-dialytic chronic kidney disease: Statistical parametric mapping analysis of Tc-99m ECD brain SPECT. AB - The purpose of this study was to investigate depression-related regions in pre dialytic patients with chronic kidney disease (CKD) patients. Participants comprised 33 patients with stage 4 and 5 CKD (age, 55 [42-63]) and 32 healthy volunteers (age, 53.5 [49.5-57]). Depressed mood was assessed in the patients, and both groups underwent Tc-99m-labeled ethylcysteinate dimer (Tc-99m ECD) single photon emission computed tomograpy (SPECT). Statistical parametric mapping identified 18 areas of hypoperfusion in the patients in comparison with the normal controls. The largest clusters were areas including left precentral gyrus, right superior and middle temporal gyrus, both cerebellar posterior lobes, both inferior frontal gyrus, right superior and middle frontal gyrus, right cuneus, right inferior parietal lobule, and right putamen. However, there were no specific hypoperfusion areas in CKD patients with depression compared with CKD patients without depression. Interestingly, several hypoperfusion areas in CKD patients (inferior frontal gyrus [BA46], superior temporal gyrus [BA42], anterior cingulate gyrus [BA24]) were concordant with hypoperfusion areas found in patients with major depression who were free of kidney disease. In conclusion, this study did not demonstrate specific depression-related cerebral hypoperfusion areas. However, the cerebral blood flow pattern in CKD patients was similar to that of patients with major depression in some areas. Although further investigations are needed in the future, we suggest that the causes of the higher prevalence of depression in CKD might be associated with this finding. PMID- 19682867 TI - Gender difference in relationship between anxiety-related personality traits and cerebral brain glucose metabolism. AB - Recent functional neuroimaging studies have suggested that specific brain regions might be associated with the formation of anxiety-related personality traits, which are well known to be influenced by gender. Such anxiety-related personality traits are one of the representative predisposing factors for mood and anxiety disorders, whose incidence is also known to be much influenced by gender. However, little is known about the gender differences in brain function related to anxiety-related personality traits. The aim of the present study was to examine gender-related differences in the pattern of the relationships between an anxiety-related personality trait and cerebral brain glucose metabolism. Regional brain glucose metabolism was measured using [(18)F]fluorodeoxyglucose positron emission tomography in 102 healthy subjects (65 males and 37 females). An anxiety related trait was assessed using the Temperament and Character Inventory dimension Harm Avoidance (HA). HA was negatively correlated with glucose metabolism in the anterior portion of the ventromedial prefrontal cortex (vmPFC) in females but not in males. The anterior vmPFC may be a possible neural target for the prevention or therapy of emotional disorders, especially in females. PMID- 19682868 TI - Combined solid state NMR and ONIOM studies of reversible crystalline phase reaction for nickel coordination compounds. AB - Ni(II) with bis(acetylacetone)ethylenediamine ligand forms complexes which crystallizes as semi-hydrate with C2/c space group in monoclinic system and anhydrous form with Pna2(1) space group in orthorhombic system. (13)C and (15)N CP/MAS experiments were employed for structural characterization of both forms and searching of process of reversible water exchange in the crystal lattice. Comparative analysis of (13)C and (15)N principal elements of chemical shift tensors for ligand and complexes showed the sensitivity of delta(ii) elements both for complexation and phase reaction processes. Theoretical (13)C and (15)N NMR shielding parameters sigma(ii) were computed employing ONIOM approach and correlated with experimental delta(ii) data. The applicability of ONIOM in structural analysis of coordination compounds (CC) is discussed. PMID- 19682869 TI - Soft matter dispersions with ordered inner structures, stabilized by ethoxylated phytosterols. AB - This paper describes the formation and characterization of liquid crystalline dispersions based on the hexagonal phase of GMO/tricaprylin/water. As a stabilizer of the soft particles dispersed in the aqueous phase, a non-ionic, non polymeric surfactant--ethoxylated phytosterol with 30 oxyethylene units (PhEO) was utilized. In contrast to Pluronic copolymers, normally utilized in the stabilization of liquid crystalline dispersions with ordered inner structure, use of such non-polymeric surfactant is not a common practice in this field. We revealed how properties of these particles, such as internal structure, size, and stability, can be rationally modified by the concentration of the stabilizing agent and processing conditions. The physical stability of the hexosomes was further examined by the LUMiFuge technique. Structural effect of PhEO solubilization on the properties of the bulk H(II) mesophase system showed that phase behavior was greatly influenced following phase transitions: H(II)- >H(II)+cubic-->cubic+L(alpha)-->L(alpha). The decrease of hydrogen bonding of the hydroxyl and carbonyl groups of monoolein with water and simultaneous hydration of EO groups of PhEO appeared to be important for the observed behavior. The use of PhEO as a dispersant resulted in a soft matter multi-phase water dispersion with bimodal distribution of the particle population. Effective stabilization of hexosomes was obtained in an extremely narrow concentration range of PhEO (0.1 0.2 wt%), coexisting with small vesicles and disordered particles. At higher PhEO content, particles had disordered inner structure, and unilamellar and multilamellar vesicles, at the expense of hexosomes in consequence of incorporation of the dispersant into the hexosome structure. PhEO was found to induce lamellar phase formation, introducing disorder into the hexagonal LLC and reducing their domain size. Finally, hexosomes were evaluated as delivery vehicles for the therapeutic peptide desmopressin. Sustained release of this drug was observed during the first 10 h; however, permeation drastically increased in the 10-24 h range. PMID- 19682870 TI - Surface active properties of chitosan and its derivatives. AB - This review discusses the definition of surface active agents and specifically natural polymeric surface active agents. Chitosan by itself was found to have weak surface activity since it has no hydrophobic segments. Chemical modifications of chitosan could improve such surface activity. This is achieved by introducing hydrophobic substituents in its glucosidic group. Several examples of chitosan derivatives with surfactant activity have been surveyed. The surface active polymers form micelles and aggregates which have enormous importance in the entrapment of water-insoluble drugs and consequently applications in the controlled drug delivery and many biomedical fields. Chitosan also interacts with several substrates by electrostatic and hydrophobic interactions with considerable biomedical applications. PMID- 19682871 TI - Orientation and distance dependent chiral discrimination in the first step of the aminoacylation reaction: integrated molecular orbital and semi-empirical method (ONIOM) based calculation. AB - Aminoacylation is a vital step in natural biosynthesis process of peptide and is the key step in correlating the realm of protein with the RNA world. Incorrect aminoacylation might lead to misacylation of d-amino acid in the tRNA which might cause synthesis of a hetero-peptide rather than natural homopeptide leading to the altered functionality of the peptide. However, the accuracy of this process is remarkable and leads to the attachment of the correct enantiomer of the amino acid with their cognate tRNA. Thus, the chiral discrimination is stringent. In the present work, we presented a combined ONIOM (ab initio/semi-empirical) study of the chiral discrimination in the first step of aminoacylation reaction based on a model of crystal structure of the oligomeric complex of histidyl-tRNA synthetase (HisRS) from Escherichia coli complexed with ATP and histidinol and histidyl-adenylate. The study reveals that the molecular mechanism of the chiral discrimination involves the amino acid, ATP as well as surrounding residues of the synthetase. Several factors are noted to be responsible for discrimination and explain the high level of stereospecificity of the process. The chirality of the amino acid of the substrate and its (principally) electrostatic interaction with the ATP is important for discrimination. The distance and orientational changes involved in the approach of the d-His towards the ATP is energetically unfavorable. The charge distributions on the His and ATP are important for the discrimination. Removal of the charges in the model drastically reduces the discrimination. Restricted nature of the mutual orientation within the cavity of the active site where the His and ATP are located during the change in orientation for the approach to form the adenylate makes the resultant interaction profile as different for l-His and d-His also influences chiral discrimination. The analysis of the transition state structure revealed that alteration of the chirality of the His destabilize the transition state by removing the favorable electrostatic interaction between the Glu-83 and NH(3)(+) group of the His substrate. The proximity of the surrounding residues as present in the active site of the synthetase with the His and ATP (the separation is of nanometer range) has influence of discrimination. The study provides a molecular mechanism of the retention of biological homochirality. PMID- 19682872 TI - Hypoglycemic and hypolipidemic effect and antioxidant activity of chronic epigallocatechin-gallate in streptozotocin-diabetic rats. AB - BACKGROUND AND OBJECTIVE: Green tea catechins including epigallocatechin-3 gallate (EGCG) could exert beneficial health effects to ameliorate metabolic diseases. The effect of chronic administration of EGCG was evaluated on serum glucose and lipid profile and hepatic lipid peroxidation in streptozotocin (STZ) diabetic rats. MATERIALS AND METHODS: Rats received EGCG 25mg/kg/day for 8 weeks 1 week after diabetes induction. Serum glucose, triglyceride, total cholesterol, HDL- and LDL-cholesterol levels and MDA level and SOD activity in hepatic tissue were spectrophotometrically measured. RESULTS: Treatment of diabetic rats with EGCG produced a hypoglycemic effect and there were appropriate changes regarding serum lipids in treated diabetic group. Meanwhile, EGCG treatment attenuated the increased MDA content and reduced activity of SOD in liver. CONCLUSION: Chronic treatment of diabetic rats with EGCG could prevent abnormal changes in blood glucose and lipid profile and attenuat hepatic lipid peroxidation. PMID- 19682873 TI - Pathophysiologic classification of stroke territory (PCST). AB - Pathophysiologic Classification of Stroke Territory (PCST) was designed for the categorization of brain infarctions. It is based on the neuroimaging and etiology of stroke. PCST provides an impression about mechanism of stroke in clinical practice. PMID- 19682874 TI - Growth and gaseous emissions of pure and mixed small intestinal bacterial cultures: Effects of bile and vancomycin. AB - Simultaneous cultivations in anaerobiosis, aerobiosis and with microaerobic gas mixture were used to clarify the bile (oxgall) effects on the pure and mixed cultures of enterobacterial strains in simulations in Portable Microbe Enrichment Unit (PMEU) linked with ChemPro100i((R)) gas detector. The effects of vancomycin were evaluated in aerobic cultures. Growth and metabolic activity of cultures were also followed by measuring sugar consumption, pH alterations, and colony counts on BD CHROMagar Orientation plates. Results showed that the two fermentatively different strains of facultative anaerobes, Escherichia coli E 17 and Klebsiella mobilis ATCC 13048 grew in balance regardless of oxygen level, bile acid concentration or other components of the mixed cultures, Bacillus cereus or Staphylococcus aureus. When the evaporations of the mixed cultures of E. coli, K. mobilis and S. aureus were compared with the emissions of the corresponding pure cultures by ChemPro100i((R)) gas sensing detector, the pure cultures of bile resistant E. coli and K. mobilis produced more gaseous components than the mixed culture indicating that these organisms cooperate and use the substrate more effectively together than separately. A survey of the aseptic bacterial isolations from the bile tract in a big University Hospital, (Salzburg, Austria) during 3 years, showed that these bacterial groups dominated. Only 13.24% of the 287 patient samples were sterile, and around 180 strains of both E. coli and Klebsiella/Enterobacter groups were found amongst 973 isolates from 249 patients (together 35.57%). Enterococcus sp. accounted for 246 isolates being the largest group of strains (24.25% of all the isolates). In anaerobiosis it was shown that Klebsiella neutralized the acids produced in the mixed acid fermentation of the E. coli. The ethanol produced from both groups evaporated in the gas stream of the PMEU culturing step and its formation also removes excess acidity from the cultures. The synergistic behaviour and symbiotic function between E. coli and Klebsiella/Enterobacter strains is suggested. PMID- 19682875 TI - Application of robotic stereotactic radiotherapy to peripheral stage I non-small cell lung cancer with curative intent. AB - AIMS: To determine the effectiveness of robotic stereotactic radiotherapy with image guidance and real-time respiratory tracking against early stage peripheral lung cancer. MATERIALS AND METHODS: We treated patients with stage I non-small cell lung cancer (NSCLC) with CyberKnife and analysed their clinical characteristics and outcomes. All patients had co-morbid conditions that precluded lobectomy. The clinical target volume (CTV) included the gross tumour volume (GTV) and a 6mm margin in all directions to account for microscopic extension. The planning target volume (PTV) equalled CTV+2mm in all directions for uncertainty. Tumour motion was tracked using a combination of Synchrony and Xsight Spine tracking methods with the aid of a single gold marker implanted in the centre of the tumour, or using the newer Xsight Lung method without markers for selected tumours. A 60-67.5 Gy dose was prescribed to the 60-80% isodose line (median 65%) and given in three to five fractions. Patients were followed every 3 months for a median of 27.5 months (range 24-53 months). RESULTS: Of the 67 patients with NSCLC stage IA or IB treated between January 2004 and December 2008, we report the results of a cohort of 31 with peripheral stage I tumours of 0.6-71 cm(3) volume treated between January 2004 and December 2007 with total doses between 60 and 67.5 Gy in three to five fractions. The median D(max) was 88.2 Gy and the median V(95) of the PTV was 99.6% or 27.9 cm(3). No grade 3 or above toxicity was encountered. Four cases of radiation pneumonitis and one case of oesophagitis were observed. In those patients whose pre- and post-treatment results were available, no change in pulmonary function tests was observed. Actuarial local control was 93.2% for 1 year and 85.8% for up to 4.5 years. One year overall survival was 93.6% and 83.5% for up to 4.5 years, as projected by Kaplan-Meier analyses. CONCLUSIONS: In this small cohort of patients with stage I peripheral NSCLC, robotic stereotactic radiotherapy seems to be a safe and obviously superior alternative to conventionally fractionated radiotherapy, with results that may be approaching those obtained with lobectomy without the associated morbidity. PMID- 19682876 TI - Mechanism of herpes simplex virus type 2 suppression by propolis extracts. AB - Genital herpes caused by herpes simplex virus type 2 (HSV-2) is a chronic, persistent infection spreading efficiently and silently as sexually transmitted disease through the population. Antiviral agents currently applied for the treatment of herpesvirus infections include acyclovir and derivatives. Aqueous and ethanolic extracts of propolis were phytochemically analysed, different polyphenols, flavonoids and phenylcarboxylic acids were identified as major constituents. The aqueous propolis extract revealed a relatively high amount of phenylcarboxylic acids and low concentrations flavonoids when compared to the ethanolic special extract GH 2002. The cytotoxic and antiherpetic effect of propolis extracts against HSV-2 was analysed in cell culture, and revealed a moderate cytotoxicity on RC-37 cells. The 50% inhibitory concentration (IC(50)) of aqueous and ethanolic GH 2002 propolis extracts for HSV-2 plaque formation was determined at 0.0005% and 0.0004%, respectively. Both propolis extracts exhibited high levels of antiviral activity against HSV-2 in viral suspension tests, infectivity was significantly reduced by >99% and a direct concentration- and time-dependent antiherpetic activity could be demonstrated for both extracts. In order to determine the mode of virus suppression by propolis, the extracts were added at different times during the viral infection cycle. Addition of these drugs to uninfected cells prior to infection or to herpesvirus-infected cells during intracellular replication had no effect on virus multiplication. However both propolis extracts exhibited high anti-herpetic activity when viruses were pretreated with these drugs prior to infection. Selectivity indices were determined at 80 and 42.5 for the aqueous and ethanolic extract, respectively, thus propolis extracts might be suitable for topical therapy in recurrent herpetic infection. PMID- 19682877 TI - Pharmacological studies on the sedative-hypnotic effect of Semen Ziziphi spinosae (Suanzaoren) and Radix et Rhizoma Salviae miltiorrhizae (Danshen) extracts and the synergistic effect of their combinations. AB - Semen Ziziphi spinosae (Suanzaoren in China) and Radix et Rhizoma Salviae miltiorrhizae (Danshen in China) are conventional herbal drugs in traditional Chinese medicine and have been used widely for the treatment of insomnia. In the present study, the sedative-hypnotic activity of the active fractions extracted from Suanzaoren and Danshen were studied using the method of pentobarbital induced sleep in the mouse model. Qualitative analysis of the standardized extracts was carried out by HPLC-DAD. The results showed that the water extract of Suanzaoren (SWE) (400 and 800 mg/kg body wt.) and the ether extract of Danshen (DTT) (300 and 600 mg/kg body wt.) can shorten sleep latency significantly, increase sleeping time and prolong movement convalescence time induced by sodium pentobarbital (55 mg/kg body wt.) administration in mice. Furthermore, the combination of SWE and DTT showed significant synergistic effect (p<0.05) in decreasing sleep latency and increasing sleeping time, but not in prolonging the movement convalescence time, which might be helpful for energy recovery in the treatment of insomnia. The results suggest that SWE, DTT, and the combination of SWE and DTT possess significant sedative-hypnotic activity, which supports the popular use of Suanzaoren and Danshen for treatment of insomnia and provide the basis for new drug discovery. Furthermore, the results demonstrate that the combination of SWE and DTT may be preferable for the treatment of insomnia. PMID- 19682879 TI - Antitumor activity of terpenoids against classical and atypical multidrug resistant cancer cells. AB - Nineteen terpenoids, including macrocyclic diterpenes, diterpenic lactones and other polycyclic diterpenes, steroids and a triterpene isolated from the methanolic extracts of Euphorbia species, were evaluated for their potential antineoplastic activity in various human cancer cell lines that were derived from three tumor entities: gastric (EPG85-257), pancreatic (EPP85-181) and colon (HT 29) carcinomas. Furthermore, different multidrug-resistant variants of these cancer cell lines with over-expression of MDR1/P-gp or no MDR1/P-gp expression were also investigated. In parental drug-sensitive cell lines, the tested compounds showed a moderate/weak antiproliferative effect or were inactive. Most of them were found more effective in drug-resistant cells than in the parental, drug-sensitive ones, and some of them showed high antineoplastic efficacy in classical or atypical drug-resistant cells. The most active compounds were the lathyrane diterpenes latilagascenes C and D, and the diterpenic lactones 3beta acetoxy-helioscopinolide B and helioscopinolide E which exhibited high antineoplastic activities against the drug-resistant subline EPG85-257RDB derived from gastric carcinoma. In addition, the macrocyclic lathyrane diterpene jolkinol B was found to be highly effective in the multidrug-resistant variant HT-29RNOV. PMID- 19682878 TI - The administration of demethyl fruticulin A from Salvia corrugata to mammalian cells lines induces "anoikis", a special form of apoptosis. AB - Recently demethyl fruticulin A was identified as the major diterpenoid component of the exudates produced by the trichomes of Salvia corrugata leafs. Given the documented apoptotic effects of some of the other known components of the exudates from Salvia species, we assessed if demethyl fruticulin A, once administered to mammalian cells, was involved in the onset of apoptosis and if its biological effects were exerted through the participation of a scavenger membrane receptor, CD36. Three model cell lines were chosen, one of which lacking CD36 expression. Functional availability of the receptor, or its transcriptional rate, were blocked/reduced with a specific antibody or by the administration of vitamin E. Immunodetection of cell cytoskeletal components and tunel analysis revealed that demethyl fruticulin A triggers the onset of anoikis, a special form of apoptosis induced by cell detachment from the substrate. Impairment of CD36 availability/transcription confirmed the receptor partial involvement in the intake of the substance and in anoikis, as also sustained by FACS analysis and by the downregulation of p95, a marker of anoikis, upon blockade of CD36 transcription. However, experiments with CD36-deficient cells suggested that alternate pathways, still to be determined, may take part in the biological effects exerted by demethyl fruticulin A. PMID- 19682880 TI - Escin improves sperm quality in male patients with varicocele-associated infertility. AB - PURPOSE: To evaluate the efficacy of escin to improve sperm quality in Chinese male patients with varicocele-associated infertility. METHODS: This study included 219 male patients with varicocele-associated infertility. These patients were randomly assigned into three groups: the control, the surgery and the escin group. 106 patients, with a mean age of 30 years old and a mean period of infertility of 5.0 years, were recruited in the escin group and a daily dose of 60mg (30mg every 12h) of escin was administered orally during an uninterrupted period of 2 months. The diameter of spermatic vein was also measured using color Doppler ultrasonography before and after treatment in the escin group. Patients in all groups took composite medicines favorable for sperm quality and semen were obtained and analyzed before and after treatment. RESULTS: In response to treatment, the improvement rates in sperm density in the control, the surgery and the escin group, were 38.5%, 68.8%, and 57.5%, respectively. The differences in the surgery and the escin groups were significant compared to that in the control group (68.8% or 57.5% vs. 38.5%, P<0.05). The improvement rates in sperm motility in the control, the surgery and the escin group were 46.2%, 77.1%, and 55.7%, respectively. Significant difference was only observed between the surgery and the control group (77.1% vs. 46.2%, P<0.05). In the escin group, when severity of varicocele was classified to mild, moderate or severe degree according to the diameter of the spermatic vein, the improvement rates in disease severity were higher in the mild (41.7% vs. 20.0%, P<0.05) and moderate severity subgroups (64.4% vs. 20.0%, P<0.05) when compared to that in the severe subgroup (20.0%). The improvement rate in disease severity was also higher in the moderate subgroup when compared to that in the mild subgroup (64.4% vs. 41.7%, P<0.05). During the period of treatment, only very low frequencies of mild adverse effects were observed, most of which resolved without further symptomatic drug therapy after advising the patients to take escin after meal. Escin has little effect on vital signs, blood counts, liver or kidney function. CONCLUSION: Escin is a safe and effective drug to improve sperm quality in Chinese male patients with varicocele associated infertility. PMID- 19682881 TI - Anti-stress effects of the "tonic"Ptychopetalum olacoides (Marapuama) in mice. AB - With the recognition that high levels of sustained stress are associated with the natural course of countless illnesses, effective anti-stress agents have gained importance. Improved endurance to particularly stressful periods is one of the medicinal claims for Marapuama (Ptychopetalum olacoides Bentham, PO), a popular Amazonian herbal. The purpose of this study was to evaluate if PO possesses anti stress properties. To this end, an extract from PO (POEE) was evaluated on anxiety and glucose levels in mice submitted to the unpredictable chronic mild stress (UCMS) paradigm. POEE did not present anxiolytic effects, but was able to prevent (p<0.01) the UCMS-induced anxiety as assessed by the light/dark test (time spent in the lit area, POEE 100 and 300mg/kg 235.9+/-20.6s and 250.4+/ 17.4s, respectively, compared to DMSO 104.7+/-24.4s). Likewise, although POEE did not induce noticeable effects on glycemia, it effectively (p<0.01) prevented the UCMS-induced hyperglycemia (POEE 100 and 300mg/kg 106.4+/-6.7mg/dl and 107.3+/ 3.3mg/dl, respectively, compared to DMSO 134.6+/-5.9mg/dl). Additionally, POEE (50-200mg/kg i.p. and 800mg/kg p.o.) significantly (p<0.01 and p<0.05, respectively) increased the time to hypoxia-induced convulsion (by 38%, 51%, 59% and 27%, respectively for i.p. and p.o. treatments). The data indicate that POEE counteracts some of the effects brought about by chronic stress. This study combined with the identified antioxidant and neuroprotective properties, as well as the claimed benefits associated with stressful periods suggest that Ptychopetalum olacoides (Marapuama) might possess adaptogen-like properties. PMID- 19682883 TI - Prognostic value of the 6min walk test in bronchiolitis obliterans syndrome. AB - Bronchiolitis Obliterans Syndrome (BOS) complicates the course of many lung transplant recipients. It carries significant risk of morbidity and mortality, but its course is difficult to characterize. We evaluated the prognostic utility of the 6min walk test (6MWT) obtained after the onset of BOS in 42 patients. This was compared to the prognostic significance of changes in the FEV(1). The median time between the onset of BOS and the 6MWT was 109 days. The median decline in the FEV(1) from baseline to BOS onset was 25.7%, while the median change over the ensuing 3 months was 12.5%. Neither of these was predictive of subsequent mortality. The 6MWT yielded averages in the resting saturation, lowest saturation, distance walked and maximal Borg scores of 97%, 90.2%, 323m and 2.35, respectively. The best of these parameters in discriminating survival was the distance. Patients who walked further than 330m had a median survival of 1178 days versus 263 days for those who walked less (p<0.0001). We conclude that the 6MWT provides important prognostic information in patients with BOS and might perform better than spirometry. Use of this test might allow different clinical phenotypes to be discerned. PMID- 19682882 TI - Management of the highly sensitized patient. AB - One of the major challenges in the current era of organ transplantation is to find suitable organs for highly sensitized patients. Different approaches have been successful in a proportion of the patients. Several organ exchange organizations are currently implementing an acceptable mismatch program similar to the one developed by Eurotransplant. The basis of such a program is the accurate definition of those HLA antigens or epitopes toward which the patient did not form antibodies. Donors, who are compatible with the combination of the HLA type of the recipient and these acceptable HLA mismatches, will have a negative crossmatch and are allocated with the highest priority to the highly sensitized patient. Such an approach has shown to increase the transplantation rate with excellent results. Another possibility to find a crossmatch negative donor for highly sensitized patients, who have a positive crossmatch with their potential living donor, is via a (national) paired kidney exchange program. Alternative approaches aim at desensitization of the patient in order to be able to transplant the patient with a donor despite the original crossmatch was positive. Recent protocols include the use of intravenous immunoglobulin in combination with an anti CD20 monoclonal antibody (Rituximab). In order to eliminate the antibody producing plasma cells as well, Bortezomib is recently proposed as a possible agent for future desensitization protocols. As none of these approaches are successful in all patients, a comprehensive use of a combination of these strategies seems the way to go. PMID- 19682884 TI - Enhanced cost-benefit analysis of strategies for LTBI screening and INH chemoprevention in Germany. AB - OBJECTIVES: There is only limited economic data in head-to head comparison between a whole blood QuantiFERON TB Gold in tube (QFT) and the tuberculin skin test (TST) when screening and treating for latent tuberculosis infection (LTBI), and no published study to date that takes into account the predictive value of the two tests. METHODS: Health and economic outcomes of isoniazid preventive treatment (IPT) of close contacts were compared in a decision tree model to perform a cost-benefit analysis with respect to isoniazid related hepatotoxicity and early post-exposure TB over a 2-y period, using the QFT or TST alone or QFT as a confirmatory test for TST results. RESULTS: Cost of screening and treating for using the QFT alone amounted to euro215.79 per close contact, less than that of dual step-testing (euro227.89) or using TST alone (euro232.58). Savings amounted to euro12,200 or euro16,791 per 1000 close contacts, respectively. QFT based procedures were most sensitive to low compliance with IPT or increasing price. Costs of dual step screening was mostly influenced by cost of treating TB disease. When the progression rate for QFT was lowered to that for the TST in a sensitivity analysis, the relationship between the strategies remained robust. In addition, costs of the QFT strategy decreased to euro165.1, and those of the dual step strategy to euro218.4. CONCLUSION: IPT on the basis of using the QFT assay alone produces less cost and reduces more TB cases than other strategies in a low incidence setting. These data have implications for the rational implementation of screening strategies in contact investigation. PMID- 19682885 TI - Postcoital hemoptysis: our experience and review of the literature. AB - Hemoptysis after physical activity is a well-known phenomenon. Hemoptysis following sexual intercourse is rarely reported. We describe three patients with hemoptysis occurring only after sexual activity and not following other types of physical effort. The underlying causes were congestive heart failure, uncontrolled hypertension and Takayasu arteritis. The literature is reviewed. We conclude that hemoptysis can present rarely following sexual activity and is usually associated with cardiovascular decompensation. It is always necessary to search for other underlying diseases. We suggest that patients with unexplained hemoptysis should be specifically asked about postcoital hemoptysis should be included in the differential diagnosis of every patient with unexplained hemoptysis. PMID- 19682886 TI - A randomized controlled trial of the effect of combined spinal-epidural analgesia on the success of external cephalic version for breech presentation. AB - BACKGROUND: Improving the success of external cephalic version (ECV) for breech presentation may help avoid some cesarean deliveries. The results of randomized trials comparing the success of ECV with neuraxial analgesia compared to control are inconsistent. We hypothesized that combined spinal-epidural (CSE) analgesia would increase the success of ECV when compared with systemic opioid analgesia. METHODS: Parturients with singleton breech presentation (n=96) were randomized to receive CSE analgesia with bupivacaine 2.5mg and fentanyl 15 microg (CSE group) or intravenous fentanyl 50 microg (SYS group) before ECV attempt. The primary outcome was ECV success. RESULTS: The success rate of ECV was 47% with CSE and 31% in the SYS group (P=0.14). Subsequent vaginal delivery was 36% for CSE and 25% for SYS (P=0.27). Median [IQR] visual analog pain scores (0-100mm scale) were lower with CSE (3 [0-12]) compared to SYS analgesia (36 [16 to 54]) (P<0.005) and patient satisfaction (0-10 scale) was higher (CSE 10 [9 to 10] versus SYS 7 [4 to 9]) (P<0.005). There were no differences in fetal heart rate patterns, but median time to return to fetal heart rate reactivity after analgesia was shorter with CSE (13 [IQR 9-21] min) compared to the SYS group (39 [IQR 23-51] min) (P=0.02). CONCLUSIONS: There was no difference in the rate of successful ECV or vaginal delivery with CSE compared to intravenous fentanyl analgesia. Pain scores were lower and satisfaction higher with CSE analgesia, and median time to fetal heart rate reactivity was shorter in the CSE group. PMID- 19682887 TI - Engineering G protein-coupled receptors to facilitate their structure determination. AB - Over the last two years, 10 new high-resolution structures of G protein-coupled receptors (GPCRs), either with antagonist bound or in an active-like state, have been solved. Whilst the structures of bovine opsin and squid rhodopsin were determined using protein purified from native sources, a rhodopsin mutant structure, the structures of the beta(1) and beta(2) adrenergic receptors and the adenosine A(2a) receptor were determined from engineered protein heterologously expressed in either insect or mammalian cells. These results are the culmination of years of careful work and have resulted in three new strategies for structure determination of GPCRs that can be applied to virtually any membrane protein. Structural and functional investigations have defined a number of conserved interaction networks between key residues in GPCRs that are probably important for receptor structure and activation. Recent evidence indicates that these networks could be disrupted and rearranged independently from each other, suggesting a possible mechanism for full and partial receptor activation. In addition, one of the opsin structures suggests how one of the highest conserved residues in GPCRs, Arg135(3.50) of the (E/D)RY motif in TM3, interacts directly with a bound peptide derived from the carboxy terminus of the alpha-subunit of the G protein (G(alpha)t). This result sets the basis for the elucidation of the relationship between the conformational changes in the receptor and activation of the G protein. PMID- 19682888 TI - Lipopeptide detergents for membrane protein studies. AB - An ideal detergent would be able to maintain a membrane protein in a soluble state with no measurable effect on the functional, structural, and thermodynamic properties of the protein relative to the bilayer-embedded state. Unfortunately, the detergents that are commonly used by membrane protein biochemists fall short of this standard. Although remarkable advances have been made in membrane protein structural biology, there remains a need for improved detergents that provide a more natural substitute for the membrane environment. Lipopeptide detergents (LPDs) are a new class of amphiphile designed to be better mimics of the bilayer at the hydrophobic surfaces of solubilized membrane proteins. LPDs consist of an alpha-helical peptide backbone that supports alkyl chains anchored at either end of the helix. The LPD monomers self-assemble into cylindrical micelles with a membrane-like packing of the inner core of alkyl chains. PMID- 19682890 TI - Defining a high-risk subgroup with colon cancer stages I and II for possible adjuvant therapy. AB - AIM: Adjuvant therapy is not routinely recommended in UICC stages I and II colon cancer, but may be considered for high-risk patients. Our aim is to identify clinicopathologic characteristics in colon cancer stages I and II, which are associated with an increased risk of tumour recurrence and tumour-related death. METHODS: We analysed our prospectively documented clinical database of 775 patients with colon cancer stages I and II, which underwent curative resection between 1982 and 2006. No adjuvant chemotherapy was applied. The median follow-up time was 80 months. RESULTS: For the entire study group, 5- and 10-year tumour specific survival probabilities were 94.8+/-0.9% and 91.0+/-1.4%, respectively. Multivariate analysis identified three tumour characteristics as independent prognostic factors: lymphatic vessel invasion (p=0.034), poor tumour grading (G3/G4) (p=0.020) and extended tumour length (6 cm) (p=0.042). Five-year (10 year) tumour-specific survival for patients without any of the poor prognostic tumour characteristics (ppTCs) was 96.0% (94.7%). There was a significantly increased risk for tumour-related death with increasing numbers of ppTCs (p<0.001). While patients with only one ppTC had a 5-year (10-year) tumour specific survival of 94.8% (88.9%), it decreased to 88.9% (78.4%) for patients with two ppTCs (hazard ratio (HR) 3.69, 95% confidence interval (CI) 1.67-8.13) and to 87.5% (72.9%) for patients with all three ppTCs (HR 6.56, 95% CI 1.50 26.62). CONCLUSION: Patients with stage I or II colon cancer have a favourable prognosis after radical resection. The presence of two or three poor prognostic tumour characteristics identifies a small patient subgroup (12%) with an increased risk of tumour-related death that may be considered for adjuvant chemotherapy. PMID- 19682889 TI - Acute toxicity of curative radiotherapy for intermediate- and high-risk localised prostate cancer in the EORTC trial 22991. AB - INTRODUCTION: This trial randomly assessed short-term adjuvant hormonal therapy added to radiotherapy (RT) for intermediate- and high-risk (UICC 1997 cT2a or cT1b-c with high PSA or Gleason score) localised prostate cancer. We report acute toxicity (CTCAE v2) assessed weekly during RT in relation to radiation parameters. PATIENTS AND METHODS: Centres selected the RT dose (70, 74 or 78Gy) and RT technique. Statistical significance is at 0.05. RESULTS: Of 791 patients, 652 received 3D-CRT (70Gy: 195, 74Gy: 376, 78Gy: 81) and 139 received IMRT (74Gy: 28, 78Gy: 111). During RT, grade 3 gastrointestinal (GI) and genitourinary (GU) toxicities were reported by 7 (0.8%) and 50 (6.3%) patients, respectively. No grade 4 was reported. The risk of grade 2 GI toxicity increased significantly with increasing D50%-rectum (p=0.004) and that of grade 2 GU toxicity correlated only to Dmax-bladder (p=0.051). 3D-RT technique, increasing total dose and V95% >400 cc increased D50% and Dmax. One month after RT, only 14 patients (1.8%) reported grade 3 toxicity. AST did not seem to influence the risk of GU or GI acute toxicity. CONCLUSION: RT up to 78Gy was well tolerated. Dmax-bladder and D50%-rectum influenced the risk of grade 2 GU toxicity and GI toxicity, respectively. Both were lower with IMRT but remained high for an irradiated RT volume>400 cc for 3D-RT and for a dose of 78Gy. Hormonal treatment did not influence acute toxicity. PMID- 19682891 TI - Improved survival among younger but not among older patients with Multiple Myeloma in the Netherlands, a population-based study since 1989. AB - This study assesses whether new treatment strategies developed in clinical trials translate into improved survival for multiple myeloma (MM) patients in the Netherlands. All patients diagnosed with MM in the Northern part of the Netherlands between 1989 and 2005 were retrieved from two regional population based cancer registries. Information on study participation was derived from linkage with trial information systems. The effect of period of diagnosis (1989 1992, 1993-1996, 1997-2000, 2001-2005), age (<50, 50-65, 66-74, 75), gender, Salmon-Durie (SD) stage, trial participation and treatment on relative survival were studied. In total 4985 patients were included. When trial participation was analysed for exact periods in which trials were open, 16% of patients aged 65 years with SD-stage I and 38% with SD-stage II or III were enrolled compared to 2% of patients aged >65 years with SD-stage I and 5% with SD-stage II or III. Relative survival decreased with age (p<.001), with advanced stage (p<.001) and was better for patients enrolled in trials (p<.001). Five-year relative survival increased from 34% (95% confidence interval (95% CI) 28-39%) in 1989-1992 to 56% (95% CI 50-61%) in 2001-2005 for patients 65 years. The excess mortality was 37% lower in 2001-2005 than in 1989-1992 for these patients, adjusted for age, stage, trial participation and gender (p<.001). Survival did not improve for older patients. IN CONCLUSION: MM survival improved among younger but not among older patients since the mid-1990s. The improved survival of younger patients coincided with increasing trial participation and increasing use of high-dose chemotherapy and autologous stem-cell transplantation. PMID- 19682892 TI - Hybrid process of BAC and sMBR for treating polluted raw water. AB - The hybrid process of biological activated carbon (BAC) and submerged membrane bioreactor (sMBR) was evaluated for the drinking water treatment from polluted raw water, with the respective hydraulic retention time of 0.5 h. The results confirmed the synergetic effects between the BAC and the subsequent sMBR. A moderate amount of ammonium (54.5%) was decreased in the BAC; while the total removal efficiency was increased to 89.8% after the further treatment by the sMBR. In the hybrid process, adsorption of granular activated carbon (in BAC), two stages of biodegradation (in BAC and sMBR), and separation by the membrane (in sMBR) jointly contributed to the removal of organic matter. As a result, the hybrid process managed to eliminate influent DOC, UV(254), COD(Mn), TOC, BDOC and AOC by 26.3%, 29.9%, 22.8%, 27.8%, 57.2% and 49.3%, respectively. Due to the pre treatment effect of BAC, the membrane fouling in the downstream sMBR was substantially mitigated. PMID- 19682893 TI - The preparation of High caloric fuel (HCF) from water hyacinth by deoxy liquefaction. AB - The aim of this study was to prepare HCF (High caloric fuel) from WH (water hyacinth) by deoxy-liquefaction and investigate the composition of HCF. The experiment was performed in a closed reactor at different final temperatures (573 K, 623 K, 673 K and 723 K) with the heating rate of 60 K/min. At 623 K, the maximum yield (12.6 wt.%) of HCF with heating value of 43.8 MJ/kg was obtained. Alkanes, benzene derivatives and phenol derivatives were the dominant components in HCF. The main component in gaseous product was CO(2) (93.2 mol%), which meant that the oxygen in WH was released mainly in the form of CO(2). In addition, the elemental analysis of solid char suggested that the residue content of hydrogen was too low to produce more HCF. So, deoxy-liquefaction mentioned in this article was an effective way to remove oxygen and utilize C and H in WH to the greatest extent. PMID- 19682894 TI - Photoregulation of thrombin aptamer activity using Bhc caging strategy. AB - Thrombin aptamer was attempted to cage with Bhc (6-bromo-7-hydroxycoumarin-4 ylmethyl) group for controlling its specific affinity to target molecular through photolysis. By multiple-caging strategy, aptamer could be rendered biologically inert and partially restored with subsequently illumination. This provides a convenient method for photoregulating aptamer activity with exact spatiotemporal resolution. PMID- 19682895 TI - Synthesis of novel [1,2]-diamines with antituberculosis activity. AB - Guided by the metabolism information of SQ109, derivatives with substituted geranylamine moiety or substituted admantane ring of SQ109 were synthesized and evaluated as antituberculosis agents. Among all tested compounds, compound 11c showed the most potent antituberculosis activity with MIC value of 0.3 microM against Mycobacterium tuberculosis H37Rv. PMID- 19682896 TI - C-5 Substituted heteroaryl 3-pyridinecarbonitriles as PKCtheta inhibitors: Part I. AB - We earlier reported that 3-pyridinecarbonitriiles with a 4-methylindolyl-5-amino group at C-4 and a phenyl group at C-5 were inhibitors of PKCtheta. Keeping the group at C-4 of the pyridine core constant, we varied the water solubilizing group on the phenyl ring at C-5 and then replaced the C-5 phenyl ring with several monocyclic heteroaryl rings, including furan, thiophene and pyridine. Analog 6e with a 4-methylindol-5-ylamino group at C-4 and a 5-[(4-methylpiperazin 1-yl)methyl]-2-furyl group C-5 had an IC50 value of 4.5 nM for the inhibition of PKCtheta. PMID- 19682897 TI - Water-soluble phosphate prodrugs of pleuromutilin analogues with potent in vivo antibacterial activity against Gram-positive pathogens. AB - A phosphate prodrug strategy was investigated to address the problem of poor aqueous solubility of pleuromutilin analogues. Water-soluble phosphate prodrugs 6a, 6b and 6c of pleuromutilin analogues were designed and synthesized. Three compounds all exhibited excellent aqueous solubility (>50mg/mL) at near-neutral pH and sufficient stability in buffer solution. In particular, the phenol pleuromutilin prodrug 6c displayed favourable pharmacokinetic profiles and comparable potency with vancomycin against MSSA and MRSA strains in vivo. PMID- 19682898 TI - Discovery of 4H-pyrazolo[1,5-a]pyrimidin-7-ones as potent inhibitors of hepatitis C virus polymerase. AB - A series of 4H-pyrazolo[1,5-a]pyrimidin-7-one derivatives was synthesized and evaluated for inhibitory activity against HCV NS5B RNA-dependent RNA polymerase. A number of these compounds exhibited potent activity in enzymatic assay. The synthesis and structure-activity relationship are also described. PMID- 19682899 TI - Discovery of 3,3-disubstituted piperidine-derived trisubstituted ureas as highly potent soluble epoxide hydrolase inhibitors. AB - 3,3-Disubstituted piperidine-derived trisubstituted urea entA-2b was discovered as a highly potent and selective soluble epoxide hydrolase (sEH) inhibitor. Despite the good compound oral exposure, excellent sEH inhibition in whole blood, and remarkable selectivity, compound entA-2b failed to lower blood pressure acutely in spontaneously hypertensive rats (SHRs). This observation further challenges the premise that sEH inhibition can provide a viable approach to the treatment of hypertensive patients. PMID- 19682900 TI - 2,3,4,5-Tetrahydro-1H-pyrido[2,3-b and e][1,4]diazepines as inhibitors of the bacterial enoyl ACP reductase, FabI. AB - In the search for new antibacterial agents, the enzyme FabI has been identified as an attractive target. Employing a structure guided approach, the previously reported ene-amide series of FabI inhibitors were expanded to include 2,3,4,5 tetrahydro-1H-pyrido[2,3-b and e][1,4]diazepines. These novel series incorporate additional H-bonding functions and can be more water soluble than their naphthyridinone progenitors; diazepine 16c is shown to be efficacious in a mouse infection model. PMID- 19682901 TI - Spiro-naphthyridinone piperidines as inhibitors of S. aureus and E. coli enoyl ACP reductase (FabI). AB - Spiropiperidine naphthyridinone inhibitors of Staphylococcus aureus and Escherichia coli FabI have been prepared. Compounds 14a and 14c were identified as having sub-nanomolar E. coli FabI activity and are among the most potent FabI inhibitors yet described. The structural model of 14a bound to E. coli FabI is shown. PMID- 19682902 TI - Antimicrobial dendrimer active against Escherichia coli biofilms. AB - We have investigated the ability of a previously reported antimicrobial peptide dendrimer (RW)(4D) to inactivate Escherichia coli RP437 in planktonic culture and in biofilms. The results show that the dendrimer inhibits bacterial growth in both planktonic and biofilm states. Live/Dead staining assays reveal that most bacteria in a preformed biofilm lose viability after treatment with this peptide. This result is in marked contrast to most existing reports that antimicrobial peptides are ineffective against mature bacterial biofilms. PMID- 19682903 TI - Outcome of T1N0M0 breast cancer in relation to St. Gallen risk assignment criteria for adjuvant therapy. AB - T1N0M0 (stage I) breast cancer (BC) has been increasing in recent decades but the optimal adjuvant approach remains controversial. To assess the outcome of BC patients stratified and treated with multimodal therapies according to St. Gallen consensus meeting recommendations, we retrospectively evaluated an unselected cohort of T1N0M0 BC patients, with respect to the St. Gallen criteria. At a median follow-up of 5 years, the recurrence rate, recurrence-free survival and overall survival were 7%, 94% and 96% respectively, and 60% of relapses were locoregional. No statistically significant difference was observed between T1a,b/T1c groups, or among risk categories (high/intermediate/low). The very low rate of distant recurrences even in patients with unfavorable prognostic factors seems to support the use of adjuvant systemic therapies but better prognostic and predictive factors are strongly needed for this subset of patients. PMID- 19682904 TI - Cell-size-dependent spindle elongation in the Caenorhabditis elegans early embryo. AB - Cell size is one of the critical parameters controlling the size of intracellular structures. A well-known example is the constant nuclear-to-cytoplasmic ratio (N/C ratio) [1-5]. The length of the metaphase spindle is proportional to cell size, but it has an upper limit during early embryogenesis [6]. During anaphase, the mitotic spindle elongates and delivers the centrosomes and sister chromatids near the centers of the nascent daughter cells. Here, we quantified the relationship between spindle elongation and cell size in the early embryo of Caenorhabditis elegans and propose possible models for cell-size-dependent spindle elongation. Quantitative measurements revealed that the extent and speed of spindle elongation are correlated with cell size throughout early embryogenesis. RNAi knockdown of Galpha proteins and their regulators revealed that the spindles failed to fully elongate and that the speed of spindle elongation was almost constant regardless of cell size. Our results suggest that spindle elongation is controlled by two qualitatively distinct mechanisms, i.e., Galpha-dependent and -independent modes of elongation. Simulation analyses revealed that the constant-pulling model and the force-generator-limited model reproduced the dynamics of the Galpha-independent and Galpha-dependent mechanisms, respectively. These models also explain how the set length of spindles is achieved. PMID- 19682905 TI - Dissociation of neural mechanisms underlying orientation processing in humans. AB - Orientation selectivity is a fundamental, emergent property of neurons in early visual cortex, and the discovery of that property has dramatically shaped how we conceptualize visual processing. However, much remains unknown about the neural substrates of this basic building block of perception, and what is known primarily stems from animal physiology studies. To probe the neural concomitants of orientation processing in humans, we employed repetitive transcranial magnetic stimulation (rTMS), which can significantly attenuate neuronal spiking activity, hemodynamic responses, and local field potentials within a focused cortical region. Using rTMS to suppress neural responses evoked by stimuli falling within a local region of the visual field, we were able to dissociate two distinct components of the neural circuitry underlying orientation processing: selectivity and contextual effects. Orientation selectivity gauged by masking was unchanged by rTMS, whereas an otherwise robust orientation repulsion illusion was weakened after rTMS. This dissociation implies that orientation processing in humans relies on distinct mechanisms, only one of which was impacted by rTMS. These results are consistent with models positing that orientation selectivity is governed by patterns of convergence of thalamic afferents onto cortical neurons, with intracortical activity then shaping population responses amongst those cortical neurons. PMID- 19682906 TI - Reprogramming after chromosome transfer into mouse blastomeres. AB - It is well known that oocytes can reprogram differentiated cells, allowing animal cloning by nuclear transfer. We have recently shown that fertilized zygotes retain reprogramming activities, suggesting that such activities might also persist in cleavage-stage embryos. Here, we used chromosome transplantation techniques to investigate whether the blastomeres of two-cell-stage mouse embryos can reprogram more differentiated cells. When chromosomes from one of the two blastomeres were replaced with the chromosomes of an embryonic or CD4(+) T lymphocyte donor cell, we observed nuclear reprogramming and efficient contribution of the manipulated cell to the developing blastocyst. Embryos produced by this method could be used to derive stem cell lines and also developed to term, generating mosaic "cloned" animals. These results demonstrate that blastomeres retain reprogramming activities and support the notion that discarded human preimplantation embryos may be useful recipients for the production of genetically tailored human embryonic stem cell lines. PMID- 19682907 TI - Cultural confusions show that facial expressions are not universal. AB - Central to all human interaction is the mutual understanding of emotions, achieved primarily by a set of biologically rooted social signals evolved for this purpose-facial expressions of emotion. Although facial expressions are widely considered to be the universal language of emotion, some negative facial expressions consistently elicit lower recognition levels among Eastern compared to Western groups (see [4] for a meta-analysis and [5, 6] for review). Here, focusing on the decoding of facial expression signals, we merge behavioral and computational analyses with novel spatiotemporal analyses of eye movements, showing that Eastern observers use a culture-specific decoding strategy that is inadequate to reliably distinguish universal facial expressions of "fear" and "disgust." Rather than distributing their fixations evenly across the face as Westerners do, Eastern observers persistently fixate the eye region. Using a model information sampler, we demonstrate that by persistently fixating the eyes, Eastern observers sample ambiguous information, thus causing significant confusion. Our results question the universality of human facial expressions of emotion, highlighting their true complexity, with critical consequences for cross cultural communication and globalization. PMID- 19682908 TI - Centriole age underlies asynchronous primary cilium growth in mammalian cells. AB - Primary cilia are microtubule-based sensory organelles that play important roles in development and disease . They are required for Sonic hedgehog (Shh) and platelet-derived growth factor (PDGF) signaling. Primary cilia grow from the older of the two centrioles of the centrosome, referred to as the mother centriole. In cycling cells, the cilium typically grows in G1 and is lost before mitosis, but the regulation of its growth is poorly understood. Centriole duplication at G1/S results in two centrosomes, one with an older mother centriole and one with a new mother centriole, that are segregated in mitosis. Here we report that primary cilia grow asynchronously in sister cells resulting from a mitotic division and that the sister cell receiving the older mother centriole usually grows a primary cilium first. We also show that the signaling proteins inversin and PDGFRalpha localize asynchronously to sister cell primary cilia and that sister cells respond asymmetrically to Shh. These results suggest that the segregation of differently aged mother centrioles, an asymmetry inherent to every animal cell division, can influence the ability of sister cells to respond to environmental signals, potentially altering the behavior or fate of one or both sister cells. PMID- 19682909 TI - Rehabilitation results following anterior cruciate ligament reconstruction using a hard brace compared to a fluid-filled soft brace. AB - The purpose of this study was to compare the clinical outcomes of rehabilitation after ACL reconstruction using a water-filled soft brace to those using a hard brace. The method used in this study was a prospective randomised clinical trial including 36 patients wearing a hard brace and 37 patients wearing a water-filled soft brace for 6 weeks after surgery. Preoperative and postoperative (seven examinations) clinical evaluation within a follow-up period of 1 year including effusion status, swelling and range of motion (ROM), IKDC 2000, KT1000 Arthrometer, Lysholm knee scoring scale and Tegner activity score. Mean values are presented with standard deviations. Data was analysed using descriptive statistics and Student's t-test for unpaired samples. Significantly less effusion was found in the soft brace group from 5 days (p=0.002) to 12 weeks (p<0.024) postoperative. Hard brace patients presented with significantly more extension deficit from 5 days (p=0.036) to 12 months (p=0.014) postoperative but no significant difference was detected in complete ROM, laxity or thigh atrophy at any follow-up examination. Patients treated with a soft brace had significantly higher IKDC subjective ratings at 6 weeks (p=0.02) up to 12 months after operation (p=0.002) and rated significantly higher in Tegner activity score (p=0.004) and Lysholm knee scoring scale (p=0.006) 6 and 12 months (p<0.001 for both scores) postoperatively. The water-filled soft brace was superior regarding effusion, swelling, extension deficit and patient-measured midterm outcome. The soft brace presents a safe, easy-to-use and effective alternative to the hard brace. PMID- 19682910 TI - Synthesis and molecular modeling of some novel hexahydroindazole derivatives as potent monoamine oxidase inhibitors. AB - A novel series of 2-thiocarbamoyl-2,3,4,5,6,7-hexahydro-1H-indazole and 2 substituted thiocarbamoyl-3,3a,4,5,6,7-hexahydro-2H-indazoles derivatives were synthesized and investigated for the ability to inhibit the activity of the A and B isoforms of monoamine oxidase (MAO). The target molecules were identified on the basis of satisfactory analytical and spectra data (IR, (1)H NMR, (13)C NMR, (2)D NMR, DEPT, EI-MASS techniques and elemental analysis). Synthesized compounds showed high activity against both the MAO-A (compounds 1d, 1e, 2c, 2d, 2e) and the MAO-B (compounds 1a, 1b, 1c, 2a, 2b) isoforms. In the discussion of the results, the influence of the structure on the biological activity of the prepared compounds was delineated. It was suggested that non-substituted and N methyl/ethyl bearing compounds (except 2c) increased the inhibitory effect and selectivity toward MAO-B. The rest of the compounds, carrying N-allyl and N phenyl, appeared to select the MAO-A isoform. The inhibition profile was found to be competitive and reversible for all compounds. A series of experimentally tested (1a-2e) compounds was docked computationally to the active site of the MAO A and MAO-B isoenzyme. The autodock 4.01 program was employed to perform automated molecular docking. In order to see the detailed interactions of the docked poses of the model inhibitors compounds 1a, 1d, 1e and 2e were chosen because of their ability to reversibly inhibit the MAO-B and MAO-A and the availability of experimental inhibition data. The differences in the intermolecular hydrophobic and H-bonding of ligands to the active site of each MAO isoform were correlated to their biological data. Observation of the docked positions of these ligands revealed interactions with many residues previously reported to have an effect on the inhibition of the enzyme. Excellent to good correlations between the calculated and experimental K(i) values were obtained. In the docking of the MAO-A complex, the trans configuration of compound 1e made various very close interactions with the residues lining the active site cavity these interactions were much better than those of the other compounds tested in this study. This tight binding observation may be responsible for the nanomolar inhibition of form of MAOA. However, it binds slightly weaker (experimental K(i)=1.23 microM) to MAO-B than to MAO-A (experimental K(i)=4.22 nM). PMID- 19682911 TI - Synthesis and structure-activity relationships of N-aryl(indol-3-yl)glyoxamides as antitumor agents. AB - The synthesis and study of the structure-activity relationships of cytotoxic compounds based on N-pyridinyl or N-aryl-2-(1-benzylindol-3-yl)glyoxamide skeleton, represented by the lead structures D-24241 and D-24851, are described. The presence of N-(pyridin-4-yl) moiety was crucial for activity and 2-[1-(4 chloro-3-nitrobenzyl)-1H-indol-3-yl]-2-oxo-N-(pyridin-4-yl)acetamide (55), the most potent derivative, showed IC(50)=39 nM, 51 nM and 11 nM against HeLa/KB (human cervix carcinoma), L1210 (murine leukemia) and SKOV3 (human ovarian carcinoma) cell lines proliferation assay, respectively, as active as the lead compounds. PMID- 19682912 TI - Synthesis and pharmacological evaluation of novel 1- and 8-substituted-3-furfuryl xanthines as adenosine receptor antagonists. AB - The synthesis of an important set of 3-furfurylxanthine derivatives is described. Binding affinities were determined for rat A(1) and human A(2A), A(2B) and A(3) receptors. Several of the 3-furfuryl-7-methylxanthine derivatives showed moderate to-high affinity at human A(2B) receptors, the most active compound (10d) having a K(i) of 7.4 nM for hA(2B) receptors, with selectivities over rA(1) and hA(2A) receptors up to 14-fold and 11-fold, respectively. Affinities for hA(3) receptors were very low for all members of the set. PMID- 19682913 TI - Death receptor 5 promoter-enhancing compounds isolated from Catimbium speciosum and their enhancement effect on TRAIL-induced apoptosis. AB - The TRAIL/death-receptor signaling pathway has been considered a promising target for selective cancer therapy, although some malignant tumors exhibit TRAIL resistance. We previously found that isoflavonoid enhanced TRAIL-induced apoptosis in TRAIL-resistant cells, which is achieved through up-regulation of death receptor 5 (DR5). In our screening program targeting DR5 promoter enhancement activity, activity-guided fractionations of the extract of Catimbium speciosum led to the isolation of six compounds. Of the isolates, cardamomin (6), the most potent compound, enhanced the expressions of DR5 and DR4 and decreased the Bcl-xL level in TRAIL-resistant DLD1 cells. The combination of 6 and TRAIL synergistically enhanced TRAIL-induced apoptosis against TRAIL-resistant cells upon the activation of caspase-8, 9, and 3. In addition, enhancement of apoptosis by 6 was inhibited by human recombinant DR5/Fc and DR4/Fc chimera proteins, TRAIL neutralizing fusion proteins, indicating that 6 sensitize TRAIL-resistant cells to TRAIL through the induction of DR5 and DR4. Also, up-regulation of DR5 by 6 paralleled that of CCAAT/enhancer-binding protein-homologous protein (CHOP). PMID- 19682915 TI - New correction method for dynamic error in on-line gamma ray thickness detection. AB - In this paper, we present a new method to correct dynamic error (DE) in on-line gamma ray thickness measurement, which significantly improves measurement precision over traditional method, in most cases, by one order of magnitude. Theoretical analysis of DE is presented and the correction method is proposed. In order to further prove our theory, Monte Carlo simulation is taken and the performance improvement is given. The method has been successfully applied to our thickness measurement system and brought dramatic improvement to its dynamic precision. PMID- 19682916 TI - Six year follow-up of a patient treated with mandibular symphyseal distraction osteogenesis. AB - In this report, treatment and long term effects of mandibular symphyseal distraction osteogenesis (MSDO) on dentofacial structures are presented. The distraction device used in the present case was a custom made tooth- and bone borne distractor. Post-treatment records revealed elimination of mandibular anterior crowding, no temporomandibular joint (TMJ) disorder, no periodontal pathology and no root resorption. Follow-up records taken 6 years after the completion of the treatment indicated no mandibular anterior crowding or TMJ disorder. Treatment and long term results of this case suggest that mandibular widening with distraction osteogenesis is an efficient, safe and stable treatment modality for patients presenting transverse mandibular deficiency and/or mandibular anterior dental crowding. However, large sample studies are required to draw substantial conclusions about the long term effects of the MSDO on dentofacial structures. PMID- 19682917 TI - Routine placement of an intercostal chest drain during video-assisted thoracoscopic surgical lung biopsy unnecessarily prolongs in-hospital length of stay in selected patients. AB - OBJECTIVE: Video-assisted thoracoscopic surgical (VATS) lung biopsy is frequently used in the diagnosis of parenchymal lung disease. However, there is still debate over the need for routine use of an intercostal chest drain after this procedure. This study aimed to evaluate the necessity of positioning an intercostal chest drain as an integral part of VATS lung biopsy. METHODS: Data from VATS lung biopsies performed over a 5-year period were retrospectively analysed. Patients in whom there was evidence of air leak intra-operatively following lung biopsy were excluded. Patients in whom no air leak was detected on testing were included in this study. A chest drain was inserted solely according to the surgeons' practice. RESULTS: This study included 175 patients. Of these, 82 patients had an intercostal chest drain positioned during the VATS procedure and 93 did not. There were no significant differences between the two groups in terms of mean (standard deviation (SD)), age (54.4 (14.9) vs 55.8 (13.5) years, p=0.58), gender (63% vs 59% males, p=0.56) or side of procedure (45% vs 56% right side, p=0.22). One patient in the 'no drain' group developed a clinically significant pneumothorax 24h after surgery and required a drain to be inserted. There was also no significant difference between the two groups in the incidence of radiologically detected pneumothorax immediately post-procedure (23% vs 20%, p=0.66) or on postoperative day 1 (26% vs 20%, p=0.63). There was no significant difference in the incidence of pneumothorax on follow-up (at 4-6 weeks) chest radiograph (10% vs 7%, p=0.61). In all cases, the pneumothoraces were small and not clinically significant. However, there was a significant difference in the median (inter-quartile range (IQR)) length of stay between the two groups (3 (2,4) vs 2 (1,3) days, respectively, p<0.001). CONCLUSIONS: The routine use of an intercostal chest drain after VATS lung biopsy unnecessarily increases the length of hospital stay without reduction in the incidence of pneumothorax. PMID- 19682914 TI - Inhibition of S-ribosylhomocysteinase (LuxS) by substrate analogues modified at the ribosyl C-3 position. AB - S-ribosylhomocysteinase (LuxS) catalyzes the cleavage of the thioether bond of S ribosylhomocysteine (SRH) to produce homocysteine and 4,5-dihydroxy-2,3 pentanedione (DPD), which is the precursor of type 2 autoinducer for bacterial cell-cell communication. In this work, we have synthesized several SRH analogues modified at the ribose C3 position as potential inhibitors of LuxS. While removal or methylation of the C3-OH resulted in simple competitive inhibitors of moderate potency, inversion of the C3 stereochemistry or substitution of fluorine for C3 OH resulted in slow-binding inhibitors of improved potency. The most potent inhibitor showed a K(I)(*) value of 0.43 microM. PMID- 19682918 TI - Transapical implantation of a self-expanding aortic valve bioprosthesis--animal feasibility study. AB - BACKGROUND: Severe peripheral arterial disease may pose a limitation to the applicability of trans-arterial aortic valve implantation in patients who are otherwise candidates. For this reason, transapical aortic valve implantation has been proposed as a possible alternative. OBJECTIVE: To evaluate the acute safety and performance of a specially designed delivery system, the CoreValve Tranzap delivery catheter, for the transapical implantation of a self-expandable aortic valve prosthesis in a porcine animal model. METHODS: Thirteen pigs were implanted with a self-expandable aortic valve bioprosthesis using a 21F catheter delivery system through a transapical approach. The delivery system was evaluated for: (1) the ability to access the implantation site; (2) the ability to precisely position the delivery catheter; (3) control of the delivery of the prosthesis; (4) safe retrieval of the delivery catheter; and (5) the ability to close the apical access site of the heart. RESULTS: Successful implantation was achieved in 100% of the cases. The following points were achieved in all animals: (1) passage of the delivery catheter through an incision in the left ventricular apex; (2) positioning of the delivery catheter on the implantation site; (3) controlled deployment of the aortic valve prosthesis; (4) the safe retrieval of the delivery catheter system; and (5) the adequate closure of the apex of the heart. CONCLUSION: This study demonstrates the acute safety and feasibility of the CoreValve Tranzap delivery system for the transapical implantation of the CoreValve self-expanding aortic valve bioprosthesis in a porcine animal model. PMID- 19682919 TI - Growth suppressive cytokines and the AKT/mTOR pathway. AB - The mTOR signaling pathway plays a very important role in the transmission of signals for initiation of mRNA translation and protein expression in mammalian cells. mTOR activates various downstream effectors to promote initiation of cap dependent mRNA translation and mediate pro-mitogenic and pro-survival signals. Recent evidence has implicated effectors of this signaling cascade in mRNA translation for interferon stimulated genes (ISGs). In addition, it was recently shown that AKT/mTOR-mediated signals play important roles in the generation of IFN-dependent antiviral and growth inhibitory responses, suggesting that mTOR and its effectors can mediate diverse biological responses, depending on the cellular context and the triggering stimuli. In this review, the regulatory effects of various growth suppressive cytokines on the mTOR pathway are summarized and the emerging new functions of mTOR are discussed. PMID- 19682920 TI - Interferon-gamma and interleukin-4 single nucleotide gene polymorphisms in Paracoccidioidomycosis. AB - The gene polymorphisms interferon-gamma (IFN-gamma) +874 T/A and interleukin (IL) 4 -590 C/T have been associated with the altered production of cytokines. Therefore, they might be indicative of the occurrence of Paracoccidioidomycosis (PCM) caused by Paracoccidioides brasiliensis. The analysis of single nucleotide polymorphism (SNP) at position+874 IFN-gamma showed an increase occurrence of A/T genotype in both PCM patients and healthy individuals as control (HIC) (56% and 45%, respectively), while the allelic distribution showed 82% of A allele in the patients and 80% in the controls. The SNP of -590 IL-4 showed that C/T genotype was significantly (p<0.05) more prevalent (39%) in PCM group compared to the HIC group (19%), while IL-4 C/C genotype was significantly less frequent (59%) in the patient group compared to the control group (81%). Otherwise, 41% of PCM patients and 19% of HIC individuals carried the IL-4 T allele. Stimulation of peripheral blood mononuclear cells (PBMC) from PCM patients with cell extract antigenic preparations (PbAg) as well as secreted and surface antigens (MEXO) of P. brasiliensis evidenced that there is no difference in the IFN-gamma production related to A and T alleles between PCM and HIC individuals. However, with IL-4 production, PCM patients classified as C phenotype showed two times more IL-4 production than PCM patients classified as T phenotype and HIC controls. In conclusion, our results suggest that functional genetic variants in the IL-4 promoter could influence the production of IL-4 in PCM. PMID- 19682921 TI - Silver-coated gold nanoparticles as concentrating probes and matrices for surface assisted laser desorption/ionization mass spectrometric analysis of aminoglycosides. AB - A novel method for the determination of aminoglycosides by surface-assisted laser desorption/ionization mass spectrometry (SALDI MS) with the aid of silver-coated gold nanoparticles (Au@AgNPs) has been developed. The Au@AgNPs with surface capped by anionic citrate were used as concentrating probes as well as matrices in SALDI MS. Adsorption of aminoglycosides onto the nanoparticles was mainly through electrostatic attraction. The aminoglycoside-adsorbed nanoparticles were directly characterized by SALDI MS after a simple washing. Using Au@AgNPs to preconcentrate the aminoglycosides from 500 microL buffer solution, the limits of detection (LODs) at signal-to-noise ratio of 3 were 3, 25, 15, 30, and 38 nM for paromomycin, kanamycin A, neomycin, gentamicin, and apramycin, respectively. This method was successfully applied to the determination of aminoglycosides in human plasma samples. The LODs of aminoglycosides in plasma samples were 9, 130, 81, and 180 nM for paromomycin, kanamycin A, neomycin, and gentamicin, respectively. Recoveries of aminoglycosides in plasma samples were about 80%. PMID- 19682922 TI - Ionization mechanism of the ambient pressure pyroelectric ion source (APPIS) and its applications to chemical nerve agent detection. AB - We present studies of the ionization mechanism operative in the ambient pressure pyroelectric ionization source (APPIS), along with applications that include detection of simulants for chemical nerve agents. It is found that ionization by APPIS occurs in the gas-phase. As the crystal is thermally cycled over a narrow temperature range, electrical discharges near the surface of the crystal produce energetic species which, through reactions with atmospheric molecules, result in reactant ions such as protonated water clusters or clusters of hydroxide and water. Reactant ions can be observed directly in the mass spectrometer. These go on to react with trace neutrals via proton transfer reactions to produce the ions observed in mass spectra, which are usually singly protonated or deprotonated species. Further implicating gas-phase ionization, observed product distributions are highly dependent on the composition of ambient gases, especially the concentration of water vapor and oxygen surrounding the source. For example, basic species such as triethylamine are observed as singly protonated cations at a water partial pressure of 10 torr. At a water pressure of 4 torr, reactive oxygen species are formed and lead to observation of protonated amine oxides. The ability of the APPIS source to detect basic molecules with high proton affinities makes it highly suited for the detection of chemical nerve agents. We demonstrate this application using simulants corresponding to VX and GA (Tabun). With the present source configuration pyridine is detected readily at a concentration of 4 ppm, indicating ultimate sensitivity in the high ppb range. PMID- 19682923 TI - Origin of supercharging in electrospray ionization of noncovalent complexes from aqueous solution. AB - The use of m-nitrobenzyl alcohol (m-NBA) to enhance charging of noncovalent complexes formed by electrospray ionization from aqueous solutions was investigated. Addition of up to 1% m-NBA can result in a significant increase in the average charging of complexes, ranging from approximately 13% for the homo heptamer of NtrC4-RC (317 kDa; maximum charge state increases from 42+ to 44+) to approximately 49% for myoglobin (17.6 kDa; maximum charge state increases from 9+ to 16+). Charge state distributions of larger complexes obtained from heated solutions to which no m-NBA was added are remarkably similar to those containing small amounts of m-NBA. Dissociation of the complexes through identical channels both upon addition of higher concentrations of m-NBA and heating is observed. These results indicate that the enhanced charging upon addition of m-NBA to aqueous electrospray solutions is a result of droplet heating owing to the high boiling point of m-NBA, which results in a change in the higher-order structure and/or dissociation of the complexes. For monomeric proteins and small complexes, the enhancement of charging is lower for heated aqueous solutions than from solutions with m-NBA because rapid folding of proteins from heated solutions that do not contain m-NBA can occur after the electrospray droplet is formed and is evaporatively cooled. PMID- 19682924 TI - Ion trap mass analysis at high pressure: a theoretical view. AB - The mass-selective manipulation of ions at elevated pressure, including mass analysis, ion isolation, or excitation, is of great interest for the development of mass spectrometry instrumentation, particularly for systems in which ion traps are employed as mass analyzers or storage devices. While experimental exploration of high-pressure mass analysis is limited by various difficulties, such as ion detection or electrical discharge at high-pressure, theoretical methods have been developed in this work to study ion/neutral collision effects within quadrupole ion traps and to explore their performance at pressures up to 1 Torr. Ion trapping, isolation, excitation, and resonance ejection were investigated over a wide pressure range. The theoretically calculated data were compared with available experimental data for pressures up to 50 mTorr, allowing the prediction of ion trap performance at pressures more than 10 times higher. PMID- 19682926 TI - Adaptations of upper trapezius muscle activity during sustained contractions in women with fibromyalgia. AB - The study compared the distribution of electromyographic (EMG) signal amplitude in the upper trapezius muscle in 10 women with fibromyalgia and in 10 healthy women before and after experimentally-induced muscle pain. Surface EMG signals were recorded over the right upper trapezius muscle with a 10 x 5 grid of electrodes during 90 degrees shoulder abduction sustained for 60s. The control subjects repeated the abduction task following injections of isotonic and hypertonic (painful) saline into the upper trapezius muscle. The EMG amplitude was computed for each electrode pair and provided a topographical map of the distribution of muscle activity. The pain level rated by the patients at the beginning of the sustained contraction was 5.9+/-1.5. The peak pain intensity for the control group following the injection of hypertonic saline was 6.0+/-1.6. During the sustained contractions, the EMG amplitude increased relatively more in the cranial than caudal region of the upper trapezius muscle for the control subjects (shift in the distribution of EMG amplitude: 2.3+/-1.3mm; P<0.01). The patient group showed lower average EMG amplitude than the controls during the contraction (P<0.05) and did not show different changes in EMG amplitude between different regions of the upper trapezius. A similar behavior was observed for the control group following injection of hypertonic saline. The results indicate that muscle pain prevents the adaptation of upper trapezius activity during sustained contractions as observed in non-painful conditions, which may induce overuse of similar muscle compartments with fatigue. PMID- 19682925 TI - Shifting unoccupied spectral space in mass spectrum of peptide fragment ions. AB - Ions near the high-end border of a mass defect distribution plot for native peptide fragment ions have potential as signature markers that are based on mass to-charge ratio determination. The specificity of these marker ions, including phosphoryl ions, can be improved by removing interfering isobaric ions from the border region on the distribution plot. These interfering ions are rich in Asp and Glu content. The masses of amino acid residues and peptides are rescaled from the IUPAC scale (12C = 12 u as the mass reference) to the averagine scale (averagine mass = 111 u* as the mass reference with zero mass defect; u*: the mass unit on the averagine scale), using a scaling factor of 0.999493894. It is theoretically predicted that esterification of Asp and Glu side-chain carboxylates with n-butanol can achieve a sufficient retreat of the high-end border on a mass defect distribution plot based on the use of mass spectrometers with better-than-medium resolution. Theoretical calculations and laboratory experiments are performed to examine effects of various esterifications on the averagine-scale mass defect distribution of peptide fragment ions and on the specificity of two positive phosphoryl ions: the phosphotyrosine immonium ion and a cyclophosphoramidate ion. PMID- 19682927 TI - Psychogenic non-epileptic seizures--definition, etiology, treatment and prognostic issues: a critical review. AB - In this review we systematically assess our currently available knowledge about psychogenic non-epileptic seizures (PNES) with an emphasis on the psychological mechanisms that underlie PNES, possibilities for psychological treatment as well as prognosis. Relevant studies were identified by searching the electronic databases. Case reports were not considered. 93 papers were identified; 65 of which were studies. An open non-randomized design, comparing patients with PNES to patients with epilepsy is the dominant design. A working definition for PNES is proposed. With respect to psychological etiology, a heterogeneous set of factors have been identified. Not all factors have a similar impact, though. On the basis of this review we propose a model with several factors that may interact in both the development and prolongation of PNES. These factors involve psychological etiology, vulnerability, shaping, as well as triggering and prolongation factors. A necessary first step of intervention in patients with PNES seems to be explaining the diagnosis with care. Although the evidence for the efficacy of additional treatment strategies is limited, variants of cognitive (behavioural) therapy showed to be the preferred type of treatment for most patients. The exact choice of treatment should be based on individual differences in the underlying factors. Outcome can be measured in terms of seizure occurrence (frequency, severity), but other measures might be of greater importance for the patient. Prognosis is unclear but studies consistently report that 1/3rd to 1/4th of the patients become chronic. PMID- 19682928 TI - Interleukin-17-producing gammadelta T cells selectively expand in response to pathogen products and environmental signals. AB - Gammadelta T cells are an innate source of interleukin-17 (IL-17), preceding the development of the adaptive T helper 17 (Th17) cell response. Here we show that IL-17-producing T cell receptor gammadelta (TCRgammadelta) T cells share characteristic features with Th17 cells, such as expression of chemokine receptor 6 (CCR6), retinoid orphan receptor (RORgammat), aryl hydrocarbon receptor (AhR), and IL-23 receptor. AhR expression in gammadelta T cells was essential for the production of IL-22 but not for optimal IL-17 production. In contrast to Th17 cells, CCR6(+)IL-17-producing gammadelta T cells, but not other gammadelta T cells, express Toll-like receptors TLR1 and TLR2, as well as dectin-1, but not TLR4 and could directly interact with certain pathogens. This process was amplified by IL-23 and resulted in expansion, increased IL-17 production, and recruitment of neutrophils. Thus, innate receptor expression linked with IL-17 production characterizes TCRgammadelta T cells as an efficient first line of defense that can orchestrate an inflammatory response to pathogen-derived as well as environmental signals long before Th17 cells have sensed bacterial invasion. PMID- 19682929 TI - Interleukin-1 and IL-23 induce innate IL-17 production from gammadelta T cells, amplifying Th17 responses and autoimmunity. AB - Th17 cells, CD4(+) T cells that secrete interleukin-17 (IL-17), are pathogenic in autoimmune diseases and their development and expansion is driven by the cytokines IL-6, TGF-beta, IL-21, IL-1, and IL-23. However, there are also innate sources of IL-17. Here, we show that gammadelta T cells express IL-23R and the transcription factor RORgammat and produce IL-17, IL-21, and IL-22 in response to IL-1beta and IL-23, without T cell receptor engagement. IL-17-producing gammadelta T cells were found at high frequency in the brain of mice with experimental autoimmune encephalomyelitis (EAE). gammadelta T cells activated by IL-1beta and IL-23 promoted IL-17 production by CD4(+) T cells and increased susceptibility to EAE, suggesting that gammadelta T cells act in an amplification loop for IL-17 production by Th17 cells. Our findings demonstrate that gammadelta T cells activated by IL-1beta and IL-23 are an important source of innate IL-17 and IL-21 and provide an alternative mechanism whereby IL-1 and IL-23 may mediate autoimmune inflammation. PMID- 19682931 TI - Abiotrophia/Granulicatella tubo-ovarian abscess in an adolescent virginal female. AB - Tubo-ovarian abscess (TOA) is a common acute complication of pelvic inflammatory disease (PID). It can also develop as a complication of pelvic or abdominal surgery, malignancy, and intra-abdominal processes such as appendicitis. In premenopausal women, PID is the most common cause of tubo-ovarian abscess. We report a case of tubo-ovarian abscess in a virginal adolescent female with no past surgical history and no known history of appendicitis, inflammatory bowel disease, or cancer. Cultures of the tubo-ovarian abscess drainage grew Abiotrophia/Granulicatella species. This case supports including TOA in the broad differential diagnosis for abdominal pain with fever in adolescent females regardless of sexual history. PMID- 19682930 TI - Loss of the LAT adaptor converts antigen-responsive T cells into pathogenic effectors that function independently of the T cell receptor. AB - Despite compromised T cell antigen receptor (TCR) signaling, mice in which tyrosine 136 of the adaptor linker for activation of T cells (LAT) was constitutively mutated (Lat(Y136F) mice) accumulate CD4(+) T cells that trigger autoimmunity and inflammation. Here we show that equipping postthymic CD4(+) T cells with LATY136F molecules or rendering them deficient in LAT molecules triggers a lymphoproliferative disorder dependent on prior TCR engagement. Therefore, such disorders required neither faulty thymic T cell maturation nor LATY136F molecules. Unexpectedly, in CD4(+) T cells recently deprived of LAT, the proximal triggering module of the TCR induced a spectrum of protein tyrosine phosphorylation that largely overlapped the one observed in the presence of LAT. The fact that such LAT-independent signals result in lymphoproliferative disorders with excessive cytokine production demonstrates that LAT constitutes a key negative regulator of the triggering module and of the LAT-independent branches of the TCR signaling cassette. PMID- 19682932 TI - Expression of bovine granulocyte chemotactic protein-2 (GCP-2) in neutrophils and a mammary epithelial cell line (MAC-T) in response to various bacterial cell wall components. AB - CXC chemokines are potential attractants for polymorphonuclear leucocytes (PMNs) and play an important role in resistance to infectious diseases, such as bovine mastitis. In this study, a bovine mammary epithelial cell line (MAC-T) and blood PMNs were stimulated with bacterial cell wall components of gram negative and gram positive bacteria, including lipopolysaccharide (LPS), lipoteichoic acid (LTA) and peptidoglycan (PGN). The expression of two CXC chemokines, interleukin (IL)-8 and granulocyte chemotactic protein (GCP)-2 was analysed by real-time PCR. High concentrations (1 or 10 MUg/mL) of LPS and LTA, but not PGN, significantly increased the expression of GCP-2 and IL-8 in both MAC-T and PMNs. Biopsies from mammary glands of cattle with clinical Escherichia coli mastitis also had increased expression of GCP-2. Using an in vitro transepithelial migration assay, recombinant human GCP-2 (rhGCP-2) showed weak chemoattractant effects on bovine blood PMNs. It was concluded that PMNs and MAC-T cells can express GCP-2 in response to certain bacterial cell components during the course of mastitis. PMID- 19682935 TI - Novel dietary strategies can improve the outcome of weight loss programmes in obese client-owned cats. AB - A randomised, single-blinded, positively controlled, field trial for weight loss in obese client-owned cats was undertaken to look at novel diets and dietary strategies that could improve owner compliance and, therefore, success of feline weight loss programmes. Three dietary strategies were evaluated: strategy A used a novel dry high fibre ration; strategy B used ready-prepared portions of dry and moist food; strategy C used an existing commercial dry high fibre ration fed with a measuring cup. Cats were assessed at weeks 4, 12 and 20, and adjustments to the energy allocation made if required. Mean weight loss at 20 weeks (A: 11.0+/-1.3%, B: 10.9+/-1.2%, C: 11.9+/-1.7%) and mean energy allocation (A: 31.0, B: 28.5 and C: 32.2 kcal/kg of target body weight/day) were similar amongst strategies. However, owners' subjective hunger score was significantly (P<0.001) higher with strategy C than with strategies A and B. Further, owner satisfaction was lowest with strategy C, and more owners also regarded food allowance as insufficient with this strategy. Novel diets and feeding strategies may improve outcome in feline weight loss programmes. PMID- 19682936 TI - Disseminated cryptococcosis resulting in miscarriage in a woman without other immunocompromise: a case report. AB - We present an unusual case of disseminated cryptococcosis involving the lungs, placenta, and gall bladder in an apparently immunocompetent pregnant woman. The infection resulted in spontaneous abortion. The patient's condition only improved after cholecystectomy and several weeks of antifungal therapy. An in-depth evaluation revealed no central nervous system involvement or immunocompromising condition other than pregnancy. PMID- 19682934 TI - Linking cell cycle to histone modifications: SBF and H2B monoubiquitination machinery and cell-cycle regulation of H3K79 dimethylation. AB - To identify regulators involved in determining the differential pattern of H3K79 methylation by Dot1, we screened the entire yeast gene deletion collection by GPS for genes required for normal levels of H3K79 di- but not trimethylation. We identified the cell cycle-regulated SBF protein complex required for H3K79 dimethylation. We also found that H3K79 di- and trimethylation are mutually exclusive, with M/G1 cell cycle-regulated genes significantly enriched for H3K79 dimethylation. Since H3K79 trimethylation requires prior monoubiquitination of H2B, we performed genome-wide profiling of H2BK123 monoubiquitination and showed that H2BK123 monoubiquitination is not detected on cell cycle-regulated genes and sites containing H3K79me2, but is found on H3K79me3-containing regions. A screen for genes responsible for the establishment/removal of H3K79 dimethylation resulted in identification of NRM1 and WHI3, both of which impact the transcription by the SBF and MBF protein complexes, further linking the regulation of methylation status of H3K79 to the cell cycle. PMID- 19682937 TI - [Indications and technical procedures of mastectomy with immediate breast reconstruction]. AB - Achieving a mastectomy occurs in one third of the acts of breast cancer surgery. In the present context, the increase in the diagnosis of early breast cancer, including widespread or multifocal forms, the possibility of a reconstruction at the same time is discussed. It will be proposed in a specific context, taking into account various factors: oncological (characteristics of the tumor, adjuvant treatments), clinical (smoking, comorbidity) and psychological (choice of the patient). The technique of mastectomy will be adapted to the indication and choice of reconstruction procedure. Including the preservation of the nipple areola complex, or the skin envelope will be possible for some guidance. The reconstruction is possible with the use of breast implants, or musculocutaneous flaps or by the combination of both. Some of these techniques are associated with significant morbidity, and appropriate selection of procedure to the patient will guarantee a lower risk of complications. PMID- 19682938 TI - [Let's put an end to systematic axillary lymph node dissection in breast tumors more than 2cm!]. PMID- 19682939 TI - [Heterotopic ovarian hydatidiform mole. A case report]. AB - We report a case of ruptured ovarian molar pregnancy, diagnosed in the context of haemorragic shock 19 days following evacuation of an intrauterine hydatiform mole. To our knowledge, this is the first reported case of heterotopic molar pregnancy. PMID- 19682940 TI - [Maternal and neonatal consequences induced by the French recommendations for episiotomy practice. Monocentric study about 5409 vaginal deliveries]. AB - OBJECTIVES: Recommendations for clinical practice (RPC) edited by the College of French gynecologists and obstetricians (CNGOF) claim for a more restrictive use of episiotomy. The aims of this study were to assess the impact of these recommendations on episiotomy practice and to evaluate maternal and neonatal outcomes of a more restrictive approach. PATIENTS AND METHODS: We compared in a retrospective analysis episiotomy practice, maternal and neonatal consequences of a restrictive episiotomy policy between 2004 and 2006 (before and after recommendations publication) in a level III maternity unit. Identification of risks factors for episiotomy practice in our population was based on a multivariate analysis. RESULTS: Two thousand and five hundred and 2909 patients who delivered vaginally respectively in 2004 and 2006 were included. In 2006, fewer episiotomies were performed (43.48% vs 32.32%, p<0.0001) and more grade I and II perineal tears occurred (27.56% vs 36.61%, p=0.0001) whereas the anal sphincter tear rate remained constant (0.48% vs 0.69%, p=0.376). Neonatal condition assessed by the Apgar score was also stable. In multivariate analysis, risk factors for episiotomy already described in the literature were significant in our study and year 2006 was associated with less use of episiotomy in comparison with year 2004 (OR: 0.499; IC: 0.44-0.57; p<0.0001). DISCUSSION AND CONCLUSION: In our institution, French guidelines were followed by a reduction in the use of episiotomy practice without increasing the risk for severe perineal tear or neonatal distress. PMID- 19682941 TI - [Slow freezing and vitrification of human mature and immature oocytes]. AB - Performance and security questions in human oocyte cryopreservation have been taking researchers for about two decades. Oocytes are usually frozen at metaphase II. Immature oocytes cryopreservation is still a research alternative. Two techniques are currently available for oocyte cryopreservation: slow freezing and vitrification. Experimental data suggest that vitrification has less impact on oocyte physiology than classical slow freezing. After slow freezing of mature oocytes, survival and fertilization rates reach 70 to 80% whereas cleavage rates are around 90%, leading to five implantations and 1.2 births per 100 thawed oocytes. After vitrification of mature oocytes, survival and cleavage rates reach 90% leading to 11 implantations and 1.8 births per 100 thawed oocytes. The obstetrical and neonatal prognosis of these pregnancies is reassuring. No increased risk of congenital anomalies has been observed. However, further evaluation is needed to guarantee the safety of cryopreservation procedures. Immature oocyte cryopreservation is not currently perfected but some indications appear of great interest. PMID- 19682942 TI - [The Bcl-2 family pathway in gametes and preimplantation embryos]. AB - Apoptosis, a form of cell death by self-destruction, has been reported in gametes and preimplantation embryos both in vitro and in vivo. Recent evidence suggests that cell death processes, whose control deserves to be elucidated, can impact embryo developmental competence. Moreover, quality of the gametes (particularly of the oocytes) is relevant not only for their survival rates but exert an influence during the early stages of embryo development. Thus, the investigation of apoptosis-related genes and mechanisms in early embryos is crucial. BCL-2 family proteins, through balanced interactions between pro- and anti-death members, play a pivotal role in controlling cell life and death. In this article, we review the literature concerning the expression of Bcl-2 family members in gametes and early embryos. Research results indicate that the various Bcl-2 subfamilies (pro- and anti-apoptotic "multidomain" family members and "BH3-only" death factors) exhibit a dynamic expression pattern during male and female gamete differentiation and early embryo development. While pro-apoptotic Bax protein plays a critical role in germ cell and early embryo degeneration, the relative importance of the prosurvival (Bcl-2, Bcl-xL, Bcl-w, Mcl-1) and "BH3-only" (Bim, Bad, Bik) members is not clear. Although information on expression patterns of Bcl-2 family transcripts and proteins is necessary, other elements such as transcriptional control (by environmental stimuli), subcellular localization and post-translational modifications should also be taken into account. Aside from basic research, a better understanding of apoptosis-related proteins and mechanisms involved in gamete and embryo viability at the molecular level may provide new guides for diagnosis and therapeutic strategies. PMID- 19682943 TI - Risks of intracranial hemorrhage in patients with Parkinson's disease receiving deep brain stimulation and ablation. AB - OBJECTIVES: This study analyzed risk factors for hemorrhage in a large series of deep brain stimulation (DBS) and ablation procedures in patients with advanced Parkinson's disease (PD). METHODS: Six hundred and forty four subjects with advanced PD treated with DBS or ablation procedures between March 1999 and December 2007 were enrolled in the study. Procedures were performed by the same surgeon, and included DBS in 126 patients, ablation in 507 patients and DBS after prior unilateral ablation procedures in 11 patients. Of 796 target procedures, 207 were DBS including 202 subthalamic nucleus (STN) targets, 3 ventralis intermedius nucleus (Vim) targets and 2 globus pallidus internus (GPi) targets, and the others were 589 ablation procedures including 474 GPi targets and 115 Vim targets. Postoperative CT or MRI was performed in all patients within 24 h of lead implantation or ablation treatment. Statistical correlation analysis of risk factors for intracranial hemorrhage (ICH) was performed by stepwise logistic regression. Explanatory variables were patient age, sex, blood pressure, anatomical targets, the number of microelectrode recording (MER) penetrations and surgical modality. RESULTS: Postoperative symptomatic ICH occurred in 10 cases (8 pallidotomy and 2 thalamotomy) and asymptomatic ICH in 14 cases (9 pallidotomy, 4 thalamotomy and 1 DBS). Hypertension and surgical modality were significant factors contributing to hemorrhage (both P < 0.05). The likelihood of hemorrhage in hypertensive patients was 2.5 times that in normotensive patients. The risk of hemorrhage during ablation was 5.4 times that in DBS. The number of MER trajectories did not significantly correlate with ICH occurrence (P = 0.07). No statistically significant difference was found in age, sex and anatomical targets. CONCLUSION: This study demonstrated that hypertension is a risk factor for ICH in PD patients. DBS is generally a safe surgical modality as compared with ablation. Increasing microelectrode trajectories seemed to increase the risk of ICH, but no statistically significant difference was found (P = 0.07). PMID- 19682944 TI - Vascular risk factors do not contribute to motor and cognitive impairment in Parkinson's disease. PMID- 19682945 TI - Constrained non-rigid registration for use in image-guided adaptive radiotherapy. AB - A constrained non-rigid registration (CNRR) algorithm for use in prostate image guided adaptive radiotherapy is presented in a coherent mathematical framework. The registration algorithm is based on a global rigid transformation combined with a series of local injective non-rigid multi-resolution cubic B-spline Free Form Deformation (FFD) transformations. The control points of the FFD are used to non-rigidly constrain the transformation to the prostate, rectum, and bladder. As well, the control points are used to rigidly constrain the transformation to the estimated position of the pelvis, left femur, and right femur. The algorithm was tested with both 3D conformal radiotherapy (3DCRT) and intensity-modulated radiotherapy (IMRT) dose plan data sets. The 3DCRT dose plan set consisted of 10 fan-beam CT (FBCT) treatment-day images acquired from four different patients. The IMRT dose plan set consisted of 32 cone-beam CT (CBCT) treatment-day images acquired from 4 different patients. The CNRR was tested with different combinations of anatomical constraints and each test significantly outperformed both rigid and non-rigid registration at aligning constrained bones and critical organs. The CNRR results were used to adapt the dose plans to account for patient positioning errors as well as inter-day bone motion and intrinsic organ deformation. Each adapted dose plan improved performance by lowering radiation distribution to the rectum and bladder while increasing or maintaining radiation distribution to the prostate. PMID- 19682946 TI - Fluorescent biosensors for real-time tracking of post-translational modification dynamics. AB - Dynamic post-translational modifications (PTMs) regulate and diversify protein properties and cellular behaviors. Real-time monitoring of these modifications has been made possible with biosensors based on fluorescent proteins (FPs) and fluorescence resonance energy transfer (FRET), which can provide spatiotemporal information of PTMs with little perturbation to the cellular environment. In this review, we highlight available fluorescent biosensors applicable to detect PTMs in living cells and how they have shed light on biological questions that have been difficult to address otherwise. In addition, we also provide discussions about various engineering strategies for overcoming potential challenges associated with the development and application of such biosensors. PMID- 19682947 TI - Redox, thermodynamic and spectroscopic of some transition metal complexes containing heterocyclic Schiff base ligands. AB - Complexes of two series of Schiff base ligands, H(2)L(a) and H(2)L(b) derived from the reaction of 2,6-diacetyl pyridine with semicarbazide, H(2)L(a) and thiosemicarbazide, H(2)L(b), with the metal ions, Co(II), Ni(II), Cu(II), VO(IV) and UO(2)(VI) have been prepared. The ligands are characterized by elemental analysis, IR, UV-vis and (1)H NMR. The structures of the complexes are investigated with the IR, UV-vis, X-band ESR spectra, (1)H NMR and thermal gravimetric analysis as well as conductivity and magnetic moment measurements. The IR-spectra reveal the presence of variable modes of chelation for the investigated ligands. A variety of binuclear or mononuclear complexes were obtained with the two ligands in tri-, tetra or pentadentate forms. The bonding sites are the pyridine nitrogen, two azomethine nitrogen atoms and ketonic oxygen in case of H(2)L(a) or sulphur atoms in case of H(2)L(b). The Coats-Redfern equation has been used to calculate the kinetic and thermodynamic parameters for the different thermal decomposition steps of some complexes. Cyclic voltammograms of Co(II) and Ni(II) show quasi-reversible peaks. The redox properties and the nature of the electro-active species of the complexes have been characterized. PMID- 19682948 TI - HIV-1 pol phylogenetic diversity and antiretroviral resistance mutations in treatment naive patients from Central West Brazil. AB - BACKGROUND: Primary resistance represents a challenge for the control of HIV-1 because it can reduce the efficacy of first line antiretroviral therapy and may impact clinical outcomes. OBJECTIVES: To describe the prevalence of primary HIV-1 drug resistance and subtypes circulating in Central West Brazil. STUDY DESIGN: 103 antiretroviral naive patients were recruited in Goiania city, Central West Brazil during 2007-2008. Protease and partial reverse transcriptase regions were retrotranscribed from plasma HIV-1 RNA and 97 were sequenced after direct nested PCR. HIV-1 subtype was assigned by phylogenetic analysis. Primary drug resistance was analyzed by the Calibrated Population Resistance (CPR) tool using Stanford Surveillance Drug Resistance Mutation (SDRM) and International AIDS Society-USA (IAS-USA) major mutation list. RESULTS: Primary drug resistance mutations ranged from 8% (IAS) to 10% (SDRM). High level resistance to at least one antiretroviral drug was observed. T215D/S revertant mutations were identified in 4/97 patients. HIV-1 subtype B represented 82.5%, subtype F1 6.2%, subtype C 3.1%, B/F1 7.2% and one sample was a F1/C/B mosaic. HIV-1 subtype C sequences formed a monophyletic cluster with other Brazilian subtype C sequences. CONCLUSIONS: Our HIV-1 pol sequences from Central West region include the first inland HIV-1 subtype C sequences and help compose the molecular epidemiology map of HIV-1 in Brazil. This data also show that a significant proportion of the naive patients presented important drug resistance mutations. Therefore naive patients from this setting may benefit from pre-treatment genotypic testing to optimize the choice of antiretroviral drugs and to help control HIV-1 transmission. PMID- 19682949 TI - Simplified cerebral protection using unilateral antegrade cerebral perfusion and moderate hypothermic circulatory arrest. AB - BACKGROUND: Antegrade cerebral perfusion is one of the most reliable methods of organ protection during hypothermic circulatory arrest for aortic arch surgery. We used a simplified antegrade cerebral perfusion technique with low mortality and morbidity. METHODS: Between January 2005 and August 2008, 21 patients underwent aortic arch surgery with unilateral antegrade selective cerebral perfusion through the brachiocephalic artery and moderate hypothermic circulatory arrest. The mean age for patients was 58.0+/-11.1 (27-82) years. Cardiopulmonary bypass was commenced and the ascending aorta was cross-clamped. Patients were cooled to 22-28 degrees C, whilst the proximal anastomosis was performed. The brachiochephalic artery was cannulated using a balloon tipped 15Fr catheter used for retrograde cardioplegia. Antegrade cerebral perfusion was established at the rate of 10 ml//kg/min. The perfusion pressure was controlled between 50 and 70 mm Hg whilst the distal anastomosis was completed. RESULTS: There were no operative deaths and no permanent neurological deficits. Four patients had temporary confusion. Mean antegrade cerebral perfusion time was 21.6+/-8.0 (12-48)min. Eight out of 20 patients had circulatory arrest at 28 degrees C and their mean circulatory arrest time was 22.8+/-4.7 (16-32)min. DISCUSSION: The mortality and neurological outcomes of aortic surgery using unilateral antegrade cerebral perfusion with moderate hypothermic circulatory arrest produced satisfactory results. Bilateral cannulation and deep hypothermia appear to be unnecessary in most cases. The coagulopathy from deep hypothermia is thereby avoided. PMID- 19682950 TI - Scientific knowledge and fundamental practices of platelet transfusion. PMID- 19682951 TI - Selecting effective siRNA target sequences by using Bayes' theorem. AB - Short interfering RNA (siRNA) has been widely used for studying gene functions in mammalian cells but varies markedly in its gene silencing efficacy. Although many design rules/guidelines for effective siRNAs based on various criteria have been reported recently, there are few consistencies among them. This makes it difficult to select effective siRNA sequences in mammalian genes. Another shortcoming of most previously reported methods is that they cannot estimate the probability that a candidate sequence will silence the target gene. The analytical prediction method proposed in the present study uses Bayes' theorem to select effective siRNA target sequences from many possible candidate sequences. It is quite different from the previous score-based siRNA design techniques and can predict the probability that a candidate siRNA sequence will be effective. The results of evaluating it by applying it to recently reported effective and ineffective siRNA sequences for various genes indicate that it would be useful for many other genes. It should therefore be useful for selecting siRNA sequences effective for mammalian genes. PMID- 19682952 TI - Retroperitoneoscopic para-aortic lymph node sampling in bladder rhabdomyosarcoma. AB - Determining lymph node involvement is an important step in the pre-treatment evaluation of non-metastatic rhabdomyosarcoma. We describe retroperitoneoscopy for para-aortic lymph node biopsy in a 4-year-old boy with embryonal rhabdomyosarcoma of the bladder with pelvic and para-aortic lymph node enlargement on magnetic resonance imaging. This technique affords access to the para-aortic region with minimal dissection, permitting quick recovery and early commencement of chemotherapy. PMID- 19682953 TI - Health behaviors and conditions of persons with epilepsy: a bivariate analysis of 2006 BRFSS data. AB - Using amalgamated Behavioral Risk Factor Surveillance System (BRFSS) data from nine states in 2006, this study examined the differences in demographics, health conditions, health behaviors, and quality of life among persons with active epilepsy (PWAE) and persons with a lifetime prevalence of epilepsy (PWLE), compared with persons without epilepsy (PWOE). Frequencies for PWAE, PWLE, and PWOE by selected health behaviors, health conditions, quality of life, and demographic variables were produced. Differences between persons with and without epilepsy were analyzed. Results indicated smoking and unemployment rates were significantly higher among PWLE/PWAE than among PWOE. PWLE/PWAE were also less likely than PWOE to report advanced educational attainment, and more likely to report suffering physical, mental, or emotional limitations that affected their activities in some way. Hispanics were less likely to report a lifetime or active prevalence of epilepsy than people without epilepsy. PWLE/PWAE report several differences in demographics, health conditions, health behaviors, and quality of life compared to people without epilepsy. PMID- 19682954 TI - Memory performance is related to language dominance as determined by the intracarotid amobarbital procedure. AB - OBJECTIVE: The goal of this study was to explore the relationship between language and memory lateralization in patients with epilepsy undergoing the intracarotid amobarbital procedure. METHODS: In 386 patients, language lateralization and memory lateralization as determined by laterality index (LI) were correlated with each other. RESULTS: Language lateralization and memory lateralization were positively correlated (r=0.34, P<0.01). Correlations differed depending on the presence and type of lesion (chi(2)=7.98, P<0.05). LIs correlated significantly higher (z=2.82, P<0.05) in patients with cortical dysplasia (n=41, r=0.61, P<0.01) compared with the group without lesions (n=90, r=0.16, P>0.05), with patients with hippocampal sclerosis falling between these two groups. Both memory (P<0.01) and language (P<0.01) LIs were higher in right- compared with left-sided lesions. CONCLUSION: Correlation of language and memory is more pronounced in patients with structural lesions as compared with patients without lesions on MRI. PMID- 19682955 TI - Anticonvulsant effects of the selective melatonin receptor agonist ramelteon. AB - OBJECTIVE: The endogenous hormone melatonin has previously been shown to exert anticonvulsant effects in a variety of experimental models. Accordingly, we asked whether ramelteon, a synthetic and selective melatonin receptor agonist, might also possess anticonvulsant and/or antiepileptogenic properties. METHODS: The effects of ramelteon (30 or 100 mg/kg intraperitoneally twice daily for 5 days) were evaluated in two animal models of epilepsy. In the rat rapid kindling model, baseline hippocampal afterdischarge properties, kindling progression, and hippocampal excitability in kindled animals were measured. Anti-ictogenic efficacy was assessed after acute administration in untreated kindled rats. In the spontaneously epileptic Kcna1-null mouse model, we determined seizure frequency and periodicity using continuous video/EEG monitoring over 72 hours. Further, circadian rest-activity rhythms in ramelteon-treated animals were studied with actigraphy. RESULTS: In kindled animals, ramelteon reversed kindling induced hippocampal excitability; however, it did not modify baseline afterdischarge properties, the progression and establishment of the kindled state in the rapid kindling model. However, in Kcna1-null mice, ramelteon (200 mg/kg/day) significantly attenuated seizure periodicity and frequency and improved circadian rest-activity rhythms compared with control animals. CONCLUSIONS: The selective melatonin receptor agonist ramelteon possesses anticonvulsant properties in a chronic epilepsy model. Our findings provide further support for melatonin receptors being potential novel targets for anticonvulsant drug development. PMID- 19682956 TI - Structure-function relationship of sensory endings in the gut and bladder. AB - Visceral afferents play a key role in neural circuits underlying the physiological function of visceral organs. They are responsible for the detection and transmission of a variety of visceral sensations (e.g. satiety, urge, discomfort and pain) from the viscera to the central nervous system. A comprehensive account of the different functional types of visceral sensory neurons would be invaluable in understanding how sensory dysfunction occurs and how it might be diagnosed and treated. Our aim was to explore the morphology of different nerve endings of visceral afferents within the gastrointestinal tract and urinary bladder and how the morphology of these nerve endings may relate to their functional properties. Morphological studies of mechanosensitive endings of visceral afferents to the gut and bladder correlated with physiological recordings have added a new dimension to our ability to distinguish different functional classes of visceral afferents. PMID- 19682958 TI - Flow injection determination of ketotifen fumarate using PVC membrane selective electrodes. AB - In this study a PVC membrane electrode for determination of ketotifen fumarate is reported, where ketotifen tetraphenylborate (Keto-TPB) was used as ion exchanger. The electrode has linear range of 5.6x10(-6)-1.0x10(-2) and 1.0x10(-5)-1.0x10(-2) mol/L, with detection limits 2.37x10(-6)and 4.60x10(-6) mol/L in batch and flow injection analysis (FIA), respectively. The electrodes show a Nernstian slope value (58.40 and 61.50 mV/decade in batch and FIA, respectively), and the response time is very short (10(8)) is impeded by the limited transformation efficiency of Escherichia coli. Therefore, it is currently difficult to create a mutant library containing amino acid substitutions in more than seven residues. To overcome this problem, here we have used two different TNF mutant libraries, each containing random substitutions at six selected amino acid residues, and utilized a gene shuffling method to construct a randomized mutant library containing substitutions at 12 different amino acid residues of TNF. Consequently, using this library, we identified TNF mutants with greater receptor-selectivity and enhanced receptor-specific bioactivity than the existing mutants. PMID- 19682975 TI - Detection of conformationally changed MBP using intramolecular FRET. AB - The principal objective of this study was to explore protein conformational changes using fluorescence resonance energy transfer (FRET) technology. Maltose binding protein (MBP) was adopted as a target model, due to its well characterized structure and ligand specificity. To the best of our knowledge, this is the first report to provide information regarding the biological distance between the two lobes of MBP upon maltose binding. For the FRET pair, ECFP and EYFP were used as the donor and the acceptor, and were linked genetically to the C-terminal and N-terminal regions of MBP (ECFP:MBP:EYFP), respectively. After the FRET reaction, maltose-treated MBP was shown to exhibit a considerable energy transfer (FRET efficiency (E)= approximately 0.11, Distance (D)= approximately 6.93 nm) at the ensemble level, which was regarded as reflective of the increase in donor quenching and the upshift in acceptor emission intensity, thereby suggesting that the donor and the acceptor had been brought close together as the result of structural alterations in MBP. However, upon glucose treatment, no FRET phenomenon was detected, thereby implying the specificity of interaction between MBP and maltose. The in vitro FRET results were also confirmed via the acceptor photobleaching method. Therefore, our data showed that maltose-stimulated conformational changes of MBP could be measured by FRET, thereby providing biological information, including the FRET efficiency and the intramolecular distance. PMID- 19682976 TI - Structure of the Enterococcus faecalis EIIA(gnt) PTS component. AB - In Eubacteria, the utilization of a number of extracellular carbohydrates is mediated by sugar specific phosphoenolepyruvate (PEP) dependent sugar phosphotransferase systems (PTSs), which simultaneously import und phosphorylate their target sugars. Here, we report the crystal structure of the EIIA(gnt) component of the so far little investigated Enterococcus faecalis gluconate specific PTS. The crystal structure shows a tightly interacting dimer of EIIA(gnt) which is structurally similar to the related EIIA(man) from Escherichia coli. Homology modeling of E. faecalis HPr, EIIB(man) and their complexes with EIIA(man) suggests that despite moderate sequence identity between EIIA(man) and EIIA(gnt), the active sites closely match the situation observed in the E. coli system with His-9 of EIIA(gnt) being the likely phosphoryl group carrier. We therefore propose that the phosphoryl transfer reactions involving EIIA(gnt) proceed according to a mechanism analog to the one described for E. coli EIIA(man). PMID- 19682977 TI - Cardiac action potential duration and calcium regulation in males and females. AB - Adult women have longer QT intervals compared with men of a similar age, indicating differences in the speed of repolarisation of the ventricles. We investigate the influences of gender on ventricular electrophysiology and intracellular Ca(2+) regulation of the guinea pig heart. Comparing sexually mature animals, females exhibited a significantly longer APD. Peak L-type Ca(2+) current (I(CaL)) was larger in females and when this current was inhibited with nifedipine the gender differences in APD were removed. APD differences also disappeared when the SR was depleted of Ca(2+). Inactivation of I(CaL) during a clamp step is faster in females but slower during an action potential and SR Ca(2+) content is larger. We suggest that gender differences in APD result from variation in the kinetics of I(CaL) stemming from alterations to Ca(2+) release. PMID- 19682978 TI - The conjugated linoleic acid isomer trans-9,trans-11 is a dietary occurring agonist of liver X receptor alpha. AB - Conjugated linoleic acid (CLA) isomers are dietary fatty acids that modulate gene expression in many cell types. We have previously reported that specifically trans-9,trans-11 (t9,t11)-CLA induces expression of genes involved in lipid metabolism of human macrophages. To elucidate the molecular mechanism underlying this transcriptional activation, we asked whether t9,t11-CLA affects activity of liver X receptor (LXR) alpha, a major regulator of macrophage lipid metabolism. Here we show that t9,t11-CLA is a regulator of LXRalpha. We further demonstrate that the CLA isomer induces expression of direct LXRalpha target genes in human primary macrophages. Knockdown of LXRalpha with RNA interference in THP-1 cells inhibited t9,t11-CLA mediated activation of LXRalpha including its target genes. To evaluate the effective concentration range of t9,t11-CLA, human primary macrophages were treated with various doses of CLA and well known natural and synthetic LXR agonists and mRNA expression of ABCA1 and ABCG1 was analyzed. Incubation of human macrophages with 10 microM t9,t11-CLA led to a significant modulation of ABCA1 and ABCG1 transcription and caused enhanced cholesterol efflux to high density lipoproteins and apolipoprotein AI. In summary, these data show that t9,t11-CLA is an agonist of LXRalpha in human macrophages and that its effects on macrophage lipid metabolism can be attributed to transcriptional regulations associated with this nuclear receptor. PMID- 19682979 TI - Alpha-helical transmembrane peptides: a "divide and conquer" approach to membrane proteins. AB - Alpha-helical membrane proteins fulfill many vital roles in all living cells and constitute the majority of drug targets. However, their relevance is in no way paralleled by our current understanding of their structures and functions. This is because membrane proteins present a number of experimental obstacles that are difficult to surmount by classical methods developed for water-soluble proteins. Moreover, membrane proteins are not only challenging on their very own but, when embedded in a biological membrane, also reside in an outstandingly complex milieu. These difficulties have fostered a "divide and conquer" approach, in which a membrane protein is dissected into shorter and easier-to-handle transmembrane (TM) peptides. Under suitable conditions, such peptides fold independently and retain many of the properties displayed in the context of the full-length parent protein. This contribution reviews some of the most notable insights into alpha-helical membrane proteins gleaned from experiments on protein derived TM peptides. We recapitulate some peculiar properties of lipid bilayers that render them such a complex and unique environment and discuss generic features pertaining to hydrophobic peptides derived from alpha-helical membrane proteins. The main part of the review is devoted to a critical discussion of particularly interesting examples of TM peptides studied in membrane-mimetic systems of increasing complexity: isotropic solvents, detergent micelles, lipid bilayers, and biological membranes. The unifying theme is to explore to what extent TM peptides in combination with different membrane-mimetic systems can aid in advancing our knowledge and comprehension of alpha-helical membrane proteins as well as in developing new pharmacological tools. PMID- 19682980 TI - The total synthesis of 2-O-arachidonoyl-1-O-stearoyl-sn-glycero-3-phosphocholine 1,3,1'-(13)C3 and -2,1'-(13)C2 by a novel chemoenzymatic method. AB - 2-O-Arachidonoyl-1-O-stearoyl-sn-glycero-3-phosphocholine was synthesized with carbon-13 enrichment of the three glycerol carbons and the carbonyl of the stearoyl group. Phospholipase A(2) was utilized to give optically pure lyso-PC, and only 3% acyl migration occurred during reacylation with arachidonic acid anhydride. This phospholipid is an important biosynthetic precursor of arachidonic acid metabolites as well as the endocannabinoid 2 arachidonoylglycerol (2-AG), and is now available for NMR studies. PMID- 19682981 TI - Haptoglobin polymorphism in Mongolian population: comparison of the two genotyping methods. AB - BACKGROUND: Haptoglobin (Hp) polymorphisms have been suggested to be associated with many pathological conditions including cardiovascular diseases, infectious diseases, and type 2 diabetes. METHODS: We examined the association between HP genotypes and type 2 diabetes or anthropometric and clinical features in 946 Mongolians. HP genotypes were determined by two methods, TaqMan-based real-time PCR and conventional PCR. Hp phenotyping was also performed by polyacrylamide gel electrophoresis and peroxidase staining in a few selected samples. RESULTS: The 2 HP genotyping methods showed consistent results in 943 of 946 samples. Three samples that showed different results in the 2 PCR methods were determined to be heterozygous for the Hp Johnson allele by phenotyping. We did not find any association between HP genotypes and type 2 diabetes or the anthropometric and clinical features examined in the population studied. CONCLUSIONS: TaqMan-based HP genotyping is reliable and will be useful for high-throughput association studies. Further studies of large numbers of subjects and functional differences in Hp types are needed. PMID- 19682982 TI - Regulation of gene expression via the core promoter and the basal transcriptional machinery. AB - The RNA polymerase II core promoter is a structurally and functionally diverse transcriptional regulatory element. There are two main strategies for transcription initiation - focused and dispersed initiation. In focused initiation, transcription starts from a single nucleotide or within a cluster of several nucleotides, whereas in dispersed initiation, there are several weak transcription start sites over a broad region of about 50 to 100 nucleotides. Focused initiation is the predominant means of transcription in simpler organisms, whereas dispersed initiation is observed in approximately two-thirds of vertebrate genes. Regulated genes tend to have focused promoters, and constitutive genes typically have dispersed promoters. Hence, in vertebrates, focused promoters are used in a small but biologically important fraction of genes. The properties of focused core promoters are dependent upon the presence or absence of sequence motifs such as the TATA box and DPE. For example, Caudal, a key regulator of the homeotic gene network, preferentially activates transcription from DPE- versus TATA-dependent promoters. The basal transcription factors, which act in conjunction with the core promoter, are another important component in the regulation of gene expression. For instance, upon differentiation of myoblasts to myotubes, the cells undergo a switch from a TFIID based transcription system to a TRF3-TAF3-based system. These findings suggest that the core promoter and basal transcription factors are important yet mostly unexplored components in the regulation of gene expression. PMID- 19682983 TI - miR-196 is an essential early-stage regulator of tail regeneration, upstream of key spinal cord patterning events. AB - Salamanders have the remarkable ability to regenerate many body parts following catastrophic injuries, including a fully functional spinal cord following a tail amputation. The molecular basis for how this process is so exquisitely well regulated, assuring a faithful replication of missing structures every time, remains poorly understood. Therefore a study of microRNA expression and function during regeneration in the axolotl, Ambystoma mexicanum, was undertaken. Using microarray-based profiling, it was found that 78 highly conserved microRNAs display significant changes in expression levels during the early stages of tail regeneration, as compared to mature tissue. The role of miR-196, which was highly upregulated in the early tail blastema and spinal cord, was then further analyzed. Inhibition of miR-196 expression in this context resulted in a defect in regeneration, yielding abnormally shortened tails with spinal cord defects in formation of the terminal vesicle. A more detailed characterization of this phenotype revealed downstream components of the miR-196 pathway to include key effectors/regulators of tissue patterning within the spinal cord, including BMP4 and Pax7. As such, our dataset establishes miR-196 as an essential regulator of tail regeneration, acting upstream of key BMP4 and Pax7-based patterning events within the spinal cord. PMID- 19682986 TI - Transport of prion proteins across the blood-brain barrier. PMID- 19682984 TI - Does elevated intraocular pressure reduce retinal TRKB-mediated survival signaling in experimental glaucoma? AB - Reduced retrograde transport of neurotrophins (NT) and their receptors has been hypothesized to contribute directly to retinal ganglion cell (RGC) loss in glaucoma. However, strategies of supplementing NT and NT receptors have failed to avert ultimate RGC death in experimental glaucoma. This study examines the response of major components of the NT system and their interacting proteins in a rat glaucoma model. Unilateral chronic intraocular pressure (IOP) elevation was produced by episcleral vein injection of hypertonic saline (N = 99). Retinas were collected and grouped by extent of optic nerve injury. Quantitative reverse transcription PCR, western blot analysis and immunohistochemistry were used to determine mRNA and protein levels and protein localization. Out of three RGC specific Brn3 proteins (Brn3a, b, and c), only Brn3a was significantly downregulated at the message level to 35 +/- 4% of fellow values with the severest nerve injury. With IOP elevation, no significant alterations were found in retinal mRNA levels for BDNF, NGF, NT-4/5 or NT-3. The abundance of mature retinal BDNF protein was not significantly affected by elevated IOP, while proBDNF protein decreased linearly with increasing injury grade (r(2) = 0.50). In retinas with the severest nerve injury, TrkB and TrkC receptor mRNA levels significantly declined to 67 +/- 9% and 44 +/- 5% of fellow values, respectively. However, the levels of TRKB protein and its phosphorylated form were unchanged. Message level for p75(NTR) was linearly upregulated up to 219 +/- 26% with increasing injury (r(2) = 0.46), but no alteration was detected at protein level. The mRNA expression of p75(NTR) apoptosis adaptor proteins NADE, NRIF, and Lingo1 were significantly downregulated in retinas with the greatest nerve injury. A positive correlation was found between injury extent and message levels for Jun (r(2) = 0.23) as well as Junb (r(2) = 0.27), and RGC labeling of activated JUN protein increased. Atf3 mRNA levels demonstrated a positive linear correlation to the extent of injury (r(2) = 0.53), resulting in a nearly five-fold increase (482 +/- 76%) in eyes with the greatest nerve damage. Among downstream pro-survival signaling components, Erk5 mRNA expression was linearly upregulated (r(2) = 0.32) up to 157 +/- 15% of fellow values in retinas with the severest nerve injury (p < 0.01). A slight positive correlation was found between NF-kappaB message levels and injury extent (r(2) = 0.12). Bcl-xl mRNA levels in the most severely injured retinas were significantly reduced to 83 +/- 7% by elevated IOP exposure. Message levels for Erk1/2, Akt1-3 or Bcl2 appeared unaffected. Elevated IOP did not alter mRNA levels of pro-apoptotic Bim, Bax, or p53. This study demonstrates that elevated IOP exposure does not result in a dramatic decrease in retinal levels of either BDNF or its receptor, TrkB. It shows that the responses of NT pathways to elevated IOP are complex, particularly with regard to the role of p75(NTR) and Atf3. A better understanding of the roles of these proteins in IOP-induced injury is likely to suggest informed strategies for neuroprotection in glaucoma. PMID- 19682985 TI - Elovl4 5-bp deletion knock-in mouse model for Stargardt-like macular degeneration demonstrates accumulation of ELOVL4 and lipofuscin. AB - The mechanism underlying photoreceptor degeneration in autosomal dominant Stargardt-like macular degeneration (STGD3) due to mutations in the elongation of very long chain fatty acids-4 (ELOVL4) gene is not fully understood. To evaluate the pathological events associated with STGD3, we used a mouse model that mimics the human STGD3 phenotype and studied the progression of retinal degeneration. Morphological changes in the retina of Elovl4 5-bp deletion knock-in mice (E_mut(+/-)) were evaluated at 22 months of age. The localization of ELOVL4, and the expression pattern of inner retinal tissue marker proteins, and ubiquitin were determined by immunofluorescence labeling of retinal sections. Levels of the retinal pigment epithelium (RPE) lipofuscin fluorophores were measured by quantitative HPLC. Morphological evaluation of the retina revealed an accumulation of RPE debris in the subretinal space. A significant increase in the amount of ELOVL4 was observed in the outer plexiform layer in E_mut(+/-) mice compared to controls. Apart from the accumulation of ELOVL4, E_mut(+/-) mice also exhibited high expression of ubiquitin in the retina. Analysis of lipofuscin fluorophores in the RPE showed a significant elevation of A2E and compounds of the all-trans-retinal dimer series in retinas from four and ten month old E_mut(+/-) mice compared to wild-type littermates. These observations suggest that abnormal accumulation of ELOVL4 protein and lipofuscin may lead to photoreceptor degeneration in E_mut(+/-) mice. PMID- 19682987 TI - Interleukin-1beta-induced brain injury in the neonatal rat can be ameliorated by alpha-phenyl-n-tert-butyl-nitrone. AB - To examine the possible role of inflammatory cytokines in mediating perinatal brain injury, we investigated effects of intracerebral injection of interleukin 1beta (IL-1beta) on brain injury in the neonatal rat and the mechanisms involved. Intracerebral administration of IL-1beta (1 microg/kg) resulted in acute brain injury, as indicated by enlargement of ventricles bilaterally, apoptotic death of oligodendrocytes (OLs) and loss of OL immunoreactivity in the neonatal rat brain. IL-1beta also induced axonal and neuronal injury in the cerebral cortex as indicated by elevated expression of beta-amyloid precursor protein, short beaded axons and dendrites, and loss of tyrosine hydroxylase-positive neurons in the substantia nigra and the ventral tegmental areas. Administration of alpha-phenyl n-tert-butyl-nitrone (PBN, 100 mg/kg i.p.) immediately after the IL-1beta injection protected the brain from IL-1beta-induced injury. Protection of PBN was linked with the attenuated oxidative stress induced by IL-1beta, as indicated by decreased elevation of 8-isoprostane content and by the reduced number of 4 hydroxynonenal or malondialdehyde or nitrotyrosine-positive cells following IL 1beta exposure. PBN also attenuated IL-1beta-stimulated inflammatory responses as indicated by the reduced activation of microglia. The finding that IL-1beta induced perinatal brain injury was very similar to that induced by lipopolysaccharide (LPS), as we previously reported and that PBN was capable to attenuate the injury induced by either LPS or IL-1beta suggests that IL-1beta may play a critical role in mediating brain injury associated with perinatal infection/inflammation. PMID- 19682988 TI - Natural lipophilic inhibitors of mitochondrial complex I are candidate toxins for sporadic neurodegenerative tau pathologies. AB - Annonacin, a natural lipophilic inhibitor of mitochondrial complex I has been implicated in the etiology of a sporadic neurodegenerative tauopathy in Guadeloupe. We therefore studied further compounds representing the broad biochemical spectrum of complex I inhibitors to which humans are potentially exposed. We determined their lipophilicity, their effect on complex I activity in submitochondrial particles, and their effect on cellular ATP levels, neuronal cell death and somatodendritic redistribution of phosphorylated tau protein (AD2 antibody against pS396/pS404-tau) in primary cultures of fetal rat striatum. The 24 compounds tested were lipophilic (logP range 0.9-8.5; exception: MPP(+) logP= 1.35) and potent complex I inhibitors (IC(50) range 0.9 nM-2.6 mM). They all decreased ATP levels (EC(50) range 1.9 nM-54.2 microM), induced neuronal cell death (EC(50) range 1.1 nM-54.5 microM) and caused the redistribution of AD2(+) tau from axons to the cell body (EC(5) range 0.6 nM-33.3 microM). The potency of the compounds to inhibit complex I correlated with their potency to induce tau redistribution (r=0.80, p<0.001). In conclusion, we propose that the widely distributed lipophilic complex I inhibitors studied here might be implicated in the induction of tauopathies with global prevalence. PMID- 19682989 TI - Mesenchymal stem cell transplantation in amyotrophic lateral sclerosis: A Phase I clinical trial. AB - Amyotrophic Lateral Sclerosis (ALS) is a devastating incurable disease. Stem-cell based therapies represent a new possible strategy for ALS clinical research. The objectives of this Phase 1 clinical study were to assess the feasibility and toxicity of mesenchymal stem cell transplantation and to test the impact of a cell therapy in ALS patients. The trial was approved and monitored by the National Institute of Health and by the Ethics Committees of all participating Institutions. Autologous MSCs were isolated from bone marrow, expanded in vitro and analyzed according to GMP conditions. Expanded MSCs were suspended in the autologous cerebrospinal fluid (CSF) and directly transplanted into the spinal cord at a high thoracic level with a surgical procedure. Ten ALS patients were enrolled and regularly monitored before and after transplantation by clinical, psychological, neuroradiological and neurophysiological assessments. There was no immediate or delayed transplant-related toxicity. Clinical, laboratory, and radiographic evaluations of the patients showed no serious transplant-related adverse events. Magnetic resonance images (MRI) showed no structural changes (including tumor formation) in either the brain or the spinal cord. However the lack of post mortem material prevents any definitive conclusion about the vitality of the MSCs after transplantation. In conclusion, this study confirms that MSC transplantation into the spinal cord of ALS patients is safe and that MSCs might have a clinical use for future ALS cell based clinical trials. PMID- 19682990 TI - Engraftment of embryonic stem cell-derived myogenic progenitors in a dominant model of muscular dystrophy. AB - Muscular dystrophies (MDs) consist of a genetically heterogeneous group of disorders, recessive or dominant, characterized by progressive skeletal muscle weakening. To date, no effective treatment is available. Experimental strategies pursuing muscle regeneration through the transplantation of stem cell preparations have brought hope to patients affected by this disorder. Efficacy has been demonstrated in recessive MD models through contribution of wild-type nuclei to the muscle fiber heterokaryon; however, to date, there has been no study investigating the efficacy of a cell therapy in a dominant model of MD. We have recently demonstrated that Pax3-induced embryonic stem (ES) cell-derived myogenic progenitors are able to engraft and improve muscle function in mdx mice, a recessive mouse model for Duchenne MD. To assess whether this therapeutic effect can be extended to a dominant type of muscle disorder, here we transplanted these cells into FRG1 transgenic mice, a dominant model that has been associated with facioscapulohumeral muscular dystrophy. Our results show that Pax3-induced ES-derived myogenic progenitors are capable of significant engraftment after intramuscular or systemic transplantation into Frg1 mice. Analyses of contractile parameters revealed functional improvement in treated muscles of male mice, but not females, which are less severely affected. This study is the first to use Frg1 transgenic mice to assess muscle regeneration as well as to support the use of a cell-based therapy for autosomal dominant types of MD. PMID- 19682991 TI - Prenatal blockade of Ang II receptors affects neonatal rat hindbrain structure and receptor localization. AB - The development of knock-out mice for Angiotensin II (Ang II) AT(2) receptors, which exhibited altered exploratory behavior, prompted us to investigate the cerebellum and brainstem. We evaluated the effect of stimulation/inhibition of Ang II receptors on hindbrain development, in offspring (postnatal days P0, P8) of pregnant rats treated during late pregnancy (Ang II, Losartan or PD123319, 1 mg/kg/day). Receptor localization by autoradiography showed in P0 and P8 hindbrains, that most structures expressed AT(2) subtype: cerebellar cortex, cerebellar nuclei, genu facial nucleus, inferior colicullus, inferior olive. In the cerebellar cortex, [(125)I]Ang II AT(2) binding was predominant, while low AT(1) binding was observed in adjacent layers of the cerebellar cortex. Blockade of AT(2) receptors with PD123319 increased binding in cerebellar nuclei (p<0.05) and brainstem nuclei at P0, P8, in correlation with increased AT(2) receptor expression by RT-PCR. The enlarged external granular layer (EGL) in PD123319 treated P0 pups contrast with the significant decrease in Ang II binding (p<0.001) in the cerebellar cortex. Blockade of AT(2) receptors during late pregnancy seems to arrest cerebellar cortex development in P0 animals. On the contrary, increased AT(2) binding was observed in cerebellar cortex and DTg nucleus in PD123319-treated P8 animals (p<0.001). Ang II treatment leads to increased binding in the brainstem. In spite of the low doses of Ang II antagonists used, treatments were performed during a time-frame critical for hindbrain development, leading to remarkable effects. The present study makes a contribution to understand the role of Ang II receptors during hindbrain development. PMID- 19682992 TI - Jid1 is a J-protein functioning in the mitochondrial matrix, unable to directly participate in endoplasmic reticulum associated protein degradation. AB - J-proteins are a class of molecular chaperones that serve to stimulate the activity of Hsp70s and are often located to recruit Hsp70 to a particular cellular function. Protein degradation associated with the endoplasmic reticulum (ERAD) is one such cellular process that requires Hsp70 on both faces of the endoplasmic reticulum. At least five J-proteins, including Jid1 (DnaJ protein Involved in ER-associated Degradation), have been implicated in controlling ERAD. Here we show that Jid1 is confined within the mitochondrial matrix - Jid1 has the same topology as the J-proteins Pam18 and Mdj2, which stimulate mitochondrial Hsp70 to drive protein import into the mitochondrial matrix. The location of Jid1 within mitochondria makes it unavailable to participate directly in the regulation of ERAD. PMID- 19682993 TI - A single exposure to an acute stressor has lasting consequences for the hypothalamo-pituitary-adrenal response to stress in free-living birds. AB - In vertebrates, activation of the hypothalamo-pituitary-adrenal (HPA) axis in response to unpredictable events results in elevated glucocorticoid secretion. Repeated exposure to stressors alters subsequent glucocorticoid secretion, either by inducing chronic stress or as a result of habituation. However, most studies of repeated stress focus on the impacts of multiple and frequent exposures to acute stressors, and few have been carried out in free-living animals. We investigated whether a single exposure to a novel stressor was sufficient to produce long-lasting alterations in HPA function in free-living eastern bluebirds (Sialia sialis). We subjected adult females to a capture/restraint protocol in which we collected serial blood samples over an hour of restraint to be analyzed for corticosterone. We administered this protocol to three groups of females during the nestling phase of their first and/or second brood of the season: Repeaters (sampled during brood 1 and brood 2), Naive-Brood 1 (sampled only during brood 1), and Naive-Brood 2 (sampled only during brood 2). Repeaters had attenuated corticosterone responses to the second restraint bout compared to the first, and in brood 2, Repeaters had lower responses than Naive-Brood 2 females. However, Naive-Brood 1 and Naive-Brood 2 birds did not differ in their responses to restraint. Thus, as little as one prior experience with an acute stressor was sufficient to alter subsequent HPA responsiveness, and this effect was not due to a natural change in HPA responsiveness as the breeding season progressed. These data may have important implications for understanding how acute stressors can alter a free-living animal's ability to cope in the face of subsequent stressors, and for longitudinal field studies in which individuals are repeatedly sampled for glucocorticoid responsiveness. PMID- 19682994 TI - Development of highly sensitive Bicistronic vector based non-radioactive antigen specific cytotoxicity assay. AB - In the absence of a better alternate, (51)Cr release assay, with its several disadvantages is still the most common method for detection of MHC class I restricted T-cell mediated cytotoxicity. We describe a system in which the T-cell mediated cytotoxicity can be assessed using host-derived cells transfected with a bicistronic vector expressing the specific antigen and a quantifiable reporter as target cells. This overcomes the problems associated with use of radioactivity, pre-loading of target cells with reporter/antigen and the MHC restriction. We used HBV core antigen to prove the concept, as it is an established CTL target. Bicistronic vectors containing HBV core and reporter (EGFP/Fluc) gene were generated and further checked for antigen/reporter expression in human HepG2, mouse fibroblast BALB/c.3T3 and mouse lymphoma A20 cell lines. The effector cells to study the cytolytic activity were generated in vivo using BALB/c mice immunized with antigen expressing DNA clone or protein. The target cells (BALB/c.3T3 and A20) transiently transfected with bicistronic constructs were incubated with effector cells (splenocytes) from immunized mice at a different effector to target ratio. Following incubation the CTL activity was calculated by measuring the reporter luciferase in the remaining viable target cells that inversely correlates with the cytolysis of susceptible cells. The percent specific lysis measured in our assay was compared with conventional (51)Cr release assay to validate this approach. This novel bicistronic vector based cytotoxicity assay demonstrated an easy to perform, antigen-specific and non radioactive method of determining T-cell mediated cytotoxicity. PMID- 19682995 TI - DNA sensor development based on multi-wall carbon nanotubes for label-free influenza virus (type A) detection. AB - This paper describes the DNA immobilization using carbon multi-walled nanotubes (MWCNTs) for direct and label-free detection of influenza virus (type A). The DNA probe was attached on the sensor surface by means of covalent bonding between the amine and phosphate groups of the DNA sequence. The interaction between the DNA probe and the MWCNTs were characterized by Fourier Transform Infrared (FTIR) spectrometry, Raman spectra. The hybridization of the DNA probe and the target DNA were detected by changes in the conductance on the surface of sensors leading to the change in the output signal of the system. The results show that the DNA sensor can detect as low as 0.5 nM of the target DNA samples; the response time of DNA sensor is approximately 4 min. PMID- 19682996 TI - Critical oxygen partial pressures and maximal tracheal conductances for Drosophila melanogaster reared for multiple generations in hypoxia or hyperoxia. AB - In Drosophila melanogaster and other insects, increases in atmospheric oxygen partial pressure (aPO(2)) tend to increase adult body size and decrease tracheal diameters and tracheolar proliferation. If changes in tracheal morphology allow for functional compensation for aPO(2), we would predict that higher aPO(2) would be associated with higher critical PO(2) values (CritPO(2)) and lower maximal tracheal conductances (G(max)). We measured CritPO(2) and G(max) for adult and larval vinegar flies reared for 7-9 generations in 10, 21 or 40 kPa O(2). The CritPO(2), CO(2) emission rates and G(max) values were generally independent of the rearing PO(2) these flies had experienced, suggesting that minimal functional changes in tracheal capacities occurred in response to rearing PO(2). Larvae were able to continue activity during 20 min of anoxia. The lack of multigenerational rearing PO(2) effects on tracheal function suggests that the functional compensation at the whole-body level due to tracheal morphological changes in response to aPO(2) may be minimal; alternatively the benefits of such compensation may occur in specific tissues or during processes not assessed by these methods. In larvae, the CritPO(2) and the capacity for movement in anoxia suggest adaptations for life in hypoxic organic matter. PMID- 19682997 TI - Effect of macromolecular crowding on protein folding dynamics at the secondary structure level. AB - Macromolecular crowding is one of the key characteristics of the cellular environment and is therefore intimately coupled to the process of protein folding in vivo. While previous studies have provided invaluable insight into the effect of crowding on the stability and folding rate of protein tertiary structures, very little is known about how crowding affects protein folding dynamics at the secondary structure level. In this study, we examined the thermal stability and folding-unfolding kinetics of three small folding motifs (i.e., a 34-residue alpha-helix, a 34-residue cross-linked helix-turn-helix, and a 16-residue beta hairpin) in the presence of two commonly used crowding agents, Dextran 70 (200 g/L) and Ficoll 70 (200 g/L). We found that these polymers do not induce any appreciable changes in the folding kinetics of the two helical peptides, which is somewhat surprising as the helix-coil transition kinetics have been shown to depend on viscosity. Also to our surprise and in contrast to what has been observed for larger proteins, we found that crowding leads to an appreciable decrease in the folding rate of the shortest beta-hairpin peptide, indicating that besides the excluded volume effect, other factors also need to be considered when evaluating the net effect of crowding on protein folding kinetics. A model considering both the static and the dynamic effects arising from the presence of the crowding agent is proposed to rationalize these results. PMID- 19682998 TI - Conformational flexibility in immunoglobulin E-Fc 3-4 revealed in multiple crystal forms. AB - The structure of immunoglobulin E (IgE)-Fc(3-4) has been solved in three new crystal forms, providing 13 snapshots of the Fc conformation and revealing a diverse range of open-closed motions among subunit chains and dimers. A more detailed analysis of the open-to-closed motion of IgE-Fc(3-4) was possible with so many structures, and the new structures allow a more thorough examination of the flexibility of IgE-Fc and its implications for receptor binding. The existence of a hydrophobic pocket at the elbow region of the Fc appears to be conformation dependent and suggests a means of regulating the IgE-Fc conformation (and potentially receptor binding) with small molecules. PMID- 19682999 TI - On the divalent metal ion dependence of DNA cleavage by restriction endonucleases of the EcoRI family. AB - Restriction endonucleases of the PD...D/EXK family need Mg(2+) for DNA cleavage. Whereas Mg(2+) (or Mn(2+)) promotes catalysis, Ca(2+) (without Mg(2+)) only supports DNA binding. The role of Mg(2+) in DNA cleavage by restriction endonucleases has elicited many hypotheses, differing mainly in the number of Mg(2+) involved in catalysis. To address this problem, we measured the Mg(2+) and Mn(2+) concentration dependence of DNA cleavage by BamHI, BglII, Cfr10I, EcoRI, EcoRII (catalytic domain), MboI, NgoMIV, PspGI, and SsoII, which were reported in co-crystal structure analyses to bind one (BglII and EcoRI) or two (BamHI and NgoMIV) Me(2+) per active site. DNA cleavage experiments were carried out at various Mg(2+) and Mn(2+) concentrations at constant ionic strength. All enzymes show a qualitatively similar Mg(2+) and Mn(2+) concentration dependence. In general, the Mg(2+) concentration optimum (between approximately 1 and 10 mM) is higher than the Mn(2+) concentration optimum (between approximately 0.1 and 1 mM). At still higher Mg(2+) or Mn(2+) concentrations, the activities of all enzymes tested are reduced but can be reactivated by Ca(2+). Based on these results, we propose that one Mg(2+) or Mn(2+) is critical for restriction enzyme activation, and binding of a second Me(2+) plays a role in modulating the activity. Steady-state kinetics carried out with EcoRI and BamHI suggest that binding of a second Mg(2+) or Mn(2+) mainly leads to an increase in K(m), such that the inhibitory effect of excess Mg(2+) or Mn(2+) can be overcome by increasing the substrate concentration. Our conclusions are supported by molecular dynamics simulations and are consistent with the structural observations of both one and two Me(2+) binding to these enzymes. PMID- 19683000 TI - Mapping transient partial unfolding by protein engineering and native-state proteolysis. AB - Transient partial unfolding of proteins under native conditions may have significant consequences in the biochemical and biophysical properties of proteins. Native-state proteolysis offers a facile way to investigate the thermodynamic and kinetic accessibilities of partially unfolded forms (cleavable forms) under native conditions. However, determination of the structure of the cleavable form, which is populated only transiently, remains challenging. Although in some cases partially cleaved products from proteolysis provide information on the structure of this elusive form, proteolysis of many proteins does not accumulate detectable intermediates. Here, we describe a systematic approach to determining structures of cleavable forms by protein engineering and native-state proteolysis. By devising phi(c) analysis, which is analogous to conventional phi analysis, we have determined the structure of the cleavable form of Escherichia coli maltose-binding protein (MBP), which does not accumulate any partially cleaved products. We mutated 10 buried residues in MBP to alanine and determined phi(c) values from the effects of the mutations on global stability and proteolytic susceptibility. The result of this analysis suggests that two C terminal helices in MBP are unfolded in their cleavable form. The effect of ligand binding on proteolytic susceptibility and C-terminal deletion mutations also confirms the proposed structure. Our approach and methodology are generally applicable not only in elucidating the mechanism of proteolysis but also in investigating other important processes involving partial unfolding under native conditions such as protein misfolding and aggregation. PMID- 19683001 TI - A mechanism for histone chaperoning activity of nucleoplasmin: thermodynamic and structural models. AB - Nucleoplasmin (NP), a histone chaperone, acts as a reservoir for histones H2A-H2B in Xenopus laevis eggs and can displace sperm nuclear basic proteins and linker histones from the chromatin fiber of sperm and quiescent somatic nuclei. NP has been proposed to mediate the dynamic exchange of histones during the expression of certain genes and assists the assembly of nucleosomes by modulating the interaction between histones and DNA. Here, solution structural models of full length NP and NP complexes with the functionally distinct nucleosomal core and linker histones are presented for the first time, providing a picture of the physical interactions between the nucleosomal and linker histones with NP core and tail domains. Small-angle X-ray scattering and isothermal titration calorimetry reveal that NP pentamer can accommodate five histones, either H2A-H2B dimers or H5, and that NP core and tail domains are intimately involved in the association with histones. The analysis of the binding events, employing a site specific cooperative model, reveals a negative cooperativity-based regulatory mechanism for the linker histone/nucleosomal histone exchange. The two histone types bind with drastically different intrinsic affinity, and the strongest affinity is observed for the NP variant that mimicks the hyperphosphorylated active protein. The different "affinity windows" for H5 and H2A-H2B might allow NP to fulfill its histone chaperone role, simultaneously acting as a reservoir for the core histones and a chromatin decondensing factor. Our data are compatible with the previously proposed model where NP facilitates nucleosome assembly by removing the linker histones and depositing H2A-H2B dimers onto DNA. PMID- 19683002 TI - Crystal structures of glycosyltransferase UGT78G1 reveal the molecular basis for glycosylation and deglycosylation of (iso)flavonoids. AB - The glycosyltransferase UGT78G1 from Medicago truncatula catalyzes the glycosylation of various (iso)flavonoids such as the flavonols kaempferol and myricetin, the isoflavone formononetin, and the anthocyanidins pelargonidin and cyanidin. It also catalyzes a reverse reaction to remove the sugar moiety from glycosides. The structures of UGT78G1 bound with uridine diphosphate or with both uridine diphosphate and myricetin were determined at 2.1 A resolution, revealing detailed interactions between the enzyme and substrates/products and suggesting a distinct binding mode for the acceptor/product. Comparative structural analysis and mutagenesis identify glutamate 192 as a key amino acid for the reverse reaction. This information provides a basis for enzyme engineering to manipulate substrate specificity and to design effective biocatalysts with glycosylation and/or deglycosylation activity. PMID- 19683003 TI - DARPins as bispecific receptor antagonists analyzed for immunoglobulin E receptor blockage. AB - The concept of multispecific antibodies is of high therapeutic interest but has failed to produce pharmaceutical products due to the poor biophysical properties of such molecules. Here, we propose an alternative and simple way to generate bispecific binding molecules using designed ankyrin repeat proteins (DARPins). For this purpose, monovalent DARPins with different epitope specificities were selected against the alpha chain of the high-affinity receptor for human immunoglobulin E (IgE) (FcepsilonRIalpha). Two of the isolated binders interfering with IgE binding to the receptor were joined to each other or to themselves via a flexible protein linker. The resulting bivalent and bispecific DARPins were tested for their ability to prevent allergen-induced cell degranulation using rat basophilic leukemia cells stably transfected with human FcepsilonRIalpha. The bispecific DARPin construct was the most potent one, efficiently blocking the IgE-FcepsilonRI interaction and preventing the release of proinflammatory mediators. Noteworthy, the multivalent and multispecific DARPin construct did not show any alteration of the beneficial biophysical properties of the monovalent parental DARPins. Hence, bispecific DARPins may be used to generate receptor antagonists simultaneously targeting different epitopes on the same molecule. Moreover, they easily overcome the limiting immunoglobulin binding paradigm (one binding molecule=one epitope) and thereby represent an alternative to monoclonal antibodies in cases where the immunoglobulin scaffold is unsuitable. PMID- 19683004 TI - Structural basis of conserved cysteine in the fibroblast growth factor family: evidence for a vestigial half-cystine. AB - The 22 members of the mouse/human fibroblast growth factor (FGF) family of proteins contain a conserved cysteine residue at position 83 (numbering scheme of the 140-residue form of FGF-1). Sequence and structure information suggests that this position is a free cysteine in 16 members and participates as a half-cystine in at least 3 (and perhaps as many as 6) other members. While a structural role as a half-cystine provides a stability basis for possible selective pressure, it is less clear why this residue is conserved as a free cysteine (although free buried thiols can limit protein functional half-life). To probe the structural role of the free cysteine at position 83 in FGF-1, we constructed Ala, Ser, Thr, Val, and Ile mutations and determined their effects on structure and stability. These results show that position 83 in FGF-1 is thermodynamically optimized to accept a free cysteine. A second cysteine mutation was introduced into wild-type FGF-1 at adjacent position Ala66, which is known to participate as a half-cystine with position 83 in FGF-8, FGF-19, and FGF-23. Results show that, unlike position 83, a free cysteine at position 66 destabilizes FGF-1; however, upon oxidation, a near-optimal disulfide bond is formed between Cys66 and Cys83, resulting in approximately 14 kJ/mol of increased thermostability. Thus, while the conserved free cysteine at position 83 in the majority of the FGF proteins may have a principal role in limiting functional half-life, evidence suggests that it is a vestigial half-cystine. PMID- 19683005 TI - Identification of a mechanical rheostat in the hydrophobic core of protein L. AB - The ability of proteins and their complexes to withstand or respond to mechanical stimuli is vital for cells to maintain their structural organisation, to relay external signals and to facilitate unfolding and remodelling. Force spectroscopy using the atomic force microscope allows the behaviour of single protein molecules under an applied extension to be investigated and their mechanical strength to be quantified. protein L, a simple model protein, displays moderate mechanical strength and is thought to unfold by the shearing of two mechanical sub-domains. Here, we investigate the importance of side-chain packing for the mechanical strength of protein L by measuring the mechanical strength of a series of protein L variants containing single conservative hydrophobic volume deletion mutants. Of the five thermodynamically destabilized variants characterised, only one residue (I60 V) close to the interface between two mechanical sub-domains was found to differ in mechanical properties to wild type (Delta F(I60 V-WT)=-36 pN at 447 nm s(-1), Delta x(uI60V-WT)=0.2 nm). Phi-value analysis of the unfolding data revealed a highly native transition state. To test whether the number of hydrophobic contacts across the mechanical interface does affect the mechanical strength of protein L, we measured the mechanical properties of two further variants. protein L L10F, which increases core packing but does not enhance interfacial contacts, increased mechanical strength by 13+/-11 pN at 447 nm s( 1). By contrast, protein L I60F, which increases both core and cross-interface contacts, increased mechanical strength by 72+/-13 pN at 447 nm s(-1). These data suggest a method by which nature can evolve a varied mechanical response from a limited number of topologies and demonstrate a generic but facile method by which the mechanical strength of proteins can be rationally modified. PMID- 19683007 TI - Nonnative electrostatic interactions can modulate protein folding: molecular dynamics with a grain of salt. AB - In recent years, a growing number of protein folding studies have focused on the unfolded state, which is now recognized as playing a major role in the folding process. Some of these studies show that interactions occurring in the unfolded state can significantly affect the stability and kinetics of the protein folding reaction. In this study, we modeled the effect of electrostatic interactions, both native and nonnative, on the folding of three protein systems that underwent selective charge neutralization or reversal or complete charge suppression. In the case of the N-terminal L9 protein domain, our results directly attribute the increase in thermodynamic stability to destabilization of the unfolded ensemble, reaffirming the experimental observations. These results provide a deeper structural insight into the ensemble of the unfolded state and predict a new mutation site for increased protein stability. In the second case, charge reversal mutations of RNase Sa affected protein stability, with the destabilizing mutations being less destabilizing at higher salt concentrations, indicating the formation of charge-charge interactions in the unfolded state. In the N-terminal L9 and RNase Sa systems, changes in electrostatic interactions in the unfolded state that cause an increase in free energy had an overall compaction effect that suggests a decrease in entropy. In the third case, in which we compared the beta lactalbumin and hen egg-white lysozyme protein homologues, we successfully eliminated differences between the folding kinetics of the two systems by suppressing electrostatic interactions, supporting previously reported findings. Our coarse-grained molecular dynamics study not only reproduces experimentally reported findings but also provides a detailed molecular understanding of the elusive unfolded-state ensemble and how charge-charge interactions can modulate the biophysical characteristics of folding. PMID- 19683006 TI - Adaptive evolution of p53 thermodynamic stability. AB - The thermodynamic stability of a protein plays an important role during evolution and adaptation in order to maintain a folded and active conformation. p53 is a tumour suppressor involved in the regulation of numerous genes. Human p53 has an unusually low thermodynamic stability and is frequently inactivated by oncogenic missense mutations. Here, we examined the thermodynamic and kinetic stability of p53 DNA binding domains from selected invertebrate and vertebrate species by differential scanning calorimetry and equilibrium urea denaturation. There is a correlation in the apparent melting temperature of p53 with the body temperature of homeotherm vertebrates. We found that p53 from these organisms has a half-life for spontaneous unfolding at organismal body temperature of 10-20 min. We also found that p53 from invertebrates has higher stability, bearing more resemblance towards p63 and p73 from humans. Using structure-guided mutagenesis on the human p53 scaffold, we demonstrated that the amino acid changes on the protein surface and in the protein interior lead to the elevated stability of p53 orthologs. We propose a model in which the p53 DNA binding domain has been shaped by the complex interplay of different selective pressures and underwent adaptive evolution leading to pronounced effects on its stability. p53 from vertebrates has evolved to have a low thermodynamic stability and similarly short spontaneous half-life at organismal body temperature, which is related to function. PMID- 19683010 TI - Sustainability of dioecious and hermaphrodite populations on a lattice. AB - Sexual reproduction may be divided into two main categories: hermaphroditism and dioecy (Botany)/gonochorism (Zoology). Simultaneous hermaphrodites can function in both male and female roles whereas a dioecious/gonochorist population consists of distinct male and female individuals. Mean-field calculations, which ignore spatial aspects, suggest that self-incompatible hermaphrodites should have a twofold advantage over dioecious population when reproduction is limited by mating encounters. By use of stochastic spatial simulations we demonstrate that hermaphroditism has an even greater advantage when local interactions are considered. This result provides further support for the observation that hermaphroditism is associated with sedentary species, such as plants and animals with poor mate search efficiency. We also investigate the finite size effects associated with the well-known quadratic contact process. PMID- 19683008 TI - Altered contractility of skeletal muscle in mice deficient in titin's M-band region. AB - We investigated the contractile phenotype of skeletal muscle deficient in exons MEx1 and MEx2 (KO) of the titin M-band by using the cre-lox recombination system and a multidisciplinary physiological approach to study skeletal muscle contractile performance. At a maximal tetanic stimulation frequency, intact KO extensor digitorum longus muscle was able to produce wild-type levels of force. However, at submaximal stimulation frequency, force was reduced in KO mice, giving rise to a rightward shift of the force-frequency curve. This rightward shift of the force-frequency curve could not be explained by altered sarcoplasmic reticulum Ca(2+) handling, as indicated by analysis of Ca(2+) transients in intact myofibers and expression of Ca(2)(+)-handling proteins, but can be explained by the reduced myofilament Ca(2+) sensitivity of force generation that we found. Western blotting experiments suggested that the excision of titin exons MEx1 and MEx2 did not result in major changes in expression of titin M-band binding proteins or phosphorylation level of the thin-filament regulatory proteins, but rather in a shift toward expression of slow isoforms of the thick filament-associated protein, myosin binding protein-C. Extraction of myosin binding protein-C from skinned muscle normalized myofilament Ca(2+) sensitivity of the KO extensor digitorum longus muscle. Thus, our data suggest that the M band region of titin affects the expression of genes involved in the regulation of skeletal muscle contraction. PMID- 19683011 TI - Environmentally induced mechanical feedback in locomotion: frog performance as a model. AB - At first glance, the strategy for generating propulsive impulses for both jumping and swimming in frogs is quite similar. Both modes rely on powerful extension of the hind limbs. However, in Rana esculenta (the semi-aquatic green frog), propulsive impulses for jumping were found to be much larger than those generated during swimming [Nauwelaerts and Aerts, 2003. Propulsive impulses as a covarying performance measure in the comparison of the kinematics of swimming and jumping in frogs. J. Exp. Biol. 206, 4341-4351]. The hypothesis that differences in propulsive impulse between swimming and jumping are largely caused by specific environmental constraints rather than being due to changes in motor control is tested in the present study. To assess this question, the actuator of a simple mathematical model, mimicking a frog with symmetrically kicking hind limbs, is first tuned to perform frog-like jumps. Next, the same actuator activation is applied to drive the model in an 'aquatic environment'. Despite the entirely identical activation, the resulting in silico propulsive swimming impulse was less than half that produced during jumping, just as observed in vivo. Although duration of limb extension is similar for both locomotor modes (both in vivo and in silico), this conspicuous difference in model behaviour is entirely explained by the actuator working at different positions along its force-velocity curve. These findings suggest that the same environmentally induced effects are also involved in real swimming and jumping as well, thus explaining the apparent difference in performance level. PMID- 19683009 TI - Structural redesign of lipase B from Candida antarctica by circular permutation and incremental truncation. AB - Circular permutation of Candida antarctica lipase B yields several enzyme variants with substantially increased catalytic activity. To better understand the structural and functional consequences of protein termini reorganization, we have applied protein engineering and x-ray crystallography to cp283, one of the most active hydrolase variants. Our initial investigation has focused on the role of an extended surface loop, created by linking the native N- and C-termini, on protein integrity. Incremental truncation of the loop partially compensates for observed losses in secondary structure and the permutants' temperature of unfolding. Unexpectedly, the improvements are accompanied by quaternary-structure changes from monomer to dimer. The crystal structures of one truncated variant (cp283 Delta 7) in the apo-form determined at 1.49 A resolution and with a bound phosphonate inhibitor at 1.69 A resolution confirmed the formation of a homodimer by swapping of the enzyme's 35-residue N-terminal region. Separately, the new protein termini at amino acid positions 282/283 convert the narrow access tunnel to the catalytic triad into a broad crevice for accelerated substrate entry and product exit while preserving the native active-site topology for optimal catalytic turnover. PMID- 19683012 TI - Assessment of weight bearing changes and pharmacological antinociception in mice with LPS-induced monoarthritis using the Catwalk gait analysis system. AB - AIMS: We evaluated the possibility of using the video-based Catwalk gait analysis method to measure weight bearing changes and for testing pharmacological antinociception in freely moving mice with lipopolysaccharide (LPS)-induced monoarthritis. MAIN METHODS: LPS or its solvent (PBS) was injected intra articularly into the right hind (RH) limb ankle joint through the Achilles tendon of C57BL/6 mice. The Catwalk system was used to assess behavioral changes in freely moving mice. The effects of indomethacin on changes in LPS-inoculated mice were examined. KEY FINDINGS: Mice inoculated with LPS into the RH limb showed reduced paw pressure (measured as light intensity) and print area on the RH limb, whereas they exerted more pressure with the left hind (LH) and front limbs, showing a transfer of weight bearing from RH to LH and front limbs, which was significant at 2 days post-LPS inoculation. There were no differences between the front limbs. No changes were observed in the PBS injected controls. There were no changes in interlimb coordination (regularity index) in both PBS- and LPS injected mice. Treatment with indomethacin (10 and 100mg/kg) restored the weight bearing (measured as the ratio of the pressure exerted by the paws) and the print area ratios of LPS-inoculated mice similar to that observed in control mice. SIGNIFICANCE: This study shows that the Catwalk gait analysis system can be used to objectively quantify LPS-induced monoarthritis weight bearing changes in all four limbs and evaluate pharmacological antinociception in freely moving mice. PMID- 19683014 TI - Decentralized resource allocation to control an epidemic: a game theoretic approach. AB - This paper examines how two countries would allocate resources at the onset of an epidemic when they seek to protect their own populations by minimizing the total number of infectives over the entire time horizon. We model this situation as a game between selfish countries, where players strategically allocate their resources in order to minimize the total number of infected individuals in their respective populations during the epidemic. We study this problem when the initial number of infectives is very small, which greatly simplifies the analysis. We show in this framework that selfish countries always allocate their resources so as to bring the effective reproduction ratio below one and avoid a major outbreak. When a major outbreak is avoidable, we further identify the necessary and sufficient conditions under which the individual allocation decisions of selfish countries match the decision that a central planner would make in order to minimize the total number of infectives in the whole population (without distinguishing between countries). PMID- 19683013 TI - Dose dependent development of diabetes mellitus and non-alcoholic steatohepatitis in monosodium glutamate-induced obese mice. AB - AIMS: We have recently reported that monosodium glutamate (MSG) induces severe obesity with diabetes mellitus and/or non-alcoholic fatty liver disease (NAFLD)/non-alcoholic steatohepatitis (NASH) in Crj:CD-1(ICR) neonatal mice. In this study, we investigated the effects of varying the dose of MSG on the resulting obesity and diabetes mellitus. MAIN METHODS: Crj:CD-1(ICR) neonatal mice were administered MSG in one of several courses: once-daily subcutaneous injections of 2mg/g for 5 consecutive days (2 mg/g x 5 group), a single subcutaneous injection of 4 mg/g (4 mg/g x 1 group) and once-daily subcutaneous injections of 4 mg/g for 5 consecutive days (4 mg/g x 5 group). KEY FINDINGS: In all the MSG treatment groups, severe obesity developed by 29 weeks of age. The onset of diabetes mellitus and liver lesions (resembling those of human NAFLD/NASH) were observed before 54 weeks of age. The obesity, diabetes mellitus and liver lesions were most severe in the 4 mg/g x 1 group. In the 4 mg/g x 5 group, increases in body weight and body length were inhibited by MSG's severe toxicity. SIGNIFICANCE: A single 4 mg/g dose of MSG is the most suitable as the obese model and induces not only severe obesity and diabetes mellitus, but also liver changes resembling human NAFLD/NASH. A small amount of MSG in the newborn develops obesity and the other complications without hyperphagia after a long term. PMID- 19683015 TI - Dynamic phenomena arising from an extended Core Group model. AB - In order to obtain a reasonably accurate model for the spread of a particular infectious disease through a population, it may be necessary for this model to possess some degree of structural complexity. Many such models have, in recent years, been found to exhibit a phenomenon known as backward bifurcation, which generally implies the existence of two subcritical endemic equilibria. It is often possible to refine these models yet further, and we investigate here the influence such a refinement may have on the dynamic behaviour of a system in the region of the parameter space near R(0)=1. We consider a natural extension to a so-called Core Group model for the spread of a sexually transmitted disease, arguing that this may in fact give rise to a more realistic model. From the deterministic viewpoint we study the possible shapes of the resulting bifurcation diagrams and the associated stability patterns. Stochastic versions of both the original and the extended models are also developed so that the probability of extinction and time to extinction may be examined, allowing us to gain further insights into the complex system dynamics near R(0)=1. A number of interesting phenomena are observed, for which heuristic explanations are provided. PMID- 19683016 TI - Microsaccades: small steps on a long way. AB - Contrary to common wisdom, fixations are a dynamically rich behavior, composed of continual, miniature eye movements, of which microsaccades are the most salient component. Over the last few years, interest in these small movements has risen dramatically, driven by both neurophysiological and psychophysical results and by advances in techniques, analysis, and modeling of eye movements. The field has a long history but a significant portion of the earlier work has gone missing in the current literature, in part, as a result of the collapse of the field in the 1980s that followed a series of discouraging results. The present review compiles 60 years of work demonstrating the unique contribution of microsaccades to visual and oculomotor function. Specifically, the review covers the contribution of microsaccades to (1) the control of fixation position, (2) the reduction of perceptual fading and the continuity of perception, (3) the generation of synchronized visual transients, (4) visual acuity, (5) scanning of small spatial regions, (6) shifts of spatial attention, (7) resolving perceptual ambiguities in the face of multistable perception, as well as several other functions. The accumulated evidence demonstrates that microsaccades serve both perceptual and oculomotor goals and although in some cases their contribution is neither necessary nor unique, microsaccades are a malleable tool conveniently employed by the visual system. PMID- 19683017 TI - Fine-tuning the brain: MicroRNAs. AB - The brain is of bewildering complexity and numerous genes and signaling molecules have been described that affect the architecture and functioning of specific neuronal circuits. Recent evidence from genome analysis revealed the existence of a large group of novel RNA molecules with unexpected properties. One such group is called microRNAs, which are small 21-23 nucleotides RNA molecules that are transcribed by the genome. However, they are not translated into proteins but rather control translation of coding mRNA. Particularly in the brain, numerous different microRNAs are expressed in a cell type specific fashion both during development and in adulthood. Aberrant microRNA expression has been implicated in several human diseases including CNS diseases. The aim of this review is to emphasize their role in the development of the brain and their function. In addition, we highlight recent findings on the evolution of mammalian microRNAs and their effect on steroid signaling in the brain. PMID- 19683018 TI - The Bacillus anthracis spore. AB - In response to starvation, Bacillus anthracis can form a specialized cell type called the spore, which is the infectious particle for the disease anthrax. The spore is largely metabolically inactive and can resist a wide range of stresses found in nature. In spite of its dormancy, the spore can sense the presence of nutrient and rapidly return to vegetative growth. These properties help the spore to persist for long periods of time in the environment, survive host defenses after entering the body, and cause disease when the correct location in the host is reached. The anatomy of the spore is unique among bacteria, being comprised of a series of specialized concentric shells, each of which provides specific critical functions. Surrounding the spore core (which houses the chromosome) is a peptidoglycan layer important for spore dormancy, a protein shell that resists a variety of toxic molecules, and finally an exterior protein and glycoprotein layer that, among other functions, mediates interactions with surfaces, including those encountered by the spore within the host. Detailed molecular analysis of these shells has shed considerable light on how each layer determines specific spore properties. Future work, especially on the outermost spore layer, is likely to advance therapeutics, methods for spore decontamination and other critical biodefense technologies. PMID- 19683020 TI - Cytoskeletal alterations in rat hippocampus following chronic unpredictable mild stress and re-exposure to acute and chronic unpredictable mild stress. AB - The intrinsic dynamic instability of the cytoskeletal microtubular system is essential for neuronal development and organization. The modulation of microtubule dynamics depends on the phosphorylation of neuronal microtubule associated proteins (MAPs). Chronic unpredicted mild stress (CUMS) affects hippocampal structure and function in the rat. The aim of the present work was to investigate the possible alteration of cytoskeleton in the hippocampus of rats exposed to CUMS and re-exposed to CUMS to mimic depression and the recurrence of depression of human. We investigated the effects of CUMS, fluoxetine and re exposure to CUMS on alpha-tubulin isoforms associated with microtubule dynamics, MAP-2 and phospho-MAP-2 in the hippocampus of rats. Our results showed that rats submitted to CUMS once showed a significant reduction in locomotion and sucrose preference which indicate a state of anhedonia. These behavioral alterations were accompanied by specific alterations in hippocampal alpha-tubulin isoforms and phospho-MAP-2 expression, indicating less microtubule dynamics and the possible mechanism. Treatment of fluoxetine could reverse CUMS-induced impairment. Moreover, there were more dramatically changes in behaviors, alpha-tubulin isoforms and phospho-MAP-2 of rats re-exposed to CUMS compared to the rats exposed to CUMS once. These findings provide evidence that rats exposed to CUMS and re-exposed to CUMS showed impairment of microtubule dynamics accompanied with the decreased level of phospho-MAP-2, providing insight into the role of cytoskeleton in the depression and recurrent of depression. PMID- 19683021 TI - Involvement of the serotonergic system in the anxiolytic-like effect caused by m trifluoromethyl-diphenyl diselenide in mice. AB - The organoselenium compound diphenyl diselenide (PhSe)(2) has shown interesting antioxidant and neuroprotective activities. On the other hand, this compound has also presented some toxic effects. m-Trifluoromethyl-diphenyl diselenide (m-CF(3) C(6)H(4)Se)(2), a structural analog of (PhSe)(2), has proven to be antipsychotic and antioxidant in mice. The present study was designed to investigate the anxiolytic-like effect of (m-CF(3)-C(6)H(4)Se)(2) in female mice, employing light/dark box and elevated plus-maze (EPM) tests. The involvement of 5 hydroxytryptamine (5-HT) receptors and monoamine oxidase (MAO) activity in the anxiolytic-like effect was also evaluated. (m-CF(3)-C(6)H(4)Se)(2) (0.1, 10 and 100 mg/kg, p.o.) did not affect locomotor activity as evaluated in the open-field test (OFT). (m-CF(3)-C(6)H(4)Se)(2) at the dose of 100 mg/kg produced an anxiolytic-like action, both in light-dark box and the EPM tests. To evaluate the role of 5-HT receptors in the anxiolytic-like effect of (m-CF(3)-C(6)H(4)Se)(2), a selective 5-HT(1A) receptor antagonist, WAY100635 (0.1 mg/kg, s.c.), a non selective 5-HT(2A/2C) receptor antagonist, ritanserin (2 mg/kg, i.p.) and a selective 5-HT(3) receptor antagonist, ondansetron (0.1 mg/kg, i.p.) were used. All the antagonists used were able to abolish the anxiolytic-like effect of (m CF(3)-C(6)H(4)Se)(2). (m-CF(3)-C(6)H(4)Se)(2), at the dose of 100 mg/kg, inhibited the MAO-A activity in mice brain. Taken together these data demonstrated that the anxiolytic-like effect caused by (m-CF(3)-C(6)H(4)Se)(2) seems to be mediated by the involvement of the serotonergic system. PMID- 19683022 TI - Lack of association of NKX2-3, IRGM, and ATG16L1 inflammatory bowel disease susceptibility variants with celiac disease. AB - Evidence about the presence of susceptibility factors shared among different autoimmune diseases is increasing. Based on this idea, NKX2-3, ATG16L1, and IRGM which are well-established inflammatory bowel disease risk factors, could be new celiac disease (CD) candidate genes. NKX2-3 encodes a transcription factor that in mice seems to be involved in gut development. The ATG16L1 and IRGM genes act in autophagy, a process related to innate and adaptive immunity. We aimed to study the implication of five polymorphisms in these genes in CD susceptibility: rs10883365 and rs888208 in the NKX2-3 gene, rs2241880 in ATG16L1, and rs10065172 and rs4958847 in IRGM. Association studies were performed using 725 Spanish CD patients and 956 ethnically matched healthy controls, as well as 309 parent-child trios. Genetic frequencies were compared with the chi(2) test and the familial study used the transmission disequilibrium test. Differences between CD patients and controls did not reach significance when genotypic and allelic frequencies were compared. No differential transmission of alleles or haplotypes from heterozygous parents to affected children was observed in the familial study. In conclusion, no evidence of association with CD has been reported for the Crohn's disease susceptibility polymorphisms studied in the NKX2-3, ATG16L1, and IRGM genes. PMID- 19683023 TI - Estimation of high-resolution HLA-A, -B, -C, -DRB1 allele and haplotype frequencies based on 8862 German stem cell donors and implications for strategic donor registry planning. AB - Human leukocyte antigen (HLA) haplotype frequency distributions in specific populations can be applied to optimize both individual stem cell donor searches and donor registry planning. We present allele and haplotype frequencies derived from a data set of 8862 German stem cell donors who were typed at high resolution for the HLA-A, HLA-B, HLA-C, and HLA-DRB1 genes upon registration. Calculated haplotype frequencies were used to estimate the probability p to find matching donors subject to donor registry size n. The impact of various matching standards on p(n) was analyzed. When high-resolution matching for HLA-A, HLA-B, HLA-C, and HLA-DRB1 is required, p(1,000,000) is 0.678. The corresponding value for n = 7,000,000 is 0.859. In a scenario with low-resolution matching and no consideration of HLA-C, p(1,000,000) is 0.863 and thus larger than p(7,000,000) in the scenario with stricter matching requirements. As recent findings support the importance of high-resolution matching of HLA-A, HLA-B, HLA-C, and HLA-DRB1 for outcomes of hematopoietic stem cell transplantation, our results are highly relevant for strategic planning and resource allocation of donor centers and registries. PMID- 19683019 TI - Anti-relapse medications: preclinical models for drug addiction treatment. AB - Addiction is a chronic relapsing brain disease and treatment of relapse to drug seeking is considered the most challenging part of treating addictive disorders. Relapse can be modeled in laboratory animals using reinstatement paradigms, whereby behavioral responding for a drug is extinguished and then reinstated by different trigger factors, such as environmental cues or stress. In this review, we first describe currently used animal models of relapse, different relapse triggering factors, and the validity of this model to assess relapse in humans. We further summarize the growing body of pharmacological interventions that have shown some promise in treating relapse to psychostimulant addiction. Moreover, we present an overview on the drugs tested in cocaine or methamphetamine addicts and examine the overlap of existing preclinical and clinical data. Finally, based on recent advances in our understanding of the neurobiology of relapse and published preclinical data, we highlight the most promising areas for future anti-relapse medication development. PMID- 19683024 TI - Association of human leukocyte antigens with nasopharyngeal carcinoma in high risk multiplex families in Taiwan. AB - An association between specific human leukocyte antigens (HLA) alleles and nasopharyngeal carcinoma (NPC) has been reported for sporadic NPC, but studies of familial NPC are lacking. We evaluated this association with familial NPC in a study of 301 NPC cases and 1010 family and community controls from Taiwan. Class I HLA alleles were characterized using a sequence-based typing protocol. Allele frequencies between case and control groups were compared by chi(2) or exact tests. For alleles associated with NPC, odds ratios (OR) and 95% confidence intervals (CI) were calculated. Similar allelic frequency distribution and HLA associations were found as those previously reported for sporadic NPC: protective effect for HLA-A*1101 and increased risk for HLA-A*0207, HLA-A*3303, HLA-B*3802, and HLA-B*5801. Overall, the magnitude of observed associations was weakest when cases were compared with sibling controls and strongest when compared with unrelated community controls. Evaluating the joint effect of HLA-A*0207 and HLA B*4601, individuals who were carriers of HLA-A*0207 with or without the presence of HLA-B*4601 had a 1.9-fold (95% CI = 1.0-3.4) and 2.1-fold (95% CI = 0.83-5.3) risk of NPC, respectively. Conversely, carriers of HLA-B*4601 in the absence of HLA-A*0207 had a 50% reduction in NPC risk (95% CI = 0.27-0.93). Comparable findings from our family study and those from previous sporadic studies were found with the notable exception of a lack of positive association between HLA B*4601 and familial NPC in the absence of HLA-A*0207. This finding requires replication in larger studies. PMID- 19683026 TI - The symptomatic profile of panic disorder is shaped by the 5-HTTLPR polymorphism. AB - The short allele of a functional polymorphism (5-HTTLPR) in the serotonin transporter (5-HTT) promoter is associated with reduced serotonin transporter expression, lower in vivo 5-HTT binding, higher neuroticism and amygdala reactivity as well as facilitated fear conditioning and is a candidate variant for panic disorder. However, case-control studies have consistently failed to show an association between 5-HTTLPR and panic disorder. As psychiatric disorders are broadly defined phenotypes merging different symptoms to syndromes, they may not be very well suited for genetic association studies. An alternative approach is to measure symptoms along continuous symptom dimensions which may more appropriately reflect their biological underpinnings and may reveal subpopulations within clinical diagnostic groups. We recorded the symptomatic profile in 73 in panic disorder patients using observer-rated instruments (Panic Disorder Severity Scale, PDSS; Montgomery-Asberg Depression Rating Scale, MADRS) and hypothesized more severe symptoms in patients carrying the 5-HTTLPR s-allele. We observed a strong association between bi- and triallelic 5-HTTLPR polymorphisms and the symptomatic profile. Carriers of the 5-HTTLPR s-allele suffered from most severe panic and depressive symptoms. Our data highlight the importance of defining appropriate phenotypes for psychiatric genetic studies and demonstrate that the 5-HTTLPR genotype may be related to the symptomatic profiles rather than to the vulnerability to develop panic disorder. PMID- 19683027 TI - Protective effects of saponins from the root of Platycodon grandiflorum against fatty liver in chronic ethanol feeding via the activation of AMP-dependent protein kinase. AB - Fatty liver and steatosis induced by alcohol is the earliest and most common response of the liver to alcohol and may be a precursor of more severe forms of liver injury. However, the mechanism of liver injury and deposition of fatty liver due to alcohol is complex. The protective effects of saponins from the root of Platycodon grandiflorum (Changkil saponins: CKS) against ethanol-induced liver injury in an enteral alcohol feeding model was investigated. Male Sprague-Dawley rats were given control diets or ethanol-containing diets enterally for 4 weeks. Treatment with CKS for 2 weeks significantly prevented the alcohol-induced increase in serum alanine aminotransferase and aspartate aminotransferase activities or decrease in serum albumin levels. Alcohol elevated the hepatic triglyceride content and induced cytochrome P450 2E1 (CYP2E1) expression. CKS treatment reduced CYP2E1 expression and hepatic triglyceride accumulation and prevented alcoholic liver steatosis. Chronic alcohol feeding decreased AMP activated protein kinase-alpha (AMPKalpha) phosphorylation, which was restored by CKS treatment. Recovery of AMPKalpha phosphorylation by CKS was also followed by an increase in acetyl-CoA carboxylase phosphorylation. Our study suggests that CKS is a promising agent for preventing or treating human alcoholic fatty liver disease. PMID- 19683028 TI - Zinc bioaccumulation in a terrestrial invertebrate fed a diet treated with particulate ZnO or ZnCl2 solution. AB - A number of reports on potential toxicity of nanoparticles are available, but there is still a lack of knowledge concerning bioaccumulation. The aim of this work was to investigate how different sources of zinc, such as uncoated and unmodified ZnO nanoparticles, ZnCl(2) in solution, and macropowder ZnO influence the bioaccumulation of this metal in the terrestrial isopod Porcellio scaber. After exposure to different sources of Zn in the diet, the amount of assimilated Zn in whole body, the efficiency of zinc assimilation, and bioaccumulation factors (BAFs) were assessed. The bioaccumulation potential of Zn was found to be the same regardless of Zn source. The amount of assimilated Zn and BAF were dose dependent, and Zn assimilation efficiency was independent of exposure concentrations. The Zn assimilation capacity was found to be up to 16% of ingested Zn. It is known that as much as approximately 20% of Zn can be accreted from ZnO particles by dissolution. We conclude that bioaccumulation of Zn in isopods exposed to particulate ZnO depends most probably on Zn dissolution from ZnO particles and not on bioaccumulation of particulate ZnO. PMID- 19683025 TI - Intrinsic neuronal plasticity in the juxtacapsular nucleus of the bed nuclei of the stria terminalis (jcBNST). AB - The juxtacapsular nucleus of the anterior division of the BNST (jcBNST) receives robust glutamatergic projections from the basolateral nucleus of the amygdala (BLA), the postpiriform transition area, and the insular cortex as well as dopamine (DA) inputs from the midbrain. In turn the jcBNST sends GABAergic projections to the medial division of the central nucleus of the amygdala (CEAm) as well as other brain regions. We recently described a form of long-term potentiation of the intrinsic excitability (LTP-IE) of neurons of the juxtacapsular nucleus of BNST (jcBNST) in response to high-frequency stimulation (HFS) of the stria terminalis that was impaired during protracted withdrawal from alcohol, cocaine, and heroin and in rats chronically treated with corticotropin releasing factor (CRF) intracerebroventricularly. Here we show that DAergic neurotransmission is required for the induction of LTP-IE of jcBNST neurons through dopamine (DA) D1 receptors. Thus, activation of the central CRF stress system and altered DAergic neurotransmission during protracted withdrawal from alcohol and drugs of abuse may contribute to the disruption of LTP-IE in the jcBNST. Impairment of this form of intrinsic neuronal plasticity in the jcBNST could result in inadequate neuronal integration and reduced inhibition of the CEA, contributing to the negative affective state that characterizes protracted abstinence in post-dependent individuals. These results provide a novel neurobiological target for vulnerability to alcohol and drug dependence. PMID- 19683029 TI - Action potential changes associated with the inhibitory effects on voltage-gated sodium current of hippocampal CA1 neurons by silver nanoparticles. AB - Nano-sized materials are now being used in medicine, biotechnology, energy, and environmental technology. Although a wide and growing number of applications for nanomaterials exist, there are limited studies available on toxicity of nanoparticles for their human risk and environmental assessment. The aim of this study was to investigate the effects of silver nanoparticles (nano-Ag) on voltage activated sodium currents in hippocampal CA1 neurons. Nano-Ag was tested at increasing concentrations (10(-6), 5 x 10(-6), 10(-5) g/ml). The research results showed that only nano-Ag (10(-5) g/ml) reduced the amplitude of voltage-gated sodium current (I(Na)). The nano-Ag particles produced a hyperpolarizing shift in the activation-voltage curve of I(Na) and also delayed the recovery of I(Na) from inactivation. Action potential properties and the pattern of repetitive firing were examined using whole cell current-clamp recordings. Peak amplitude and overshoot of the evoked single action potential were decreased and half-width was increased in the present of the 10(-5) g/ml nano-Ag solution, and the firing rate of repetitive firing had no change. The results suggest that nano-Ag may alter the action potential of hippocampal CA1 neurons by depressing voltage-gated sodium current. PMID- 19683030 TI - Protective effects of thiopronin against isoniazid-induced hepatotoxicity in rats. AB - Isoniazid is a widely used drug for the treatment of tuberculosis, but hepatotoxicity is a major concern during treatment. Thiopronin contains an SH group and is generally considered an antioxidant. The aim of the present study was to investigate the effects of thiopronin during liver injury and DNA damage induced by isoniazid. Rats were injected daily with isoniazid (100 mg/kg, i.p.) for 21 days with or without thiopronin co-administration (60 mg/kg, i.p.) from day 11 to day 21. The influence of thiopronin on isoniazid-induced DNA oxidative damage was analyzed in precision-cut rat liver slices by HPLC-MS/MS. Thiopronin prevented isoniazid-induced hepatotoxicity, indicated by both diagnostic indicators of liver damage (alanine aminotransferase and aspartate aminotransferase) and histopathological analysis. In vivo, thiopronin significantly inhibited isoniazid-induced CYP2E1 activity as assessed by both chlorzoxazone hydroxylase and aniline hydroxylase (p<0.001). Thiopronin concentration-dependently inhibited CYP2E1-dependent aniline hydroxylation, and the Dixon plots suggest that thiopronin is a competitive inhibitor of CYP2E1. Thiopronin markedly attenuated isoniazid-induced inhibition of the detoxification system through cytosolic glutathione S-transferases (GSTs), including mu GST and alpha GST. In precision-cut liver slices, the free radical scavenging activity of thiopronin reduced the generation of DNA adducts induced by isoniazid (p<0.05). Altogether, these results suggest that thiopronin exerts its hepatoprotective activity against isoniazid-induced hepatotoxicity by inhibiting the production of free radicals in addition to its role as a scavenger. Thiopronin may reduce free radical generation via inhibition of hepatic CYP2E1 and increase the removal of free radicals directly or through the induction of cytosolic GSTs. PMID- 19683031 TI - Cell based approaches for evaluation of drug-induced liver injury. AB - An improved understanding of mechanisms that underlie drug-induced liver injury (DILI) is required to enable design of drugs that have minimal potential to cause this adverse reaction in man. Available evidence suggests DILI arises in susceptible patients because of an imbalance between chemical insults (which are an inherent property of certain drugs and/or their metabolites) and the ability of the liver to mount compensatory/adaptive responses. In vivo safety testing in pre-clinical species ensures that drugs which enter clinical trials do not cause reproducible and dose-dependent liver injury in man, but is of limited value for exploration of underlying mechanisms and does not assess potential to cause rare idiosyncratic DILI. This review highlights the value that can be gained from in vitro studies using cultured hepatocytes and also hepatocyte-derived cell lines transfected with individual human cytochrome P450 (CYP450) isoforms. We have evaluated a range of mechanisms and endpoints (cell necrosis, mitochondrial injury, inhibition of biliary transporters and metabolite-mediated toxicity) using these model systems. Our data indicate that multiple mechanisms are likely to be involved in development of idiosyncratic DILI in man caused by numerous drugs, e.g. the anticonvulsant chlorpromazine. PMID- 19683032 TI - The first alpha-1,3-glucosidase from bacterial origin belonging to glycoside hydrolase family 31. AB - Genome analysis of Lactobacillus johnsonii NCC533 has been recently completed. One of its annotated genes, lj0569, encodes the protein having the conserved domain of glycoside hydrolase family 31. Its homolog gene (ljag31) in L. johnsonii NBRC13952 was cloned and expressed using an Escherichia coli expression system, resulting in poor production of recombinant LJAG31 protein due to inclusion body formation. Production of soluble recombinant LJAG31 was improved with high concentration of NaCl in medium, possible endogenous chaperone induction by benzyl alcohol, and over-expression of GroES-GroEL chaperones. Recombinant LJAG31 was an alpha-glucosidase with broad substrate specificity toward both homogeneous and heterogeneous substrates. This enzyme displayed higher specificity (in terms of k(cat)/K(m)) toward nigerose, maltulose, and kojibiose than other natural substrates having an alpha-glucosidic linkage at the non-reducing end, which suggests that these sugars are candidates for prebiotics contributing to the growth of L. johnsonii. To our knowledge, LJAG31 is the first bacterial alpha-1,3-glucosidase to be characterized with a high k(cat)/K(m) value for nigerose [alpha-d-Glcp-(1 --> 3)-d-Glcp]. Transglucosylation of 4-nitrophenyl alpha-d-glucopyranoside produced two 4-nitrophenyl disaccharides (4-nitrophenyl alpha-nigeroside and 4-nitrophenyl alpha-isomaltoside). These hydrolysis and transglucosylation properties of LJAG31 are different from those of mold (Acremonium implicatum) alpha-1,3-glucosidase of glycoside hydrolase family 31. PMID- 19683033 TI - From care to cure. PMID- 19683035 TI - Cephalic index and perceived dog trainability. AB - People rank breeds of dogs for trainability despite a lack of evidence of breed differences in underlying behaviour. Instead of using behavioural information, people may use dog morphology to determine the trainability of breeds. Dogs are categorized as dolichocephalic, mesocephalic, or brachycephalic based on cephalic index, a ratio between skull width and length. Dolichocephalic breeds are anatomically more specialized for running and brachycephalic breeds are more specialized for fighting. Dog breeds rated as highly trainable are instead mesocephalic, morphological generalists. Looking trainable in dogs may reflect differences in physical morphology. PMID- 19683034 TI - Axonopathy and cytoskeletal disruption in degenerative diseases of the central nervous system. AB - There has been growing interest in the axon as the initial focus of pathological change in a number of neurodegenerative diseases of the central nervous system. This review concentrates on three major neurodegenerative conditions--amyotrophic lateral sclerosis, multiple sclerosis and Alzheimer's disease--with emphasis on key cellular changes that may underlie early axonal dysfunction and pathology and, potentially, the degeneration of neurons. In particular, this review will address recent data that indicate that the main pathological stimuli for these conditions, though often not definitively determined, result in an initial perturbation of the axon and its cytoskeleton, which then results in slow neuronal degeneration and loss of connectivity. The identification of a degenerative process initiated in the axon may provide new therapeutic targets for early intervention to inhibit the grim outcomes related to the progression of these diseases. PMID- 19683036 TI - Choose-long errors at delays shorter than the training delay persist when enhanced between-trial summation of duration memories are precluded in pigeons. AB - Pigeons were trained to match 2- and 10-s durations of houselight to red and green comparisons. Following acquisition with a 0-s baseline delay, they were tested with delays of 0, 10 and 20s. There was a strong tendency to choose the short-associated comparison stimulus at both the 10- and 20-s delays (i.e., a choose-short effect) and no bias at the 0-s delay. This test was repeated after baseline training with a constant 10-s delay. As in the first delay test, a choose-short effect was obtained at the 20-s delay. In contrast to the first test, no bias was obtained at the 10-s delay and a strong tendency to choose the long-associated comparison (i.e., a choose-long effect) was obtained at the 0-s delay. Importantly, the choose-long effect was obtained under conditions which insured that the temporal spacing between a 0-s delay trial and the preceding trial was equal to that in training. These results are inconsistent with the temporal summation account of the choose-long effect and are most readily interpreted within a perspective emphasizing the subjective shortening of temporal memories over time. PMID- 19683037 TI - Rats' use of geometric, featural and orientation cues to locate a hidden goal. AB - Over the past 20 years, a great deal of research has examined how different animals can use the geometric properties of the environment to determine their heading. Less well studied is how rats use the geometric properties of an environment to navigate, or determine the location, when it is not necessary to establish heading. Specifically, it is unclear to what extent rats still rely on geometric cues when they are not disoriented. In the current study, rats were trained to find food in one corner of a rectangular environment under either oriented or disoriented conditions. Probe tests placed geometric, featural and orientation cues in conflict. Results showed that featural cues exerted little control over the rats' search preferences. All rats, whether trained while oriented or trained while disoriented, used geometric cues when these were the only cues available. Rats trained in the disoriented condition preferred geometric cues to orientation cues, whereas rats trained in the oriented condition showed more equal preference for orientation and geometric cues. PMID- 19683039 TI - Structural, evolutionary and genetic analysis of the histidine biosynthetic "core" in the genus Burkholderia. AB - In this work a detailed analysis of the structure, the expression and the organization of his genes belonging to the core of histidine biosynthesis (hisBHAF) in 40 newly determined and 13 available sequences of Burkholderia strains was carried out. Data obtained revealed a strong conservation of the structure and organization of these genes through the entire genus. The phylogenetic analysis showed the monophyletic origin of this gene cluster and indicated that it did not undergo horizontal gene transfer events. The analysis of the intergenic regions, based on the substitution rate, entropy plot and bendability suggested the existence of a putative transcription promoter upstream of hisB, that was supported by the genetic analysis that showed that this cluster was able to complement Escherichia colihisA, hisB, and hisF mutations. Moreover, a preliminary transcriptional analysis and the analysis of microarray data revealed that the expression of the his core was constitutive. These findings are in agreement with the fact that the entire Burkholderiahis operon is heterogeneous, in that it contains "alien" genes apparently not involved in histidine biosynthesis. Besides, they also support the idea that the proteobacterial his operon was piece-wisely assembled, i.e. through accretion of smaller units containing only some of the genes (eventually together with their own promoters) involved in this biosynthetic route. The correlation existing between the structure, organization and regulation of his "core" genes and the function(s) they perform in cellular metabolism is discussed. PMID- 19683040 TI - ISBM-7: Biological Monitoring in a Globalized World. Preface. PMID- 19683038 TI - Functional characterization of PAS and HES family bHLH transcription factors during the metamorphosis of the red flour beetle, Tribolium castaneum. AB - The basic helix-loop-helix transcription factors are present in animals, plants and fungi and play important roles in the control of cellular proliferation, tissue differentiation, development and detoxification. Although insect genomes contain more than 50 helix-loop-helix transcription factors, the functions of only a few are known. RNAi has become a widely used tool to knock-down the expression to analyze the function of genes. As RNAi works well in Tribolium castaneum, we utilized this insect and RNAi to determine functions of 19 bHLH transcription factors belonging to PAS and HES families during the larval stages of the red flour beetle, T. castaneum. We searched the genome sequence of T. castaneum and identified 53 bHLH genes. Phylogenetic analyses classified these 53 genes into ten families; PAS, HES, Myc/USF, Hand, Mesp, Shout, p48, NeuroD/Neurogenin, Atonal and AS-C. In RNAi studies, knocking-down the expression of seven members of the PAS and HES families affected the growth and development of T. castaneum. An inability to grow to reach critical weight to undergo metamorphosis, failure to complete larval-pupal or pupal-adult ecdysis and abnormal wing development are among the most common phenotypes observed in RNAi insects. Among the bHLH transcription factors studied, the steroid receptor coactivator (SRC) showed the most severe phenotypes. Knock-down in the expression of the gene coding for SRC caused growth arrest by affecting the regulation of lipid metabolism. These studies demonstrate the power of RNAi for functional characterization of members of the multigene families in this model insect. PMID- 19683041 TI - Chronic exposure to MDMA (ecstasy) increases DNA damage in sperm and alters testes histopathology in male rats. AB - 3,4-Methylenedioxymethamphetamine (MDMA or "ecstasy") is consumed mainly by young population. For this reason, it is especially relevant to take into consideration the effects on the reproductive system. The influence of MDMA on the fertility and reproduction of the male rat was assessed in this study. MDMA was administered subcutaneously at 0 mg/kg (control), 0.5 mg/kg, 5 mg/kg and 10 mg/kg to SD male rats once a day, 3 consecutive days a week during 12 weeks, simulating human weekend associated consumption. Hormonal, haematological, biochemical, histological, genotoxicological and testicular and sperm parameters were evaluated in half of the rats. The remaining animals were mated with untreated sexually receptive females to evaluate the mating and pregnancy rates. A significantly higher incidence of DNA damage in Comet Test in sperm, tubular degeneration and interstitial oedema in testes was found. At all doses tested, sperm motility, morphology, mating and pregnancy rates, and number of implantation sites were not affected. This study fills the existing gap of knowledge about the chronic effects of MDMA in reproductive function using a realistic experimental design. Taking into account the higher sensitivity of human males, some concerns about the effects on the reproductive health still remain. PMID- 19683042 TI - Effects of ultraviolet radiation on the kinetics of in vitro percutaneous absorption of lavender oil. AB - The purpose of the present study is to investigate the influence of ultraviolet radiation on the rat skin absorption of lavender essential oil. The pure oil was extracted from Lavandula angustifolia by steam distillation. The chemical composition of lavender oil showed that terpenes are major compounds. In vitro, the essential oil was applied onto the rat skin. The amount of the compounds was determined using gas chromatography. Similarly, the amount of these compounds was analyzed for the skin exposed to ultraviolet radiation (UVAI) after 4, 8, 12 and 24 h. Our study demonstrated that the penetration profiles showed a cycle of charge-discharge (4 h/4 h, respectively). Our data point to the presence of reversible change in stratum corneum behavior. Interestingly, the ultraviolet radiation altered the cycle (charge-discharge) for terpenes (low lipophilicity) and increased the charge time. However, for terpenes (high lipophilicity), the ultraviolet radiation decreased the charge amplitude. PMID- 19683043 TI - Clinical trial to compare the effectiveness of two concentrations of the Ageratina pichinchensis extract in the topical treatment of onychomycosis. AB - The plant species Ageratina pichinchensis has been used, for many years, in Mexican traditional medicine for the treatment of superficial mycosis. AIM OF THE STUDY: This study compared the therapeutic effectiveness and tolerability of two concentrations of the standardized extract from Ageratina pichinchensis (12.6 and 16.8%) on patients with clinical and mycological diagnosis of mild and moderate onychomycosis. MATERIALS AND METHODS: Two identical phytopharmaceuticals (containing the standardized extract from Ageratina pichinchensis) in nail lacquer solution for topical administration were evaluated in a double-blind clinical trial. Treatments were administered for 6 months to patients distributed in two groups. RESULTS AND DISCUSSION: Of 122 patients who agreed to participate in the study, 103 (84.4%) concluded the treatment. The therapeutic effectiveness exhibited by the 12.6% Ageratina pichinchensis extract was 67.2%, while that of the 16.8% Ageratina pichinchensis extract was 79.1%. Regarding clinical evolution, analysis of results at the end of treatment evidenced that the 16.8% concentration possesses higher therapeutic effectiveness with a significant statistical difference (p=0.010). No treatment produced side effects. CONCLUSION: Both concentrations of phytopharmaceuticals possess high rates of effectiveness on patients with mild and moderate onychomycosis, and the formulation with a 16.8% concentration possesses higher effectiveness. PMID- 19683044 TI - Anti-inflammatory activity of ethanol extract from Geranium sibiricum Linne. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Geranium sibiricum (Geraniaceae) Linne (GSL) is used to heal various disorders of the diarrhea and the intestinal inflammation as an herbal agent in East Asia. AIMS OF THE STUDY: The purpose of the present study is to determine whether the ethanol (EtOH) extract of GSL regulates the inflammatory reaction stimulated by phorbol-12-myristate 13-acetate plus calcium ionophore A23187 (PMACI) in human mast cells (HMC-1). MATERIALS AND METHODS: Western blot was used for activation of mitogen activated protein kinase (MAPK), transcription factors, induced nitric oxide synthase (iNOS), and cyclooxygenase (COX)-2 proteins. EMSA was for DNA binding activity. RT-PCR was used for gene expression. RESULTS: EtOH extract of GSL (EGS) inhibits the expression of extracellular signal-regulated kinase (ERK), one of a MAPK, nuclear transcription factors involving nuclear factor (NF)-kappaB and Activator protein (AP)-1, COX-2 and iNOS. The results indicated that EGS decreased gene expression of interleukin (IL)-1beta and COX-2 in PMACI stimulated HMC-1 cells. CONCLUSION: Hence, we speculate that EGS can use as a potent anti-inflammatory agent for inflammatory allergic diseases. PMID- 19683045 TI - Quality and safety of Chinese herbal medicines guided by a systems biology perspective. AB - Chinese herbal medicines, often referred as Chinese materia medica (CMM), are comprised of a complex multicomponent nature. The activities are aimed at the system level via interactions with a multitude of targets in the human body. This review aims at the toxicity aspects of CMM and its preparations at the different steps of production; harvesting, processing and the final formulation. The historic perspective and today's issues of the safety of CMM are introduced briefly, followed by the descriptions of the toxic CMM in the current Chinese Pharmacopoeia (2005). Subsequently, several aspects of safety are illustrated using a typical example of a toxic CMM, Aconitum roots, and some recent findings of our own research are included to illustrate that proper processing and multi herbs formulation can reduce the level of toxic components. This also explains that in CMM, some herbs, such as Aconitum, Ephedra species are never used as single herb for intervention and that aconite is only used when it is processed and in combination with specific matched other herbs. The formulation principle of multi-herbs intervention strategy is a systems approach for the treatment and prevention of disease. In this light, the role of systems toxicology in the safety and quality of Chinese herbal medicine is proposed as a promising method. Moreover the principles of practiced-based and evidence-based research are discussed from a symbiotic perspective. PMID- 19683046 TI - Effect of Solanum torvum on blood pressure and metabolic alterations in fructose hypertensive rats. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Solanum torvum (Solanaceae) is a plant used in Cameroon ethnomedicine for the treatment of hypertension. AIM OF THE STUDY: The present study was aimed to determine the effect of ethanolic extract of Solanum torvum (100 and 300 mg/kg; p.o. for 6 weeks) on systolic blood pressure (SBP), vascular reactivity, serum glucose, triglycerides, cholesterol, insulin and uric acid in fructose-induced hypertension. MATERIALS AND METHODS: The effect of ethanolic extract of Solanum torvum (100 and 300 mg/kg; p.o. for 6 weeks) on fructose (10%) induced rise in blood pressure was tested by invasive and non invasive measurements and the biochemical parameters were studied by using standard kits. RESULTS: Ethanolic extract of Solanum torvum reduced systolic blood pressure, vascular reactivity changes to catecholamines and reversed the metabolic alterations induced by fructose. The cumulative concentration response curve (CCRC) of Angiotensin II (Ang II) using isolated strip of ascending colon was shifted towards right in rats treated with ethanolic extract of Solanum torvum. CONCLUSIONS: In conclusion, ethanolic extract of Solanum torvum could prevent the development of high blood pressure induced by a diet rich in fructose probably by reversing the metabolic alterations induced by fructose. PMID- 19683047 TI - Evidence that the major metabolites accumulating in hyperornithinemia hyperammonemia-homocitrullinuria syndrome induce oxidative stress in brain of young rats. AB - Ornithine and homocitrulline are the major metabolites accumulating in hyperornithinemia-hyperammonemia-homocitrullinuria syndrome, a genetic disorder characterized by neurological regression whose pathogenesis is still not understood. The present work investigated the in vitro effects of ornithine and homocitrulline on important parameters of oxidative stress in cerebral cortex from young rats. Ornithine significantly increased chemiluminescence and thiobarbituric acid-reactive substances levels, indicators of lipid peroxidation, while homocitrulline only augmented chemiluminescence values. Furthermore, ornithine-induced increase of thiobarbituric acid-reactive substances levels was attenuated (melatonin and reduced glutathione) or totally prevented (alpha tocopherol) by free radical scavengers, suggesting that reactive species were involved in the lipid oxidative damage. We also observed that ornithine and homocitrulline significantly decreased the tissue antioxidant defenses, determined by reduced glutathione concentrations, the major non-enzymatic antioxidant defense found in the brain. Homocitrulline reduction of glutathione levels was completely prevented by melatonin and alpha-tocopherol, whereas ornithine-induced decrease of glutathione levels was only attenuated by these free radical scavengers. Ornithine and homocitrulline also induced protein oxidative damage, increasing carbonyl formation and sulfhydryl oxidation. In contrast, these amino acids did not affect nitric oxide production, indicating that nitrogen reactive species were not implicated in the lipid and oxidative damage provoked by ornithine and homocitrulline. Therefore, it is presumed that the major metabolites accumulating in hyperornithinemia-hyperammonemia homocitrullinuria syndrome elicit oxidative stress and that this pathomechanism may possibly be involved in the brain damage found in patients affected by this disorder. PMID- 19683048 TI - MexT modulates virulence determinants in Pseudomonas aeruginosa independent of the MexEF-OprN efflux pump. AB - In the human pathogen Pseudomonas aeruginosa, the LysR-family regulator MexT modulates the induction of the tripartite MexEF-OprN resistance nodulation division multi-drug efflux system resulting in increased resistance to diverse antibiotics. The MexEF-OprN system is normally quiescent in wild-type cells, but is highly induced in nfxC-type phenotypic mutants in a MexT dependent manner. In addition to antibiotic resistance, induction of mexEF-oprN in nfxC-type mutants has been linked to reduced levels of homoserine lactone-dependent virulence traits, including pyocyanin, elastase, rhamnolipids and PQS and to reduced expression of type three secretion effector proteins. In this study, MexT is overexpressed in wild-type PAO1 and an isogenic mexEF deletion mutant to determine if MexT regulates diverse virulence phenotypes dependent or independent of MexEF-OprN. It is shown that MexT regulates type three secretion, pyocyanin production and early surface attachment independent of MexEF-OprN. In contrast, MexT modulation of the expression of the virulence genes rhlA, lasB and hcnB is dependent on MexEF-OprN, which apparently mediates these effects via efflux of cell-signaling intermediates. The data presented demonstrates that MexT may play a more global role in modulating P. aeruginosa virulence than previously reported and suggests that MexT regulates diverse targets that mediate phenotypic alterations independent of MexEF-OprN. PMID- 19683049 TI - Inhibition of heat shock protein expression by Helicobacter pylori. AB - Heat shock proteins (HSPs) are primarily known as molecular chaperones that are induced by cell stress and prevent protein aggregation and facilitate folding. Recent evidence suggests that exposure of cells to microbial pathogens can also induce HSPs, which then modulate both innate and adaptive immune responses. Paradoxically, Helicobacter pylori has been found to decrease expression of HSPs. We sought to investigate this phenomenon further and to examine the role of different H. pylori strains and recognized virulence factors in cell culture and in the mouse model. Co-culture of H. pylori with two gastric carcinoma cell lines reduced expression of HSP70 and, to a lesser extent, HSP60. Down modulation of HSPs was not dependent on the presence of the vacuolating cytotoxin (VacA) or the cag pathogenicity island (cag PAI). C57BL/6 mice infected with a human H. pylori strain also demonstrated reduced expression of HSP70, HSP8, and heat shock factor 1 (HSF-1), a transcriptional activator of HSP70. In contrast, the bacterial pathogen, S. Typhimurium up-regulated HSP expression. Since HSPs are thought to function as danger signals during microbial infection, H. pylori down-regulation of HSPs may be a mechanism of immune evasion that promotes chronic infection. PMID- 19683050 TI - Bezafibrate induces myotoxicity in human rhabdomyosarcoma cells via peroxisome proliferator-activated receptor alpha signaling. AB - Fibrates, the ligands of peroxisome proliferator-activated receptor alpha (PPARalpha), are used as a class of lipid-lowering drugs in clinical practice for the treatment of dyslipidemia. Fibrates are well tolerated in most cases concomitantly with occasional adverse reactions including muscular toxicity, which is enhanced by the combination with statins. This study was designed to investigate the effects of bezafibrate as a PPARalpha agonist on human embryo rhabdomyosarcoma (RD) cells and possible mechanisms responsible for bezafibrate mediated myopathy. The results revealed that bezafibrate caused a dose-dependent decrease in cell viability, which was fortified in association with atorvastatin at a pharmacological dose. Bezafibrate at toxic doses of 300 and 1000microM upregulated PPARalpha at the mRNA level, counteracted by a PPARalpha antagonist (MK886). Bezafibrate at a toxic dose induced typical apoptotic characteristics related to the inhibition of phosphorylation of Akt which was blocked by PPARalpha antagonist. Toxic doses of bezafibrate initiated a significant increase in pyruvate dehydrogenase kinase 4 mRNA and protein levels, compromised by MK886. These results suggest the critical roles of PPARalpha signaling in bezafibrate induced myotoxicity and the involvement of apoptosis through Akt pathway. PMID- 19683051 TI - A second generation multiplex PCR for typing strains of Neospora caninum using six DNA targets. AB - Genetic diversity of Neospora caninum was investigated through a study of repetitive sequences found in the genome of this species. Twenty different loci were studied, and three were identified that varied in repeat content amongst isolates. No relationship was found between the copy number of repetitive sequences present and host type or geographical location from which the isolates were derived. A multiplex PCR assay was developed for multilocus-strain typing using three microsatellites and three minisatellites, based on the polymorphisms found in the repetitive sequences. This study therefore extends knowledge on the repetitive sequences found in the N. caninum genome and the diversity found within the species. It also provides a second generation multiplex assay that can be used to study the biology of N. caninum. In addition, this study included Neospora hughesi (along with other closely related apicomplexans) as controls. The present study shows N. hughesi to be quite distinct from N. caninum in these repetitive sequences, thereby potentially providing a new approach for the differentiation of these two taxa. PMID- 19683052 TI - How the head rules the heart: the chemical neuroanatomy of the pathways of cardiovascular regulation. PMID- 19683053 TI - Thermotolerance and gene expression following heat stress in the whitefly Bemisia tabaci B and Q biotypes. AB - The whitefly Bemisia tabaci (Gennadius) causes tremendous losses to agriculture by direct feeding on plants and by vectoring several families of plant viruses. The B. tabaci species complex comprises over 10 genetic groups (biotypes) that are well defined by DNA markers and biological characteristics. B and Q are amongst the most dominant and damaging biotypes, differing considerably in fecundity, host range, insecticide resistance, virus vectoriality, and the symbiotic bacteria they harbor. We used a spotted B. tabaci cDNA microarray to compare the expression patterns of 6000 ESTs of B and Q biotypes under standard 25 degrees C regime and heat stress at 40 degrees C. Overall, the number of genes affected by increasing temperature in the two biotypes was similar. Gene expression under 25 degrees C normal rearing temperature showed clear differences between the two biotypes: B exhibited higher expression of mitochondrial genes, and lower cytoskeleton, heat-shock and stress-related genes, compared to Q. Exposing B biotype whiteflies to heat stress was accompanied by rapid alteration of gene expression. For the first time, the results here present differences in gene expression between very closely related and sympatric B. tabaci biotypes, and suggest that these clear-cut differences are due to better adaptation of one biotype over another and might eventually lead to changes in the local and global distribution of both biotypes. PMID- 19683055 TI - Atorvastatin reverses cardiac remodeling possibly through regulation of protein kinase D/myocyte enhancer factor 2D activation in spontaneously hypertensive rats. AB - The present study was designed to determine whether atorvastatin reduced hypertension-induced cardiac remodeling and whether these effects involved Protein Kinase D (PKD) and Myocyte Enhancer Factor 2D (MEF2D), factors known to be implicated in cardiac hypertrophy and fibrosis. 16-Week-old spontaneously hypertensive rats (SHRs) and age-matched Wistar-Kyoto (WKY) rats were included. Blood pressure and serum lipid concentration were measured. H-E staining, myocardial transverse diameter, and echocardiography were examined to evaluate cardiac hypertrophy. Hydroxyproline content assay and Masson's trichrome staining were used to estimate cardiac fibrosis. Atorvastatin (10, 25 and 50mg/kg/day) was administered for 8 weeks. Increased blood pressure and cardiac remodeling were prominent in SHRs compared with WKY rats. SHRs also had elevated PKD and MEF2D activation. The systolic blood pressure, myocardial transverse diameter and hydroxyproline content were positively correlated with the activation level of PKD and MEF2D in SHRs. Atorvastatin significantly attenuated the activation of PKD and MEF2D. It may be concluded that atorvastatin reverses hypertension induced cardiac remodeling partially through down-regulation of PKD/MEF2D activation. Our results predict novel therapeutic targets for atorvastatin in treating hypertensive patients. PMID- 19683054 TI - beta2-Agonist modulates epithelial gene expression involved in the T- and B-cell chemotaxis and induces airway sensitization in human isolated bronchi. AB - Regular use of beta(2)-adrenoceptor agonists may enhance non-specific airway responsiveness and inflammation. In earlier experimental studies, we showed that prolonged in vitro fenoterol exposure induced airway sensitization via perturbed epithelial regulation of bronchoconstriction. The aim of the present work was to examine the involvement of inflammatory mediator genes and proinflammatory cells and to investigate the role of the bronchial epithelium in these untoward effects. Bronchial tissues were surgically removed from 17 ex-smokers. Bronchial rings and primary cultures of bronchial epithelial cells were incubated with 0.1microM fenoterol for 15h. Levels of mRNA-expression were analyzed using a real time quantitative reverse transcription-polymerase chain reaction array. Bronchial rings were contracted with endothelin-1 and immune cell infiltration was assessed by immunohistochemistry. Compared to paired controls, fenoterol up regulated the mRNAs of cytokines/proteins implicated in the recruitment of T and B cells or the activation and proliferation of bronchial epithelial cells (CCL20/MIP-3alpha, FOXA2, PPAR-gamma) in isolated bronchi and in cultured epithelial cells. Fenoterol exposure significantly enhanced CD8(+)-T and differentiated CD138(+)-B-cells infiltration into the bronchi, especially the subepithelial area. Increase in CD8 or CD138 labeling-intensity strongly correlated with rise in maximal contraction to endothelin-1 induced by fenoterol exposure. In summary, our results show that fenoterol modulates the T and B cells chemotaxis possibly via the epithelial chemokine secretion in isolated bronchi from ex-smokers. They also suggest that the infiltration of resident T and B cells into the subepithelial area is associated with an increase in airway responsiveness due to fenoterol exposure. PMID- 19683056 TI - Functional magnetic resonance imaging of awake behaving macaques. AB - In recent years, more and more laboratories have developed functional Magnetic Resonance Imaging (fMRI) for awake non-human primates. This research is essential to provide a link between non-invasive hemodynamic signals recorded in the human brain and the vast body of knowledge gained from invasive electrophysiological studies in monkeys. Given that their brain structure is so closely related to that of humans and that monkeys can be trained to perform complicated behavioral tasks, results obtained with monkey fMRI and electrophysiology can be compared to fMRI results obtained in humans, and provide information crucial to a better understanding of the mechanisms by which different cortical areas perform their functions in the human brain. However, despite that the first publications on fMRI in awake behaving macaques appeared approximately 10 years ago (Logothetis et al. (1999) [1], Stefanacci et al. (1998) [2], Dubowitz et al. (1998) [3]), relatively few laboratories perform such experiments routinely, a sign of the significant technical difficulties that must be overcome. The higher spatial resolution required because of the animal's smaller brain results in poorer signal-to-noise ratios than in human fMRI, which is further compounded by problems due to animal motion. Here, we discuss the specific challenges and benefits of fMRI in the awake monkey and review the methodologies and strategies for scanning behaving macaques. PMID- 19683057 TI - One-step expression and purification of single-chain variable antibody fragment using an improved hexahistidine tag phagemid vector. AB - Millions of candidate clones are commonly obtained following rounds of phage displayed antibody library panning, and expression of those selected single-chain variable fragment (scFv) is required for secondary functional screening to identify positive clones. Large scale functional screening is often hampered by the time-consuming and labor-intensive subcloning of those candidate scFv clones into a bacterial expression vector carrying an affinity tag for scFv purification and detection. To overcome the limitations and to develop a multiplex approach, an improved hexahistidine tag phagemid vector was constructed for one-step scFv expression and purification. By using hexahistidine as an affinity tag, soluble scFvs can be rapidly and cost-effectively captured from Escherichia coli periplasmic extracts. For proof-of-concept, feasibility of the improved phagemid vector was examined against two scFvs, L17E4d targeting a cell surface antigen and L18Hh5 recognizing a monoclonal antibody (mAb). Using 1 ml of Ni-NTA agarose, 0.2-0.5 mg of soluble scFv was obtained from 1 L of bacteria culture, and the purified scFvs bound specifically to their target antigens with high affinity. Moreover, using two randomly selected hapten-specific scFv phage clones, it was demonstrated that the display of scFvs on phage surface was not affected by the hexahistidine affinity tag. These results suggest the improved phagemid vector allows the shuttle of phage-displayed antibody library panning and functional scFv production. Importantly, the improved phagemid vector can be easily adapted for multiplex screening. PMID- 19683058 TI - Immune response and gene expression in shrimp (Litopenaeus vannamei) hemocytes and hepatopancreas against some pathogen-associated molecular patterns. AB - The effects of some pathogen-associated molecular patterns (PAMPs) (laminarin, LPS and poly I:C) on total hemocyte counts (THC), phenoloxidase (PO) activity, superoxide anion production and lectin, prophenoloxidase, lysozyme, cytosolic manganese superoxide dismutase (C-MnSOD) and catalase (CAT) gene expression were studied. The results showed that the production or activity of most tested immune factors and the expression of most tested genes were up-regulated after stimulation with PAMPs, among which the highest value of lectin with 4.4 times as much as that of the control group appeared at 6 h in hemocytes, of CAT with 47 times as much as that of the control group appeared at 12 h in hepatopancreas, and with 2.7 times higher than that of the control group at 24 h of C-MnSOD in hepatopancreas after laminarin injection. The peak value of proPO, lysozyme and C MnSOD appeared at 6 h in hepatopancreas, 24 h in hepatopancreas and 24 h in hemocytes after LPS injection, respectively. The highest expression level of lysozyme appeared at 12 h in hemocytes after poly I:C injection. However, significant decreases of PO activity in hemocytes and lectin expression in hepatopancreas were found after poly I:C injection, and a dramatic down regulation of proPO expression from 3 h to 48 h was found in hemocytes after injection with laminarin, LPS and poly I:C. The results suggest that the shrimp immune response could be activated or inhibited by different PAMPs, and that the hepatopancreas also plays a key role by synthesizing immune factors. PMID- 19683059 TI - Met carriers of BDNF Val66Met genotype show increased N-acetylaspartate concentration in the anterior cingulate cortex. AB - Decreased levels of N-acetylaspartate (NAA) and brain-derived neurotrophic factor (BDNF) in the anterior cingulate cortex (ACC) have been linked to neuronal loss and psychiatric disorders like schizophrenia and bipolar disorder. We previously found that BDNF serum concentration was predicted by the concentration of NAA in the ACC, indicating that neuronal integrity and vitality of a cortical region like the ACC, as reflected by a high concentration of NAA, might be related to high concentrations of BDNF in serum. Moreover, our recent finding that Val66Met genotype appears to predict the BDNF serum level in healthy human volunteers suggests the Met allele to be connected to higher concentrations of BDNF in serum. We examined absolute NAA concentrations in the ACC and hippocampus of 40 male and 42 female healthy volunteers (age: 33.3+/-9 years). We found NAA in the ACC to be significantly increased in Met carriers (F=5.2, df=1, p=0.025). On the other hand, the concentration of creatine+phosphocreatine in the hippocampus was significantly decreased in Met carriers. We hypothesize that higher NAA levels in the ACC might contribute to the protection of Met allele carriers against major psychiatric disorders as schizophrenia and bipolar disorder. PMID- 19683061 TI - A hybrid approach to automatic clustering of white matter fibers. AB - Recently, the tract-based white matter (WM) fiber analysis has been recognized as an effective framework to study the diffusion tensor imaging (DTI) data of human brain. This framework can provide biologically meaningful results and facilitate the tract-based comparison across subjects. However, due to the lack of quantitative definition of WM bundle boundaries, the complexity of brain architecture and the variability of WM shapes, clustering WM fibers into anatomically meaningful bundles is nontrivial. In this paper, we propose a hybrid top-down and bottom-up approach for automatic clustering and labeling of WM fibers, which utilizes both brain parcellation results and similarities between WM fibers. Our experimental results show reasonably good performance of this approach in clustering WM fibers into anatomically meaningful bundles. PMID- 19683060 TI - Dynamic changes in white and gray matter volume are associated with outcome of surgical treatment in temporal lobe epilepsy. AB - BACKGROUND: The reasons for surgical failure in 30% of patients with unilateral mesial temporal lobe epilepsy (MTLE) are still unclear. We investigated if different outcomes could be associated to different patterns of subtle gray matter atrophy (GMA) and white matter atrophy (WMA), and searched for postoperative magnetic resonance imaging (MRI) changes. METHODS: We studied 69 controls and 67 operated patients with refractory unilateral MTLE. Patients were grouped as seizure-free (SF) group (34 patients Engel's IA), worthwhile improvement group (23 patients, Engel's IB-IIA) and failure group (10 patients Engel's IIB-IV). We created a voxel-based morphometry/MATLAB code to mask the surgical lacuna, and performed t-test and paired t-test to evaluate preoperative and postoperative MRI scans. RESULTS: Failure group showed a widespread pattern of preoperative GMA. On SF and improvement groups we identified a more restricted pattern of GMA. The three groups presented a widespread, bilateral pattern of WMA. In contrast, postoperative analyses showed bilateral hemispheric recovery (a relative increase of WM concentration) on SF and improvement groups, but few changes on failure group. We also identified areas with relative postoperative increase of GM on both SF and improvement groups, more widespread on SF group. CONCLUSION: Areas of subtle GMA may be related to poorer surgical outcome. In addition, we demonstrated a postoperative relative increase of WM and GM concentration associated with seizure control. These changes may represent neuroplasticity related to improvement of brain function after seizure control. Further studies with a multimodal approach may help to predict surgical outcome and improve selection of patients for surgical treatment of MTLE. PMID- 19683062 TI - Dysfunctional brain circuitry in obsessive-compulsive disorder: source and coherence analysis of EEG rhythms. AB - BACKGROUND: Morphological and functional studies suggested involvement of several cortical and subcortical circuitries in patients with obsessive-compulsive disorder (OCD). The aim of the present study was to investigate networks involved in OCD pathophysiology, using power (coupling of EEG bands, low-resolution electromagnetic tomography-LORETA) and coherence analysis in drug-naive patients. METHOD: EEG was obtained from 37 drug-naive patients with OCD and 37 age- and sex matched controls. Resting EEG was recorded from 29 scalp channels. Coupling (ratio and correlation) between low and high frequencies was analyzed on Fz. For each frequency band, LORETA current density distribution, intra-hemispheric and inter-hemispheric coherence analysis were computed. RESULTS: OCD had increased current density for delta in the insula and for beta in frontal, parietal and limbic lobes. OCD also had decreased inter-hemispheric coherence and reduced coupling between delta and beta frequencies. CONCLUSIONS: In OCD, increased frontal beta is consistent with previous evidence of frontal dysfunction. Hyperactivity of insular delta sources, together with rhythms decoupling and reduced interhemispheric alpha coherence are consistent with additional involvement of cortico-subcortical functional connections. Combined use of power and coherence analysis may provide functional measures on different levels of involvement of cortico-subcortical circuits in neuropsychiatric disorders. PMID- 19683063 TI - The human parahippocampal cortex subserves egocentric spatial learning during navigation in a virtual maze. AB - BACKGROUND: Present evidence suggests that the hippocampus (HC) and the parahippocampal cortex (PHC) are involved in allocentric (world-centered) spatial memory. However, the putative role of the PHC in egocentric (body-centered) spatial learning has received only limited systematic investigation. METHODS: To examine the role of the PHC in egocentric learning, 19 healthy volunteers learned to find their way in a virtual maze during functional magnetic resonance imaging (fMRI). The virtual maze presented a first-person view, lacked any topographical landmarks and could be learned only using egocentric navigation strategies. RESULTS: During learning, increased medial temporal lobe activity was observed in the PHC bilaterally. Activity was also observed in cortical areas known to project to the PHC and proposed to contribute to egocentric spatial navigation and memory. CONCLUSIONS: Our results point to a role of the PHC for the representation and storage of egocentric information. It seems possible that the PHC contributes to egocentric memory by its feedback projections to the posterior parietal cortex. Moreover, access to allocentric and egocentric streams of spatial information may enable the PHC to construct a global and comprehensive representation of spatial environments and to promote the construction of stable cognitive maps by translating between egocentric and allocentric frames of memory. PMID- 19683064 TI - C. difficile infection (CDI) in a long-term acute care facility (LTAC). AB - BACKGROUND: C. difficile infection (CDI) is a common nosocomial infection in hospitals and impacts increased hospital cost and length of stay. Since scant information is available about the incidence and prevalence of CDI in Long-term acute care hospitals (LTACs), we therefore studied this at one local facility. METHODS: Demographic and other data, and a fresh stool sample were obtained from all new LTAC admissions not carrying a prior diagnosis of CDI during the study period (July 23 to August 22, 2007). A GDH test for C. difficile antigen was performed. All initially positive stools were tested for toxins A and B and a sample was frozen for culture and typing. All antigen-negative patients were monitored for the development of diarrhea during the course of their LTAC hospitalization and, if clinically indicated, a sample was sent for toxins A and B testing and if positive, a stool sample was frozen and stored for culture and typing. Therapy of CDI was noted. RESULTS: 36 patients were admitted during the study period. 4 of 31 (12.9%) of patients tested were antigen (+) on admission of which 2 (6.5%) were asymptomatic carriers and 2 (6.5%) had unsuspected active disease, including one with the BI epidemic strain. In follow-up, 20/36 (55.5%) developed diarrhea of which an additional 5 (13.8%) patients had developed CDI (average, hospital day 38) in the hospital. Therapy was instituted with vancomycin in 5/7 patients and metronidazole in 2/7 patients. During that quarter, the rate of nosocomial acquired CDI was 3.12 per 1000 patient days. CONCLUSIONS: C. difficile carriage and unsuspected clinical CDI occurs, including with the BI epidemic strain disease, in an important minority of patients, which may act as a reservoir for spread. New strategies for detection and prevention of CDI are needed. PMID- 19683065 TI - Differential effect of appendectomy and tonsillectomy on anti-Kudoa sp. antibodies in patients with MALTectomy. AB - We found an association between tonsillectomized patients and subsequent appendicitis. We also observed that MALTectomy significantly decreased secretory IgA levels in serum of patients, being this decrease more pronounced when both operations (tonsillectomy and appendectomy) had been performed. The elevated humoral responses detected previously by us in BALB/c mice immunized with Kudoa sp. pseudocyst extracts and the high IgG1 and IgE levels induced by the oral administration of Kudoa sp. pseudocysts to BALB/c mice showed the possible immunopathological effects in man from the ingestion of Kudoa sp. infected fish. We use the ELISA method to investigate the possible relationship between MALTectomy (tonsillectomy and appendectomy) and specific antibody levels to Kudoa sp. Both anti-Kudoa sp. specific antibody levels and the number of patients that recognized Kudoa sp. antigens were greater in tonsillectomy patients when compared to the control and the other studied groups (appendectomized and appendectomized+tonsillectomies patients). Tonsillectomy was associated to a switch in the class of immunoglobulins involved in these responses and these responses may be abrogated by appendectomy. Tonsils and appendix may respond in different ways to Kudoa sp. antigens and these different reactions may be involved in some immunopathological reactions. PMID- 19683066 TI - What can natural language processing do for clinical decision support? AB - Computerized clinical decision support (CDS) aims to aid decision making of health care providers and the public by providing easily accessible health related information at the point and time it is needed. natural language processing (NLP) is instrumental in using free-text information to drive CDS, representing clinical knowledge and CDS interventions in standardized formats, and leveraging clinical narrative. The early innovative NLP research of clinical narrative was followed by a period of stable research conducted at the major clinical centers and a shift of mainstream interest to biomedical NLP. This review primarily focuses on the recently renewed interest in development of fundamental NLP methods and advances in the NLP systems for CDS. The current solutions to challenges posed by distinct sublanguages, intended user groups, and support goals are discussed. PMID- 19683068 TI - Towards responsible system development in health services: a discourse analysis study of design conflict resolution tactics. AB - We set out to examine design conflict resolution tactics used in development of large information systems for health services and to outline the design consequences for these tactics. Discourse analysis methods were applied to data collected from meetings conducted during the development of a web-based system in a public health context. We found that low risk tactics were characterized by design issues being managed within the formal mandate and competences of the design group. In comparison, high risk tactics were associated with irresponsible compromises, i.e. decisions being passed on to others or to later phases of the design process. The consequence of this collective disregard of issues such as responsibility and legitimacy is that the system design will be impossible to implement in factual health service contexts. The results imply that downstream responsibility issues have to be continuously dealt with in system development in health services. PMID- 19683067 TI - What is biomedical informatics? AB - Biomedical informatics lacks a clear and theoretically-grounded definition. Many proposed definitions focus on data, information, and knowledge, but do not provide an adequate definition of these terms. Leveraging insights from the philosophy of information, we define informatics as the science of information, where information is data plus meaning. Biomedical informatics is the science of information as applied to or studied in the context of biomedicine. Defining the object of study of informatics as data plus meaning clearly distinguishes the field from related fields, such as computer science, statistics and biomedicine, which have different objects of study. The emphasis on data plus meaning also suggests that biomedical informatics problems tend to be difficult when they deal with concepts that are hard to capture using formal, computational definitions. In other words, problems where meaning must be considered are more difficult than problems where manipulating data without regard for meaning is sufficient. Furthermore, the definition implies that informatics research, teaching, and service should focus on biomedical information as data plus meaning rather than only computer applications in biomedicine. PMID- 19683069 TI - Comparing early outbreak detection algorithms based on their optimized parameter values. AB - BACKGROUND: Many researchers have evaluated the performance of outbreak detection algorithms with recommended parameter values. However, the influence of parameter values on algorithm performance is often ignored. METHODS: Based on reported case counts of bacillary dysentery from 2005 to 2007 in Beijing, semi-synthetic datasets containing outbreak signals were simulated to evaluate the performance of five outbreak detection algorithms. Parameters' values were optimized prior to the evaluation. RESULTS: Differences in performances were observed as parameter values changed. Of the five algorithms, space-time permutation scan statistics had a specificity of 99.9% and a detection time of less than half a day. The exponential weighted moving average exhibited the shortest detection time of 0.1 day, while the modified C1, C2 and C3 exhibited a detection time of close to one day. CONCLUSION: The performance of these algorithms has a correlation to their parameter values, which may affect the performance evaluation. PMID- 19683070 TI - Extending the Fellegi-Sunter probabilistic record linkage method for approximate field comparators. AB - Probabilistic record linkage is a method commonly used to determine whether demographic records refer to the same person. The Fellegi-Sunter method is a probabilistic approach that uses field weights based on log likelihood ratios to determine record similarity. This paper introduces an extension of the Fellegi Sunter method that incorporates approximate field comparators in the calculation of field weights. The data warehouse of a large academic medical center was used as a case study. The approximate comparator extension was compared with the Fellegi-Sunter method in its ability to find duplicate records previously identified in the data warehouse using different demographic fields and matching cutoffs. The approximate comparator extension misclassified 25% fewer pairs and had a larger Welch's T statistic than the Fellegi-Sunter method for all field sets and matching cutoffs. The accuracy gain provided by the approximate comparator extension grew as less information was provided and as the matching cutoff increased. Given the ubiquity of linkage in both clinical and research settings, the incremental improvement of the extension has the potential to make a considerable impact. PMID- 19683071 TI - Direct pancreatoscopy with an ultrathin forward-viewing endoscope in intraductal papillary mucinous neoplasm of the pancreas. PMID- 19683072 TI - Hepatitis B virus genotype C is associated with more severe liver fibrosis than genotype B. AB - BACKGROUND & AIMS: Histologic analyses of liver fibrosis have been limited by small sample sizes and the predominance of samples from patients with active hepatitis. METHODS: We performed a prospective study of transient elastography in treatment-naive patients with chronic hepatitis B, to investigate the relationship between hepatitis B virus (HBV) genotype and liver fibrosis. A validated liver stiffness measurement algorithm was used to define insignificant fibrosis and advanced fibrosis. RESULTS: Of 1106 patients, 711 (64%) were older than age 40, 370 (34%) had positive test results for hepatitis B e antigen (HBeAg), and 386 (35%) had increased serum levels of alanine aminotransferase. Of the patients, 524 (49%) had genotype B and 582 (51%) had genotype C HBV infection. Patients with genotype C infection had insignificant fibrosis less often (42% vs 55%; P < .0001) and advanced fibrosis more often (25% vs 19%; P = .015) than those infected with genotype B HBV. The difference in the severity of liver fibrosis between the 2 HBV genotypes was most marked among patients older than age 40 and those who tested negative for HBeAg. The mean age of patients infected by genotype C was greater than that of patients infected by genotype B HBV (41 vs 36 y). Among patients who were older than age 40 and tested negative for HBeAg, those with genotype C infection had higher levels of HBV DNA and alanine aminotransferase than those with genotype B HBV. CONCLUSIONS: Genotype C HBV was associated with more severe liver fibrosis than genotype B HBV, probably because of delayed HBeAg seroconversion and prolonged active disease. PMID- 19683073 TI - Surgery for gastroesophageal reflux disease: esophageal impedance to progress? PMID- 19683074 TI - Shh and Pax6 have unconventional expression patterns in embryonic morphogenesis in Sepia officinalis (Cephalopoda). AB - Cephalopods show a very complex nervous system, particularly derived when compared to other molluscs. In vertebrates, the setting up of the nervous system depends on genes such as Shh and Pax6. In this paper we assess Shh and Pax6 expression patterns during Sepia officinalis development by whole-mount in situ hybridization. In vertebrates, Shh has been shown to indirectly inhibit Pax6. This seems to be the case in cephalopods as the expression patterns of these genes do not overlap during S. officinalis development. Pax6 is expressed in the optic region and brain and Shh in gut structures, as already seen in vertebrates and Drosophila. Thus, both genes show expression in analogous structures in vertebrates. Surprisingly, they also exhibit unconventional expressions such as in gills for Pax6 and ganglia borders for Shh. They are also expressed in many cephalopods' derived characters among molluscs as in arm suckers for Pax6 and beak producing tissues, nuchal organ and neural cord of the arms for Shh. This new data supports the fact that molecular control patterns have evolved with the appearance of morphological novelties in cephalopods as shown in this new model, S. officinalis. PMID- 19683075 TI - Impact of ageing on muscle cell regeneration. AB - Skeletal muscle regeneration is a coordinate process in which several factors are sequentially activated to maintain and preserve muscle structure and function. The major role in the growth, remodeling and regeneration is played by satellite cells, a quiescent population of myogenic cells that reside between the basal lamina and plasmalemma and are rapidly activated in response to appropriate stimuli. However, in several muscle conditions, including aging, the capacity of skeletal muscle to sustain an efficient regenerative pathway is severely compromised. Nevertheless, if skeletal muscle possesses a stem cell compartment it is not clear why the muscle fails to regenerate under pathological conditions. Either the resident muscle stem cells are too rare or intrinsically incapable of repairing major damage, or perhaps the injured/pathological muscle is a prohibitive environment for stem cell activation and function. Although we lack definitive answers, recent experimental evidences suggest that the mere presence of endogenous stem cells may not be sufficient to guarantee muscle regeneration, and that the presence of appropriate stimuli and factors are necessary to provide a permissive environment that permits stem cell mediated muscle regeneration and repair. In this review we discuss the molecular basis of muscle regeneration and how aging impacts stem cell mediated muscle regeneration and repair. PMID- 19683076 TI - The myelin-associated oligodendrocytic basic protein (MOBP) as a relevant primary target autoantigen in multiple sclerosis. AB - Multiple sclerosis (MS) is a disease of the human CNS, characterized by perivascular inflammation, demyelination and axonal damage. Although the etiology of MS is unknown, it is believed that the disease results from destructive autoimmune mechanisms, presumably initiated by abnormal activation of potentially pathogenic autoimmune T-cells recognizing CNS components. The myelin-associated oligodendrocyte basic protein (MOBP), a relatively abundant CNS-specific myelin protein, which plays a role in stabilizing the myelin sheath in the CNS, has recently been implicated in the pathogenesis of MS. Here we review studies showing that MOBP is as an important candidate target antigen in MS as the other widely studied target antigens, myelin basic protein (MBP), proteolipid protein (PLP), and myelin oligodendrocyte glycoprotein (MOG). The studies summarized below indicate that T-cell autoimmunity against MOBP can be detected in MS patients; T-cells reactive against MOBP can be pathogenic in several mouse strains as well as in the "humanized" HLA-DR15-Tg mice; and, that the HLA-DQ6 restricted, but not HLA-DR15-restricted, MOBP-reactive T-cells cause in HLA-DR15 Tg mice MS-like clinical disease associated with perivascular and parenchymal infiltration, demyelination, axonal loss, and optic neuritis. Accordingly, the MOBP should be considered a bona fide primary target antigen in MS, in addition to MBP, PLP, and MOG. PMID- 19683077 TI - The Wnt signaling pathway and rheumatoid arthritis. AB - The Wnt signaling pathways play a key role in cell renewal, and there are two such pathways. In patients with rheumatoid arthritis (RA), the synovial membrane expresses genes such as Wnt and Fz at higher levels than those observed in patients without RA. The Wnt proteins are glycoproteins that bind to receptors of the Fz family on the cell surface. The Wnt/Fz complex controls tissue formation during embryogenesis, as well as throughout the process of limb development and joint formation. Recent studies have suggested that this signaling pathway plays a role in the pathophysiology of RA. Greater knowledge of the role of the Wnt signaling pathway in RA could improve understanding of the differences in RA clinical presentation and prognosis. Further studies should also focus on Wnt family members as molecular targets in the treatment of RA. PMID- 19683079 TI - Kinetics and thermodynamics of the interaction of 1-anilino-naphthalene-8 sulfonate with proteins. AB - Although 1-anilino-naphthalene-8-sulfonate (ANS) has been widely used in protein folding and binding studies, the detailed mechanism of this interaction is not fully understood. In this work the binding of ANS was analyzed at pre-equilibrium and equilibrium conditions using bovine serum albumin (BSA) as model. We employed a combined approach including the analysis of fluorescence, near-UV circular dichroism and isothermal titration calorimetric data. Experiments at equilibrium with these techniques identify three ANS molecules bound at hydrophobic cavities in BSA. Pre-equilibrium fluorescence analysis unambiguously indicated that the binding of ANS at hydrophobic cavities of BSA occurs at two different and independent classes of sites with similar affinities and quantum yields, two features that are undetectable by the equilibrium analysis. The binding of ANS to the first site is thermodynamically favored by similar contributions of the enthalpic (DeltaH = -22 kJ/mol) and entropic terms (-TDeltaS = -17 kJ/mol), while the binding to the second site is enthalpically driven (DeltaH = -31 kJ/mol; TDeltaS = -0.6 kJ/mol). Complementary information from molecular docking showed three ANS molecules bound at hydrophobic cavities in BSA subdomains IIA and IIIA with binding affinities in the order of those found experimentally and three additional ANS molecules bound at water exposed sites. PMID- 19683080 TI - Counting on big numbers. PMID- 19683078 TI - Kinetic characterization of human phosphopantothenoylcysteine synthetase. AB - Phosphopantothenoylcysteine synthetase (PPCS) catalyzes the formation of phosphopantothenoylcysteine from (R)-phosphopantothenate and L-cysteine with the concomitant consumption of a nucleotide triphosphate. Herein, the human coaB gene encoding PPCS is cloned into pET23a and overexpressed in E. coli BL21(DE3), to yield 10mg of purified enzyme per liter of culture. Detailed kinetic studies found that this PPCS follows a similar Bi Uni Uni Bi Ping Pong mechanism as previously described for the E. faecalis PPCS, except that the human enzyme can use both ATP and CTP with similar affinity. One significant difference for human PPCS catalysis with respect to ATP and CTP is that the enzyme shows cooperative binding of ATP, measured as a Hill constant of 1.7. PPCS catalysis under CTP conditions displayed Michaelis constants of 265 microM, 57 microM, and 16 microM for CTP, PPA, and cysteine, respectively, with a kcat of 0.53+/-0.01 s(-1) for the reaction. Taking into account the cooperativity under ATP condition, PPCS exhibited Michaelis constants of 269 microM, 13 microM, and 14 microM for ATP, PPA, and cysteine, respectively, with a kcat of 0.56 s(-1) for the reaction. Oxygen transfer studies found that 18O from [carboxyl-18O] phosphopantothenate is incorporated into the AMP or CMP produced during PPCS catalysis, consistent with the formation of a phosphopantothenoyl cytidylate or phosphopantothenoyl adenylate intermediate, supporting similar catalytic mechanisms under both CTP and ATP conditions. Inhibition studies with GTP and UTP as well as product inhibition studies with CMP and AMP suggest that human PPCS lacks strong nucleotide selectivity. PMID- 19683081 TI - Proteomics meets blood banking: identification of protein targets for the improvement of platelet quality. AB - Proteomics has brought new perspectives to the fields of hematology and transfusion medicine in the last decade. The steady improvement of proteomic technology is propelling novel discoveries of molecular mechanisms by studying protein expression, post-translational modifications and protein interactions. This review article focuses on the application of proteomics to the identification of molecular mechanisms leading to the deterioration of blood platelets during storage - a critical aspect in the provision of platelet transfusion products. Several proteomic approaches have been employed to analyse changes in the platelet protein profile during storage and the obtained data now need to be translated into platelet biochemistry in order to connect the results to platelet function. Targeted biochemical applications then allow the identification of points for intervention in signal transduction pathways. Once validated and placed in a transfusion context, these data will provide further understanding of the underlying molecular mechanisms leading to platelet storage lesion. Future aspects of proteomics in blood banking will aim to make use of protein markers identified for platelet storage lesion development to monitor proteome changes when alterations such as the use of additive solutions or pathogen reduction strategies are put in place in order to improve platelet quality for patients. PMID- 19683082 TI - Alterations in cellular proteome and secretome upon differentiation from monocyte to macrophage by treatment with phorbol myristate acetate: insights into biological processes. AB - Monocyte and macrophage are mainly involved in immune response and inflammatory processes. Monocytes circulate in the bloodstream and migrate to various tissues where they can differentiate to macrophages. However, the molecular basis of biological processes involved in this cellular differentiation remains ambiguous. This study was to investigate alterations in cellular and secreted proteins after this differentiation phase. Macrophage was differentiated from U937 human monocytic cell line by treatment with 100 ng/ml phorbol myristate acetate (PMA) for 48 h. Cellular and secreted proteins extracted from PMA-treated cells (macrophages) were compared with those of untreated cells (monocytes) using 2-DE (n=5 gels/condition; stained with Deep Purple fluorescence dye). Quantitative intensity analysis revealed 81 and 67 protein spots whose levels were significantly altered in cellular proteome and secretome. These proteins were subsequently identified by Q-TOF MS and/or MS/MS analyses. The altered levels of cellular elongation factor-2 (EF-2) and secreted alpha-tubulin were confirmed by Western blot analysis. Global protein network analysis demonstrated that these altered proteins were involved in cell death, lipid metabolism, cell morphology, cellular movement, and protein folding. Our data may provide some insights into molecular mechanisms of biological processes upon differentiation from monocytes to macrophages. PMID- 19683083 TI - Hepcidin assay in serum by SELDI-TOF-MS and other approaches. AB - Hepcidin, a liver peptide hormone, is the central regulator of iron homeostasis. Hepcidin synthesis is modulated by iron stores, so that iron repletion increases its levels to prevent pathological overload, while iron deficiency strongly inhibits hepcidin to allow an increase in iron absorption from duodenal cells. The emerging pivotal role of hepcidin in iron homeostasis, along with its important links with basic pathways like inflammation, makes the availability of an accurate hepcidin assay as a potentially powerful investigative tool to improve our understanding as well as our diagnostic/prognostic capabilities in many human diseases. There has been a great interest worldwide in developing a reliable and widely applicable assay of the hormone in biological fluids. Being optimal for low-molecular-weight biomarkers, SELDI-TOF-MS has emerged as a valid tool for hepcidin assay. Here we review recent results obtained with this technique, as well as with other Mass Spectrometry-based and immunological methods. PMID- 19683084 TI - Chromosomal tethering and proviral integration. AB - Since integration into the host cell genome is an obligatory step for their replication, retro-elements are both potent insertional mutagens and also suitable vectors for gene therapy. Many recent studies reported that the integration process is not random but, on the contrary, higly regulated at the molecular level. Many viral proteins and cellular factors play a key role in the integration step, explaining the reason why different retro-elements display distinct integration profiles. This review describes the recent highlights about integration of retro-elements with particular focus on the mechanisms underlying the specificity of integration target-site selection and the step of chromosomal tethering which preceeds insertion of the provirus. PMID- 19683085 TI - Sonic hedgehog alleviates the inhibitory effects of high glucose on the osteoblastic differentiation of bone marrow stromal cells. AB - To assess the influence of high extracellular glucose levels on the osteogenic differentiation of bone marrow stromal cells (BMSCs) and to determine if Sonic hedgehog (Shh) protein can alleviate those effects. BMSCs were incubated with NG (normal glucose), NG+Shh (200 ng/ml Shh in normal glucose), NG+Shh+Gan (200 ng/ml Shh and 5 micromol/L GANT61 in normal glucose), HG (high glucose), HG+Shh (200 ng/ml Shh in high glucose), and HG+Shh+Gan (200 ng/ml Shh and 5 micromol/L GANT61 in high glucose). The expression levels of Shh signaling pathway genes Patched 1 (PTCH1) and osteogenesis-related genes were tested, which included bone morphogenetic protein 4 (BMP4), runt-related transcription factor 2 (Runx2), and osteopontin (OPN). Alkaline phosphatase (ALPase) activity and mineralized matrix formation were also investigated. Immunofluorescent staining of Gli1 was tested for Shh signaling activation. We found that recombinant Shh in normal-glucose medium could promote osteogenic differentiation of BMSCs, while inhibiting Shh signaling by GANT61 could antagonize this differentiation. Besides that high glucose impaired the Shh signaling as well as osteogenic differentiation of BMSCs, reactivation of Shh signal pathway by addition of Shh protein could mitigate the inhibition while further deactivation by Shh inhibitor GANT61 could retain their osteogenic inhibitions. The above data suggest that Shh pathway activity is involved in the HG condition mediated osteoblastic differentiation deficiency for BMSCs and that recombinant Shh could alleviate this inhibitory effect. PMID- 19683086 TI - Antibody kinetics among 8-10 years old respondents to hepatitis B vaccination in a low endemic country and the effect of a booster dose given 5 or 10 years later. AB - Few data are available concerning the persistence of anti-HBs and the effect of booster doses given several years post-vaccination against hepatitis B during preadolescence. The objective of this open-labelled clinical trial was to evaluate the persistence of antibodies after vaccination with three paediatric doses of Engerix-B at the age of 8-10 years and the effect of a booster dose given 5 (Group Y5) or 10 (Group Y10) years later. Anti-HBs were measured before and one month post-primary vaccination, then 5 and 10 years later, before the booster dose, as well as one month and 1 year post-booster. The anamnestic response was defined as a >or=fourfold increase of anti-HBs post-booster (>or=10 IU/L) when compared to pre-booster. Ten years post-primary vaccination, 559 of the 652 initially randomized subjects (86%) were eligible for analysis. Group Y5, 5 years post-booster results: 99% of subjects had detectable levels of antibodies and 96% a titer >or=10 IU/L. The anti-HBs GMTs decreased from 114,489 IU/L one month post-booster to 3354 IU/L 5 years later. Group Y10 results: 10 years post primary vaccination 96% of subjects had a detectable level of anti-HBs and 85% were above the threshold of 10 IU/L. The GMTs one month post-booster were 31,030 IU/L. The challenge with a booster demonstrated an anamnestic response in 99% of subjects in group Y5 and 100% of subjects in group Y10. All subjects were anti HBc negative. The booster doses were well tolerated. The excellent anamnestic response observed after the booster dose demonstrates the persistence of immunity in virtually all young adults vaccinated at the age of 8-10 with three paediatric doses of Engerix-B. PMID- 19683088 TI - Women's health. Preface. PMID- 19683089 TI - A historical perspective of the women's health nurse practitioner. AB - There are more than 12,000 women's health nurse practitioners (WHNPs) currently certified by the National Certification Corporation (NCC) and practicing in a wide range of roles. The purpose of this article is to describe the historical development of the WHNP specialty, and to review the evolution of the specialty from an initially very focused practice in the area of family planning into obstetric and gynecologic care to today's more diffuse role inclusive of primary care. Women's health nurse practitioners must broaden their educational background to include the lifespan of women, not just the reproductive years. With the inclusion of chronic disease management of the middle-aged and elderly woman, WHNPs will provide more comprehensive and integrative health care to women in all areas of the United States. PMID- 19683087 TI - Immunogenicity and tolerability of an AS03(A)-adjuvanted prepandemic influenza vaccine: a phase III study in a large population of Asian adults. AB - The immunogenicity and lot-to-lot consistency of an AS03-adjuvanted H5N1 vaccine were evaluated in 1206 Asian adults, randomised to receive two doses of adjuvanted (3.75 microg haemagglutinin) or diluent-mixed vaccines, 21 days apart. Post-Dose 2, 96.0% of vaccinees in the H5N1-AS03 group demonstrated a four-fold increase in neutralising antibody titres against the vaccine strain A/Vietnam/1194/2004 and 91.4% against strain A/Indonesia/05/2005. Haemagglutination-inhibiting antibodies (titre > or = 1:40) against A/Vietnam/1194/2004 and A/Indonesia/05/2005 strains were observed in 94.3% and 50.2% of subjects, respectively. Lot-to-lot consistency of the AS03-adjuvanted vaccine combinations was demonstrated. The AS03-adjuvanted vaccine was well tolerated, induced a high frequency of immune responses to the vaccine strain, allowed antigen sparing and promoted cross-clade immunity. These characteristics make it suitable for presumptive use if an H5N1 pandemic were considered to be imminent. PMID- 19683090 TI - Promoting wellness in women across the life span. AB - For the purposes of this article, wellness is defined as an individual's subjective experience of overall life satisfaction in relation to physical, mental, emotional, spiritual, social, economic, occupational, and environmental dimensions. Women's wellness focuses on those aspects of well-being that pertain disproportionately, or solely, to women. Wellness includes but is not limited to physical, emotional and social aspects and disruptions that alter a woman's quality of life, such as reproductive and hormonal issues, bone health, gastrointestinal stress, and urinary incontinence. This article discusses women's wellness through the life span, from preconception through death, and considers the implications of these issues for the nursing profession. PMID- 19683091 TI - Human papillomavirus and the HPV vaccine: are the benefits worth the risks? AB - Human papillomavirus (HPV) is the most commonly sexually transmitted infection in the United States. This article gives an overview and discussion of HPV virus types and transmission, and the quadrivalent vaccine now available to protect against it. Included are the nursing implications for the HPV vaccine related to education and counseling of parents, patients, and young adult women regarding HPV vaccination, for whom the vaccine is indicated. PMID- 19683092 TI - The role of nursing in the management of unintended pregnancy. AB - This article explores the role of nurses in the prevention, management, and treatment of unintended pregnancy. All nurses have a responsibility to understand the importance of reproductive health care in the primary care of women and their families, and to be prepared to respond to patients' needs for the prevention and management of unintended pregnancy. A public health framework provides an opportunity to identify the role of the nurse in primary, secondary, and tertiary prevention strategies that can contribute to the management of unintended pregnancy for the health of women and their families. Nursing education and the role of nurses in advocacy for reproductive health concerns are also addressed. PMID- 19683093 TI - Cardiac health: primary prevention of heart disease in women. AB - Heart disease is the number one cause of death among women. Although 450,000 women die annually from heart disease, this fact is unknown to many women. Because heart disease is frequently preventable, increasing awareness of personal risk and preventative measures is a key element of health care for women. Nurse clinicians can evaluate, educate, and counsel women regarding their risk for this pervasive disease and promote behavior changes that will decrease that risk. Research evidence supports that lifestyle behaviors are the cornerstone of heart disease prevention. This article presents current evidence for the prevention of heart disease related to dietary intake, physical activity, weight management, smoking cessation, blood pressure control, and lipid management. Guidelines for implementing findings in clinical practice are discussed. PMID- 19683094 TI - Genetics: breast cancer as an exemplar. AB - Genetic testing for adult-onset diseases is now available. One such test is for the mutations present in the BRCA gene that result in a significantly higher risk for the development of breast cancer or ovarian cancer. Women who have one of these mutations face difficult choices in terms of increased surveillance or prophylactic surgeries. Examining experiences of women with BRCA mutations can serve as an exemplar for other populations at risk for genetically associated adult-onset diseases. PMID- 19683096 TI - Women's mental health: depression and anxiety. AB - Sex ratios for selected mental disorders such as major depressive disorder and anxiety disorder are much higher in women than men. Anxiety disorders constitute the most prevalent mental disorder in adults, and affect twice as many women as men. Depression and anxiety exist comorbidly and along with other mental disorders. This article focuses on depression and anxiety in women, and other conditions comorbid with depression or anxiety: cardiac disease, obesity, vitamin D deficiency, and irritable bowel syndrome. PMID- 19683095 TI - Computer-mediated patient education: opportunities and challenges for supporting women with ovarian cancer. AB - A majority of women with ovarian cancer will face recurrent disease despite receiving aggressive chemotherapy at the time of diagnosis. Given the complex medical and psychosocial needs of women with ovarian cancer and the time constraints within busy clinical settings, providing women with the necessary education related to their disease and treatments can be challenging. The advent of computers and web-based technologies has created new opportunities for educating cancer patients and supporting them to better cope with their disease. This article reviews prior studies of computer-based patient education interventions to identify key intervention components and other factors associated with improved patient outcomes. Opportunities for using computer-based technologies to support women with ovarian cancer are discussed and WRITE Symptoms (a Written Representational Intervention To Ease Symptoms), a web-based, symptom management intervention for women with recurrent ovarian cancer, is introduced. PMID- 19683097 TI - Women prisoners: health issues and nursing implications. AB - The purpose of this article is to describe health issues of women prisoners, analyze the implications of these issues for nursing practice, and consider strategies to improve the health of this vulnerable population. The article focuses primarily on women prisoners in the United States and includes a brief contextual background to explain the rapid increase in their numbers. Although the incarceration of women is increasingly a global problem, the authors focus primarily on the situation in the United States. PMID- 19683099 TI - Epigenetic control of gene expression. PMID- 19683098 TI - Global women's health: a spotlight on caregiving. AB - Caregiving is a women's health issue globally, as many more women than men are informal caregivers. Caregiving related to gender role socialization, burden, and economic and health consequences has been discussed in the literature. Together this body of work demonstrates some positive but mainly negative consequences to the health and economic circumstances of women. Overall achievement of the United Nations' Millennium Development Goals has important implications for informal caregivers globally, because achievement of these goals is essential to reducing the undue burden, the lost opportunities, and the injustice of health care systems that take advantage of women's volunteer caregiving. PMID- 19683100 TI - Hepcidin in acute iron toxicity. AB - BACKGROUND: Hepcidin regulates extracellular iron concentration by inhibiting iron release from macrophages and preventing iron absorption in the intestine. Our objective was to evaluate the expression of hepcidin in the liver in acute iron poisoning in a rat model. METHODS: Male Wistar rats were assigned to group 1, who received 750 mg/kg elemental iron (LD(50)) by gavage, and group 2 (control), who received distilled water. Iron concentrations and liver transaminases were measured in the serum. Hepcidin messenger RNA levels were measured in the liver. RESULTS: Mean serum iron levels, aspartate aminotransferase, alanine aminotransferase, and uric acid were significantly higher in group 1 compared to group 2 (P < .0001, P = .01, P < .0001, and P = 0.0001, respectively). Hepcidin messenger RNA levels in the liver were significantly higher in the study group (P = .005). CONCLUSIONS: In acute iron intoxication, hepcidin expression in the liver significantly increased. Further studies are needed to determine whether hepcidin levels correlate with the severity of the intoxication. PMID- 19683101 TI - Can D-dimer become a new diagnostic parameter for acute appendicitis? AB - INTRODUCTION: In this study, we investigated D-dimer serum level as a diagnostic parameter for acute appendicitis. MATERIALS AND METHODS: Forty-nine patients were enrolled in the study. Patients were classified according to age; sex; duration between the beginning of pain and referral to a hospital or clinic; Alvarado scores; and in physical examination, presence of muscular defense, the number of leukocytes, preoperative ultrasonography, and D-dimer levels of histopathologic study groups were analyzed. RESULTS: Of the patients enrolled in the study, 26.5% were females and 73.5% males. The average age was 21 years (range, 16-38 years) and 81.7% acute appendicitis (AA). According the duration of pain, 63.2% of the patients were referred to the hospital within the first 24 hours, 26.5% of the patients were referred to the hospital within 24 to 48 hours, and 10.3% were referred to the hospital within a period of more than 48 hours. No statistically significant difference was determined regarding D-dimer levels between the histopathologic study groups (P > .05). Alvarado scores lower than 7 were found in 36.7% and 7 or higher in 63.3% of the patients. There was no statistically significant difference related with D-dimer levels between histopathologic study groups (P > .05). The ratio of cases with a number of leukocytes below the upper limit were determined respectively as 32.7% and 67.3%, and no statistically significant difference was found regarding d-dimer levels between histopathologic study groups (P > .05). CONCLUSION: Increased D-dimer levels should not be considered as a diagnostic parameter in diagnosis of acute appendicitis. PMID- 19683103 TI - Emergency physicians report infrequent use of alpha-blockade for the treatment of ureteral stones. AB - PURPOSES: We sought to determine how frequently tamsulosin is used by emergency physicians (EPs) for the treatment of patients with ureteral stones, and examine factors influencing its use. BASIC PROCEDURES: We distributed online and paper surveys to 240 EPs in five states in the United States. Surveys asked the length of time in practice, the type of hospital setting, and the self-reported frequency of tamsulosin use by quartile. FINDINGS: A total of 103 EPs (43%) responded to the survey. The majority (60%, 95% CI 51% to 69%) reported the use of tamsulosin in fewer than 25% of patients presenting with ureteral stones, and many (27%, 95% CI 20% to 36%) had not heard of the use of tamsulosin for urinary stone passage. Practice in a county hospital was associated with infrequent tamsulosin use (p=.02). CONCLUSIONS: Despite evidence that alpha-blockade may be beneficial to patients presenting to the emergency department with ureteral stones, this approach is still used inconsistently by EPs. PMID- 19683102 TI - Thoracic impedance vs chest radiograph to diagnose acute pulmonary edema in the ED. AB - OBJECTIVE: We sought to investigate the relationship between thoracic impedance (Zo) and pulmonary edema on chest radiography in patients presenting to the emergency department (ED) with signs and symptoms of acute decompensated heart failure (ADHF). DESIGN: This was a prospective, blinded convenience sample of patients with signs and symptoms of ADHF who underwent measurement of Zo with concomitant chest radiography. Attending physicians blinded to the Zo values interpreted the radiographs, categorizing the severity of pulmonary edema as normal (NL), cephalization (CZ), interstitial edema (IE), or alveolar edema (AE). Intergroup comparisons were analyzed with a 2-way analysis of variance (ANOVA), with P < .05 considered statistically significant and reported using 95% confidence intervals (CIs). SETTING: We enrolled patients (> or =18 years) presenting to a tertiary care medical center ED with signs and symptoms consistent with ADHF. RESULTS: A total of 203 patients were enrolled, with 27 (14%) excluded because of coexisting pulmonary diseases. The mean Zo values were inversely related to the 4 varying degrees of radiographic pulmonary vascular congestion as follows: NL, 25.6 (95% CI, 22.9-28.3); CZ, 20.8 (95% CI, 18.1 23.5); IE, 18.0 (95% CI, 16.3-19.7); and with AE, 14.5 (95% CI, 12.8-16.2) (ANOVA, P < .04). A Zo less than 19.0 ohms had 90% sensitivity and 94% specificity (likelihood ratio [LR], - 0.1; LR + 15) for identifying radiographic findings consistent with pulmonary edema. Females had an increased mean Zo value compared to males (P < .03). CONCLUSION: The Zo value obtained via thoracic bioimpedance monitoring accurately predicts the presence and severity of pulmonary edema found on initial chest radiograph in patients suspected of ADHF. PMID- 19683104 TI - Body surface mapping vs 12-lead electrocardiography to detect ST-elevation myocardial infarction. AB - A prospective, multicenter trial was conducted in patients with nontraumatic chest pain in 4 hospitals to determine whether an 80-lead body surface map electrocardiogram system (80-lead BSM ECG) improves detection of ST-segment elevation in acute myocardial infarction (STEMI) compared with a standard 12-lead electrocardiogram (ECG) in an emergency department (ED) setting. A trained ED or cardiology staff member (technician or nurse) recorded a 12-lead ECG and 80-lead BSM ECG from each subject at initial presentation. Serial biomarkers (total creatine kinase [CK], CK-MB, and/or troponin) were obtained according to individual hospital practice. Of the 647 patients evaluated, 589 had available biomarkers results. Eighty-lead BSM ECG improved detection of biomarker-confirmed STEMI compared with the 12-lead ECG for CK-MB-defined STEMI (100% vs 72.7%, P = .031; n = 364) or troponin-defined STEMI (92.9% vs 60.7%, P = .022; n = 225). Specificity for STEMI was high (range, 94.9%-97.1%) with no significant difference between 80-lead BSM ECG and 12-lead ECG. Right ventricular involvement complicating inferior STEMI was detected by 80-lead BSM ECG in 2 (22%) of 9 patients with CK-MB-defined MI and in 2 (22%) of 9 patients with troponin-defined MI. The infarct location missed most commonly on 12-lead ECG but detected by 80 lead BSM ECG was inferoposterior MI. We conclude that BSM using 80-lead BSM ECG is more sensitive for detection of STEMI than 12-lead ECG, while retaining similar specificity. PMID- 19683105 TI - The worsening of ED on-call coverage in California: 6-year trend. AB - To reassess problems with on-call physician coverage in California, we repeated our anonymous 2000 survey of the California chapter of the American College of Emergency Physicians. Physicians responded from 77.4% of California emergency departments (EDs), 51.0% of ED directors, and 34% of those surveyed. Of 21 specialties, on-call availability worsened since 2000 for 9, was unchanged for 11, and improved for 1. Of ED directors, 54% report medical staff rules require on-call duty, down from 72% in 2000. Hospitals have increased specialist on-call payments (from 21% to 35%, with 75% paying at least one specialty). Most emergency physicians (80.3%) report insurance status negatively affects on-call physician responsiveness, up from 42% in 2000. Emergency departments with predominantely minority or uninsured patients had fewer specialists and more trouble accessing them. Insurance status has a major negative effect on ED consultation and follow-up care. The on-call situation in California has worsened substantially in 6 years. PMID- 19683106 TI - Evaluation of pelvic fracture stability and the need for angioembolization: pelvic instabilities on plain film have an increased probability of requiring angioembolization. AB - BACKGROUND: The use of pelvic x-rays (PXRs) as an early diagnostic adjunct in the evaluation of blunt trauma patients has become widely accepted, and computed tomographic (CT) scanning is now used universally in the assessment of abdominal and pelvic injuries. In this study, we have attempted to identify patients with pelvic fractures who might be at risk for vessel hemorrhage and determine if early angioembolization was required in these patients. MATERIAL AND METHODS: We retrospectively reviewed patients who presented with pelvic fractures from June 2005 to August 2007. Both PXRs and CT scans were reviewed. Patients who presented with bleeding due to other associated injuries or who did not receive a CT scan were excluded. Patients with either initial hemodynamic instability or contrast extravasation on enhanced CT scan underwent angioembolization. Patient demographics, Injury Severity Score, the amount of blood transfused, and the relationship between the fracture pattern and angioembolization were analyzed. RESULTS: A total of 54 patients were enrolled. A diagnosis of an unstable pelvic fracture on PXR was associated with a higher probability of angioembolization. Seven patients received incompatible diagnoses from the PXR and CT scan; these patients received larger amounts of transfused blood and demonstrated an increased need for angioembolization. CONCLUSIONS: Although CT scan is more sensitive in the identification of acetabular or small pelvic fractures, PXR is sufficient for the early evaluation of pelvic fracture stability. Based on the current series, early angioembolization is suggested for patients with an initial diagnosis of an unstable pelvic fracture. In addition, patients receiving large amounts of transfused blood are more likely to require early angioembolization. PMID- 19683107 TI - Admission and length of stay due to painful vasoocclusive crisis in children. AB - PURPOSE: To identify demographic, clinical, and laboratory characteristics associated with admission and a longer length of stay (LOS) due to vasoocclusive crisis (VOC) in children with sickle cell disease (SCD). METHODS: Retrospective chart review at a large tertiary pediatric center. Patients younger than 18 years with VOC due to SCD presenting to the emergency department were included. We performed multivariate regression analyses to predict characteristics associated with admission and LOS of 4 days or more. RESULTS: A total of 428 visits for VOC were documented in 2005 to 2006. In a multivariate regression analysis higher pain score at triage (P < .001), older age (P = .04), and increased systolic blood pressure (P = .02) were predictors of admission. Higher pain score at triage (P = .046), older age (P = .002), increased polymorphonuclear count (P = .02), and homozygous SCD type (P = .03) were associated with prolonged hospital LOS. CONCLUSION: These characteristics will help healthcare providers predict and plan admission and management of children with SCD. PMID- 19683108 TI - Emergency endoscopy for upper gastrointestinal bleeding in patients with coronary artery disease. AB - BACKGROUND: Endoscopy is useful for diagnosis and treatment of upper gastrointestinal bleeding (UGIB). However, both endoscopy and UGIB may compromise the cardiovascular function. The present study is to investigate the cardiovascular responses of emergency endoscopy for patients with UGIB and stable coronary artery disease (CAD). METHODS: Consecutive 50 patients with known CAD and 50 patients without CAD history (non-CAD group) in whom emergency endoscopy was requested for UGIB were prospectively enrolled. All patients received ambulatory electrocardiographic monitoring before, during, and after endoscopies. Cardiac indices including supraventricular and ventricular arrhythmia, ST ischemic change, and autonomic nervous function evaluated by heart rate variability were compared. RESULTS: All patients in both groups had successful primary hemostasis, and peptic ulcer bleeding was the main etiology (82%). Compared with the non-CAD group, patients with CAD had a significantly higher incidence (42% vs 16%, P = .004) and frequency (1.19 vs 0.12 events per minute, P = .003) of ventricular arrhythmias during endoscopy. Nine patients with CAD and 1 patient without CAD had ischemic ST changes (P = .016). Comorbidity with congestive heart failure was not only associated with a higher frequency (P = .02) but also a more severe fluctuation (P = .002) of ventricular arrhythmia. None in both groups had angina or MI before, during, or after endoscopy. Heart rate variability did not show a difference. CONCLUSIONS: Ventricular arrhythmias and myocardial ischemia, although mostly subclinical, were common in patients with stable CAD undergoing emergent endoscopy for UGIB, especially in those with concomitant congestive heart failure. PMID- 19683109 TI - Improving length-based weight estimates by adding a body habitus (obesity) icon. AB - BACKGROUND: Length-based dosing systems reduce errors associated with resuscitation drug dosing. Obese and thin children of the same length are dosed the same despite their different weights. METHODS: Length (height) and weight were measured in children after a body habitus icon assignment. Within each body habitus group, regression analysis was performed to generate a weight-estimation formula using body habitus and length (BHL). This BHL method was compared to the Broselow tape (BT). RESULTS: Height and weight data were plotted to obtain visual scattergrams. Logarithmic regression yielded higher correlation coefficients than standard linear regression. Within body habitus groups, BHL epinephrine dose estimates were more accurate than BT dose estimates using 0.01 mg/kg as a dosing standard. CONCLUSIONS: Adding body habitus information to the patient's length results in a more accurate weight estimate than length alone in children. The accuracy improvement is greater in children 3 years and older as compared to younger children. PMID- 19683110 TI - Performance of a body surface mapping system using emergency physician real-time interpretation. AB - OBJECTIVE: We compared the performance characteristics of the 12-lead electrocardiography (ECG) with body surface mapping (BSM) in patients presenting for evaluation of symptoms suggestive of acute coronary syndromes. METHODS: The diagnostic test characteristics (sensitivity, specificity, likelihood ratios, and predictive values) for 12-lead ECG and BSM were computed using 3 different criterion standards. RESULTS: Of the 150 patients enrolled, 19 were positive for acute coronary syndromes using the criterion standard of cardiac troponin T >0.1 ng/mL, percutaneous coronary intervention, more than 70% stenosis, abnormal noninvasive testing, and coronary artery bypass graft. Changes not known to be old on ECG and BSM had sensitivities of 10.5 (95% confidence interval [CI(95)], 1.8-34.5) and 15.8 (CI(95), 4.2-40.5), and specificities of 90.1 (CI(95), 83.3 94.4) and 86.3 (CI(95), 78.9-91.4), respectively. CONCLUSION: In this emergency department population, both the BSM and the 12-lead ECG exhibited similar test characteristics. PMID- 19683111 TI - A comparison of 2 types of chest compressions in a porcine model of cardiac arrest. AB - OBJECTIVE: Chest compressions performed by some medical workers are of poor quality, which are too few and shallow with incomplete release. This study was designed to compare the effects of these clinical quality chest compressions with standard manual chest compressions in a porcine model of cardiac arrest. METHODS: Ventricular fibrillation was induced in 18 pigs by programed electrical stimulation. Then, 40 mg methylene blue was injected into right atrium after 4 minutes of untreated ventricular fibrillation (VF), followed by cardiopulmonary resuscitation for 9 minutes. Defibrillation was attempted at 13 minutes of cardiac arrest. Animals of no restoration of spontaneous circulation after 4 times of defibrillations were announced dead and dissected immediately to observe the cerebral perfusion with methylene blue coloration. Resuscitated animals were executed to remove the tissues of pallium, cardiac muscle, kidney, and liver for histopathology after evaluating a porcine Cerebral Performance Category score at 24 hours after cardiac arrest. All animals were randomized to the following 2 groups: (1) standard manual chest compressions group (n = 9)-chest compression rates were kept at 100 +/- 5 cpm and compression depth at 50 +/- 1 mm with complete release by Heartstart MRx Monitor; (2) clinical quality chest compressions group (n = 9)-chest compression rates were kept at 80 +/- 5 cpm and compression depth at 37 +/- 1 mm with incomplete release. RESULTS: Compared with clinical quality chest compressions, standard manual chest compressions produced greater restoration of spontaneous circulation, neurologically normal 24-hour survival, and histopathologic findings. CONCLUSIONS: High-quality chest compressions improve outcomes of resuscitation, especially postresuscitation brain damage. PMID- 19683112 TI - The relationship between paramedics' level of education and degree of commitment. AB - INTRODUCTION: Emergency medical services (EMS) personnel attrition is a serious concern. Two fundamental psychological constructs linked to attrition are organizational and occupational commitment. OBJECTIVE: To determine if there is a relationship between a paramedic's degree of occupational/organizational commitment and the following: (1) levels of education and (2) type of employment. METHODS: This was a cross-sectional study of paramedics in 6 states that require continued paramedic national registration. The data collection instrument consisted of demographic and occupational and organizational commitment sections. For level of education, the primary independent variable, each subject was placed into 1 of 3 groups: (1) certificate, (2) associate's or bachelor's degree in EMS (degree), and (3) paramedic certificate or degree with a non-EMS postbaccalaureate degree. Type of employment (fire based vs non-fire based) was also used as an independent variable. Organizational and occupational commitment was measured using validated scales for each. Analysis of variance was used for the comparisons between levels of each of the independent variables. A P < .05 was considered significant. RESULTS: For occupational commitment, the participants with certificate level of education had a significantly higher score (88.9) than did those with either the degree (83.6) or postbaccalaureate (80.9) level of education. There were no significant differences for total organizational commitment. There were also no overall differences in occupational and organizational commitment between fire- and non-fire-based employees. CONCLUSION: Paramedic occupational commitment shows a statistically significant decrease with increased level of education. Factors associated with commitment of more highly educated paramedics need to be explored. PMID- 19683114 TI - Expedited admission of patients decreases duration of mechanical ventilation and shortens ICU stay. AB - BACKGROUND: To determine if expedited admission (<2 hours) of critically ill patients requiring intubation and mechanical ventilation from the emergency department (ED) to the intensive care unit (ICU) decreases ICU and hospital length of stay. METHODS: Patients with respiratory failure that required intubation and mechanical ventilation who were admitted to the hospital between June 2004 and May 2006 were retrospectively identified from the Project IMPACT database. Patients were divided into 2 groups based on ED length of stay: expedited (<2 hours) or nonexpedited (>2 hours). RESULTS: The expedited (n = 12) and nonexpedited (n = 66) groups were comparable in demographics, medical conditions, and disease severity. Mean duration of mechanical ventilation was significantly shorter in the expedited group (28.4 hours vs 67.9 hours; P = .0431), as was mean ICU length of stay (2.4 days vs 4.9 days; P = .0209). Length of hospital stay tended to be shorter for the patients in the expedited group (6.8 days vs 8.9 days; P = .0609). CONCLUSIONS: Expedited admission (<2 hours) of critically ill patients requiring intubation and mechanical ventilation from the ED to the ICU was associated with shorter durations of mechanical ventilation and ICU length of stay, suggesting that prompt ICU admission results in improved use of resources. PMID- 19683113 TI - Elevated serum S100B protein and neuron-specific enolase levels in carbon monoxide poisoning. AB - OBJECTIVE: Carbon monoxide (CO) poisoning causes cerebral and generalized hypoxia. This study aimed to assess the possible use of serum glial marker S100B protein and neuron-specific enolase (NSE) as biochemical markers of hypoxic brain damage in acute CO poisoning. METHODS: Patients with acute CO poisoning admitted to the ED of 2 training hospitals (Ankara, Turkey) were included in this cross sectional study. Serum levels of S100B and NSE were measured on admission. The patients were divided into 2 groups (unconscious and conscious). Twenty healthy adults were included in the study to serve as controls. RESULTS: A total of 70 patients poisoned by CO (mean age +/- SD, 36.6 +/- 16.3 years; 64.3% women) were enrolled. Although S100B concentrations were higher in patients than in the control group (P = .018), no significant difference was determined between patient and control groups with respect to NSE concentrations (P = .801). A positive correlation was noted between levels of S100B and NSE (r = 0.388; P = .001). The S100B and NSE values were higher in unconscious patients than in the control group (P = .002 and P = .013, respectively). Furthermore, S100B and NSE values were higher in unconscious vs unconscious patients (P = .047 and P = .005, respectively). CONCLUSION: Elevated serum S100B and NSE levels were associated with loss of consciousness in CO poisoning in this series of patients. Serum S100B and NSE may be useful markers in the assessment of clinical status in CO poisoning. PMID- 19683115 TI - A comparison of the palatability of flavored oral contrasts. AB - OBJECTIVE: The aim of this study was to compare the taste of computed tomography (CT) oral contrast diluted with various flavored drinks. METHODS: We performed a prospective, blinded, controlled trial in healthy adult volunteers. Subjects were assigned to ingest four 250-mL aliquots of oral contrast media diluted in water, Crystal Light Lemonade (Kraft Food, Northfield, Ill), Tropical Punch Kool-Aid (Kraft Food), and Tropicana orange juice (Pepsi Bottling Company, Sommers, NY) in random order; and the taste of the solution was measured with a 100-mm visual analogue scale and 5-point Likert scale from very worst to best. Between-group comparison of the taste scores was performed with repeated-measures analysis of variance and pairwise t tests. The study had 80% power to detect an effect size 0.75 SDs. RESULTS: There were 23 subjects; mean (SD) age was 33 (7.7) and 30% were female. The mean (SD) taste scores were water 12 (5), lemonade 37 (21), Kool Aid 44 (20), and orange juice 40 (20) (P < .05). The proportion of subjects completely ingesting the contrast in water (65%) was significantly less than that with other 3 study solutions (100% each, P < .001). CONCLUSIONS: Dilution of oral contrast media with lemonade, fruit punch, or orange juice is tastier than with water. The choice of the specific juice used to dilute the oral contrast should be individualized based on patient preferences and availability. PMID- 19683116 TI - Correlation of sonographic measurements of the internal jugular vein with central venous pressure. AB - Determination of volume status is crucial in treating acutely ill patients. This study examined bedside ultrasonography of the internal jugular vein (IJV) to predict central venous pressure (CVP). Ultrasonography was performed on 34 nonventilated patients with monitored CVPs. The IJV was measured during the respiratory cycle and with the patient in different positions. Mean IJV diameter in patients with CVP less than 10 cm H2O was 7.0 mm (95% confidence interval [CI], 5.7-8.3) vs 12.5 mm (95% CI, 11.2-13.8) in patients with CVP of 10 cm H2O and greater. Measurement of end expiratory diameter with the patient supine had the highest correlation coefficient: 0.82 (95% CI). There was strong agreement among ultrasonographers: correlation coefficient, 0.92 (95% CI). This pilot study shows promise that ultrasonography of the IJV can be a noninvasive tool to predict CVP. Measurement of end expiratory diameter in supine patients exhibited a high correlation to CVP. PMID- 19683117 TI - Is female sex associated with ED delays to diagnosis of appendicitis in the computed tomography era? AB - BACKGROUND: Historically, females had delays to definitive diagnosis of appendicitis when compared to males. In this current millennium, appendicitis is now most commonly diagnosed by computed tomography (CT) in the emergency department (ED) rather than at surgery. OBJECTIVE: The aim of the study was to assess if female gender is still associated with delays to diagnosis of appendicitis in the CT era. METHODS: A retrospective cohort analysis of adult patients with appendicitis at a university teaching hospital ED was conducted. Inclusion criteria was age of more than 18 years and an International Classification of Diseases, Ninth Revision (ICD-9), diagnosis of appendicitis. Patients were excluded from analysis if they were pregnant, no CT scan was obtained in the ED, or had incomplete outcome data. RESULTS: One hundred thirty seven patients met inclusion criteria; 65 female, 72 males. Time from triage to CT order was 138 minutes in females and 95 minutes in males (P = .0012). Time from initial physician evaluation to CT order was 45 minutes in females and 28 minutes in males (P = .0012). Nonclassic symptoms were more common in females and pelvic evaluation did not delay the CT order. CONCLUSION: Female gender is still associated with delays to CT acquisition and diagnosis of appendicitis. PMID- 19683118 TI - The usefulness of the semiquantitative procalcitonin test kit as a guideline for starting antibiotic administration. AB - OBJECTIVES: The Surviving Sepsis Campaign has recommended that antibiotic therapy should be started within the first hour of recognizing severe sepsis. Procalcitonin has recently been proposed as a biomarker of bacterial infection, although the quantitative procalcitonin assay is often time consuming, and it is not always available in many emergency departments (EDs). Our aim is to evaluate usefulness of the semiquantitative procalcitonin fast kit as a guideline for starting antibiotic administration for patients with severe sepsis or septic shock that requires prompt antibiotic therapy in the ED. METHODS: We include those patients who were admitted to the ED and who were suspected of having infection. The procalcitonin concentration was determined by semiquantitative PCT Q strips, and the points of the severity scoring system were calculated. The receiver operating characteristic curve was used to assess the diagnostic value of the PCT-Q strips to predict severe sepsis or septic shock. RESULTS: Of the 80 recruited patients, 33 patients were categorized as having severe sepsis or septic shock according to the definition. At a procalcitonin cutoff level of 2 ng/mL or greater, the sensitivity of the PCT-Q for detecting severe sepsis or septic shock was 93.94% and the specificity was 87.23. The receiver operating characteristic curve for PCT-Q to predict severe sepsis or septic shock had an area under the curve of 0.916. CONCLUSION: PCT-Q is probably a fast, useful method for detecting severe sepsis in the ED, and it can be used as a guideline for antibiotic treatment. PMID- 19683119 TI - The Rusch ViewMax vs Macintosh laryngoscopy in human cadavers: impoved vocal cord visualization? AB - OBJECTIVES: Adequate vocal cord visualization via laryngoscopy is a key component of successful endotracheal intubation. Several tools exist to facilitate laryngoscopy in difficult airways. We compared one such device, the Rusch "ViewMax" (Rusch, Duluth, Ga), to a standard Macintosh laryngoscope blade (Heine USA Ltd, Dover, NH) using human cadaver models. The purpose of this study was to determine if the ViewMax improved vocal cord visualization. METHODS: Emergency medicine residents and faculty (N = 26) attempted vocal cord visualization with both ViewMax and Macintosh laryngoscope blades for each of 6 cadaver airways at an airway laboratory training exercise. Percentage of glottic opening (POGO) score was estimated for each laryngoscopy attempt. Data were analyzed by nonparametric statistics. RESULTS: Of 6 cadaver airways, 4 had median POGO score estimates of 100% for both ViewMax and Macintosh laryngoscope blades. Two cadavers had median POGO estimates of less than 50% for both blades. No difference was found in POGO score estimates between the ViewMax and the Macintosh blades in any of the 6 cadavers including those with more difficult vocal cord visualization (P = .27, .35, .61, .40, .39, .31). CONCLUSION: The Rusch "ViewMax" was not shown to improve vocal cord visualization over standard Macintosh blade laryngoscopy in these cadaver models. Further study with cadaver models known to have more difficult airways may improve power to detect a small difference in vocal cord visualization, though the clinical relevance of any slight difference remains uncertain. PMID- 19683120 TI - HIV testing in US EDs, 1993-2004. AB - OBJECTIVES: The aims of the study were to (1) estimate the incidence rates (IRs) of human immunodeficiency virus (HIV) testing among 13-year-old to 64-year-old patients in US emergency departments (EDs); (2) determine ED compliance with Centers for Disease Control and Prevention (CDC) recommendations for HIV testing for patients with nonsexual blood or body fluid exposures, sexually transmitted diseases (STDs), and sexual assaults; and (3) ascertain if HIV testing in EDs varies by patient demographic characteristics. METHODS: The ED visits from the National Hospital Ambulatory Medical Care Survey databases (1993-2004) were analyzed. Visits for nonsexual blood or body fluid exposures, STDs, and sexual assaults were identified using diagnosis and cause codes. Incidence rates for HIV testing were estimated by year. Odds ratios (ORs) with 95% confidence intervals were estimated from multivariable logistic regression models using HIV testing as the outcome and demographic characteristics as covariates. RESULTS: The average IR of HIV testing for 13-year-old to 64-year-old patients from 1993 to 2004 was 0.31%. Of all patients, 35.1% with nonsexual blood or body fluid exposures, 20.4% with sexual assaults, and 2.6% with STDs were tested for HIV. The HIV testing was more frequent among Hispanics (OR, 1.39 [1.06-1.81]), blacks (OR, 1.52 [1.19 1.94]), patients with Medicaid (OR, 2.35 [1.81-3.03]), Medicare (OR, 1.95 [1.20 3.16]), and self-pay/no charge/other type of insurance (OR, 1.74 [1.35-2.23]), and those visiting EDs in the northeastern United States (OR, 1.57 [1.04-2.38]). CONCLUSIONS: The HIV testing rates are low in US EDs and have changed little for a 12-year period. Compliance with CDC recommendations for HIV testing is poor and not in accordance with risk for infection. Hispanics, blacks, and those without private health care insurance are being tested more frequently than other ED patients. PMID- 19683122 TI - Preexcitation syndromes: diagnostic consideration in the ED. AB - Preexcitation syndromes are a common cause of paroxysmal tachycardias presenting to the ED. Emergency physicians should be familiar with the common electrocardiographic manifestations of preexcitation, particularly the Wolff Parkinson-White abnormality, as these conditions require specific therapeutic management. This article reviews the pathophysiology of preexcitation, along with the electrocardiographic findings of Wolff-Parkinson-White and its associated tachyarrhythmias. PMID- 19683121 TI - Urine dipstick testing to rule out rhabdomyolysis in patients with suspected heat injury. AB - BACKGROUND: Heat injury is a common, potentially life-threatening medical condition. In austere or mass-casualty conditions an easy to use, sensitive screening test could be a valuable tool to care providers and evacuation planners. OBJECTIVE: The objective of the study was to determine if a simple urine dipstick test for blood is sensitive for detection of rhabdomyolysis in the suspected heat injury patient. MATERIAL AND METHODS: A convenience sample of patients presenting to a military community hospital Emergency Department during summer months with a presenting complaint consistent with suspected heat injury had urine dipstick testing performed for blood and compared with the results of formal urinalysis and serum creatine kinase. RESULTS: 60 patients were enrolled in the study, seven had creatine kinase levels greater than 1000 U/L, 14 had levels greater than 500 U/L, and 26 had levels greater than 250 U/L. Using 1000 U/L, urine dipstick testing had a sensitivity of 14% and a specificity of 85%. CONCLUSIONS: Urine dipstick testing for blood is not a useful screening test for rhabdomyolysis in patients suspected to have significant heat injury. PMID- 19683124 TI - Rifampin-containing regimens for community-associated skin infection: a hazard without known benefit. PMID- 19683123 TI - The use of exercise stress testing for the management of low-risk patients with chest pain. AB - OBJECTIVE: This retrospective study assessed the contribution of exercise stress testing (EST) in the evaluation of patients with low risk for coronary heart disease who presented to the emergency department (ED) with chest pain. BASIC PROCEDURES: The study included 175 patients who presented to the ED with chest pain and underwent EST between January 1, 2005, and November 30, 2006. MAIN FINDINGS: After the EST, 113 patients were discharged, and 62 were admitted. Exercise stress testing's positive predictive value for coronary artery disease among admitted patients was 35.7%, and sensitivity was 95.2%. Exercise stress testing's negative predictive value among discharged patients was 99.1%. None of the 113 discharged patients returned to the ED for cardiac reasons during the 30 day follow-up period. PRINCIPAL CONCLUSION: A chest pain unit or a parallel facility for evaluating patients with chest pain and with low risk for active coronary disease is necessary for detecting low-risk patients who eventually need cardiac intervention. PMID- 19683125 TI - Definition of cyanosis. PMID- 19683126 TI - Bone pain as an atypical early manifestation of acute myocardial infarction. PMID- 19683127 TI - Purple urine bag syndrome, not always a benign process. PMID- 19683128 TI - T-wave inversion: cardiac memory or myocardial ischemia? AB - This article presents a case report of a 74-year-old man with T-wave inversion (TwI) in atrial fibrillation noted during routine pacemaker interrogation. The patient was seen for routine pacemaker interrogation, at which time he was noted to have underlying atrial fibrillation. A12-lead electrocardiogram of the atrial fibrillation revealed significant TwIs. He was subsequently worked up for myocardial ischemia and was found to have a moderate-sized,moderate-degree inferior wall myocardial perfusion defect. He was subsequently referred for a cardiac catheterization. The cardiac catheterization revealed nonobstructive coronary artery disease. The follow-up electrocardiogram revealed persistent but attenuated TwI.The TwIs were attributed to cardiac memory, a common but infrequently recognized phenomenon of which many clinical practitioners are unaware. Cardiac memory is due to the T wave tracking the preceding abnormal QRS complex and can be induced by right ventricular pacing or arrhythmias. PMID- 19683129 TI - Rupture of splenic artery aneurysm during early pregnancy: a rare and catastrophic event. AB - Spontaneous rupture of splenic artery aneurysm during pregnancy is a rare, life threatening event with a catastrophic prognosis. Splenic artery aneurysm, known to be more frequent in women, especially among multiparous ones, is generally asymptomatic until rupture. Because of increased blood flow and hormonal modifications, this rupture occurs frequently during pregnancy, most often at the end of the third trimester. We present the case of a second parous woman in early pregnancy with a rupture of splenic artery aneurysm initially diagnosed as a complicated ectopic pregnancy, which profoundly modifies surgical treatment. Through this case report, we want to draw attention to the fact that even if complicated ectopic pregnancy is much more frequent, ruptured splenic artery aneurysm (SAA) needs to be considered as a part of differential diagnosis of hemoperitoneum during the first trimester as well. PMID- 19683130 TI - A case of rectus sheath hematoma caused by yoga exercise. AB - Rectus sheath hematoma is a relatively rare condition but a significant cause of acute abdominal pain. The acute abdominal pain associated with rectus sheath hematoma can mimic several conditions, such as appendicitis, intestinal problems (obstruction, perforation, hernia, malignancy), and gynecologic disease (ovarian cyst, torsion, tubo-ovarian abscess, ectopic pregnancy). Correct diagnosis and subsequent treatment depends on careful history taking and appropriate use of diagnostic tools to avoid unnecessary laparotomy. We report a case of rectus sheath hematoma due to noncontact strenuous exercise, yoga, that mimicked gynecologic diseases. PMID- 19683131 TI - Acute myocardial infarction with cardiogenic shock in a patient with acute aortic dissection. AB - Diagnosing acute Stanford type A aortic dissection with the uncommon involvement of the left main coronary artery(LMCA) remains challenging for the emergency physician because it can resemble acute myocardial infarction with cardiogenic shock. The following case report illustrate this infrequent but critical situation. A 52-year-old woman with a history of hypertension awakened with acute retrosternal chest pain accompanied by nausea and vomiting. She was referred to our hospital for primary coronary intervention because of acute myocardial infarction with cardiogenic shock. Coronary angiography indeed revealed LMCA occlusion. Subsequently successful percutaneous coronary intervention with stent implantation was performed, followed by immediate clinical improvement of the patient. Soon after admission at the coronary care unit, severe chest pain, hypotension, and electrocardiographic signs of diffuse myocardial ischemia relapsed. Control coronary angiography,however, showed no in-stent thrombosis. Review of clinical examination revealed an aortic regurgitation murmur. Because of this dynamic pattern of (1) signs of acute myocardial ischemia, (2) relapse of hemodynamic collapse, and (3) unaltered control coronary angiography together with the confirmed aortic regurgitation at transthoracic echocardiography, the patient was suspected of having aortic dissection. Transesophageal echocardiography revealed Stanford type A aortic dissection with severe eccentric aortic regurgitation and no pericardial effusion. Emergent valve-sparing aortic replacement was performed. The patient recovered completely. In this case, the lifesaving element was primary coronary intervention with stenting of the LMCA preventing extensive myocardial damage followed by a surgical correction of the aorta. PMID- 19683132 TI - Thirteen-week cornual ectopic pregnancy. AB - We present the unusual case of a 13-week cornual ectopic pregnancy with fetal movement. The patient presented with a protuberant abdomen, intoxicated with cocaine and alcohol, and in hemodynamic compromise. Bedside emergency ultrasonography led to rapid diagnosis and treatment. PMID- 19683133 TI - Acute inflammatory demyelinating polyneuropathy as the initial presentation of lupus. AB - A 28-year-old woman presented at the emergency department (ED) with acute bilateral leg weakness and lost the ability to walk 1 day after noticing bilateral leg numbness and intermittent pleuritic pain for 3 weeks. Physical examination revealed bilateral decreased muscle strength with score 4 symmetrically in the legs and decreased deep tendon reflex. Laboratory tests indicated lymphopenia but no other grossly remarkable findings. Cerebrospinal fluid analysis demonstrated albuminocytologic dissociation. Electrophysiologic survey also indicated sensory-motor demyelinating polyneuropathy. The clinical picture was compatible with acute inflammatory demyelinating polyneuropathy (AIDP), the most prevalent form of Guillain-Barre syndrome (GBS). Plasma exchange was immediately arranged. After serial examination, pleuritic pain, persistent lymphopenia, positive immunologic findings of anti-DNA, and antinuclear antibodies led to a diagnosis of systemic lupus erythematosus (SLE). Prednisolone was added along with plasma exchange. The patient was able to walk after 2 weeks of therapy. Acute inflammatory demyelinating polyneuropathy presenting as the initial manifestation of SLE is rather rare. The precise mechanism of SLE-related AIDP remains unclear but is probably immune related. Although steroids are not recommended in the management of AIDP or GBS, patients with SLE-related AIDP may benefit from steroid therapy. This case highlights that early initiation of evaluation for SLE by ED physicians may facilitate correct diagnosis and better outcomes in patients presenting with GBS or AIDP in the ED. PMID- 19683135 TI - Bilateral peroneal compartment syndrome after horse riding. AB - A healthy 20-year-old woman developed acute ischemia of the lateral compartment of both calves shortly after a 30-minute horse ride. On one side, she developed compartment syndrome with resultant complete myonecrosis of the compartment, whereas on the other side, there was spontaneous resolution. To our knowledge, this is the first report of bilateral lateral compartment ischemia after horse riding. Atraumatic compartment syndrome is a rare entity and is often missed at initial presentation. We discuss aspects of her management together with a review of the literature. Late fasciotomy and exploration may be beneficial in decompressing the deep peroneal nerve in peroneal compartment syndrome. Awareness of atraumatic compartment syndrome is important in any case of limb pain and swelling. PMID- 19683134 TI - Adenosine-induced cardiopulmonary arrest in a patient with paroxysmal supraventricular tachycardia. AB - Adenosine is commonly used for the chemical termination of supraventricular tachycardia. In addition, even when it is ineffective as an agent of chemical cardioversion, it may slow the cardiac rate to allow an analysis of the underlying rhythm. Common adverse effects include facial flushing, shortness of breath, and chest pain. Major contraindications include heart blocks and known adenosine hypersensitivity. This case report illustrates an episode of cardiopulmonary arrest after adenosine administration and, to the authors' knowledge, is the first occurrence reported in the literature. PMID- 19683136 TI - Spinal epidural hematoma after stretch exercise: a case report. AB - Spinal epidural hematoma (SEH) is a rare but serious condition. Common causes of SEH include spinal fracture, spinal trauma, and invasive spinal procedures such as lumbar puncture, epidural anesthesia, or myelography. A few previous reports have suggested that SEH could be caused by minor triggers such as spinal manipulation therapy or spontaneously in rare cases. A 60-year-old man, immediately after stretch exercise, developed severe back cervicodynia and rapidly progressive weakness of left arm from SEH. He was treated by decompression laminectomy and evacuation of the hematoma. However, the upper extremity weakness did not completely resolve. The standard treatment remains timely surgical decompression and evacuation of the hematoma. Thus, early diagnosis and treatment can confer a significant prognostic advantage to patients with SEH. Based on the finding that SEH was caused by a mild mechanical trigger and neurologic deficits remained in this patient, this report highlights its diagnostic difficulty as well as the importance of rapid treatment. Emergency physicians should consider SEH among the differential diagnosis in patients with sudden-onset back pain with symptoms and signs of spinal cord compression. PMID- 19683137 TI - Entire pneumorrhachis due to isolated head trauma. AB - Air contained within the spinal canal is termed pneumorrhachis, and very rarely, traumatic pneumocephalus can cause pneumorrhachis. Pneumorrhachis can be classified as epidural or subarachnoid space pneumorrhachis. PMID- 19683138 TI - Trastuzumab (Herceptin)-associated cardiomyopathy presented as new onset of complete left bundle-branch block mimicking acute coronary syndrome: a case report and literature review. AB - Trastuzumab (Herceptin) is well documented in reducing suffering and mortality from breast cancer. The clinically most important side effect of Herceptin is cardiotoxicity, which is reported in 2.6% to 4.5% of patients receiving trastuzumab alone and in as many as 27% of patients when trastuzumab is combined with an anthracycline in metastatic disease. We reported the case of a 50-year old woman who presented to our emergency department (ED) because of chest pain and shortness of breath. On physical examination, holosystolic murmur over apex could be heard. Pulmonary and abdominal examinations were unremarkable. Twelve lead electrocardiography showed sinus tachycardia and new onset of complete left bundle-branch block. Emergent transthoracic echocardiography revealed generalized hypokinesia of left ventricle and akinesia over interventricular septum and apex. She subsequently underwent immediate coronary angiography that revealed normal coronary angiography, and left ventriculogram revealed generalized hypokinesia with severe left ventricle dysfunction with ejection fraction of 33%. During right heart catheterization and endomyocardial biopsy, cardiac tamponade developed and was successfully relieved by pericardial window. She was discharged event-free 3 weeks later with conservative treatment. Although new onset of complete left bundle-branch block in a patient with chest pain may be acute coronary syndrome, careful review of medicine history is mandatory to avoid unnecessary procedure and complications. PMID- 19683139 TI - Multiply organ dysfunction syndrome due to tramadol intoxication alone. AB - Tramadol is a synthetic, centrally acting analgesic for the treatment of moderate to severe pain. It is widely used because of its efficacy and safety. A normal dose tramadol has less adverse effects than other opioids. Multiply organ dysfunction syndrome due to tramadol intoxication is rare.We present a case of 19 year-old male patient who had multiply organ dysfunction syndrome due to oral tramadol alone. With a history of tramadol abuse for 6 months, the patient was found unconsciousness in bed 8 hours before hospitalization. A toxicologic analysis showed a tramadol blood concentration of 9.5 mg/L without toxic levels of other drugs. The patient developed deep coma, acute respiratory distress syndrome, hepatic and renal dysfunction,and shock in the first 24 hours after admission. With mechanical ventilation, hemoperfusion, and other supportive therapies, his overall status gradually improved and he was discharged 23 days later. PMID- 19683140 TI - Refractive surgery experience for the ophthalmology resident: an update. PMID- 19683141 TI - In situ nuclear disassembly: efficient phacoemulsification without nuclear rotation using lateral sweep sculpting and in situ cracking techniques. AB - We describe an in situ nuclear disassembly technique using a simplified form of sculpting and cracking, enhanced by phacoemulsification technology with a Kelman style tip. The technique does not require mobilization or rotation of the nucleus within the capsular bag, and hydrodissection is optional. The technique requires a Kelman-style curved tip and Kuglen hook and takes advantage of the versatility of the curved phacoemulsification needle for sculpting and cracking. Since in situ nuclear disassembly requires no rotation of the nucleus for extraction, it is useful in cases in which zonules are compromised or the surgeon suspects posterior lenticonus and the potential for loss of integrity of the posterior capsule. PMID- 19683142 TI - Sutured intraocular lens placement in aphakic post-vitrectomy eyes via small incision surgery. AB - A technique for placing a sutured intraocular lens (IOL) in post-vitrectomy eyes is described. A foldable acrylic IOL (Akreos) is sutured to the sclera with 4 point fixation via a standard cataract incision, avoiding the large incision needed for nonfoldable IOLs, which would represent extra risk in eyes with no internal support. Postoperative results in 2 cases were significant for quick recovery and minimal complications. PMID- 19683143 TI - Simplified system of marking the cornea for a toric intraocular lens. AB - A 2-instrument set comprising a half-circle axis marker and a degree gauge that facilitates precise placement of ink marks on the cornea is described. Guiding the half-circle axis marker by the previously positioned degree gauge ensures placement of the marks on the astigmatic axis line that transects the center of the cornea and overlies the index marks on the toric intraocular lens. PMID- 19683145 TI - Functional visual outcomes of cataract surgery in patients with 20/20 or better preoperative visual acuity. AB - PURPOSE: To measure the visual outcomes after cataract surgery in patients with 20/20 or better preoperative visual acuity. SETTING: University-based cataract referral practice. METHODS: In this retrospective case review, 2 groups of patients with a corrected distance visual acuity (CDVA) of 20/20 or better in both eyes before cataract surgery were identified. Patients in the first group had a cataract in 1 eye and an intraocular lens (IOL) in the other eye. Patients in the second group had cataracts in both eyes. Each patient completed a Visual Function 14 (VF-14) questionnaire before and after cataract surgery. Changes in VF-14 scores were analyzed. RESULTS: The VF-14 scores improved significantly after cataract surgery in both groups. Patients with cataract in 1 eye and an IOL in the other eye before surgery (n = 28) had a mean improvement of 12.2 in the VF 14 score (P = .029). Patients with cataract in both eyes (n = 31) had a mean improvement of 14.5 in the VF-14 score (P<.0001). CONCLUSIONS: The functional vision of patients with 20/20 or better preoperative CDVA improved significantly after cataract surgery. This finding shows that arbitrary CDVA thresholds of worse than 20/20 cannot always be used to determine who will benefit from cataract surgery. PMID- 19683144 TI - Reading ability with pseudophakic monovision and with refractive multifocal intraocular lenses: comparative study. AB - PURPOSE: To the compare the reading ability after bilateral cataract surgery in patients who had pseudophakic monovision achieved by monofocal intraocular lens (IOL) implantation and patients who had refractive multifocal IOL implantation. SETTING: Department of Ophthalmology, Kitasato University Hospital, Kanagawa, Japan. METHODS: This study evaluated patients who had bilateral cataract surgery using the monovision method with monofocal IOL implantation to correct presbyopia (monovision group) or who had bilateral cataract surgery with refractive multifocal IOL implantation (multifocal group). In the monovision group, the dominant eye was corrected for distance vision and the nondominant eye for near vision. The maximum reading speed, critical character size, and reading acuity were measured binocularly without refractive correction using MNREAD-J acuity charts. RESULTS: The monovision group comprised 38 patients and the multifocal group, 22 patients. The mean maximum reading speed was 350.5 characters per minute (cpm) +/- 62.3 (SD) in the monovision group and 355.0 +/- 53.3 cpm in the multifocal group; the difference was not statistically significant. The mean critical character size was 0.24 +/- 0.12 logMAR and 0.40 +/- 0.16 logMAR, respectively (P<.05). The mean reading acuity was 0.05 +/- 0.12 logMAR and 0.19 +/- 0.11 logMAR, respectively (P<.01). The monovision group had better critical character size and reading acuity results. CONCLUSION: The monovision method group had better reading ability; however, careful patient selection is essential. PMID- 19683146 TI - Using Rasch analysis to revisit the validity of the Cataract TyPE Spec instrument for measuring cataract surgery outcomes. AB - PURPOSE: To assess the psychometric properties of the Cataract TyPE Specification (Spec) questionnaire using the Rasch model. SETTING: Flinders Medical Centre, Adelaide, Australia. METHODS: The 12-item Cataract TyPE Spec questionnaire was self-administered to patients drawn from the cataract surgery waiting list. The questionnaire and its 5 subscales were assessed for fit to the Rasch model. Response category performance, item-fit targeting, unidimensionality (using principal components analysis), and differential item functioning were assessed. A shortened version (11 items) was tested for criterion validity by determining correlation with a global rating of vision question. RESULTS: Two hundred ninety four patients responded to the questionnaire. The response categories for each question functioned as intended. Person-separation reliability was high (0.90). Deletion of 1 misfitting item (nighttime driving) improved overall model fit. The principal components analysis of the residuals demonstrated unidimensionality for the 11-item Cataract TyPE Spec and 2 subscales. However, items were targeted to a less able population. Only 2 subscales (near vision and glare) were valid. There was a good statistically significant correlation between the Likert-scored global rating of vision and the Rasch-scaled Cataract TyPE Spec score (r = -0.66, P<.0001), suggesting good criterion validity. CONCLUSIONS: With minor modifications, the Cataract TyPE Spec questionnaire and its glare and near-vision subscales were good measures of visual functioning in the cataract patient. Additional items to suit the more able, including patients having second-eye surgery, could improve the measurement properties. PMID- 19683147 TI - Combined cataract extraction and intravitreal bevacizumab in eyes with choroidal neovascularization resulting from age-related macular degeneration. AB - PURPOSE: To evaluate the safety and efficacy of phacoemulsification, intraocular lens (IOL) implantation, and a single intravitreal injection of bevacizumab in patients with coexisting visually significant cataract and subfoveal neovascularization due to age-related macular degeneration. SETTING: Department of Ophthalmology, University of Bari, Bari, Italy. METHODS: Eyes with predominantly classic subfoveal neovascularization and cataract had phacoemulsification, IOL implantation, and a 1.25 mg intravitreal injection of bevacizumab. One month after combined surgery, corrected distance visual acuity (CDVA), anterior chamber reaction, and intraocular pressure were evaluated and central foveal thickness was measured by optical coherence tomography. RESULTS: Twenty eyes of 20 patients were evaluated. One month postoperatively, the mean CDVA improved significantly, from 20/100 (range 20/160 to 20/80) at baseline to 20/63 (range 20/125 to 20/50) (P<.0001). The mean central foveal thickness decreased significantly, from 353.75 microm +/- 12.50 (SD) (range 334 to 375 microm) at baseline to 275.7 +/- 17.3 microm (range 255 to 323 microm) at 1 month (P<.0001). Intraocular pressure did not change significantly, and anterior chamber reaction was absent. No ocular or systemic adverse events were observed. CONCLUSION: Combined phacoemulsification, IOL implantation, and intravitreal bevacizumab was a safe and efficacious treatment in patients with visually significant cataract and active subfoveal neovascularization. PMID- 19683149 TI - Prospective evaluation of posterior capsule opacification in myopic eyes 4 years after implantation of a single-piece acrylic IOL. AB - PURPOSE: To determine whether axial myopia affects posterior capsule opacification (PCO) in eyes with high myopia (axial length [AL] > or = 26.00 mm) 4 years after implantation of an AcrySof single-piece intraocular lens (IOL). SETTING: Clinical practice, Ahmedabad, India. METHODS: In a prospective masked trial, consecutive eyes with high axial myopia (myopia group) and age-matched eyes with a normal AL (21.00 to 23.99 mm) (control group) had phacoemulsification and in-the-bag IOL implantation. Postoperatively, PCO was documented by standardized digital retroillumination images. The PCO area behind the IOL optic was scored (scale 0% to 100%) using the POCO software system. The presence of PCO within the central 3.0 mm of the optic was assessed with the Evaluation of Posterior Capsule Opacification software system. The incidence of neodymium:YAG (Nd:YAG) capsulotomy was determined. RESULTS: The median PCO in the myopia group and control group, respectively, was 3.2% and 5.6% at 1 month (P = .2), 2.2% and 1.6% at 12 months (P = .079), 7.0% and 2.3% at 24 months (P = .079), 3.7% and 3.6% at 36 months (P = .78), and 10.0% and 2.3% at 48 months (P = .61). By 4 years, PCO had encroached onto the central 3.0 mm of the optic in 38% in the myopia group and 20% in the control group (P = .04). One eye (2%) in each group had an Nd:YAG capsulotomy, at 3.2 years and 3.9 years, respectively. CONCLUSION: Axial myopia did not significantly increase the area or incidence of PCO at 4 years. PMID- 19683150 TI - Anterior segment optical coherence tomography findings of reverse pupillary block after scleral-fixated sutured posterior chamber intraocular lens implantation. AB - PURPOSE: To evaluate findings of reverse pupillary block after scleral-fixated sutured posterior chamber intraocular lens (PC IOL) implantation using anterior segment optical coherence tomography (AS-OCT) and report a possible association with postoperative complications. SETTING: Department of Ophthalmology, Kanazawa University Graduate School of Medical Science, Kanazawa, Japan. METHODS: Anterior segment OCT was used to evaluate eyes with reverse pupillary block (ie, deep anterior chamber, concave iris configuration, contact between pupil and IOL) after IOL explantation or cataract extraction combined with vitrectomy and scleral-fixated sutured PC IOL implantation. The PC IOL was implanted using an ab externo method in all cases. After 2 scleral flaps were created 180 degrees apart, a straight needle attached to a 10-0 polypropylene suture for IOL fixation was passed through the bed of half-thickness scleral flaps 2.0 mm posterior to the limbus in a direction parallel to the iris. RESULTS: Four eyes (3 patients) were evaluated. Anterior segment OCT showed a concave iris configuration with an increase in anterior chamber depth and anterior chamber angle in all cases. Two eyes had intermittent pupil capture. Laser iridotomy prevented recurrence of reverse pupillary block and pupil capture. Widening of the gap between the inner and outer flaps of the scleral tunnel during reverse pupillary block was seen on AS-OCT in 2 eyes; the widening was associated with wound leakage in 1 eye. CONCLUSION: Anterior segment OCT showed that reverse pupillary block can occur in eyes with scleral-fixated sutured PC IOLs and cause postoperative pupil capture and scleral tunnel insufficiency. PMID- 19683151 TI - Clinical outcomes and postoperative intraocular optical quality with a microincision aberration-free aspheric intraocular lens. AB - PURPOSE: To evaluate the visual outcomes and optical quality with an aspheric intraocular lens (IOL) with no aberration in microincision cataract surgery. SETTING: Vissum-Instituto de Oftalmologico de Alicante, Alicante, Spain. METHODS: This prospective cohort study included eyes that had cataract surgery with implantation of an Akreos Adapt Advanced Optics MI60 IOL. Surgery was performed using the Millennium phacoemulsification platform. The IOL was implanted through a sub-1.8 mm incision using the 1.8 Viscoglide cartridge. Visual and refractive outcomes were analyzed during a 12-month follow-up. In addition, postoperative ocular optical quality was evaluated using the Optical Quality Analysis System. Postoperative intraocular optical aberrations were calculated by subtracting corneal aberrations from total aberrations. RESULTS: The cohort comprised 25 eyes of 25 patients ranging in age from 52 to 83 years. The mean spherical equivalent was -0.47 diopters +/- 0.62 (SD) 3 months postoperatively (P = .72). The mean corrected distance visual acuity improved from 0.54 +/- 0.23 logMAR preoperatively to 0.08 +/- 0.16 logMAR 3 months postoperatively (P<.01). Optical quality analysis showed a mean spatial frequency at 50% of the modulation transfer function (MTF) of 2.85 +/- 0.55 cycles per degree (cpd) and a mean cutoff MTF frequency of 21.50 +/- 7.02 cpd. Postoperatively, the mean intraocular spherical aberration was 0.16 +/- 0.11 microm and the mean primary coma root mean square, 0.23 +/- 0.15 microm. CONCLUSION: Implantation of the aberration-free aspheric IOL was safe and effective and provided excellent visual and refractive outcomes with good optical performance. PMID- 19683148 TI - Clinical observations associated with proven and unproven cases in the ESCRS study of prophylaxis of postoperative endophthalmitis after cataract surgery. AB - PURPOSE: To describe cases of postoperative endophthalmitis in the European Society of Cataract & Refractive Surgeons (ESCRS) study of the prophylaxis of endophthalmitis, compare characteristics of unproven cases and cases proven by culture or polymerase chain reaction, and compare the characteristics with those in other reported series. SETTING: Twenty-four ophthalmology units in Austria, Belgium, Germany, Italy, Poland, Portugal, Spain, Turkey, and the United Kingdom. METHODS: Univariable and multivariable logistic regression models were used to analyze data for statistical association of signs and symptoms in cases with proven or unproven endophthalmitis. Specific data describing characteristics of the cases were compared between the 2 types of cases. RESULTS: Data from 29 endophthalmitis cases were analyzed. Swollen lids and pain were statistically associated with proven cases of endophthalmitis on univariable regression analysis. Multivariable analysis indicated that swollen lids and an opaque vitreous were associated with proven cases. Five cases of endophthalmitis occurred in the cefuroxime-treated groups. No case of streptococcal infection occurred in the cefuroxime-treated groups. However, cases of infection due to streptococci showed striking differences in visual acuity and were associated with earlier onset. Characteristics in the 29 cases parallel results in previous studies, such as the Endophthalmitis Vitrectomy Study, although the addition of a control group in the ESCRS study elicited additional findings. CONCLUSION: Swollen lids, pain, and an opaque vitreous were statistically associated with proven endophthalmitis cases in the ESCRS study. PMID- 19683152 TI - Astigmatic neutrality in biaxial microincision cataract surgery. AB - PURPOSE: To assess the astigmatic effect of biaxial microincision cataract surgery (MICS) with insertion of an UltraChoice 1.0 Rollable Thinlens intraocular lens (IOL) in a sufficiently powered controlled study. SETTING: Queen Elizabeth Hospital, South Australian Institute of Ophthalmology, University of Adelaide, Adelaide, Australia. METHODS: Consecutive patients having biaxial MICS were evaluated prospectively. Keratometry was performed preoperatively and postoperatively. Vector analysis of the alteration in the keratometric cylinder was compared with that in control eyes not having surgery but having keratometry over a similar time frame. RESULTS: There were 76 eyes in the surgical group and 74 in the control group. The right-hand incision used for IOL insertion had a mean external opening width of 2.00 mm and a mean internal width of 1.89 mm. The left-hand incision measured a mean of 1.49 mm and 1.46 mm, respectively. There was no statistically significant difference between the surgical group and the routine variability in keratometry in the untreated control group in surgically induced astigmatism (surgical group, 0.57 diopter [D] +/- 0.05 [SEM]; control group, 0.54 +/- 0.06 D; P =.660) or the degree of mean calculated flattening effect at the right-hand incision (0.01 +/- 0.06 D and -0.05 +/- 0.05 D, respectively; P = .405) or at the left-hand incision (-0.06 +/- 0.05 D and 0.03 +/- 0.06 D, respectively; P = .283). CONCLUSION: Biaxial MICS with insertion of IOL through a temporal incision of 2.0 mm offers prospects of astigmatic neutrality in cataract surgery. PMID- 19683153 TI - Comparison of surgically induced astigmatism after coaxial phacoemulsification through 1.8 mm microincision and bimanual phacoemulsification through 1.7 mm microincision. AB - PURPOSE: To compare surgically induced astigmatism (SIA) after coaxial 1.8 mm microincision cataract surgery (MICS) and bimanual 1.7 mm MICS calculated with 3 mathematical methods. SETTING: Department of Ophthalmology, Medical University of Lodz, Lodz, Poland. METHODS: Study comprised a nonrandomized prospective consecutive series of 58 eyes of 58 patients who had uneventful coaxial MICS with implantation of an Akreos MI60 foldable intraocular lens (IOL) using a 1.8 mm temporal clear corneal incision. Fifty eyes of 50 patients who had uneventful bimanual MICS through a 1.7 mm clear corneal incision for a sleeveless phaco tip and a 1.5 mm side port for an irrigating chopper with implantation of an Acri.Smart 48S foldable IOL served as a comparison group. All surgery was performed by 2 experienced surgeons. Surgically induced astigmatism was calculated using 3 methods. RESULTS: The patients were examined preoperatively and 2 weeks to 1 month postoperatively. No intraoperative or postoperative complications were seen in any patient. The corrected distance visual acuity improved significantly in both groups after surgery (P<.01); the visual outcomes were not significantly different (P>.05). In vector analysis, the mean SIA was 0.42 +/- 0.29 in the coaxial MICS group and 0.50 +/- 0.24 in the bimanual group; the difference was not statistically significant (P>.05). In vector decomposition, the mean SIA (C90) coaxial MICS group was 0.23 +/- 0.29 in the coaxial MICS group and 0.23 +/- 0.22 in the bimanual MICS group; the difference was not significant. Using the Naeser method, DeltaKP-90 was calculated, amounting to 0.05 +/- 0.44 in the coaxial MICS group and -0.04 +/- 0.42 in the bimanual MICS group; the difference was not significant. CONCLUSIONS: The amount of SIA induced by bimanual MICS and coaxial MICS phacoemulsification was very small. The bimanual MICS induced a slightly higher degree of SIA; however, according to all methods of SIA analysis, there was no significant difference in the mean SIA induced by both techniques. PMID- 19683155 TI - Intraocular lens power calculation and optimized constants for highly myopic eyes. AB - PURPOSE: To determine the accuracy of intraocular lens (IOL) power calculations in eyes with high myopia and to suggest adjusted constants for these cases. SETTING: Centre for Ophthalmology, Eberhard-Karls-University, Tuebingen, Germany. METHODS: Patients with high myopia having phacoemulsification with implantation of an AcrySof MA60MA IOL (power range +5.00 to -5.00 diopters [D]) were evaluated. Optical biometry (IOLMaster) and IOL calculations were performed before and after IOL implantation. Because of different optic principal planes of negative-diopter and positive-diopter IOLs, separate constants were calculated for these groups. RESULTS: Fifty eyes (32 patients) were evaluated. Thirty eyes (mean AL 31.15 mm +/- 1.69 [SD]) had implantation of a positive-diopter IOL (mean power +3.10 +/- 1.50 D) and 18 eyes (mean AL 33.20 +/- 2.25 mm), a negative diopter IOL (mean power -3.20 +/- 1.70 D). Postoperatively, the mean spherical equivalent was -1.42 +/- 1.33 D and -0.41 +/- 1.81 D, respectively. The difference in optimized constants between positive- and negative-diopter IOLs was significant for all formulas. Power calculation with the SRK II formula showed a wide range of deviation of postoperative refraction from target refraction. Calculation with the Haigis, SRK/T, Holladay 1, and Hoffer Q formulas showed a mean deviation of 0.00 D with an SD of 0.88, 0.92, 1.03, and 1.15, respectively. CONCLUSIONS: Results indicate that the SRK II formula cannot be recommended for IOL power calculation in highly myopic patients. With optimized constants, the SRK/T, Haigis, Hoffer Q, and Holladay 1 formulas produced small deviation of postoperative refraction from target refraction. PMID- 19683154 TI - Comparison of early corneal endothelial cell loss after coaxial phacoemulsification through 1.8 mm microincision and bimanual phacoemulsification through 1.7 mm microincision. AB - PURPOSE: To compare corneal endothelial cell loss after coaxial 1.8 mm microincision cataract surgery (MICS) and bimanual 1.7 mm MICS. SETTING: Department of Ophthalmology, Medical University of Lodz, Lodz, Poland. METHODS: The study comprised a nonrandomized prospective consecutive series of 51 eyes of 51 patients who had coaxial MICS with implantation of an MI60 foldable intraocular lens (IOL) using a 1.8 mm temporal clear corneal microincision. Fifty eyes of 50 patients who had uneventful bimanual MICS through a 1.7 mm temporal clear corneal incision for a sleeveless phaco tip and a side port for an irrigating chopper with a foldable Acri.Smart 48S foldable IOL implantation served as a reference group. Corneal endothelial cell density, intraoperative phaco power, effective phaco time, and preoperative and postoperative visual acuities were evaluated. The measurements were performed in a semiautomated masked manner. Statistical analysis was done using nonparametric tests (Wilcoxon signed rank test and Mann-Whitney U test). RESULTS: The patients were examined preoperatively and 2 weeks to 1 month postoperatively. The mean follow-up was 22.58 days +/- 5.08 (SD). Postoperatively, the mean corrected distance visual acuity (CDVA)was 0.95 +/- 13 in both groups. There was a significant decrease in endothelial cell density in both groups, 9.46% in Group 1 and 9.27% in Group 2. The between-group difference was not statistically significant (P>.05, Mann Whitney U test). CONCLUSIONS: The visual results were excellent in both groups. Both MICS techniques enabled preservation of corneal endothelial cells equally well and were similar in terms of minor surgical trauma and the influence of surgery on corneal endothelial cell density. Our results support the use of both MICS techniques for cataract surgery. PMID- 19683156 TI - Changes in vaulting and the effect on refraction after phakic posterior chamber intraocular lens implantation. AB - PURPOSE: To assess the changes in vaulting over time after implantable Collamer lens (ICL) implantation and the effect of vaulting on refraction. SETTING: Department of Ophthalmology, Kitasato University, Kanagawa, Japan. METHODS: This retrospective study evaluated eyes of consecutive patients having ICL implantation. The postoperative changes in vaulting between the ICL and the crystalline at 1, 3, and 6 months and 1 year were quantitatively assessed. The relationship between the vaulting and refractive outcomes at 1 year was also evaluated. RESULTS: Seventy-five eyes of 47 patients were evaluated. The mean vaulting was 0.61 mm +/- 0.26 (SD), 0.59 +/- 0.25 mm, 0.54 +/- 0.25 mm, and 0.53 +/- 0.24 mm at 1 month, 3 months, 6 months, and 1 year, respectively. The mean refractive error (difference between attempted and achieved manifest spherical equivalent refraction) was 0.01 +/- 0.42 diopter (D) 1 year postoperatively. There was no significant association between the amount of vaulting and the refractive error (r = 0.19, P = .11). CONCLUSIONS: Vaulting of the ICL over the crystalline lens decreased slightly with time, likely as a result of pupil movement, age-related increases in crystalline lens thickness, and the fixed position of the ICL haptics. The vaulting did not significantly affect refractive outcomes, suggesting that a precise effective lens position leads to higher predictability, largely as a result of the narrow fixated location of the ICL between the back surface of the iris and the ciliary sulcus. PMID- 19683157 TI - Intraocular lens power adjustment nomogram after laser in situ keratomileusis. AB - PURPOSE: To develop a nomogram for improving the accuracy of intraocular lens (IOL) power calculation at the time of cataract surgery based on the manifest refraction spherical equivalent (MRSE) change produced by laser in situ keratomileusis (LASIK). SETTING: University-based clinical practice, Jules Stein Eye Institute, Los Angeles, California, USA. METHODS: This was a retrospective review of the records of consecutive patients who had cataract surgery after LASIK. Spherical equivalent refractive data before and after LASIK and cataract surgery were recorded. The differences between the targeted postoperative refractive errors after cataract surgery and the achieved refractive errors were plotted against the LASIK-induced MRSE change, yielding postoperative refractive errors to target during IOL power calculation to achieve emmetropia. RESULTS: Thirty-two eyes of 23 patients were identified; 25 eyes had myopic LASIK and 7, hyperopic LASIK. Regression analysis yielded the following polynomial relationship: target postoperative refractive error (D) to achieve emmetropia during IOL power calculation = -0.018(MRSE Change)(2) + 0.192(MRSE Change) - 0.062. The outcomes in 97% of eyes fell within +/-1.00 D of the value predicted by this formula. CONCLUSIONS: A 2nd-order polynomial relationship was found between LASIK-induced MRSE change and the MRSE error after cataract surgery. From this equation, a simple adjustment nomogram was generated and put into a look-up table format. The formula and nomogram should improve IOL power calculation accuracy in post-LASIK eyes when the MRSE change is known. PMID- 19683158 TI - Regression of effect over long-term follow-up of conductive keratoplasty to correct mild to moderate hyperopia. AB - PURPOSE: To evaluate the long-term efficacy and stability of conductive keratoplasty (CK) for low to moderate hyperopia. SETTING: Department of Ophthalmology, Stanford University School of Medicine, Stanford, California, USA. METHODS: In this prospective nonrandomized noncontrolled clinical trial, performed as part of a U.S. Food and Drug Administration pivotal trial, patients with +0.75 to +3.00 diopters (D) of hyperopia and 0.75 D or less cylinder were initially treated with CK. The postoperative uncorrected and corrected Snellen visual acuities and refractions over time were evaluated. The main outcome measures were differences in refraction and uncorrected visual acuity postoperatively. RESULTS: Nine patients (16 eyes) of a subset of 14 consecutive patients (25 eyes) originally enrolled between 1999 and 2000 were available for long-term follow-up. Preoperatively, the mean manifest refraction spherical equivalent (MRSE) in the subset with long-term follow-up was +1.45 D. Postoperatively, the mean MRSE was +0.295 D. at 23 months, regressing to +1.00 D at 48 months and +1.394 D at the final follow-up (mean 73 months), indicating significant regression of effect. No eye lost more than 1 line of corrected Snellen visual acuity as a result of CK. The rate of regression was linear and calculated at +0.0184 D per month after 6 months postoperatively. Keratometric regression appeared to be similar to MRSE regression. CONCLUSIONS: Conductive keratoplasty led to significant regression of refractive and keratometric effects over extended follow-up. Patients should be counseled that this refractive procedure is not permanent. PMID- 19683159 TI - Keratoconus: classification scheme based on videokeratography and clinical signs. AB - PURPOSE: To determine in a longitudinal study whether there is correlation between videokeratography and clinical signs of keratoconus that might be useful to practicing clinicians. SETTING: Cornea-Genetic Eye Institute, Cedars-Sinai Medical Center, Los Angeles, California, USA. METHODS: Eyes grouped as keratoconus, early keratoconus, keratoconus suspect, or normal based on clinical signs and videokeratography were examined at baseline and followed for 1 to 8 years. Differences in quantitative videokeratography indices and the progression rate were evaluated. The quantitative indices were central keratometry (K), the inferior-superior (I-S) value, and the keratoconus percentage index (KISA). Discriminant analysis was used to estimate the classification rate using the indices. RESULTS: There were significant differences at baseline between the normal, keratoconus-suspect, and early keratoconus groups in all indices; the respective means were central K: 44.17 D, 45.13 D, and 45.97 D; I-S: 0.57, 1.20, and 4.44; log(KISA): 2.49, 2.94, and 5.71 (all P<.001 after adjusting for covariates). Over a median follow-up of 4.1 years, approximately 28% in the keratoconus-suspect group progressed to early keratoconus or keratoconus and 75% in the early keratoconus group progressed to keratoconus. Using all 3 indices and age, 86.9% in the normal group, 75.3% in the early keratoconus group, and 44.6% in the keratoconus-suspect group could be classified, yielding a total classification rate of 68.9%. CONCLUSIONS: Cross-sectional and longitudinal data showed significant differences between groups in the 3 indices. Use of this classification scheme might form a basis for detecting subclinical keratoconus. PMID- 19683160 TI - Clinical outcomes after implantation of a new intrastromal corneal ring with a 210-degree arc length. AB - PURPOSE: To report the clinical outcomes of implantation of a new Ferrara intrastromal corneal ring segment (ICRS) with a 210-degree arc length in eyes with keratoconus. SETTING: Private eye clinic, Belo Horizonte, Brazil METHODS: Statistical analysis included preoperative and postoperative uncorrected distance visual acuity (UDVA), corrected distance visual acuity (CDVA), refraction, and keratometry. RESULTS: Eighty eyes (76 patients) were evaluated. The UDVA improved from 20/350 preoperatively to 20/136 postoperatively and the CDVA, from 20/125 to 20/50; the differences were statistically significant (P = .001 and P = .0001, respectively). Corneal tomography (Pentacam) showed corneal flattening in all eyes. The mean preoperative spherical equivalent decreased from -5.22 diopters (D) preoperatively to -2.26 D postoperatively (P = .050); the mean keratometry (K) 1 value, from 51.49 D to 47.40 D (P = .00014); and the mean K2 value, from 54.33 D to 49.14 D (P = .00022). Two patients required penetrating keratoplasty despite ICRS implantation. CONCLUSIONS: A new ICRS with 210 degrees of arc was effective in treating keratoconus. It improved visual acuity and reduced corneal steepening in selected patients. PMID- 19683161 TI - Kill rates of preserved and preservative-free topical 8-methoxy fluoroquinolones against various strains of Staphylococcus. AB - PURPOSE: To assess the kill rates of preserved and preservative-free 8-methoxy fluoroquinolones and benzalkonium chloride (BAC) against isolates of Staphylococcus aureus and coagulase-negative Staphylococcus (CNS) species. SETTING: Bascom Palmer Eye Institute, Miami, Florida, USA. METHODS: Ocular and standard isolates of S aureus and CNS were inoculated with saline, 0.005% BAC, gatifloxacin 0.3% containing 0.005% BAC (Zymar), or BAC-free moxifloxacin 0.5% (Vigamox) at 37 degrees C. Bacterial viability was assessed after 15 minutes, 30 minutes, and 60 minutes. RESULTS: Thirteen isolates of S. aureus and 5 isolates of CNS were used. The mean initial load of bacterial inoculum was 5.45 log colony forming units (CFU)/mL +/- 0.37 (SD). Saline did not affect the density of staphylococci; BAC significantly decreased the staphylococci count to a mean of 3.80 +/- 0.32 log CFU/mL at 60 minutes (P<.05). Gatifloxacin 0.3% reduced the number of staphylococci significantly more than moxifloxacin 0.5% at 15 minutes (mean 0.47 +/- 1.12 log CFU/mL versus 4.55 +/- 0.60 log CFU/mL), 30 minutes (mean 0.07 +/- 0.31 log CFU/mL versus 3.82 +/- 1.07 log CFU/mL), and 60 minutes (mean 0.00 +/- 0.00 log CFU/mL versus 2.75 +/- 1.29 log CFU/mL) (P<.005). Gatifloxacin 0.3% completely eradicated most S. aureus (10/13) and CNS (3/5) isolates at 15 minutes; moxifloxacin 0.5% did not achieve complete kill in any S. aureus isolate (13/13) or in most CNS isolates (4/5) at 60 minutes. CONCLUSION: The commercial formulation of gatifloxacin 0.3% containing BAC 0.005% completely eradicated staphylococcal isolates more rapidly than the BAC-free formulation of moxifloxacin 0.5% or BAC 0.005% alone. PMID- 19683163 TI - Allopatric femtosecond laser gas-bubble formation in a closed system. AB - PURPOSE: To test a possible mechanism for allopatric gas-bubble formation in femtosecond laser-assisted surgery using an experimental closed system. SETTING: Bascom Palmer Eye Institute, University of Miami, Miami, Florida, USA. METHODS: Plastic bottles were filled with balanced salt solution (BSS). Femtosecond laser (IntraLase) flap-mode cuts were made on the walls of the BSS bottles. After femtosecond laser application, the BSS bottles were viewed under the excimer laser microscope and the results photographed. Next, the BSS bottles and laser cuts were imaged with optical coherence tomography. RESULTS: In-bottle gas bubbles were consistently produced in the closed system. No cut lines crossed the inner wall of the BSS bottles. CONCLUSION: Results show that in situ gas-bubble formation as a mechanism for gas-bubble formation is a possible alternative to gas traveling through the anterior segment and into the intraflap laser space. PMID- 19683162 TI - Efficacy of ophthalmic nonsteroidal antiinflammatory drugs in suppressing anterior capsule contraction and secondary posterior capsule opacification. AB - PURPOSE: To evaluate the efficacy of ophthalmic nonsteroidal and steroidal antiinflammatory drugs in preventing anterior capsule contraction and secondary posterior capsule opacification (PCO) using an experimental cataract model. SETTING: Department of Ophthalmology, Dokkyo Medical University, Tochigi, Japan. METHODS: Eight-week-old albino rabbits weighing about 2 kg each had phacoemulsification and intraocular lens implantation. After surgery, the rabbits were divided into 3 treatment groups: diclofenac sodium ophthalmic solution, bromfenac sodium ophthalmic solution, and betamethasone ophthalmic solution. In each group, the ophthalmic solution was applied to the left eye of each rabbit twice daily; the right eye served as an untreated control. To evaluate anterior capsule contraction, the percentage of incised anterior capsule opening area was calculated on diaphanoscopic images obtained with an EAS-1000 anterior segment analyzer. For evaluation of PCO, a tissue section was stained with hematoxylin eosin and observed under a light microscope. The PCO was quantified on the basis of the thickness of the lens epithelial cell layer on the central subcapsular area and compared among groups. RESULTS: Fifteen albino rabbits were used in the study. Treatment with diclofenac sodium and bromfenac sodium ophthalmic solution prevented progression of anterior capsule contraction and PCO. Treatment with bromfenac ophthalmic solution did not prevent either complication. CONCLUSION: Postoperative treatment with ophthalmic nonsteroidal antiinflammatory drug solutions prevented anterior capsule contraction and PCO in rabbit eyes. PMID- 19683164 TI - Incorporating corneal pachymetry into the management of glaucoma. AB - Intraocular pressure (IOP) results from a dynamic balance between aqueous humor formation and outflow. The simplest technique to measure IOP is indentation tonometry. Another technique is applanation. These methods are related to the elasticity of the eye, which mainly depends on its thickness and hysteresis. For several decades, Goldmann applanation tonometry has been the most accepted method of measuring IOP; the Goldmann tonometer is still used in all important trials. The relationship between IOP values and central corneal thickness (CCT) is well known; Goldmann stated that this relationship only holds for an average corneal thickness of 520 microm measured by optical pachymetry. The Ocular Hypertension Treatment Study (OHTS) showed that CCT is an important risk factor for a change from ocular hypertension to primary open-angle glaucoma. In a multivariate model that included IOP, CCT was the most powerful component of the predictive model. In the Early Manifest Glaucoma Trial (EMGT) with an 11-year follow-up, CCT was a significant predictive factor for glaucoma progression in patients with higher baseline IOP but not in those with lower baseline IOP. Clinical trials such as the OHTS and EMGT cannot prove that CCT is linked to a risk for glaucoma on a biological level. Thus, in eyes with glaucoma, IOP must be treated because it has a significant influence on progression of glaucoma, regardless of the baseline IOP and CCT. PMID- 19683165 TI - Resident laser in situ keratomileusis surgical training in United States residency programs. AB - To determine current trends in resident laser in situ keratomileusis (LASIK) training in the United States, a comprehensive survey was sent to the residency coordinator, chairman, or refractive surgery director of all 113 ophthalmology training programs in the U.S. accredited by the Accreditation Council for Graduate Medical Education. The response rate was 64%. Of the respondents, 54% were from programs in which residents performed LASIK surgery, typically as third year residents; residents in these programs performed a mean of 4 LASIK surgeries (range 1 to 10 surgeries) during residency. The Visx Star was the most commonly used excimer laser platform and the Hansatome the most commonly used microkeratome. Most programs had 1 attending physician teaching the residents refractive surgery, which was typically performed at the main residency facility. Most programs had significant discounted pricing (>50%) for resident-performed LASIK. PMID- 19683167 TI - Bilateral intraocular hemorrhage from vascularization of cataract wounds. AB - We report a case of bilateral intraocular hemorrhage from vascularization of cataract wounds. The patient experienced decreased vision following an episode of vomiting more than 2.5 years after phacoemulsification through a scleral tunnel incision in the right eye and combined trabeculectomy and extracapsular cataract extraction in the left eye. Gonioscopy demonstrated abnormal vessels in the region of the cataract wound superiorly and small hyphemas inferiorly in both eyes. The hemorrhages and elevated intraocular pressure normalized over weeks. The left eye had a recurrent hemorrhage 5 months later, which was successfully treated with argon laser goniophotocoagulation. PMID- 19683166 TI - Endophthalmitis due to exposure of anterior chamber intraocular lens haptic tip. AB - After bilateral implantation of an angle-supported anterior chamber intraocular lens (AC IOL) in an allergic patient, the haptic tips were exposed through the limbus in the left eye. Endophthalmitis developed in the eye and was treated by pars plana vitrectomy, IOL extraction, and intravitreal antibiotic injection. Haptic repositioning was performed in the right eye. In this report, we assess the possibility that exposure of the angle-supported AC IOL haptics caused the endophthalmitis. We hypothesized that vigorous rubbing of the eye of the atopic patient might cause limbal erosion that could lead to exposure of the angle supported AC IOL haptic through the limbus. PMID- 19683168 TI - High-resolution optical coherence tomography visualization of LASIK flap displacement. AB - Uneventful myopic laser in situ keratomileusis (LASIK) was performed in both eyes of a 33-year-old woman. Two weeks after LASIK, examination of the left eye revealed flap striae radiating inferonasally from the superior hinge. The flap was relifted and repositioned to remove the irregular astigmatism and reduce the striae. Before the flap was relifted, Fourier-domain optical coherence tomography (FD-OCT) showed the gap at the temporal flap, which had not been detected by biomicroscopy. After the flap was relifted, FD-OCT documented that the gap was closed. High-resolution FD-OCT was helpful in the visualization and management of flap displacement. PMID- 19683169 TI - Endophthalmitis following cataract surgery: the sucking corneal wound. PMID- 19683171 TI - Deliberate use of unequal adds with aspheric ReSTOR intraocular lens. PMID- 19683170 TI - Serratia marcescens keratitis after photorefractive keratectomy. PMID- 19683172 TI - Consultation section. A dense opaque bubble layer (OBL) appeared to interfere with the laser dissection. PMID- 19683176 TI - Intraocular lens power after refractive surgery: Haigis-L formula. PMID- 19683179 TI - 10-point analog scale not equivalent to a 10-point questionnaire. PMID- 19683180 TI - Power calculation after laser refractive surgery. PMID- 19683181 TI - Extrapolating anti-vascular endothelial growth factor therapy into pediatric ophthalmology: promise and concern. PMID- 19683182 TI - What can we learn from computational model studies of the eye? PMID- 19683183 TI - Morning glory disk anomaly--more than meets the eye. PMID- 19683184 TI - The relationship between preoperative alignment stability and postoperative motor outcomes in children with esotropia. AB - PURPOSE: To examine the effect of preoperative alignment stability on postoperative motor outcomes in children who underwent surgery for esotropia. METHODS: One hundred sixty-seven subjects (68 with infantile esotropia and 99 with acquired esotropia) aged less than 6 years had surgery after completing 18 weeks of follow-up as part of an observational study. Preoperative alignment was classified as stable, uncertain, or unstable, based on measurements taken at baseline and every 6 weeks for 18 weeks. Distance alignment measured by prism and alternate cover test was compared among stability classification groups at 6 weeks and 6 months after surgery. RESULTS: Among subjects with infantile esotropia, median 6-week postoperative deviation was 2(Delta), 6(Delta), and 2(Delta) for subjects with stable, uncertain, and unstable preoperative alignment, respectively (p = 0.73 for stable vs unstable). Median 6-month postoperative deviation was 1(Delta), 9(Delta), and 1(Delta) for stable, uncertain, and unstable, respectively (p = 1.00 for stable vs unstable). Among subjects with acquired esotropia, median 6-week postoperative deviation was 6(Delta), 4(Delta), and 4(Delta) for subjects with stable, uncertain, and unstable preoperative alignment, respectively (p = 0.69 for stable vs unstable). Median 6-month postoperative deviation was 8(Delta), 4(Delta), and 6(Delta) for stable, uncertain, and unstable, respectively (p = 0.22 for stable vs unstable). CONCLUSIONS: Postoperative alignment at 6 weeks and 6 months appears similar in children with stable versus unstable preoperative esotropia. Nevertheless, our finding should be interpreted with caution due to small sample size. PMID- 19683185 TI - Results of bilateral medial rectus muscle recession in unilateral esotropic Duane syndrome. AB - PURPOSE: To report the results of bilateral medial rectus muscle recession in improvement of the ocular alignment and motility of patients with unilateral esotropic Duane syndrome. METHODS: All medical files of patients with Duane syndrome seen by the first author between 1997 and 2006 were reviewed. Pre- and postsurgical deviation, angle of abnormal head position, severity of limitation in abduction, severity of globe retraction, and upshoots and downshoots were compared. RESULTS: Twenty-five patients with unilateral esotropic Duane syndrome (type 1) underwent recession of both medial rectus muscles. Mean age of patients was 9.7 years: male-to-female ratio was 0.67 (10:15). The left eye was involved in 23 of the patients. Mean esotropia decreased from 24.3(Delta) (range, 12(Delta)-50(Delta)) to 1.3(Delta) (range, 0(Delta)-10(Delta)). Mean abnormal head position decreased from 21.4 degrees (range, 15 degrees-35 degrees) to 1 degrees (range, 0 degree-5 degrees). Esotropia and abnormal head position disappeared in 80% of the patients and improved in the remaining. None of the patients developed exotropia. Mean limitation in abduction decreased from -3.8 to -3.3. Globe retraction was eliminated in 14 of the patients and improved in the others. Mild upshoots and downshoots, which were observed in 5 patients, disappeared in 2 and improved in 3 patients. CONCLUSIONS: Bilateral recession of medial rectus muscles has resulted in improvement of deviation, abnormal head position, and globe retraction in patients with unilateral esotropic Duane syndrome. PMID- 19683186 TI - Performance of strabismic subjects using a validated surgical training module: a pilot study. AB - PURPOSE: To compare the performance of patients with strabismus to that of age matched controls in a validated surgical training module. METHODS: A prospective experimental study was conducted of 14 adult patients with strabismus since childhood and absent stereopsis and of 14 age-matched controls with normal stereopsis. Each participant received instruction in the task of peg transfer on a validated surgical training device and then completed 10 consecutive timed trials. The means of the best 5 scores were compared using the 2-sample Wilcoxon rank-sum test. RESULTS: The average age of cases was 34.8 years (range, 15-51 years) compared with 37.8 years (range, 14-56 years) for controls. The scores for the strabismic patients ranged from 50.8 to 151.4 seconds, with a mean of 82.5 +/ 26.7 seconds. Controls ranged from 43.2 to 129 seconds, with a mean of 64.7 +/- 23.9 seconds. The Wilcoxon rank-sum test showed significantly better performance among controls (p = 0.022). CONCLUSIONS: Patients with strabismus performed more poorly than did age-matched controls in this model of hand-eye coordination. However, there was significant overlap between groups and several patients with strabismus performed better than the mean of the control group. Further investigation is required to elucidate the impact of strabismus on surgical performance. PMID- 19683187 TI - Cyanoacrylate adhesive use in primary operation and reoperation in rabbit eye muscle surgery. AB - PURPOSE: To compare the performance of 2-octyl-cyanoacrylate to 6-0 polyglactin 910 suture in rabbit superior rectus muscle surgery after operation and reoperation procedures. METHODS: A prospective noninferiority trial was conducted in rabbits. Bilateral superior rectus muscle recessions were performed using cyanoacrylate in one eye and polyglactin suture in the other. At 5 weeks, reoperations to advance the superior rectus muscle were performed on 20 rabbits. Slippage, tensile strength, ease of reoperation, operative time, and inflammatory reaction were recorded. For the primary outcomes, the predetermined margin of noninferiority was 1 mm for slippage and 100 g for tensile strength. RESULTS: In both groups, the proportion of slippage > or =1 mm from the recession site was 1.9%. For the reoperation, it was 36.8% and 15.7% in the suture and cyanoacrylate groups, respectively, and the mean slippage was 0.60 mm and 0.42 mm. Mean tensile strength was 842.8 g for suture and 777.2 g for cyanoacrylate after the first operation and 877.73 g and 844.87 g after the reoperation. There was no difference between groups for surgical difficulty or inflammatory index. For the first operation, surgery using cyanoacrylate was on average 3.85 min faster than suture. CONCLUSIONS: Cyanoacrylate can achieve an adequate muscle-sclera bond in the immediate period after surgery to avoid major slippage and does not affect the long-term process of wound healing for both recession and advancement procedures. It is well tolerated and does not add technical difficulty even if used for reoperations. Because it eliminates the risk of globe perforation, cyanoacrylate may be a good alternative to sutures in strabismus surgery. PMID- 19683188 TI - Retinopathy of prematurity in extremely premature infants. AB - INTRODUCTION: The incidence and severity of retinopathy of prematurity (ROP) in extremely premature infants have not been reported since publication of the Early Treatment of ROP study results. The survival rate of these infants continues to increase. We sought to determine the characteristics of ROP in a group of surviving infants <25 weeks estimated gestational age (EGA) at birth compared to a group 25 to 27 weeks EGA at birth. METHODS: Retrospective review of infants born prior to 27 weeks EGA between January 2003 and July 2007 at a level-3 nursery at a regional academic medical center. RESULTS: A total of 231 medical records were reviewed and found to have analyzable data. Of 79 infants <25 weeks EGA, 69 (87%) developed ROP, compared to 95 of 152 (62%) infants 25 to 27 weeks EGA. Type 1 ROP developed in 23% of infants in the <25 weeks EGA group, compared to 9% of infants in the 25 to 27 weeks EGA group. There was no difference in mean postmenstrual age when type 1 ROP was diagnosed. Lower birth weight predicted increased risk of type 1 ROP in the 25 to 27 weeks EGA group, but not in the <25 weeks EGA group. CONCLUSIONS: Extremely premature infants are more likely to develop ROP and type 1 ROP, but the incidence may be lower than previously reported. Birth weight may not influence the incidence of type 1 ROP in this group of infants. Type 1 ROP does not develop at an earlier postmenstrual age in the extremely premature infant. PMID- 19683189 TI - Port-wine vascular malformations and glaucoma risk in Sturge-Weber syndrome. AB - PURPOSE: Treatment of the capillary vascular malformation (port-wine stain) in Sturge-Weber syndrome with the use of a laser is helpful cosmetically. However, concerns have been raised that laser obliteration of port-wine stains may result in ocular hypertension. The aim of this study was to review clinical features and management of ocular complications of SWS and assess the effects of dermatological laser treatment on the incidence of glaucoma or ocular hypertension. METHODS: This retrospective cohort study was conducted in an institutional setting. All patients had involvement of the face. Patients who underwent skin laser to the port-wine vascular malformation were analyzed further. Ocular involvement, glaucoma, and skin laser treatment and the relationship to ocular hypertension/glaucoma were observed. RESULTS: Forty-one Sturge-Weber syndrome patients with port-wine vascular malformation were analyzed. Glaucoma was observed in 24 patients (58.5%) at mean age of 2.9 years (range, 0.0-16.5). Of these, 18 (75.0%) were treated with medical therapy, and 10 (41.7%) required trabeculectomy, with 2 of these requiring Seton implant. Of the 41 patients, 28 (68.3%) underwent laser to face/forehead. Mean age of laser commencement was 5 years (range, 0.4-16.5). Thirteen did not undergo laser treatment. Fourteen of the 28 and 10 of the 13 developed ocular hypertension/glaucoma. CONCLUSIONS: This retrospective review did not find evidence to suggest that laser treatment of port-wine vascular malformations causes glaucoma or that it can worsen a preexisting ocular hypertension or glaucoma. Statistical analysis was inconclusive. PMID- 19683190 TI - Cyclophotocoagulation versus sequential tube shunt as a secondary intervention following primary tube shunt failure in pediatric glaucoma. AB - PURPOSE: To examine the efficacy of a sequential tube shunt versus transscleral diode cyclophotocoagulation following failure of an initial tube shunt on maximal medical therapy in treatment of refractory childhood glaucoma. METHODS: A nonrandomized retrospective chart review was conducted of 17 eyes of 14 pediatric patients (less than 18 years old) with refractory glaucoma treated with either sequential tube shunt (Group A) or diode cyclophotocoagulation (Group B) following initial failed tube shunt. Success was defined as an intraocular pressure < or =22 mm Hg on medical therapy, no visually devastating complications, and no further glaucoma surgery performed or recommended. RESULTS: Of the 17 eyes, 8 had a sequential tube shunt and 9 underwent diode cyclophotocoagulation as a secondary procedure. Kaplan-Meier analysis demonstrated a successful outcome of 75% and 62.5% at 12 months and 24 months, respectively, for Group A, and 66.7% at both 12 months and 24 months for Group B (p = 0.48). Corneal decompensation or graft failure was noted in 3/8 eyes (38%) in Group A. Cataract surgery was performed in 2/5 phakic eyes (40%) in Group B. One eye in each group progressed to no light perception. CONCLUSIONS: Diode cyclophotocoagulation and sequential tube shunt following primary tube shunt failure in childhood glaucoma showed similar efficacy and complication rates. However, the small sample size of this study warrants further evaluation of these 2 procedures following failure of a tube shunt device in pediatric glaucoma. PMID- 19683191 TI - Betaxolol hydrochloride ophthalmic suspension 0.25% and timolol gel-forming solution 0.25% and 0.5% in pediatric glaucoma: a randomized clinical trial. AB - PURPOSE: To describe the safety profile and clinical response on elevated intraocular pressure (IOP) of betaxolol hydrochloride ophthalmic suspension 0.25% (betaxolol) and timolol maleate ophthalmic gel-forming solution (TGFS) (0.25% and 0.5%), in subjects under 6 years of age. METHODS: Subjects were randomized to betaxolol 0.25% (twice daily) or TGFS (daily) (0.25% or 0.5%) in this double masked study. IOPs were obtained at the same time of day (9 AM) at 2 baseline visits and weeks 2, 6, and 12. Mean change from baseline in IOP was the primary efficacy parameter. RESULTS: One hundred five subjects were randomized (34 to betaxolol, 35 to TGFS 0.25%, 36 to TGFS 0.5%). Betaxolol, TGFS 0.25%, and TGFS 0.5% produced statistically significant mean reductions in IOP; mean reductions after 12 weeks of treatment were 2.3, 2.9, and 3.7 mm Hg, respectively. In subjects who were not being treated with topical IOP-lowering medication at baseline, mean IOP reductions after 12 weeks of treatment were 3.1, 4.8, and 3.8 mm Hg, respectively. In patients discontinuing 1 or more topical IOP-lowering medications at baseline, mean IOP reductions at Week 12 were 1.8, 1.8, and 3.7 mm Hg, respectively. Responder rates (> or =15% reduction from baseline) for betaxolol, TGFS 0.25%, and TGFS 0.5% were 38.2, 45.7, and 47.2%, respectively. Adverse events were predominantly nonserious and did not interrupt patient continuation in the study. CONCLUSIONS: Betaxolol ophthalmic suspension 0.25%, TGFS 0.25%, and TGFS 0.5% were well tolerated. Despite low responder rates, all 3 treatments produced statistically significant mean reductions in IOP in pediatric glaucoma subjects. PMID- 19683192 TI - Ocular findings in childhood-onset Behcet disease. AB - PURPOSE: To investigate the demographic characteristics and ocular features of patients with childhood-onset Behcet disease. PATIENTS AND METHODS: Patients for this retrospective study were selected using the data obtained from medical records of 34 patients with childhood-onset Behcet disease being followed up in the Uveitis Department of Ankara Ulucanlar Eye Education and Research Hospital between January 2003 and May 2008. RESULTS: Of 34 patients, 24 were boys (70.5%) and 10 were girls (29.4%). The average age of patients was 14.5 +/- 1.9 years (range, 10-16 years). Mean follow-up period was 4.5 +/- 2 years (range, 2-10 years). During the follow-up, panuveitis, posterior uveitis, and anterior uveitis were diagnosed in 18 (52.9%), 11 (32.3%), and 5 (14.7%) patients, respectively. Ocular findings included cataract in 20 patients (58.8%), posterior synechiae in 8 (23.5%), posterior capsular opacification secondary to cataract surgery in 8 (23.5%), vitreous condensation due to previous episodes of vitritis in 17 (50%), optic atrophy in 10 (29.4%), cystoid macular edema in 5 patients (14.7%), narrowed and occluded retinal vessels due to retinal periphlebitis and branched retinal vein occlusion in 2 (5.8%), neovascularization of the disk in 1 (2.9%), and phthisis bulbi in 1 patient (2.9%). CONCLUSIONS: Based on the results that were obtained in this series, panuveitis was the most common type of uveitis in cases with childhood-onset Behcet disease. Cataract was the most common anterior segment complication. Optic atrophy was the most common posterior segment complication. Male predominance was determined in this study. PMID- 19683193 TI - Ocular manifestations in chronic granulomatous disease in Saudi Arabia. AB - INTRODUCTION: Chronic granulomatous disease (CGD) is a primary immunodeficiency disease caused by a genetic defect in the NADPH oxidase complex of phagocytic cells. Recent reports indicate that chorioretinal lesions are more common than previously suspected. In this study, ocular findings of CGD patients are described with particular emphasis on chorioretinal lesions as a potentially serious ocular complication of CGD. METHODS: Medical records of CGD patients attending an immunodeficiency clinic at a tertiary care center from January 2004 to December 2006 were reviewed. Patients underwent full ophthalmologic examination. Patients with chorioretinal lesions were investigated for various causes of chorioretinitis. Molecular studies for common CGD-causing genes were performed in patients with chorioretinal lesions. RESULTS: This cohort included 32 CGD patients: 14 (44%) had abnormal eye findings, 11 (34%) had anterior segment disease, and 4 (12.5%) had chorioretinal lesions. Posterior segment findings consisted of uniformly similar hypopigmented atrophic punched-out chorioretinal scars around the arcades and mid-equator sparing of the macula. One patient had exudative hemorrhagic total retinal detachment in the right eye. Two siblings with chorioretinal lesions had mutation in CYBB, an X-linked gene. Another patient carried a missense mutation in NCF2, causing autosomal-recessive disease. CONCLUSIONS: While ocular manifestation is common in CGD, chorioretinal lesions seem less frequent. However, they present potential risk of visual loss; it is recommended that patients undergo regular ophthalmologic examinations. This report provides further evidence that chorioretinal lesions occur not only in X linked, but they can also occur in the autosomal-recessive form of CGD. PMID- 19683194 TI - Morning glory disk anomaly: a computerized analysis of contractile movements with implications for pathogenesis. AB - Excavated optic disk anomalies, such as morning glory disk anomaly, peripapillary staphyloma, and optic disk coloboma, are rare. Contractile movement of a given lesion is even rarer. Only 4 patients with morning glory disk anomalies accompanied by contractile movement have been reported. We offer a computerized analysis of the contraction, observable on a video recording of a patient with this disorder, with an explanation of the possible pathogenesis of the disorder. PMID- 19683196 TI - Repair of cicatricial ectropion in a harlequin baby. AB - The ichthyoses are a collection of scaling skin diseases or keratinizing skin disorders giving the appearance of "fish skin," of which harlequin ichthyosis is the most severe form. It is characterized by profound thickening of the keratin skin layer, armorlike scales that cover the body, and contraction abnormalities of the eyes, ears, and mouth. We report a case of a 6-week-old boy with harlequin ichthyosis and severe bilateral upper and lower eyelid cicatricial ectropion who underwent surgical repair with full-thickness postauricular skin autografts. To our knowledge, this is the youngest reported case and the only case of harlequin ichthyosis in which postauricular skin grafts were used. PMID- 19683195 TI - Unilateral eyelid angiofibroma with complete blepharoptosis as the presenting sign of tuberous sclerosis. AB - Tuberous sclerosis is a multisystem autosomal-dominant disease characterized by hamartomatous growths in the brain, skin, kidneys, eyes, and heart, but it may affect almost any organ. Retinal hamartomas are 1 of the major diagnostic criteria for tuberous sclerosis and occur in approximately 50% of patients. Nonretinal findings include angiofibromas of the eyelid, strabismus, and pseudo colobomas of the lens and iris. We report a case of a newborn with congenital eyelid angiofibroma mimicking complete congenital blepharoptosis that was revealed by central nervous system imaging to be part of the tuberous sclerosis complex. PMID- 19683197 TI - Congenital trigemino-abducens synkinesis in a neonate. AB - Congenital ocular synkinesis syndromes involve aberrant innervation of extraocular and eyelid muscles in a variety of patterns. A rare iteration is trigemino-abducens synkinesis, with only three published cases to date. Here the authors report (with video documentation) the earliest documented age of trigemino-abducens synkinesis and congenital ocular synkinesis in general. A 13 week-old (40-week postmenstrual age) girl presented with rhythmic abduction of the left eye that coordinated with sucking, likely resulting from abnormal embryologic development, causing activation of the lateral rectus by motor fibers of the mandibular branch of the trigeminal nerve. PMID- 19683198 TI - Horizontal mattress technique for superior oblique suture spacer. AB - Superior oblique tendon overaction and Brown syndrome have been managed surgically by means of tenotomy/tenectomy, use of a silicon expander, and elongation with autologous fascia lata. These solutions are problematic with respect to corrective precision, injury and complications, or surgical difficulty. We present a simple "mattress" suture spacer technique that allows a precise and secure intraoperative adjustment. PMID- 19683199 TI - Exotropia caused by pit viper snakebite. AB - Pit viper snake venom exerts a mildly neurotoxic effect that may rarely lead to neurotoxic complications, such as paralytic strabismus. Extraocular muscles are known to be particularly susceptible to this complication. In previously reported cases, the medial rectus muscle has been the most frequently involved. We report the first case of a pit viper snakebite resulting in comitant exotropia without obvious paralytic features. PMID- 19683201 TI - Primary treatment of nasolacrimal duct obstruction. PMID- 19683203 TI - Long-term follow-up of congenital esotropia in a population-based study. PMID- 19683204 TI - Twenty years of experience with PDD and PDT in Poland--review. AB - This article is a review of laboratory and clinical research undertaken in Poland in PDD and PDT over the past 20 years. These are divided into two parallel research areas. The first is based on clinical trials where new modalities of photosensitizer synthesis, molecular mechanisms of PDT and other aspects are investigated. The second is concerned with clinical aspects of PDD and PDT in both pre-neoplastic and malignant disease. In Poland there were 2 National Congresses in 2006 and 2008 with 100 and 400 participants respectively. One of the oldest centers of Photodynamic Diagnostics and Therapy is located in Bytom. For about 10 years it has led clinical research in Poland with PDD and PDT in such medical disciplines as dermatology, gastroenterology, laryngology, pulmonology, gynecology, and orthopedics. PMID- 19683205 TI - A brief history of photodynamic therapy in Wroclaw. AB - A brief history of photodynamic therapy in Wroclaw was presented in this paper. PMID- 19683206 TI - Cholangiocarcinoma: improving biliary drainage with PDT. PMID- 19683207 TI - Cholangiocarcinoma: an emerging indication for photodynamic therapy. AB - Cholangiocarcinoma (CC) is emerging as an important treatment indication for photodynamic therapy. CCs are generally unresectable locally invasive tumors that occlude the biliary tree leading to fatal cholangitis and liver failure. Biliary decompression via stenting offers symptomatic relief but does not control tumor growth. Founded on an initial case study followed by ever more sophisticated clinical research, including randomized trials, photodynamic therapy has garnered enough momentum to be considered as part of the standard of care for these patients. Further, preliminary clinical data show the potential for benefit of the use of PDT in a neoadjuvant and adjuvant fashion to the minority of patients currently considered resectable or of border line resectability. PDT also impacts interleukin-6 levels and may form the basis for a targeted therapy approach to this disease. We review the clinical rationale, current studies and potential future directions of PDT for patients with CC. PMID- 19683208 TI - Comment for the review of Dr Allison. PMID- 19683209 TI - Chlorin e6-based photosensitizers for photodynamic therapy and photodiagnosis. PMID- 19683210 TI - Pharmacokinetics and biodistribution of Photolon (Fotolon) in intact and tumor bearing rats. AB - BACKGROUND: This paper provides the results of the non-clinical evaluation of biodistribution of the PS Photolon in inner organs and tissues of intact and tumor-bearing rats with xenograft tumors of different morphologic types. METHODS: The accumulation studies were performed in rats by means of intravital laser fluorimetry in situ and spectrophotometric determination ex vivo. RESULTS: The biodistribution pattern of Photolon does not depend on tumor carriage as well as on morphologic type of the xenograft tumor. We have also showed that Photolon easily crosses an intact blood-brain barrier and accumulates in tissues of central nervous system. The relative bioavailability of brain tissues for Photolon was estimated as 82%, T(max)-30 min, mean residual time (MRT)-1.6h. CONCLUSIONS: In general, results of the experimental study of biodistribution of Photolon in inner organs and tissues of rats, performed as in real time (by means of intravital laser fluorimetry in situ) as ex vivo (spectrophotometric assay) can be utilized while optimizing existing regimens of PDT with the purpose to increase safety and efficacy of treatment as well as for the development of new PDT protocols with Photolon applied for new indications. Parameters of pharmacokinetics and biodistribution of Photolon/Fotolon as well as its' ability to cross an intact blood-brain barrier, are optimal for the majority of modern clinical applications of PDT. PMID- 19683211 TI - In vitro efficiency and mechanistic role of indocyanine green as photodynamic therapy agent for human melanoma. AB - BACKGROUND: Photodynamic therapy (PDT) is a promising treatment for superficial cancer. However, poor therapeutic results have been reported for melanoma, due to the high melanin content. Indocyanine green (ICG) has near infrared absorption (700-800 nm) and melanins do not absorb strongly in this area. This study explores the efficiency of ICG as a PDT agent for human melanoma, and its mechanistic role in the cell death pathway. METHODS: Human skin melanoma cells (Sk-Mel-28) were incubated with ICG and exposed to a low power Ti:Sapphire laser. Synchrotron-assisted Fourier transform infrared microspectroscopy and hierarchical cluster analysis were used to assess the cell damage and changes in lipid, protein, and nucleic acids. The cell death pathway was determined by analysis of cell viability and apoptosis and necrosis markers. RESULTS: In the cell death pathway, (1)O(2) generation evoked rapid multiple consequences that trigger apoptosis after laser exposure for only 15 min including the release of cytochrome c, the activation of total caspases, caspase-3, and caspase-9, the inhibition of NF-kappaB P65, and the enhancement of DNA fragmentation, and histone acetylation. CONCLUSION: ICG/PDT can efficiently and rapidly induce apoptosis in human melanoma cells and it can be considered as a new therapeutic approach for topical treatment of melanoma. PMID- 19683212 TI - Photo-toxic effects of 809-nm diode laser and indocyanine green on MDA-MB231 breast cancer cells. AB - Photodynamic therapy (PDT), due to its positive outcomes in clinical applications, easiness of practice and few side effects, is a good candidate for an efficient treatment of cancer. Indocyanine green (ICG), a water-soluble, anionic tricarbocyanine and non-toxic molecule is a promising photosensitive agent for PDT applications on tumor cells. ICG exhibits strong maximum absorption at around 805 nm which will be an advantage for its use in PDT; light at that wavelength can be used to treat deeper tumors. In this study the inhibitory growth effects of ICG-PDT on MDA-MB231 human breast cancer cells were investigated in a time course experiment. Cells were irradiated with a continuous wave diode laser (lambda=809 nm, 60 mW, 24 J cm(-2)). Cell viability was measured by MTT assay 0, 3, 6, 9, 12, 24 and 48h after light irradiation. The results showed that ICG-PDT application exerted its photo-oxidative effect on MDA-MB231 breast cancer cells immediately. Relative cell viability was determined throughout the 48h time course, and a consistent decrease was observed after ICG PDT applications. In conclusion, ICG when used in combination with near-infrared light showed a very fast (within 3h) and persistent (up to 48h) photo-toxic effect on MDA-MB231 human breast cancer cells. PMID- 19683213 TI - Induction of early apoptosis in human nasopharyngeal carcinoma cells by mTHPC mediated photocytotoxicity. AB - OBJECTIVES: In this study, the early apoptotic events elicited by mTHPC-mediated photo-cytotoxicity were explored in a human nasopharyngeal carcinoma cell line (NPC/HK1). METHODS AND MATERIALS: NPC/HK1 cells (5 x 10(3)) were incubated with photosensitizer mTHPC (0.8 microg/ml) in chamber slides for 20h and subjected to light irradiation at 2J/cm(2) (LD(80)). Morphologic changes of treated cells were examined under light microscopy and confocal microscopy at 0-4h after the light irradiation. The early stage of apoptosis was detected by fluorescein-conjugated Annexin V (Annexin V-FITC) assay. Mitochondrial membrane damage and cytochrome c release were determined by flowcytometric analysis. Bcl-2 expression was measured by Western blot analysis. RESULTS: One hour after mTHPC-mediated photodynamic therapy (PDT), microscopic examination showed membrane blebbing and cell shrinkage. Annexin V-FITC assay showed that a considerable number of NPC/HK1 cells became apoptotic. Flowcytometric analysis showed that the cytochrome c was released at 1h after PDT. Bcl-2 expression also declined significantly compared to control groups. CONCLUSIONS: mTHPC-mediated photo-cytotoxicity can effectively induce early apoptotic responses in NPC/HK1 cells which might be modulated by mitochondrial damages and Bcl-2 inhibition. PMID- 19683215 TI - PDT frontiers. PMID- 19683214 TI - Terahertz pulsed imaging--a potential medical imaging modality? AB - Terahertz imaging has progressed significantly over the last decade and there is now a significant body of research in its application to biomedical problems with the possibility of developing it into viable medical imaging modality in the future. The motivation being to fill some of the shortfalls in existing medical imaging technologies especially in detecting early stage cancers. We review the main developments in terahertz imaging to-date and highlight the most promising current areas of biomedical terahertz research. Additionally, we provide an overview of the principles behind terahertz imaging along with illustrated examples to aid understanding for those new to the technology. Our aim is to increase awareness of the existence and potential of the technology and inspire solutions to the remaining challenges in developing terahertz imaging into a novel medical imaging modality. PMID- 19683216 TI - Annual Scientific Meeting of British Medical Laser Association and UK-PDPDT interest group, 14 and 15 May 2009, Salisbury, UK. PMID- 19683217 TI - 12th World Congress of the International Photodynamic Association (IPA), 11-15th June 2009, Seattle, USA. PMID- 19683218 TI - Investigating mental representation of order with a speeded probed recall task. AB - We compare three models of representation of item order in a verbal STM task: item-item associations, item-position associations, and primacy gradient. A speeded probed recall task is used, in which a list of words is presented, immediately followed by a probe; participants must report as fast as possible the word that was in the probed position. In the number probe condition, a digit is presented and one must say the word in that position. In the word probe condition, the probe is an item of the list and participants must say the immediately following item. Response times (RTs) are analyzed according to probe type and position. The three models imply different predictions about RTs as a function of serial order in the two conditions. Our results suggest a serial, self-terminating search from the beginning of the list to the target position, except for the final position, which is directly accessible. The item-item and item-position association models are ruled out; the primacy gradient model accounts satisfactorily for our results, except for the finding of a larger recency effect with a number probe. Alternative interpretations are also discussed. PMID- 19683219 TI - Rheology of emulsions. AB - The review is devoted to the historical and modern understanding of rheological properties of emulsions in a broad range of concentration. In the limiting case of dilute emulsions, the discussion is based on the analogy and differences in properties of suspensions and emulsions. For concentrated emulsions, the main peculiarities of their rheological behaviour are considered. Different approaches to understand the concentration dependencies of viscosity are presented and compared. The effects of non-Newtonian flow curves and the apparent transition to yielding with increasing concentration of the dispersed phase are discussed. The problem of droplet deformation in shear fields is touched. The highly concentrated emulsions (beyond the limit of closest packing of spherical particles) are treated as visco-plastic media, and the principle features of their rheology (elasticity, yielding, concentration and droplet size dependencies) are considered. A special attention is paid to the problem of shear stability of drops of an internal phase starting from the theory of the single drop behaviour, including approaches for the estimation of drops' stability in concentrated emulsions. Polymer blends are also treated as emulsions, though taking into account their peculiarities due to the coexistence of two interpenetrated phases. Different theoretical models of deformation of polymer drops were discussed bearing in mind the central goal of predictions of the visco elastic properties of emulsions as functions of the properties of individual components and the interfacial layer. The role of surfactants is discussed from the point of view of stability of emulsions in time and their special influence on the rheology of emulsions. PMID- 19683220 TI - Expression of typical calpains in mouse molar. AB - OBJECTIVE: Calpain is a calcium ion-dependent cysteine protease, consisting of two primary isoforms (calpain1/calpain2) which mediate crucial cellular functions. The activity of the calpains is tightly regulated by the endogenous inhibitor calpastatin. Calpains have been detected in several studies during the embryonic and foetal stages. The aim of this study is to investigate the temporal transition of typical calpains and their inhibitor calpastatin during odontogenesis. DESIGN: We used the first molar of foetal ICR mice from embryonic day (E) 14 to postnatal day (PN) 7. Using laser microdissection and semi quantitative real-time PCR, we investigated calpain1, calpain2 and calpastatin expressions in each enamel epithelium, inner enamel epithelium, stellate reticulum and outer enamel epithelium. RESULTS: We found calpain1 and calpain2 mRNA increased in the all enamel epithelia between E18 and PN1. In addition calpastatin mRNA expression increased in the ameloblasts from PN1 to PN7. The immunohistochemistry results demonstrated that calpain1/calpain2 was present in the distal side of ameloblasts from PN1 to PN7, and calpastatin was present in the extracellular enamel matrix from E16 to PN1. Furthermore calpain1/calpain2 was present in the dentin, and calpastatin was detected in dentin producing odontoblasts and predentin at PN7. CONCLUSIONS: In this study the temporal transition of calpain1, calpain2 and calpastatin mRNA and the immunolocalization are identified during tooth development. Our results indicate that the calcium dependent proteases may play an important role in mouse molar development and extracellular calpain and calpastatin may be involved in molar mineralization. PMID- 19683221 TI - Incremental specificity of disgust sensitivity in the prediction of obsessive compulsive disorder symptoms: Cross-sectional and prospective approaches. AB - The present study examines the association between disgust sensitivity (DS) and obsessive-compulsive disorder (OCD) symptoms in two non-clinical samples. Findings from Study 1 (n=270) revealed a significant association between DS and OCD symptoms even after controlling for negative affect and anxiety sensitivity. Subsequent analysis also revealed a specific association between DS and the washing subtype of OCD symptoms when controlling for other OCD symptom dimensions. DS did not significantly predict residual change in total symptoms of OCD over a 12-week period (n=300) when controlling for risk factors for anxiety disorder symptoms in general (e.g., negative affect, anxiety sensitivity) and OCD specifically (e.g., obsessive beliefs) in Study 2. However, exploratory analyses suggest that DS may be predictive of residual change in some OCD symptom subtypes but not others. Implications of these findings for future research on the role of disgust in OCD are discussed. PMID- 19683222 TI - [Metastasis in the thyroid gland]. PMID- 19683224 TI - [Vacuum assisted closure (VAC) in the treatment of advanced diabetic foot]. AB - INTRODUCTION: Deep diabetic foot lesions pose an enormous therapeutic problem. The purpose of this study was to present the experience of the use of vacuum assisted closure (VAC) in the treatment of advanced and complicated diabetic foot lesions. MATERIAL AND METHODS: Five cases of advanced diabetic foot that were treated with VAC were prospectively studied. Three patients were diagnosed with renal failure, including one with renal transplant, who were receiving immunosuppression therapy. Four patients had undergone local foot surgery. The foot lesions were classified as grade 3 or 4 according to the Wagner classification. In all patients extensive debridement was performed that resulted in open minor amputations in four cases and resection of the metatarsophalangeal joint in one case. The VAC was applied during the same procedure. The median follow-up period of the patients was 9 months. RESULTS: Foot salvage was achieved in all cases. The median number of changes of VAC was 16 within median period of 8 weeks. Half of the changes were performed as an outpatient procedure. There were no major complications or clinical signs of infection observed. In one case before treatment with VAC began, angioplasty of the iliac artery and superficial femoral artery was performed. Other interventions carried out after the treatment was started were, two distal revascularizations and two partial transmetatarsal amputations. CONCLUSIONS: VAC appears to be very useful in the treatment of advanced diabetic foot lesions. PMID- 19683223 TI - [Prognostic factors of colorectal cancer liver metastasis after hepatic resection: Is this a uniform series? Are we talking of the same cases?]. PMID- 19683225 TI - Functional neuroanatomy of the encoding and retrieval processes of verbal episodic memory in MCI. AB - INTRODUCTION: The goal of this study was to explore the association between disease severity and performance on brain activation associated with episodic memory encoding and retrieval in persons with mild cognitive impairment (MCI). METHOD: This was achieved by scanning 12 MCI persons and 10 age- and education matched healthy controls while encoding words and while retrieving them in a recognition test. RESULTS: Behaviorally, there was no significant group difference on recognition performance. However, MCI and healthy controls showed different patterns of cerebral activation during encoding. While most of these differences demonstrated reduced activation in the MCI group, there were areas of increased activation in the left ventrolateral prefrontal cortex. Reduced activation was found in brain areas known to be either structurally compromised or hypometabolic in Alzheimer's disease (AD). In contrast, very few group differences were associated with retrieval. Correlation analyses indicated that increased disease severity, as measured with the Mattis Dementia Rating Scale, was associated with smaller activation of the right middle and superior temporal gyri. In contrast, recognition success in MCI persons was associated with larger activation of the left ventrolateral prefrontal cortex during the encoding phase. CONCLUSION: Overall, our results indicate that most of the memory-related cerebral network changes in MCI persons occur during the encoding phase. They also suggest that a prefrontal compensatory mechanism could occur in parallel with the disease-associated reduction of cerebral activation in temporal areas. PMID- 19683226 TI - Prenatal exposure to PCDDs/PCDFs and dioxin-like PCBs in relation to birth weight. AB - Several human studies have shown that low-level exposure to environmental contaminants, such as polychlorinated biphenyls (PCBs) and organochlorine pesticides, negatively influences birth outcomes. However, the effects of low level exposure to polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and dioxin-like PCBs (DL-PCBs) on birth outcomes have not been clarified in human studies. A prospective cohort study was established to investigate the possible adverse effects of PCDDs/PCDFs and DL-PCBs on fetal growth and neurodevelopment. We recruited 514 pregnant women between July 2002 and October 2005 in Sapporo, Japan. We measured 29 congener levels of PCDDs/PCDFs and DL-PCBs in maternal blood. Using multiple liner regression analysis of the association between birth weight and the levels of PCDDs/PCDFs and DL-PCBs with full adjustments for potential confounders, a significant adverse effect was observed regarding total PCDDs toxic equivalents (TEQ) levels (adjusted beta= 231.5g, 95% CI: -417.4 to -45.6) and total PCDFs TEQ levels (adjusted beta= 258.8g, 95% CI: -445.7 to -71.8). Among male infants, significant adverse associations with birth weight were found for total PCDDs TEQ level, total PCDDs/PCDFs TEQ level, and total TEQ level. However, among female infants, these significant adverse associations were not found. With regard to individual congeners of PCDDs/PCDFs and DL-PCBs, we found significantly negative association with the levels of 2,3,4,7,8-PeCDF (adjusted beta=-24.5g, 95% CI: -387.4 to 61.5). Our findings suggest that prenatal low-level exposure to PCDDs and PCDFs, especially 2,3,4,7,8-PeCDF, may accumulate in the placenta and retard important placental functions, which result in lower birth weight. PMID- 19683227 TI - PPAR-gamma expression in peritoneal endometriotic lesions correlates with pain experienced by patients. AB - Endometriosis is a significant gynecologic condition that can cause both pain and infertility and affects up to 15% of women during their reproductive years. In peritoneal endometriotic lesions, the expression of peroxisome proliferation activated receptor gamma, a nuclear receptor with antiinflammatory and neuroprotective roles, is positively correlated with the pain reported by patients. PMID- 19683228 TI - Blood group and ovarian hyperstimulation syndrome. AB - Blood group does not constitute a risk factor for the onset of the early form of ovarian hyperstimulation syndrome in women "at risk." PMID- 19683229 TI - Factors affecting thawed oocyte viability suggest a customized policy of embryo transfer. AB - OBJECTIVE: To identify factors that might affect the clinical outcome of oocyte slow freezing. DESIGN: Retrospective study. SETTING: Reproductive Medicine Unit, Italian Society for the Study of Reproductive Medicine, Bologna, Italy. PATIENT(S): Patients with spare metaphase II cryopreserved oocytes performing 371 thawing cycles. INTERVENTION(S): Oocytes were cryopreserved by slow freezing<40 hours after hCG administration (group A) and >or=40 hours after hCG administration (group B). Thawed oocytes were inseminated by intracytoplasmic sperm injection. MAIN OUTCOME MEASURE(S): Clinical pregnancy, implantation, abortion, and delivery rates. RESULT(S): Clinical pregnancy rate per thawed cycle (PR) and implantation rate (IR) were significantly higher in group A compared with group B both in young (PR: 25% vs. 9.6%; IR: 18.9% vs. 8.8%) and in older patients (PR: 25% vs. 10.1%; IR: 17.5% vs. 6.7%). In the young patient subgroup, clinical pregnancy and implantation rates with three transferred embryos were higher in group A vs. group B (PR: 72.7% vs. 25%, and IR: 36.4% vs. 12.5%, respectively). This difference was not found in the subgroup of older patients. CONCLUSION(S): The timing at which oocyte cryopreservation is performed and the number of transferred embryos play a key role in the clinical outcome. The suggested cut-off time for cryopreservation is between 39 and 40 hours after hCG administration. PMID- 19683230 TI - Similar ongoing pregnancy rates after blastocyst transfer in fresh donor cycles and autologous cycles using cryopreserved bipronuclear oocytes suggest similar viability of transferred blastocysts. AB - This retrospective cohort study compared blastocyst transfers in 136 fresh oocyte donor cycles and 69 autologous cycles using blastocysts derived from culture of thawed bipronuclear oocytes, all with oocytes derived from patients or donors less than 35 years old. The autologous cycles and oocyte donor cycles had similar rates of implantation (65.9% vs. 62.1%, respectively) and ongoing pregnancy (79.7% vs. 75.0%, respectively), suggesting that autologous blastocysts transferred after post-thaw extended culture have viability and implantation potential that are comparable with those of blastocysts transferred in fresh oocyte donor cycles. PMID- 19683231 TI - Coenzyme Q10 treatment reduces lipid peroxidation, inducible and endothelial nitric oxide synthases, and germ cell-specific apoptosis in a rat model of testicular ischemia/reperfusion injury. AB - In this experimental study, we assessed the preventive effects of coenzyme Q(10) (CoQ(10)) in a rat model of ischemia/reperfusion injury. The results of this study show that CoQ(10) administration before the reperfusion period of testicular torsion provides a significant decrease in testicular lipid peroxidation products and expressions of inducible nitric oxide synthase, endothelial nitric oxide synthase, and germ cell-specific apoptosis. PMID- 19683233 TI - Ovarian tissue thawing: a comparison of two conditions. AB - Two different protocols to thaw cryopreserved human ovarian cortex have been evaluated using a histological analysis. The slower one, based on a progressive dilution of cryoprotectants, seems to maintain an optimal follicle morphology. PMID- 19683232 TI - Analysis of pain and satisfaction with office-based hysteroscopic sterilization. AB - OBJECTIVE: To assess pain and patient satisfaction with office-based hysteroscopic sterilization. DESIGN: This prospective, observational study was designed to assess patient pain perception and satisfaction with office-based hysteroscopic sterilization using the Essure device (Conceptus, Mountain View, CA). SETTING: Faculty practice office at an inner-city urban medical center. PATIENT(S): Women seeking hysteroscopic sterilization. INTERVENTION(S): Office hysteroscopic sterilization under local anesthesia. MAIN OUTCOME MEASURE(S): Pain assessed at the time of the procedure by a 0-10 visual scale and satisfaction by a 1-5 scale. RESULT(S): From June 2003 to June 2006, 209 patients were recruited. The mean scores for average procedural pain, most procedural pain, and average menstrual pain were 2.6+/-2.1, 3.3+/-2.5, and 3.6+/-2.6, respectively. Standardized pain scores revealed that 149 subjects (70%) experienced average pain that was less than or equal to the pain experienced with their menses. Mean satisfaction rating for the procedure was 4.7+/-0.71. CONCLUSION(S): Office-based hysteroscopic sterilization performed with local anesthesia alone is well tolerated, and patients are satisfied with this method for permanent sterilization. PMID- 19683234 TI - Identification of uncertain nonlinear systems for robust fuzzy control. AB - In this paper, we consider fuzzy identification of uncertain nonlinear systems in Takagi-Sugeno (T-S) form for the purpose of robust fuzzy control design. The uncertain nonlinear system is represented using a fuzzy function having constant matrices and time varying uncertain matrices that describe the nominal model and the uncertainty in the nonlinear system respectively. The suggested method is based on linear programming approach and it comprises the identification of the nominal model and the bounds of the uncertain matrices and then expressing the uncertain matrices into uncertain norm bounded matrices accompanied by constant matrices. It has been observed that our method yields less conservative results than the other existing method proposed by Skrjanc et al. (2005). With the obtained fuzzy model, we showed the robust stability condition which provides a basis for different robust fuzzy control design. Finally, different simulation examples are presented for identification and control of uncertain nonlinear systems to illustrate the utility of our proposed identification method for robust fuzzy control. PMID- 19683235 TI - Treatment of long bone aseptic non-unions: monotherapy or polytherapy? PMID- 19683236 TI - A pathobiologic link between risk factors profile and morphological markers of carotid instability. AB - OBJECTIVE: Although cardiovascular risk factors have been strongly linked to carotid intimal-media thickness, their association with plaque progression towards instability is poorly understood. We evaluated a large database of endarterectomy specimens removed from symptomatic and asymptomatic patients to determine the correlation between major cardiovascular risk factors and carotid plaque morphology. METHODS: Incidence of thrombotic, vulnerable and stable plaques together with the degree of plaque inflammatory infiltration was evaluated in 457 carotid atherosclerotic lesions. Clinical records were reviewed in all cases for risk factors profile. RESULTS: Thrombotic plaques were more frequently observed in patients affected by stroke (66.9%) as compared to TIA (36.1%) and asymptomatic patients (26.8%, p<0.001). Out of 457 carotid plaques removed during carotid endarterectomy, 181 (39.6%) were represented by thrombotic plaques, 72 (15.8%) by vulnerable plaques (thin cap fibroateroma) and 204 (44.6%) by stable plaques. At the multivariate analysis, a strong association was observed between hypertension, low HDL-cholesterol (HDL-C) and ratio of total to HDL-C >5 with vulnerable and thrombotic carotid plaques. Hypertension (p=0.001), hypercholesterolemia (p=0.05) and low HDL-C (p=0.001) significantly also correlated with the presence of high inflammatory infiltrate of the plaque. When multivariate analysis was restricted to asymptomatic patients, hypertension (p=0.009, OR 2.29), low HDL-cholesterol (p=0.01 OR 2.21) and the ratio of total to HDL-C >5 (p=0.03, OR 2.07) were confirmed to be the risk factors most significantly associated to unstable plaques. The relative risk to carry an unstable plaque for asymptomatic patients with high Framingham Risk Score as compared with those with low risk score was 2.06 (95% C.I., 1.26-3.36). CONCLUSIONS: The present histopathological study identifies risk factors predictive of increased risk of carotid plaque rupture and thrombosis. Asymptomatic patients with high risk factors profile may constitute a specific target to reduce the likelihood of cerebrovascular accidents even in the presence of non-flow-limiting plaque. PMID- 19683237 TI - Lysyl oxidase resolves inflammation by reducing monocyte chemoattractant protein 1 in abdominal aortic aneurysm. AB - Lysyl oxidase (LOX) is an enzyme critical for the stability of extracellular matrix and also known to have diverse biological functions. Little is known, however, about the role of LOX in regulating inflammation. Here we demonstrate that LOX suppresses secretion of monocyte chemoattractant protein-1 (MCP-1) in cultured vascular smooth muscle cells. Furthermore, enhancement of LOX activity reduces MCP-1 in a mouse model of abdominal aortic aneurysm (AAA), thereby preventing macrophage infiltration and AAA progression. These findings suggest that LOX has a novel function in resolving inflammation by reducing MCP-1 in AAA. PMID- 19683239 TI - Down-regulation of RhoA is involved in the cytotoxic action of lipophilic statins in HepG2 cells. AB - OBJECTIVE: Hepatotoxicity is one of the major complaints that occur during lipid lowering therapy with 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase inhibitors, known as statins. We reported earlier that lipophilic but not hydrophilic statins induce apoptosis through inhibition of mevalonate biosynthesis cascade in Chang liver cells. The present study was designed to determine the role for small G protein RhoA in the hepatocytotoxicity of statins. METHODS: Statin-induced hepatocytotoxicity in HepG2 cells were assessed by WST-8 cell viability assay, JC-1 mitochondrial membrane potential assay and caspase-3/7 activity assay. Cytosolic RhoA was detected by Western blotting and RhoA activation was measured by ELISA. RESULTS: The lipophilic atorvastatin but not the hydrophilic pravastatin induced the mitochondrial membrane depolarization and the activation of caspase-3/7, which led to cell injury. Supplementation of mevalonate or geranylgeranyl pyrophosphate (GGPP) but not farnesyl pyrophosphate (FPP) reversed these cellular events and cell death induced by atorvastatin. Atorvastatin induced a translocation of RhoA protein into the cytosol and inhibited the activity of the protein. In addition, atorvastatin reduced mitochondrial membrane potential, which was mimicked by GGTase inhibitor GGTI 2147 or the specific RhoA inhibitor such as toxin B and C3 exoenzyme. However, only a few cells revealed mitochondrial membrane depolarization and a loss of viability after exposure to the Rho-kinase inhibitors such as Y-27632 and hydroxy fasudil. CONCLUSIONS: RhoA inactivation and to a lesser extent Rho-kinase inhibition after depletion of GGPP is implicated in the etiology of mitochondrial membrane depolarization and subsequent caspase-dependent cell death induced by the lipophilic statin in HepG2 cells. PMID- 19683238 TI - Coronary late lumen loss of drug eluting stents is associated with increased serum levels of the complement components C3a and C5a. AB - OBJECTIVE: Drug eluting stents (DES) reduce recurrent luminal narrowing through anti-migratory and anti-proliferative effects. However, recent concerns arose that DES may also induce significant chronic inflammatory responses that may impair vascular healing and lead to in-stent restenosis (ISR). As the complement components C3a and C5a exert particularly strong chemotactic and proinflammatory effects, we examined the association of serum levels of C3a and C5a and ISR after implantation of DES. METHODS: We included 82 patients that were treated with 151 DES. Blood samples were taken directly before and 24h after PCI. Serum levels of C3a and C5a were measured by specific ELISA and restenosis was evaluated at 6-8 months by coronary angiography. RESULTS: C5a but not C3a increased after implantation of DES (p<0.05). During the follow-up period, two patients (2.4%) died of cardiovascular causes and 12 patients (7.9% of stents, 15% of patients) developed ISR. Serum levels of C3a before and 24h after PCI as well as C5a levels at baseline were significantly higher in patients that developed ISR at follow up. C3a and C5a at baseline were significantly associated to angiographic late lumen loss independent from clinical and procedural risk factors. CONCLUSION: Increased complement activation as measured by higher levels of C3a and C5a before PCI is significantly associated with late lumen loss. Inhibition of the complement cascade to prevent ISR warrants further investigation. PMID- 19683240 TI - The consistency of maximum running speed measurements in humans using a feedback controlled treadmill, and a comparison with maximum attainable speed during overground locomotion. AB - Consistent measurement of maximum running speed overground is problematic due to the difficulty in precise, continual measurement of speed, and the substantial workload in accelerating the body promoting the onset of fatigue. Treadmills remove the requirement for acceleration which enables more repeats. They also allow experiments to be carried out in controlled environments and where space is limited, but they usually depend on manual and subjective speed control. Here we used a draw-wire position sensor and a proportional-derivative (PD) controller to automatically adjust treadmill belt speed of a large equine treadmill. The feedback loop took the real-time position and velocity of the runner relative to the front of the treadmill as input. This control system allowed runners to accelerate from walking speed to a peak running speed within a few strides and then decelerate as quickly as they wished. We used the system to evaluate the variation in maximum speed determination that results from one trial to 10 trials, in eleven individuals. Three trials gave a maximum speed 97.8% of that achieved after ten. The approach used is appropriate for any treadmill where the running zone length is greater than three metres and the speed controller can be externally controlled. Subjects ran 11.5% faster on the treadmill than overground, part of which can be explained by the removal of aerodynamic drag and the fatigue of overground running. Additional factors may, however, contribute to athletes running faster on a treadmill, for instance some aspect of stability or control. PMID- 19683241 TI - Simultaneous identification of glucosinolates and phenolic compounds in a representative collection of vegetable Brassica rapa. AB - Brassica raparapa group is widely distributed and consumed in northwestern Spain. The consumption of Brassica vegetables has been related to human health due to their phytochemicals, such as glucosinolates and phenolic compounds that induce a variety of physiological functions including antioxidant activity, enzymes regulation and apoptosis control and the cell cycle. For first time in Brassica crops, intact glucosinolates and phenolic compounds were simultaneously identified and characterized. Twelve intact glucosinolates, belonging to the three chemical classes, and more than 30 phenolic compounds were found in B. rapa leaves and young shoots (turnip greens and turnip tops) by LC-UV photodiode array detection (PAD)-electrospray ionization (ESI). The main naturally occurring phenolic compounds identified were flavonoids and derivatives of hydroxycinnamic acids. The majority of the flavonoids were kaempferol, quercetin and isorhamnetin glycosylated and acylated with different hydroxycinnamic acids. Quantification of the main compounds by HPLC-PAD showed significant differences for most of compounds between plant organs. Total glucosinolate content value was 26.84 micromol g(-1) dw for turnip greens and 29.11 micromol g(-1) dw for turnip tops; gluconapin being the predominant glucosinolate (23.2 micromol g(-1) dw). Phenolic compounds were higher in turnip greens 51.71 micromol g(-1) dw than in turnip tops 38.99 micromol g(-1) dw, in which flavonols were always the major compounds. PMID- 19683242 TI - Application of histograms in evaluation of large collections of gas chromatographic retention indices. AB - The effective use of gas chromatographic retention data presented in the form of retention indices (RI) requires the development of a comprehensive structure based digital archive of retention parameters. Development of such an archive includes the collection of all available RI values for a variety of compounds including replicates measured under slightly different conditions. Review of retention data often shows a relatively wide range of RI values for certain well studied compounds that is larger than expected on the basis of the simple reproducibility of experimental measurements. The finding of unusual RI data distributions and their examination presents a possible way to detect and correct errors during the development of comprehensive RI libraries. Our approach involves the construction of histograms representing the distribution of data points in various RI intervals. The observed shape of the distribution is compared to the expected and observed shapes for well-identified compounds. Significant systematic deviations represent anomalies in the sets of RI data. The occurrence of more than a single maximum on a histogram generally indicates the presence of erroneous data. For some compounds such multimode RI distributions may be caused by differences in experimental conditions of the RI determination. The construction and interpretation of histograms for compounds with multiple RI measurements is illustrated by several examples. Thus, the RI sub-set for the diterpene alcohol, isophytol, was separated from the RI data set for phytol and four additional sub-groups of published RI data for one of the sesquiterpenes, gamma-elemene, were re-identified as alpha-, beta-, delta-elemenes and germacrene B. PMID- 19683243 TI - Improvement of separation efficiencies of anion-exchange chromatography using monolithic silica capillary columns modified with polyacrylates and polymethacrylates containing tertiary amino or quaternary ammonium groups. AB - Anion-exchange (AEX) columns were prepared by on-column polymerization of acrylates and methacrylates containing tertiary amino or quaternary ammonium groups on monolithic silica in a fused silica capillary modified with anchor groups. The columns provided a plate height (H) of less than 10 microm at optimum linear velocity (u) with keeping their high permeability (K=9-12 x 10(-14) m2). Among seven kinds of AEX columns, a monolithic silica column modified with poly(2 hydroxy-3-(4-methylpiperazin-1-yl)propyl methacrylates) (HMPMA) showed larger retentions and better selectivities for nucleotides and inorganic anions than the others. The HMPMA column of 410 mm length produced 42,000-55,000 theoretical plates (N) at a linear velocity of 0.97 mm/s with a backpressure of 3.8 MPa. The same column could be employed for a fast separation of inorganic anions in 1.8 min at a linear velocity of 5.3 mm/s with a backpressure of 20 MPa. In terms of van Deemter plot and separation impedance, the HMPMA column showed higher performance than a conventional particle-packed AEX column. The HMPMA column showed good recovery of a protein, trypsin inhibitor, and it was applied to the separation of proteins and tryptic digest of bovine serum albumin (BSA) in a gradient elution, to provide better separation compared to a conventional particle-packed AEX column. PMID- 19683244 TI - Application of retention modelling to the simulation of separation of organic anions in suppressed ion chromatography. AB - The ion-exchange separation of organic anions of varying molecular mass has been demonstrated using ion chromatography with isocratic, gradient and multi-step eluent profiles on commercially available columns with UV detection. A retention model derived previously for inorganic ions and based solely on electrostatic interactions between the analytes and the stationary phase was applied. This model was found to accurately describe the observed elution of all the anions under isocratic, gradient and multi-step eluent conditions. Hydrophobic interactions, although likely to be present to varying degrees, did not limit the applicability of the ion-exchange retention model. Various instrumental configurations were investigated to overcome problems associated with the use of organic modifiers in the eluent which caused compatibility issues with the electrolytically derived, and subsequently suppressed, eluent. The preferred configuration allowed the organic modifier stream to bypass the eluent generator, followed by subsequent mixing before entering the injection valve and column. Accurate elution prediction was achieved even when using 5-step eluent profiles with errors in retention time generally being less than 1% relative standard deviation (RSD) and all being less than 5% RSD. Peak widths for linear gradient separations were also modelled and showed good agreement with experimentally determined values. PMID- 19683246 TI - Sub-micrometer precision of optical imaging to locate the free surface of a micrometer fluid shape. AB - In this note, we explore the precision of the optical imaging method for measuring the free surface position of a micrometer fluid shape. For this purpose, images of a liquid film deposited on a rod were acquired and processed. The resulting contour was compared with the corresponding solution to the Young Laplace equation. The average deviation was about 30nm, 25 times smaller than the pixel size, reflecting the validity of optical imaging for most applications in microfluidics. PMID- 19683245 TI - Liquid chromatography-tandem mass spectrometry reveals the widespread occurrence of flavonoid glycosides in honey, and their potential as floral origin markers. AB - HPLC-MS-MS analysis of unifloral honey extracts has shown the occurrence of flavonoid glycosides in most of the analyzed samples. These compounds are not present in large amounts, but can reach up to 600 microg/100g honey in canola and rapeseed honeys. Rhamnosyl-hexosides (tentatively rutinosides and neohesperidosides) and dihexosides (hexosyl(1-->2)hexosides and hexosyl(1- >6)hexosides) of flavonols such as quercetin, kaempferol, isorhamnetin and 8 methoxykaempferol, are the main flavonoid glycosides found in honey. However, flavonoid triglycosides and monoglycosides are also detected in some floral origins. Eucalyptus and orange blossom nectars were collected and analyzed showing that nectar flavonoid glucosides, as is the case of eucalyptus flavonoids, can be readily hydrolyzed by the bee saliva enzymes, while flavonoid rhamnosyl-glucosides, as is the case of citrus nectar flavonoids, are not hydrolyzed, and because of these reasons the flavonoid glycoside content of citrus honey is higher than that of eucalyptus honey that contains mainly aglycones. The flavonoid glycoside profiles detected in honeys suggest that this could be related to their floral origin and the results show that the HPLC-MSn ion trap analysis of flavonoid glycosides in honey is a promising analytical method to help in the objective determination of the floral origin of unifloral honeys. PMID- 19683247 TI - Expanding mesoporosity of triblock-copolymer-templated silica under weak synthesis acidity. AB - With initial aging at low temperature for enough time, silicas with large mesoporosity were synthesized using triblock copolymer as template agent under weak acidities. SBA-15 with periodic mesostructure and short mesochannels could be synthesized at pH 2.5-3.0 within weak acidity range, and the surface areas, pore diameters and pore volumes reached up to ca. 1000m(2)/g, 8.8nm and 2.0cm(3)/g, respectively, which were significantly higher than those of the conventional SBA-15 synthesized under strong acidities. Mesoporous silica with wormhole structure and abundant textural porosity was formed at pH approximately 3.5. The increased hydrophobic volume of the copolymer micelles at elevated pH values was responsible for the enlargement of mesoporosity in the products. The materials synthesized under weak acidities showed lower hexagonal ordering in comparison to the general SBA-15 synthesized under strong acidities because the decreased hydronium ion concentration induced relatively weaker assembly forces during the synthesis. Nonetheless, the short mesochannels and large pore diameter in the products might be beneficial to some applications in which fast diffusion of molecules is required. PMID- 19683248 TI - Comparison of molecular dynamics simulations with triple layer and modified Gouy Chapman models in a 0.1M NaCl-montmorillonite system. AB - Molecular dynamics (MD) simulations of a montmorillonite/water interface at the pore scale were carried out at 0.1molL(-1) NaCl concentration in order to constrain cation, anion, and water distribution and mobility influenced by the mineral surface. MD results enabled anion exclusion and cation condensation at the surface to be quantified. MD-derived values could then be compared with macroscopic model results obtained from the Modified Gouy-Chapman (MGC) theory. While the Na concentration profile is well reproduced in the diffuse layer, anion exclusion is overestimated by the MGC theory under our experimental conditions. We also showed that MD simulations can be used to constrain Basic Stern model parameters or, in combination with zeta potential measurements, can be used to constrain triple layer model (TLM) parameters by providing suitable values for the capacitance values. Na sorption intrinsic equilibrium constant values for clay basal surfaces are given accordingly. PMID- 19683249 TI - Differential transport and dispersion of colloids relative to solutes in single fractures. AB - This work employed numerical experiments simulating colloid and solute transport in single parallel-plate fractures, using the random walk particle tracking method, to demonstrate that (1) there exists an aspect ratio of the colloid radius to half the fracture aperture, delta(o), where the average velocities of colloids and solutes are similar. When delta>delta(o), the velocity distribution assumption is satisfied, and the fact that the ratio of the colloid transport velocity to the solute transport velocity, tau(p), decreases as delta increases is well documented in the literature. However, when delta or = 6 high). RESULTS: Most children had one SABA prescriber (62%); 13% had multiple prescribers in the same practice as the primary care provider and 25% had multiple prescribers in different practices. Children with multiple prescribers in different practices had increased odds of an emergency department visit compared with those with 1 prescriber, among those with high SABA prescription frequency (AOR: 2.7, 95% CI: 1.9, 3.9), as well as those with low prescription frequency (AOR: 1.7, 95% CI: 1.3, 2.2). CONCLUSIONS: Children with discontinuity of SABA prescribers have an increased risk of asthma emergency department visits, irrespective of their SABA prescription frequency. Primary care providers may have difficulty identifying patients at high risk with asthma solely on the basis of SABAs prescribed within their own practices. PMID- 19683254 TI - Childhood nephrotic syndrome in Cambodia: an association with gastrointestinal parasites. AB - OBJECTIVES: To describe childhood nephrotic syndrome (NS) in Cambodia and to evaluate whether initial presentation or relapse is associated with gastrointestinal parasitic infection. STUDY DESIGN: We reviewed the records of 112 children with NS. A retrospective cross-sectional study compared 99 stool exams from 63 children with NS with 12 365 stool exams from 9495 controls. RESULTS: The male-to-female ratio was 1.7; the mean age of presentation was 8.95 years--44% were hypertensive, 44% had microscopic hematuria, 40% had eosinophilia, and 41% had acute renal failure; 92.7% were steroid sensitive, 12.7% were steroid dependent, and 8.9% were frequent relapsers. Peritonitis and death were rare outcomes. Giardia lamblia (OR, 3.62; 95% CI, 2.0 to 6.1), Strongyloides stercoralis (OR, 3.59; 95% CI, 1.3 to 8.2), and Hookworm species (OR, 2.57; 95% CI, 1.0 to 5.5) were more likely to be isolated from the children with NS than the controls. CONCLUSIONS: The clinical course of childhood NS in Cambodia is similar to the developed world. Differences at presentation included older age and increased prevalence of microscopic hematuria, hypertension, eosinophilia, and acute renal failure. This study demonstrates an association between G lamblia, S stercoralis, and possibly Hookworm species and the onset of NS. PMID- 19683255 TI - Prolonged unconjugated hyperbiliriubinemia in breast-fed male infants with a mutation of uridine diphosphate-glucuronosyl transferase. AB - OBJECTIVE: To test the hypothesis that a mutation in uridine diphosphate glucuronosyl transferase 1A1 (UGT1A1) gene of breast-fed infants is a contributory factor to prolonged unconjugated hyperbilirubinemia. STUDY DESIGN: Of 125 breast-fed term infants, 35 infants had prolonged unconjugated hyperbilirubinemia; another 90 breast-fed neonates without prolonged jaundice were control infants. The polymerase chain reaction-restriction fragment length polymorphism method was used to detect the known variant sites (promoter area, nucleotides 211, 686, 1091, and 1456) of the UGT1A1 gene. RESULTS: Of 35 breast fed infants with prolonged unconjugated hyperbilirubinemia, 29 had at least 1 mutation of the UGT1A1 gene. Variation at nucleotide 211 was most common. The percentages of the neonates carrying the variant nucleotide 211 were significantly different between the prolonged hyperbilirubinemia group and control neonates. Male breast-fed infants had a higher risk than female infants for prolonged hyperbilirubinemia. CONCLUSIONS: Male breast-fed neonates with a variant nucleotide 211 in UGT1A1 have a high risk for developing prolonged hyperbilirubinemia. PMID- 19683257 TI - The natural history of Shwachman-Diamond syndrome-associated liver disease from childhood to adulthood. AB - OBJECTIVES: In order to characterize the natural course of Shwachman-Diamond syndrome (SDS)-associated hepatopathy we evaluated liver biochemistry and imaging findings, and their evolution with age, in patients with SDS and verified SBDS mutations. STUDY DESIGN: Retrospective and cross-sectional liver imaging, biochemical and histologic data of 12 patients (age range 2.1 to 37 years) with SBDS mutations were analyzed. Hepatic volume and parenchymal structure were determined from magnetic resonance imaging data. RESULTS: Hepatomegaly and aminotransaminase elevation was observed in most of the patients with SDS at an early age; values normalized by age 5 years and remained normal over extended follow-up. Mild to moderate serum bile acid elevation was noted in 7 patients (58%). On magnetic resonance imaging, no patients (n = 11) had evidence of hepatic steatosis, cirrhosis, or fibrosis. Three middle-aged patients had hepatic microcysts. CONCLUSIONS: SDS-associated hepatopathy has overall good prognosis. No major hepatic abnormalities developed during extended follow-up to adulthood. Mild cholestasis in follow-up even after normalization of transaminase levels may reflect primary alterations in liver metabolism in SDS. PMID- 19683256 TI - Gastric acid inhibition for fat malabsorption or gastroesophageal reflux disease in cystic fibrosis: longitudinal effect on bacterial colonization and pulmonary function. AB - OBJECTIVES: To investigate bacterial colonization and pulmonary function longitudinally in patients with cystic fibrosis (CF) receiving drugs for gastric acid (GA) inhibition for fat malabsorption or for gastroesophageal reflux disease (GERD). STUDY DESIGN: A retrospective cohort study of 218 pediatric patients with CF was performed. Multilevel modeling was used to perform longitudinal analysis of forced expiratory volume in 1 second (FEV(1)), forced vital capacity (FVC), maximum expiratory flow at 50% of FVC (MEF(50)), and maximal mid-expiratory flow between 25% and 75% of FVC (MMEF(25-75)). Cox regression was used to calculate Pseudomonas aeruginosa- and Staphylococcus aureus-free survival. RESULTS: Patients with CF and GA inhibition had a significantly smaller yearly decline of MEF(50) and MMEF(25-75) compared with control subjects. Other pulmonary function parameters and P aeruginosa or S aureus acquisition or colonization were not different from that of control subjects. GERD was associated with a significantly reduced pulmonary function (FEV(1) and FVC) and an earlier acquisition of P aeruginosa and S aureus. CONCLUSIONS: GA inhibition did not affect pulmonary function or bacterial acquisition and therefore is not contraindicated in patients with CF. GA inhibition might improve pulmonary function with time, because the decline of MEF(50) and MMEF(25-75) was less pronounced. GERD was associated with a reduced pulmonary function and an earlier acquisition of P aeruginosa and S aureus. Therefore the diagnosis and treatment of GERD should be aggressively pursued in patients with CF. PMID- 19683258 TI - Neuronal correlates and serotonergic modulation of behavioural inhibition and reward in healthy and antisocial individuals. AB - Individuals with antisocial personality disorder (ASPD) are impulsive and show impairment in reinforcement processing. There is increasing evidence for a neurobiological basis of psychopathy, which shares some of the characteristics of ASPD, but research on the neuronal correlates of neuropsychological processes in ASPD remains limited. Furthermore, no research has examined the effects of serotonergic manipulation on brain activations in antisocial groups. In this study, 25 male participants with ASPD (mean age 42.1) and 32 male control participants (mean age 30.5; 25 participants providing usable scans) were randomly allocated to receive the 5-HT(2C)-agonist mCPP or placebo. Participants were scanned using functional magnetic resonance imaging (fMRI) during a behavioural inhibition (Go/NoGo) and a reward task. In comparison to healthy controls the ASPD group showed reduced task related activations in the dorsolateral prefrontal cortex (DLPFC) but increased signal in the pre/subgenual anterior cingulate cortex (ACC) in the Go/No-Go task and increased activation in OFC in the reward task. mCPP modulated brain responses in both tasks in the whole group. Interactions between group and drug occured in bilateral OFC, caudate and ventral pallidum during the reward task but no significant interactions were found in the Go/No-Go task. This suggests that ASPD involves altered serotonin modulation of reward, but not motor inhibition pathways. These findings suggest that ASPD involves altered DLPFC, ACC and OFC function. Altered serotonergic modulation of reward pathways seen in the ASPD group raises the possibility that targeting serotonin systems may be therapeutic. PMID- 19683259 TI - Family psychiatric history, peritraumatic reactivity, and posttraumatic stress symptoms: a prospective study of police. AB - BACKGROUND: Family history of psychiatric and substance use disorders has been associated with posttraumatic stress disorder (PTSD) in cross-sectional studies. METHOD: Using a prospective design, we examined the relationships of family history of psychiatric and substance use disorders to posttraumatic stress symptoms in 278 healthy police recruits. During academy training, recruits were interviewed on family and personal psychopathology, prior cumulative civilian trauma exposure, and completed self-report questionnaires on nonspecific symptoms of distress and alcohol use. Twelve months after commencement of active duty, participants completed questionnaires on critical incident exposure over the previous year, peritraumatic distress to the worst critical incident during this time, and posttraumatic stress symptoms. RESULTS: A path model indicated: (1) family loading for mood and anxiety disorders had an indirect effect on posttraumatic stress symptoms at 12 months that was mediated through peritraumatic distress to the officer's self-identified worst critical incident, (2) family loading for substance use disorders also predicted posttraumatic stress symptoms at 12 months and this relationship was mediated through peritraumatic distress. CONCLUSION: These findings support a model in which family histories of psychopathology and substance abuse are pre-existing vulnerability factors for experiencing greater peritraumatic distress to critical incident exposure which, in turn, increases the risk for development of symptoms of posttraumatic stress disorder. Replication in other first responders, military and civilians will be important to determine generalizability of these findings. PMID- 19683261 TI - Prostatic transition zone directed needle biopsies uncommonly sample clinically relevant transition zone tumors. AB - PURPOSE: We compared prostate cancer detected in transition zone directed needle biopsies with those in corresponding radical prostatectomy specimens. MATERIALS AND METHODS: We reviewed needle biopsy and radical prostatectomy slides from 61 patients in whom cancer was present on transition zone directed needle biopsy. We assessed needle biopsy cancer features as well as transition zone lesions and dominant tumor sites on radical prostatectomy. RESULTS: Prostate cancer was detected in 25 of 61 left (41%), 23 of 61 right (38%) and 13 of 61 bilateral (21%) transition zone directed needle biopsies. On radical prostatectomy 24 of 61 cases (39.5%) had no tumor in the transition zone, 24 of 61 (39.5%) had nondominant transition zone cancer and 13 of 61 (21%) had a dominant transition zone lesion. Of cases with cancer in the left and right transition zone directed needle biopsy 18 of 38 (47%) and 17 of 36 (47%), respectively, had no transition zone tumor or showed tumor in the contralateral transition zone only at radical prostatectomy. In 8 cases the transition zone directed core was the only one with cancer on needle biopsy and 2 of 8 (25%) such cases showed dominant transition zone cancer at radical prostatectomy. CONCLUSIONS: Cancer identified in transition zone directed needle biopsy cores was not from the transition zone or did not reflect a dominant transition zone lesion in almost 80% of cases. Cancer identified in a left or right transition zone directed needle biopsy did not predict ipsilateral transition zone cancer in almost 50% of cases. These findings suggest that such biopsies do not adequately characterize transition zone tumors. Thus, care should be taken in their interpretation. PMID- 19683260 TI - Progranulin plasma levels as potential biomarker for the identification of GRN deletion carriers. A case with atypical onset as clinical amnestic Mild Cognitive Impairment converted to Alzheimer's disease. AB - Progranulin (GRN) mutations are associated with different clinical phenotypes, including Frontotemporal Lobar Degeneration (FTLD), Corticobasal Degeneration and Alzheimer's disease (AD). In addition, the range of age at onset is very wide and patients presenting initial symptoms around eighty years have been described. Previous studies demonstrated that progranulin plasma levels determination may be a reliable method to identify GRN deletion carriers. We thus evaluated progranulin plasma levels in all patients followed at our Alzheimer's Centre whose plasma was available (n=176) and found four patients displaying low values. Three of them carried the CACT deletion in exon 7 and their clinical diagnosis was behavioral variant Frontotemporal Dementia. We also identified a patient carrying a previously reported CAGT deletion in exon 5. Here, we report on this case. The onset of symptoms was at 77 years and the initial diagnosis was of amnestic Mild Cognitive Impairment (aMCI), which converted to AD six months later. In the following years, the patient also developed behavioral disturbances, gait apraxia and parkinsonian symptoms. At present, she is 84 years old and is still followed-up periodically. This case confirms progranulin plasma levels as a reliable biomarker to identify GRN deletion carriers and discriminate between FTLD and other dementias which may mimic it. We thus encourage the inclusion of this non-invasive and easy test in clinical practice. PMID- 19683262 TI - Image guided photothermal focal therapy for localized prostate cancer: phase I trial. AB - PURPOSE: We ascertained the feasibility and safety of image guided targeted photothermal focal therapy for localized prostate cancer. MATERIALS AND METHODS: Twelve patients with biopsy proven low risk prostate cancer underwent interstitial photothermal ablation of the cancer. The area of interest was confirmed and targeted using magnetic resonance imaging. Three-dimensional ultrasound was used to guide a laser to the magnetic resonance to ultrasound fused area of interest. Target ablation was monitored using thermal sensors and real-time Definity contrast enhanced ultrasound. Followup was performed with a combination of magnetic resonance imaging and prostate biopsy. Validated quality of life questionnaires were used to assess the effect on voiding symptoms and erectile function, and adverse events were solicited and recorded. RESULTS: Interstitial photothermal focal therapy was technically feasible to perform. Of the patients 75% were discharged home free from catheter the same day with the remainder discharged home the following day. The treatment created an identifiable hypovascular defect which coincided with the targeted prostatic lesion. There were no perioperative complications and minimal morbidity. All patients who were potent before the procedure maintained potency after the procedure. Continence levels were not compromised. Based on multicore total prostate biopsy at 6 months 67% of patients were free of tumor in the targeted area and 50% were free of disease. CONCLUSIONS: Image guided focal photothermal ablation of low risk and low volume prostate cancer is feasible. Early clinical, histological and magnetic resonance imaging responses suggest that the targeted region can be ablated with minimal adverse effects. It may represent an alternate treatment approach to observation or delayed standard therapy in carefully selected patients. Further trials are required to demonstrate the effectiveness of this treatment concept. PMID- 19683263 TI - Editorial comment. PMID- 19683264 TI - Editorial comment. PMID- 19683266 TI - Guideline for management of the clinical T1 renal mass. PMID- 19683267 TI - Editorial comment. PMID- 19683268 TI - Editorial comment. PMID- 19683269 TI - Optimizing prostate cancer detection: 8 versus 12-core biopsy protocol. AB - PURPOSE: We compared prostate cancer detection rates achieved using an 8 and 12 core biopsy protocol in a clinical population to determine the significance of additional transition zone sampling on repeat biopsy. MATERIALS AND METHODS: Between September 2004 and September 2007, 269 eligible patients with a clinical suspicion of prostate cancer referred to our department were randomized to an 8 core lateral (group 1) or a 12-core lateral and parasagittal (group 2) transrectal ultrasound guided prostate biopsy protocol. Study inclusion criteria were age dependent increased serum prostate specific antigen (1.25 ng/ml or greater at ages less than 50 years, 1.75 or greater at ages 50 to less than 60 years, 2.25 or greater at ages 60 to less than 70 years and 3.25 or greater at ages 70 years or greater), positive digital rectal examination and/or suspicious transrectal ultrasound. After negative first round biopsy patients underwent 12 core biopsy, including 4 transition zone cores. RESULTS: Nine patients were excluded from analysis because of protocol violation or they did not complete the whole biopsy procedure due to discomfort. The cancer detection rate in groups 1 and 2 did not differ significantly (34.1% or 45 of 132 patients and 38.3% or 49 of 128, respectively, p = 0.48). Detected cancer median Gleason scores were similar in the groups. Of 109 patients who underwent repeat biopsy prostate cancer was detected in 20 (14.4%), of whom 9 had positive cores from the transition zone and 6 had positive biopsies only from the transition zone. CONCLUSIONS: There are no statistically significant differences in the prostate cancer detection rate between 8 and 12-core prostate biopsy protocols. Transition zone biopsies contribute to prostate cancer detection in a repeat biopsy protocol. PMID- 19683270 TI - Optical biopsy of human bladder neoplasia with in vivo confocal laser endomicroscopy. AB - PURPOSE: Confocal laser endomicroscopy is a new endoscopic imaging technology that could complement white light cystoscopy by providing in vivo bladder histopathology. We evaluated confocal laser endomicroscopy by imaging normal, malignant appearing and indeterminate bladder mucosa in a pilot study. MATERIALS AND METHODS: Patients scheduled to undergo transurethral resection of bladder tumors were recruited during a 3-month period. After standard cystoscopy fluorescein was administered intravesically and/or intravenously as a contrast dye. A 2.6 mm probe based confocal laser endomicroscope was passed through a 26 Fr resectoscope to image normal and abnormal appearing areas. The images were collected with 488 nm excitation at 8 to 12 frames per second. The endomicroscopic images were compared with standard hematoxylin and eosin analysis of transurethral resection of bladder tumor specimens. RESULTS: Of the 27 recruited patients 8 had no cancer, 9 had low grade tumors, 9 had high grade tumors and 1 had a low grade tumor with a high grade focus. Endomicroscopic images demonstrated clear differences between normal mucosa, and low and high grade tumors. In normal urothelium larger umbrella cells are seen most superficially followed by smaller intermediate cells and the less cellular lamina propria. In contrast, low grade papillary tumors demonstrate densely arranged but normal-shaped small cells extending outward from fibrovascular cores. High grade tumors show markedly irregular architecture and cellular pleomorphism. CONCLUSIONS: We report the first study to our knowledge of in vivo confocal laser endomicroscopy in the urinary tract. Marked differences among normal urothelium, low grade tumors and high grade tumors were visualized. Pending further clinical investigation and technological improvement, confocal laser endomicroscopy may become a useful adjunct to conventional cystoscopy. PMID- 19683271 TI - Editorial comment. PMID- 19683272 TI - Longer wait times increase overall mortality in patients with bladder cancer. AB - PURPOSE: We used population level data to determine the impact of extended wait times on the survival of patients who underwent radical cystectomy for bladder cancer. MATERIALS AND METHODS: We identified 2,535 patients who underwent cystectomy for bladder cancer in Ontario, Canada between 1992 and 2004 using administrative databases. A Cox proportional hazards model accounting for patient, pathological and health services variables that could affect wait times was created to assess the impact of wait time on survival. The tumor stage specific impact of waiting for cystectomy was also assessed. Cox regression analysis that modeled wait time using cubic splines was used to determine a maximum wait time within which optimal care can be provided. RESULTS: Median wait time from transurethral bladder resection to cystectomy was 50 days. Unadjusted and adjusted analyses demonstrated that prolonged wait times were significantly associated with a lower overall survival rate. The relative hazard of death with increasing wait times appeared greater for low stage vs high stage cancers. The cubic splines regression analysis revealed that the risk of death began to increase after 40 days. CONCLUSIONS: Treatment delay between transurethral bladder tumor resection and radical cystectomy resulted in worse overall survival. The effect of wait time was greatest in lower stage lesions. The suggested maximum wait time from transurethral bladder tumor resection to cystectomy was 40 days. Further studies assessing disease-free survival are required to corroborate these findings. PMID- 19683273 TI - Editorial comment. PMID- 19683274 TI - Location, extent and number of positive surgical margins do not improve accuracy of predicting prostate cancer recurrence after radical prostatectomy. AB - PURPOSE: Positive surgical margins increase the risk of biochemical recurrence after radical prostatectomy by 2 to 4-fold. The risk of biochemical recurrence may be influenced by the anatomical location and extent of positive surgical margins. In a multicenter study we analyzed the predictive usefulness of several subclassifications of positive surgical margins. MATERIALS AND METHODS: The clinical information and followup data of 7,160 patients treated with radical prostatectomy alone at 1 of 3 institutions between 1995 and 2006 were modeled using Cox proportional hazards regression analysis for biochemical recurrence. Positive surgical margins were analyzed as solitary vs multiple, focal vs extensive and apical location vs other. The usefulness of these subclassifications was assessed by the improvement in predictive accuracy of nomograms containing these parameters compared to one in which the surgical margin was modeled simply as positive vs negative. RESULTS: The 7-year progression-free probability was 60% in patients with positive surgical margins. A positive surgical margin was significantly associated with biochemical recurrence (HR 2.3, p <0.001) after adjusting for age, prostate specific antigen, pathological Gleason score, pathological stage and year of surgery. An increased risk of biochemical recurrence was associated with multiple vs solitary positive surgical margins (adjusted HR 1.4, p = 0.002) and extensive vs focal positive surgical margins (adjusted HR 1.3, p = 0.004) on multivariable analysis. However, neither parameter improved the predictive accuracy of a nomogram compared to one in which surgical margin status was modeled as positive vs negative (concordance index 0.851 vs 0.850 vs 0.850). CONCLUSIONS: The number and extent of positive surgical margin significantly influence the risk of biochemical recurrence after radical prostatectomy. However, the empirical prognostic usefulness of subclassifications of positive surgical margins is limited. PMID- 19683275 TI - Initial experience with robot assisted partial nephrectomy for multiple renal masses. AB - PURPOSE: We evaluated the feasibility of performing robot assisted partial nephrectomy in patients with multiple renal masses and examined the results of our initial experiences. MATERIALS AND METHODS: We reviewed the records of 10 patients with multiple renal masses who underwent attempted robot assisted partial nephrectomy within the last 2 years. Demographic information, and intraoperative, perioperative and renal function outcome data on these patients were reviewed. RESULTS: A total of 24 tumors in 9 patients were removed with robot assistance. There was 1 open conversion with successful completion of partial nephrectomy. Of the patients 70% had a known hereditary renal cancer syndrome and the remainder had multifocal disease with unknown germline genetic alterations. Frozen section from the tumor bed evaluated in 5 of 10 cases was negative. One patient experienced urinary leak postoperatively, which resolved by postoperative day 9 without intervention. Of the 24 robotically resected masses 22 were malignant. Our most recent 3 patients underwent successful partial nephrectomy without hilar clamping, obviating the need for warm ischemia. Overall renal function was unchanged at most recent followup with a minimal decrease in operated kidney differential function. CONCLUSIONS: Robot assisted partial nephrectomy for multiple renal masses was feasible in our early experience. Patient selection is paramount for successful minimally invasive surgery. Robot assisted partial nephrectomy without hilar clamping, especially in the hereditary patient population in which repeat ipsilateral partial nephrectomy may be anticipated, appears promising but requires further evaluation. PMID- 19683276 TI - How bad are positive margins after radical prostatectomy and how are they best managed? PMID- 19683278 TI - Combined thermo-chemotherapy for recurrent bladder cancer after bacillus Calmette Guerin. AB - PURPOSE: Despite an initial adequate response many patients with nonmuscle invasive urothelial cell carcinoma of the bladder eventually have recurrence after intravesical bacillus Calmette-Guerin treatments. We evaluated the efficacy of combined bladder wall hyperthermia and intravesical mitomycin C instillation (thermo-chemotherapy) in cases of recurrence after bacillus Calmette-Guerin. MATERIALS AND METHODS: A total of 111 patients with recurrent papillary nonmuscle invasive urothelial cell carcinoma of the bladder after previous bacillus Calmette-Guerin treatment underwent complete bladder tumor resection and were referred for prophylactic adjuvant treatment with thermo-chemotherapy. Treatment was received on an outpatient basis weekly for 6 weeks, followed by 6 maintenance sessions at 4 to 6-week intervals. Each treatment included 2, 30-minute cycles of 20 mg mitomycin C and bladder wall hyperthermia to 42C +/- 2C. Cystoscopy and urine cytology were performed after the completion of induction treatment and every 3 months thereafter. RESULTS: The Kaplan-Meier estimated disease-free survival rate was 85% and 56% after 1 and 2 years, respectively. No maintenance treatment was associated with decreased efficacy, that is the recurrence rate was 61% at 2 years vs 39% in those with maintenance treatments (p = 0.01). The progression rate was 3%. CONCLUSIONS: Thermo-chemotherapy may be effective for papillary nonmuscle invasive urothelial cell carcinoma of the bladder that recurs after BCG treatment without increasing the risk of tumor progression. Maintenance therapy is important and improves the outcome. PMID- 19683279 TI - Editorial comment. PMID- 19683280 TI - Tertiary Gleason patterns and biochemical recurrence after prostatectomy: proposal for a modified Gleason scoring system. AB - PURPOSE: We investigated the relationship between the tertiary Gleason component in radical prostatectomy specimens and biochemical recurrence in what is to our knowledge the largest single institution cohort to date. MATERIALS AND METHODS: We evaluated data on 3,230 men who underwent radical prostatectomy at our institution from 2000 to 2005. Tertiary Gleason component was defined as Gleason grade pattern 4 or greater for Gleason score 6 and Gleason grade pattern 5 for Gleason score 7 or 8. RESULTS: Biochemical recurrence curves for cancer with tertiary Gleason component were intermediate between those of cancer without a tertiary Gleason component in the same Gleason score category and cancer in the next higher Gleason score category. The only exception was that Gleason score 4 + 3 = 7 with a tertiary Gleason component behaved like Gleason score 8. The tertiary Gleason component independently predicted recurrence when factoring in radical prostatectomy Gleason score, radical prostatectomy stage and prostate specific antigen (HR 1.45, p = 0.029). Furthermore, the magnitude of the tertiary Gleason component effect on recurrence did not differ by Gleason score category (p = 0.593). CONCLUSIONS: Although the tertiary Gleason component is frequently included in pathology reports, it is routinely omitted in other situations, such as predictive nomograms, research studies and patient counseling. The current study adds to a growing body of evidence highlighting the importance of the tertiary Gleason component in radical prostatectomy specimens. Accordingly consideration should be given to a modified radical prostatectomy Gleason scoring system that incorporates tertiary Gleason component in intuitive fashion, including Gleason score 6, 6.5 (Gleason score 6 with tertiary Gleason component), 7 (Gleason score 3 + 4 = 7), 7.25 (Gleason score 3 + 4 = 7 with tertiary Gleason component), 7.5 (Gleason score 4 + 3), 8 (Gleason score 4 + 3 with tertiary Gleason component or Gleason score 8), 8.5 (Gleason score 8 with tertiary Gleason component), 9 (Gleason score 4 + 5 or 5 + 4) and 10. PMID- 19683282 TI - PIC cystography: a selective approach to the diagnosis of vesicoureteral reflux. PMID- 19683281 TI - Tumor size is a determinant of the rate of stage T1 renal cell cancer synchronous metastasis. AB - PURPOSE: A recent multi-institutional analysis of 995 patients treated for renal cell cancer questioned the relationship between tumor size and the synchronous metastasis rate. We revisited the hypothesis that metastatic potential is unrelated to tumor size. MATERIALS AND METHODS: We tested the relationship between tumor size and synchronous metastasis in 22,204 patients with T1a and T1b renal cell cancer diagnosed and/or treated with nephrectomy for clear cell, papillary or chromophobe histological subtypes in 1 of 9 Surveillance, Epidemiology and End Results registries between 1988 and 2004. RESULTS: In the study population the synchronous metastasis rate was 9.6%, including 5.6% vs 14.2% for T1a vs T1b. Stratification by 1 cm tumor size intervals revealed that the rate increased with increasing tumor size, that is 4.8% at 1.0 cm or less, 4.2% at 1.1 to 2.0 cm, 4.9% at 2.1 to 3.0 cm, 7.1% at 3.1 to 4.0 cm, 12.1% at 4.1 to 5.0 cm, 13.3% at 5.1 to 6.0 cm and 18.4% 6.1 to 7.0 cm (chi-square trend p <0.001). Cubic spline analysis showed that tumor size was virtually linearly related to the synchronous metastasis rate. Stratification by histological subtype in patients treated with nephrectomy revealed that clear cell renal cell cancer was most frequently associated with synchronous metastasis. Finally, tumor size was an independent predictor of synchronous metastasis in multivariate regression models adjusted for age, gender, histological subtype and year of diagnosis quartiles. CONCLUSIONS: Our study confirms that tumor size is an important determinant of the likelihood of synchronous metastasis in patients with T1a and T1b renal cell cancer. The synchronous metastasis rate directly increases with increasing tumor size. Even patients with small renal masses are at risk for synchronous metastasis and patients with clear cell renal cell cancer are at highest risk. PMID- 19683283 TI - Is it time to revise our evaluation of hematuria? PMID- 19683284 TI - Fibrin clot inhibitor medication and efficacy of bacillus Calmette-Guerin for bladder urothelial cancer. AB - PURPOSE: Studies suggest that the antitumor effect of bacillus Calmette-Guerin depends on bacillus Calmette-Guerin attachment to fibronectin at fibrin clot formation sites and medications that impact fibrin clot formation may modify bacillus activity. We evaluated the impact of fibrin clot inhibitors on the clinical efficacy of bacillus Calmette-Guerin. MATERIALS AND METHODS: We reviewed the records of 907 consecutive patients treated with bacillus Calmette-Guerin between 1990 and 2006. Time to disease recurrence and progression to surgery were compared in patients who did and did not receive fibrin clot inhibitors by Kaplan Meier methods and multivariate Cox regression models. RESULTS: Overall 221 patients (24%) received at least 1 fibrin clot inhibitor, including 170, 34 and 52 on aspirin, clopidogrel and warfarin, respectively. Patients on warfarin had shorter time to progression than patients not on warfarin (median 2.1 vs 9.0 years, p <0.005). Patients on aspirin had a significantly improved 5-year probability of freedom from surgery (66% vs 56%, p = 0.029). On multivariate analysis warfarin was associated with an increased risk of progression to surgery (HR 1.89, 95% CI 1.31, 2.74, p = 0.0007), while aspirin was associated with a decreased risk (HR 0.71, 95% CI 0.52, 0.96, p = 0.024). Warfarin alone was associated with an increased risk of tumor recurrence (HR 1.39, 95% CI 1.00, 1.94, p = 0.047). CONCLUSIONS: These data suggest that the risks of recurrence and progression to surgery after bacillus Calmette-Guerin are higher in patients on warfarin, while the risk of progression is lower in patients on aspirin. These findings may have important treatment implications in patients in whom bacillus Calmette-Guerin is contemplated. PMID- 19683285 TI - Editorial comment. PMID- 19683286 TI - Pathological effects of prostate cancer correlate with neuroendocrine differentiation and PTEN expression after bicalutamide monotherapy. AB - PURPOSE: Androgen deprivation therapy is the primary treatment for advanced prostate cancer but many patients eventually experience progression to hormone refractory status. Understanding the molecular changes after androgen deprivation therapy would help evaluate the efficacy or failure of second line therapies. Therefore, we analyzed the expression of the tumor suppressor phosphatase and tensin homologue deleted on chromosome 10 (PTEN), the human epidermal receptor-2 and neuroendocrine differentiation after bicalutamide monotherapy, which is emerging as an alternative treatment for locally advanced prostate cancer. MATERIALS AND METHODS: Molecular arrangements were evaluated in 107 radical prostatectomy specimens from patients given 150 mg bicalutamide before surgery. Pathological regressive changes, and the correlation of postoperative biochemical failure with the extent of molecular arrangements and pathological effects were analyzed. RESULTS: Patients with minimal regression effects after bicalutamide therapy had advanced pathological stage disease, and tended to have positive chromogranin A expression and PTEN inactivation. Only 4 (3.7%) prostatectomy specimens showed human epidermal receptor-2 immunostaining. The probability of positive chromogranin A expression in the PTEN inactivation group was 2.5-fold (OR 2.5, 95% CI 1.1-5.6, p = 0.023) higher than in the nonPTEN inactivation group. Cox regression analysis revealed that seminal vesicle invasion, PTEN/chromogranin A expression and lymph node invasion were significant variables for time to biochemical recurrence. CONCLUSIONS: PTEN inactivation and neuroendocrine differentiation were related to refractoriness to bicalutamide therapy. These results support the hypothesis that neuroendocrine differentiation is caused by activation of the serine threonine kinase Akt pathway, which results from PTEN inactivation. PMID- 19683287 TI - Hyaluronic acid and HYAL-1 in prostate biopsy specimens: predictors of biochemical recurrence. AB - PURPOSE: Molecular markers could aid prostate specific antigen, biopsy Gleason sum and clinical stage to provide accurate information on prostate cancer progression. HYAL-1 hyaluronidase and hyaluronic acid staining in prostatectomy specimens predicts biochemical recurrence. We examined whether hyaluronic acid and HYAL-1 staining in biopsy specimens predicts biochemical recurrence and correlates with staining in matched prostatectomy specimens. MATERIALS AND METHODS: Biopsy and prostatectomy specimens were obtained from 61 patients with clinically localized prostate cancer from multiple centers, including 23 with (group 1) and 38 without (group 2) biochemical recurrence. Mean followup was 103.1 months. Biotinylated hyaluronic acid binding protein and anti-HYAL-1 antibody were used for hyaluronic acid and HYAL-1 staining, respectively. Staining was graded between 0 and 300 depending on staining intensity and area. RESULTS: HYAL-1 and hyaluronic acid were expressed in tumor cells and stroma, respectively. In biopsy specimens HYAL-1 and hyaluronic acid expression was higher in group 1 than in group 2 (203.9 and 182.1 vs 48.8 and 87.0, respectively, p <0.0001). On univariate analysis hyaluronic acid, HYAL-1, biopsy Gleason and prostate specific antigen significantly predicted biochemical recurrence (p <0.001). On multivariate analysis only HYAL-1 staining independently predicted recurrence with an accuracy of 81.8% (p <0.001). In prostatectomy specimens only HYAL-1 staining correlated with staining in biopsy specimens (Spearman rho = 0.72, p = 0.0002) and predicted biochemical recurrence. CONCLUSIONS: To our knowledge this is the first report that HYAL-1 staining in biopsy specimens is an independent predictor of biochemical recurrence. This may be useful when selecting treatment. PMID- 19683288 TI - Editorial comment. PMID- 19683289 TI - Ketoconazole and prednisone to prevent recurrent ischemic priapism. AB - PURPOSE: To our knowledge no standard therapy exists for the prevention of recurrent ischemic priapism. We used ketoconazole and prednisone with dosing titrated according to serum testosterone levels to suppress sleep related erections in an attempt to prevent recurrent episodes. MATERIALS AND METHODS: Eight patients with recurrent ischemic priapism were treated with ketoconazole and prednisone. Two patients had sickle cell anemia and 6 had idiopathic recurrent ischemic priapism. Testosterone was measured on initial presentation, and ketoconazole and prednisone dosing was titrated to approximately 200 ng/dl testosterone and based on the presence or absence of recurrent ischemic priapism episodes. The International Index of Erectile Function-5 questionnaire was administered to evaluate for erectile dysfunction. Patients were seen monthly and therapy was withdrawn after 6 months. RESULTS: Mean testosterone before and after treatment was 468 and 275 ng/dl, respectively. Mean followup was approximately 1.5 years. One patient had 2 recurrent ischemic priapism episodes while on ketoconazole and prednisone treatment. Another patient experienced an increase in testosterone from 361 to 432 ng/dl after initiation of therapy, and 3 recurrent ischemic priapism episodes requiring emergency corporal irrigation. After dose titration testosterone was 184 ng/dl and the patient has had no subsequent episodes. Mean International Index of Erectile Function-5 score was 24.8 points. There were no recurrent ischemic priapism episodes after withdrawal of ketoconazole and prednisone, and no reported symptoms of hypogonadism. CONCLUSIONS: Ketoconazole and prednisone therapy was well tolerated in these 8 patients with recurrent ischemic priapism, and with testosterone monitoring and dose titration it was successful in preventing recurrent episodes while preserving sexual function. PMID- 19683290 TI - Intravesical liposome versus oral pentosan polysulfate for interstitial cystitis/painful bladder syndrome. AB - PURPOSE: We evaluated the safety and efficacy of intravesical liposomes, a mucosal protective agent, compared to oral pentosan polysulfate sodium for interstitial cystitis/painful bladder syndrome. MATERIALS AND METHODS: We performed a prospective longitudinal study of the effect of 2 independent treatments (intravesical liposomes and oral pentosan polysulfate sodium) in patients with interstitial cystitis/painful bladder syndrome. Ten possible responses (or measures) to treatment were monitored at 3 time points, including baseline, and weeks 4 and 8. A total of 24 patients with interstitial cystitis/painful bladder syndrome were evaluated in a 1:1 ratio to intravesical liposomes (80 mg/40 cc distilled water) once weekly or to oral pentosan polysulfate sodium (100 mg) 3 times daily for 4 weeks each. RESULTS: No patient had urinary incontinence, retention or infection due to liposome instillation. There were no unanticipated adverse events and no significant worsening of symptoms during followup. Statistically significant decreases in urinary frequency and nocturia were observed in each treatment group. Statistically significant decreases in pain, urgency and the O'Leary-Sant symptom score were observed in the liposome group. Decreased urgency in the liposome group had the most profound effect of the ordinal measures. CONCLUSIONS: Each glycosaminoglycan directed treatment seemed beneficial. Liposome intravesical instillation is safe for interstitial cystitis/painful bladder syndrome with potential improvement after 1 course of therapy for up to 8 weeks. Intravesical liposomes achieved efficacy similar to that of oral pentosan polysulfate sodium. Further large-scale placebo controlled studies are needed. Intravesical liposomes appear to be a promising new treatment for interstitial cystitis/painful bladder syndrome. PMID- 19683291 TI - Editorial comment. PMID- 19683293 TI - Editorial comment. PMID- 19683294 TI - Predictors of surgical approach to repair pelvic fracture urethral distraction defects. AB - PURPOSE: We identified preoperative factors predictive of the appropriate surgical approach to anastomotic repair of pelvic fracture urethral distraction defects. MATERIALS AND METHODS: We reviewed the medical records and imaging studies of 121 patients who had undergone anastomotic repair of a pelvic fracture urethral distraction defect. The review was focused on 10 preoperative clinicoradiological variables that may influence or predict the surgical repair. The patients were categorized as having undergone a simple perineal operation (78 patients, group 1), or an elaborated perineal or a combined perineo-abdominal procedure (43 patients, group 2). Univariate and multivariate analyses were used to identify preoperative parameters predictive of the type of anastomotic repair. In addition, ROC analysis was used to assess prediction results of the multivariate analysis. RESULTS: On univariate analysis 5 parameters were significant predictors of the type of repair, while on multivariate analysis only 3 parameters remained strong and independent predictors including the gapometry/urethrometry index, urethral gap length and prostatic displacement. The gapometry/urethrometry index was a proxy for all other parameters. At a cutoff index of 0.35 the appropriate surgical repair was predicted with 91% specificity and a 95% positive predictive value. When ROC analysis was performed the AUC was 0.979. CONCLUSIONS: The type of anastomotic repair of pelvic fracture urethral distraction defect can be predicted by 3 preoperative factors, namely the gapometry/urethrometry index, urethral gap length and prostatic displacement. The gapometry/urethrometry index has the highest predictive accuracy and is a proxy for all other factors. An index less than 0.35 indicates a simple perineal operation and an index greater than 0.35 indicates an elaborated perineal or a transpubic procedure. PMID- 19683295 TI - Sacral nerve stimulation for refractory overactive bladder in the elderly population. AB - PURPOSE: We determined the long-term outcome of sacral nerve stimulation for refractory overactive bladder in the elderly population. MATERIALS AND METHODS: We performed a prospective longitudinal study to better characterize the outcome of sacral nerve stimulation in female patients 70 years old or older with refractory overactive bladder. Demographic and perioperative data were recorded. Patients were followed postoperatively for evidence of successful stage conversion, device durability and efficacy, and postoperative complications. Patients were retrospectively compared to a cohort of female patients younger than 70 years with refractory overactive bladder. Statistical analysis was performed. RESULTS: Between July 2001 and February 2008, 19 elderly female patients with refractory overactive bladder underwent stage 1 lead placement. Of the patients 17 (90%) who reported greater than 50% improvement in symptoms based on a 1-week followup voiding log underwent implantable pulse generator placement. No intraoperative or immediate postoperative complications were noted. At a mean followup of 48.5 months 11 patients (65%) had a functional implantable pulse generator with greater than 50% objective improvement over baseline. Compared to matched patients younger than 70 years elderly patients had a similar conversion rate and adverse events but were significantly more likely to undergo device removal (p = 0.018). CONCLUSIONS: Based on our experience elderly patients have a high conversion rate, few adverse events, and a high level of device efficacy and durability with sacral nerve stimulation. Although more mature multicenter data are needed, it appears that sacral nerve stimulation in geriatric patients is safe and efficacious, and should be judiciously offered to those with refractory voiding symptoms. PMID- 19683296 TI - Novel electromagnetic lithotriptor for upper tract stones with and without a ureteral stent. AB - PURPOSE: We compared the treatment efficacy and safety of the novel Lithoskop electromagnetic extracorporeal shock wave lithotriptor for upper urinary tract stones with and without prior ureteral stent placement. MATERIALS AND METHODS: A total of 665 consecutive patients harboring single renal or ureteral stones underwent shock wave lithotripsy between August 2006 and July 2008. In 75 and 46 patients with renal and ureteral stones, respectively, stents were placed before the first shock wave lithotripsy session. Treatment outcome was assessed in relation to stent placement. All data were derived from a prospectively maintained database. RESULTS: The mean size of nonstented vs stented renal and ureteral stones was 8.6 vs 12.5 mm (p <0.0001) and 7.1 vs 7.3 mm (p = 0.6), respectively. The stone-free rate in nonstented vs stented renal and ureteral stone cases was 76.3% vs 77.3% and 91.4% vs 93.5%, respectively (each p >0.99). The total energy applied per stone was 110 +/- 83 vs 150 +/- 89 J (p <0.0001) and 183 +/- 131 vs 209 +/- 125 J (p = 0.1), respectively. Auxiliary measures were required after shock wave lithotripsy for renal and ureteral stones in 5.4% and 10.8% of nonstented, and in 1.3% and 4.3% of stented cases, respectively. No complications were detected in stented renal and ureteral stone cases compared to 2.9% and 6.9% in nonstented cases, respectively. CONCLUSIONS: A high success rate and a low complication rate were achieved in renal and ureteral stone cases with and without prior ureteral stent placement. Total energy needed to achieve a stone-free state did not differ between stented and nonstented ureteral cases, suggesting the absence of a significant influence of the stent. Overall stents decreased complications necessitating hospitalization and auxiliary invasive measures. PMID- 19683298 TI - Botulinum toxin A for overactive bladder and detrusor muscle overactivity in patients with Parkinson's disease and multiple system atrophy. AB - PURPOSE: Urinary disturbances are common in patients with Parkinson's disease and multiple system atrophy. We investigated the effectiveness and safety of botulinum toxin type A injected into the detrusor muscle in patients with Parkinson's disease and multiple system atrophy who had refractory overactive bladder symptoms and detrusor overactivity. MATERIALS AND METHODS: All participants underwent clinical and urodynamic assessment, and completed a quality of life questionnaire before botulinum toxin type A treatment, and 1 and 3 months thereafter. Four patients with Parkinson's disease and 2 with multiple system atrophy were enrolled in the study. All patients received 200 U botulinum toxin type A injected into the detrusor muscle at 20 sites under cystoscopic guidance at a single session on an inpatient basis. Outcome measures were clinical assessment (a voiding diary including daytime and nighttime urinary frequency, and episodes of urgency and urge urinary incontinence), urodynamic assessment (including first volume and maximum pressure of uninhibited detrusor contractions, and maximum cystometric capacity) and pressure flow studies. RESULTS: One and 3 months after botulinum toxin type A injection all patients reported that daytime and nighttime urinary frequency had decreased and quality of life scores improved. No patients had further episodes of urgency and urge urinary incontinence during the 5-month followup. Urodynamics showed improvement in all urinary function variables tested. No systemic side effects were recorded during or after treatment. In all patients post-void urinary residual volume increased and intermittent catheterization was required only in those with multiple system atrophy. CONCLUSIONS: The new beneficial effect that we report in a small study sample encourages larger trials to confirm botulinum toxin type A injection into the detrusor muscle as an effective and safe treatment for refractory overactive bladder symptoms and detrusor overactivity related to Parkinson's disease and multiple system atrophy. PMID- 19683299 TI - New insights into the role of interleukin-6 in human prostate cancer. PMID- 19683300 TI - Incidence of recurrent frank hematuria and urological cancers: prospective 6.9 years of followup. AB - PURPOSE: The majority of patients presenting with frank hematuria have no diagnosis. There is a paucity of literature on the recurrence of frank hematuria and the incidence of urological cancers in these patients, and this study addresses both issues. MATERIALS AND METHODS: We performed a prospective cohort study of 578 consecutive patients referred with frank hematuria between 1999 and 2001 who underwent full investigations with a mean followup of 6.9 years. The primary outcome measure was the probability of frank hematuria recurrence after the initial negative investigations and the incidence of urological cancers in these patients. RESULTS: Diagnosis was made in 206 (35.6%) patients at initial presentation. Diagnosis was not made at initial presentation in the remaining 372 (64.4%) patients, of whom 81 died without a diagnosis during followup (32 within 2 years of presentation). A total of 81 patients (21.8%) with no diagnosis died during the followup period (32 within 2 years of the investigations). A questionnaire was mailed to the remaining 291 patients and 202 (69.4%) responded. Of the responders 41 (20.3%) reported frank hematuria recurrence (single episode in 10 and multiple episodes in 31). A significant urological diagnosis was made upon repeat evaluation in 21 (10.4%) patients which included urological malignancy in 4 (2%). CONCLUSIONS: Approximately 80% of cases cleared by initial investigation remained clear and 9.8% with frank hematuria recurrence were diagnosed with a urological malignancy. Frank hematuria recurrence requires vigilance and repeat investigations as appropriate. PMID- 19683301 TI - Editorial comment. PMID- 19683302 TI - Do oral antimuscarinic drugs carry an increased risk of acute urinary retention? AB - PURPOSE: We determined whether men treated with oral antimuscarinics are at increased risk for acute urinary retention. MATERIALS AND METHODS: In this population based, retrospective cohort study using a nested case-control design we analyzed data from a large primary care database containing patient information entered by general practitioners in the United Kingdom. Our study cohort comprised men 20 to 84 years old. Cases of acute urinary retention were identified by reviewing diagnostic codes and were confirmed in a random sample through questionnaires sent to the treating physician. RESULTS: The overall incidence of acute urinary retention in the study cohort (1,844) was 1.0 per 1,000 person-years, with the incidence rate increasing with age. The first 30 days (early treatment) of antimuscarinic use was associated with a relative risk of acute urinary retention of 8.3 (95% CI 4.8-14.2) and with longer term use (more than 30 days) the relative risk was 2.0 (95% CI 1.2-3.1). The relative risk of acute urinary retention was similar for low/medium and high antimuscarinic doses (relative risk 2.8 vs 3.0, 95% CI 2.1-3.8 and 1.3-6.8, respectively). The relative risk of acute urinary retention was highest during early treatment for a urogenital indication (relative risk 14.2, 95% CI 6.8-29.6). The risk of acute urinary retention was not increased when antimuscarinics were used as antispasmodics or for drug induced parkinsonism. CONCLUSIONS: Men prescribed antimuscarinics, particularly for a urogenital condition, should be closely monitored during the first 30 days of treatment for signs or symptoms of urinary retention. PMID- 19683303 TI - A randomized, double-blind trial to compare shock wave frequencies of 60 and 120 shocks per minute for upper ureteral stones. AB - PURPOSE: Recent evidence demonstrates that decreasing shock wave frequency from the previous standard of 120 to 60 shocks per minute results in improved fragmentation of stones located within the renal collecting system. We report the first randomized trial to our knowledge to examine the effect of a slower shock wave frequency for shock wave lithotripsy on stones located in the proximal ureter. MATERIALS AND METHODS: A total of 163 patients with a previously untreated radiopaque calculus in the upper ureter measuring at least 5 mm underwent stratified block randomization according to stone size, and shock wave lithotripsy at 60 or 120 shocks per minute. Stone-free status at 3 months was confirmed with noncontrast computerized tomography or a plain abdominal x-ray and ultrasound study. RESULTS: Of the patients 77 were randomized to 60 shocks per minute and 86 were randomized to 120 shocks per minute. The groups were similar in gender, age, body mass index and initial stone area. At 3 months the 60 shocks per minute group had a higher overall stone-free rate (64.9% vs 48.8%, p = 0.039). Significantly fewer shocks were administered to patients treated at 60 shocks per minute (mean 2,680 vs 2,940, p <0.001). However, mean treatment times were longer (44.3 vs 24.5 minutes, p <0.001). Patients treated with 60 shocks per minute required fewer auxiliary procedures (29.9% vs 45.4%) (p = 0.031). CONCLUSIONS: Decreasing the rate of shock wave administration from 120 to 60 shocks per minute results in improved stone-free rates. A slower treatment rate of proximal ureteral stones reduces the need for additional shock wave lithotripsy or more invasive treatments to render patients stone-free, without any increase in morbidity, and with an acceptable increase in treatment time. PMID- 19683304 TI - Long-term outcome of epididymectomy for the management of chronic epididymal pain. AB - PURPOSE: We evaluated the long-term outcomes of patients who underwent epididymectomy for the treatment of chronic epididymal pain. MATERIALS AND METHODS: All 72 patients who underwent epididymectomy at our institution between 1994 and 2007 were invited to participate in the study. Patients were mailed questionnaires covering various aspects of the treatment. Questions regarding pain were rated on a scale between 0 and 10 (0--no pain, 10--severe pain). Patients who did not return the questionnaires were followed up by telephone and the medical case notes of all respondents were reviewed. Statistical analysis was performed using the Wilcoxon signed-rank and Fisher's exact tests with p <0.05 considered statistically significant. RESULTS: A total of 53 patients participated (74% response rate) and mean followup was 7.4 years. Of these patients 45 (84.9%) underwent epididymectomy for post-vasectomy pain and the remainder (8 of 53, 15.1%) had the procedure for various nonvasectomy reasons. There were significant improvements in pain score in the post-vasectomy (mean 7.3 preoperative to 2.4 postoperative, p <0.001) and nonvasectomy (mean 7 preoperative to 2.8 postoperative, p = 0.002) groups. Of the patients in the post vasectomy group 93.3% (42 of 45) had less or no pain postoperatively compared to 75% (6 of 8) in the nonvasectomy group. The satisfaction rate with epididymectomy was also higher in the post-vasectomy (42 of 45, 93.3%) compared to the nonvasectomy (5 of 8, 62.5%) group (p = 0.038). CONCLUSIONS: With high patient satisfaction and a favorable long-term outcome epididymectomy appears to be an effective treatment option particularly for post-vasectomy chronic epididymal pain. PMID- 19683305 TI - Prostate volume changes over time: results from the Baltimore Longitudinal Study of Aging. AB - PURPOSE: According to a 1944 publication by Swyer benign prostatic hyperplasia develops in some men after age 45 with further prostatic growth whereas in other men prostate size remains stable or decreases with advancing age. Although there is an abundance of literature describing prostatic enlargement in association with benign prostatic hyperplasia, less is known about the phenomenon of prostate atrophy. MATERIALS AND METHODS: In the Baltimore Longitudinal Study of Aging serial pelvic magnetic resonance imaging was performed in men without prostate cancer beginning in 1993. From this population we retrospectively identified 278 men with 2 or more magnetic resonance imaging determined prostate volume measurements to examine differential growth rates in a cohort of community men over time. RESULTS: Median age was 58 years and median prostate size was 28 cc at study entry. At a median followup of 4.3 years prostate size increased in 61.9% and remained stable or decreased in 38.1% of men. The median rate of volume change was 0.6 cc per year (range -9.9 to 62.1), corresponding to a median growth rate of 2.5% per year (range -29.2 to 176.4%). During followup 64.6% of men with an initial prostate size less than 40 cc had prostate growth compared to only 50.9% of men with an initial prostate size of 40 cc or greater. CONCLUSIONS: These results suggest that changes in prostate size are highly variable among aging men. Although benign prostatic hyperplasia is common, a considerable proportion of aging men have a stable or decreasing prostate size. Further research is needed to identify the underlying mechanism for such differences in prostate growth. PMID- 19683306 TI - Menopausal hormone therapy--why do different studies have different results for the same outcome? PMID- 19683307 TI - Should we centralize referrals for repair of urethral stricture? PMID- 19683308 TI - Vesicoureteral reflux nomograms--can we use these in the clinic? PMID- 19683309 TI - Stricture recurrence after urethroplasty: a systematic review. AB - PURPOSE: Urethroplasty remains the gold standard for the management of urethral stricture disease with acceptable long-term success. However, the standard by which stricture recurrence is defined and evaluated after urethral reconstruction remains widely variable. We conducted a systematic review of the urological literature to determine how stricture recurrence is defined and evaluated. MATERIALS AND METHODS: A systematic review was conducted on all contemporary urethroplasty articles published between 2000 and 2008. Using the term "urethroplasty" 302 articles were identified and evaluated. A total of 86 articles were included in the analysis. RESULTS: The overall recurrence rate for all reconstructive procedures was 15.6%, which remained stable between 2000 and 2008. Stricture recurrence was determined by a mean of 3 (range 1 to 8) different diagnostic tests. The most common primary diagnostic tests for recurrence were uroflowmetry (56% of articles) and retrograde urethrography (51%). Cystourethroscopy was used as a primary screen to identify stricture recurrence in 25% of articles, and as a secondary procedure in another 21%. Recurrence was defined as the need for an additional surgical procedure or dilation in 75% and 52% of articles, respectively. CONCLUSIONS: The methods used to determine stricture recurrence after urethroplasty remain widely variable. The use of a standardized surveillance protocol to define stricture recurrence after urethral reconstruction may allow more effective comparison of urethroplasty outcomes across institutions. PMID- 19683310 TI - Extensive biopsies and transurethral prostate resection in men with previous negative biopsies and high or increasing prostate specific antigen. AB - PURPOSE: We determined the diagnostic role of an extensive biopsy protocol associated with transurethral prostate resection in patients with persistently increased or increasing prostate specific antigen without evidence of prostate cancer after 2 or more extended negative sets of biopsies. MATERIALS AND METHODS: A new set of 21-core biopsies was done in 113 patients under general anesthesia in association with transurethral prostate resection. Demographics, clinical and biological data, operative parameters, pathological results and followup were recorded prospectively. RESULTS: Extended biopsies provided an 18.6% detection rate and detected 77.8% of prostate cancers. Transurethral prostate resection significantly increased the detection rate by 28.5% for an overall 23.9% prostate cancer detection rate (p = 0.035). Most prostate cancer detected on chips and/or biopsy was clinically significant and 30% were scored as Gleason 7 or greater. Of prostatectomy specimens 19% showed pT3a-pT4 cancer with a median Gleason score of 7. In patients with no cancer mean prostate specific antigen 1 year after transurethral prostate resection was 4.5 ng/ml (range 0.3 to 16.3), which remained stable during followup. A third of these patients underwent repeat biopsy with a 16.7% prostate cancer detection rate. CONCLUSIONS: About a fourth of patients with at least 2 extended negative sets of prostate biopsies remain at risk for prostate cancer and most tumors missed on initial procedures are clinically significant. Repeat biopsy using general anesthesia detects three fourths of these prostate cancers. However, the diagnostic yield of transurethral prostate resection appears significant and may provide additional data of clinical importance in select, informed patients. PMID- 19683312 TI - Editorial comment. PMID- 19683313 TI - Alcohol consumption is associated with a decreased risk of benign prostatic hyperplasia. AB - PURPOSE: While some studies have indicated that alcohol consumption is associated with a decreased risk of benign prostatic hyperplasia, others have not. We evaluated associations of alcohol consumption with benign prostatic hyperplasia and male lower urinary tract symptoms. MATERIALS AND METHODS: We performed a meta analysis of published studies pertaining to alcohol intake, benign prostatic hyperplasia and lower urinary tract symptoms. We analyzed abstracted data with random effects models to obtain pooled odds ratios of adjusted effects estimates. RESULTS: A total of 19 studies (120,091 men) met selection criteria and of these studies 14 revealed a significantly decreased likelihood of benign prostatic hyperplasia or lower urinary tract symptoms with increased alcohol intake. Sixteen studies were eligible for pooled analyses, of which 12 used benign prostatic hyperplasia as the primary outcome. We stratified total alcohol intake by gm per day into 6 strata. Alcohol intake was associated with a significantly or marginally significantly decreased likelihood of benign prostatic hyperplasia in all 6 strata (p values 0.08, 0.01, <0.001, 0.02, 0.001 and <0.001, respectively). Compared to no alcohol intake, an alcohol intake of 36 gm daily or greater was associated with a 35% decreased likelihood of benign prostatic hyperplasia (OR 0.65, 95% CI 0.58-0.74, p <0.001). Of the 4 studies that used lower urinary tract symptoms as the primary outcome 3 demonstrated a significantly increased likelihood of lower urinary tract symptoms with alcohol consumption. CONCLUSIONS: Alcohol consumption is associated with a decreased likelihood of benign prostatic hyperplasia but not of lower urinary tract symptoms. Further studies are needed to determine the mechanisms by which alcohol modifies the risk of benign prostatic hyperplasia. PMID- 19683314 TI - Behavioural responses of the mangrove fiddler crabs (Uca annulipes and U. inversa) to urban sewage loadings: results of a mesocosm approach. AB - The study aimed at investigating the effects of sewage loadings on the behaviour of two fiddler crabs species maintained in a system of experimental mesocosms, built in a mangrove area in Tanzania and inundated with different seawater/sewage mixtures. Our results show that sewage loads led to a modification of the overall activity budget of the crab community as a result of increased hypertrophic conditions (high COD, increased chlorophyll-a concentrations). During their activity period, crabs inside contaminated mesocosms seemed to satisfy their feeding demand faster than those of the control cells, spending a significant longer time in other activities like courtship and territorial defence. Apart from being a good biological indication of ecosystem eutrophication, such a reduced foraging activity by fiddler crabs also depresses their sediment bioturbation activity, important factor for the health of mangrove systems, suggesting practical implications regarding the efficiency of mangrove-based wetlands for treatment of domestic sewage. PMID- 19683315 TI - [Vitamin B12 deficiency secondary to metformin therapy]. PMID- 19683316 TI - [Serological diagnosis of Q fever: a pending matter?]. PMID- 19683317 TI - [Erythema ab igne]. PMID- 19683318 TI - ICChI, a glycosylated chitinase from the latex of Ipomoea carnea. AB - A multi-functional enzyme ICChI with chitinase/lysozyme/exochitinase activity from the latex of Ipomoea carnea subsp. fistulosa was purified to homogeneity using ammonium sulphate precipitation, hydrophobic interaction and size exclusion chromatography. The enzyme is glycosylated (14-15%), has a molecular mass of 34.94 kDa (MALDI-TOF) and an isoelectric point of pH 5.3. The enzyme is stable in pH range 5.0-9.0, 80 degrees C and the optimal activity is observed at pH 6.0 and 60 degrees C. Using p-nitrophenyl-N-acetyl-beta-D-glucosaminide, the kinetic parameters K(m), V(max), K(cat) and specificity constant of the enzyme were calculated as 0.5mM, 2.5 x 10(-8)mol min(-1)microg enzyme(-1), 29.0 s(-1) and 58.0mM(-1)s(-1) respectively. The extinction coefficient was estimated as 20.56 M(-1)cm(-1). The protein contains eight tryptophan, 20 tyrosine and six cysteine residues forming three disulfide bridges. The polyclonal antibodies raised and immunodiffusion suggests that the antigenic determinants of ICChI are unique. The first fifteen N-terminal residues G-E-I-A-I-Y-W-G-Q-N-G-G-E-G-S exhibited considerable similarity to other known chitinases. Owing to these unique properties the reported enzyme would find applications in agricultural, pharmaceutical, biomedical and biotechnological fields. PMID- 19683319 TI - Absolute configuration of (+)-pinoresinol 4-O-[6''-O-galloyl]-beta-D glucopyranoside, macarangiosides E, and F isolated from the leaves of Macaranga tanarius. AB - A lignan glucoside, (+)-pinoresinol 4-O-[6''-O-galloyl]-beta-D-glucopyranoside (1), and two megastigmane glucosides, named macarangiosides E and F (2,3), together with 15 known compounds (4-18) were isolated from leaves of Macaranga tanarius (L.) Mull.-Arg. (Euphorbiaceae). Their structures were elucidated by spectroscopic and chemical analyses. In addition, the absolute stereochemistry of macarangiosides B and C isolated previously from the same plant was also determined for the first time. Compounds 1 and 2 were galloylated on glucose and possessed potent DPPH radical-scavenging activity. PMID- 19683320 TI - Gentisic acid conjugates of Medicago truncatula roots. AB - Three phenolic glycosides 5-O-{[5''-O-E-(4'''-O-threo-guaiacylglycerol)-feruloyl] beta-apiofuranosyl-(1-->2)-beta-xylopyranosyl} gentisic acid, 5-O-[(5''-O vanilloyl)-beta-apiofuranosyl-(1-->2)-beta-xylopyranosyl] gentisic acid and 1-O [E-(4'''-O-threo-guaiacylglycerol)-feruloyl]-3-O-beta-galacturonopyranosyl glycerol were isolated and identified from the roots of Medicago truncatula together with four known 5-O-beta-xylopyranosyl gentisic acid, vicenin-2, hovetrichoside C and pterosupin identified for the first time in this species. Structural elucidation was carried out on the basis of UV, mass, (1)H and (13)C NMR spectral data. PMID- 19683321 TI - Long-term immunogenicity and protection against Mycoplasma agalactiae induced by an oil adjuvant vaccine in sheep. AB - The long-term protective immunity of an inactivated mineral-oil adjuvanted Mycoplasma agalactiae vaccine was evaluated in sheep. The antigen suspension was emulsified with a mixture of three mineral oils (Montanide ISA-563, Marcol-52, Montane-80 at the ratio of 30%, 63%, and 7%, respectively). Twenty-two animals were divided in 2 groups (A and B) and immunised with two doses of the vaccine (group A, n=14) or used as unvaccinated control (group B, n=8). Five months after the second vaccination, seven animals of group A and four animals of group B were challenged by nasal route with M. agalactiae. The remaining seven vaccinated and four control animals were challenged intranasally eight months after vaccination. The vaccine was able to induce a full-protective immunity preventing the clinical signs of contagious agalactia and the infection by M. agalactiae in all groups of animals irrespective of the time of challenge after booster administration. PMID- 19683322 TI - A comparative study by age and gender of the pituitary adenoma and ACTH and alpha MSH secretion in dogs with pituitary-dependent hyperadrenocorticism. AB - Pituitary-dependent hyperadrenocorticism (PDH) is frequent in dogs. Little is known about its presentation in different age groups and its characteristics. Dividing the population under study (n=107) into three age groups we observed that 11.2% were young, 51.4% adults and 37.4% aged. Using magnetic resonance, pituitary tumours were intra-sellar (IS) in 30.8% and extra-sellar (ES) in 62.6% and the pars intermedia (PI) was affected in 6.5%. ES are predominant in females and IS in males (p<0.0001). In the adult-aged population, the ES and PI are predominant, while in the young, the IS predominate (p<0.0001). ACTH concentration was greater in the ES vs. IS (p<0.05). alpha-MSH did not present significant differences according to tumour size, showing a negative correlation (r=-0.47; p<0.01) vs. ACTH. Differences in adenoma size according to gender and their age-related frequency of apparition could be because of different origins of the corticotrophinoma. PMID- 19683323 TI - [Postictal psychosis]. AB - In epilepsy patients, psychotic states are related to a group of psychotic disorders with a specific phenomenology in which potential pathophysiological mechanisms are believed to be closely related to the epileptic disorder itself. Postictal psychosis is a very specific syndrome in relation to seizure activity: a clear temporal relationship exists between the psychotic state of sudden onset and a precipitating bout of complex partial or generalized seizures, with a characteristic lucid interval which lasts from two to 120h. The psychotic state may be related to the withdrawal of anticonvulsants, often in connection with video-EEG monitoring. The phenomenology of the psychotic state is often pleomorphic, with abnormal mood, paranoid delusions and hallucinations, with some clouding of consciousness or no evidence of impaired consciousness. The outcome is characterized by a remission of the psychotic symptoms over several days (mean: 1 week), with or without neuroleptic treatment. The majority of the patients suffer from complex partial seizures with frequent psychic auras that secondarily become generalized. In the majority of cases, prepsychotic EEG abnormalities persist during the psychosis. Frequent bitemporal foci are recorded on the EEG and MRI abnormalities (including mesial temporal sclerosis) are seen in more than half of the cases. The results of clinical, morphologic and metabolic available studies will be briefly discussed. PMID- 19683324 TI - Application of the combination index (CI)-isobologram equation to study the toxicological interactions of lipid regulators in two aquatic bioluminescent organisms. AB - Pharmaceuticals in the aquatic environment do not appear singly and usually occur as complex mixtures, whose combined effect may exhibit toxicity to the aquatic biota. We report an environmental application of the combination index (CI) isobologram equation, a method widely used in pharmacology to study drug interactions, to determine the nature of toxicological interactions of three fibrates toward two aquatic bioluminescent organisms, Vibrio fischeri and the self-luminescent cyanobacterial recombinant strain Anabaena CPB4337. The combination index-isobologram equation method allows computerized quantitation of synergism, additive effect and antagonism. In the Vibrio test, the fibrate combinations showed antagonism at low effect levels that turned into an additive effect or synergism at higher effect levels; by contrast, in the Anabaena test, the fibrate combinations showed a strong synergism at the lowest effect levels and a very strong antagonism at high effect levels. We also evaluated the nature of the interactions of the three fibrates with a real wastewater sample in the cyanobacterial test. We propose that the combination index-isobologram equation method can serve as a useful tool in ecotoxicological assessment. PMID- 19683325 TI - Behavior and fate of alkylphenols in surface water of the Jialu River, Henan Province, China. AB - The behavior and fate of alkylphenols (APs) were studied in surface water from the Jialu River, Henan Province, China. Located at its upper stream, Zhengzhou city is regarded as the major discharge source to this river with its annual effluents containing 726 kg for nonylphenol (NP) and 30.2 kg for octylphenol (OP). The concentrations of NP and OP in surface water ranged from 75.2 to 1520 ngL(-1) and from 20.9 to 63.2 ngL(-1), respectively. To assess the behavior of APs along the river, a mass balance equation based on chloride was adopted, due to its relative conservation. The results showed that dilution effect was prevailing in determining the APs concentrations in surface water along the river. The effect of potential biodegradation was also estimated with an assumption of the optimized biodegradation. The contributions of dilution and biodegradation to the decline of APs concentrations were 38.8%, 23.7% for NP and 57.8%, 24.3% for OP, respectively. The other contribution to the decline of APs concentrations along the river was considered as an integrated effect of adsorption and air-water exchange with the values of 37.5% for NP and 17.9% for OP. The decay half-lives of NP and OP from surface water bodies were 1.6 and 2.4d, respectively. About 70.2% of total NP and 24.1% of total OP were finally eliminated from water phase to surrounding matrix in the downstream. The results suggested that the downstream river channel served as the net sink of APs in the study area. PMID- 19683326 TI - Expression of the genes siamois, engrailed-2, bmp4 and myf5 during Xenopus development in presence of the marine toxins okadaic acid and palytoxin. AB - The present investigation examines the effects of the marine toxins, okadaic acid (OA) and palytoxin (PTX), on some genes involved in the neural and muscular specification and patterning of Xenopus laevis. The RT-PCR analyses performed at different stages of embryonic and larval development (stages 11-47) demonstrated that both toxins induce an over-expression of the genes siamois and engrailed-2 and a different behaviour in bmp4 and myf5. Indeed, OA provoked a significant increase in bmp4 in the earliest stage (11) examined, a down-regulation from stages 12 to 17, and a renewed increase from the beginning of hatching onwards (stages 35-47). In contrast, myf5 was up-regulated in all stages up to 35. PTX induced an over-expression of both bmp4 and myf5 during the embryonic and early larval development stages. The results show that PTX induces an increase in expression levels in all tested genes, while the response to OA seems to be more stage-dependent, with the embryonic development stage more sensitive to the toxin than the larval stages. PMID- 19683327 TI - Ecotoxicological multilevel-evaluation of the effects of fenbendazole exposure to Chironomus riparius larvae. AB - Veterinary antibiotics may find their way into the aquatic environment through direct or indirect pathways due to their widespread use. Fenbendazole is a benzimidazole anthelmintic that is widely used in veterinary medicine. To evaluate the potential ecological risk of fenbendazole, we examined the molecular and biochemical responses of biomarker genes such as heat shock proteins (HSPs), cytochrome P450 (CYP450), glutathione S-transferases (GSTs) and hemoglobins (Hbs) in Chironomus riparius for long periods. The expression of HSP70, HSP40, HSP90 and CYP450 in C. riparius increased significantly after exposure to all concentrations of fenbendazole evaluated, while the levels of GST and HbA only increased in C. riparius exposed to relatively high concentrations of fenbendazole (30 microg L(-1)). HbB expression did not differ significantly between the control and treatment groups. Exposure to 30 microg L(-1) fenbendazole had significant effects on the survival, growth, sex balance of emergent adults and development of mouthpart deformity in C. riparius. These results should constitute an important contribution to the understanding of the toxicology of fenbendazole in C. riparius. Moreover, the responses of the biomarker genes also provide valuable information that will aid in understanding the effects of fenbendazole in aquatic ecosystems. PMID- 19683328 TI - Influence of pH on the sonolysis of ciprofloxacin: Biodegradability, ecotoxicity and antibiotic activity of its degradation products. AB - The presence of antibiotics in the aquatic environment has raised concerns due to the potential risk for the emergence or persistence of antibiotic resistance. Antibiotics are often poorly degraded in conventional wastewater treatment plants. In this study, sonolysis at 520 kHz and 92 W L(-1) was used for the degradation of the fluoroquinolone antibiotic ciprofloxacin. In a first experiment at pH 7, 57% of the ciprofloxacin (15 mg L(-1)) was degraded after 120 min of ultrasonic irradiation at 25 degrees C. pH proved to be an important parameter determining the degradation rate, since the pseudo first order degradation constant increased almost fourfold when comparing treatment at pH 7 (0.0058 min(-1)) and pH 10 (0.0069 min(-1)) with that at pH 3 (0.021 min(-1)). This effect can be attributed to the degree of protonation of the ciprofloxacin molecule. The BOD/COD ratio of the solutions, which is a measure for their biodegradability, increased from 0.06 to 0.60, 0.17, and 0.18 after 120 min of irradiation depending on the pH (3, 7, and 10, respectively). The solution treated at pH 3 can even be considered readily biodegradable (BOD/COD>0.4). The antibiotic activity against Escherichia coli (G-) and Bacillus coagulans (G+) of the treated solutions also reduced after sonolysis. The highest decrease was again found when irradiated at pH 3. In contrast, ecotoxicity of the solutions to the alga Pseudokirchneriella subcapitata increased 3- to 10-fold after 20 min of treatment, suggesting the formation of toxic degradation products. The toxicity slowly diminished during further treatment. PMID- 19683329 TI - Localized insulin-derived amyloidosis in patients with diabetes mellitus: a case report. AB - Three patients with insulin dependent type 1 diabetes mellitus and one with insulin dependent type 2 diabetes mellitus developed localized amyloid tumors at their general insulin injection sites. All 4 patients (two women and two men) were treated with modern recombinant human insulin or insulin analogues. In addition, 1 patient had used both recombinant and animal insulin. The amyloid tumors were resected and examined histologically using Congo red staining and immunohistochemistry. Insulin was found to be the major component of the amyloid tumors in all four patients. These 4 cases were diagnosed recently within a relatively short period of time, which leads to the conjecture that local insulin derived amyloid tumors remain principally a differential diagnosis of skin tumors in insulin-dependent diabetic patients. PMID- 19683330 TI - Metallothionein overexpression and its prognostic relevance in intrahepatic cholangiocarcinoma and extrahepatic hilar cholangiocarcinoma (Klatskin tumors). AB - Metallothionein is a group of small molecular weight cysteine-rich proteins with a broad variety of functions. Metallothionein has been shown to regulate apoptosis and proliferation. Overexpression of metallothionein frequently occurs in human tumors and is related to prognosis as well as therapy response. However, metallothionein expression and its clinical relevance in cholangiocarcinoma have not been investigated. The present study aimed to analyze metallothionein over expression and its possible prognostic impact in intrahepatic cholangiocarcinoma and hilar extrahepatic cholangiocarcinoma (Klatskin tumors). We investigated the relationship of immunohistochemically demonstrated metallothionein expression with various clinicopathological parameters in a series of 56 intrahepatic and 56 extrahepatic cholangiocarcinoma. In noncancerous bile duct epithelia metallothionein was only occasionally weakly expressed; strong metallothionein overexpression (>50% metallothionein -positive tumor cells) was noted in 7 (12.5%) of 56 intrahepatic cholangiocarcinoma and 14 (25%) of 56 Klatskin tumors, which was associated with poor clinical outcome in univariate Kaplan-Meier testing in both intrahepatic cholangiocarcinoma (P = .002) and Klatskin tumors (P = .034). Moreover, strong metallothionein expression was identified as an independent prognostic parameter in multivariate Cox regression analysis in both intrahepatic cholangiocarcinoma (P = .005) and Klatskin tumors (P = .035). In contrast, cholangiocarcinoma with a papillary phenotype (8/112; 7.1%) exhibited a significant lack of strong metallothionein expression in all 8 of 8 cases. Strong metallothionein expression is identified as an independent poor prognostic parameter, and determination of the metallothionein expression may serve as an additional tool for the therapeutic management of patients with cholangiocarcinoma. In comparison, lack of metallothionein expression seems to be associated with cholangiocarcinoma with a papillary phenotype, which is generally recognized to have a better prognosis. PMID- 19683331 TI - A pathological reassessment of organ-confined, Gleason score 6 prostatic adenocarcinomas that progress after radical prostatectomy. AB - Prior studies of radical prostatectomies have reported a small percentage of men with biochemical progression after radical prostatectomy showing organ-confined, Gleason Score 6. One might predict that this should virtually never occur. We identified 2551 (1983-2005) radical prostatectomies coded by the urologists at our institution as pathologically organ-confined, Gleason score 6 cancer with more than 1 year of follow-up. We re-examined histopathogically the serially sectioned and completely embedded radical prostatectomy specimens of 38 men who developed biochemical recurrence defined as a single prostate-specific antigen level of 0.2 ng/mL or greater. In 27 (71%) of 38 of cases, pathology re-review showed higher grade or stage than coded by the urologists. These included 10 cases of organ-confined with Gleason pattern 4 as either the primary or secondary pattern; 9 cases of organ-confined, Gleason score 6 with tertiary pattern 4 (in 4 cases, tertiary pattern 4 was described in the initial pathology report); 5 cases of Gleason score 7 plus extraprostatic extension; 1 case of Gleason score 6 with focal extraprostatic extension; and 2 cases with positive margins due to intraprostatic incision (listed in the initial pathology report). The remaining 11 cases were true organ-confined, Gleason score 6 tumors, but none of the patients developed systemic disease. Most prior reports of organ-confined, Gleason score 6 with progression are undergraded (upgrading with revision of Gleason system), understaged (difficulty recognizing focal extraprostatic extension), or suffer from situations with ambiguous staging (intraprostatic incision) or grading (tertiary pattern 4 or 2 + 4 = 6). Even for the rare true organ-confined, Gleason score 6 (no pattern 4) tumor with supposed biochemical progression, some may be false-positive progression based on low post-radical prostatectomy prostate-specific antigen levels and minute tumors that seem highly improbable to progress. With accurate pathologic evaluation, men with organ confined, Gleason score 6 (no pattern 4) prostate cancer can be told that their risk of progression is very rare (0.4%). PMID- 19683332 TI - Anthropogenic impacts in North Poland over the last 1300 years--a record of Pb, Zn, Cu, Ni and S in an ombrotrophic peat bog. AB - Lead pollution history over Northern Poland was reconstructed for the last ca. 1300 years using the elemental and Pb isotope geochemistry of a dated Polish peat bog. The data show that Polish Pb-Zn ores and coal were the main sources of Pb, other heavy metals and S over Northern Poland up until the industrial revolution. After review of the potential mobility of each element, most of the historical interpretation was based on Pb and Pb isotopes, the other chemical elements (Zn, Cu, Ni, S) being considered secondary indicators of pollution. During the last century, leaded gasoline also contributed to anthropogenic Pb pollution over Poland. Coal and Pb-Zn ores, however, remained important sources of pollution in Eastern European countries during the last 50 years, as demonstrated by a high (206)Pb/(207)Pb ratio (1.153) relative to that of Western Europe (ca. 1.10). The Pb data for the last century were also in good agreement with modelled Pb inventories over Poland and the Baltic region. PMID- 19683333 TI - Time allocation shifts and pollutant exposure due to traffic congestion: an analysis using the national human activity pattern survey. AB - Traffic congestion increases air pollutant exposures of commuters and urban populations due to the increased time spent in traffic and the increased vehicular emissions that occur in congestion, especially "stop-and-go" traffic. Increased time in traffic also decreases time in other microenvironments, a trade off that has not been considered in previous time activity pattern (TAP) analyses conducted for exposure assessment purposes. This research investigates changes in time allocations and exposures that result from traffic congestion. Time shifts were derived using data from the National Human Activity Pattern Survey (NHAPS), which was aggregated to nine microenvironments (six indoor locations, two outdoor locations and one transport location). After imputing missing values, handling outliers, and conducting other quality checks, these data were stratified by respondent age, employment status and period (weekday/weekend). Trade-offs or time-shift coefficients between time spent in vehicles and the eight other microenvironments were then estimated using robust regression. For children and retirees, congestion primarily reduced the time spent at home; for older children and working adults, congestion shifted the time spent at home as well as time in schools, public buildings, and other indoor environments. Changes in benzene and PM(2.5) exposure were estimated for the current average travel delay in the U.S. (9 min day(-1)) and other scenarios using the estimated time shifts coefficients, concentrations in key microenvironments derived from the literature, and a probabilistic analysis. Changes in exposures depended on the duration of the congestion and the pollutant. For example, a 30 min day(-1) travel delay was determined to account for 21+/-12% of current exposure to benzene and 14+/-8% of PM(2.5) exposure. The time allocation shifts and the dynamic approach to TAPs improve estimates of exposure impacts from congestion and other recurring events. PMID- 19683335 TI - Phosphorylation of alpha-synuclein upregulates tyrosine hydroxylase activity in MN9D cells. AB - Hyperphosphorylated alpha-synuclein is considered an important event in the pathogenesis of Parkinson's disease but its function remains elusive. In this study we provide evidence that tyrosine hydroxylase (TH) expression was unaffected by overexpression of wild-type and phospho-mimic mutant alpha synuclein (S129D) in dopaminergic MN9D cells. However, alpha-synuclein overexpression evidently inhibited TH phosphorylation at Ser40 and dopamine synthesis, while alpha-synuclein (S129D) mutant enhanced TH phosphorylation and dopamine synthesis. This phospho-mimic mutant prevented wild-type alpha-synuclein cytotoxicity to MN9D cells, which might be due to aggregation of mutant alpha synuclein in the cytoplasm and nuclei. These results demonstrated that phosphorylation at Ser129 was involved in the regulation of TH activity, as well as in eliminating the neurotoxicity of wild-type alpha-synuclein overexpression in MN9D cells. PMID- 19683334 TI - Innate immunity, coagulation and placenta-related adverse pregnancy outcomes. AB - Maternal immunity undergoes subtle adjustment in order to tolerate the semi allogeneic embryo and maintain the host defense against potential pathogens. Concomitantly, coagulation systems change from an anti-coagulant state to a pro coagulant state to meet the hemostatic challenge of placentation and delivery. Innate immunity and blood coagulation systems are the first line of defense to protect a host against exogenous challenges, including alloantigens and mechanical insults, and preserve the integrity of an organism. The interactions between coagulation and immune systems have been extensively studied. Immune cells play a pivotal role in the initiation of the coagulation cascade, whereas coagulation proteases display substantial immuno-modulatory effects. Upon exogenous challenges, the immune and coagulation systems are capable of potentiating each other leading to a vicious cycle. Natural killer (NK) cells, macrophages (Mphis) and dendritic cells (DCs) are three major innate immune cells that have been demonstrated to play essential roles in early pregnancy. However, immune maladaptation and hemostatic imbalance have been suggested to be responsible for adverse pregnant outcomes, such as preeclampsia (PE), miscarriage, recurrent spontaneous abortion (RSA) and intrauterine growth restriction (IUGR). In this review, we will summarize the mutual regulation between blood coagulation and innate immune systems as well as their roles in the maintenance of normal pregnancy and in the pathogenesis of adverse pregnancy outcomes. PMID- 19683337 TI - Duration of spermatogenesis in the bullfrog (Lithobates catesbeianus). AB - The bullfrog (Lithobates catesbeianus) has substantial economic importance and has also been used as an experimental model for biological studies in the fields of pharmacology, medicine, and reproductive biology, especially studies addressing gametogenesis. However, there is a lack of comprehensive information in the literature regarding testis structure and function in this amphibian. The main objective of the current study was to estimate the duration of the various phases of spermatogenesis in this vertebrate. Sixteen sexually mature bullfrogs received an intracoelomic administration of tritiated thymidine. Testes were analyzed at various times between 1h and 33 d after administration to detect the most advanced germ cell types labeled at each interval, as well as labeled preleptotene spermatocytes, which presumably originated from spermatogonial stem cells. The duration of the spermatogonial, spermatocytic, and spermiogenic phases of spermatogenesis in the bullfrog were approximately 18, 14, and 8 d, respectively. Thus, the total duration of the spermatogenesis process from early spermatogonia through to spermatozoa was 40 d in this species, similar to that of most previously investigated mammalian species. To our knowledge, this is the first reliable report on the duration of the full spermatogenic process in any amphibian species. These findings will be very useful for tracking the pace of germ cells in studies involving spermatogonial transplantation in lower vertebrates. PMID- 19683338 TI - Mechanical behavior of facial prosthetic elastomers after outdoor weathering. AB - OBJECTIVES: The degradation of maxillofacial prosthetic elastomers that occurs during physical weathering is usually responsible for the replacement of prostheses. In this study the mechanical behavior of 4 non-pigmented facial prosthetic elastomers, exposed to outdoor weathering for 1 year, was investigated. The hypothesis investigated was that irradiation time did not affect the properties measured. METHODS: The samples were exposed to solar radiation for 1 year in Thessaloniki (Greece). Three different types of polydimethyl siloxane (PDMS) and chlorinated polyethylene (CPE) samples were tested in this study. Mechanical tests (compressive-tensile) were performed using a universal type testing machine. Hardness tests were evaluated using a durometer tester. Simple mathematical models were developed to correlate the measured properties with irradiation time. The stress-strain data of compression and tensile tests were modeled using parameters such as maximum stress (sigma(max)), maximum strain (epsilon(max)), elasticity parameter (E), and non-linearity parameter (p), while the mathematical model used for hardness data involves initial hardness of materials (H(0)). RESULTS: Two of the silicone prosthetics (Elastomer 42, TechSil 25) seem to become harder and more brittle contrary to the other silicone (M 511) and chlorinated polyethylene (CPE) samples that become softer and more ductile. Duration of exposure increases these phenomena. CONCLUSION: The effect of irradiation time on the mechanical behavior was introduced through its effect on the models' parameters. The hypothesis was rejected since changes were observed in the model parameters. PMID- 19683339 TI - Profiles of British Muslim identity: adolescent girls in Birmingham. AB - By asking students to fill in 10 statements beginning with 'I am...' and a further 10 statements beginning with 'I am not...' we constructed profiles of British Muslim ethnic and national identity. Participants were 108 British Muslim girls of mean age 12.6 years studying in a single sex girls' school in Birmingham, UK. Using content analysis we found that some adolescent girls are rooted only in their personal identity. Others are rooted in their social identity. We found a number of major categories: rooted in ethnic identity only, rooted in ethnic identity but differentiating self from other groups, rooted in bicultural identity, rooted in national identity and confused ethnic and national identity. Significantly a number of participants were concerned to transcend group identities and described themselves as 'not racist' or 'not prejudiced'. A model of ethnic and national identity is presented incorporating rootedness, differentiation, confusion and transcendence. PMID- 19683336 TI - Comel-Netherton syndrome defined as primary immunodeficiency. AB - BACKGROUND: Mutations in serine protease inhibitor Kazal-type 5 (SPINK5), encoding the serine protease inhibitor lympho-epithelial Kazal-type 5 related inhibitor (LEKTI), cause Comel-Netherton syndrome, an autosomal-recessive disease characterized by congenital ichthyosis, bamboo hair, and atopic diathesis. Despite increased frequency of infections, the immunocompetence of patients with Comel-Netherton syndrome has not been extensively investigated. OBJECTIVE: To define Comel-Netherton syndrome as a primary immunodeficiency disorder and to explore the benefit of intravenous immunoglobulin replacement therapy. METHODS: We enrolled 9 patients with Comel-Netherton syndrome, sequenced SPINK5, and analyzed LEKTI expression by immunohistochemistry. Immune function was assessed by measuring cognate immunity, serum cytokine levels, and natural killer cell cytotoxicity. RESULTS: All patients presented with recurrent skin infections caused predominantly by Staphylococcus aureus. All but 1 reported recurrent respiratory tract infections; 78% had sepsis and/or pneumonia; 67% had recurrent gastrointestinal disease and failure to thrive. Mutations in SPINK5-including 6 novel mutations-were identified in 8 patients. LEKTI expression was decreased or absent in all patients. Immunologic evaluation revealed reduced memory B cells and defective responses to vaccination with Pneumovax and bacteriophage phiX174, characterized by impaired antibody amplification and class-switching. Immune dysregulation was suggested by a skewed T(h)1 phenotype and elevated proinflammatory cytokine levels, whereas serum concentrations of the chemokine (C C motif) ligand 5 and natural killer cell cytotoxicity were decreased. Treatment with intravenous immunoglobulin resulted in remarkable clinical improvement and temporarily increased natural killer cell cytotoxicity. CONCLUSION: These data provide new insights into the immunopathology of Comel-Netherton syndrome and demonstrate that this multisystem disorder, characterized by lack of LEKTI expression in epithelial cells, is complicated by cognate and innate immunodeficiency that responds favorably to intravenous immunoglobulin therapy. PMID- 19683340 TI - Peptic ulcer disease. AB - Peptic ulcer disease had a tremendous effect on morbidity and mortality until the last decades of the 20th century, when epidemiological trends started to point to an impressive fall in its incidence. Two important developments are associated with the decrease in rates of peptic ulcer disease: the discovery of effective and potent acid suppressants, and of Helicobacter pylori. With the discovery of H pylori infection, the causes, pathogenesis, and treatment of peptic ulcer disease have been rewritten. We focus on this revolution of understanding and management of peptic ulcer disease over the past 25 years. Despite substantial advances, this disease remains an important clinical problem, largely because of the increasingly widespread use of non-steroidal anti-inflammatory drugs (NSAIDs) and low-dose aspirin. We discuss the role of these agents in the causes of ulcer disease and therapeutic and preventive strategies for drug-induced ulcers. The rare but increasingly problematic H pylori-negative NSAID-negative ulcer is also examined. PMID- 19683341 TI - Cholesterol-mediated anchoring of enzyme-loaded liposomes within disulfide stabilized polymer carrier capsules. AB - Polymer capsules containing multiple liposomes, termed capsosomes, are a promising new concept toward the design of artificial cells. Herein, we report on the fundamental aspects underpinning the assembly of capsosomes. A stable and high loading of intact liposomal cargo into a polymer film was achieved by non covalently sandwiching the liposomes between a tailor-made cholesterol-modified poly(L-lysine) (PLL(c)) precursor layer and a poly(methacrylic acid)-co (cholesteryl methacrylate) (PMA(c)) capping layer. The film assembly, optimized on planar surfaces, was successfully transferred onto colloidal substrates, and a polymer membrane was subsequently assembled by the alternating adsorption of poly(N-vinyl pyrrolidone) (PVP) and thiol-modified poly(methacrylic acid) (PMA(SH)) onto the pre-adsorbed layer of liposomes. Upon removal of the silica template, stable capsosomes encapsulating the enzyme luciferase or beta-lactamase within their liposomal sub-compartments were obtained at both assembly (pH 4) and physiological conditions (pH 7.4). Excellent retention of the liposomes and the enzymatic cargo within the polymer carrier capsules was observed for up to 14 days. These engineered capsosomes are particularly attractive as autonomous microreactors, which can be utilized to repetitively add smaller reactants to cause successive distinct reactions within the capsosomes and simultaneously release the products to the surrounding environment, bringing these systems one step closer toward constructing artificial cells. PMID- 19683343 TI - Bioaccumulation and biotransformation of brominated and chlorinated contaminants and their metabolites in ringed seals (Pusa hispida) and polar bears (Ursus maritimus) from East Greenland. AB - We report on the comparative bioaccumulation, biotransformation and/or biomagnification from East Greenland ringed seal (Pusa hispida) blubber to polar bear (Ursus maritimus) tissues (adipose, liver and brain) of various classes and congeners of persistent chlorinated and brominated contaminants and metabolic by products: polychlorinated biphenyls (PCBs), chlordanes (CHLs), hydroxyl (OH-) and methylsulfonyl (MeSO(2)-) PCBs, polybrominated biphenyls (PBBs), OH-PBBs, polybrominated diphenyl ether (PBDE) and hexabromocyclododecane (HBCD) flame retardants and OH- and methoxyl (MeO-) PBDEs, 2,2-dichloro-bis(4 chlorophenyl)ethene (p,p'-DDE), 3-MeSO(2)-p,p'-DDE, pentachlorophenol (PCP) and 4 OH-heptachlorostyrene (4-OH-HpCS). We detected all of the investigated contaminants in ringed seal blubber with high frequency, the main diet of East Greenland bears, with the exception of OH-PCBs and 4-OH-HpCS, which indicated that these phenolic contaminants were likely of metabolic origin and formed in the bears from accumulated PCBs and octachlorostyrene (OCS), respectively, rather than being bioaccumulated from a seal blubber diet. For all of the detectable sum of classes or individual organohalogens, in general, the ringed seal to polar bear mean BMFs for SigmaPCBs, p,p'-DDE, SigmaCHLs, SigmaMeSO(2)-PCBs, 3-MeSO(2) p,p'-DDE, PCP, SigmaPBDEs, total-(alpha)-HBCD, SigmaOH-PBDEs, SigmaMeO-PBDEs and SigmaOH-PBBs indicated that these organohalogens bioaccumulate, and in some cases there was tissue-specific biomagnification, e.g., BMFs for bear adipose and liver ranged from 2 to 570. The blood-brain barrier appeared to be effective in minimizing brain accumulation as BMFs were 200 (CD4 200-499: OR 0.5, 95% CI 0.3-0.9. CD4> or =500: OR 0.3, 95% CI 0.2-0.6), and contact with persons with TB (OR 0.2, 95% CI 0.1-0.5) were associated with a lower probability. CONCLUSIONS: In this study, the percentage of TST non prescription was quite high. The results suggest that TST non-prescription in this population is related to the clinicians' expectations regarding the results of the test and the patients' adherence to treatment for latent TB infection. PMID- 19683365 TI - [Asymptomatic osteonecrosis of the hip in HIV-infected patients]. AB - BACKGROUND: Osteonecrosis (ON) of the hip in human immunodeficiency virus (HIV) infected patients has been related to corticosteroid use, dyslipidemia, alcoholism, lipodystrophy, antiretroviral drug use, and HIV infection, itself. The aim of this study was to evaluate the prevalence of silent ON of the hip in HIV-infected patients and its association with several risk factors. METHODS: From a total of 1200 patients followed up at the HIV clinic of Xeral-Cies Hospital in Vigo (Spain), we selected those diagnosed with HIV infection before January 2006, aged 20-70 years, who came to the clinic during the period of March to May 2008. Patients with a diagnosis of ON of the hip and those with current symptoms were excluded. A magnetic resonance imaging (MRI) study of the hips was performed. RESULTS: Ninety-seven Caucasian patients (68 men) with a median age of 44 years were included. ON of the hip was detected in 4 patients (4.1%), with bilateral involvement in 2 patients. All 4 patients were men, with a median age of 44.5 years. Several risk factors for ON of the hip were recorded in each patient. CD4 lymphocyte nadir (P=0.034), percentage of patients with CDC stage C (P=0.039), and number of patients with previous corticosteroid treatment (P=0.042) were significantly different between patients with ON of the hip and those with normal MRI findings. CONCLUSIONS: The prevalence of asymptomatic ON of the hip in HIV-infected patients was 4.1%. The most important risk factors for developing this condition were corticosteroid treatment, lower CD4 lymphocyte nadir, and AIDS-defining disease. Antiretroviral treatment was not associated with osteonecrosis. PMID- 19683366 TI - Cytotoxicity and effects of 1,10-phenanthroline and 5-amino-1,10-phenanthroline derivatives on some immunocompetent cells. AB - The biological activity of previously synthesized compounds [(phen)(3)(H(+))(2)(NO(3)(-))(2) (1), Pd(5-NH(2)-phen)(2)(NO(3))(2) (2) and Pd(phen)(2)(NO(3))(2)(H(2)O) (3)] was investigated in vivo. The three compounds did not show any histological alterations in the observed lung, liver, spleen and lymph nodes of White Wistar rats. The propidium iodine staining did not discover any cytotoxic effect of the tested derivatives. The tests for immunological response predominantly showed stimulation of the antibody-producing B-cells and lower or no stimulation of the T-cells. The LIF-test showed better stimulation of all lymphocytes with 1, followed by 2 and 3. Substance 3 showed highest stimulating effect on B-cell blood lymphocytes in all doses (maximum in the lowest dose), whereas the impact of 2 is weaker and that of 1 is the weakest. The T-cell immune response after treatment with substance 1 is best influenced by dose of 1mg in the spleen cell-fraction, followed by 3 (5 mg). PMID- 19683367 TI - [The Revue de Medecine Interne is moving on!]. PMID- 19683368 TI - [What's new in the treatment of obesity?]. PMID- 19683369 TI - [Polymyositis and dermatomyositis in adults. Tunisian multicentre study]. AB - INTRODUCTION: To study the clinical features and the outcome of polymyositis (PM) and dermatomyositis (DM) in Tunisia. METHODS: Retrospective multicentre study conducted by the Tunisian Society of Internal Medicine. Adult patients with confirmed and probable PM and DM, according to the Bohan and Peter's criteria, were included in the study and followed-up between 1987 and 2005. RESULTS: Twenty cases of PM and 50 cases of DM were included. Female/male sex-ratio was 2.5. The mean age at diagnosis was 40.7 years. Muscle weakness concerned girdle muscles in 94.3% and pharyngeal muscles in 42.8% of the patients. Interstitial lung disease and heart involvement occurred respectively in 35.7 and 20%. Eyelid erythema, peri-ungual erythema and Gottron's sign occurred in respectively 86, 50 and 36% of the cases of DM. Malignant disease was found in 12.8% of the patients (mainly gynecological and nasopharyngeal cancers). All patients received high dose corticosteroids with additional immunosuppressive drugs in 40% of the cases. The outcome was stabilization or partial improvement in 71.5% of the cases. Death occurred in 20% and was caused by respiratory failure, sepsis and cardiac failure. CONCLUSION: The particularities of PM and DM in Tunisia are the preponderance of DM, early onset of the disease and high frequency of the nasopharyngeal cancer. PMID- 19683370 TI - Validation of skinfold measurements and bioelectrical impedance analysis in children with severe cerebral palsy: a review. AB - BACKGROUND & AIMS: Accurately measuring nutritional status in children with severe cerebral palsy (CP) is a challenge. This review seeks to assess the validity of skinfold measurements and bioelectrical impedance analysis (BIA) for measuring body composition in children with severe CP. METHODS: We executed a literature search on the validation of both methods in children with severe CP. To be eligible for inclusion, a study had to report on a statistical comparison between these two methods and any method of reference. The QUADAS tool was used for quality assessment. RESULTS: The search strategy resulted in 1549 studies of which 5 studies eventually met the inclusion criteria. When comparing body composition outcomes of skinfold measurements to a reference method, correlation coefficients were found ranging from 0.406 to 0.988. Correlation coefficients between body composition data of BIA and a reference method ranged from 0.515 to 0.95. CONCLUSIONS: Although a number of authors found favorable agreement between skinfold measurements and BIA in comparison with reference methods, the small numbers studied, the lack of methodological quality measured by QUADAS, and the use of inappropriate analytical methods hamper solid conclusions. PMID- 19683371 TI - Merkel cell carcinoma of the face: follow-up report of 10 cases in Germany. AB - Merkel cell carcinoma is a rare cutaneous neoplasm in older adults that has a high propensity for nodal metastases and local recurrence, has poor prognosis, and in the last few years has increased in incidence. We retrospectively review our experience of the surgical treatment of Merkel cell carcinoma of the face in 10 patients (9 women and 1 man) between 1993 and 2007. Follow-up ranged from 5 to 132 months (mean 38.8, median 29). Relapse-free survival for all patients was 47% at 2 years. We support the need for complete excision with wide 2-3cm margins throughout all tumour stages although this might be limited when it affects the face. We also recommend radiotherapy for patients with disease that is stage II and above; the value of adjuvant radiotherapy is outlined in many studies. Therapeutic functional neck dissection should be done in cases of nodal disease. The role of chemotherapy is still not clear, and it has not been used in our department. PMID- 19683373 TI - Knowledge explosion in phytotechnologies for environmental solutions. AB - Bioremediation using biotechnology and biodiversity--a multiscale approach. PMID- 19683372 TI - Investigation of standing balance in diabetic patients with and without peripheral neuropathy using accelerometers. AB - BACKGROUND: Peripheral neuropathy is recognised to be the most symptomatic complication of diabetes and is also linked to postural instability. This study investigates balance instability in diabetic patients with and without peripheral neuropathy. METHODS: Quiet standing balance was investigated using an accelerometric-based method in 24 diabetic patients (12 with and 12 without peripheral neuropathy) and compared with 12 control subjects. Accelerations were measured at lumbar and ankle levels using three accelerometers. Two standing conditions of 30s were evaluated (i.e., eyes opened, eyes closed). The range and root mean square values were calculated on the anterior posterior component of lumbar and ankle accelerations and for the medial lateral component of lumbar accelerations. Differences between parameters were compared between groups using ANOVA and post hoc comparisons. FINDINGS: The diabetic patients with peripheral neuropathy show higher-range and root mean square values compared with those of control subjects and diabetic patients without peripheral neuropathy. Significant differences between groups have been detected for anterior posterior range of lumbar acceleration, which was significantly higher for diabetic patients with peripheral neuropathy, compared with those of others groups. Significant higher values for diabetic patients with peripheral neuropathy were also detected for anterior posterior range and root mean square of ankle accelerations compared with control subjects. Visual deprivation shows an increase in accelerometric parameters for each group. INTERPRETATION: This study is the first to investigate the balance instability of diabetic patients using accelerometers. Results confirm that diabetic patients with peripheral neuropathy have greater postural instability with higher acceleration values than those of control group and diabetic patients without peripheral neuropathy. PMID- 19683374 TI - Mobility of arsenic, cadmium and zinc in a multi-element contaminated soil profile assessed by in-situ soil pore water sampling, column leaching and sequential extraction. AB - Three methods for predicting element mobility in soils have been applied to an iron-rich soil, contaminated with arsenic, cadmium and zinc. Soils were collected from 0 to 30 cm, 30 to 70 cm and 70 to 100 cm depths in the field and soil pore water was collected at different depths from an adjacent 100 cm deep trench. Sequential extraction and a column leaching test in the laboratory were compared to element concentrations in pore water sampled directly from the field. Arsenic showed low extractability, low leachability and occurred at low concentrations in pore water samples. Cadmium and zinc were more labile and present in higher concentrations in pore water, increasing with soil depth. Pore water sampling gave the best indication of short term element mobility when field conditions were taken into account, but further extraction and leaching procedures produced a fuller picture of element dynamics, revealing highly labile Cd deep in the soil profile. PMID- 19683375 TI - Quantification of activated carbon contents in soils and sediments using chemothermal and wet oxidation methods. AB - Activated carbon (AC) strongly sorbs organic pollutants and can be used for remediation of soils and sediments. A method for AC quantification is essential to monitor AC (re)distribution. Since AC is black carbon (BC), two methods for BC quantification were tested for AC mixed in different soils and sediments: i) chemothermal oxidation (CTO) at a range of temperatures and ii) wet-chemical oxidation with a potassium dichromate/sulfuric acid solution. For three soils, the amount of AC was accurately determined by CTO at 375 degrees C. For two sediments, however, much of the AC disappeared during combustion at 375 degrees C, which could probably be explained by catalytic effects by sediment constituents. CTO at lower temperatures (325-350 degrees C) was a feasible alternative for one of the sediments. Wet oxidation effectively functioned for AC quantification in sediments, with almost complete AC recovery (81-92%) and low remaining amounts of native organic carbon (5-16%). PMID- 19683376 TI - Moral injury and moral repair in war veterans: a preliminary model and intervention strategy. AB - Throughout history, warriors have been confronted with moral and ethical challenges and modern unconventional and guerilla wars amplify these challenges. Potentially morally injurious events, such as perpetrating, failing to prevent, or bearing witness to acts that transgress deeply held moral beliefs and expectations may be deleterious in the long-term, emotionally, psychologically, behaviorally, spiritually, and socially (what we label as moral injury). Although there has been some research on the consequences of unnecessary acts of violence in war zones, the lasting impact of morally injurious experience in war remains chiefly unaddressed. To stimulate a critical examination of moral injury, we review the available literature, define terms, and offer a working conceptual framework and a set of intervention strategies designed to repair moral injury. PMID- 19683377 TI - Bonding effectiveness of two contemporary self-etch adhesives to enamel and dentin. AB - OBJECTIVES: Among contemporary adhesives, self-etch adhesives have been adopted by general practitioners for routine adhesive restorative purposes, mainly because of their ease of use. However, many versions that differ for their clinical application procedure, pH, number of components, etc., are currently available on the market. The purpose of this study was to determine the bonding effectiveness of two new self-etch adhesives (Adper Easy Bond and Adper ScotchBond SE, 3M ESPE) to enamel and dentin using a micro-tensile bond strength (microTBS) protocol and to characterise the interfacial ultra-structure at enamel and dentin using transmission electron microscopy (TEM). METHODS: The adhesives were applied onto coronal human enamel and dentin surfaces and built up with the micro-hybrid resin composite Z100 (3M ESPE). The 'gold-standard' two-step self etch adhesive Clearfil SE Bond (Kuraray) served as control. Specimens were sectioned to sticks and trimmed at the interface to a cylindrical hour-glass shape ('trimmed' micro-specimens). Non-demineralized and demineralized TEM sections through the adhesive-dentin/enamel interface were prepared by ultra microtomy. RESULTS: The microTBS of the two self-etch adhesives to enamel was statistically significantly lower than that of the control. To dentin, the microTBS of Adper Easy Bond was significantly lower than that of Adper ScotchBond SE and the control. TEM showed a tight interface to enamel for all three self etch adhesives. A relatively thick, completely demineralized and acid-resistant hybrid layer was formed at dentin by Adper ScotchBond SE, whereas the interaction of Adper Easy Bond was much shallower, and comparable to that of so-called 'ultra mild' self-etch adhesives. Some degree of spot- and cluster-like nano-leakage was observed for both adhesives, but did not differ in extent and form from that observed for the control. CONCLUSIONS: Although the new two self-etch adhesives revealed a tight interaction at both enamel and dentin, their bond strength to both tooth tissues was generally lower than that of the control adhesive. Nevertheless, their bonding effectiveness appears in line with other simplified self-etch adhesives. PMID- 19683378 TI - A preliminary evaluation of the structural integrity and fracture mode of minimally prepared resin bonded CAD/CAM crowns. AB - PURPOSE: This study was a preliminary evaluation of two minimal preparation designs proposed for ceramic and composite resin bonded CAD/CAM crowns. It compared the structural integrity and fracture mode of teeth restored with traditionally and minimally prepared resin bonded CAD/CAM crowns fabricated from the same material hypothesizing that teeth restored with minimal resin bonded crowns would demonstrate the same fracture strength to teeth restored with traditional resin bonded crowns. MATERIALS AND METHODS: Forty intact maxillary molar teeth were used and divided in four groups. Two groups were prepared according to a traditional crown preparation design and two groups were prepared according to minimal preparation designs. A resin composite (Paradigm MZ100, 3M ESPE) and a leucite glass-ceramic (ProCAD, Ivoclar Vivadent) were used for the fabrication of the crowns using CEREC Scan. Crowns of ceramic were cemented using Variolink II (Ivoclar Vivadent) and crowns of composite with Rely X Unicem Aplicap (3M ESPE) and loaded until fracture. Load data was analysed using ANOVA comparing crowns of the same restorative material. The mode of fracture was also recorded and analysed (Kruskal-Wallis). RESULTS: For the composite system the mean fracture load and SD was 1682N (+/-315) for the traditional and 1751N (+/ 338) for the minimal crowns. For the ceramic system the mean fracture load and SD was 1512N (+/-373) for the traditional and 1837 (+/-356) for the minimal crowns. No statistically significant difference was found between the two designs for each system. Nonparametric analysis (Kruskal-Wallis) of the fracture mode showed no statistical significant difference between designs for either material (p>.05). CONCLUSION: Within the limitations of this experimental design, it was found that minimally prepared resin bonded CEREC crowns demonstrated equal fracture resistance and mode of fracture to that of crowns bonded to traditionally prepared teeth. PMID- 19683379 TI - Resuscitation Great. Rudolf Juchems--a pioneer of cardiopulmonary resuscitation in Germany. PMID- 19683382 TI - Open to debate. The motion: PDE5 inhibitors are a promising therapy for benign prostatic hyperplasia. PMID- 19683381 TI - In vivo quantification of liver stiffness in a rat model of hepatic fibrosis with acoustic radiation force. AB - Liver fibrosis is currently staged using needle biopsy, a highly invasive procedure with a number of disadvantages. Measurement of liver stiffness changes that accompany progression of the disease may provide a quantitative and noninvasive method to assess the health of the liver. The purpose of this study is to investigate the correlation between liver stiffness measured by radiation force induced shear waves and disease related changes in the liver. An additional aim is to present initial findings on the effects of liver viscosity on radiation force induced shear wave morphology. Liver fibrosis was induced in 10 rats using carbon tetrachloride (CCl(4)), while five rats acted as controls. Liver stiffness was measured in vivo in all rats after a treatment period of 8 weeks using a modified Siemens SONOLINE Antares scanner (Siemens Medical Solutions USA, Ultrasound Division, Issaquah, WA, USA). The spatial coherence of radiation force induced shear waves propagating in the viscoelastic rat liver decreased significantly with propagation distance, compared with shear waves in an elastic phantom and a finite element model of a purely elastic medium. Animals were sacrificed after imaging and liver samples were taken for histopathologic analysis and collagen quantification using picrosirius red staining and hydroxyproline assay. At the end of the treatment period, five rats had healthy livers (stage F0), while six had severe fibrosis (F3) and the rest had light to moderate fibrosis (F1 and F2). The measured liver stiffness for the F0 group was 1.5+/-0.1 kPa (mean+/-95% confidence interval) and for F3 livers was 1.8+/-0.2 kPa. In this study, liver stiffness was found to be linearly correlated with the amount of collagen in the liver measured by picrosirius red staining (r(2)=0.43, p=0.008). In addition, stiffness spatial heterogeneity was also linearly correlated with liver collagen content (r(2)=0.58, p=0.001) by picrosirius red staining. These results are consistent with those obtained by Salameh et al. (2007) and Yin et al. (2007b) using animal models of liver fibrosis and MR elastography. This suggests that stiffness measurement using acoustic radiation force can provide a quantitative assessment of the extent of fibrosis in the liver and can be potentially used for the diagnosis, management and study of liver fibrosis. PMID- 19683380 TI - Ultrasound phase contrast thermal imaging with reflex transmission imaging methods in tissue phantoms. AB - Thermal imaging measurements using ultrasound phase contrast have been performed in tissue phantoms heated with a focused ultrasound source. Back projection and reflex transmission imaging principles were used to detect sound speed-induced changes in the phase caused by an increase in the temperature. The temperature was determined from an empirical relationship for the temperature dependence on sound speed. The phase contrast was determined from changes in the sound field measured with a hydrophone scan conducted before and during applied heating. The lengthy scanning routine used to mimic a large two-dimensional array required a steady-state temperature distribution within the phantom. The temperature distribution in the phantom was validated with magnetic resonance (MR) thermal imaging measurements. The peak temperature was found to agree within 1 degrees C with MR, and good agreement was found between the temperature profiles. The spatial resolution was 0.3x0.3x0.3mm, comparing favorably with the 0.625x0.625x1.5-mm MR spatial resolution. PMID- 19683384 TI - Excellent response to thrombolysis following prolonged basilar artery occlusion with extensive ischemic changes on MRI. PMID- 19683385 TI - Worldwide genetic diversity of HBV genotypes and risk of hepatocellular carcinoma. AB - Hepatitis B viruses (HBV) are responsible for over 50% of the worldwide attributable risk of hepatocellular carcinoma (HCC) and this figure increases even further in regions of high endemicity. Systematic sequencing of HBV genomes has identified that this common virus existed as eight distinct genotypes (denoted A-H), each regrouping variants with less than 8% divergence in their DNA sequence. These genotypes differ by their geographic distribution in populations around the globe. There is evidence that HBV genotypes also differ by their pathogenic properties, including their risk of persistence as chronic infection and their capacity to induce precursor disease or cancer. On the other hand, HBV genes may undergo mutations that become selected during the course of chronic infection and progressive liver disease. The most significant of these mutations in the context of HCC are those occurring in the pre-core (Pre-C) and basal core promoter (BCP) regions. These mutations may upregulate HBV expression and increase its virulence. These mutations may occur in all HBV genotypes but are more common in genotypes associated with more severe disease and cancer, in particular genotype C. Understanding the molecular basis of pathological variations between HBV variants is critical for prediction of disease severity. It will also be important to determine whether differences among genotypes may have an impact on the long-term protective efficacy of universal HBV vaccination. PMID- 19683386 TI - Exploring the influence of operational parameters on the reactivity of elemental iron materials. AB - In an attempt to characterize material intrinsic reactivity, iron dissolution from elemental iron materials (Fe(0)) was investigated under various experimental conditions in batch tests. Dissolution experiments were performed in a dilute solution of ethylenediaminetetraacetate (Na(2)-EDTA - 2mM). The dissolution kinetics of 18 Fe(0) materials were investigated. The effects of individual operational parameters were assessed using selected materials. The effects of available reactive sites [Fe(0) particle size (G genetic variant for pain treatment. AB - Regard of functional pharmacogenetic polymorphisms may further the success of pain therapy by adopting individualized approaches. The mu-opioid receptor gene (OPRM1) 118A>G polymorphism is a promising candidate for both opioid effects and pain because of both biological reasonability and apparent experimental and clinical evidence. We analyzed its importance for pain therapy using a meta analytic approach to studies relating it to opioid pain therapy. Data from suitable studies selected from hits of a PubMed search for "OPRM1" were independently extracted by two authors. The meta-analysis included phenotypes by OPRM1 genotype (opioid dosing, pain, and side effects), publication year, diagnostic status, proportion of male study participants, and whether genotype frequencies agreed with Hardy-Weinberg equilibrium. We found no consistent association between OPRM1 118A>G genotypes and most of the phenotypes in a heterogeneous set of eight clinical studies. Only weak evidence of an association with less nausea (effect size, Cohen's d=-0.21, p=0.037) and of increased opioid dosage requirements (d=0.56, p=0.018) in homozygous carriers of the G allele was obtained. This indicates that despite initially promising results, available evidence of the clinical relevance of the OPRM1 118A>G polymorphism does not withhold a meta-analysis. This discourages basing personalized therapeutic concepts of pain therapy on OPRM1 118A>G genotyping at the present state of evidence. PMID- 19683392 TI - Intrathecal glycine for pain and dystonia in complex regional pain syndrome. AB - Since glycinergic neurotransmission plays an important inhibitory role in the processing of sensory and motor information, intrathecal glycine (ITG) administration may be a potential therapy for both pain and movement disorders in patients with complex regional pain syndrome (CRPS). Aims of the current study, which is the first report on ITG in humans, were to evaluate its safety and efficacy. ITG treatment during 4 weeks was studied in CRPS patients with dystonia in the period before they received intrathecal baclofen treatment. Twenty patients were assessed and after exclusion of one patient, the remaining 19 patients were randomized in a double-blind placebo-controlled crossover study. Safety was assessed by clinical evaluation, blood examinations and electrocardiograms. Efficacy measures involved pain (numeric rating scale, McGill pain questionnaire), movement disorders (Burke-Fahn-Marsden dystonia rating scale, unified myoclonus rating scale, tremor research group rating scale), activity (Radboud skills questionnaire, walking ability questionnaire), and a clinical global impression (CGI) and patient's global impression score (PGI). Treatment-emergent adverse events were generally mild to moderate and not different from placebo treatment. During ITG treatment growth hormone levels were slightly increased. Although there was a trend to worsening on the CGI and PGI during ITG treatment, there were no significant differences between ITG and placebo treatment in any of the outcomes. ITG given over 4 weeks was ineffective for pain or dystonia in CRPS. Although no serious adverse events occurred, further studies are required to rule out potential neurotoxicity of ITG. PMID- 19683394 TI - Expressive dimensions of pain catastrophizing: An observational study in adolescents with chronic pain. AB - Investigated was the relationship between pain catastrophizing and pain intensity in adolescents suffering from chronic pain (n = 38) and the extent to which they expressed communicative pain and pain-related protective behaviours. Adolescents were observed on video performing a 2-Min Walk Test (2MWT). Behaviours were coded on videotape. The adolescents' verbalizations about the 2MWT were also rated by their parents. Analyses revealed that higher levels of catastrophic thinking about pain were associated with higher levels of facial pain expressions and verbalizations about their pain experience, beyond the effects of age, gender, pain duration and pain intensity. Pain-related protective behaviours did not vary with the adolescents' level of pain catastrophizing, but varied with pain intensity. The findings corroborate the functional distinctiveness of different types of pain behaviours. The results are discussed in terms of the processes linking (1) catastrophizing to communicative pain behaviours and (2) pain to pain related protective behaviours. PMID- 19683396 TI - Comparison of the susceptibility of brown trout (Salmo trutta) and four rainbow trout (Oncorhynchus mykiss) strains to the myxozoan Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD). AB - Differences in susceptibility to the myxozoan parasite Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD), between four strains of rainbow trout (Oncorhynchus mykiss) and brown trout (Salmo trutta) were evaluated. Fish were exposed to water enzootic for the parasite in the field for 5 days and were subsequently transferred to the laboratory. Relative parasite load was determined after 2, 3 and 4 weeks post-exposure (wpe) by quantitative real-time PCR (qPCR) of kidney samples and number of parasite stages was determined in immunohistochemical stained sections of kidney, liver and spleen tissues. According to qPCR results, the highest amount of parasite DNA per equal amount of host tissue at all time points was measured in brown trout. Two of the rainbow trout strains showed lower relative parasite load than all other groups at the beginning of the experiment, but the parasite multiplied faster in these strains resulting in an equal level of relative parasite load for all rainbow trout strains at 4 wpe. A weak negative correlation of fish size and parasite load was detected. Only in samples of a few fish, single stages of T. bryosalmonae were found in sections stained by immunohistochemistry impeding quantitative evaluation of parasite numbers by this method. The results indicate a differential resistance to T. bryosalmonae between the rainbow trout strains investigated and between rainbow trout and brown trout. PMID- 19683395 TI - Supraspinal role of protein kinase C in oxaliplatin-induced neuropathy in rat. AB - Oxaliplatin is a platinum-based chemotherapy drug characterized by the development of a painful peripheral neuropathy which is reproduced in rodent animal models with features observed in humans. Our focus was to explore the alterations of intracellular second messengers at supraspinal level in oxaliplatin-induced mechanical hyperalgesia. In our experiments, chronic administration of oxaliplatin to rats induced mechanical hyperalgesia which lasted for many days. When the hyperalgesic rats were submitted to paw pressure test in the presence of selective PKC inhibitor Calphostin C supraspinally administered, hyperalgesic effect could be reversed showing that PKC activity in supraspinal brain regions is needed. Concurrently, oxaliplatin chronic treatment induced a specific upregulation of gamma isoforms of PKC and increased phosphorylation of gamma/epsilon PKC isoforms within thalamus and PAG. Phosphorylation was reversed when PKC activity was inhibited by Calphostin C. Distinct PKC-activated MAPK pathways, including p38MAPK, ERK1/2 and JNK, were investigated in chronic oxaliplatin rat. A dramatic phosphorylation increase, Calphostin C sensitive, could be observed in thalamus and PAG for p38MAPK. These data show that, in oxaliplatin-induced neuropathy, enhanced mechanical nociception is strictly correlated with increased phosphorylation of specific intracellular mediators in PAG and thalamus brain regions pointing to a role of these supraspinal centers in oxaliplatin-induced neuropathic pain mechanism. PMID- 19683398 TI - Prolonged resilience of Tritrichomonas foetus in cat faeces at ambient temperature. AB - We have investigated the diagnostic utility of culture methods for Tritrichomonas foetus from cat faeces and the influence of faecal storage on the sensitivity of two different culture systems, InPouch TF-Feline (Bio-Med Diagnostics) and Modified Diamond's Medium. Normally formed cat faeces were spiked using a 10-fold serial dilution of 2 x 10(2) to 2 x 10(5)T. foetus per gram of faeces prior to storage at either room temperature or under refrigeration. At different time points a 20mg sample of spiked faeces was inoculated into each of the culture systems which were evaluated daily for up to 7 days and on day 11. Successfully cultured T. foetus 24h following faecal spiking and storage at room temperature demonstrates that the environmental resilience of the organism is greater than previously anticipated. At a conservative detection limit of 2 x 10(3) organisms per gram of faeces the accumulative sensitivity of each culture method was found to be 83% and 100% for the InPouch TF-Feline and Modified Diamond's Medium respectively over a 6h period. This research demonstrates that voided faeces kept at room temperature for up to 6h represent suitable material for diagnostic investigation. The resilience of T. foetus suggests that transmission is not limited to close contact between cats. PMID- 19683397 TI - Molecular characterization of Cryptosporidium spp. from fecal samples of birds kept in captivity in Brazil. AB - The aim of this study was to determine the prevalence of Cryptosporidium species and genotypes in birds kept in captivity in Brazil. A total of 966 samples from 18 families of birds was collected and stored in 5% potassium dichromate solution at 4 degrees C until processing. Oocysts were purified in Sheather sugar solution following extraction of genomic DNA. Molecular analyses were performed using nested-PCR for amplification of fragments of the 18S subunit of rRNA gene and of the actin gene. Amplification of Cryptosporidium DNA fragments was obtained in 47 (4.86%) samples. Sequencing of amplified fragments and phylogenetic analyses allowed the identification of Cryptosporidium baileyi in a black vulture (Coragyps atratus), a domestic chicken (Gallus gallus domesticus) and a saffron finch (Sicalis flaveola); Cryptosporidium galli in canaries (Serinus canaria), a cockatiel (Nymphicus hollandicus) and lesser seed-finches (Oryzoborus angolensis); Cryptosporidium meleagridis in a domestic chicken (G. g. domesticus); Cryptosporidium parvum in a cockatiel (N. hollandicus); Cryptosporidium avian genotype I in a canary (S. canaria) and an Indian peafowl (Pavo cristatus); Cryptosporidium avian genotype II in ostriches (Struthio camelus) and Cryptosporidium avian genotype III in a cockatiel (N. hollandicus) and a peach-faced lovebird (Agapornis roseicolis). PMID- 19683393 TI - Climatic and geographic temporal patterns of pain in the Multicenter Study of Hydroxyurea. AB - No multi-site comparisons have tested whether seasonally cold temperature or climate exacerbate pain intensity in sickle cell disease (SCD). We examined seasonal SCD pain intensity and frequency patterns and compared them with concurrent climate conditions (temperature and barometric pressure) and geography of patient residence in the Multicenter Study of Hydroxyurea (MSH). We conducted a time series analysis of the monthly average daily pain intensity (0-9 scale) and pain frequency of the 299 MSH patients from December 1991 to December 1994. We used both an unobserved component model (UCM) and a nonparametric local regression (LOESS) to probe for a cycle and/or trend associated with the time series. We also examined base mixed regression models of season, monthly average temperature and barometric pressure, and geographic region as stand-alone predictors of pain intensity and frequency. Expanded models included additional predictor variables. UCM and LOESS analyses showed a cyclic pattern of pain intensity and frequency with peaks in late Fall/early Winter and troughs in Spring. Base regression models showed colder seasons were significantly associated with greater pain intensity (p = .0035) but not frequency (p = .07); higher monthly temperatures were significantly associated with both lower pain intensity and pain frequency, but higher monthly barometric pressures were significantly associated with greater pain intensity and frequency (all p's < .0001); and northern sites had nonsignificantly higher pain intensity (p = .40) and frequency (p = .07) than southern sites. This pattern of results did not change in expanded models including other predictors. Our results suggest that seasonably colder temperatures exacerbate sickle cell-related pain, but low barometric pressure does not, and geographic region of residence is not significantly related to pain in this sample. PMID- 19683399 TI - Investigating the impact of sex and cortisol on implicit fear conditioning with fMRI. AB - Fear conditioning is influenced by stress but opposing effects in males and females have often been reported. In a previous human functional magnetic resonance imaging (fMRI) study, we observed acute effects of the stress hormone cortisol on prefrontal structures. Men showed evidence for impaired fear conditioning after cortisol treatment, while the opposite pattern was found for women. In the current experiment, we tested whether similar sex-dependent effects would occur on the neural level if contingency awareness was prevented experimentally to investigate implicit learning processes. A differential fear conditioning experiment with transcutaneous electrical stimulation as unconditioned stimulus and geometric figures as conditioned stimuli (CS) was conducted. One figure was always paired (CS+), whereas the other (CS-) was never paired with the UCS. Thirty-nine (19 female) subjects participated in this fMRI study, receiving either placebo or 30 mg cortisol (hydrocortisone) before conditioning. Dependent variables were skin conductance responses (SCRs) and neural activity (BOLD signal). In line with prior findings in unaware participants, no differential learning could be observed for the SCRs. However, a sex x cortisol interaction was detected with a reduced mean response to the CS after cortisol treatment in men, while the opposite pattern was observed in women (enhanced mean SCR under cortisol). In the contrast CS+ minus CS-, neural activity showed a sex x cortisol interaction in the insula and further trends in the hippocampus and the thalamus. In these regions, cortisol reduced the CS+/CS- differentiation in men but enhanced it in women. In contrast to these sex specific effects, differential amygdala activation was found in the placebo group but not in the cortisol group, irrespective of sex. Further, differential neural activity in the amygdala and thalamus were positively correlated with the SCRs in the placebo group only. The present study in contingency unaware participants illustrates that cortisol has in some brain regions sex specific effects on neural correlates of emotional learning. These effects might translate into a different vulnerability of the two sexes for anxiety disorders. PMID- 19683400 TI - The Substance Use Risk Profile Scale: a scale measuring traits linked to reinforcement-specific substance use profiles. AB - The Substance Use Risk Profile Scale (SURPS) is based on a model of personality risk for substance abuse in which four personality dimensions (hopelessness, anxiety sensitivity, impulsivity, and sensation seeking) are hypothesized to differentially relate to specific patterns of substance use. The current series of studies is a preliminary exploration of the psychometric properties of the SURPS in two populations (undergraduate and high school students). In study 1, an analysis of the internal structure of two versions of the SURPS shows that the abbreviated version best reflects the 4-factor structure. Concurrent, discriminant, and incremental validity of the SURPS is supported by convergent/divergent relationships between the SURPS subscales and other theoretically relevant personality and drug use criterion measures. In Study 2, the factorial structure of the SURPS is confirmed and evidence is provided for its test-retest reliability and validity with respect to measuring personality vulnerability to reinforcement-specific substance use patterns. In Study 3, the SURPS was administered in a more youthful population to test its sensitivity in identifying younger problematic drinkers. The results from the current series of studies demonstrate support for the reliability and construct validity of the SURPS, and suggest that four personality dimensions may be linked to substance related behavior through different reinforcement processes. This brief assessment tool may have important implications for clinicians and future research. PMID- 19683401 TI - Botulinum toxin-A injection may be beneficial in the treatment of life-long premature ejaculation. AB - Although premature ejaculation (PE) is a very common male sexual dysfunction there is still no European Medicines Agency (EMEA) or Food and Drug Administration (FDA) approved pharmacological treatment developed for this disease. Ejaculation is a spinal cord reflex where bulbospongiosus and ischiocavernosus muscles play the primary roles. The latest articles provided us that, life-long PE is presumed to have an underlying neurobiological pathophysiology and thus medical therapy would be superior to psychotherapy. Therefore, we hypothesize that the inhibition of the stereotyped rhythmic contractions of these muscles via the injection of botulinum toxin type A, we may have a beneficial effect in treatment of life-long PE. PMID- 19683402 TI - Upright position mechanical ventilation: an alternative strategy for ALI/ARDS patients? AB - Use of body positioning to improve oxygenation in mechanically ventilated patients with acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) has been well documented. However, neither prone position ventilation nor side lying ventilation has been reported to improve the survival. Whether there is a body position superior to routine supine position or other positions as therapeutic adjunct for ventilated patients with ALI and ARDS? We propose the hypothesis that upright position ventilation may be helpful to improve oxygenation and benefit patients with ALI/ARDS. According to the existing physiologic and pathophysiologic data of upright position investigation, we suppose that improvement of V/Q matching, increased functional residual capacity, alveolar recruitment, accelerated diaphragm recovery, early gastric emptying and enteric feeding may be a potential protect mechanism of upright position ventilation. Whether this can be translated into improvement in patient outcome should be further tested in clinical trial. PMID- 19683404 TI - Profiles of sex steroids, fecundity, and spawning of the curimata-pacu Prochilodus argenteus in the Sao Francisco River, downstream from the Tres Marias Dam, Southeastern Brazil. AB - The present study evaluated for the first time sex steroid profiles and fecundity in females of Prochilodus argenteus from two sections of the Sao Francisco River Brazil, downstream from the Tres Marias Dam, which influences characteristics of their water habitat. The model species in the study, P. argenteus, is an important commercial and recreational species in Brazil. In the region closest to the dam (section 1), females did not reach final oocyte maturation, failed to spawn, and displayed lesser circulating concentrations of testosterone, 17( hydroxyprogesterone (17(-P) and 17beta-estradiol (E2) than those farther downstream of the dam (section 2). The endocrine and fecundity deficiencies probably are attributed to lower water temperature and oxygen concentration in (section 1). The follicular atresia rate in the region closest to the dam (26%) was greater than those fish captured farther downstream of the dam (13%), after the Abaete River (section 2). Variations in testosterone, E2 and 17(-P concentrations in section 2, followed gonadal maturation which are typical features of species which have seasonal reproduction, group-synchronous oocyte development, and are single batch spawners such as P. argenteus. Results document the first evidence of endocrine and reproductive dysfunctions caused by inadequate water conditions in a wild population of the migratory species P. argenteus in the Sao Francisco River, downstream from the Tres Marias dam. PMID- 19683403 TI - Automated radiation targeting in head-and-neck cancer using region-based texture analysis of PET and CT images. AB - PURPOSE: A co-registered multimodality pattern analysis segmentation system (COMPASS) was developed to automatically delineate the radiation targets in head and-neck cancer (HNC) using both (18)F-fluoro-deoxy glucose-positron emission tomography (PET) and computed tomography (CT) images. The performance of the COMPASS was compared with the results of existing threshold-based methods and radiation oncologist-drawn contours. METHODS AND MATERIALS: The COMPASS extracted texture features from corresponding PET and CT voxels. Using these texture features, a decision-tree-based K-nearest-neighbor classifier labeled each voxel as either "normal" or "abnormal." The COMPASS was applied to the PET/CT images of 10 HNC patients. Automated segmentation results were validated against the manual segmentations of three radiation oncologists using the volume, sensitivity, and specificity. The performance of the COMPASS was compared with three PET-based threshold methods: standard uptake value of 2.5, 50% maximal intensity, and signal/background ratio. RESULTS: The tumor delineations of the COMPASS were both quantitatively and qualitatively more similar to those of the radiation oncologists than the delineations from the other methods. The specificity was 95% +/- 2%, 84% +/- 9%, 98% +/- 3%, and 96% +/- 4%, and the sensitivity was 90% +/- 12%, 93% +/- 10%, 48% +/- 20%, and 68% +/- 25% for the COMPASS, for a standard uptake value of 2.5, 50% maximal intensity, and signal/background ratio, respectively. The COMPASS distinguished HNC from adjacent normal tissues with high physiologic uptake and consistently defined tumors with large variability in (18)F-fluoro-deoxy glucose uptake, which are often problematic with the threshold based methods. CONCLUSION: Automated segmentation using texture analysis of PET/CT images has the potential to provide accurate delineation of HNC. This could lead to reduced interobserver variability, reduced uncertainty in target delineation, and improved treatment planning accuracy. PMID- 19683405 TI - [Estimating the incidence of colorectal cancer in France from a hospital discharge database, 1999-2003]. AB - BACKGROUND: Cancer incidence in France is monitored by district-level cancer registries, which cover only 15% of the population. Incidence at the national and regional level are estimated from mortality data by extrapolating the ratio between incidence and mortality observed in the districts covered by a cancer registry. Using the incidence/mortality ratio might not be relevant at the district-level (departement). This study aims to produce district-level estimations of colorectal cancer incidence, using the ratio between incident cases from cancer registries and surgery admissions for colorectal cancer identified in the national hospital discharge database. METHODS: This ratio was studied for the period 1999-2003 in the 13 districts covered by a cancer registry. For each sex separately, the number of incident cases was analyzed according to the number of surgery admissions for resection of colorectal cancer using a Poisson model. Age was introduced in the model as a fixed effect and district as a random effect. The model's ability to predict incidence was tested through cross-validation. The model was then extrapolated in order to estimate incidence of colorectal cancer in all French districts. RESULTS: In the districts covered by a cancer registry, cross-validation showed the model had a good predictive ability, except in men for one district where the difference between predicted and observed incidence reached 10%. Estimated incidence rates, age standardized on the world population, ranged broadly from 29 to 44 per 100,000 in men and from 17 to 27 per 100,000 in women. Incidence did not show any clear geographical pattern. CONCLUSION: Among districts covered by a cancer registry, cross-validation showed overall good accuracy of predicted incidence. Inclusion of several admissions per patient was certainly a minor source of error in these estimations. Indeed, our selection only included 2% of multiple admissions, without geographical variations, in 2002 and 2003, years for which patient identifiers were available in the hospital discharge database. Estimated incidence rates presented moderate geographical variations and their prediction intervals should be taken into account. PMID- 19683406 TI - [The future of gastrointestinal therapeutic endoscopy: NOTES]. AB - Natural orifice translumenal endoscopic surgery (NOTES) allows access into the peritoneal cavity with a flexible endoscope, through the wall of the digestive or urogenital tracts. NOTES can be combined to laparoscopic surgery in so-called << hybrid >> techniques. In the absence of any incision of the abdominal wall, NOTES procedures provide perfect cosmetic results, with virtually no risk of parietal complications, and with decreased postoperative pain. NOTES could particularly benefit to overweight patients and to patients receiving intensive or palliative care. Most NOTES studies have been performed on animal models, with great interest for both transgastric and transpelvic approaches. Successful NOTES peritoneoscopy, hysterectomy, oophorectomy, tubal ligation, gastrojejunal anastomosis, cholecystectomy, splenectomy, nephrectomy, and abdominal-wall hernia repair have been described. In human studies, the transvaginal route is preferred. NOTES clinical research focuses on low-morbidity procedures, such as cholecystectomy, appendectomy, and peritoneoscopy. Indirect benefits are expected from this research, with possible technological innovations in the field of endoscopic instrumentation (including sutures, anastomosis, traction and triangulation). Overall, NOTES is believed to make evolve both interventional endoscopy and minimally invasive surgery. PMID- 19683407 TI - Patient-doctor interaction in rehabilitation: the relationship between perceived interaction quality and long-term treatment results. AB - OBJECTIVES: A body of evidence suggests that good interaction is crucial for high quality medical practice and has a considerable impact on treatment outcomes. Less is known about the role and significance of doctor-patient interaction in rehabilitation. The study aim was to capture perceived quality of doctor-patient interaction in rehabilitation by a rating instrument (P.A.INT-Questionnaire. P.A.INT is the abbreviation for Patient-Arzt-Interaktion (German)) and to examine the relationship between perceived quality of interaction and long-term treatment outcomes. METHODS: Referring to the approach of Bensing [Bensing JM. Doctor patient communication and the quality of care. Utrecht: NIVEL; 1990] we defined "quality of interaction" in terms of three dimensions: (1) affective behaviour, i.e. empathy, positive regard and coherence [Rogers CR. Die nicht direktive Beratung Munchen: Kindler Studienausgabe [Counselling and psychotherapy, 1942]. Boston; 1972]; (2) instrumental behaviour: providing and collecting information, structuring and reinforcement; (3) participation and involvement of patients. Two parallel versions of the questionnaire were developed for patients and physicians. Seven rehabilitation clinics in north western Germany participated in the multi-centre study. Sixty-one doctors and their four hundred and seventy patients evaluated both their shared dialogues upon admission, discharge and ward round. Furthermore, patients rated their health status on admission (t0), discharge (t1) and six months after discharge (t2) with the IRES-3 (Indicators of Rehabilitation Status Questionnaire, Version 3). RESULTS: (1) Comparisons of patient and physician evaluations on admission revealed the following: affective quality of contact (empathy and coherence) was rated positively and without discrepancies by both patients and physicians. On the other hand, instrumental behaviour (information and structuring) was rated less positively by patients than by physicians. (2) Patients who rated the dialogue on admission more positively showed stronger treatment effects with respect to pain as well as to anxiety at discharge and six months after discharge. Analysis for single scales of the P.A.INT-Questionnaire revealed that this is due to affective and instrumental quality of the dialogues. CONCLUSION: Our results suggest a correlation between perceived interaction quality, as defined by our questionnaire and treatment effects six months after discharge. Comparisons of patient and physician evaluations showed that physicians seem to be successful in building relationships on the affective level, but less successful on the instrumental level (i.e. information, structuring and reinforcement). They also perceive disturbances on the relational and organisational level more strongly than patients. PRACTICE IMPLICATIONS: Our data underline the importance of interaction quality for the success of rehabilitation and thus the importance of specific skills such as providing and collecting information, recognizing patients' concerns and goals as well as reinforcement of health related action. Regular training and supervision should be provided to support physicians and to enhance their competence in dealing with patients concerns. PMID- 19683408 TI - Disclosing medical errors to patients: a challenge for health care professionals and institutions. AB - Medical errors occur frequently in routine health care. Historically, many of these errors were not disclosed to patients but increasingly there are calls for frank and open disclosure of errors to patients and families. This article provides an overview of what information should be disclosed about errors, what patients want to be told, the attitudes and skills of physicians in disclosure, and the barriers to effective disclosure. This article also includes a description of the changing policy environment in North America that is encouraging and mandating disclosure of errors. PMID- 19683409 TI - Chicken folliculo-stellate cells express thyrotropin receptor mRNA. AB - We investigated the presence of thyrotropin receptor (TSHR) mRNA in chicken pituitary and brain, and quantified the changes in its expression during the last week of embryonic development. We found that in the pituitary gland, TSHR mRNA co localizes with folliculo-stellate cells but not with thyrotropic cells, suggesting the existence of a paracrine ultra-short thyrotropin feedback loop. TSHR mRNA was also present throughout the diencephalon and various other brain regions, which implies a more general function for thyrotropin in the avian brain. During late embryogenesis, when the activity of the hypothalamo-pituitary thyroidal axis increases markedly, a significant rise in TSHR mRNA expression was observed in pituitary, which may signify an important change in pituitary ultra short thyrotropin feedback regulation around the period of hatching. PMID- 19683410 TI - [Necrosis of the round liver ligament: a rare cause of acute abdominal pain]. PMID- 19683411 TI - [The future of electronic biomedical journals]. AB - Old journals are using information technologies by putting production pdfs on line and applying several useful functions, such as search engines or podcasts. Journals have rapidly taken up the Electronic Long Paper Short (ELPS) trend, publishing abridged articles in the physical journal and longer versions on their websites, with electronic supplements (supplementary references, spreadsheets, videos, etc.). Journals with no paper version have been created that use all the functionalities of the Internet and now of web 2.0. They are numerous (PLoS, BioMed Primary, etc..) and are acquiring a reputation. Free access for all, information for all in 2015, are some interesting objectives that remain to be funded. The economic model of these exclusively electronic journals is uncertain, although some are funded by authors (and not at all by readers) or institutions. Spotless research is a rare exception, but few stakeholders are taking an active interest in it. Humans, much more than the tool or the media, are at the heart of the system. We must remember that those who have wanted to predict the future have always been wrong. PMID- 19683412 TI - Network mechanisms of gamma oscillations in the CA3 region of the hippocampus. AB - Neural networks of the brain display multiple patterns of oscillatory activity. Some of these rhythms are generated intrinsically within the local network, and can therefore be studied in isolated preparations. Here we discuss local-circuit mechanisms involved in hippocampal CA3 gamma oscillations, one of the best understood locally generated network patterns in the mammalian brain. Perisomatic inhibitory cells are crucial players in gamma oscillogenesis. They provide prominent rhythmic inhibition to CA3 pyramidal cells and are themselves synchronized primarily by excitatory synaptic inputs derived from the local collaterals of CA3 pyramidal cells. The recruitment of this recurrent excitatory inhibitory feedback loop during hippocampal gamma oscillations suggests that local gamma oscillations not only control when, but also how many and which pyramidal cells will fire during each gamma cycle. PMID- 19683413 TI - Systematic review data extraction: cross-sectional study showed that experience did not increase accuracy. AB - OBJECTIVE: This study assessed the impact of systematic review and data extraction experience on the accuracy and efficiency of data extraction in systematic reviews. STUDY DESIGN AND SETTING: We conducted a prospective cross sectional study from October to December 2006. Participants were classified as having minimal, moderate, or substantial experience in systematic reviews and data extraction. Three studies on insomnia treatment were extracted. Our primary outcome was the accuracy of data extraction. Data sets of each experience level were analyzed for errors in data extraction and results of meta-analyses. Additionally, the time required for completion of data extraction was compared. RESULTS: Error rates were similar across the various levels of experience and ranged from 28.3% to 31.2%. Mean rates for errors of omission (11.3-16.4%) were generally lower than those for errors of inaccuracy (13.9-17.9%). There were no significant differences in error rates or accuracy of meta-analysis results between groups. Time required approached significance, with minimally experienced participants requiring the most time. CONCLUSION: Overall, there were high error rates by participants at all experience levels; however, time required for extraction tended to decrease with experience. These results illustrate the need to develop strategies aimed at mastery of data extraction, rather than reliance on previous data extraction experience alone. PMID- 19683414 TI - Shoulder-specific disability measures showed acceptable construct validity and responsiveness. AB - OBJECTIVE: To compare the construct validity and responsiveness of the Shoulder Pain and Disability Index (SPADI), Croft Index, and disabilities of the arm, shoulder, and hand (DASH) for patients with adhesive capsulitis and to compare these with pain on a visual analog scale, the Health Assessment Questionnaire, and the problem-elicitation technique. STUDY DESIGN AND SETTING: Two randomized double-blind placebo-controlled trials of interventions for adhesive capsulitis were performed. Both trials recruited patients from community-based physiotherapy practices. Responsiveness at 3 weeks postbaseline was assessed using four responsiveness parameters and three external criteria for improvement. RESULTS: Correlations between the shoulder-specific measures ranged from 0.55 to 0.65 at baseline and 0.49-0.55 for the 3-week change scores. Greater responsiveness was seen for the SPADI. The effect size for the SPADI ranged from 1.20 to 1.64, for the Croft Index from 0.87 to 1.21, and for the DASH from 0.55 to 0.83. Rankings were similar across the four responsiveness parameters and the three external criteria for improvement. Correlations between the shoulder-specific and generic measures for baseline and 3-week change scores were lower than those among the shoulder-specific measures (range: 0.17-0.60). CONCLUSION: The shoulder-specific disability measures showed acceptable construct validity and responsiveness with a small but consistent overall advantage for the SPADI. PMID- 19683415 TI - SLA/LP/tRNP((Ser)Sec) antigen in autoimmune hepatitis: identification of the native protein in human hepatic cell extract. AB - A diagnostic subgroup of AIH type 1 is characterized by specific serum antibodies against soluble liver protein. The respective autoantigen was named SLA/LP/tRNP((Ser)Sec), after three homologous recombinant polypeptides were isolated from expression gene libraries. We analyzed human cultured liver cells for the human homologue of recombinant SLA/LP/tRNP((Ser)Sec) by antigen purification. In addition, a monoclonal antibody was generated against recombinant SLA-p35, a truncated recombinant SLA-reactive polypeptide. With a positive patient serum, immune affinity chromatography was performed on the 52 kD SLA main antigenic determinant pre-enriched by ion exchange chromatography. By mass spectrometry, the 52 kD-SLA/LP/tRNP ((Ser)Sec) autoantigen was unambiguously identified in the purification product. The identity of the recombinant SLA-p35 and its human homologue was further confirmed by a specific signal of the anti SLA-p35 monoclonal antibody with purified human SLA/LP/tRNP((Ser)Sec). The 48 kD SLA species frequently comigrating in SLA-immunoblotting however was not identified by either approach. We conclude that the native counterpart of recombinant tRNP((Ser)(Sec)) indeed is detectable with a molecular weight of 52 kD in soluble liver extract of human cells as the major antigenic component of SLA/LP/tRNP((Ser)Sec). PMID- 19683416 TI - Synovial chondromatosis of the temporomandibular joint accompanied by loose bodies in both the superior and inferior joint compartments: case report. AB - Synovial chondromatosis (SC) of the temporomandibular joint (TMJ) is a benign lesion characterized by the formation of metaplastic cartilaginous nodules. SC of the TMJ usually only affects the superior joint compartment of the TMJ. The authors report a rare case of SC of the TMJ affecting the inferior and superior joint compartments. PMID- 19683417 TI - Paternal age as a risk factor for schizophrenia: how important is it? AB - Advanced paternal age has been widely cited as a risk factor for schizophrenia among offspring and even claimed to account for one-quarter of all cases. We carried out a new study on 25,025 offspring from the Collaborative Perinatal Project (CPP), including 168 diagnosed with psychosis and 88 with narrowly defined schizophrenia. We also conducted a meta-analysis of this and nine other studies for which comparable age-cohort data were available. The mean paternal age for the CPP cases was slightly, but not significantly, higher than the matched controls (p=0.28). Meta-analyses including these new results were conducted to determine the relative risk associated with alternative definitions of advanced paternal age (35, 45 or 55 years and older). These yielded pooled odds ratios and 95% confidence intervals of 1.28 (1.10, 1.48), 1.38 (0.95, 2.01) and 2.22 (1.46, 3.37), respectively. Thus, increased paternal age appears to be a risk factor for schizophrenia primarily among offspring of fathers ages 55 and over. In these 10 studies, such fathers accounted for only 0.6% of all births. Compared with other known risk factors for schizophrenia, advanced paternal age appears to be intermediate in magnitude. Advanced paternal age is also known to be a risk factor for some chromosomal and neoplastic diseases in the offspring where the cause is thought to be chromosomal aberrations and mutations of the aging germline. Similar mechanisms may account for the relationship between advanced paternal age and schizophrenia risk. PMID- 19683418 TI - Progressive gray matter changes in first episode schizophrenia: a 4-year longitudinal magnetic resonance study using VBM. AB - Schizophrenia is a disabling illness, characterized by a heterogeneous course including clinical deterioration and poor outcome. Accumulating findings in schizophrenia suggest that it might involve two pathophysiologic processes, one early in life (neurodevelopmental), and one after onset of the illness (neurodegenerative). Longitudinal imaging studies after onset of the illness may help to clarify these pathophysiological aspects of schizophrenia, but so far, probably due to methodological differences, there have been no conclusive results. The present study sets out to investigate longitudinal gray matter changes in patients with first-episode schizophrenia relative to healthy subjects over the first 4 years of the illness and the relation of gray matter changes in patients with functional outcome, using an objective automatic method not biased to one particular structure to analyze gray matter changes. METHODS: We included 28 first-episode neuroleptic-naive patients with DSM-IV diagnosis of schizophreniform disorder or schizophrenia, and 17 controls. 15 patients and 11 controls completed the longitudinal study and were reevaluated after four years. Gray matter changes over time were measured with voxel-based morphometry (VBM) using SPM5. Functional outcome was measured with the global assessment functioning scale (GAF). RESULTS: Excessive decrease in gray matter was found in patients as compared to healthy individuals in the left superior temporal gyrus and right orbitofrontal gyrus, and excessive increase in the bilateral lingual gyrus and right cuneus. Additionally, gray matter changes in patients in the left lingual gyrus, right insula and right cerebellum, were inversely related to functional outcome (p<0.001 uncorrected at voxel level, p<0.05 family-wise-error corrected at cluster level). CONCLUSIONS: There are differing longitudinal gray matter changes in patients with schizophrenia during the first years of the illness as compared to healthy individuals. Some progressive gray matter changes in patients are related to functional outcome. PMID- 19683419 TI - Patterns of brain activity during a semantic task differentiate normal aging from early Alzheimer's disease. AB - In a study of the effects of normal and pathological aging on semantic-related brain activity, 29 patients with Alzheimer's disease (AD) and 19 controls subjects (10 young and 9 older controls) performed a version of the Pyramids and Palm Trees Test that had been adapted for use during functional magnetic resonance imaging (fMRI). Young and older controls activated the left inferior and middle frontal gyri, precuneus and superior parietal lobule. Right frontal and left temporal cortices were activated only in the young. The AD group activated only the left prefrontal and cingulate cortex. Separate analyses of high- and low-performing AD subgroups showed a similar pattern of activation in the left frontal lobe, although activiation was more widespread in low performers. High performers significantly deactivated anterior midline frontal structures, however, while low performers did not. When the older adult and AD groups were combined, there was a significant positive correlation between left frontal and parietal activation and Mini-Mental State Examination (MMSE) score (covarying for age), suggesting a disease effect. A significant negative correlation between activation in the left temporal cortex and age (covarying for MMSE score) reflected a possible age effect. These differential effects suggest that semantic activation paradigms might aid diagnosis in those cases for whom conventional assessments lack the necessary sensitivity to detect subtle changes. PMID- 19683420 TI - [Congenital chylothorax and hypothyroidism: a case report and a review of the literature]. AB - Congenital chylothorax, an uncommon disorder, is a therapeutic challenge without satisfactory results. When classical medical approaches fail (such as thoracosynthesis, total parenteral nutrition, and fasting followed by oral medium chain triglycerides), some medical teams introduce somatostatin or octreotide in cases of recurring chylothorax. We report a case of recurring chylothorax treated with somatostatin at day 27 correlated with an unfortunate discovery of hypothyroidism on day 34. Clinical signs of chylothorax on somatostatin clearly improved with the introduction of levothyroxine. This article points out the relation between the two diseases based on a review of the literature. PMID- 19683421 TI - Antimycobacterial activity of lichen substances. AB - We describe here the extraction and identification of several classes of phenolic compounds from the lichens Parmotrema dilatatum (Vain.) Hale, Parmotrema tinctorum (Nyl.) Hale, Pseudoparmelia sphaerospora (Nyl.) Hale and Usnea subcavata (Motyka) and determined their anti-tubercular activity. The depsides (atranorin, diffractaic and lecanoric acids), depsidones (protocetraric, salazinic, hypostictic and norstictic acids), xanthones (lichexanthone and secalonic acid), and usnic acid, as well seven orsellinic acid esters, five salazinic acid 8',9'-O-alkyl derivatives and four lichexanthone derivatives, were evaluated for their activity against Mycobacterium tuberculosis. Diffractaic acid was the most active compound (MIC value 15.6mug/ml, 41.6 microM), followed by norstictic acid (MIC value 62.5 microg/ml, 168 microM) and usnic acid (MIC value 62.5 microg/ml, 182 microM). Hypostictic acid (MIC value 94.0 microg/ml, 251 microM) and protocetraric acid (MIC value 125 microg/ml, 334 microM) showed moderate inhibitory activity. The other compounds showed lower inhibitory activity on the growth of M. tuberculosis, varying from MIC values of 250 to 1370 microM. PMID- 19683423 TI - DNA detection using organic thin film transistors: optimization of DNA immobilization and sensor sensitivity. AB - Advancements in research and technology are happening now more than ever in the biotechnology industry. Better detection and treatment methods, specifically targeted towards personalized systems, are constantly being sought by researchers. Collaboration among different fields of science and technology has brought us one step closer towards achieving this goal. Recently, the use of pentacene thin film transistors as DNA hybridization detection systems has been demonstrated, leading to the possibility of their use in the implementation of electrically read low-cost DNA microarrays. Physical adsorptive immobilization, the mechanism through which DNA immobilizes on pentacene, highlights the importance of the topology of the pentacene film surface for immobilization of DNA and the sensitivity of the sensor. DNA is known to segregate to topological features on pentacene surface. We are able to exploit the control of pentacene evaporation conditions to tune pentacene film morphology to maximize sensitivity. In this paper, we demonstrate and analyze DNA detection using optimized films. PMID- 19683422 TI - The general utilization of scrapped PC board. AB - The traditional burning process is used to recover copper from scrapped PC board (printed circuit board) but it causes serious environmental problems. In this research a new process was developed which not only prevents pollution problems, but also maximizes the utility of all the materials on the waste board. First, the scrapped PC board was crushed and grounded, then placed in the NH3/NH5CO3 solution with aeration in order to dissolve copper. After distilling the copper NH3/NH5CO3 solution and then heating the distilled residue of copper carbonate, pure copper oxide was obtained with particle size of about 0.2 microm and the shape elliptical. The remaining solid residue after copper removal was then leached with 6N hydrochloric acid to remove tin and lead. The last residue was used as a filler in PVC plastics. The PVC plastics with PC board powder as filling material was found to have the same tensile strength as unfilled plastics, but had higher elastic modulus, higher abrasion resistance, and was cheaper. PMID- 19683424 TI - A glucose biosensor based on direct electrochemistry of glucose oxidase immobilized on nitrogen-doped carbon nanotubes. AB - A novel biosensor for glucose was prepared by immobilizing glucose oxidase (GOx) on nitrogen-doped carbon nanotubes (CNx-MWNTs) modified electrode. The CNx-MWNTs membrane showed an excellent electrocatalytic activity toward the reduction of O(2) due to its diatomic side-on adsorption on CNx-MWNTs. The nitrogen doping accelerated the electron transfer from electrode surface to the immobilized GOx, leading to the direct electrochemistry of GOx. The biofunctional surface showed good biocompatibility, excellent electron-conductive network and large surface-to volume ratio, which were characterized by scanning electron microscopy, contact angle and electrochemical impedance technique. The direct electron transfer of immobilized GOx led to stable amperometric biosensing for glucose with a linear range from 0.02 to 1.02 mM and a detection limit of 0.01 mM (S/N=3). These results indicated that CNx-MWNTs are good candidate material for construction of the third-generation enzyme biosensors based on the direct electrochemistry of immobilized enzymes. PMID- 19683425 TI - Nanoparticle carrying a single probe for target DNA detection and single nucleotide discrimination. AB - The identification of single nucleotide mutations with specific disease and single nucleotide polymorphisms (SNPs) among individuals is increasingly important for diagnosis of genetic disease, prediction of disease resistance or predispositions, as well as administration of drug dosages and design of personalized medicine. In this study, we demonstrated a convenient yet useful colorimetric quantitative DNA assay method with high single nucleotide discrimination for both center and end-mismatched sequences. The detection limit of our method is 75 fmol of DNA sample. Even for mixed DNA sample with low percentages of matched targets, this method shows good probe selectivity and zero false positive detection. Finally, the ease of operation and compatibility with existing molecular biology toolbox makes this method a potential low-cost alternative in scientific and clinical diagnostic application. PMID- 19683426 TI - A cytokine immunosensor for Multiple Sclerosis detection based upon label-free electrochemical impedance spectroscopy using electroplated printed circuit board electrodes. AB - A biosensor for the serum cytokine, Interleukin-12 (IL-12), based upon a label free electrochemical impedance spectroscopy (EIS) monitoring approach is described. Overexpression of IL-12 has been correlated to the diagnosis of Multiple Sclerosis (MS). An immunosensor has been fabricated by electroplating gold onto a disposable printed circuit board (PCB) electrode and immobilizing anti-IL-12 monoclonal antibodies (MAb) onto the surface of the electrode. This approach yields a robust sensor that facilitates reproducible mass fabrication and easy alteration of the electrode shape. Results indicate that this novel PCB sensor can detect IL-12 at physiological levels, <100 fM with f-values of 0.05 (typically <0.0001) in a label-free and rapid manner. A linear (with respect to log concentration) detectable range was achieved. Detection in a complex biological solution is also explored; however, significant loss of dynamic range is noted in the 100% complex solution. The cost effective approach described here can be used potentially for diagnosis of diseases (like MS) with known biomarkers in body fluids and for monitoring physiological levels of biomolecules with healthcare, food, and environmental relevance. PMID- 19683427 TI - Engineering and design: editorial overview. PMID- 19683428 TI - Anxiety and borderline PAP smear results. AB - PURPOSE: Low-grade abnormalities after cervical cancer screening, i.e. borderline (Pap 2) or mildly (Pap 3a1) dyskaryotic (BMD) smear results, are found in considerable numbers of women annually. We compared quality of life and anxiety in women with BMD and a reference group of screening participants who were awaiting smear taking. METHODS: Five hundred and fifty women with BMD in the preceding 6-24 months, identified through a regional screening organisation, were sent a questionnaire addressing generic quality of life (12-item Short-Form Health Survey [SF-12], EuroQol classification [EQ-5D]), generic anxiety (STAI-6) and screen-specific anxiety (Psychological Consequences Questionnaire [PCQ]). RESULTS: After adjustment for differences in background characteristics, women with BMD (n=270) reported more generic anxiety (44.4 versus 32.6) and screen specific anxiety (5.0 versus 1.4) than the reference group (n=352). The differences indicated statistical (p<0.001) and clinical significance. High anxiety (STAI-6>44) was reported by 49% of the BMD group. Mental quality of life was worse in the BMD than in the reference group (44.2 versus 52.0, p<0.001). The BMD group considered screening more often frightening (27% versus 10%) and reported 'fear for cervical cancer' more frequently as their reason for having a (repeat) smear taken (62/270, 23% versus 12/346, 4%). CONCLUSION/DISCUSSION: BMD smears were consistently associated with considerable excess anxiety in the period of 6-24 months after the original BMD Pap smears had been taken. PMID- 19683430 TI - Functional activity of CXCL8 receptors, CXCR1 and CXCR2, on human malignant melanoma progression. AB - We examined the autocrine/paracrine role of interleukin-8 (CXCL8) and the functional significance of CXCL8 receptors, CXCR1 and CXCR2, in human malignant melanoma proliferation, migration, invasion and angiogenesis. We found that a panel of seven cell lines, even though at different extent, secreted CXCL8 protein, and expressed CXCR1 and CXCR2 independently from the CXCL8 expression, but depending on the oxygen level. In fact, hypoxic exposure increases the expression of CXCR1 and CXCR2. The cell proliferation of both M20 and A375SM lines, expressing similar levels of both CXCR1 and CXCR2 but secreting low and high amounts of CXCL8, respectively, was significantly enhanced by CXCL8 exposure and reduced by CXCL8, CXCR1 and CXCR2 neutralising antibodies, indicating the autocrine/paracrine role of CXCL8 in melanoma cell proliferation. Moreover, an increased invasion and migration in response to CXCL8 was observed in several cell lines, and a further enhancement evidenced under hypoxic conditions. A CXCL8 dependent in vivo vessel formation, evaluated through a matrigel assay, was also demonstrated. Furthermore, when neutralising antibodies against CXCR1 or CXCR2 were used, only the involvement of CXCR2, but not CXCR1 was observed on cell migration and invasion, while both receptors played a role in angiogenesis. In summary, our data demonstrate that CXCL8 induces cell proliferation and angiogenesis through both receptors and that CXCR2 plays an important role in regulating the CXCL8-mediated invasive and migratory behaviour of human melanoma cells. Thus, blocking the CXCL8 signalling axis promises an improvement for the therapy of cancer and, in particular, of metastatic melanoma. PMID- 19683429 TI - Activity of ixabepilone in oestrogen receptor-negative and oestrogen receptor progesterone receptor-human epidermal growth factor receptor 2-negative metastatic breast cancer. AB - Oestrogen receptor (ER)-negative breast cancer, including oestrogen receptor-, progesterone receptor- and human epidermal growth factor receptor 2-negative (ER/PR/HER2-negative) breast cancer, is more aggressive than ER-positive disease. A major limitation in the treatment of ER-negative disease subtypes is the inherent insensitivity to hormonal agents (tamoxifen, aromatase inhibitors) that are widely used in the treatment of breast cancer. Thus, therapeutic options for poor prognosis patients with ER-negative breast cancer are limited to a handful of chemotherapeutic agents, and new agents are needed to improve the treatment of this disease. Ixabepilone, a novel epothilone B analogue with low susceptibility to cellular mechanisms that confer resistance to taxanes and other chemotherapeutic agents, has demonstrated potent preclinical antitumour activity in multiple models, including those with primary or acquired drug resistance. This review summarises the results of a prospective subset analysis from a phase III clinical trial evaluating ixabepilone for the treatment of metastatic breast cancer (MBC), in which efficacy and safety were evaluated in patients with ER negative and ER/PR/HER2-negative disease. PMID- 19683431 TI - Cardiorespiratory fitness, lifestyle factors and cancer risk and mortality in Finnish men. AB - BACKGROUND: Physical fitness along with lifestyle factors may have important roles in the prevention of cancer. We examined the relationship between common lifestyle factors such as energy expenditure, physical activity and maximal oxygen uptake (VO(2max)), nutrition and smoking habits and the risk of cancer. METHODS: A population-based cohort study was carried out in 2268 men from Eastern Finland with no history of cancer. They were followed up for an average of 16.7 years. The outcome measures were cancer incidence (n=387) and cancer mortality (n=159). RESULTS: Men with VO(2max) of more than 33.2 mL/kg/min (highest tertile) had 27% (95% confidence interval (CI) 0.56-0.97) decreased cancer incidence and 37% (95% CI 0.40-0.97) reduced cancer mortality than men with VO(2max) of less than 26.9 mL/kg/min (lowest tertile) after adjustment for age, examination year, alcohol, smoking, socioeconomic status, waist-to-hip ratio and energy, fibre and fat intake. The risk reduction was mainly due to decreased risk of lung cancer in fit men. The adjusted risk of cancer was 0.73 (95% CI 0.55-0.98) among fit (VO(2max)> or =26.9 mL/kg/min) men with the total energy expenditure of physical activity over 2500 kcal/week. A total of 290 active (energy expenditure >2500 kcal and at least 2h of physical activity per week) men with a favourable lifestyle (good fitness, balanced diet and non-smoking) had an adjusted relative risk of 0.63 (95% CI 0.46-0.87) for cancer. CONCLUSION: Favourable lifestyle including good cardiorespiratory fitness and healthy dietary habits with active and non-smoking lifestyle considerably reduces the risk of cancer. PMID- 19683433 TI - Biocompatible epoxy modified bio-based polyurethane nanocomposites: mechanical property, cytotoxicity and biodegradation. AB - Epoxy modified Mesua ferrea L. seed oil (MFLSO) based polyurethane nanocomposites with different weight % of clay loadings (1%, 2.5% and 5%) have been evaluated as biocompatible materials. The nanocomposites were prepared by ex situ solution technique under high mechanical shearing and ultrasonication at room temperature. The partially exfoliated nanocomposites were characterized by Fourier transform infra-red (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM) and transmission electron microscopy (TEM) techniques. The mechanical properties such as tensile strength and scratch hardness were improved 2 and 5 times, respectively by nanocomposites formation. Even the impact resistance improved a little. The thermostability of the nanocomposites was enhanced by about 40 degrees C. Biodegradation study confirmed 5-10 fold increase in biodegradation rate for the nanocomposites compared to the pristine polymers. All the nanocomposites showed non-cytotoxicity as evident from RBC hemolysis inhibition observed in anti-hemolytic assay carried over the sterilized films. The study reveals that the epoxy modified MFLSO based polyurethane nanocomposites deserve the potential to be applicable as biomaterials. PMID- 19683432 TI - What determines individuals' preferences for colorectal cancer screening programmes? A discrete choice experiment. AB - INTRODUCTION: In many countries uptake of colorectal cancer (CRC) screening remains low. AIM: To assess how procedural characteristics of CRC screening programmes determine preferences for participation and how individuals weigh these against the perceived benefits from participation in CRC screening. METHODS: A discrete choice experiment was conducted among subjects in the age group of 50-75 years, including both screening-naive subjects and participants of a CRC screening programme. Subjects were asked on their preferences for aspects of CRC screening programmes using scenarios based on pain, risk of complications, screening location, preparation, duration of procedure, screening interval and risk reduction of CRC-related death. RESULTS: The response was 31% (156/500) for screening-naive and 57% (124/210) for CRC screening participants. All aspects proved to significantly influence the respondents' preferences. For both groups combined, respondents required an additional relative risk reduction of CRC related death by a screening programme of 1% for every additional 10 min of duration, 5% in order to expose themselves to a small risk of complications, 10% to accept mild pain, 10% to undergo preparation with an enema, 12% to use 0.75l of oral preparation combined with 12h fasting and 32% to use an extensive bowel preparation. Screening intervals shorter than 10 years were significantly preferred to a 10-year screening interval. CONCLUSION: This study shows that especially type of bowel preparation, risk reduction of CRC related death and length of screening interval influence CRC screening preferences. Furthermore, improving awareness on CRC mortality reduction by CRC screening may increase uptake. PMID- 19683434 TI - Laccase detoxification of steam-exploded wheat straw for second generation bioethanol. AB - In this work we compared the efficiency of a laccase treatment performed on steam exploded wheat straw pretreated under soft conditions (water impregnation) or harsh conditions (impregnation with diluted acid). The effect of several enzymatic treatment parameters (pH, time of incubation, laccase origin and loading) was analysed. The results obtained indicated that severity conditions applied during steam explosion have an influence on the efficiency of detoxification. A reduction of the toxic effect of phenolic compounds by laccase polymerization of free phenols was demonstrated. Laccase treatment of steam exploded wheat straw reduced sugar recovery after enzymatic hydrolysis, and it should be better performed after hydrolysis with cellulases. The fermentability of hydrolysates was greatly improved by the laccase treatment in all the samples. Our results demonstrate the action of phenolic compounds as fermentation inhibitors, and the advantages of a laccase treatment to increase the ethanol production from steam-exploded wheat straw. PMID- 19683435 TI - Anaerobic degradation pathway and kinetics of domestic wastewater at low temperatures. AB - The effect of temperatures below 20 degrees C (20, 15 and 10 degrees C) on the anaerobic degradation pathway and kinetics of domestic wastewater fractionated at different sizes was studied in a fluidized-bed batch reactor. The overall degradation pathway was characterized by a soluble fraction degrading according to zero-order kinetics and a colloidal fraction (between 0.45 and 4.5 microm) that first disintegrates into a particulate fraction smaller than 0.45 microm before finally degrading. The colloidal degradation processes follow a first order kinetic. In contrast, suspended solids (bigger than 4.5 microm) degrade to soluble and colloidal fractions according to first-order kinetics. The colloidal fraction originating from suspended solids further degrades into soluble fraction. These soluble fractions have the same degradation kinetics as the original soluble fraction. The suspended solids degradation was highly affected by temperature, whereas the soluble fraction slightly affected and the colloidal fraction was not affected at all. On the other hand, the colloidal non-degradable fraction increased significantly with the decrease in temperature while the suspended solids slowly increased. The soluble non-degradable fraction was little affected by temperatures changes. PMID- 19683436 TI - Chemical composition and antibacterial activity of Cordia verbenacea extracts obtained by different methods. AB - The present study describes the chemical composition and the antibacterial activity of extracts from Cordia verbenacea DC (Borraginaceae), a traditional medicinal plant that grows widely along the southeastern coast of Brazil. The extracts were obtained using different extraction techniques: high-pressure operations and low-pressure methods. The high-pressure technique was applied to obtain C. verbenacea extracts using pure CO(2) and CO(2) with co-solvent at pressures up to 30MPa and temperatures of 30, 40 and 50 degrees C. Organic solvents such as n-hexane, ethyl acetate, ethanol, acetone and dichloromethane were used to obtain extracts by low-pressure processes. The antibacterial activity of the extracts was also subjected to screening against four strains of bacteria using the agar dilution method. The extraction yields were up to 5.0% w/w and up to 8.6% w/w for supercritical fluid extraction with pure CO(2) and with ethyl acetate as co-solvent, respectively, while the low-pressure extraction indicates yields up to 24.0% w/w in the soxhlet extraction using water and aqueous mixture with 50% ethanol as solvents. The inhibitory activity of the extracts in gram-positive bacteria was significantly higher than in gram negative. The quantification and the identification of the extracts recovered were accomplished using GC/MS analysis. The most important components identified in the extract were artemetin, beta-sitosterol, alpha-humulene and beta caryophyllene, among others. PMID- 19683437 TI - Profiling of cellulose content in Indian seaweed species. AB - Cellulose contents were estimated in 12 seaweed samples belonging to different families e.g. red, brown and green, growing in Indian waters. Each cellulose sample was fractionated to yield alpha (alpha) and beta (beta) celluloses. Characterization was done using various analytical tools and results were validated by comparison with those of the cellulose obtained from Whatman filter paper No. 4. The greatest yields of cellulose (crude), alpha- and beta-cellulose were obtained from Gelidiella acerosa (13.65%), Chamaedoris auriculata (9.0%) and G. acerosa (3.10%). G. acerosa was also found to contain relatively high amount of alpha-cellulose (8.19%). The lowest cellulose contents were recorded from Kappaphycus alvarezii (2.00%) and Sarconema scinaioides (2.1%), while the latter contained the lowest alpha-, and beta-celluloses (1.0% and 0.30%, respectively). It appears that agarophytic and alginophytic algae contain high cellulose and alpha-cellulose contents, while the carrageenophyte contains low cellulose. The brown algae, in general contain high cellulose as well as alpha- and beta celluloses. PMID- 19683438 TI - Modeling the effects of pelleting on the logistics of distillers grains shipping. AB - The energy security needs of energy importing nations continue to escalate. It is clear that biofuels can help meet some of the increasing need for energy. Theoretically, these can be produced from a variety of biological materials, including agricultural residues (such as corn stover and wheat straw), perennial grasses, legumes, algae, and other biological materials. Currently, however, the most heavily utilized material is corn starch. Industrial fuel ethanol production in the US primarily uses corn, because it is readily converted into fuel at a relatively low cost compared to other biomass sources. The production of corn based ethanol in the US is dramatically increasing. As the industry continues to grow, the amount of byproducts and coproducts also increases. At the moment, the nonfermentable residues (which are dried and sold as distillers dried grains with solubles--DDGS) are utilized only as livestock feed. The sale of coproducts provides ethanol processors with a substantial revenue source and significantly increases the profitability of the production process. Even though these materials are used to feed animals in local markets, as the size and scope of the industry continues to grow, the need to ship large quantities of coproducts grows as well. This includes both domestic as well as international transportation. Value-added processing options offer the potential to increase the sustainability of each ethanol plant, and thus the industry overall. However, implementation of new technologies will be dependent upon how their costs interact with current processing costs and the logistics of coproduct deliveries. The objective of this study was to examine some of these issues by developing a computer model to determine potential cost ramifications of using various alternative technologies during ethanol processing. This paper focuses specifically on adding a densification unit operation (i.e., pelleting) to produce value-added DDGS at a fuel ethanol manufacturing plant. We have examined the economic implications of pelleting DDGS for varying DDGS production rates (100-1000 tons/d) and pelleting rates (0-100%), for a series of DDGS sales prices ($50-$200/ton). As the proportion of pelleting increases, the cost of transporting DDGS to distant markets drastically declines, because the rail cars can be filled to capacity. For example, at a DDGS sales price of $50/ton, 100% pelleting will reduce shipping costs (both direct and indirect) by 89% compared to shipping the DDGS in bulk form (i.e., no pelleting), whereas at a DDGS sales price of $200/ton, it will reduce costs by over 96%. It is clear that the sustainability of the ethanol industry can be improved by implementing pelleting technology for the coproducts, especially at those plants that ship their DDGS via rail. PMID- 19683439 TI - Effects of mixing system and pilot fuel quality on diesel-biogas dual fuel engine performance. AB - This paper describes results obtained from CI engine performance running on dual fuel mode at fixed engine speed and four loads, varying the mixing system and pilot fuel quality, associated with fuel composition and cetane number. The experiments were carried out on a power generation diesel engine at 1500 m above sea level, with simulated biogas (60% CH(4)-40% CO(2)) as primary fuel, and diesel and palm oil biodiesel as pilot fuels. Dual fuel engine performance using a naturally aspirated mixing system and diesel as pilot fuel was compared with engine performance attained with a supercharged mixing system and biodiesel as pilot fuel. For all loads evaluated, was possible to achieve full diesel substitution using biogas and biodiesel as power sources. Using the supercharged mixing system combined with biodiesel as pilot fuel, thermal efficiency and substitution of pilot fuel were increased, whereas methane and carbon monoxide emissions were reduced. PMID- 19683440 TI - Semi-continuous biohydrogen production as an approach to generate electricity. AB - In this work, a semi-continuous biological system was established to produce hydrogen and generate electricity by coupling the bioreactor to a fuel cell. Heat and acid pretreatments (at 35 and 55 degrees C) of a seed sludge used as inoculum were performed in order to increase hydrogen producers. Different initial glucose concentrations (IGC) were tested for heat pretreated inoculum at 35 degrees C to determine the optimum concentration of glucose that supported the highest hydrogen production. Results showed that the heat pretreated inoculums (35 degrees C) reached the highest hydrogen molar yield of 2.85 mol H(2)/mol glucose (0.014 L/h), which corresponds to the acetic acid pathway. At the optimum IGC (10 g/L, 35 degrees C) the hydrogen molar yield was 3.6 mol H(2)/mol glucose (0.023 L/h). The coupled bioreactor-fuel cell system yielded an output voltage of 1.06 V, power of 0.1 W (25 degrees C) and a current of 68 mA. The overall results suggest that high hydrogen molar yields can be obtained through the acetic acid pathway and that is feasible to generate electricity using hydrogen from the semi continuous bioreactor. PMID- 19683442 TI - Synthesis of novel pyrazole carboxamide derivatives and discovery of modulators for apoptosis or autophagy in A549 lung cancer cells. AB - A series of novel 3-aryl-1-arylmethyl-1H-pyrazole-5-carboxamide derivatives 3a-l, were synthesized by the reaction of 3-aryl-1-arylmethyl-1H-pyrazole-5-carbonyl chloride with substituted amine in excellent yields. The compounds 3e-h could suppress A549 lung cancer cell growth. More interestingly, compounds 3e and 3f might inhibit the A549 cell growth by inducing apoptosis; whereas compounds 3g and 3h with fluorine group might inhibit the A549 cell growth by inducing autophagy. PMID- 19683441 TI - Discovery of novel diarylketoxime derivatives as selective and orally active melanin-concentrating hormone 1 receptor antagonists. AB - Optimization of the lead 2a led to the identification of a novel diarylketoxime class of melanin-concentrating hormone 1 receptor (MCH-1R) antagonists. Our focus was directed toward improvement of hERG activity and metabolic stability. The representative derivative 4b showed potent and dose-dependent body weight reduction in diet-induced obese (DIO) C57BL/6J mice after oral administration. The synthesis and structure-activity relationships of the novel diarylketoxime MCH-1R antagonists are described. PMID- 19683443 TI - Discovery of isoxazole voltage gated sodium channel blockers for treatment of chronic pain. AB - A series of novel isoxazole voltage gated sodium channel blockers have been synthesized and evaluated. Substitutions on the benzylic position of benzamide were investigated to determine their effect on Na(v)1.7 inhibitory potency. The spirocyclobutyl substitution had the most significant enhancement on Na(v)1.7 inhibitory activity. PMID- 19683444 TI - 164th ENMC International workshop: 6th workshop on centronuclear (myotubular) myopathies, 16-18th January 2009, Naarden, The Netherlands. PMID- 19683445 TI - Hearing sensitivity in women following chemotherapy treatment for breast cancer: results from a pilot study. AB - PURPOSE: Our aim was to examine in a prospective pilot study whether standard adjuvant treatments for breast cancer can adversely affect hearing. METHODS: Eight pre/peri-menopausal women with breast cancer had middle ear analysis (tympanometry) and pure tone audiometry conducted prior to and 6 months following Fluorouracil, Epirubicin, Cyclophosphamide (FEC) or FEC plus taxotere chemotherapy treatments. RESULTS: The mean hearing thresholds in both ears showed an elevation (that is a decline) post chemotherapy treatment at 6 and 8kHz of between 20 and 30dB, which is graded as a mild hearing impairment at the higher frequency range. There were individual differences in pattern and grade within the group. CONCLUSIONS: The variability noted in the data is more than that would be anticipated for test-retest variance, suggesting that the hearing impairments are complex but genuine. The most likely cause of the reduction in hearing sensitivity is a change in oestrogen levels resulting from the breast cancer treatments. PMID- 19683446 TI - Neurosurgery for obsessive-compulsive disorder: contemporary approaches. AB - Surgery for psychiatric disorders has a controversial history. Traditionally, procedures were undertaken to physically disconnect or destroy certain areas of the brain thought to constitute critical components of the limbic pathways. The relatively recent advent of the much safer and non-destructive technique known as deep brain stimulation has coincided with a resurgence in interest in psychosurgery. Contemporary approaches to the surgical management of obsessive compulsive disorder are discussed, together with our current understanding of its pathophysiology. PMID- 19683447 TI - Polymorphism of HD and UCHL-1 genes in Huntington's disease. AB - This study analyzed the association between the polymorphism of the Huntington's disease (HD) and ubiquitin carboxyl-terminal hydrolase L1 (UCHL-1) genes and the age of HD onset. We examined the size of trinucleotide CAG repeats in the HD gene of 53 individuals from families with a history of HD, six unrelated HD patients, and 51 healthy controls. Polymerase chain reaction and restriction fragment length polymorphism was performed to examine UCHL-1 S18Y polymorphism prevalence in this group. We identified five HD patients in the families and four pre clinical HD patients in their high-risk offspring. The differences in S18Y allele prevalence between families and healthy controls were not statistically significant. The SY genotype was identified most frequently (prevalence >50%). The YY genotype was not identified in non-related HD patients, and the SS genotype had a higher prevalence than the SY genotype. The S allele was identified more frequently than the Y allele, and the difference with healthy controls was significant. Multiple linear regression analysis revealed that UCHL 1 S18Y polymorphism accounted for 15.6% of variance in the age of disease onset among 11 patients. The number of CAG repeats accounted for 71.4% of the variance. The size of CAG repeats in the HD gene is an important factor affecting the age at disease onset, but is not the only factor. UCHL-1 S18Y polymorphism is a modifier of HD with a modest regulatory role in the age at disease onset, suggesting that UCHL-1 may be involved in HD pathogenesis. PMID- 19683448 TI - The role of the standard 20 minute EEG recording in the comatose patient. AB - Non-convulsive seizures and non-convulsive status epilepticus (NCSE) are believed common in comatose patients and are suggested to worsen outcome. The purpose of this study was to prospectively evaluate outcome in patients in critical care units in whom NCSE was suspected to determine how often evidence of seizure activity existed based on an isolated standard 20 minute electroencephalogram (EEG) and to determine what clinical factors predicted outcome. We prospectively reviewed EEGs and clinical charts of patients admitted to a critical care unit at a tertiary care center who were suspected to have non-convulsive seizures. Outcomes were correlated with EEG findings, clinical factors, and acute therapies using univariate and multivariate logistic analyses. Of 189 patients, complete information was available in 169. Eighty-one (47.9%) patients died, 67 (39.6%) were discharged home, and 21 (12.4%) were discharged to long-term care. Four patients had electroencephalographic seizures, two of whom had no clinical manifestations (i.e. non-convulsive). On univariate analysis, increased age, an admitting diagnosis of cardiac arrest, a Glasgow Coma Scale (GCS) score8, and burst suppression were correlated significantly with poor outcome. A past history of seizures and unequivocal tonic-clonic convulsions were correlated significantly with a better outcome. On multivariate analysis, increased age, cardiac arrest, and a GCS score8 were associated with increased mortality (p<0.05). Clinical factors, including age, underlying etiology and GCS score are the most important predicators of outcome in coma. A standard 20 minute EEG did not correlate with a high detection rate of seizure activity. Furthermore, EEG patterns and treatment with anticonvulsant medications did not correlate with outcome. PMID- 19683449 TI - Synthesis and pharmacological evaluation of 6-naltrexamine analogs for alcohol cessation. AB - A series of substituted aryl amide derivatives of 6-naltrexamine, 3 designed to be metabolically stable were synthesized and used to characterize the structural requirements for their potency to binding and functional activity of human mu (mu), delta (delta) and kappa (kappa) opioid and nociceptin (NOP) receptors. Binding assays showed that 4-10 had subnanomolar K(i) values for mu and kappa opioid receptors. Functional assays for stimulation of [(35)S]GTPgammaS binding showed that several compounds acted as partial or inverse agonists and antagonists of the mu and delta, kappa opioid or NOP receptors. The compounds showed considerable stability in the presence of rat, mouse or human liver preparations and NADPH. The inhibitory activity on the functional activity of human cytochrome P450s was examined to determine any potential inhibition by 4-9. Only modest inhibition of CYP3A4, CYP2C9 and CYP2C19 was observed for a few of the analogs. As a representative example, radiolabeled 6 was examined in vivo and showed reasonable brain penetration. The inhibition of ethanol self administration in rats trained to self-administer a 10% (w/v) ethanol solution, utilizing operant techniques showed 5-8 to have very potent efficacy (ED(50) values 19-50 microg/kg). PMID- 19683450 TI - Design, synthesis and cruzain docking of 3-(4-substituted-aryl)-1,2,4-oxadiazole N-acylhydrazones as anti-Trypanosoma cruzi agents. AB - Research in recent years has demonstrated that the Trypanosoma cruzi cysteine protease cruzain (TCC) is a valid chemotherapeutic target, since inhibitors of this protease affect the pathology appropriately. By exploring the N acylhydrazones (NAH) as privileged structures usually present in antiparasitic agents, we investigated a library of 16 NAH bearing the 3-(4-substituted-aryl) 1,2,4-oxadiazole scaffold (NAH 3a-h, 4a-h). The in vitro bioactivity against epimastigote and trypomastigote forms of T. cruzi was evaluated, and some NAH under study exhibited antitrypanosomal activity at concentrations that are not toxic to mammalian cells. The series of compounds based on the 3-(4-substituted aryl)-1,2,4-oxadiazole scaffold revealed the remarkable importance of each substituent at the phenyl's 4-position for the inhibitory activity. Non-nitrated compounds 3a and 4e were found to be as potent as the reference drug, Benznidazole. In addition, the molecular origin of the antitrypanosomal properties for these series was investigated using docking studies of the TCC structure. PMID- 19683452 TI - Polymorphism of amyloid-beta fibrils and its effects on human erythrocyte catalase binding. AB - The Alzheimer's amyloid-beta (Abeta) peptide exists as a number of naturally occurring forms due to differential proteolytic processing of its precursor molecule. Many of the Abeta peptides of different lengths form fibrils in vitro, which often show polymorphisms in the fibril structure. This study presents a TEM based analysis of fibril formation by eighteen different Abeta peptides ranging in length from 5 to 43 amino acids. Spectrophotometric analysis of Congo red binding to the fibrillar material has been assessed and the binding of human erythrocyte catalase (HEC) to Abeta fibrils has also been investigated by TEM. The results show that a diverse range of Abeta peptides form fibrils and also bind Congo red. The ability of both Abeta 1-28 and Abeta 29-40 to form fibrils indicates that there are at least two fibril-forming domains within the full length Abeta 1-40 sequence, the ability of many Abeta peptides to form Congo red binding aggregates suggests that there may be up to 4 possible aggregation promoting domains. The binding of HEC was limited to Abeta forms containing residues 29-32. The differing capacities of fibrillar and ribbon-like structures may reflect the accessibility of the 29-32 region and suggest that HEC may be able to discriminate between different forms of Abeta fibrils. PMID- 19683451 TI - Omega-conotoxin GVIA mimetics based on an anthranilamide core: effect of variation in ammonium side chain lengths and incorporation of fluorine. AB - A number of omega-conotoxin GVIA mimetics based on an anthranilamide core were prepared and tested for their affinity for rat brain Ca(v)2.2 channels. Features such as the presence of hydroxyl and fluoro substituents on the tyrosine side chain mimic, the length of the chains on the lysine/arginine side chain mimics and the use of diguanidino and diamino substituents rather than mono guanidine/mono-amine substitution were examined. The diguanidinylated compounds proved to be the most active and deletion of the hydroxyl substituent had a limited influence on activity. The SAR associated with variation in the lysine/arginine side chain mimics was not strong. The introduction of a fluoro substituent into the tyrosine mimic produced the most active compound prepared in this study (2g), with an EC(50) at rat brain Ca(v)2.2 channels of 6 microM. PMID- 19683453 TI - Synthesis of carbon-11 labeled celecoxib derivatives as new candidate PET radioligands for imaging of inflammation. AB - Cyclooxygenase (prostaglandin endoperoxide synthase or COX) enzyme represents a particularly attractive target in inflammation processes for the development of both therapeutic agents and imaging agents. This study was designed to develop new radioligands for imaging of inflammation using the biomedical imaging technique positron emission tomography (PET). Carbon-11 labeled celecoxib derivatives, [(11)C]methyl 2-(4-(5-p-tolyl-3-(trifluoromethyl)-1H-pyrazol-1 yl)phenylsulfonamidooxy)acetate ([(11)C]6e), [(11)C]methyl 2-methyl-2-(4-(5-p tolyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)phenylsulfonamidooxy)propanoate ([(11)C]6f), [(11)C]methyl 2-(4-(5-(4-methoxyphenyl)-3-(trifluoromethyl)-1H pyrazol-1-yl)phenylsulfonamidooxy)acetate ([(11)C]6g), and [(11)C]methyl 2-methyl 2-(4-(5-(4-methoxyphenyl)-3-(trifluoromethyl)-1H-pyrazol-1 yl)phenylsulfonamidooxy)propanoate ([(11)C]6h), were prepared by O [(11)C]methylation of their corresponding precursors using [(11)C]CH(3)OTf under basic condition and isolated by a simplified solid-phase extraction (SPE) method in 50-60% radiochemical yields based on [(11)C]CO(2) and decay corrected to end of bombardment (EOB). The overall synthesis time from EOB was 15-20 min, the radiochemical purity was >99%, and the specific activity at end of synthesis (EOS) was 111-185 GBq/micromol. PMID- 19683455 TI - A fully echo-guided trans-apical aortic valve implantation. AB - The trans-apical aortic valve implantation (TA-AVI) is an established technique for high-risk patients requiring aortic valve replacement. Traditionally, preoperative (computed tomography (CT) scan, coronary angiogram) and intra operative imaging (fluoroscopy) for stent-valve positioning and implantation require contrast medium injections. To preserve the renal function in elderly patients suffering from chronic renal insufficiency, a fully echo-guided trans catheter valve implantation seems to be a reasonable alternative. We report the first successful TA-AVI procedure performed solely under trans-oesophageal echocardiogram control, in the absence of contrast medium injections. PMID- 19683454 TI - Spectral interference corrections for the measurement of (238)U in materials rich in thorium by a high resolution gamma-ray spectrometry. AB - In this study, the spectral interferences are investigated for the analytical peaks at 63.3 keV of (234)Th and 1001.0 keV of (234m)Pa, which are often used in the measurement of (238)U activity by the gamma-ray spectrometry. The correction methods are suggested to estimate the net peak areas of the gamma-rays overlapping the analytical peaks, due to the contribution of (232)Th that may not be negligible in materials rich in natural thorium. The activity results for the certified reference materials (CRMs) containing U and Th were measured with a well type Ge detector. The self-absorption and true coincidence-summing (TCS) effects were also taken into account in the measurements. It is found that ignoring the contributions of the interference gamma-rays of (232)Th and (235)U to the mixed peak at 63.3 keV of (234)Th ((238)U) leads to the remarkably large systematic influence of 0.8-122% in the measured (238)U activity, but in case of ignoring the contribution of (232)Th via the interference gamma-ray at 1000.7 keV of (228)Ac to the mixed peak at 1001 keV of (234m)Pa ((238)U) results in relatively smaller systematic influence of 0.05-3%, depending on thorium contents in the samples. The present results showed that the necessary correction for the spectral interferences besides self-absorption and TCS effects is also very important to obtain more accurate (238)U activity results. Additionally, if one ignores the contribution of (232)Th to both (238)U and (40)K activities in materials, the maximum systematic influence on the effective radiation dose is estimated to be ~6% and ~1% via the analytical peaks at 63.3 and 1001 keV for measurement of the (238)U activity, respectively. PMID- 19683456 TI - A huge aortic arch aneurysm mimicking massive pulmonary artery embolism. PMID- 19683457 TI - Open-heart surgery in premature and low-birth-weight infants--a single-centre experience. AB - OBJECTIVE: Because of their poor clinical status, infants may require surgery for congenital heart disease regardless of weight or prematurity. This retrospective review describes a single-centre experience with open-heart surgery in low-weight infants. METHODS: From November 1997 to December 2006, 411 open-heart surgery procedures were performed in neonates. This included 46 consecutive infants weighing less than 2500 g, who underwent cardiopulmonary bypass for correction of congenital heart defects (n=34) or Norwood stage I palliation of hypoplastic left heart syndrome (HLHS) (n=12). In the low-weight group were 23 males and 23 females with a median age of 10 days and a median weight of 2.26 kg (range: 1.28 2.49 kg). RESULTS: Early mortality was 8.2% in patients weighing more than 2.5 kg and 13% in the low-weight group. Within the low-weight group, weight at surgery, history of prematurity and prevalence of additional extracardiac malformations did not influence early mortality. At a median follow-up time of 32 months overall mortality was 21%. Thirty-four patients had a neurological follow-up examination 30 months postoperatively. Of the 34 survivors, 11 showed neurological deficits. CONCLUSIONS: In our patient population, early mortality was higher for infants weighing less than 2.5 kg. However, within the low-weight group, lower weight at surgery or history of prematurity was not associated with a higher mortality or bad neurological outcome. PMID- 19683459 TI - Lumbar and abdominal muscle activity during walking in subjects with chronic low back pain: support of the "guarding" hypothesis? AB - It has been hypothesized that changes in trunk muscle activity in chronic low back pain (CLBP) reflect an underlying "guarding" mechanism, which will manifest itself as increased superficial abdominal - and lumbar muscle activity. During a functional task like walking, it may be further provoked at higher walking velocities. The purpose of this cross sectional study was to investigate whether subjects with CLBP show increased co-activation of superficial abdominal - and lumbar muscles during walking on a treadmill, when compared to asymptomatic controls. Sixty-three subjects with CLBP and 33 asymptomatic controls walked on a treadmill at different velocities. Surface electromyography data of the erector spinae, rectus abdominis and obliquus abdominis externus muscles were obtained and averaged per stride. Results show that, compared to asymptomatic controls, subjects with CLBP have increased muscle activity of the erector spinae and rectus abdominis, but not of the obliquus abdominis externus. These differences in trunk muscle activity between groups do not increase with higher walking velocities. In conclusion, the observed increased trunk muscle activity in subjects with CLBP during walking supports the guarding hypothesis. PMID- 19683458 TI - Long-term cardiopulmonary exercise capacity after modified Fontan operation. AB - OBJECTIVE: Early circuit separation enhances the long-term success of Fontan haemodynamics. To test this hypothesis, we analysed the postoperative cardiopulmonary capacity in children and adults. PATIENTS: Spiroergometry was performed at least twice in 43 patients with a median age of 14 (range: 7-43) years, with a median time interval of 4.6 (1.1-10.4) years between early and late testing. Twenty-eight patients had been operated on in childhood and 15 as adults. The exercise capacity (W(max)) and oxygen consumption capacity (VO(2max)) were compared between children and adults. RESULTS: The VO(2max) in children early postoperatively was better than in adults (median 27.9 vs 22.9, p=0.032). Both VO(2max) (median 30.1 ml min(-1) kg(-1) vs 16.9 ml min(-1)kg(-1), p<0.001), and W(max) (median 2.2 W kg(-1) vs 1.4 W kg(-1), p<0.001) were significantly better in children late after surgery. In the patient group as a whole, there was a significant decrease of VO(2max) between early and later testing (median 26.5 l min(-1) kg(-1) vs 20.7 l min(-1) kg(-1), p<0.001). CONCLUSIONS: Fontan palliation in early childhood results in better cardiopulmonary capacity during long-term follow-up. Regular surveillance of the physical capacity by spiroergometry is indispensable for the supervision of patients with Fontan haemodynamics. PMID- 19683460 TI - Me or not me--an optimal integration of agency cues? AB - Recent work has demonstrated that the sense of agency is not only determined by efference-copy-based internal predictions and internal comparator mechanisms, but by a large variety of different internal and external cues. The study by Moore and colleagues [Moore, J. W., Wegner, D. M., & Haggard, P. (2009). Modulating the sense of agency with external cues. Conscious and Cognition] aimed to provide further evidence for this view by demonstrating that external agency cues might outweigh or even substitute efferent signals to install a basic registration of self-agency. Although the study contains some critical points that, so we argue, are central to a proper interpretation of the data, it hints at a new perspective on agency: optimal cue integration seems to be the key to a robust sense of agency. We here argue that this framework could allow integrating the findings of Moore and colleagues and other recent agency studies into a comprehensive picture of the sense of agency and its pathological disruptions. PMID- 19683461 TI - Anosognosia in Alzheimer's disease--the petrified self. AB - This paper reviews the literature concerning the neural correlates of the self, the relationship between self and memory and the profile of memory impairments in Alzheimer's disease (AD) and explores the relationship between the preservation of the self and anosognosia in this condition. It concludes that a potential explanation for anosognosia in AD is a lack of updating of personal information due to the memory impairments characteristic of this disease. We put forward the hypothesis that anosognosia is due in part to the "petrified self." PMID- 19683462 TI - Outcome of surgical treatment in non-lesional intractable childhood epilepsy. AB - PURPOSE: The aim of this study is to investigate seizure outcomes following epilepsy surgery in non-lesional neocortical pediatric epilepsy. METHODS: We performed a retrospective study on 27 patients with intractable childhood epilepsy who received epilepsy surgery between 1999 and 2006 at Sang-gye Paik Hospital and Severance Children's Hospital. None of the patients had any detectable lesions on MRI. Surgical outcome was assessed at least 2 years postoperatively; clinical characteristics, surgical outcome, pathologic findings, types of surgery, localizing features on SPECT, FDG-PET, and long-term video-EEG were reviewed. RESULTS: Eighteen patients (67%) demonstrated an Engel class I outcome postoperatively. The mean follow-up duration was 4.3 years (range, 2.2-9 years). Eighteen out of 27 cases (67%) showed focal localizing features on ictal SPECT, and 21 of 27 cases (78%) showed abnormal findings on PET. Single lobectomy was the most common procedure, and was performed in 20 patients (74%). Multilobar resection was performed in seven patients (26%). Cortical dysplasia was the most common finding during pathological examination, and was observed in 15 (56%) cases. In addition, gliosis was found in two patients (7%) and non-specific pathological findings were described in 10 patients (37%). CONCLUSIONS: In children with intractable epilepsy and a MRI demonstrating no abnormal lesions, epileptic surgery should be strongly considered when cortical pathology can be identified from other studies. PMID- 19683464 TI - Evidence for a novel vasospastic transmitter system, neuromedin U, in the equine digital circulation. AB - The brain-gut peptide neuromedin U (NMU) is a ligand for the G-protein-coupled receptors, NMU1 and NMU2. In humans, an extended form of this peptide, NMU-25, and the structurally related peptide, neuromedin S (NMS), both produce potent vasoconstriction in isolated blood vessels. The aim of this study was to determine whether NMU fulfilled criteria for controlling vasoreactivity in the equine digital circulation. NMU receptors were characterised in the equine digital artery and vein based on the pharmacological criteria of specific, saturable and high affinity binding. Immunoreactive peptide was detected in the equine digital artery and vein using anti-NMS antisera. [(125)I]-NMU-25 binding sites were localised to the smooth muscle layer and NMU-25 potently constricted the digital vein. This provides evidence for NMU as a transmitter in the equine digital circulation. PMID- 19683463 TI - Reversible neurological deficit after foam sclerotherapy. AB - A 37-year-old male was treated with 9 ml of 3% polidocanol foam, and he immediately reported photopsiae lasting a few minutes, though without migraine. Two hours after sclerotherapy, the patient developed speech disturbance for a few minutes. A pathological examination revealed nothing except a patent foramen ovale (PFO). Given the contrast between the high prevalence of PFO in general population and the extremely low incidence of neurological deficits after foam sclerotherapy, these deficits may only arise due to as-yet-unknown aetiology. PMID- 19683465 TI - A comparison of anatomical pain sites from a tertiary care sample: evidence of disconnect between functional and perceived disability specific to lower back pain. AB - Heterogeneity has been identified within chronic musculoskeletal pain (CMP) patient samples; however, investigations have typically focused on psychological constructs or coping (e.g., pain-related anxiety, catastrophizing) in this regard. Furthermore, studies to date have included either samples presenting with a specific anatomical site (e.g., only lower back pain) or a mix of anatomical sites (e.g., lower back, shoulder, or leg pain) as the primary pain complaint, without making comparisons based on the anatomical site of reported pain. For example, patients with chronic lower back pain (CLBP) may differ from those with chronic upper or lower extremity pain (ULEP) in presentation, recovery trajectory, and psychological variables. The current investigation explored whether systematic differences existed between patients participating in a multidisciplinary reconditioning third-party-payer program who have CLBP relative to patients with ULEP. Patients included those with CLBP (n=23; 35% women) or ULEP (e.g., arm, shoulder, leg, knee; n=28; 29% women). The ULEP group began and finished the program with more pain-related anxiety, more catastrophic thoughts, and more fearful cognitions than the CLBP group. There were no significant correlations between functional deficit and perceived levels of disability or associations between group and return to work status; however, there was an unexpected significant interaction between group and perceived disability. Specifically, CLBP patients reported increasing perceived disability despite improvements in functional deficit, whereas ULEP patients did not. These findings suggest a disconnect between perceived disability and function that may be specific to lower back pain. Implications and directions for future research are discussed. PMID- 19683466 TI - Growth and characterization of a new nonlinear optical crystal: [(18C6)Li][Cd(SCN)3]. AB - Single crystals of a new nonlinear optical compound [(18C6)Li][Cd(SCN)(3)][(18 crown-6-ether) lithium(I) trithiocyanate cadmium(II), CLTC], with dimension of 22 mm x 8 mm x 2 mm, were grown from aqueous solutions for the first time via evaporation technique. Solubility of CLTC has been determined for various temperatures. The grown crystals were characterized by single crystal X-ray diffraction (XRD) study, powder X-ray diffraction, FT-IR techniques, UV-vis, elemental analysis, EDS analyses and SHG test. Single crystal XRD study has been carried out to identify the lattice parameters and CLTC crystallizes in orthorhombic system. FT-IR studies confirm the functional groups present in the grown crystal. The mechanical properties of the grown crystals have been studied using Vickers microhardness tester. The optical second harmonic generation effect has been measured by using the Kurtz powder technique to be 2 times as large as that of KDP (KH(2)PO(4)) crystal. From the optical spectrum, CLTC has good optical transmission in the entire visible region, which is an essential requirement for a nonlinear crystal. PMID- 19683467 TI - Chikungunya virus of Asian and Central/East African genotypes in Malaysia. AB - BACKGROUND: Chikungunya virus (CHIKV) of the Central/East African genotype has caused large outbreaks worldwide in recent years. In Malaysia, limited CHIKV outbreaks of the endemic Asian and imported Central/East African genotypes were reported in 1998 and 2006. Since April 2008, an unprecedented nationwide outbreak has affected Malaysia. OBJECTIVE: To study the molecular epidemiology of the current Malaysian CHIKV outbreak, and to evaluate cross-neutralisation activity of serum from infected patients against isolates of Asian and Central/East African genotypes. STUDY DESIGN: Serum samples were collected from 83 patients presenting in 2008, and tested with PCR for the E1 gene, virus isolation, and for IgM. Phylogenetic analysis was performed on partial E1 gene sequences of 837bp length. Convalescent serum from the current outbreak and Bagan Panchor outbreak (Asian genotype, 2006) were tested for cross-neutralising activity against representative strains from each outbreak. RESULTS: CHIKV was confirmed in 34 patients (41.0%). The current outbreak strain has the A226V mutation in the E1 structural protein, and grouped with Central/East African isolates from recent global outbreaks. Serum cross-neutralisation activity against both Central/East African and Asian genotypes was observed at titres from 40 to 1280. CONCLUSIONS: The CHIKV strain causing the largest Malaysian outbreak is of the Central/East African genotype. The presence of the A226V mutation, which enhances transmissibility of CHIKV by Aedes albopictus, may explain the extensive spread especially in rural areas. Serum cross-neutralisation of different genotypes may aid potential vaccines and limit the effect of future outbreaks. PMID- 19683469 TI - Feasibility of the simultaneous ocular and cervical vestibular-evoked myogenic potentials in unilateral vestibular hypofunction. AB - OBJECTIVE: This study compared the results of combined and individual ocular vestibular-evoked myogenic potential (oVEMP) and cervical VEMP (cVEMP) tests in healthy subjects and patients with unilateral vestibular hypofunction to confirm the effectiveness of the combined oVEMP and cVEMP test. METHODS: Twenty healthy volunteers and 12 patients with unilateral vestibular hypofunction underwent individual oVEMP and cVEMP tests in a random order, and combined oVEMP and cVEMP test on another day. RESULTS: Twenty healthy subjects had 100% response rates for oVEMPs and cVEMPs in both individual and combined stimulation modes. Significant positive correlations exist between individual and combined oVEMPs/cVEMPs in terms of latencies and amplitudes. In 12 patients with unilateral vestibular hypofunction, differences in abnormal percentages of oVEMPs or cVEMPs were non significant. Additionally, the characteristic parameters of oVEMP/cVEMP among ears of healthy subjects, good and pathological ears of patients with unilateral vestibular hypofunction did not differ significantly, regardless of whether the individual or combined mode was used. CONCLUSIONS: The combined oVEMP and cVEMP test generates similar information to that obtained by individual mode, regardless of whether subjects are healthy or have unilateral vestibular hypofunction. SIGNIFICANCE: Simultaneous oVEMP and cVEMP tests may be a convenient screening tool for assessing crossed vestibulo-ocular reflex and ipsilateral sacculo-collic reflex, which definitely shortens the diagnostic test time. Thus, it may favor the large diffusion of these techniques. PMID- 19683468 TI - Auditory event-related potential changes in chronic occupational exposure to organophosphate pesticides. AB - OBJECTIVE: To determine whether chronic occupational exposure to organophosphates (OP) pesticides leads to cognitive impairment using event-related potentials (ERPs). METHODS: ERPs of 38 vegetable farmers applying OP pesticides and 35 controls were recorded using an auditory oddball paradigm. The N1, P2, N2 and P300 ERP components and the number of counting errors were compared between the groups. RESULTS: The farmers made significantly more counting errors than controls in the oddball task. The mixed model ANOVA of component latencies revealed a significant componentxgroup interaction, suggesting farmers had a greater delay in later ERP components. Intergroup comparisons of individual components showed significant delays in N2 and P300 latencies. Subsequent ANCOVA showed significant P300 delay even after adjusting for the latency of the preceding component, N2. Intergroup differences of P300 amplitudes were not significant, although there was limited evidence of a difference in scalp topography. CONCLUSION: Our findings indicate that chronic low-level occupational exposure to OP pesticides is associated with progressively increasing delay in successive ERP components, particularly P300. SIGNIFICANCE: Chronic exposure to OP pesticides may delay the neurophysiological processes underlying early stages of selective attention and late stages of sensory information processing that include stimulus evaluation and updating of working memory. PMID- 19683470 TI - Image-free assessment of protein translocation in live cells. AB - Protein translocation is a universal event shared by most cell signalling pathways to transmit signals between cell compartments. In recent years, the use of new fluorescence microscopy technologies combined with fluorescent probes- most often fluorescent proteins--and image analysis software has allowed the visualization and extensive analysis of such dynamic events in the context of the living cell. This review article focuses on emerging fluorescence approaches that tackle live cell protein translocation in the image-free context. Such methods are based on either protein-protein interactions or analysis of spatial diffusion of proteins by fluorescence intensity measurements. The potential benefits of intensity measurement on global cell populations versus image analysis of heterogeneous cell sample are discussed in the context of drug discovery applications. PMID- 19683471 TI - Cellular mechanisms of insulin resistance: role of stress-regulated serine kinases and insulin receptor substrates (IRS) serine phosphorylation. AB - Insulin receptor substrates (IRS) serine phosphorylation is a time-controlled physiological feedback mechanism in insulin signaling that is hijacked by metabolic and inflammatory stresses to promote insulin resistance. Kinases, including IKKbeta, JNK, ERK, mTOR, and S6K, activated by the inducers of insulin resistance induce uncontrolled IRS serine phosphorylation. Studies with genetically modified mice reveal that these kinases integrate signals from metabolic and inflammatory stresses in adipose tissue, liver, and hypothalamus leading to peripheral and central insulin resistance. Moreover, IKKbeta/NF-kappaB and JNK1 pathways in myeloid cells represent a core mechanism involved in inflammation linked to obesity. These kinases are thus potential drug targets against insulin resistance and the targeting of the IKKbeta/NF-kappaB or the JNK pathway may evolve into future diabetes medication. PMID- 19683472 TI - Identification of proteins from tuberculin purified protein derivative (PPD) by LC-MS/MS. AB - The tuberculin purified protein derivative (PPD) is a widely used diagnostic antigen for tuberculosis, however it is poorly defined. Most mycobacterial proteins are extensively denatured by the procedure employed in its preparation, which explains previous difficulties in identifying constituents from PPD to characterize their behaviour in B- and T-cell reactions. We here described a proteomics-based characterization of PPD from several different sources by LC MS/MS, which combines the solute separation power of HPLC, with the detection power of a mass spectrometer. The technique is able to identify proteins from complex mixtures of peptide fragments. A total of 171 different proteins were identified among the four PPD samples (two bovine PPD and two avium PPD) from Brazil and UK. The majority of the proteins were cytoplasmic (77.9%) and involved in intermediary metabolism and respiration (24.25%) but there was a preponderance of proteins involved in lipid metabolism. We identified a group of 21 proteins that are present in both bovine PPD but were not detected in avium PPD preparation. In addition, four proteins found in bovine PPD are absent in Mycobacterium bovis BCG vaccine strain. This study provides a better understanding of the tuberculin PPD components leading to the identification of additional antigens useful as reagents for specific diagnosis of tuberculosis. PMID- 19683473 TI - High granulocyte/lymphocyte ratio and paucity of NKT cells defines TB disease in a TB-endemic setting. AB - Most people infected with Mycobacterium tuberculosis, the causative agent of tuberculosis (TB) actually maintain a strong immune response and are able to control bacterial growth (deemed latently infected (LTBI)), while approximately 10% progress to disease resulting in almost 2 million deaths per year. Determining the immune 'footprint' at specific stages of infection and disease will allow for better diagnostics, treatments and ultimately development of new vaccine candidates. In this study we performed multi-factorial flow cytometry on fresh blood from 56 TB cases, 46 Tuberculin Skin Test (TST) positive (LTBI) and 39 TST negative household contacts. We found a highly significant increase in granulocytes and decrease in B cells and invariant (Valpha24+Vbeta11+) NKT cells in TB cases compared to TST+ contacts (p<0.0001, p=0.007 and p=0.01 respectively) which were restored to LTBI levels following 6 months of TB treatment. Using support vector analysis, we found a combination of granulocyte and lymphocyte and/or NKT cell proportions allowed almost 90% correct classification into M. tuberculosis infection or disease. This work has important public health benefits in regards to diagnosis and treatment of TB in sub-Saharan Africa and in furthering our understanding of the requirements for protective immunity to TB. PMID- 19683474 TI - TBrowse: an integrative genomics map of Mycobacterium tuberculosis. AB - Tuberculosis is one of the major infectious diseases causing morbidity and mortality in the developing world. Genome-wide experiments on Mycobacterium tuberculosis particularly H37Rv and many other strains has revealed a wealth of information on the pathogen. This has been complemented by computational methods for the analysis of genomic sequence. This genome-level information is scattered in individual databases and supplementary material of publications and is not easily amenable to integrative analysis and visualization. TBrowse is an attempt to create a starting resource for integrative analysis of the M. tuberculosis genome. This comprehensive database contains more than half a million data-points of genomic data systematically culled from online resources and publications and is organized into hundred tracks. The resource is built based on the Generic Model Organism Database Genome Browser, thus making it readily interoperable with other genome browser installations. TBrowse is enabled with tools for programmatic data access and interoperability with other similar resources through Distributed Annotation System. In addition the resource is interfaced with sequence analysis servers maintained by the National Center for Biotechnology Information and the University of California Santa Cruz. The resource is available at http://tbrowse.osdd.net. PMID- 19683475 TI - Quality assurance of the donor questionnaire and donor interview: a three year experience with an electronic donor questionnaire. AB - Quality assurance of the donor questionnaire and the donor interview must ensure that all relevant questions about donor eligibility are answered and documented in a reliable format. The use of the self explanatory, electronic donor information tool [EDIT], not only provided a harmonized and standardised system of quality assurance but also saved time and can be easily modified as guidelines change. This brief report highlights the principles of this tool and some of its potential benefits, as experienced over the 3 years since its introduction in Norway. PMID- 19683476 TI - Presurgical neuropsychological testing predicts cognitive and seizure outcomes after anterior temporal lobectomy. AB - We sought to determine significant predictors of seizure and cognitive outcome following surgery for epilepsy. Participants included 41 patients who had undergone anterior temporal lobectomy (ATL). Higher presurgical verbal/language scores and lower nonverbal memory scores were predictive of seizure-free status following ATL. Overall, the presurgical predictors were 93% accurate in discriminating between seizure-free and non-seizure-free patients postsurgery. Surgery in the nondominant-for-language hemisphere was predictive of higher postsurgical verbal/language and verbal memory scores. Higher presurgical visual/construction, nonverbal memory, and verbal/language scores were predictive of better postsurgical verbal/language functioning. Better presurgical verbal/language functioning was predictive of the same skills postsurgically as well as visual/construction outcomes. Exploratory analyses in a subset of participants (n=25) revealed that dominant and nondominant intracarotid amobarbital (Wada) memory scores added unique variance only for predicting nonverbal memory following ATL. Presurgical neuropsychological testing provides significant and unique information regarding postsurgical seizure freedom and cognitive outcome in patients who have undergone ATL. PMID- 19683477 TI - The development of the chromaffin cell lineage from the neural crest. AB - Chromaffin cells are neuroendocrine cells, which are highly specialized for the synthesis and release of multiple hormones. Like sympathetic neurons, which are essential, inter alia, for neural control of vascular tone, they are derivatives of the neural crest, a transient structure at the dorsal surface of the embryonic neural tube. Chromaffin cells and sympathetic neurons have many features in common, but are also distinct in several respects. This review provides a summary of similarities and differences regarding the development of chromaffin cells and sympathetic neurons, viewed from molecular and morphological perspectives. Two major, still not finally settled issues, are whether (1) the two related cell types arise from one common or two separate cell lineages of delaminating neural crest cells, (2) in the former case when does lineage segregation occur, and what are the molecules underlying their phenotypic diversification. PMID- 19683478 TI - DNA lesions sequestered in micronuclei induce a local defective-damage response. AB - Micronuclei are good markers of chromosome instability and, among other disturbances, are closely related to double-strand break induction. The ability of DNA lesions sequestered in the micronuclear bodies to activate the complex damage-signalling network is highly controversial since some repair factors have not been consistently detected inside micronuclei. In order to better understand the efficiency of the response induced by micronuclear DNA damage, we have analyzed the presence of DNA damage-response factors and DNA degradation markers in these structures. Radiation-induced DNA double-strand breaks produce a modification of chromatin structural proteins, such as the H2AX histone, which is rapidly phosphorylated around the break site. Strikingly, we have been able to distinguish two different phosphoH2AX (gammaH2AX) labelling patterns in micronuclei: discrete foci, indicating DSB presence, and uniform labelling affecting the whole micronucleus, pointing to genomic DNA fragmentation. At early post-irradiation times we observed a high fraction of micronuclei displaying gammaH2AX foci. Co-localization experiments showed that only a small fraction of the DSBs in micronuclei were able to properly recruit the p53 binding protein 1 (53BP1) and the meiotic recombination 11 (MRE11). We suggest that trafficking defects through the micronuclear envelope compromise the recruitment of DNA damage-response factors. In contrast to micronuclei displaying gammaH2AX foci, we observed that micronuclei showing a gammaH2AX extensive-uniform labelling were more frequently observed at substantial post-irradiation times. By means of TUNEL assay, we proved that DNA degradation was carried out inside these micronuclei. Given this scenario, we propose that micronuclei carrying a non-repaired DSB are conduced to their elimination, thus favouring chromosome instability in terms of allele loss. PMID- 19683479 TI - Pulmonary nocardiosis in cystic fibrosis. AB - BACKGROUND: The treatment of Nocardia species found in the sputum of cystic fibrosis patients is of unknown value. METHODS: We conducted a retrospective analysis of the impact of directed oral antibiotic therapy against Nocardia spp. isolated from the sputum of 17 cystic fibrosis patients over a 10-year period. Pulmonary Function Tests were used as the clinical indicator of the disease state and the data were analyzed by general linear mixed model statistics with univariate analysis. RESULTS: Pulmonary Function Test values of all patients studied showed no significant difference before, during, or after the antibiotic treatment period. Treatment groups did not differ from non-treatment groups. This held true for Forced Expiratory Volume over 1 s and Functional Vital Capacity analysis. In addition, individual patient analysis did not reveal any trends or outliers. CONCLUSIONS: Oral antibiotic therapy of cystic fibrosis patients colonized with Nocardia does not appear to affect clinical outcome. This suggests that deferring therapy may be an acceptable alternative and justifies conducting a future placebo controlled trial. In addition, this study model may be useful in analyzing the effect of therapy on other rare and difficult organisms, such as fungi and mycobacteria in the cystic fibrosis population. PMID- 19683480 TI - A label-free nano-liquid chromatography-mass spectrometry approach for quantitative serum peptidomics in Crohn's disease patients. AB - The identification of serum biomarkers for the diagnosis of inflammatory bowel diseases able to reduce the need for invasive tests represents a major goal in their therapy and follow-up. We report here a methodological approach for the evaluation of specific changes in the serum peptides abundance in healthy (H) and Crohn's disease (CD) subjects, based on a label-free LC ESI/Q-TOF differential mass spectrometry (MS) approach combined with targeted MS/MS analysis. The low molecular weight serum proteins were separated by RP nano-LC ESI/Q-TOF MS and the resulting datasets were aligned with msInspect software. The differently abundant peptides, evaluated using Proteios Software Environment, were identified by MS/MS analysis and database search. The identification of clusters of peptides resulting from proteins (such as fibrinogen-alpha) commonly involved in physiological processes lead to the evaluation of a possible role in CD of specific serum exoproteases. An assay based on synthetic peptides spiked into H, CD and ulcerative colitis (UC) serum samples as substrate, followed by MALDI MS and chemometric analysis of the metabolite patterns has been developed achieving a 100% discrimination between CD, UC and H subjects. The results are promising for the application of this approach as a simple tool for diagnostic aims and biomarker discovery in CD. PMID- 19683481 TI - Liposome-based immunoaffinity chromatographic assay for the quantitation of immunoglobulin E in human serum. AB - Immunoglobulin E (IgE)-mediated type I allergies affect over 25% of the world's population; they are among the most common diseases in developed countries. Therefore, simple and rapid in vivo and in vitro methods for diagnosing allergies are becoming increasingly important. In this paper, we demonstrate the feasibility of using sulforhodamine B, a fluorescent dye, entrapped inside immunoliposomes, the outer surfaces of which were sensitized with IgE, as a signal amplifier for the development of a simple, rapid, and inexpensive colorimetric affinity chromatographic immunoassay for the detection of total IgE in serum. This assay operates based on competition between standards (or human serum samples) containing IgE and IgE-sensitized immunoliposomes for the limited number of antigen binding sites of immobilized anti-IgE antibodies at the antigen capture (AC) zone on the nitrocellulose membranes. The color density of the AC zone is indirectly proportional to the number of IgE units present in the test sample. The detection limit of this liposome-based immunoaffinity chromatographic assay was 0.37ng in IgE-free serum solution (equivalent to 20microL of a 18.5ngmL(-1) solution). A commercially available ELISA kit was used as a reference method to validate the proposed assay through the analysis of three human serum samples. PMID- 19683482 TI - Rotational femoral osteotomies using an endomedullary saw. AB - INTRODUCTION: The goal of this retrospective study is to analyze a series of ten patients (11 osteotomies) who underwent closed femoral rotational osteotomy using an endomedullary saw; stabilization was achieved by a locked centromedullary nail. We report the indications, technical aspects, clinical and radiological results as well as intercurrent complications with this surgical technique. HYPOTHESIS: Femoral endomedullary osteotomy is a safe procedure to correct malrotations. PATIENTS AND METHODS: Eleven femoral rotational osteotomies using an endomedullary saw were performed on ten patients, between January 1999 and July 2007. The indications were post-traumatic rotational malunions or congenital rotational deformities. The angular deformity averaged 33.5 degrees (24 degrees 52 degrees ). They were divided into internal rotation (ten cases) or external rotation (one case). One patient required a bilateral rotational osteotomy because of a congenital femoral malrotation combined to bilateral trochlear dysplasia. Rotational correction was, in two patients, simultaneously associated with a closed lengthening osteotomy. Clinical and radiological follow-up averaged 4 years and 9 months (26-104 months). The angular corrections obtained by these rotation osteotomies were calculated by CAT scans. RESULTS: Ten out of eleven osteotomies allowed a correction within a 4 degrees range in relation to the physiological femoral neck anteversion values (or to the contralateral side in the case of a healthy opposite lower extremity). There was no bone, joint, skin, or soft tissues infection, no pseudoarthrosis and no delayed outgrowth. We observed a transient neurological complication in the area of the pudendal nerve, during a combined rotational-lengthening osteotomy, as well as a bilateral femoral fracture during the bilateral rotational osteotomy. In all patients, consolidation occurred within a 3- to 5-month delay. The subjective results showed that eight out of nine patients (one was lost to follow-up) were satisfied or very satisfied with their operation, their functional recovery and the aesthetic aspects of their scars. DISCUSSION: Closed rotational osteotomies in adults represent a reliable, effective, safe and reproducible procedure for the correction of femoral torsion problems, whether they are post-traumatic or congenital. These results can be obtained only by respecting the indications and by applying a rigorously planned technique, including expertise in centromedullary nailing. LEVEL OF EVIDENCE: Level IV, retrospective therapeutic study. PMID- 19683483 TI - G-308A TNF-alpha polymorphism is associated with an increased risk of hepatocellular carcinoma in the Turkish population: case-control study. AB - BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha) is a pro-inflammatory cytokine that may act as an endogenous tumor promoter. A genetic polymorphism of TNF-alpha gene at position -308 promoter region is involved in the regulation of expression level and has been found to be associated with susceptibility to various types of cancer. METHODS: To determine the association of the TNF-alpha gene G-308A polymorphism on the risk of hepatocellular carcinoma (HCC) in a Turkish population, a hospital-based case-control study was designed consisting of 110 diagnosis subjects with hepatocellular carcinoma and 110 cancer-free control subjects matched on age, gender, smoking and alcohol status. The genotype frequency of this polymorphism was determined by using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. RESULTS: The distribution G-308A genotype was significantly associated with the risk of HCC (p<0.001, odds ratio [OR]=4.75, 95% confidence interval [CI]=2.25-9.82 for -308 AA/GA genotypes versus GG genotype). CONCLUSION: We suggested that the presence of the high producer allele -308A in the TNF-alpha gene appears to be associated with an increased risk for the development of HCC in Turkish population. PMID- 19683484 TI - Obesity is associated with atypia in breast ductal lavage of women with proliferative breast disease. AB - BACKGROUND: Benign proliferative breast disease without atypia slightly increases breast cancer risk but there are currently few clinical options for breast cancer prevention in this group of women. METHODS: We conducted a pilot study of women with a past diagnosis of proliferative breast disease with a goal to determine if the characteristics of cells obtained by breast ductal lavage were related to nutritional factors. RESULTS: There were 57 women who enrolled. A total of 39 women yielded nipple aspirate fluid (NAF) samples and 36 underwent breast ductal lavage. Five of the lavage samples were acellular and 28 had at least 200 cells. Surprisingly, atypia was present in 11 women. Presence of atypia was associated with slight changes in morphometric features of the epithelial cells such as measures of circularity as obtained by image analysis, but the only variable significantly different in women with atypia (versus no atypia) was a higher mean body mass index. Body mass index was also significantly correlated with C reactive protein (CRP) levels in the nipple aspirate fluid, indicating that obesity might have a pro-inflammatory effect on the breast that can contribute to increased rates of atypia. CONCLUSIONS: Although the clinical significance of atypia in breast ductal lavage is uncertain, these results support further work on prevention of obesity as a strategy for reducing breast cancer risk. PMID- 19683485 TI - Absence of PIWIL2 (HILI) expression in human bladder cancer cell lines and tissues. AB - BACKGROUND: PIWIL2, a member of Argonaute family of proteins, is exclusively expressed in testis and functions in development and maintenance of germline stem cells. Recently, ectopic expression of PIWIL2 has been reported in a variety of human and mouse tumors. To investigate a potential involvement of PIWIL2 in human bladder cancer, we examined its expression in several human bladder cancer cell lines, normal uroepithelial cell cultures, and some bladder tissues. METHODS: Relative expression of PIWIL2 was determined by real-time quantitative RT-PCR in fifteen bladder carcinoma cell lines, six normal uroepithelial cell cultures and seventy tissue specimens of tumor, tumor margins and morphologically normal tissues of bladder. Specific primers for PIWIL2, TBP and GAPDH (as two internal controls) were used for reverse transcription polymerase chain reaction technique. RESULTS: Real-time qRT-PCR demonstrated high PIWIL2 expression in testis tissue, but at least 240-fold lower expression in all examined cell lines. The highest expression outside testis was observed in one of six primary cultures of normal uroepithelial cells, but even lower expression of PIWIL2 was detected in any of the examined tumor and non-tumor tissues. CONCLUSION: Lack of PIWIL2 expression in most tissues along with its aberrant expression in some tumors candidate the gene as an attractive tumor marker for some neoplasms. However, our study indicates that PIWIL2 does not play a role in carcinogenesis of human bladder carcinoma. PMID- 19683486 TI - General practitioner and specialist views on colonoscopic screening of first degree relatives of colorectal cancer patients. AB - INTRODUCTION: Barriers to targeted colonoscopic screening of first-degree relatives of colorectal cancer patients have been the subject of considerable literature, always as seen from the patient's viewpoint. In the specific context of screening individuals with a family history, physicians may also play a predominant role in motivating their colorectal cancer patients to pass on screening information to their relatives. The aim of this study was to examine the views of general practitioners and specialists regarding barriers and facilitators affecting participation of relatives in colonoscopic screening. METHODS: A qualitative study was conducted to collect and analyse information from two focus groups of 4 general practitioners and 9 gastroenterologists respectively, and from semi-structured individual interviews with 10 general practitioners, 2 surgeons and 3 oncologists. An extended focus group of 36 gastroenterologists was organized to validate the results. RESULTS: The main barriers to colonoscopic screening of FDRs were associated with lack of direct communication between physicians and FDRs. Physicians needed support in the task of informing index patients and persuading them to transfer information on screening to their FDRs. The general practitioners spoke of their expectations in terms of training in and motivation for promoting screening and the gastroenterologists expressed the wish for patient education material specific to colonoscopy and for systematic post-colonoscopy consultations to inform patients about their results, follow-up and screening their relatives. CONCLUSION: The findings, notably the need for specific education materials and for training to improve the motivation of physicians, will help to develop effective interventions intended to increase participation in screening. PMID- 19683488 TI - Can the protein occupancy landscape show the topologically isolated chromosomal domains in the E. coli genome?: An exciting prospect. AB - In a recent issue of Molecular Cell, Vora et al. (2009) introduce an in vivo protein occupancy display (IPOD) technology and propose that the domains extensively occupied by proteins with inactive transcription in bacteria are mainly topologically isolated chromosomal domains. PMID- 19683487 TI - Examining the association between cigarette smoking and colorectal cancer using historical case-control data. AB - BACKGROUND: The majority of recent, well-designed studies have shown that long term cigarette smoking increases colorectal cancer risk, but older studies with shorter durations of exposure often found no association. This study aimed to examine colorectal cancer risk by smoking exposure using data collected in the late-1950s and early-1960s. METHODS: This case-control study examined colorectal cancer risk by lifetime smoking history. There were 1365 patients who visited Roswell Park Cancer Institute (RPCI) between 1957 and 1965 diagnosed with primary, incident colorectal cancers that were matched to 4096 malignancy-free controls on gender and age. Odds ratios were calculated using separate logistic regression models for each smoking exposure, while controlling for other tobacco use, county of residence, race, age, gender, and body mass index (BMI). RESULTS: The adjusted OR for individuals who reported their greatest level of smoking to be more than 1 pack/day was 0.87 (95% CI=0.67-1.15). Among those who smoked 42 or more years, the adjusted OR was 0.89 (95% CI=0.68-1.15) compared to those who never smoked. For individuals who smoked more than 45 pack-years, the OR was 0.92 (95% CI=0.72-1.19). The results did not differ significantly by gender, although men had considerably greater exposure compared to women. Results also did not differ by colorectal sub-site. CONCLUSION: No association was found between long term cigarette smoking and colorectal cancer risk. These results are in accord with studies that followed cohorts throughout the 1950s and 1960s. Methodological limitations, such as missing data on covariates and the higher incidence of smoking-related illness in a hospital setting, may have contributed to the null results found in this study. Prolonged population exposure to cigarettes and perhaps a changing product may explain why more recent studies have reported a positive association between smoking and colorectal cancer. PMID- 19683489 TI - The SAGA continues...to the end. AB - In this issue of Molecular Cell, Atanassov et al. (2009) show that the GCN5 containing SAGA complex regulates telomere function via deubiquitination and stabilization of the telomere repeat binding factor TRF1. PMID- 19683490 TI - SOSS1/2: Sensors of single-stranded DNA at a break. AB - In this issue of Molecular Cell, Huang et al. (2009) describe two heterotrimeric single-stranded DNA binding complexes, SOSS1 and SOSS2, that function downstream of the MRN complex to promote DNA repair and the G2/M checkpoint. PMID- 19683491 TI - PCI complexes: Beyond the proteasome, CSN, and eIF3 Troika. AB - The bipartite PCI domain serves as the principal scaffold for proteasome lid, CSN, and eIF3, complexes that influence protein life span. PCI domains are also found in newly identified complexes directing nucleic acid regulation. The breadth of functions associated with the extended PCI family is a factor of shared subunits, among them a common factor Sem1/DSS1 that facilitates complex assembly. PMID- 19683492 TI - Reconstitution of the death-inducing signaling complex reveals a substrate switch that determines CD95-mediated death or survival. AB - The death-inducing signaling complex (DISC) is critical for initiation of death receptor-mediated apoptosis; however, paradoxically, CD95 also signals for cell survival. Here, we reconstitute a functional DISC using only purified CD95, FADD, and procaspase-8 and unveil a two-step activation mechanism involving both dimerization and proteolytic cleavage of procaspase-8 that is obligatory for death-receptor-induced apoptosis. Initially, dimerization yields active procaspase-8 with a very restricted substrate repertoire, limited to itself or c FLIP. Proteolytic cleavage is then required to fully activate caspase-8, thereby permitting DISC-mediated cleavage of the critical exogenous apoptotic substrates, caspase-3 and Bid. This switch in catalytic activity and substrate range is a key determinant of DISC signaling, as cellular expression of noncleavable procaspase 8 mutants, which undergo DISC-mediated oligomerization, but not cleavage, fails to initiate CD95-induced apoptosis. Thus, using the reconstituted DISC, we have delineated a crucial two-step activation mechanism whereby activated death receptor complexes can trigger death or survival. PMID- 19683493 TI - Structure of the s5a:k48-linked diubiquitin complex and its interactions with rpn13. AB - Degradation by the proteasome typically requires substrate ubiquitination. Two ubiquitin receptors exist in the proteasome, S5a/Rpn10 and Rpn13. Whereas Rpn13 has only one ubiquitin-binding surface, S5a binds ubiquitin with two independent ubiquitin-interacting motifs (UIMs). Here, we use nuclear magnetic resonance (NMR) and analytical ultracentrifugation to define at atomic level resolution how S5a binds K48-linked diubiquitin, in which K48 of one ubiquitin subunit (the "proximal" one) is covalently bonded to G76 of the other (the "distal" subunit). We demonstrate that S5a's UIMs bind the two subunits simultaneously with a preference for UIM2 binding to the proximal subunit while UIM1 binds to the distal one. In addition, NMR experiments reveal that Rpn13 and S5a bind K48 linked diubiquitin simultaneously with subunit specificity, and a model structure of S5a and Rpn13 bound to K48-linked polyubiquitin is provided. Altogether, our data demonstrate that S5a is highly adaptive and cooperative toward binding ubiquitin chains. PMID- 19683494 TI - Negative regulation of the EGFR-MAPK cascade by actin-MAL-mediated Mig6/Errfi-1 induction. AB - We analyzed the G-actin-regulated transcriptome by gene expression analysis using previously characterized actin-binding drugs. We found many known MAL/MRTF dependent target genes of serum response factor (SRF), as well as additional directly regulated genes. Surprisingly, several putative antiproliferative target genes were identified, including mig6/errfi-1, a negative regulator of the EGFR family. Mig6 induction occurred through actin-MAL-SRF signaling, and MAL was inducibly recruited to and activated a mig6 promoter element. Upregulation of Mig6 by lipid agonists such as LPA and S1P or actin drugs involved MAL and correlated with decreased activation of EGFR, MAPK/Erk, and c-fos. Mig6 depletion restored EGFR signaling and provided a proliferative advantage. Overexpression of MAL exhibited strong antiproliferative effects requiring the domains for SRF binding and transactivation, which supports antagonistic functions of MAL on growth-promoting signals. Our results show the existence of negatively acting transcriptional networks between pro- and antiproliferative signaling pathways toward SRF. PMID- 19683495 TI - Ribosomal protein S7 is both a regulator and a substrate of MDM2. AB - MDM2 associates with ribosomal protein S7, and this interaction is required to inhibit MDM2's E3 ligase activity, leading to stabilization of MDM2 and p53. Notably, the MDM2 homolog MDMX facilitates the inhibition of MDM2 E3 ligase activity by S7. Further, ablation of S7 inhibits MDM2 and p53 accumulation induced by different stress signals in some cell types. Thus, ribosomal/nucleolar stress is likely a key integrating event in DNA damage signaling to p53. Interestingly, S7 is itself a substrate for MDM2 E3 ligase activity both in vitro and in vivo. An S7-ubiquitin fusion protein (S7-Ub) selectively inhibits MDM2 degradation of p53 and is unaffected by MDMX. S7-Ub promotes apoptosis to a greater extent than S7 alone. This indicates that MDM2 ubiquitination of S7 is involved in sustaining the p53 response. Thus, S7 functions as both effector and affector of MDM2 to ensure a proper cellular response to different stress signals. PMID- 19683496 TI - Cdk1 participates in BRCA1-dependent S phase checkpoint control in response to DNA damage. AB - Cdk2 and cdk1 are individually dispensable for cell-cycle progression in cancer cell lines because they are able to compensate for one another. However, shRNA mediated depletion of cdk1 alone or small molecule cdk1 inhibition abrogated S phase cell-cycle arrest and the phosphorylation of a subset of ATR/ATM targets after DNA damage. Loss of DNA damage-induced checkpoint control was caused by a reduction in formation of BRCA1-containing foci. Mutation of BRCA1 at S1497 and S1189/S1191 resulted in loss of cdk1-mediated phosphorylation and also compromised formation of BRCA1-containing foci. Abrogation of checkpoint control after cdk1 depletion or inhibition in non-small-cell lung cancer cells sensitized them to DNA-damaging agents. Conversely, reduced cdk1 activity caused more potent G2/M arrest in nontransformed cells and antagonized the response to subsequent DNA damage. Cdk1 inhibition may therefore selectively sensitize BRCA1-proficient cancer cells to DNA-damaging treatments by disrupting BRCA1 function. PMID- 19683497 TI - Two-color cell array screen reveals interdependent roles for histone chaperones and a chromatin boundary regulator in histone gene repression. AB - We describe a fluorescent reporter system that exploits the functional genomic tools available in budding yeast to systematically assess consequences of genetic perturbations on gene expression. We used our Reporter-Synthetic Genetic Array (R SGA) method to screen for regulators of core histone gene expression. We discovered that the histone chaperone Rtt106 functions in a pathway with two other chaperones, Asf1 and the HIR complex, to create a repressive chromatin structure at core histone promoters. We found that activation of histone (HTA1) gene expression involves both relief of Rtt106-mediated repression by the activity of the histone acetyltransferase Rtt109 and restriction of Rtt106 to the promoter region by the bromodomain-containing protein Yta7. We propose that the maintenance of Asf1/HIR/Rtt106-mediated repressive chromatin domains is the primary mechanism of cell-cycle regulation of histone promoters. Our data suggest that this pathway may represent a chromatin regulatory mechanism that is broadly used across the genome. PMID- 19683498 TI - Gcn5 and SAGA regulate shelterin protein turnover and telomere maintenance. AB - Histone acetyltransferases (HATs) play important roles in gene regulation and DNA repair by influencing the accessibility of chromatin to transcription factors and repair proteins. Here, we show that deletion of Gcn5 leads to telomere dysfunction in mouse and human cells. Biochemical studies reveal that depletion of Gcn5 or ubiquitin-specific protease 22 (Usp22), which is another bona fide component of the Gcn5-containing SAGA complex, increases ubiquitination and turnover of TRF1, a primary component of the telomeric shelterin complex. Inhibition of the proteasome or overexpression of USP22 opposes this effect. The USP22 deubiquitinating module requires association with SAGA complexes for activity, and we find that depletion of Gcn5 compromises this association in mammalian cells. Thus, our results indicate that Gcn5 regulates TRF1 levels through effects on Usp22 activity and SAGA integrity. PMID- 19683499 TI - yFACT induces global accessibility of nucleosomal DNA without H2A-H2B displacement. AB - FACT has been proposed to function by displacing H2A-H2B dimers from nucleosomes to form hexasomes. Results described here with yeast FACT (yFACT) suggest instead that nucleosomes are reorganized to a form with the original composition but a looser, more dynamic structure. First, yFACT enhances hydroxyl radical accessibility and endonuclease digestion in vitro at sites throughout the nucleosome, not just in regions contacted by H2A-H2B. Accessibility increases dramatically, but the DNA remains partially protected. Second, increased nuclease sensitivity can occur without displacement of dimers from the nucleosome. Third, yFACT is required for eviction of nucleosomes from the GAL1-10 promoter during transcriptional activation in vivo, but the preferential reduction in dimer occupancy expected for hexasome formation is not observed. We propose that yFACT promotes a reversible transition between two nucleosomal forms, and that this activity contributes to the establishment and maintenance of the chromatin barrier as well as to overcoming it. PMID- 19683500 TI - Histone chaperone spt16 promotes redeposition of the original h3-h4 histones evicted by elongating RNA polymerase. AB - Nucleosomes are surprisingly dynamic structures in vivo, showing transcription independent exchange of histones H2A-H2B genome-wide and exchange of H3-H4 mainly within the promoters of transcribed genes. In addition, nucleosomes are disrupted in front of and reassembled behind the elongating RNA polymerase. Here we show that inactivation of histone chaperone Spt16 in yeast results in rapid loss of H2B and H3 from transcribed genes but also from inactive genes. In all cases, histone loss is blocked by a transcription inhibitor, indicating a transcription dependent event. Thus, nucleosomes are efficiently evicted by the polymerase but do not reform in the absence of Spt16. Yet exchange of nucleosomal H2B with free histones occurs normally, and, unexpectedly, incorporation of new H3 increases at all loci tested. This points to Spt16 restoring normal nucleosome structure by redepositing the displaced H3-H4 histones, thereby preventing incorporation of new histones and perhaps changes in histone modification patterns associated with ongoing transcription. PMID- 19683502 TI - Public and private sector treatment of malaria in Lao PDR. AB - This study aimed to examine the care-seeking choices for treatment of a febrile illness compatible with malaria in the public and private sectors in Lao PDR. We conducted interviews with 745 heads of household in 14 villages in the Sekong province, using a structured-questionnaire. We asked each about who the care providers were for febrile illness episodes affecting their household members during the past year. If patients used more than one care-provider for a single episode over a period of time, we identified patterns of the care-sequences for the initial and subsequent care choices. Then, we analyzed the relationship between the initial care choices and secondary care choices for care-providers by Chi-square test, categorizing care-providers into public (hospital, health centre, and village health volunteer) and private care-providers (private pharmacy, informal retailer, faith healing and herbs). As a result, we found that 624 patients sought care at least once, 255 (40.9%) twice, and 66 (10.6%) three times or more during a single episode. Of 138 patients who started with a public care-provider and then sought a secondary care, 71 (51.4%) switched to a private care-provider. In contrast, of 117 patients who started with a private care provider and then sought a secondary care, 82 (70.1%) switched to a public care provider (p<0.001). In conclusion, although most patients who failed being treated by a private care-provider switched to a public one, some exclusively relied on care within the private sector. An intervention is necessary to make the private sector an integral component of malaria treatment in Lao PDR. PMID- 19683503 TI - Genes of cathepsin L-like proteases in Trypanosoma rangeli isolates: markers for diagnosis, genotyping and phylogenetic relationships. AB - We have sequenced genes encoding cathepsin L-like (CatL-like) cysteine proteases from isolates of Trypanosoma rangeli from humans, wild mammals and Rhodnius species of Central and South America. Phylogenetic trees of sequences encoding mature CatL-like enzymes of T. rangeli and homologous genes from other trypanosomes, Leishmania spp. and bodonids positioned sequences of T. rangeli (rangelipain) closest to T. cruzi (cruzipain). Phylogenetic tree of kinetoplastids based on sequences of CatL-like was totally congruent with those derived from SSU rRNA and gGAPDH genes. Analysis of sequences from the CatL-like catalytic domains of 17 isolates representative of the overall phylogenetic diversity and geographical range of T. rangeli supported all the lineages (A-D) previously defined using ribosomal and spliced leader genes. Comparison of the proteolytic activities of T. rangeli isolates revealed heterogeneous banding profiles of cysteine proteases in gelatin gels, with differences even among isolates of the same lineage. CatL-like sequences proved to be excellent targets for diagnosis and genotyping of T. rangeli by PCR. Data from CatL-like encoding genes agreed with results from previous studies of kDNA markers, and ribosomal and spliced leader genes, thereby corroborating clonal evolution, independent transmission cycles and the divergence of T. rangeli lineages associated with sympatric species of Rhodnius. PMID- 19683501 TI - SOSS complexes participate in the maintenance of genomic stability. AB - Proteins that bind to single-stranded DNA (ssDNA) are essential for DNA replication, recombinational repair, and maintenance of genomic stability. Here, we describe the characterization of an ssDNA-binding heterotrimeric complex, SOSS (sensor of ssDNA) in human, which consists of human SSB homologs hSSB1/2 (SOSS B1/2) and INTS3 (SOSS-A) and a previously uncharacterized protein C9orf80 (SOSS C). We have shown that SOSS-A serves as a central adaptor required not only for SOSS complex assembly and stability, but also for facilitating the accumulation of SOSS complex to DNA ends. Moreover, SOSS-depleted cells display increased ionizing radiation sensitivity, defective G2/M checkpoint, and impaired homologous recombination repair. Thus, our study defines a pathway involving the sensing of ssDNA by SOSS complex and suggests that this SOSS complex is likely involved in the maintenance of genome stability. PMID- 19683504 TI - Metabolomic analysis of biofluids from rats treated with Aconitum alkaloids using nuclear magnetic resonance and gas chromatography/time-of-flight mass spectrometry. AB - The Aconitum alkaloids aconitine, mesaconitine, and hypaconitine are the main toxic components in a commonly used traditional Chinese herbal medicine Fu Zi. To provide guidelines for the safe use of this medicine, metabolic changes in Wistar rats caused by these compounds were investigated by means of integrated analysis of two metabonomic approaches: (1)H nuclear magnetic resonance (NMR) and gas chromatography/time-of-flight mass spectrometry (GC/TOF-MS). Rats were given a single dose of aconitine, mesaconitine, hypaconitine, or vehicle. The largest metabolic changes were observed 6 h after treatment. Every group receiving a dose had higher urine concentrations of glucose, acetate, dimethylglycine, succinate, and alanine and had lower concentrations of creatinine, citrate, 2-oxoglutarate, N-acetylated metabolites, and trimethylamine-N-oxide (TMAO) than did the control group. These results may reflect the perturbation of renal tubular function within the first 24 h after treatment. The results also revealed a larger perturbation of metabolic profiles in the aconitine group than in the mesaconitine and hypaconitine groups, illustrating how these alkaloids exhibit different toxicities. An analysis of plasma samples collected 7 days postdose showed that there were higher levels of lactate, alanine, and lipids along with lower levels of glucose, beta-hydroxybutyrate, and creatine in the plasma of the aconitine and mesaconitine groups than there were in the control and hypaconitine groups. The GC/TOF-MS data from the plasma samples showed that the number of metabolites, with significant changes or with a tendency to change, in the aconitine and mesaconitine groups were dissimilar, suggesting a possible difference in the acute toxicity mechanisms of these alkaloids. PMID- 19683505 TI - A reverse-phase liquid chromatography/mass spectrometry method for the analysis of high-molecular-weight fructooligosaccharides. AB - Many important crop and forage plants accumulate polymeric water-soluble carbohydrates as fructooligosaccharides (or fructans). We have developed an improved method for the analysis of the full fructan complement in plant extracts based on porous graphitized carbon chromatography coupled to negative electrospray ionization mass spectrometry. By the use of profile data collection and multiple charge state ions, the effective mass range of the ion trap was extended to allow for the analysis of very high-molecular-weight oligosaccharides. This method allows the separation and quantification of isomeric fructan oligomers ranging from degree of polymerization (DP) 3 to DP 49. PMID- 19683506 TI - Interactions across the interface contribute the stability of homodimeric 3alpha hydroxysteroid dehydrogenase/carbonyl reductase. AB - The dimerization of 3alpha-hydroxysteroid dehydrogenase/carbonyl reductase was studied by interrupting the salt bridge interactions between D249 and R167 in the dimeric interface. Substitution of alanine, lysine and serine for D249 decreased catalytic efficiency 30, 1400 and 1.4-fold, and lowered the melting temperature 6.9, 5.4 and 7.6 degrees C, respectively. The mutated enzymes have the dimeric species but the equilibrium between monomer and dimer for these mutants varies from each other, implying that these residues might contribute differently to the dimer stability. Thermal and urea-induced unfolding profiles for wild-type and mutant enzymes appeared as a two-state transition and three-state transition, respectively. In addition, mutation on D249 breaks the salt bridges and causes different effects on the loss of enzymatic activity for D249A, D249K and D249S mutants in the urea-induced unfolding profiles. Hence, D249 at the dimeric interface in 3alpha-HSD/CR is essential for conformational stability, oligomeric integrity and enzymatic activity. PMID- 19683507 TI - EPR spectroscopy and electrospray ionization mass spectrometry reveal distinctive features of the iron site in leukocyte 12-lipoxygenase. AB - The procedure for the expression and purification of recombinant porcine leukocyte 12-lipoxygenase using Escherichia coli [K.M. Richards, L.J. Marnett, Biochemistry 36 (1997) 6692-6699] was updated to make it possible to produce enough protein for physical measurements. Electrospray ionization tandem mass spectrometry confirmed the amino acid sequence. The redox properties of the cofactor iron site were examined by EPR spectroscopy at 25K following treatment with a variety of fatty acid hydroperoxides. Combination of the enzyme in a stoichiometric ratio with the hydroperoxides led to a g4.3 signal in EPR spectra instead of the g6 signal characteristic of similarly treated soybean lipoxygenase 1. Native 12-lipoxygenase was also subjected to electrospray ionization mass spectrometry. There was evidence for loss of the mass of an iron atom from the protein as the pH was lowered from 5 to 4. Native ions in these samples indicated that iron was lost without the protein completely unfolding. PMID- 19683509 TI - Concerted bifunctionality of the dCTP deaminase-dUTPase from Methanocaldococcus jannaschii: a structural and pre-steady state kinetic analysis. AB - Two mutant dCTP deaminase-dUTPases from Methanocaldococcus jannaschii were crystallised and the crystal structures were solved: E145A in complex with the substrate analogue alpha,beta-imido-dUTP and E145Q in complex with diphosphate. Both mutant enzymes were defect in the deaminase reaction and had reduced dUTPase activity. In the structure of E145Q in complex with diphosphate, the diphosphate occupied the same position as the beta- and gamma-phosphoryls of the nucleotide analogue in the E145A complex. The C-terminal region that is unresolved in the apo-form of the enzyme was ordered in both complexes and closed over the active site by interacting with the phosphate backbone of the nucleotide or with the diphosphate. A magnesium ion was readily observed to complex with all three phosphoryls in the nucleotide complex or with the diphosphate. A water molecule that is likely to be involved in the nucleotidyl diphosphorylase reaction was observed in the E145A:alpha,beta-imido-dUTP complex and positioned similarly as in the monofunctional trimeric dUTPase. A comparison of the active sites of the bifunctional enzyme and the monofunctional family members, dCTP deaminase and dUTPase, suggests similar reaction mechanisms. The similar side chain conformations in the deaminase site between the nucleotide and diphosphate complexes indicated a concerted re-arrangement, or induced fit, of the whole active site promoted by enzyme and nucleotide phosphoryl interactions. A pre steady state kinetic analysis of the bifunctional reaction and the dUTPase half reaction supported a conformational change upon substrate binding in both reactions and a concerted catalytic step for the bifunctional reaction. PMID- 19683508 TI - Characterization of a blue-copper protein, auracyanin, of the filamentous anoxygenic phototrophic bacterium Roseiflexus castenholzii. AB - A blue-copper protein auracyanin of the filamentous anoxygenic phototroph Roseiflexus castenholzii was purified and characterized. Genomic sequence analysis showed that R. castenholzii has only one auracyanin, whereas Chloroflexus aurantiacus is known to have two auracyanins, A and B. Absorption spectrum of the Roseiflexus auracyanin was similar to that of auracyanin B of C. aurantiacus. On the other hand, ESR spectrum of the Roseiflexus auracyanin resembles that of auracyanin A of C. aurantiacus. These results suggest that the blue-copper protein auracyanin from R. castenholzii shares features with each of auracyanin A and B. Amino acid sequence alignment of auracyanins from filamentous anoxygenic phototrophs also demonstrated the chimeral feature of the primary structure of the Roseiflexus auracyanin, i.e., auracyanin A-like amino-terminal characteristics and auracyanin B-like one-residue spacing at the Cu-binding loop in the carboxyl-terminus. PMID- 19683510 TI - Internalization of MUC1 by anti-MUC1 antibody from cell membrane through the macropinocytotic pathway. AB - MUC1 is a type I transmembrane glycoprotein aberrantly overexpressed in various cancer cells. It is thought to serve as a physical barrier from the extracellular environment and also as a receptor for various extracellular molecules. However, little is known about the fate of MUC1 during and after the interaction with these molecules. In the present study, we used anti-MUC1 antibody as an interacting molecule and investigated the cellular trafficking of MUC1. Our results showed that: (1) anti-MUC1 antibody was internalized only in MUC1 expressing cells and triggered internalization and down-regulation of MUC1; (2) the internalization of MUC1 by anti-MUC1 antibody required the cytoplasmic tail of MUC1 and was suppressed by inhibitors of Na(+)/H(+) exchanger, and caveola/raft-dependent internalization, but not by an inhibitor of clathrin dependent internalization. We conclude that antibody-induced internalization of MUC1 involves the macropinocytotic pathway. PMID- 19683511 TI - Defects in embryonic development of EGLN1/PHD2 knockdown transgenic mice are associated with induction of Igfbp in the placenta. AB - The HIF (hypoxia inducible factor) hydroxylases EGNL1/PHD2 has been implicated in embryonic development. Here we knocked down EGNL1 in vivo by injecting one-cell murine zygotes with lentivirus-containing RNAi. Progeny with demonstrated EGLN1 inhibition had elevated EPO production and erythropoiesis in vivo. The partial inhibition of EGLN1 in utero is embryonic lethal in some, but not all mice on gestation day 14, and is associated with defects in placental and heart development, similar to those noted in the EGLN1 knockout mouse. Importantly, the in utero inhibition of EGNL1 varied greatly between the embryo proper and the placenta. Using this as a tool we show that the embryopathic effects are associated with knockdown of EGNL1 and the associated induction of Igfbp1 (insulin-like growth factor binding protein-1) mRNA in the placenta, but not the embryo. PMID- 19683512 TI - Analysis of the role of the HIF hydroxylase family members in erythropoiesis. AB - HIF (hypoxia-inducible factor) hydroxylases have been implicated in EPO (erythropoietin) production and erythropoiesis. Here we examined the role of each of the three EGLN family members and the HIF asparaginyl hydroxylase FIH (factor inhibiting HIF) in EPO production. We examined the effect of inhibiting individual as well as combinations of HIF hydroxylases with RNAi. We found that inhibition of EGLN1 (egl nine homolog 1) as well as other members of the EGLN family (EGLN2 and EGLN3) led to accumulative EPO production in vitro. We then knocked down EGNL1 in vivo by injecting one-cell murine zygotes with lentivirus containing RNAi. Progeny with demonstrated EGLN1 inhibition had elevated EPO production and erythropoiesis in vivo. Among all the in vitro and in vivo studies, no or minimal VEGF (vascular endothelial growth factor) mRNA or protein stimulation resulted from inhibition of EGLN1. PMID- 19683513 TI - Inhibition of human P-glycoprotein transport and substrate binding using a galantamine dimer. AB - The human multidrug resistance transporter P-glycoprotein (P-gp) prevents the entry of compounds into the brain by an active efflux mechanism at the blood brain barrier (BBB). Treatment of neurodegenerative diseases, therefore, has become a challenge and the development of new reversible inhibitors of P-gp is pertinent to overcome this problem. We report the design and synthesis of a crosslinked agent based on the Alzheimer's disease treatment galantamine (Gal-2) that inhibits P-gp-mediated efflux from cultured cells. Gal-2 was found to inhibit the efflux of the fluorescent P-gp substrate rhodamine 123 in cancer cells that over-express P-gp with an IC(50) value of approximately 0.6 microM. In addition, Gal-2 was found to inhibit the efflux of therapeutic substrates of P gp, such as doxorubicin, daunomycin and verapamil with IC(50) values ranging from 0.3 to 1.6 microM. Through competition experiments, it was determined that Gal-2 modulates P-gp mediated efflux by competing for the substrate binding sites. These findings support a potential role of agents, such as Gal-2, as inhibitors of P-gp at the BBB to augment treatment of neurodegenerative diseases. PMID- 19683514 TI - Ferritin reporter used for gene expression imaging by magnetic resonance. AB - Magnetic resonance imaging (MRI) is a minimally invasive way to provide high spatial resolution tomograms. However, MRI has been considered to be useless for gene expression imaging compared to optical imaging. In this study, we used a ferritin reporter, binding with biogenic iron, to make it a powerful tool for gene expression imaging in MRI studies. GL261 mouse glioma cells were over expressed with dual-reporter ferritin-DsRed under beta-actin promoter, then gene expression was observed by optical imaging and MRI in a brain tumor model. GL261 cells expressing ferritin-DsRed fusion protein showed enhanced visualizing effect by reducing T2-weighted signal intensity for in vitro and in vivo MRI studies, as well as DsRed fluorescence for optical imaging. Furthermore, a higher contrast was achieved on T2-weighted images when permeating the plasma membrane of ferritin-DsRed-expressing GL261. Thus, a ferritin expression vector can be used as an MRI reporter to monitor in vivo gene expression. PMID- 19683515 TI - Lupeol triterpene, a novel diet-based microtubule targeting agent: disrupts survivin/cFLIP activation in prostate cancer cells. AB - Recently we showed Lupeol, a triterpene, found in fruits and vegetables inhibits the growth of tumors originated from human androgen-sensitive prostate cancer (CaP) cells and decreases the serum-PSA levels in a mouse model. Here, we provide evidence that Lupeol inhibits the growth of androgen-sensitive as well as androgen-insensitive CaP cells by inducing G2/M cell cycle arrest without exhibiting any toxicity to normal human prostate epithelial cells (PrEC) at the doses at which it kills cancer cells. We observed that Lupeol treatment to LNCaP and DU145 cells resulted in a dose-dependent (i) decrease in the protein levels of Cyclins-A, -B1, -D1, -D2, -E2, cyclin-dependent kinase (cdk)-2 and (ii) increase in the protein level of CDK-inhibitor p21. Since G2/M cell cycle phase is regulated by microtubule assembly, we investigated effect of Lupeol on microtubule assembly, its regulation and down-stream targets in CaP cells. Lupeol treatment significantly modulated the level of (i) microtubule components alpha tubulin and beta-tubulin, (ii) microtubule-regulatory protein stathmin, and (iii) microtubule-regulatory down-stream target/pro-survival protein survivin. Lupeol treatment also decreased the level of anti-apoptotic protein cFLIP. Finally, Lupeol was observed to significantly decrease the transcriptional activation of survivin and cFLIP genes in CaP cells. We conclude that the Lupeol-induced growth inhibition of CaP cells is a net outcome of simultaneous effects on stathmin, cFLIP, and survivin which results in the disruption of microtubule assembly. We suggest that Lupeol alone or as an adjuvant to other microtubule agents could be developed as a potential agent for the treatment of human CaP. PMID- 19683517 TI - Analytical assay characterization for 1-108 pro-B-type natriuretic peptide on the BioPlex 2200 analyzer. PMID- 19683516 TI - Generation of oxidant response to copper and iron nanoparticles and salts: Stimulation by ascorbate. AB - The present work describes a two-stage approach to analyzing combustion-generated samples for their potential to produce oxidant stress. This approach is illustrated with the two commonly encountered transition metals, copper and iron. First, their abilities to generate hydroxyl radical were measured in a cell-free, phosphate-buffered saline solution containing ascorbate and/or citrate. Second, their abilities to induce heme oxygenase-1 in cultured human epidermal keratinocytes were assessed in cell culture. Combustion-generated copper oxide nanoparticles were active in both assays and were found to be soluble in culture medium. Depletion of glutathione in the cells or loading the cells with ascorbate greatly increased heme oxygenase-1 induction in the presence of copper. By contrast, iron oxide nanoparticles were active in the phosphate-buffered saline but not in cell culture, and they aggregated in culture medium. Soluble salts of copper and iron exhibited the same contrast in activities as the respective combustion-generated particles. The results suggest that the capability of combustion-generated environmental samples to produce oxidant stress can be screened effectively in a two step process, first in phosphate-buffered saline with ascorbate and subsequently in epithelial cell culture for those exhibiting activity initially. The results also point to an unanticipated interaction in cells of oxidant stress-generating metals with an antioxidant (ascorbate) that is usually missing in culture medium formulations. Thus, ascorbate supplementation of cultured human cells is likely to improve their ability to model the in vivo effects of particulate matter containing copper and other redox-active metals. PMID- 19683518 TI - Glutamate dehydrogenase activity in lymphocytes of B-cell chronic lymphocytic leukaemia patients. AB - OBJECTIVES: To investigate the pattern of glutamate dehydrogenase (GLDH) activity, GLUD1 and GLUD2 expressions in peripheral blood mononuclear cells (PBMC) of untreated B-chronic lymphocytic leukemia (B-CLL) in healthy individuals (HI) and patients with infectious mononucleosis (IM). DESIGN AND METHODS: GLDH activity was determined in a supernatant obtained from pelleted PBMC. GLUD1 and GLUD2 mRNA expression was determined using a quantitative real-time polymerase chain reaction. CD19(+) B cells from PBMC were purified by using positive selection. RESULTS: The highest GLDH activity was found in PBMC of the B-CLL group followed by the HI group and IM group. The PBMC GLDH activity was higher in 60% of the B-CLL patients according to the established reference interval for our HI (2.17-5.70 microkat/g protein). The greater GLDH activity was also found in the CD19(+) cell preparation of the B-CLL patients (two of the three) but not in HI (n=3). The median value of GLUD1 expression was highest in the IM group (n=11), followed by the HI (n=14) and B-CLL groups (n=59) (median 4.69/3.78, P<0.005 and 4.69/2.91, P<0.0005, respectively). GLUD2 expression was not significantly different between groups. CONCLUSIONS: The increased GLDH activity is specific for the PBMC of B-CLL patients. The GLUD1 but not the GLUD2 gene expression pattern is different between the PBMC of IM and B-CLL patients. PMID- 19683519 TI - A national survey on pediatric critical values used in clinical laboratories across Canada. AB - OBJECTIVE: Notification of critical values to clinical staff is an important post analytical process in all acute care clinical laboratories. No data are available however on how laboratories obtain or establish critical values, particularly in pediatric settings. This study was designed to examine and compare critical values used for pediatric patients in biochemistry laboratories in Canada and assess potential interlaboratory variability. DESIGN AND METHODS: Fourteen clinical laboratories, including two in pediatric hospitals and twelve in hospitals caring for both children and adults, participated in a survey that included 14 pre-selected STAT chemistry tests and 19 pre-selected therapeutic drug monitoring (TDM) tests. RESULTS: Among fourteen chemistry tests, good agreement was observed for critical values used for sodium and pH at both low and high levels within 14 participant laboratories. Significant interlaboratory variability existed for glucose critical values at the high end, magnesium at high end, and PO2 at the low end. For 19 TDM tests, the majority of laboratories did not have alert values to report values over the therapeutic level but not toxic. For critical values greater than the toxic range, significant variability existed at both trough and peak levels among laboratories surveyed. When asked to provide the source for critical values established at each site, only a limited number of laboratories identified their sources as either internal decision or published references. CONCLUSION: Although all laboratories have established and routinely use critical values to alert clinical staff, considerable variability exists in both the critical limits reported as well as the source of such values. There is a clear need for new national efforts to standardize pediatric critical value reporting and establish evidence-based critical limits for all medical laboratories across Canada. PMID- 19683520 TI - Neural development in Onychophora (velvet worms) suggests a step-wise evolution of segmentation in the nervous system of Panarthropoda. AB - A fundamental question in biology is how animal segmentation arose during evolution. One particular challenge is to clarify whether segmental ganglia of the nervous system evolved once, twice, or several times within the Bilateria. As close relatives of arthropods, Onychophora play an important role in this debate since their nervous system displays a mixture of both segmental and non-segmental features. We present evidence that the onychophoran "ventral organs," previously interpreted as segmental anlagen of the nervous system, do not contribute to nerve cord formation and therefore cannot be regarded as vestiges of segmental ganglia. The early axonal pathways in the central nervous system arise by an anterior-to-posterior cascade of axonogenesis from neuronal cell bodies, which are distributed irregularly along each presumptive ventral cord. This pattern contrasts with the strictly segmental neuromeres present in arthropod embryos and makes the assumption of a secondary loss of segmentation in the nervous system during the evolution of the Onychophora less plausible. We discuss the implications of these findings for the evolution of neural segmentation in the Panarthropoda (Arthropoda+Onychophora+Tardigrada). Our data best support the hypothesis that the ancestral panarthropod had only a partially segmented nervous system, which evolved progressively into the segmental chain of ganglia seen in extant tardigrades and arthropods. PMID- 19683521 TI - Notch signal is sufficient to direct an endothelial conversion from non endothelial somitic cells conveyed to the aortic region by CXCR4. AB - During the early formation of the dorsal aorta, the first-forming embryonic vessel in amniotes, a subset of somitic cells selected as presumptive angioblasts, migrates toward the dorsal aorta, where they eventually differentiate into endothelial cells. We have recently shown that these processes are controlled by Notch signals (Sato, Y., Watanabe, T., Saito, D., Takahashi, T., Yoshida, S., Kohyama, J., Ohata, E., Okano, H., and Takahashi, Y., 2008. Notch mediates the segmental specification of angioblasts in somites and their directed migration toward the dorsal aorta in avian embryos. Dev. Cell 14, 890 901.). Here, we studied a possible link between Notch and chemokine signals, SDF1/CXCR4, the latter found to be dominantly expressed in developing aorta/somites. Although CXCR4 overexpression caused a directed migration of somitic cells to the aortic region in a manner similar to Notch, no positive epistatic relationships between Notch and SDF1/CXCR4 were detected. After reaching the aortic region, the CXCR4-electroporated cells exhibited no endothelial character. Importantly, however, once provided with Notch activity, they could successfully be incorporated into developing vessels as endothelial cells. These findings were obtained combining the tetracycline-inducible gene expression method with the transposon-mediated stable gene transfer technique. We conclude that Notch activation is sufficient to direct naive mesenchymal cells to differentiate into endothelial cells once the cells are conveyed to the aortic region. PMID- 19683522 TI - Rapid large artery remodeling following the administration and withdrawal of calcium channel blockers in spontaneously hypertensive rats. AB - Chronic treatment with several antihypertensive agents, including calcium channel blockers, may interfere with remodeling of large arteries and increased arterial stiffness. We hypothesize that even a short, seven-day administration of calcium channel blockers might alter an aortic remodeling in spontaneously hypertensive rat (SHR). Male SHR and normotensive WKY rats (n=14 each) were treated by either vehicle, vasculoselective calcium channel blocker nifedipine (1mg/kg/day) or cardiac/vascular calcium channel blockers diltiazem (5mg/kg/day) or verapamil (4 mg/kg/day, n=6 for each treatment) subcutaneously twice daily for seven days. Additional SHR rats were randomized for termination 24, 72 or 120 h (n=5 each) after the withdrawal of nifedipine. Systolic blood pressure was measured by tail cuff and thoracic aorta was collected for histomorphometric and functional analysis including acetylcholine-induced endothelium-dependent relaxation. Seven day administration of diltiazem and nifedipine, but not verapamil decreased blood pressure in SHR. All drugs significantly attenuated abnormal aortic wall thickness, cross-sectional area and media-to-lumen ratio, but only nifedipine improved impaired endothelium-dependent relaxation. Following the withdrawal of nifedipine, all measured parameters returned back to control SHR values within 72 h. Seven-day treatment with distinct calcium channel blockers attenuates hypertensive remodeling of aorta, which might be, in case of nifedipine, reactivated even by a very short withdrawal of the drug. Therefore, vasculoprotection by calcium channel blockers is not restricted to a prolonged blood pressure modulation, but occurs rapidly. These findings could be relevant for an intervention in augmented vascular stiffness and related cardiovascular risk. PMID- 19683524 TI - Characterization of the MAL2-positive compartment in oligodendrocytes. AB - Oligodendrocytes (OLs), the myelin-producing cells of the central nervous system, segregate different surface subdomains at the plasma membrane as do other differentiated cells such as polarized epithelia and neurons. To generate the complex membrane system that characterizes myelinating OLs, large amounts of membrane proteins and lipids need to be synthesized and correctly targeted. In polarized epithelia, a considerable fraction of apical proteins are transported by an indirect pathway involving a detour to the basolateral membrane before being internalized and transported across the cell to the apical membrane by a process known as transcytosis. The apical recycling endosome (ARE) or its equivalent, the subapical compartment (SAC), of hepatocytes is an intracellular trafficking station involved in the transcytotic pathway. MAL2, an essential component of the machinery for basolateral-to-apical transcytosis, is an ARE/SAC resident protein. Here, we show that, after differentiation, murine oligodendrocyte precursor and human oligodendroglioma derived cell lines, Oli-neu and HOG, respectively, up-regulate the expression of MAL2 and accumulate it in an intracellular compartment, exhibiting a peri-centrosomal localization. In these oligodendrocytic cell lines, this compartment shares some of the main features of the ARE/SAC, such as colocalization with Rab11a, sensitivity to disruption of the microtubule cytoskeleton with nocodazole, and lack of internalized transferrin. Therefore, we suggest that the MAL2-positive compartment in oligodendrocytic cells could be a structure analogous to the ARE/SAC and might have an important role in the sorting of proteins and lipids for myelin assembly during oligodendrocyte differentiation. PMID- 19683523 TI - Effects of rolipram and diazepam on the adaptive changes induced by morphine withdrawal in the hypothalamic paraventricular nucleus. AB - A role for the cyclic AMP systems in the development of morphine dependence has been previously reported. In this study we investigated whether morphine dependence was inhibited by phosphodiesterase (PDE) 4 inhibitors rolipram and diazepam. Dependence on morphine was induced by a 7-day s.c. implantation of morphine pellets. On day 8, morphine withdrawal was precipitated by an injection of naloxone. In order to determine the effect of rolipram and diazepam rats were injected with these drugs once daily for seven days as well as 30 min before of naloxone injection. When opioid withdrawal was precipitated, an enhanced noradrenaline turnover and increased level of cyclic AMP and cyclic GMP in the hypothalamic paraventricular nucleus (PVN) were observed 30 min after naloxone administration. Moreover, c-Fos expression was induced in the PVN after naloxone precipitated morphine withdrawal. Co-administration of rolipram or diazepam with morphine during the pre-treatment period, significantly reduced the signs of withdrawal, the enhancement of noradrenaline turnover and the increase in cyclic AMP. However, these inhibitors did not modify either levels of cyclic GMP or c Fos expression in the PVN. These findings demonstrate that co-administration of rolipram or diazepam with morphine attenuate the withdrawal syndrome and suggest that these compounds may prevent the up-regulation of the cyclic AMP pathway and the associated increase in cyclic AMP level in morphine-withdrawn rats. PMID- 19683526 TI - Silencing of TNF-alpha receptors coordinately suppresses TNF-alpha expression through NF-kappaB activation blockade in THP-1 macrophage. AB - Persistently elevated level of TNF-alpha has been implicated in several inflammatory disorders, however, its autocrine production through TNF-alpha receptors signaling is poorly understood. Here we report that simultaneous silencing of TNF-receptors, R1 and R2 by DNAzyme or siRNA suppressed TNF-alpha expression more efficiently than silencing them individually in lipopolysaccharides (LPS) stimulated THP-1 macrophages. Co-silencing of TNF receptors also inhibited TNF-alpha induced NF-kappaB activation to a higher extent. It was further observed that NF-kappaB inhibitor but not c-Jun N-terminal kinase inhibitor (SP600125) suppressed TNF-alpha expression. All these results suggest that TNF-alpha expression is regulated by synergistic signaling of TNF receptors through downstream NF-kappaB activation. PMID- 19683525 TI - alpha3beta1 integrin promotes cell survival via multiple interactions between 14 3-3 isoforms and proapoptotic proteins. AB - Laminin-5 and alpha3beta1 integrin promote keratinocyte survival; however, the downstream signaling pathways for laminin-5/alpha3beta1 integrin-mediated cell survival had not been fully established. We report the unexpected finding of multiple interactions between 14-3-3 isoforms and proapoptotic proteins in the survival signaling pathway. Ln5-P4 motif within human laminin-5 alpha3 chain promotes cell survival and anti-apoptosis by inactivating Bad and YAP. This effect is achieved through the formation of 14-3-3zeta/p-Bad and 14-3-3sigma/p YAP complexes, which is initiated by alpha3beta1 integrin and FAK/PI3K/Akt signaling. These complexes result in cytoplasmic sequestration of Bad and YAP and their subsequent inactivation. An increase in Akt1 activity in cells induces 14-3 3zeta and sigma, p-Bad, and p-YAP, promoting cell survival, whereas decreasing Akt activity suppresses the same proteins and inhibits cell survival. Suppression of 14-3-3zeta with RNA-interference inhibits cell viability and promotes apoptosis. These results reveal a new mechanism of cell survival whereby the formation of 14-3-3zeta/p-Bad and 14-3-3sigma/p-YAP complexes is initiated by laminin-5 stimulation via the alpha3beta1 integrin and FAK/PI3K/Akt signaling pathways, thereby resulting in cell survival and anti-apoptosis. PMID- 19683527 TI - Genetic and pharmacological evidence of intraneuronal Abeta accumulation in APP transgenic mice. AB - Intraneuronal punctate immunostaining in Alzheimer's disease brain and amyloid beta precursor protein (APP) transgenic mice has been suggested to represent Abeta, but this is somewhat controversial. Here we show that both biochemical Abeta levels and intraneuronal immunostaining are reduced in APP transgenic mice when gamma-secretase is inhibited. Moreover, BACE-1 deficient APP transgenic mice show neither Abeta production nor intraneuronal immunostaining. Our findings suggest that the punctate immunostaining with APP antibodies is due to Abeta that has accumulated inside neurons. Similar type of intraneuronal Abeta accumulation, which precedes senile plaque formation, may link Abeta to tauopathy and neurodegeneration in Alzheimer's disease pathogenesis. PMID- 19683528 TI - PANDER binds to the liver cell membrane and inhibits insulin signaling in HepG2 cells. AB - PANDER is a cytokine co-secreted with insulin from islet beta-cells. To date, the physiological function of PANDER remains largely unknown. Here we show that PANDER binds to the liver membrane by (125)I-PANDER saturation and competitive binding assays. In HepG2 cells, pre-treatment with PANDER ranging from 4 pM to 4 nM for 8h resulted in a maximal inhibition of insulin-stimulated activation of insulin receptor and insulin receptor substrate 1 by 52% and 63%, respectively. Moreover, PANDER treatment also reduced insulin-stimulated PI3K and pAkt levels by 55% and 48%, respectively. In summary, we have identified the liver as a novel target for PANDER, and PANDER may be involved in the progression of diabetes by regulating hepatic insulin signaling pathways. PMID- 19683529 TI - MCL-1ES, a novel variant of MCL-1, associates with MCL-1L and induces mitochondrial cell death. AB - Myeloid cell leukemia-1 (MCL-1L) is a pro-survival member of the BCL-2 family that promotes cell survival. In this study, we identify a new splicing variant of human MCL-1 that encodes MCL-1ES (extra short). Sequence analysis indicates that this variant results from splicing within the first coding exon of MCL-1 at a non canonical GC-AG donor-acceptor pair. The deduced sequence of MCL-1ES encodes a protein of 197 amino acids, and the PEST (proline, glutamic acid, serine, and threonine) motifs present in MCL-1L are absent. MCL-1ES interacts with MCL-1L and induces mitochondrial cell death, suggesting that alternative splicing of MCL-1 may control the fate of cells. PMID- 19683530 TI - Probing for primary functions of prohibitin in Trypanosoma brucei. AB - Prohibitins (PHBs) 1 and 2 are small conserved proteins implicated in a number of functions in the mitochondrion, as well as in the nucleus of eukaryotic cells. The current understanding of PHB functions comes from studies of model organisms such as yeast, worm and mouse, but considerable debate remains with regard to the primary functions of these ubiquitous proteins. We exploit the tractable reverse genetics of Trypanosoma brucei, the causative agent of African sleeping sickness, in order to specifically analyse the function of PHB in this highly divergent eukaryote. Using inducible RNA interference (RNAi) we show that PHB1 is essential in T. brucei, where it is confined to the cell's single mitochondrion forming a high molecular weight complex. PHB1 and PHB2 appear to be indispensible for mitochondrial translation. Their ablation leads to a decrease in mitochondrial membrane potential, however no effect on the level of reactive oxygen species was observed. Flagellates lacking either PHB1 or both PHB1 and PHB2 exhibit significant morphological changes of their organelle, most notably its inflation. Even long after the loss of the PHB proteins, mtDNA was unaltered and mitochondrial cristae remained present, albeit displaced to the periphery of the mitochondrion, which is in contrast to other eukaryotes. PMID- 19683531 TI - Age-related infection and transmission patterns of human cysticercosis. AB - Neurocysticercosis is recognised as an important but neglected cause of epilepsy in developing countries where the parasite occurs. Data on the transmission dynamics of the parasite in endemic areas are scarce. Individuals living in these areas are likely to be highly exposed to the parasite, but relatively few of them develop active infections. The present study aimed to describe and gain insights into changes in antibody responses and infection patterns related to age and/or gender in a south Ecuadorian rural population by combining antibody and antigen serological data with demographic characteristics. In 25% of the population, antibodies to Taenia solium cysticerci were detected whilst 2.9% had circulating parasite antigens. The proportion of antibody-positive individuals increased significantly until the age of 40years to become stable in older individuals. A rule-based simulation model was developed to explain these variations and to reflect the dynamics of exposure to, and transmission of, the parasite. In contrast, the proportion of people presenting circulating parasite antigens, reflecting an active infection, was significantly higher in people older than 60years. Immunosenescence could explain such an observation since a weaker immune system in the elderly would facilitate the establishment and maintenance of viable cysticerci compared with fully immunocompetent younger individuals. This work points out the role of the immune system in the development of cysticercosis within an exposed population and highlights new essential issues in understanding the transmission dynamics of the parasite, its incidence and the resulting immunological response at a population level. PMID- 19683532 TI - A monoclonal rat anti-mouse EMAP II antibody that functionally neutralizes pro- and mature-EMAP II in vitro. AB - EMAP II is an endothelial cell and monocyte activating proinflammatory cytokine, which has been demonstrated to induce endothelial cell apoptosis. In order to analyze its role in disease models linked to inflammation and endothelial cell death, we aimed to develop a neutralizing antibody against mouse EMAP II. Therefore, we generated rat monoclonal anti-mouse EMAP II antibodies by immunization with recombinant full length, mouse pro-EMAP II protein. We could identify by ELISA, hybridoma clones from fusion with mouse myeloma SP2/0 cells which produced antibodies recognizing both full length and mature EMAP II. We further characterized one antibody, M7/1 and demonstrated its ability to detect both EMAP II forms in Western blotting and to neutralize EMAP II directed migration of human peripheral blood monocytes as well as EMAP II induced apoptosis of tumor and endothelial cells. We conclude that this antibody can be useful to both target and analyze murine disease models, in which EMAP II may be involved. PMID- 19683534 TI - Delayed rectifier K(+) currents and cardiac repolarization. AB - The two components of the cardiac delayed rectifier current have been the subject of numerous studies since firstly described. This current controls the action potential duration and is highly regulated. After identification of the channel subunits underlying IKs, KCNQ1 associated with KCNE1, and IKr, HERG, their involvement in human cardiac channelopathies have provided various models allowing the description of the molecular mechanisms of the KCNQ1 and HERG channels trafficking, activity and regulation. More recently, studies have been focusing on the unveiling of different partners of the pore-forming proteins that contribute to their maturation, trafficking, activity and/or degradation, on one side, and on their respective expression in the heterogeneous cardiac tissue, on the other side. The aim of this review is to report and discuss the major works on IKs and IKr and the most recent ones that help to understand the precise function of these currents in the heart. PMID- 19683535 TI - Structural and functional characterization of the NHR1 domain of the Drosophila neuralized E3 ligase in the notch signaling pathway. AB - The Notch signaling pathway is critical for many developmental processes and requires complex trafficking of both Notch receptor and its ligands, Delta and Serrate. In Drosophila melanogaster, the endocytosis of Delta in the signal sending cell is essential for Notch receptor activation. The Neuralized protein from D. melanogaster (Neur) is a ubiquitin E3 ligase, which binds to Delta through its first neuralized homology repeat 1 (NHR1) domain and mediates the ubiquitination of Delta for endocytosis. Tom, a Bearded protein family member, inhibits the Neur-mediated endocytosis through interactions with the NHR1 domain. We have identified the domain boundaries of the novel NHR1 domain, using a screening system based on our cell-free protein synthesis method, and demonstrated that the identified Neur NHR1 domain had binding activity to the 20 residue peptide corresponding to motif 2 of Tom by isothermal titration calorimetry experiments. We also determined the solution structure of the Neur NHR1 domain by heteronuclear NMR methods, using a (15)N/(13)C-labeled sample. The Neur NHR1 domain adopts a characteristic beta-sandwich fold, consisting of a concave five-stranded antiparallel beta-sheet and a convex seven-stranded antiparallel beta-sheet. The long loop (L6) between the beta6 and beta7 strands covers the hydrophobic patch on the concave beta-sheet surface, and the Neur NHR1 domain forms a compact globular fold. Intriguingly, in spite of the slight, but distinct, differences in the topology of the secondary structure elements, the structure of the Neur NHR1 domain is quite similar to those of the B30.2/SPRY domains, which are known to mediate specific protein-protein interactions. Further NMR titration experiments of the Neur NHR1 domain with the 20-residue Tom peptide revealed that the resonances originating from the bottom area of the beta sandwich (the L3, L5, and L11 loops, as well as the tip of the L6 loop) were affected. In addition, a structural comparison of the Neur NHR1 domain with the first NHR domain of the human KIAA1787 protein, which is from another NHR subfamily and does not bind to the 20-residue Tom peptide, suggested the critical amino acid residues for the interactions between the Neur NHR1 domain and the Tom peptide. The present structural study will shed light on the role of the Neur NHR1 domain in the Notch signaling pathway. PMID- 19683533 TI - Cell therapy enhances function of remote non-infarcted myocardium. AB - Cell transplantation improves cardiac function after myocardial infarction; however, the underlying mechanisms are not well-understood. Therefore, the goals of this study were to determine if neonatal rat cardiomyocytes transplanted into adult rat hearts 1 week after infarction would, after 8-10 weeks: 1) improve global myocardial function, 2) contract in a Ca2+ dependent manner, 3) influence mechanical properties of remote uninjured myocardium and 4) alter passive mechanical properties of infarct regions. The cardiomyocytes formed small grafts of ultrastructurally maturing myocardium that enhanced fractional shortening compared to non-treated infarcted hearts. Chemically demembranated tissue strips of cardiomyocyte grafts produced force when activated by Ca2+, whereas scar tissue did not. Furthermore, the Ca2+ sensitivity of force was greater in cardiomyocyte grafts compared to control myocardium. Surprisingly, cardiomyocytes grafts isolated in the infarct zone increased Ca2+ sensitivity of remote uninjured myocardium to levels greater than either remote myocardium from non treated infarcted hearts or sham-operated controls. Enhanced calcium sensitivity was associated with decreased phosphorylation of cTnT, tropomyosin and MLC2, but not changes in myosin or troponin isoforms. Passive compliance of grafts resembled normal myocardium, while infarct tissue distant from grafts had compliance typical of scar. Thus, cardiomyocyte grafts are contractile, improve local tissue compliance and enhance calcium sensitivity of remote myocardium. Because the volume of remote myocardium greatly exceeds that of the grafts, this enhanced calcium sensitivity may be a major contributor to global improvements in ventricular function after cell transplantation. PMID- 19683536 TI - Cell cycle regulatory effects of retinoic Acid and forskolin are mediated by the cyclin C gene. AB - As a partner of cyclin-dependent kinase (CDK) 3, Cyclin C controls cellular proliferation and, together with CDK8, represses gene transcription. In this study, we showed that the highly expressed Cyclin C gene is a direct target of the nuclear hormone all-trans retinoic acid (RA) in HEK293 human embryonal kidney cells. The RA receptor (RAR) gamma associates with a Cyclin C promoter region containing two RAR binding sites. The Cyclin C gene also directly responds to the cAMP activator Forskolin via the transcription factor CREB1 (cAMP response element-binding protein 1), for which we identified four binding sites within the first 2250 bp of its promoter. RARgamma and CREB1 show functional convergence via the corepressor NCoR1, which controls in particular the Forskolin response of Cyclin C. The histone deacetylases 1, 5, 6, 7 and 11 are involved in the basal expression of Cyclin C, but in HEK293 and MCF-7 human breast carcinoma cells the antiproliferative effects of the histone deacetylase inhibitor SAHA (suberoylanilide hydroxamic acid) are not mediated by Cyclin C. However, cell cycle progressing effects of all-trans RA and Forskolin are dependent on Cyclin C expression levels. This suggests that the primary regulation of Cyclin C by all trans RA and Forskolin mediates some of the cell cycle control actions of these compounds. PMID- 19683537 TI - Role of a conserved prolyl residue (Pro173) of MotA in the mechanochemical reaction cycle of the proton-driven flagellar motor of Salmonella. AB - The MotA/B complex acts as the stator of the proton-driven bacterial flagellar motor. Proton translocation through the stator complex is efficiently coupled with torque generation by the stator-rotor interactions. In Salmonella enterica serovar Typhimurium, the highly conserved Pro173 residue of MotA is close to the absolutely conserved Asp33 residue of MotB, which is believed to be a proton binding site. Pro173 is postulated to be involved in coupling proton influx to torque generation. However, it remains unknown what critical function Pro173 carries out. Here, we characterize the motility and the torque-speed relation of the flagellar motor of the slow motile motA(P173A) mutant of Salmonella. Stall torque produced by the mutant motor was at the wild-type level, indicating that neither the number of stators in the motor nor the rotor-stator interaction is affected by the P173A substitution. In agreement with this, the motA(P173A) allele exerted a strong dominant-negative effect on wild-type motility. In contrast, high-speed rotation at low load was significantly impaired by the mutation, suggesting that the maximum rate of torque generation cycle is severely limited. Simulation of the torque-speed curve by a simple kinetic model indicated that the mutation reduces the rate of conformational changes of the MotA/B complex that switches the exposure of Asp33 to the outside and the inside of the cell, thereby slowing down the mechanochemical reaction cycle. Based on these results, we propose that Pro173 plays an important role in facilitating the conformational dynamics of the stator complex for rapid proton translocation and torque generation cycle. PMID- 19683538 TI - The 1.9 a structure of human alpha-N-acetylgalactosaminidase: The molecular basis of Schindler and Kanzaki diseases. AB - alpha-N-acetylgalactosaminidase (alpha-NAGAL; E.C. 3.2.1.49) is a lysosomal exoglycosidase that cleaves terminal alpha-N-acetylgalactosamine residues from glycopeptides and glycolipids. In humans, a deficiency of alpha-NAGAL activity results in the lysosomal storage disorders Schindler disease and Kanzaki disease. To better understand the molecular defects in the diseases, we determined the crystal structure of human alpha-NAGAL after expressing wild-type and glycosylation-deficient glycoproteins in recombinant insect cell expression systems. We measured the enzymatic parameters of our purified wild-type and mutant enzymes, establishing their enzymatic equivalence. To investigate the binding specificity and catalytic mechanism of the human alpha-NAGAL enzyme, we determined three crystallographic complexes with different catalytic products bound in the active site of the enzyme. To better understand how individual defects in the alpha-NAGAL glycoprotein lead to Schindler disease, we analyzed the effect of disease-causing mutations on the three-dimensional structure. PMID- 19683539 TI - Structural and kinetic studies of the allosteric transition in Sulfolobus solfataricus uracil phosphoribosyltransferase: Permanent activation by engineering of the C-terminus. AB - Uracil phosphoribosyltransferase catalyzes the conversion of 5-phosphoribosyl alpha-1-diphosphate (PRPP) and uracil to uridine monophosphate (UMP) and diphosphate (PP(i)). The tetrameric enzyme from Sulfolobus solfataricus has a unique type of allosteric regulation by cytidine triphosphate (CTP) and guanosine triphosphate (GTP). Here we report two structures of the activated state in complex with GTP. One structure (refined at 2.8-A resolution) contains PRPP in all active sites, while the other structure (refined at 2.9-A resolution) has PRPP in two sites and the hydrolysis products, ribose-5-phosphate and PP(i), in the other sites. Combined with three existing structures of uracil phosphoribosyltransferase in complex with UMP and the allosteric inhibitor cytidine triphosphate (CTP), these structures provide valuable insight into the mechanism of allosteric transition from inhibited to active enzyme. The regulatory triphosphates bind at a site in the center of the tetramer in a different manner and change the quaternary arrangement. Both effectors contact Pro94 at the beginning of a long beta-strand in the dimer interface, which extends into a flexible loop over the active site. In the GTP-bound state, two flexible loop residues, Tyr123 and Lys125, bind the PP(i) moiety of PRPP in the neighboring subunit and contribute to catalysis, while in the inhibited state, they contribute to the configuration of the active site for UMP rather than PRPP binding. The C-terminal Gly216 participates in a hydrogen-bond network in the dimer interface that stabilizes the inhibited, but not the activated, state. Tagging the C-terminus with additional amino acids generates an endogenously activated enzyme that binds GTP without effects on activity. PMID- 19683541 TI - How to arm a supervillin: designing F-actin binding activity into supervillin headpiece. AB - Villin-type headpiece domains are compact motifs that have been used extensively as model systems for protein folding. Although the majority of headpiece domains bind actin, there are some that lack this activity. Here, we present the first NMR solution structure and (15)N-relaxation analysis of a villin-type headpiece domain natively devoid of F-actin binding activity, that of supervillin headpiece (SVHP). The structure was found to be similar to that of other headpiece domains that bind F-actin. Our NMR analysis demonstrates that SVHP lacks a conformationally flexible region (V-loop) present in all other villin-type headpiece domains and which is essential to the phosphoryl regulation of dematin headpiece. In comparing the electrostatic surface potential map of SVHP to that of other villin-type headpiece domains with significant affinity for F-actin, we identified a positive surface potential conserved among headpiece domains that bind F-actin but absent from SVHP. A single point mutation (L38K) in SVHP, which creates a similar positive surface potential, endowed SVHP with specific affinity for F-actin that is within an order of magnitude of the tightest binding headpiece domains. We propose that this effect is likely conferred by a specific buried salt bridge between headpiece and actin. As no high-resolution structural information exists for the villin-type headpiece F-actin complex, our results demonstrate that through positive mutagenesis, it is possible to design binding activity into homologous proteins without structural information of the counterpart's binding surface. PMID- 19683540 TI - Structural basis of HIV-1 activation by NF-kappaB--a higher-order complex of p50:RelA bound to the HIV-1 LTR. AB - The activation and latency of human immunodeficiency virus type 1 (HIV-1) are tightly controlled by the transcriptional activity of its long terminal repeat (LTR) region. The LTR is regulated by viral proteins as well as host factors, including the nuclear factor kappaB (NF-kappaB) that becomes activated in virus infected cells. The two tandem NF-kappaB sites of the LTR are among the most highly conserved sequence elements of the HIV-1 genome. Puzzlingly, these sites are arranged in a manner that seems to preclude simultaneous binding of both sites by NF-kappaB, although previous biochemical work suggests otherwise. Here, we have determined the crystal structure of p50:RelA bound to the tandem kappaB element of the HIV-1 LTR as a dimeric dimer, providing direct structural evidence that NF-kappaB can occupy both sites simultaneously. The two p50:RelA dimers bind the adjacent kappaB sites and interact through a protein contact that is accommodated by DNA bending. The two dimers clamp DNA from opposite faces of the double helix and form a topological trap of the bound DNA. Consistent with these structural features, our biochemical analyses indicate that p50:RelA binds the HIV-1 LTR tandem kappaB sites with an apparent anti-cooperativity but enhanced kinetic stability. The slow on and off rates we observe may be relevant to viral latency because viral activation requires sustained NF-kappaB activation. Furthermore, our work demonstrates that the specific arrangement of the two kappaB sites on the HIV-1 LTR can modulate the assembly kinetics of the higher order NF-kappaB complex on the viral promoter. This phenomenon is unlikely restricted to the HIV-1 LTR but probably represents a general mechanism for the function of composite DNA elements in transcription. PMID- 19683542 TI - Male gamblers have significantly greater salivary cortisol before and after betting on a horse race, than do female gamblers. AB - Prevalence rates of gambling are influenced by gender. Among normative populations, hypothalamic-pituitary-adrenal (HPA) axis response to stress is affected by gender. However, pathological, compared to recreational, gamblers demonstrate perturbations in HPA activation in response to gambling stimuli. We examined whether there were gender differences in HPA response to gambling in a naturalistic setting among horse-race bettors and scratch-off lottery bettors. Salivary cortisol was collected from horse-race gamblers (n=32) and scratch-off lottery ticket players (n=39) before and after (0, 10, or 20 min) betting on a horse race at an off-track betting establishment. Salivary cortisol levels were significantly higher among men than among women, both prior to and following, betting on a horse race. Among women, but not men, there was a decline in salivary cortisol across time in scratch-off bettors, whereas women horse-race bettors maintained consistent low concentrations of salivary cortisol at every time point sampled. Together these data suggest that engaging in gambling may have different effects on stress responses of men, compared to women. Whether these gender differences in HPA activation contribute to gender-related differences in gambling behavior is the subject of ongoing investigation. PMID- 19683543 TI - Effect of resistance exercise on muscle steroid receptor protein content in strength-trained men and women. AB - The purpose of this study was to examine the acute effect of resistance exercise (RE) on muscle androgen receptor (AR) and glucocorticoid receptor (GR) protein content. Fifteen resistance-trained men (n=8; 21+/-1 years, 175.3+/-6.7 cm, 90.8+/-11.6 kg) and women (n=7; 24+/-5 years, 164.6+/-6.7 cm, 76.4+/-15.6 kg) completed 6 sets of 10 repetitions of heavy squats. Blood samples were obtained before RE, after 3 and 6 sets of squats, and 5, 15, 30 and 70 min after RE. Muscle biopsies from the vastus lateralis were obtained before RE, and 10 min and 70 min after RE. Blood samples were analyzed for total and free testosterone concentrations and muscle samples were analyzed for AR and GR protein content. Circulating total testosterone increased significantly (p< or =0.05) in men and free testosterone increased in men and women with exercise. AR was significantly reduced at 70 min post-exercise in men and at 10 min post-exercise in women compared to pre-exercise. There were no changes in GR following RE, but GR was significantly higher in women compared to men. These findings support a current paradigm for stabilization followed by a reduction and then a rebound in the acute AR response to RE but demonstrate that gender differences exist in the timeline of the AR response. PMID- 19683544 TI - Stimulus luminance and the spatial acuity of domestic fowl (Gallus g. domesticus). AB - The luminance dependence of spatial acuity in domestic fowl was measured directly over stimulus luminances ranging from 0.06 to 57.35 cd m(-2). At the highest luminance, acuity was around 6.5 c deg(-1), in agreement with previous studies in this species. As stimulus luminance decreased, acuity fell with increasing rate to 3.2 c deg(-1) at 0.06 cd m(-2), following the same shape as acuity functions for other mammalian and avian species. These findings suggest that the rod-cone transition for domestic fowl is between 0.45 and 1.79 cd m(-2). Over the photopic range from 1.79 to 57.35 cd m(-2) the change of acuity for fowl was 1%, compared with 32% for humans. For domestic fowl, the Rovamo-Barten MTF model of contrast sensitivity accounted for the behaviour of acuity as a function of luminance down to mesopic levels. PMID- 19683545 TI - Recommendations for advancing opportunities to increase physical activity in racial/ethnic minority communities. AB - We provide suggestions for advancing opportunities for effective and sustainable strategies for increasing physical activity in racial/ethnic minority populations. PMID- 19683547 TI - Perseveration related to frontal lesion in mice using the olfactory H-maze. AB - The delayed reaction paradigm, consisting to discover two different rules consecutively (delayed alternation and non-alternation task) followed by a delayed reversal task, is a specific marker for the functioning of primate prefrontal cortex. Although several works in rodents report the use of operant delayed alternation tasks, in none of the studies mice with lesion of the prefrontal cortex were used in this paradigm. In the current study, mouse experiments were conducted using a new, totally automated device, the olfactory H maze. Here, we show that unilateral lesion of the dorsomedial prefrontal cortex in mice induced similar deficits to those observed after frontal lesions in monkeys and humans. These pronounced learning deficits seem to come from difficulty elaborating a new rule and the inability to inhibit the previous rule, characterized by perseveration after prefrontal cortex lesion. The present results demonstrate that this very simple experimental paradigm using the olfactory H-maze presents the advantage to be fast (one training session) and well suited to assess the frontal functions in mice. It should be useful for testing pharmacological or stem cell approaches in order to reduce organic damages or gain insight into the cognitive functions of the frontal cortex using transgenic or gene-targeting mice. PMID- 19683546 TI - Entry inhibitors in the treatment of HIV-1 infection. AB - Infection of target cells by HIV is a complex, multi-stage process involving attachment to host cells and CD4 binding, coreceptor binding, and membrane fusion. Drugs that block HIV entry are collectively known as entry inhibitors, but comprise a complex group of drugs with multiple mechanisms of action depending on the stage of the entry process at which they act. Two entry inhibitors, maraviroc and enfuvirtide, have been approved for the treatment of HIV-1 infection, and a number of agents are in development. This review covers the entry inhibitors and their use in the management of HIV-1 infection. This article forms part of a special issue of Antiviral Research marking the 25th anniversary of antiretroviral drug discovery and development, Vol 85, issue 1, 2010. PMID- 19683548 TI - Detection of colon flora in peritoneal drain fluid after colorectal surgery: can RT-PCR play a role in diagnosing anastomotic leakage? AB - A semi-quantitative Real-Time PCR strategy was developed to identify potential indicator organisms for anastomotic leakage in peritoneal drainage fluid, Escherichia coli and Enterococcus faecalis. The analytical performance of the amplification method was validated with 10 culture-positive and 7 culture negative peritoneal drain fluid samples, obtained from 9 different patients with a colorectal anastomosis. Real-Time PCR results were fully concordant with the microbiological culture results. However, among the culture-negative samples, four false-positive RT-PCR results were found. All false-positives originated from a single patient with a surgical site infection. This may indicate an elevated sensitivity of the RT-PCR method. The results showed that the semi quantitative RT-PCR method has a clear potential to be useful as a powerful tool in early detection of anastomotic leakage. PMID- 19683549 TI - Double tactile sensations evoked by a single visual stimulus on a rubber hand. AB - The classical rubber hand illusion involves individuals misattributing tactile sensations 'felt' by their real hand hidden from view to a rubber prosthetic hand that they 'see' being tactilely stimulated in synchrony. However, from previous studies, it is not clear whether individuals actually feel the tactile sensation in the rubber hand, real hand, or both because the same part of the rubber and real hands were stimulated simultaneously. Here, we attempted to isolate subjects' sensations attributed to the rubber hand from those sensed in the real hand by placing the rubber and real hands in opposing orientations (e.g., palm up versus dorsum up). Interestingly, half of the subjects reported two tactile sensations for one visual stimulus, that is, one in the rubber finger stimulated visually with a light source and one in the real finger overlapping the rubber finger. This finding suggests that the tactile sensation induced by the visual stimulus is referred to the rubber hand and real hand simultaneously. Thus, both visuo-spatial and somatic codes are used in the localization of tactile sensation in the rubber hand illusion. PMID- 19683550 TI - Development of a high-throughput method to evaluate the impact of inhibitory compounds from lignocellulosic hydrolysates on the growth of Zymomonas mobilis. AB - Overcoming the effects of hydrolysate toxicity towards ethanologens is a key technical barrier in the biochemical conversion process for biomass feedstocks to ethanol. Despite its importance, the complexity of the hydrolysate toxicity phenomena and the lack of systematic studies, analysis and tools surrounding this issue have blocked a full understanding of relationships involving toxic compounds in hydrolysates and their effects on ethanologen growth and fermentation. In this study, we developed a quantitative, high-throughput biological growth assay using an automated turbidometer to obtain detailed inhibitory kinetics for individual compounds present in lignocellulosic biomass hydrolysate. Information about prolonged lag time and final cell densities can also be obtained. The effects of furfural, hydroxymethylfurfural (HMF), acetate and ethanol on growth rate and final cell densities of Zymomonas mobilis 8b on glucose are presented. This method was also shown to be of value in toxicity studies of hydrolysate itself, despite the highly colored nature of this material. Using this approach, we can generate comprehensive inhibitory profiles with many individual compounds and develop models that predict and examine toxic effects in the complex mixture of hydrolysates, leading to the development of improved pretreatment and conditioning processes as well as fermentation organisms. PMID- 19683551 TI - Construction and characterization of centromeric, episomal and GFP-containing vectors for Saccharomyces cerevisiae prototrophic strains. AB - A set of shuttle yeast vectors containing the dominant selectable markers KanMX4 or HphMX4 cassettes, conferring resistance to geneticin and hygromycin B, respectively, was constructed. Dominant selectable markers are useful for genetic manipulation of natural, wine and industrial strains which do not contain any auxotrophic markers as well as of strains which cannot grow on synthetic mineral medium. Vectors were characterized by (i) copy number, (ii) mitotic stability both in selective and non-selective conditions, (iii) the efficiency and frequency of transformations, (iv) optimal adaptation times in non-selective media, (v) optimum conditions for transformation of various laboratory, commercial and wine strains, and (vi) expression level of an inserted gene. Furthermore we produced GFP-containing vectors that can be used for protein subcellular localization in prototrophic strains. PMID- 19683552 TI - Intracellular trafficking of cell-penetrating peptide-avidin complexes. PMID- 19683553 TI - Synthesis and physicochemical characterization of a novel amphiphilic polylactic acid-hyperbranched polyglycerol conjugate for protein delivery. AB - Amphiphilic copolymers with polylactic acid (PLA) chains grafted onto hyperbranched polyglycerol (HPG) have been synthesized and characterized. The copolymer nanoparticles with corona and core structure were formed by self assembly in aqueous solution. The loading capacity and association efficiency were up to 23% and 86%, respectively. Protein release profiles with different copolymer compositions and BSA concentrations all showed a burst effect followed by a continuous release phase. The released BSA from the copolymer nanoparticles remained in its original structure over a period of 4 days, as testified by circular dichroism spectroscopy. Furthermore, cell viability research suggested good biocompatibility of the copolymer nanoparticles, which have a promising potential for protein delivery system. PMID- 19683554 TI - Purification and molecular cloning of antimicrobial peptides from Scots pine seedlings. AB - A novel protocol for rapid and efficient purification of antimicrobial peptides from plant seedlings has been developed. Two peptides with antimicrobial activity, designated p1 and p2, were purified nearly to homogeneity from Scots pine seedlings by a combination of sulfuric acid extraction, ammonium sulfate precipitation, heat-inactivation and ion-exchange chromatography on phosphocellulose. Purified proteins had molecular masses of 11 kDa (p1) and 5.8 kDa (p2) and were identified by mass spectrometry as defensin and lipid-transfer protein, respectively. We demonstrated their growth inhibitory effects against a group of phytopathogenic fungi. Furthermore, we report for the first time molecular cloning and characterization of defensin 1 cDNA from Scots pine. A cDNA expression library from 7 days Scots pine seedlings was generated and used to isolate a cDNA clone corresponding to Scots pine defensin, termed PsDef1. The full-length coding sequence of PsDef1 is 252 bp in length and has an open reading frame capable to encode a protein of 83 amino residues. The deduced sequence has the typical features of plant defensins, including an endoplasmic reticulum signal sequence of 33aa, followed by a characteristic defensin domain of 50 amino acids representing its active form. The calculated molecular weight of the mature form of PsDef1 is 5601.6 Da, which correlates well with the results of SDS-PAGE analysis. Finally, the antimicrobial properties of PsDef1 against a panel of fungi and bacteria define it as a member of the morphogenic group of plant defensins. PMID- 19683555 TI - Role of human leukocyte antigen, killer-cell immunoglobulin-like receptors, and cytokine gene polymorphisms in leptospirosis. AB - Leptospirosis is an emerging zoonotic disease caused by pathogenic species of the genus Leptospira. It has a broad range of clinical presentations in humans. Although progress has been made in the characterization of the host immune system factors that may affect disease progression and outcome, to date few reports have addressed the role of genetic polymorphisms in the susceptibility to leptospirosis. In this work a group of patients with a history of leptospiral infection and a control group were compared for polymorphisms in the human leukocyte antigen (HLA), in killer-cell immunoglobulin-like receptors (KIR), and in cytokine genes. Alleles in the HLA-A and -B loci were associated with susceptibility, as were the class I haplotype A*01-B*08-Cw*07 and the 8.1 ancestral haplotype (A*01-B*08-Cw*07-DRB1*03-DQB1*02). Single nucleotide polymorphisms in the interleukin (IL)-4 and IL-4Ralpha genes also had significantly higher frequencies in the patient group. No association was reported between KIR gene profile and leptospirosis. This work highlights the importance of using genetic polymorphisms to better understand the mechanisms involved in the immune response to leptospirosis. PMID- 19683556 TI - High-resolution ERP mapping of cortical activation related to implicit object location memory. AB - High-density event-related potentials (ERPs) were recorded during an object recognition task which involved task-irrelevant changes in the location of studied objects. Participants categorised objects as studied or novel while data were analysed to ascertain the effect of the location changes on performance and waveform topography. Our results indicate that humans can classify objects faster and more accurately when using implicit spatial memory. Individual differences observed in object recognition proficiency were absent if objects were presented in their 'correct' location. In a second experiment we replicated the behavioural findings while manipulating viewpoint to discount scene recognition as an underlying factor. We propose a model which includes activation of the right medial temporal lobe prior to P300 elicitation to account for the prophylactic effect of implicit processing on object recognition. Hemispheric differences in parietal componentry dependant on sex of participant were also observed and are discussed in relation to differential strategies. PMID- 19683557 TI - Potential role of estrogen in regulation of the insulin-like growth factor2-H19 locus in the rat testis. AB - The selective estrogen receptor modulator, tamoxifen, has been shown to reduce DNA methylation at Insulin-like growth factor 2/H19 differentially methylated region (Igf2/H19 DMR) in the spermatozoa of the Holtzman rats. Since imprint at this locus is acquired during spermatogenesis in the male germ-line, we hypothesized role for estrogen signaling in the methylation dynamics in the testis. The present study was designed to identify putative estrogen response elements (ERE) at Igf2/H19 DMR and their interaction with DNA methylation pathway. Here, we demonstrate presence of functional ERE at 2637/2655 base pair on Igf2/H19 DMR in testicular germ cells, which was found to bind to estrogen receptor beta (ER beta) in the chromatin immunoprecipitation assay. Tamoxifen attenuated ER beta-ERE association thereby acting as an estrogen antagonist at this locus. Further mechanistic study involving colocalization and immunoprecipitation assay revealed interaction of ER beta and Dnmt1 in the testis. The study provides evidence for the role for estrogen in acquisition of imprint at Igf2/H19 DMR in testis and help in understanding molecular mechanism of environmental estrogens impacting male fertility. PMID- 19683558 TI - The elevator illusion results from the combination of body orientation and egocentric perception. AB - Perception of body orientation and apparent location of objects are altered when humans are using assisted means of locomotion and the resultant of the imposed acceleration and gravity is no longer aligned with the gravitational vertical. As the otolithic system cannot discriminate the acceleration of gravity from sustained inertial accelerations, individuals would perceive the resultant acceleration vector (GiA) as the vertical. However, when subjects are aligned on the GiA, an increase in the magnitude of GiA induced a lowering of the apparent visual horizon (i.e. "elevator illusion"). The main aim of this study was to quantify the contribution of body and egocentric perception in the elevator illusion. While being exposed to 1G and 1.3G and aligned on the GiA acceleration, subjects (N=20) were asked (1) to set a luminous target to the subjective horizon, (2) to set a luminous target on "straight ahead" position (egocentric task) and (3) to rotate a tilting tube to their subjective perception of body orientation. Results showed that increasing GiA lowered horizon and egocentric settings and induces a backward body tilt perception. Moreover, the elevator illusion can be expressed as the additive combination of two processes: one that is dependent on body tilt perception, and the other that is dependent on egocentric perception. Both misperceptions in hypergravity may be considered to be a consequence of excessive shearing of the otolith organs. However large inter individual differences in body tilt perception were observed. This last result was discussed in terms of the contribution of extravestibular graviceptors. PMID- 19683560 TI - Metastasis mechanisms. AB - Metastasis, the spread of malignant cells from a primary tumor to distant sites, poses the biggest problem to cancer treatment and is the main cause of death of cancer patients. It occurs in a series of discrete steps, which have been modeled into a "metastatic cascade". In this review, we comprehensively describe the molecular and cellular mechanisms underlying the different steps, including Epithelial-Mesenchymal Transition (EMT), invasion, anoikis, angiogenesis, transport through vessels and outgrowth of secondary tumors. Furthermore, we implement recent findings that have broadened and challenged the classical view on the metastatic cascade, for example the establishment of a "premetastatic niche", the requirement of stem cell-like properties, the role of the tumor stroma and paracrine interactions of the tumor with cells in distant anatomical sites. A better understanding of the molecular processes underlying metastasis will conceivably present us with novel targets for therapeutic intervention. PMID- 19683559 TI - Trop2: a possible therapeutic target for late stage epithelial carcinomas. AB - Trop2 is a cell-surface glycoprotein overexpressed in a variety of late stage epithelial carcinomas with low to no expression in normal tissues. Some of the important roles that Trop2 plays in epithelial cancers have recently been revealed. Trop2 overexpression is associated with decreased patient survival as well as increased tumor aggressiveness and metastasis. Its overexpression in metastatic tissue makes it a very attractive and potential therapeutic target for late stage disease. This protein can transduce an intracellular calcium signal and contains a conserved phosphatidylinositol 4,5-bisphosphate (PIP(2)) binding motif as well as a serine phosphorylation site which interacts with protein kinase C. This protein has recently being found to be expressed in cells with stem-like properties which allude to a potential role in progenitor cell biology. Further understanding of the signaling pathways involved with this molecule and its important role in metastasis will shed new light on the mechanism of Trop2 overexpression in late stage disease and could result in the development of new therapies targeting this widely overexpressed oncogene. PMID- 19683561 TI - Sciatic nerve transection increases gluthatione antioxidant system activity and neuronal nitric oxide synthase expression in the spinal cord. AB - Glutathione (GSH) is a major non-enzymatic antioxidant which is present in all tissues. Its protective actions occur through different pathways such its role as a substrate of antioxidant enzymes, such as glutathione peroxidase (GPx) and glutathione-S-transferase (GST). Nitric oxide (NO) is involved in many physiological processes in the central nervous system, including nociception. In spite of much evidence concerning oxidative and nitrosative stress and neuropathic pain, the exact role of these molecules in pain processing is still unknown. Sciatic nerve transection (SNT) was employed to induce neuropathic pain in rats. Glutathione peroxidase (GPx) and glutathione-S-transferase (GST) activities, glutathione (GSH) content, GSH/GSSG ratio, nitric oxide metabolites (NOx) and neuronal nitric oxide synthase (nNOS) protein expression in the lumbosacral spinal cord were determined. All of these analyses were performed in the SNT and sham groups 1, 3, 7 and 15 days after surgery. There was an increase in GPx activity and in GSH content 3 days after surgery in both sham and SNT groups, but the GSH/GSSG ratio increased only in the SNT group in this time point. nNOS expression was upregulated 7 days post SNT. NOx was detected 1 day after surgery in sham and SNT groups, but at 7 and 15 days, the increase occurred only in SNT animals. These results support the role of the gluthatione system in pain physiology and highlight the involvement of NO as an important molecule related to nociception. PMID- 19683563 TI - miR-34a, a microRNA up-regulated in a double transgenic mouse model of Alzheimer's disease, inhibits bcl2 translation. AB - MicroRNAs (miRNAs) are short noncoding regulatory RNA molecules that modulate protein expression by inhibiting mRNA translation or promoting mRNA degradation. However, little is understood about the roles of miRNAs in Alzheimer's disease. During a research for miRNAs that are differentially expressed in cerebral cortex of APPswe/PSDeltaE9 mice (a model for Alzheimer's disease) and age-matched controls, one candidate miRNA that is relatively highly expressed, miR-34a, was studied further because sequence analysis suggested a likely interaction with the 3'-untranslated region of bcl2 mRNA. We show that the expression of miR-34a is inversely correlated with the protein level of bcl2 in APPswe/PSDeltaE9 mice and age-matched controls, and miR-34a expression directly inhibits bcl2 translation in SH-SY5Y cells. No effect on bcl2 mRNA level was observed. Western blot analysis of active caspase-3 showed higher levels in APPswe/PSDeltaE9 mice and stable transfecant cell line of miR-34a than in controls. Consistently, miR-34a knockdown through antisense LNA oligonucleotides increased the level of bcl2 protein in SH-SY5Y cells, which was accompanied by a decrease of active caspase 3. These findings suggested that bcl2 is an important functional target for miR 34a, and the abnormal expression of miR-34a may contribute to the pathogenesis of Alzheimer's disease, at least in part by affecting the expression of bcl2. PMID- 19683564 TI - In silico predication of nuclear hormone receptors for organic pollutants by homology modeling and molecular docking. AB - Homology modeling and molecular docking were used to in silico predict the rat nuclear hormone receptors of different organic pollutants. Rat aryl hydrocarbon receptor (rAhR), constitutive androstane receptor (rCAR) and pregnane X receptor (rPXR) were chosen as the target nuclear receptors. 3D models of ligand binding domains of rAhR, rCAR and rPXR were constructed by MODELLER 9V6 and assessed by the Procheck and Prosa 2003. Surflex-Dock program was applied to bind the different organic pollutants into the three receptors to predict their affinities. The results of docking experiments demonstrated that three polybrominated dibenzofurans (PBDFs, including TretaBDF, PentaBDF and HexaBDF) and 3,3',4,4',5'-pentachlorobiphenyl (PCB126) would be better categorized by rAhR dependent mechanism, but four polybrominated diphenyl ethers (PBDEs, including BDE47, BDE80, BDE99 and BDE153) and 2,2',4,4',5,5'-hexachlorobiphenyl (PCB153) by rCAR and rPXR-dependent mechanism. For benzo(a)pyrene and pyrene, they have high affinities with the three target receptors, which suggests that "crosstalk" among the receptors might occur during the receptor induction. The results of this study are consistent with those of animal experiments reported by previous literatures, which suggest that homology modeling and molecular docking would have the potential to predict the nuclear hormone receptors of environmental pollutants. PMID- 19683565 TI - Influence of strain and sex on the metabolic profile of rats in repeated dose toxicological studies. AB - The impact of the strain on the metabolite profile of plasma samples in rats dosed with 2500 ppm 2-methyl-4-chlorophenoxyacetic acid (MCPA acid) or 45 mg/kg bw/day 4-chloro-3-nitroaniline (4C3N) for 4 weeks was evaluated. Four different strains were used: two Wistar strains (Crl:WI(Han), Han:RCC:WIST(SPF)), one Sprague-Dawley (Crl:CD) and one Fisher strain (F-344/Crl). The metabolite profiles in the plasma were measured by LC-MS and GC-MS. The profound changes of the metabolite values induced by the MCPA acid treatment outweighed slight deviations caused by physiological variations between the different rat strains. The metabolome changes of the MCPA acid in all strains could be related to toxicological "mode of action" patterns (peroxisome proliferator, renal organic anionic transporter inhibition) with Crl:WI(Han) rats as reference strain. 4C3N administration led to extravascular hemolytic anemia with a small number of metabolome changes, which were strain dependent. The metabolome pattern associated with "hemolytic anemia" established with the reference strain (Crl:Wi(Han)) was not sufficiently similar in other strains. Thus, comparable metabolome profiles were obtained in different rat strains for a compound inducing profound metabolite changes. For a compound with a weak profile the results were more variable and appeared to be strain dependent. PMID- 19683566 TI - Lactational fenvalerate exposure permanently impairs testicular development and spermatogenesis in mice. AB - Fenvalerate, a widely used synthetic pyrethroid insecticide, has been associated with poor semen quality in human being. However, little is known about the effects of lactational fenvalerate exposure on testicular development and spermatogenesis. The purpose of the present study was to investigate the effects of maternal fenvalerate exposure during lactation on testicular development and spermatogenesis in male offspring. Maternal mice were administered with fenvalerate (60 mg/kg) by gavage daily from postnatal day (PND) 0 to PND21. Lactational fenvalerate exposure markedly decreased the absolute and relative weights of testes and increased the number of apoptotic cells in testes of pups at weaning. Histological examinations showed abnormal seminiferous tubules with large vacuoles or complete spermatogenic failure in testes of fenvalerate-treated mice at weaning. Additional experiment showed that lactational fenvalerate exposure markedly reduced mRNA and protein levels of testicular P450scc, a testosterone (T) synthesis enzyme. Consistent with down-regulation of testicular P450scc, the level of serum and testicular T at weaning was significantly decreased in pups whose mothers were exposed to fenvalerate during lactation. Although the expression of testicular P450scc and serum and testicular T in adulthood restored to control level, the decreased weight of testes and histological changes were irreversible. Importantly, the percentage of mature seminiferous tubules (stages VII and VIII) and the number of spermatozoa were obviously decreased in adult male mice whose mothers were exposed to fenvalerate during lactation. Taken together, these results suggest that lactational fenvalerate exposure permanently impairs testicular development and spermatogenesis. PMID- 19683562 TI - Diabetic keratopathy and treatment by modulation of the opioid growth factor (OGF)-OGF receptor (OGFr) axis with naltrexone: a review. AB - The opioid growth factor (OGF)-OGF receptors (OGFr) axis plays an important role in the homeostasis and re-epithelialization of the mammalian cornea. This tonically active growth regulatory inhibitory pathway is involved in cell replication, and the endogenous neuropeptide OGF targets cyclin-dependent kinase inhibitors, p16 and/or p21. Blockade of OGF-OGFr interfacing by systemic or topical administration of opioid antagonists such as naltrexone (NTX) results in accelerated DNA synthesis, cell replication, and tissue repair. Molecular manipulation of OGFr using sense constructs delayed corneal re-epithelialization, whereas antisense constructs accelerated repair of the corneal surface. Corneal keratopathy, a significant complication of diabetes mellitus, is manifested by delays in corneal re-epithelialization following surgery, injury, or disease. Tissue culture studies have shown that addition of NTX stimulates DNA synthesis and explant outgrowth of rabbit corneal epithelium, whereas OGF depresses DNA synthesis and explant outgrowth in a receptor-mediated manner. NTX accelerated corneal re-epithelialization in organ cultures of human and rabbit cornea. Systemic application of NTX to the abraded corneas of rats, and topical administration of NTX to the injured rabbit ocular surface, increased re epithelialization. Systemic injections or topical administration of NTX facilitates re-epithelialization of the cornea in diabetic rats. Given the vital role of the corneal epithelium in maintaining vision, the frequency of corneal complications related to diabetes (diabetic keratopathy), and the problems occurring in diabetic individuals postoperatively (e.g., vitrectomy), and that conventional therapies such as artificial tears and bandage contact lenses often fail, topical application of NTX merits clinical consideration. PMID- 19683567 TI - Diesel exhaust exposure induces angiogenesis. AB - Our aim was to test the hypothesis that exposure to whole diesel exhaust (WDE) would enhance angiogenesis/vasculogenesis. Male apolipoprotein E-deficient mice, with either scaffold implantation subcutaneously or hindlimb ischemia, were exposed to either WDE (containing diesel exhaust particle [DEP] at a concentration of about 1mg/m(3)) or filtered air 6 h/day, 5 days/week in a whole body exposure chamber for 2, 5, or 8 weeks, respectively. WDE exposure significantly increased total cell counts in the scaffolds, aortic, and perivascular fat tissues. Macrophage infiltration was enhanced and CD31 expression increased in the scaffolds, which was coupled by increased alpha smooth muscle actin (alpha-SMA) expression. WDE exposure led to increased CD31 expression, while decreasing endothelial nitric oxide synthase in the aortic wall. The vessel volume measured by micro-CT was increased in ischemic and non ischemic hindlimbs in response to WDE exposure. DEP exposure induced capillary like tube formation in endothelial cells in vitro, and caused capillary sprouting from aortic rings ex vivo. In addition, WDE exposure significantly increased mRNA expression of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor (HIF)-1alpha, while decreasing prolylhydroxylase (PHD) 2 expression. WDE exposure increases inflammatory cell infiltration, enhances the vessel volume/flow, and increases capillary tube formation and sprouting, thereby inducing angiogenesis and vasculogenesis. The angiogenic effects may occur through increasing HIF-1alpha and VEGF while decreasing PHD2 expression. PMID- 19683569 TI - Downstream mechanisms triggered by mitochondrial dysfunction in the basal ganglia: from experimental models to neurodegenerative diseases. AB - Mitochondrial dysfunctions have been implicated in the cellular processes underlying several neurodegenerative disorders affecting the basal ganglia. These include Huntington's chorea and Parkinson's disease, two highly debilitating motor disorders for which recent research has also involved gene mutation linked to mitochondrial deficits. Experimental models of basal ganglia diseases have been developed by using toxins able to disrupt mitochondrial function: these molecules act by selectively inhibiting mitochondrial respiratory complexes, uncoupling cellular respiration. This in turn leads to oxidative stress and energy deficit that trigger critical downstream mechanisms, ultimately resulting in neuronal vulnerability and loss. Here we review the molecular and cellular downstream effects triggered by mitochondrial dysfunction, and the different experimental models that are obtained by the administration of selective mitochondrial toxins or by the expression of mutant genes. PMID- 19683570 TI - Influence of degree of substitution of HES-HEMA on the release of incorporated drug models from corresponding hydrogels. AB - Hydrogel microparticles were produced by a radical polymerization of hydroxyethyl methacrylate-hydroxyethyl starch (HES-HEMA) in an all aqueous two-phase system (ATPS). The microspheres show a monomodal size distribution and have the ability to entrap high amounts of water. The release of proteins or other testing substances from the HES-HEMA hydrogels can be controlled by the choice of the network density of the hydrogel by varying the degree of substitution (DS), the size of the entrapped substance, and by conditions enhancing the degradation of the hydrogel network. PMID- 19683568 TI - Are there bi-directional associations between depressive symptoms and C-reactive protein in mid-life women? AB - OBJECTIVE: To test whether depressive symptoms are related to subsequent C reactive protein (CRP) levels and/or whether CRP levels are related to subsequent depressive symptoms in mid-life women. METHODS: Women enrolled in the Study of Women's Health Across the Nation (SWAN) were followed for 7years and had measures of CES-Depression scores and CRP seven times during the follow-up period. Women were pre- or early peri-menopausal at study entry and were of Caucasian, African American, Hispanic, Japanese, or Chinese race/ethnicity. Analyses were restricted to initially healthy women. RESULTS: Longitudinal mixed linear regression models adjusting for age, race, site, time between exams, and outcome variable at year X showed that higher CES-D scores predicted higher subsequent CRP levels and vice versa over a 7-year period. Full multivariate models adjusting for body mass index, physical activity, medications, health conditions, and other covariates showed that higher CRP levels at year X predicted higher CES-D scores at year X+1, p=0.03. Higher depressive symptoms predicted higher subsequent CRP levels at marginally significant levels, p=0.10. CONCLUSIONS: Higher CRP levels led to higher subsequent depressive symptoms, albeit the effect was small. The study demonstrates the importance of considering bi-directional relationships for depression and other psychosocial factors and risk for heart disease. PMID- 19683572 TI - In vitro receptivity of carbonate rocks to endolithic lichen-forming aposymbionts. AB - Sterile cultured isolates of lichen-forming aposymbionts have not yet been used to investigate lichen-rock interactions under controlled conditions. In this study mycobionts and photobiont of the endolithic lichens Bagliettoa baldensis and Bagliettoa marmorea were isolated and inoculated with coupons of one limestone and four marbles commonly employed in the Cultural Heritage framework. After one year of incubation, microscopic observations of polished cross-sections were performed to verify if the typical colonization patterns observed in the field may be reproduced in vitro and to evaluate the receptivity of the five lithotypes to endolithic lichens. The mycobionts of the two species developed both on the surface of and within all the lithotypes, showing different penetration pathways which depend on mineralogical and structural features and highlight different receptivity. By contrast, algae inoculated with the coupons did not penetrate them. Observations suggest that the hyphal penetration along intrinsic discontinuities of rocks is a relatively fast phenomenon when these organisms are generally considered as slow-growing. Samples from limestone outcrops and abandoned marble quarries, colonized by the same species or other representatives of Verrucariaceae, showed penetration pathways intriguingly similar to those reproduced in vitro and highlighted that lichen-driven erosion processes only increase the availability of hyphal passageways after a long-term colonization. These results show that in vitro incubation of sterile cultured lichen-forming ascomycetes with rock coupons is a practicable experimental system to investigate the lichen-rock interactions under controlled conditions and, together with analysis in situ, may support decisions on conservative treatments of historical and cultural significant stone substrata. PMID- 19683574 TI - Differential proteoglycan expression in two spinal cord regions after dorsal root injury. AB - Dorsal root injury leads to reactive gliosis in the spinal cord dorsal root entry zone and dorsal column, two regions that undergo Wallerian degeneration, but have distinct growth-inhibitory properties. This disparity could in part be due to differences in the number of degenerating sensory fibers, differences in glial cell activation, and/or to differential expression of growth-inhibitory molecules such as chondroitin sulfate proteoglycans. Laser capture microdissection of these two spinal cord white matter regions, followed by quantitative analysis of mRNA expression by real-time PCR, revealed that glial marker transcripts were differentially expressed post-injury and that the chondroitin sulfate proteoglycans Brevican and Versican V1 and V2 were preferentially up-regulated in the dorsal root entry zone, but not the dorsal column. These results indicate that reactive gliosis differs between these two regions and that Brevican and Versican are potential key molecules participating in the highly inhibitory properties of the dorsal root entry zone. PMID- 19683573 TI - Axonal and neuromuscular synaptic phenotypes in Wld(S), SOD1(G93A) and ostes mutant mice identified by fiber-optic confocal microendoscopy. AB - We used live imaging by fiber-optic confocal microendoscopy (CME) of yellow fluorescent protein (YFP) expression in motor neurons to observe and monitor axonal and neuromuscular synaptic phenotypes in mutant mice. First, we visualized slow degeneration of axons and motor nerve terminals at neuromuscular junctions following sciatic nerve injury in Wld(S) mice with slow Wallerian degeneration. Protection of axotomized motor nerve terminals was much weaker in Wld(S) heterozygotes than in homozygotes. We then induced covert modifiers of axonal and synaptic degeneration in heterozygous Wld(S) mice, by N-ethyl-N-nitrosourea (ENU) mutagenesis, and used CME to identify candidate mutants that either enhanced or suppressed axonal or synaptic degeneration. From 219 of the F1 progeny of ENU mutagenized BALB/c mice and thy1.2-YFP16/Wld(S) mice, CME revealed six phenodeviants with suppression of synaptic degeneration. Inheritance of synaptic protection was confirmed in three of these founders, with evidence of Mendelian inheritance of a dominant mutation in one of them (designated CEMOP_S5). We next applied CME repeatedly to living Wld(S) mice and to SOD1(G93A) mice, an animal model of motor neuron disease, and observed degeneration of identified neuromuscular synapses over a 1-4day period in both of these mutant lines. Finally, we used CME to observe slow axonal regeneration in the ENU-mutant ostes mouse strain. The data show that CME can be used to monitor covert axonal and neuromuscular synaptic pathology and, when combined with mutagenesis, to identify genetic modifiers of its progression in vivo. PMID- 19683575 TI - Role of plasminogen activation in neuronal organization and survival. AB - We characterized the interactions between plasminogen and neurons and investigated the associated effects on extracellular matrix proteolysis, cell morphology, adhesion, signaling and survival. Upon binding of plasminogen to neurons, the plasmin formed by constitutively expressed tissue plasminogen activator (tPA) degrades extracellular matrix proteins, leading to retraction of the neuron monolayer that detaches from the matrix. This sequence of events required both interaction of plasminogen with carboxy-terminal lysine residues and the proteolytic activity of plasmin. Surprisingly, 24h after plasminogen addition, plasmin-detached neurons survived and remained associated in clusters maintaining focal adhesion kinase phosphorylation contrasting with other adherent cell types fully dissociated by plasmin. However, long-term incubation (72 h) with plasminogen was associated with an increased rate of apoptosis, suggesting that prolonged exposure to plasmin may cause neurotoxicity. Regulation of neuronal organization and survival by plasminogen may be of pathophysiological relevance, as plasminogen is expressed in the brain and/or extravasate during vascular accidents or inflammatory processes. PMID- 19683576 TI - Interaction between DLX2 and EGFR regulates proliferation and neurogenesis of SVZ precursors. AB - In the postnatal subventricular zone (SVZ) neural stem cells (NSCs) give rise to transit-amplifying precursors (TAPs) expressing high levels of epidermal growth factor receptor (EGFR) that in turn generate neuroblasts. Both TAPs and neuroblasts express distal-less (DLX)2 homeobox transcription factor but the latter proliferate less. Modulation of its expression in vivo has revealed that DLX2 affects both neurogenesis and proliferation in the postnatal SVZ. However, the mechanisms underlying these effects are not clear. To investigate this issue we have here forced the expression of DLX2 in SVZ isolated NSCs growing in defined in vitro conditions. This analysis revealed that DLX2 affects the proliferation of SVZ precursors by regulating two distinct steps of neural lineage progression. Firstly, it promotes the lineage transition from NSCs to TAPs. Secondly it enhances the proliferative response of neuronal progenitors to EGF. Thus DLX2 and EGFR signalling interact at multiple levels to coordinate proliferation in the postnatal SVZ. PMID- 19683577 TI - Sprouty2 down-regulation promotes axon growth by adult sensory neurons. AB - Fibroblast growth factors (FGFs) play a prominent role in axonal growth during development and repair. Treatment with FGF-2 or overexpression of FGF receptors promotes peripheral axon regeneration mainly by activation of extracellular signal-regulated kinase (ERK). The Ras/Raf/ERK pathway is under the control of Sprouty proteins acting as negative feedback inhibitors. We investigated the expression of Sprouty isoforms in adult sensory neurons of dorsal root ganglia (DRG) as well as the effects of Sprouty inhibition on axon growth by small interfering RNAs (siRNAs). Sprouty2 revealed the highest expression level in DRG neurons. Down-regulation of Sprouty2 promoted elongative axon growth by adult sensory neurons accompanied by enhanced FGF-2-induced activation of ERK and Ras, whereas Sprouty2 overexpression inhibited axon growth. Sprouty2 was not regulated in vivo in response to a sciatic nerve lesion. Together, our results imply that Sprouty2 is highly expressed in adult peripheral neurons and its down-regulation strongly promotes elongative axon growth by activation of the Ras/Raf/ERK pathway. PMID- 19683578 TI - Bio-sintering processes in hexactinellid sponges: fusion of bio-silica in giant basal spicules from Monorhaphis chuni. AB - The two sponge classes, Hexactinellida and Demospongiae, comprise a skeleton that is composed of siliceous skeletal elements (spicules). Spicule growth proceeds by appositional layering of lamellae that consist of silica nanoparticles, which are synthesized via the sponge-specific enzyme silicatein. While in demosponges during maturation the lamellae consolidate to a solid rod, the lamellar organization of hexactinellid spicules largely persists. However, the innermost lamellae, near the spicule core, can also fuse to a solid axial cylinder. Similar to the fusion of siliceous nanoparticles and lamella, in several hexactinellid species individual spicules unify during sintering-like processes. Here, we study the different stages of a process that we termed bio-sintering, within the giant basal spicule (GBS) of Monorhaphis chuni. During this study, a major GBS protein component (27 kDa) was isolated and analyzed by MALDI-TOF-MS. The sequences were used to isolate and clone the encoding cDNA via degenerate primer PCR. Bioinformatic analyses revealed a significant sequence homology to silicatein. In addition, the native GBS protein was able to mediate bio-silica synthesis in vitro. We conclude that the syntheses of bio-silica in M. chuni, and the subsequent fusion of nanoparticles to lamellae, and finally to spicules, are enzymatically-driven by a silicatein-like protein. In addition, evidence is now presented that in hexactinellids those fusions involve sintering-like processes. PMID- 19683579 TI - Monolithic microfluidic mixing-spraying devices for time-resolved cryo-electron microscopy. AB - The goal of time-resolved cryo-electron microscopy is to determine structural models for transient functional states of large macromolecular complexes such as ribosomes and viruses. The challenge of time-resolved cryo-electron microscopy is to rapidly mix reactants, and then, following a defined time interval, to rapidly deposit them as a thin film and freeze the sample to the vitreous state. Here we describe a methodology in which reaction components are mixed and allowed to react, and are then sprayed onto an EM grid as it is being plunged into cryogen. All steps are accomplished by a monolithic, microfabricated silicon device that incorporates a mixer, reaction channel, and pneumatic sprayer in a single chip. We have found that microdroplets produced by air atomization spread to sufficiently thin films on a millisecond time scale provided that the carbon supporting film is made suitably hydrophilic. The device incorporates two T mixers flowing into a single channel of four butterfly-shaped mixing elements that ensure effective mixing, followed by a microfluidic reaction channel whose length can be varied to achieve the desired reaction time. The reaction channel is flanked by two ports connected to compressed humidified nitrogen gas (at 50 psi) to generate the spray. The monolithic mixer-sprayer is incorporated into a computer-controlled plunging apparatus. To test the mixing performance and the suitability of the device for preparation of biological macromolecules for cryo EM, ribosomes and ferritin were mixed in the device and sprayed onto grids. Three dimensional reconstructions of the ribosomes demonstrated retention of native structure, and 30S and 50S subunits were shown to be capable of reassociation into ribosomes after passage through the device. PMID- 19683580 TI - Automatic extraction of sulcal lines on cortical surfaces based on anisotropic geodesic distance. AB - Analyzing cortical sulci is important for studying cortical morphology and brain functions. Although sulcal lines on cortical surfaces can be defined in various ways, it is critical in a neuroimaging study to define a sulcal line along the valley of a cortical surface with a high curvature within a sulcus. To extract the sulcal lines automatically, we present a new geometric algorithm based on the computation of anisotropic skeletons of sulcal regions. Because anisotropic skeletons are highly adaptive to the anisotropic nature of the surface shape, the resulting sulcal lines lie accurately on the valleys of the sulcal areas. Our sulcal lines remain unchanged under local shape variabilities in different human brains. Through experiments, we show that the errors of the sulcal lines for both synthetic data and real cortical surfaces were nearly as constant as the function of random noise. By measuring the changes in sulcal shape in Alzheimer's disease (AD) patients, we further investigated the effectiveness of the accuracy of our sulcal lines using a large sample of MRI data. This study involved 70 normal controls (n [men/women]: 29/41, age [mean+/-SD]: 71.7+/-4.9 years), and 100 AD subjects (37/63, 72.3+/-5.5). We observe significantly lower absolute average mean curvature and shallower sulcal depth in AD subjects, where the group difference becomes more significant if we measure the quantities along the sulcal lines rather than over the entire sulcal area. The most remarkable difference in the AD patients was the average sulcal depth (control: 11.70 and AD: 11.34). PMID- 19683581 TI - Magnetization transfer phenomenon in the human brain at 7 T. AB - Magnetization transfer is an important source of contrast in magnetic resonance imaging which is sensitive to the concentration of macromolecules and other solutes present in the tissue. Magnetization transfer effects can be visualized in magnetization transfer ratio images or quantified via the z-spectrum. This paper presents methods of measuring the z-spectrum and of producing high resolution MTR images and maps of z-spectrum asymmetry in vivo at 7 T, within SAR limits. It also uses a 3-compartment model to measure chemical exchange and magnetization transfer parameters from the z-spectrum data. The peak in the z spectrum associated with chemical exchange between amide and water protons (amide proton transfer, APT, effects) is much more apparent at 7 T than at 3 T. Furthermore at 7 T quantitative APT results varied between the corpus callosum and other white matter structures, suggesting that quantitative APT imaging could be used as a method of measuring myelination. The results also suggest that chemical exchange is not responsible for the phase shift observed in susceptibility weighted images between grey matter and white matter. PMID- 19683582 TI - Alterations in the processing of non-drug-related affective stimuli in abstinent heroin addicts. AB - Long-term exposure to drug may alter the neural system associated with affective processing, as evidenced by both clinical observations and behavioral data documenting dysfunctions in emotional experiences and processing in drug addicts. Although many imaging studies examined neural responses to drug or drug-related cues in addicts, there have been few studies explicitly designed to reveal their neural abnormalities in processing non-drug-related natural affective materials. The present study asked abstinent heroin addicts and normal controls to passively view standardized affective pictures of positive, negative, or neutral valence and compared their brain activities with functional MRI. Compared to normal controls, addicts showed reduced activation in right amygdala in response to the affective pictures, consistent with previous reports of blunted subjective experience for affective stimuli in addicts. Furthermore, in two visual cortical areas BA 19 and 37, while the controls showed greater responses to positive pictures than to negative ones replicating literature findings, the addicts showed the opposite pattern. The results reveal a complex pattern of altered processing of non-drug-related affective materials in addicts showing both heightened and blunted neural responses in different brain regions and for different stimulus valence. The present study highlights the importance of brain imaging research on drug addicts' processing of affective stimuli in understanding disruptions in their emotion circuitry. PMID- 19683583 TI - Abdominal visceral fat accumulation is associated with hippocampus volume in non dementia patients with type 2 diabetes mellitus. AB - Obesity is associated with cognitive dysfunction, for which changes in the hippocampus plausibly play a pivotal role. We tested the hypothesis that an elevated level of visceral fat accumulation (VFA) correlates with hippocampus volume and insulin resistance in non-dementia patients with type 2 diabetes. Subjects included 48 non-dementia patients with type 2 diabetes, who were divided into two groups, high VFA group (mean+/-standard deviation: age=65+/-6 years, n=30) and normal VFA group (65+/-5 years, n=18). Hippocampus volume has been quantitated with computer-assisted analysis using a magnetic resonance imaging (MRI) voxel-based specific regional analysis system developed for the study of Alzheimer's disease (VSRAD), which yields a Z-score as the end point for assessment of hippocampal volume. The Z-score was higher in the high VFA group than in the normal VFA group (p<0.0001). The fasting plasma glucose (p<0.05) and insulin concentrations (p<0.0001) and the homeostasis model assessment (HOMA) index (p<0.0001) were higher in the high VFA group than in the normal VFA group. Multiple regression analysis showed that VFA levels were independently predicted by Z-score and HOMA index. Our results indicate that the elevated level of VFA in Japanese non-dementia patients with type 2 diabetes is characterized by increased hippocampus volume and insulin resistance, and that the Z-score and HOMA index are independent predictors of VFA. PMID- 19683584 TI - Investigating the predictive value of whole-brain structural MR scans in autism: a pattern classification approach. AB - Autistic spectrum disorder (ASD) is accompanied by subtle and spatially distributed differences in brain anatomy that are difficult to detect using conventional mass-univariate methods (e.g., VBM). These require correction for multiple comparisons and hence need relatively large samples to attain sufficient statistical power. Reports of neuroanatomical differences from relatively small studies are thus highly variable. Also, VBM does not provide predictive value, limiting its diagnostic value. Here, we examined neuroanatomical networks implicated in ASD using a whole-brain classification approach employing a support vector machine (SVM) and investigated the predictive value of structural MRI scans in adults with ASD. Subsequently, results were compared between SVM and VBM. We included 44 male adults; 22 diagnosed with ASD using "gold-standard" research interviews and 22 healthy matched controls. SVM identified spatially distributed networks discriminating between ASD and controls. These included the limbic, frontal-striatal, fronto-temporal, fronto-parietal and cerebellar systems. SVM applied to gray matter scans correctly classified ASD individuals at a specificity of 86.0% and a sensitivity of 88.0%. Cases (68.0%) were correctly classified using white matter anatomy. The distance from the separating hyperplane (i.e., the test margin) was significantly related to current symptom severity. In contrast, VBM revealed few significant between-group differences at conventional levels of statistical stringency. We therefore suggest that SVM can detect subtle and spatially distributed differences in brain networks between adults with ASD and controls. Also, these differences provide significant predictive power for group membership, which is related to symptom severity. PMID- 19683585 TI - Neuroticism modulates amygdala-prefrontal connectivity in response to negative emotional facial expressions. AB - Neuroticism is associated with the experience of negative affect and the development of affective disorders. While evidence exists for a modulatory role of neuroticism on task induced brain activity, it is unknown how neuroticism affects brain connectivity, especially the crucial coupling between the amygdala and the prefrontal cortex. Here we investigate this relation between functional connectivity and personality in response to negative facial expressions. Sixty healthy control participants, from the Netherlands Study on Depression and Anxiety (NESDA), were scanned during an emotional faces gender decision task. Activity and functional amygdala connectivity (psycho-physiological interaction [PPI]) related to faces of negative emotional valence (angry, fearful and sad) was compared to neutral facial expressions, while neuroticism scores were entered as a regressor. Activity for fearful compared to neutral faces in the dorsomedial prefrontal (dmPFC) cortex was positively correlated with neuroticism scores. PPI analyses revealed that right amygdala-dmPFC connectivity for angry and fearful compared to neutral faces was positively correlated with neuroticism scores. In contrast, left amygdala-anterior cingulate cortex (ACC) connectivity for angry, fearful and sad compared to neutral faces was negatively related to neuroticism levels. DmPFC activity has frequently been associated with self-referential processing in social cognitive tasks. Our results therefore suggest that high neurotic participants display stronger self-referential processing in response to negative emotional faces. Second, in line with previous reports on ACC function, the negative correlation between amygdala-ACC connectivity and neuroticism scores might indicate that those high in neuroticism display diminished control function of the ACC over the amygdala. These connectivity patterns might be associated with vulnerability to developing affective disorders such as depression and anxiety. PMID- 19683586 TI - Cerebellum development during childhood and adolescence: a longitudinal morphometric MRI study. AB - In addition to its well-established role in balance, coordination, and other motor skills, the cerebellum is increasingly recognized as a prominent contributor to a wide array of cognitive and emotional functions. Many of these capacities undergo dramatic changes during childhood and adolescence. However, accurate characterization of co-occurring anatomical changes has been hindered by lack of longitudinal data and methodologic challenges in quantifying subdivisions of the cerebellum. In this study we apply an innovative image analysis technique to quantify total cerebellar volume and 11 subdivisions (i.e. anterior, superior posterior, and inferior posterior lobes, corpus medullare, and three vermal regions) from anatomic brain MRI scans from 25 healthy females and 25 healthy males aged 5-24 years, each of whom was scanned at least three times at approximately 2-year intervals. Total cerebellum volume followed an inverted U shaped developmental trajectory peaking at age 11.8 years in females and 15.6 years in males. Cerebellar volume was 10% to 13% larger in males depending on the age of comparison and the sexual dimorphism remained significant after covarying for total brain volume. Subdivisions of the cerebellum had distinctive developmental trajectories with more phylogenetically recent regions maturing particularly late. The cerebellum's unique protracted developmental trajectories, sexual dimorphism, preferential vulnerability to environmental influences, and frequent implication in childhood onset disorders such as autism and ADHD make it a prime target for pediatric neuroimaging investigations. PMID- 19683587 TI - Phylogeny and biogeography of the rice tribe (Oryzeae): evidence from combined analysis of 20 chloroplast fragments. AB - Based on sequences of 20 chloroplast fragments, we generated a fully resolved phylogeny of Oryzeae and estimated divergence times of its major lineages as well as explored the historical biogeography of the tribe. Our results (1) confirmed the monophyly of Oryzeae and two-subtribe subdivision; (2) indicated that Maltebrunia, Potamophila and Prosphytochloa were genetically distinct enough to deserve generic status but Maltebrunia and Prosphytochloa were sister groups in the subtribe Oryzinae while Potamophila was a member in the subtribe Zizaniinae; (3) suggested that the previously unresolved phylogeny of the subtribe Zizaniinae was most likely explained by insufficient data due to rapid radiation; (4) provided the first well-corroborated timescale for the origin and divergence of Oryzeae, with the crown node of Oryzeae and the deepest split of Oryza at approximately 24 and 15MYA, respectively; (5) developed a biogeographic history of the tribe and substantiated the important roles of long-distance dispersal in the origin and diversification of the tribe Oryzeae. PMID- 19683588 TI - Evidence of chloroplast capture in South American Nothofagus (subgenus Nothofagus, Nothofagaceae). AB - Subgenus Nothofagus, although geographically restricted at present to temperate areas of South America, has captured much attention in discussions of plant biogeography due to its widespread distribution through Gondwanan continents during the Tertiary. However, phylogenetic relationships within the subgenus Nothofagus have not yet been resolved. We examined geographic patterns of intraspecific and interspecific genetic variation to detect whether incongruences in nuclear or plastid DNA phylogenies occur, in order to better understand the evolutionary history of the subgenus Nothofagus. We conducted spatially-explicit sampling at 10 distinct locations throughout the range of austral South American forests and sampled all present Nothofagus species. We used ITS and chloroplast DNA sequences to estimate phylogenetic relationships. A phylogeny constructed from nuclear genes resolved the subgenus Nothofagus as monophyletic. We found that N. antarctica was a sister to a clade of evergreen species (N. betuloides, N. dombeyi, and N. nitida), while N. pumilio likely diverged earlier. Nine cpDNA haplotypes were distinguished in the subgenus Nothofagus which were associated to geographic locations rather than to taxonomic relationships. This species independent cpDNA phylogeographic structures within the subgenus Nothofagus may be related to repeated chloroplast capture events over geological time in Patagonia. PMID- 19683589 TI - Use of 2-D electrophoresis and ESI mass spectrometry techniques to characterize Fusobacterium nucleatum proteins up-regulated after oxidative stress. AB - The genus Fusobacterium belongs to the Fusobacteriaceae family and is a Gram negative obligate anaerobic bacterium found in the human oral microbiota. Even that Fusobacterium nucleatum cannot grow under aerobic conditions, they may exhibit aerotolerance as an adaptive response which could figure as an important virulence factor, during the stages of infection, when these anaerobes are shifted to aerobic conditions. In this regard, little is known about bacterial oxidative stress adaptive response and the influence of this adaptation on the host-bacteria relationship. We aimed to use both techniques 2-DE and Electrospray Ionization Mass Spectrometry (ESI-MS) to characterize proteins in F. nucleatum, after oxidative stress. We related three different proteins which were up regulated by oxidative stress. As its genome is already sequenced, these proteins were found in data base search, by homology. Thus, by using techniques as ESI Q/TOF-MS, in addition to 2-DE, the opportunity exists to gain a more holistic view of the bacterial proteome of human pathogens, to achieve a better understanding of species diversity and to elucidate the role of specific proteins in disease. This work represents one of the first studies using genetic and physiological approaches to understand the phenomenon of oxidative stress in F. nucleatum. PMID- 19683590 TI - Aclidinium bromide provides long-acting bronchodilation in patients with COPD. AB - Aclidinium bromide is a novel, long-acting, muscarinic antagonist in phase III development for the maintenance treatment of COPD. This phase IIb study investigated the efficacy and safety of aclidinium for the treatment of moderate to severe COPD to establish the optimal dose for phase III studies. A total of 464 patients with moderate to severe stable COPD were randomised to double-blind, once-daily treatment with aclidinium (25, 50, 100, 200, or 400microg), placebo, or open-label tiotropium (18microg) for 4 weeks. Spirometric measurements were performed at 22-24h after the first dose and then at weekly intervals, and from 0.5 to 6h post-dose on day 1 and day 29. Compared with placebo, aclidinium 200microg and 400microg significantly increased trough FEV(1) on day 29 versus baseline. During the first 6h post-dose, the bronchodilatory effect of aclidinium (all doses) on day 1 was comparable to that on day 29. Time to peak FEV(1) was 3h for aclidinium 100-400microg. Aclidinium was well tolerated, with no dose dependent effect on ECG, laboratory parameters, or adverse events. The incidence of AEs was generally comparable to placebo. Aclidinium produced sustained bronchodilation over 24h and was well tolerated during this short-term study. Based on these data, aclidinium 200microg was selected as the investigational dose for future clinical trials in COPD. PMID- 19683591 TI - Additional effects of pranlukast in salmeterol/fluticasone combination therapy for the asthmatic distal airway in a randomized crossover study. AB - BACKGROUND: Salmeterol/fluticasone combination (SFC) therapy is used to control inflammation in the distal airway of patients with well-controlled asthma, but the efficacy of this approach is unclear. OBJECTIVES: The goal of the study was to evaluate the effect of pranlukast, a leukotriene receptor antagonist (LTRA), on distal airway inflammation and pulmonary resistance in patients with asthma that was well-controlled using SFC therapy alone. METHODS: The subjects were 32 patients with well-controlled asthma (age 61.1+/-17.8 years old, Step 3 in the GINA guidelines, Asthma Control Test score 23.2+/-1.8 points) based on use of SFC therapy alone for more than 3 months. These subjects were randomly assigned to groups receiving SFC alone or SFC+LTRA (pranlukast 450 mg daily) and then switched to the opposite group after 4 weeks in a crossover manner. Eosinophilic inflammation in induced sputum samples was assessed after each treatment period. Sputum was induced by inhalation of 10% hypertonic saline for 15 min. Impulse oscillometry parameters (R5, R20, X5 and AX) and spirometry were examined during each period. The Asthma-related Quality of Life Questionnaire (AQLQ) was also administered in each period. RESULTS: The ECP levels in late-phase sputum were significantly higher than those in early-phase sputum with SFC therapy alone (178.3+/-166.0 vs. 65.5+/-68.9 microg/l, p<0.001), whereas these values did not differ significantly with SFC+LTRA treatment (70.9+/-95.1 vs. 54.6+/-65.7, p=0.554). ECP levels in late-phase sputum with SFC therapy were also significantly higher than those with SFC+LTRA (p=0.045). The values of R5, R20, R5-R20 (kPa/(L/s)), and AX (kPa/L) all significantly improved during with SFC+LTRA treatment compared with SFC alone (median (25-75 percentile)): 0.350 (0.283-0.440) vs. 0.340 (0.280-0.378), p=0.036; 0.280 (0.233-0.365) vs. 0.270 (0.240-0.318), p=0.019; 0.050 (0.030-0.110) vs. 0.500 (0.030-0.073), p=0.032; and 0.570 (0.308-1.045) vs. 0.410 (0.263-0.820), p=0.014; respectively. Pulmonary function indexes did not differ significantly between the two treatments, but the symptom and activity limitation domains of the AQLQ were significantly improved by SFC+LTRA treatment. CONCLUSION: This study suggests that the combination of SFC and LTRA may give better control of residual eosinophilic inflammation in the distal airway compared with SFC therapy alone. PMID- 19683592 TI - The effects of physical exercise on the cigarette smoke-induced pulmonary oxidative response. AB - Studies have shown that the oxidative power of cigarettes is related to the pathogenesis of several pulmonary diseases and that regular physical exercise contributes significantly to reducing the deleterious effects of cigarettes. The objective of the present study was to investigate the therapeutic effects of physical exercise on histological and oxidative stress markers in animals exposed to cigarette smoke. Thirty-six male, eight-week-old C57BL-6 mice were divided into four groups (n = 9 for each group): control, exercise, cigarette smoke, and cigarette smoke plus exercise. The cigarette smoke (CS) groups were exposed to cigarette smoke 3 times/day (4 cigarettes/session) for 60 consecutive days. The exercise groups were submitted to swimming physical training 5 days/week for eight weeks. Forty-eight hours after the last exercise and cigarette exposure, the animals were sacrificed using cervical traction. The right lung was removed, processed, and stored for future analysis. In addition to the analysis of collagen content (hydroxyproline), oxidant production (anion superoxide), antioxidant enzyme activity (SOD and CAT), and lipid and protein oxidative damage (TBARS and Carbonylation), histological and morphological studies were performed. The results revealed that the animals exposed to cigarette smoke showed enlargement and destruction of the alveolar septum and increases in the numbers of macrophages and neutrophils, as well as in the amount of collagen. Our results also showed a decrease in the volume density of elastic fibers and an increase in the volume density of airspaces. However, physical exercise partially improved these markers. Additionally, physical exercise decreased oxidant production and increased the activity of the enzymatic antioxidant defense system, but did not reverse lipid and protein oxidative damage induced by cigarette smoke. These results suggest that physical training partially improves histological and oxidative stress parameters in the lungs of animals chronically exposed to cigarette smoke and that other therapies can contribute to potentiate these effects. PMID- 19683593 TI - Impact of perturbed pyruvate metabolism on adipocyte triglyceride accumulation. AB - This study aimed to test the hypothesis that adipocyte TG accumulation could be altered by specifically perturbing pyruvate metabolism. We treated cultured 3T3 L1 adipocytes with chemical inhibitors of lactate dehydrogenase (LDH) and pyruvate carboxylase (PC), and characterized their global effects on intermediary metabolism using metabolic flux and isotopomer analysis. Inhibiting the enzymes over several days did not alter the adipocyte differentiation program as assessed by the expression levels of peroxisome proliferator-activated receptor-gamma and glycerol-3-phosphate dehydrogenase. The main metabolic effects were to up regulate intracellular lipolysis and decrease TG accumulation. Inhibiting PC also up-regulated glycolysis. Flux estimates indicated that the reduction in TG was due to decreased de novo fatty acid synthesis. Exogenous addition of free fatty acids dose-dependently increased the cellular TG level in the inhibitor-treated adipocytes, but not in untreated control cells. The results of this study support our hypothesis regarding the critical role of pyruvate reactions in TG synthesis. PMID- 19683594 TI - In vitro synthesis of arachidonoyl amino acids by cytochrome c. AB - Arachidonoyl amino acids are a class of endogenous lipid messengers that are expressed in the mammalian central nervous system and peripherally. While several of their prominent pharmacologic effects have been documented, the mechanism by which arachidonoyl amino acids are biosynthesized has not been defined. We have previously observed that the mitochondrial protein, cytochrome c, is capable of catalyzing the formation of the prototypic arachidonoyl amino acid, arachidonoyl glycine, utilizing arachidonoyl CoA and glycine as substrates, in the presence of hydrogen peroxide. Here we report that cytochrome c is similarly able to catalyze the formation of N-arachidonoyl serine, N-arachidonoyl alanine, and N arachidonoyl gamma aminobutyric acid from arachidonoyl CoA and the respective amino acids. The identities of the arachidonoyl amino acid products were verified by mass spectral fragmentation pattern analysis. The synthetic reactions exhibited Michaelis-Menten kinetics and continued favorably at physiologic temperature and pH. Spectral data indicate that both cytochrome c protein structure and a +3 heme iron oxidation state are required for the reaction mechanism to proceed optimally. Reactions designed to catalyze the formation of N arachidonoyl dopamine were not efficient due to the rapid oxidation of dopamine substrate by hydrogen peroxide, consuming both reactants. Finally, under standard assay conditions, arachidonoyl CoA and ethanolamine were found to react spontaneously to form anandamide, independent of cytochrome c and hydrogen peroxide. Accordingly, it was not possible to demonstrate a potential role for cytochrome c in the biosynthetic mechanism for either arachidonoyl dopamine or anandamide. However, the ability of cytochrome c to effectively catalyze the formation of N-arachidonoyl serine, N-arachidonoyl alanine, and N-arachidonoyl gamma aminobutyric acid in vitro highlights its potential role for the generation of these lipid messengers in vivo. PMID- 19683596 TI - Canine pulmonary angiostrongylosis: the influence of climate on parasite distribution. AB - The geographic range of Angiostrongylus vasorum is expanding, leading to increased disease. Although observed cases of canine pulmonary angiostrongylosis have been dutifully reported in the literature, the state of biological knowledge remains too poor to predict future patterns of spread with any confidence. Nevertheless, there is an urgent need to identify areas that are likely to be suitable for parasite establishment. Preliminary attempts to do this using a climatic envelope approach suggest that several new areas are open to colonisation, even without invoking climate change. The risk of parasite importation into these areas should be mitigated, e.g. by restricting movement of dogs unless tested or treated for A. vasorum, and monitored by focused surveillance of definitive and intermediate hosts. These efforts will benefit from newly developed diagnostic tests. PMID- 19683597 TI - In vivo diagnosis and classification of colorectal neoplasia by chromoendoscopy guided confocal laser endomicroscopy. AB - BACKGROUND & AIMS: Colorectal cancer surveillance guidelines rely on clinicohistologic features of adenomas. Unfortunately, in common practice, recording of these features lacks precision and uniformity, which might hamper appropriate follow-up decisions. Confocal laser endomicroscopy (CLE) is a novel technology that allows real-time in vivo microscopy of the mucosa and provides accurate histopathology. The aims of this study were (1) to define and validate differential features of adenomatous and nonadenomatous colorectal polyps by chromoendoscopy-guided CLE (C-CLE) and (2) to assess predictive value of this technique for diagnosis of colorectal neoplasia. METHODS: Patients at risk for colorectal cancer were prospectively investigated by using CLE. During extubation, fluorescein 10% was used in conjunction with acriflavine hydrochloride 0.05% to characterize global tissue architecture as well as cytonuclear features of colorectal epithelium. Ex vivo histology was used as gold standard. Reproducibility tests were performed. RESULTS: In total, 116 colorectal polyps from 72 patients were examined. Ex vivo histology showed 68 adenomas, 6 invasive carcinomas, 30 hyperplastic polyps, and 12 inflammatory polyps. C-CLE of adenomas revealed lack of epithelial surface maturation, crypt budding, altered vascular pattern, and loss of cell polarity. In contrast, C-CLE of nonadenomatous polyps revealed epithelial surface maturation, and minor abnormalities of crypt architecture and of vascular pattern, and maintained cell polarity. Adenoma dysplasia score reliably discriminated high-grade dysplasia from low-grade dysplasia (accuracy, 96.7%). Interobserver agreement was high (K coefficients: pathologist, 0.92; endomicroscopist, 0.88). In vivo histology predicted ex vivo data with sensitivity of 97.3%, specificity of 92.8%, and accuracy of 95.7%. CONCLUSIONS: C-CLE accurately discriminates adenomatous from nonadenomatous colorectal polyps and enables evaluation of degree of dysplasia during ongoing endoscopy. This technology might offer considerable potential to ultimately fine tune surveillance programs, particularly in high-risk groups. PMID- 19683598 TI - Fluorescence studies on the interaction of choline-binding domain B of the major bovine seminal plasma protein, PDC-109 with phospholipid membranes. AB - The microenvironment and accessibility of the tryptophan residues in domain B of PDC-109 (PDC-109/B) in the native state and upon ligand binding have been investigated by fluorescence quenching, time-resolved fluorescence and red-edge excitation shift (REES) studies. The increase in the intrinsic fluorescence emission intensity of PDC-109/B upon binding to lysophosphatidylcholine (Lyso-PC) micelles and dimyristoylphosphatidylcholine (DMPC) membranes was considerably less as compared to that observed with the whole PDC-109 protein. The degree of quenching achieved by different quenchers with PDC-109/B bound to Lyso-PC and DMPC membranes was significantly higher as compared to the full PDC-109 protein, indicating that membrane binding afforded considerably lesser protection to the tryptophan residues of domain B as compared to those in the full PDC-109 protein. Finally, changes in red-edge excitation shift (REES) seen with PDC-109/B upon binding to DMPC membranes and Lyso-PC micelles were smaller that the corresponding changes in the REES values observed for the full PDC-109. These results, taken together suggest that intact PDC-109 penetrates deeper into the hydrophobic parts of the membrane as compared to domain B alone, which could be the reason for the inability of PDC-109/B to induce cholesterol efflux, despite its ability to recognize choline phospholipids at the membrane surface. PMID- 19683599 TI - Domain-specific determinants of catalysis/substrate binding and the oligomerization status of barley UDP-glucose pyrophosphorylase. AB - UDP-glucose (UDPG) pyrophosphorylase (UGPase) produces UDPG for sucrose and polysaccharide synthesis and glycosylation reactions. In this study, several barley UGPase mutants were produced, either single amino acid mutants or involving deletions of N- and C-terminal domains (Ncut and Ccut mutants, respectively) and of active site region ("NB loop"). The Del-NB mutant yielded no activity, whereas Ncut deletions and most of Ccut mutants, including short deletions at the so called "I-loop" region of C-terminal domain, as well as a single K260A mutant resulted in very low activity. For wt and the mutants, kinetics with UDPG were linear on reciprocal plots, whereas PPi at concentrations above 1 mM exerted strong substrate inhibition. Both K260A and most of the Ccut mutants had very high Km with PPi (up to 33 mM), whereas Ncut deletions had greatly increased Km with UDPG (up to 57 mM). Surprisingly, an 8 amino acid deletion from end of the C-terminus resulted in an enzyme (Ccut-8 mutant) with 44% higher activity when compared to wt, but with similar Km values. Whereas Ccut 8 existed solely as a monomer, other deletion mutants had a more oligomerized status, e.g. Ncut mutants existing primarily as dimers. Overall, the data confirmed the essential role of NB loop in catalysis, but also pointed out to the role of both N- and C-termini for activity, substrate binding and oligomerization. The importance of oligomerization status for enzymatic activity of UGPase is discussed. PMID- 19683600 TI - First-principles studies on alloying and simplified thermodynamic aqueous chemical stability of calcium-, zinc-, aluminum-, yttrium- and iron-doped magnesium alloys. AB - In the present study, the density functional theory implemented in the Vienna Ab initio Simulation Package has been used to investigate the alloying effects of different elements of calcium, zinc, yttrium, aluminum and iron when introduced in the Mg crystal lattice. In particular, studies have been conducted to investigate the thermodynamics of the overall hydrolysis reaction of the different alloys with pure water. Phase stability results obtained from the first principles calculations of the alloys considered are in good agreement with the published phase diagrams. The heats of the aqueous chemical reactions calculated in this study have been compared with that corresponding to the reaction of pure Mg with water. The heats of reactions dependence on the chemical compositions of the alloys have been investigated and, specifically, the role of Ca, Zn, Y, Al and Fe on the aqueous chemical stability and reactivity of these Mg alloys have been discussed. Results of these studies will help understand the biodegradable characteristic of Mg based alloys. PMID- 19683601 TI - Biphasic calcium phosphate to repair nasal septum: the first in vitro and in vivo study. AB - Our objective was to evaluate the cytocompatibility and biocompatibility of biphasic calcium phosphate (BCP) in the nasal respiratory airway. In vitro, the attachment rate was quantified on BCP disks with normal human epithelial cells at 1, 3 and 24 h by determining N-acetyl beta-D-hexosaminidase activity. Proliferative activity of cells was indirectly assessed by MTT assay at 3, 9, 15 and 21 days. Plastic surfaces were used as positive control. In vivo, 15 rabbits underwent anterior nasal septum perforation and 10 septa were repaired with BCP disks. Five non-implanted animals were sacrificed at 3 months. Two groups of five implanted animals were sacrificed at 1 and 2 months. The surface of new airway mucosa covering BCP disks was evaluated macroscopically. During both steps, light microscopy, immunohistochemistry and scanning electron microscopy were performed. Statistical analysis was performed with the Mann-Whitney U-test. In vitro, at 1 and 3 h, the attachment rates were significantly better than on the plastic surface (p < 10(-2)). Mitochondrial activity increased on both surfaces but began 6 days later than on plastic. After 21 days of culture, cells were confluent and formed a monolayer covering the implant even in the bottom of the pores. In vivo, no perforations in the control group closed spontaneously. The mean rate of closure was 63% in the 1 month group and 64% in the 2 month group (p > 0.05). Implants were invaded by inflammatory reaction covered by incomplete differentiated respiratory epithelium. Throughout the study, all immunohistochemical findings remained positive. These data suggest a good affinity between BCP and nasal epithelial cells. BCP could be used to rebuild nasal septa. PMID- 19683602 TI - Stereolithography of spatially controlled multi-material bioactive poly(ethylene glycol) scaffolds. AB - Challenges remain in tissue engineering to control the spatial, mechanical, temporal and biochemical architectures of scaffolds. Unique capabilities of stereolithography (SL) for fabricating multi-material spatially controlled bioactive scaffolds were explored in this work. To accomplish multi-material builds, a mini-vat setup was designed allowing for self-aligning X-Y registration during fabrication. The mini-vat setup allowed the part to be easily removed and rinsed, and different photocrosslinkable solutions to be easily removed and added to the vat. Two photocrosslinkable hydrogel biopolymers, poly(ethylene glycol) dimethacrylate (PEG-dma, MW 1000) and poly(ethylene glycol) diacrylate (PEG-da, MW 3400), were used as the primary scaffold materials. Multi-material scaffolds were fabricated by including controlled concentrations of fluorescently labeled dextran, fluorescently labeled bioactive PEG or bioactive PEG in different regions of the scaffold. The presence of the fluorescent component in specific regions of the scaffold was analyzed with fluorescent microscopy, while human dermal fibroblast cells were seeded on top of the fabricated scaffolds with selective bioactivity and phase contrast microscopy images were used to show specific localization of cells in the regions patterned with bioactive PEG. Multi material spatial control was successfully demonstrated in features down to 500 microm. In addition, the equilibrium swelling behavior of the two biopolymers after SL fabrication was determined and used to design constructs with the specified dimensions at the swollen state. The use of multi-material SL and the relative ease of conjugating different bioactive ligands or growth factors to PEG allows for the fabrication of tailored three-dimensional constructs with specified spatially controlled bioactivity. PMID- 19683603 TI - Premature degradation of poly(alpha-hydroxyesters) during thermal processing of Bioglass-containing composites. AB - Bioactive, biodegradable composites are increasingly being explored as bone replacement materials and as scaffolds for tissue engineering. Their properties are not only dependent on the properties of the filler and matrix, but are also determined by their interaction. This study investigated the effect on poly(D,L lactide) (PDLLA) matrix when processed at high-temperatures in the presence of Bioglass particulate filler. Composites with different filler contents were compounded at elevated temperatures by co-extrusion followed by compression moulding and compared with composites of similar composition prepared by thermally induced phase separation (TIPS), a low-temperature processing route. It was found that the inclusion of Bioglass in PDLLA under elevated temperatures resulted in the degradation of the matrix, leading to a reduction in the mechanical properties of the composites and in the molecular weight of the matrix. Attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy showed the presence of a peak at 1600 cm(-1) in the composite material, particularly when processed at elevated temperatures, whereas no peak at this wavelength was discernible for the pure PDLLA. Furthermore, time-based ATR-FTIR spectra taken at elevated temperatures on the TIPS-processed composites showed an increase in the intensity of the peak at 1600 cm(-1) and a concomitant reduction of the CO stretch peak at 1745 cm(-1) with time. This suggested the formation of a carboxylate salt by-products as a consequence of a reaction at the interface between the Bioglass filler and the PDLLA matrix. Therefore, the results confirmed that this degradation was not solely due to shear effects during the extrusion process. This work thereby supports the assertion that, in the presence of Bioglass filler particles, poly(alpha-hydroxyester)-based composites should not be processed at elevated temperatures. PMID- 19683604 TI - Effect of fluoride from glass ionomer on discoloration and corrosion of titanium. AB - The objective of the study was to examine the effects that fluoride ions released from different dental glass ionomer cements may have on titanium. The study included determination of the amounts of released ions and measurement of the color changes on titanium plates cemented with four kinds of commercial glass ionomer cements due to immersion of such "sandwich specimens" in 1% saline (NaCl) solution. The discoloration of titanium in the presence of glass ionomer cements was observed. In addition, for specimens cemented with two of the cements titanium ions were found in the solution after immersion. The results of the present study show that a low concentration of released fluoride ions and other elements from glass ionomer cements may cause aesthetic problems of discoloration of titanium restorations and appliances. PMID- 19683605 TI - Poly(lactic-co-glycolic acid) electrospun fibrous meshes for the controlled release of retinoic acid. AB - Poly(lactic-co-glycolic acid) (PLGA) meshes loaded with retinoic acid (RA) were prepared by applying the electrospinning technique. The purpose of the present work was to combine the biological effects of RA and the advantages of electrospun meshes to enhancing the mass transfer features of controlled release systems and cell interaction with polymeric scaffolds. The processing conditions for the fabrication of three-dimensional meshes were optimized by studying their influence on mesh morphology. Tensile testing showed that RA loading influenced the meshes' mechanical properties by increasing their strength and rigidity. Moreover, the drug release and degradation profiles of the electrospun systems were compared to analogous RA-loaded PLGA films prepared by solvent casting. The results of this study highlight that the electrospun meshes preserved their fibrous structure after 4 months under in vitro physiological conditions and showed a sustained controlled release of the loaded agent in comparison to that observed for cast films. The bioactivity of the loaded RA was investigated on murine preosteoblasts cells by evaluating its influence on cell proliferation and morphology. PMID- 19683606 TI - Air embolism in gastroscopy. AB - Endoscopy of the upper gastrointestinal tract (GIT) is a common medical examination. One of the rare but serious, albeit fatal complications of gastroscopy is venous air embolism. We performed a literature search with the keywords "air embolism", "gastroscopy", and "endoscopy". There were 14 cases of air embolism associated with gastroscopy. The median age was 66 years old (range 4 months-80 years old). The main presenting symptoms were neurological (n=9) and respiratory compromise (n=7). The main investigation used for diagnosis were CT (n=10) and ECHO (n=6). The main risk factor identified was mucosal breach (n=9). Hyperbaric oxygen therapy was used in four cases. The mortality rate is 57.1%. Air embolism is a very rare complication and is often overlooked. Rapid diagnosis is vital for successful treatment. It should be considered in any patient with sudden onset of severe cardiopulmonary and/or neurologic decompensation during gastroscopy. PMID- 19683607 TI - Are we performing enough emergency laparoscopic cholecystectomies? An experience from a district general hospital. PMID- 19683608 TI - Development of criteria for a diagnosis: lessons from the night eating syndrome. AB - Criteria for inclusion of diagnoses of Axis I disorders in the forthcoming Diagnostic and Statistical Manual (DSM-V) of the American Psychiatric Association are being considered. The 5 criteria that were proposed by Blashfield et al as necessary for inclusion in DSM-IV are reviewed and are met by the night eating syndrome (NES). Seventy-seven publications in refereed journals in the last decade indicate growing recognition of NES. Two core diagnostic criteria have been established: evening hyperphagia (consumption of at least 25% of daily food intake after the evening meal) and/or the presence of nocturnal awakenings with ingestions. These criteria have been validated in studies that used self-reports, structured interviews, and symptom scales. Night eating syndrome can be distinguished from binge eating disorder and sleep-related eating disorder. Four additional features attest to the usefulness of the diagnosis of NES: (1) its prevalence, (2) its association with obesity, (3) its extensive comorbidity, and (4) its biological aspects. In conclusion, research on NES supports the validity of the diagnosis and its inclusion in DSM-V. PMID- 19683609 TI - Sex-specific predictors of criminal recidivism in a representative sample of incarcerated youth. AB - OBJECTIVE: The objective of the study was to identify sex-specific psychopathologic predictors of criminal recidivism among a representative sample of incarcerated youths. METHOD: In this prospective longitudinal study, the Mini International Psychiatric Interview for children and adolescents was used to assess psychopathology in juveniles entering an Austrian pretrial detention facility between March 2003 and January 2005. From the beginning of the study until January 2006, data on criminal history were obtained from the Integrierte Vollzugsverwaltung, a database containing criminal information of every individual incarcerated in Austria. Of the 370 eligible participants, the final study sample comprised 328 juveniles (56 girls and 272 boys, age range = 14-21 years, mean = 16.7). RESULTS: Reincarceration rates within the specified follow up period were 52.6% for the boys and 37.5% for the girls. Using Cox forward stepwise regression and Kaplan-Meier analyses, age at first incarceration (B = .296, Wald statistic = 17.11, P < .001) and oppositional defiant disorder (B = .751, Wald statistic = 19.25, P < .001) were identified as significant predictors for reoffending in boys. In girls, generalized anxiety disorder (B = 1.97, Wald statistic = 13.71, P < .001) was found to be a predictor for reoffending, whereas dysthymia (B = -1.44, Wald statistic = 4.02, P = .045) was found to serve as protective factor. CONCLUSION: Our study confirms high rates of reoffending after release from correctional facilities in both sexes. It further defines sex specific psychopathologic risk factors for relapse in incarcerated juveniles. According to our results, in boys, oppositional defiant disorder and early age at first incarceration are predictive of reincarceration. In girls, anxiety disorder was found to be a risk factor for future offending, whereas dysthymia was found to have a protective influence. Consequently, rehabilitation programs should be sex specific. PMID- 19683610 TI - Quality of life across the schizotypy spectrum: findings from a large nonclinical adult sample. AB - OBJECTIVE: It is well documented that patients with schizophrenia have impoverished quality of life (QOL). Efforts to determine the underpinnings of this impoverishment have implicated negative symptoms more than positive or disorganized symptoms. However, only a minority of individuals with the liability to schizophrenia will ever show manifest illness, and it is presently unclear the degree to which QOL is affected in individuals with subclinical symptoms of the disorder (ie, schizotypy). The present study examined the relative contributions of negative, positive, and disorganized schizotypy symptoms to QOL. METHODS: Measures of schizotypal symptoms and subjective and objective QOL were obtained from a sample of 1395 adults. RESULTS: Measures of schizotypal symptoms significantly corresponded to all measures of QOL, although the magnitude of correlations were significantly larger for subjective than objective measures. The negative symptom dimension explained a substantial portion of unique variance in the social domains of QOL above and beyond that accounted for by the other schizotypy dimensions. CONCLUSIONS: These findings highlight the deleterious impact of schizotypal symptoms, particularly negative symptoms. Further research clarifying the mechanism underlying this relationship is called for. PMID- 19683611 TI - Suicidality, aggression, and other treatment considerations among pregnant, substance-dependent women with posttraumatic stress disorder. AB - Posttraumatic stress disorder (PTSD) and other Axis I comorbidity among women with substance use disorders (SUDs) appear similarly prevalent and are associated with comparable negative clinical profiles and treatment outcomes. The relative contribution of comorbid PTSD vs other Axis I psychiatric disorders to clinical characteristics is largely unexamined, however, despite theory and empirical data indicating that PTSD and SUDs may have a unique relationship that confers specific risk for clinical severity and poor treatment outcome. In a sample of pregnant, opioid- and/or cocaine-dependent women entering substance abuse treatment, women with PTSD (SUD-PTSD; n = 23) were compared to those with other Axis I comorbidity (SUD-PSY; n = 45) and those without Axis I comorbidity (SUD only; n = 37). Data were collected via face-to-face interviews and urinalysis drug assays. Although the study groups had similar substance use severity, the SUD-PTSD group was more likely to report suicidality, aggression, and psychosocial impairment than both the SUD-PSY and SUD-only groups. Findings indicate treatment considerations for substance-dependent women with PTSD are broader and more severe than those with other Axis I conditions or substance dependence alone. PMID- 19683612 TI - The German version of the Toronto Structured Interview for Alexithymia: factor structure, reliability, and concurrent validity in a psychiatric patient sample. AB - BACKGROUND: Recently, the Toronto Structured Interview for Alexithymia (TSIA) was developed to supplement the self-assessment of alexithymia and/or offer a different method of measuring the alexithymia construct. The aim of this study was to evaluate the psychometric properties of a German language translation of the TSIA in a psychiatric patient sample. METHODS: Translation and back translation were performed until a high agreement of cross-language equivalence was obtained between the German and the original English language version of the TSIA. The TSIA and the German language version of the 20-Item Toronto Alexithymia Scale were administered to 237 psychiatric patients at the departments of psychiatry and psychotherapy in Germany and Switzerland. Videotapes of some of the interviews were recorded for the assessment of interrater reliability. RESULTS: The German version of the TSIA and its 4 scales correlated significantly with the German version of the 20-Item Toronto Alexithymia Scale and its 3 factor scales, providing support for concurrent validity of the interview. Confirmatory factor analyses supported the hierarchical, 4-factor structure obtained with the original English version, with 4 lower-order factors nested within 2 higher-order latent factors. Acceptable levels of internal reliability and interrater reliability were also demonstrated. CONCLUSION: The TSIA is a valid and reliable measure for assessing alexithymia, at least in clinical samples. The TSIA, together with a self-report alexithymia scale, allow for a multimethod approach to assessing alexithymia. PMID- 19683613 TI - Perfectionism and sensory phenomena: phenotypic components of obsessive compulsive disorder. AB - BACKGROUND: The aim of the study was to investigate how perfectionism and sensory phenomena (SP) interact as possible phenotypic components of obsessive-compulsive disorder (OCD). METHODS: Forty-seven adult outpatients, meeting Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, criteria for OCD, and a control group of 41 community subjects were assessed using the Frost Multidimensional Perfectionism Scale (FMPS), the University of Sao Paulo-Sensory Phenomena Scale, and other standard measures of OCD severity. RESULTS: Three of the FMPS subscales ("concern over mistakes," "doubts about action," and "parental criticism") were significantly different between OCD patients and control subjects. All subtypes of SP were significantly more frequent and more severe in OCD than in control subjects. The "incompleteness" subtype of SP was associated with high scores on all dimensions of the FMPS, whereas the "just-right" subtype of SP was only associated with "doubts about action," "personal standards," and "organization" subscales of the FMPS. CONCLUSIONS: Presence and severity of SP and specific elements of perfectionism clearly distinguish OCD patients from healthy control subjects. Some SP subtypes are associated with specific FPMS subscale scores, whereas others are not. These results emphasize the relevance of assessing different subtypes of perfectionism and SP in OCD patients as important subcomponents of the OCD phenotype. PMID- 19683614 TI - Obsessive-compulsive symptoms and disorder in patients with schizophrenia treated with clozapine or haloperidol. AB - We conducted a cross-sectional study to compare the prevalence and severity of obsessive-compulsive symptoms (OCSs) and obsessive-compulsive disorder (OCD) in patients with schizophrenia treated with clozapine or haloperidol. Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Axis I disorders-patient edition was used to diagnose schizophrenia and OCD. Sixty subjects, 40 of them using clozapine and 20 using haloperidol, completed the Yale-Brown Obsessive-Compulsive Scale, the Positive and Negative Syndrome Scale (PANSS), and the Clinical Global Impression. The prevalence of OCD in patients taking clozapine was 20%, whereas the prevalence of patients taking haloperidol was 10%, although this difference was not statistically significant (P = .540). However, patients using clozapine showed higher severity of OCSs than patients using haloperidol (P = .027) did. When schizophrenia patients were divided according to the presence or absence of OCD or OCSs, patients with schizophrenia and OCD or OCSs showed higher severity of schizophrenia symptoms when compared to those with schizophrenia without OCD and OCSs (P = .002). A PANSS total score higher than 70 and the use of antidepressants were predictors of the presence of OCSs or OCD. Schizophrenia patients taking clozapine had higher severity scores both in obsessive-compulsive and schizophrenia rating scales. These results may support an association between the exacerbation of obsessive-compulsive phenomena and the use of clozapine. PMID- 19683615 TI - Affect recognition as an independent social function determinant in schizophrenia. AB - INTRODUCTION: Facial affect recognition deficits may represent specific deficits and contribute to social dysfunction in patients with schizophrenia. Whether their impacts on social dysfunction are independent to those caused by deficits in basic neurocognition and clinical symptoms needs to be further delineated. METHOD: Association patterns between affect recognition and basic neurocognitive abilities in 40 acute and 33 stable patients with schizophrenia were compared to explore whether their interrelationships changed across clinical stages. The independent contribution of affect recognition deficits to social dysfunction was explored by multivariate models controlling for general intellectual ability, basic neurocognition, and clinical symptoms. RESULTS: Affect recognition deficits were associated with social role performances, self-care, and contributed independently to global social functioning in stable patients but not in acute patients. Conversely, affect recognition deficits were associated with impaired basic neurocognitions in acute patients but not in stable patients. CONCLUSION: In stabilized community patients with schizophrenia, affect recognition deficits were relatively independent to basic neurocognition and had significant social functional consequences. PMID- 19683616 TI - Treatment response in psychotic patients classified according to social and clinical needs, drug side effects, and previous treatment; a method to identify functional remission. AB - BACKGROUND: Various approaches have been made over the years to classify psychotic patients according to inadequate treatment response, using terms such as treatment resistant or treatment refractory. Existing classifications have been criticized for overestimating positive symptoms; underestimating residual symptoms, negative symptoms, and side effects; or being to open for individual interpretation. The aim of this study was to present and evaluate a new method of classification according to treatment response and, thus, to identify patients in functional remission. METHOD: A naturalistic, cross-sectional study was performed using patient interviews and information from patient files. The new classification method CANSEPT, which combines the Camberwell Assessment of Need rating scale, the Udvalg for Kliniske Undersogelser side effect rating scale (SE), and the patient's previous treatment history (PT), was used to group the patients according to treatment response. CANSEPT was evaluated by comparison of expected and observed results. RESULTS: In the patient population (n = 123), the patients in functional remission, as defined by CANSEPT, had higher quality of life, fewer hospitalizations, fewer psychotic symptoms, and higher rate of workers than those with the worst treatment outcome. CONCLUSION: In the evaluation, CANSEPT showed validity in discriminating the patients of interest and was well tolerated by the patients. CANSEPT could secure inclusion of correct patients in the clinic or in research. PMID- 19683617 TI - Factor structure and diagnostic efficiency of the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, criteria for avoidant personality disorder in Hispanic men and women with substance use disorders. AB - OBJECTIVE: This study examined the internal consistency, factor structure, and diagnostic efficiency of the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV), criteria for avoidant personality disorder (AVPD) and the extent to which these metrics may be affected by sex. METHOD: Subjects were 130 monolingual Hispanic adults (90 men, 40 women) who had been admitted to a specialty clinic that provides psychiatric and substance abuse services to Spanish-speaking patients. All were reliably assessed with the Spanish-Language Version of the Diagnostic Interview for DSM-IV Personality Disorders. The AVPD diagnosis was determined by the best-estimate method. After evaluating internal consistency of the AVPD criterion set, an exploratory factor analysis was performed using principal components extraction. Afterward, diagnostic efficiency indices were calculated for all AVPD criteria. Subsequent analyses examined men and women separately. RESULTS: For the overall group, internal consistency of AVPD criteria was good. Exploratory factor analysis revealed a 1-factor solution (accounting for 70% of the variance), supporting the unidimensionality of the AVPD criterion set. The best inclusion criterion was "reluctance to take risks," whereas "interpersonally inhibited" was the best exclusion criterion and the best predictor overall. When men and women were examined separately, similar results were obtained for both internal consistency and factor structure, with slight variations noted between sexes in the patterning of diagnostic efficiency indices. CONCLUSIONS: These psychometric findings, which were similar for men and women, support the construct validity of the DSM-IV criteria for AVPD and may also have implications for the treatment of this particular clinical population. PMID- 19683618 TI - Validation of the Mood Disorder Questionnaire in the general population in Hong Kong. AB - OBJECTIVE: The objective of the study was to determine the reliability and validity of a Chinese version of the Mood Disorder Questionnaire (MDQ) in the general population in Hong Kong. METHODS: One thousand five hundred eighteen adults aged at least 18 years were randomly drawn from the general population and interviewed using a lay-administered version of the MDQ. A subsample of 114 randomly selected participants received a telephone-based Structured Clinical Interview for the Diagnostic and Statistical Manual of Mental Disorder, Fourth Edition. Six subjects were diagnosed to have bipolar disorder (bipolar I = 1, bipolar II = 4, and bipolar not otherwise specified = 1), 13 had substance/alcohol use disorder, and 95 had neither bipolar nor substance/alcohol use disorder. We determined the internal consistency, factor structure, and concurrent validity of the Chinese MDQ. RESULTS: The Cronbach alpha coefficient of the Chinese MDQ was 0.78. Principal component analysis with varimax rotation indicated a "euphoria-irritability-racing thoughts" factor, an "energized activity" factor, and a "risky behavior" factor, which explained 46.6% of the rotated variance. There were significant differences in MDQ scores between the bipolar and non-bipolar non-substance/alcohol use disorder groups. The performance of the Chinese MDQ for detecting bipolar disorder was limited by a low sensitivity. Using the original cutoff criterion, defined as clustering of 7 or more symptoms that caused moderate or severe problems, the sensitivity was 0%, whereas the overall specificity was 95.4%. Lowering the cutoff criterion to a symptom score of at least 7 that caused minor or more problems yielded the best sensitivity (0.50) and specificity (0.92). CONCLUSION: The Chinese MDQ is a reliable and valid measure of bipolar disorder in the community. PMID- 19683619 TI - Description and validation of the Affective Temperament Questionnaire. AB - BACKGROUND: Akiskal and Mallya (Psychopharmacol Bull. 1987;23:68-73) proposed criteria defining 4 affective temperaments-hyperthymic, irritable, cyclothymic, and dysthymic. This study aims to develop and validate, using a 3-point rating scale, a short questionnaire that assesses these temperaments. METHODS: The Affective Temperament Questionnaire (ATQ) was administered to a family-based sample of individuals with major depressive disorder (MDD), bipolar disorder (BP), or no mood disorder (N = 378). Factor analyses, internal consistency, and analysis of variance were undertaken to examine the factorial structure and concurrent validity (relative to Axis I mood disorder diagnosis) of the ATQ. Affective Temperament Questionnaire data were evaluated with respect to raw scores and dominant affective temperament. RESULTS: Three factors emerged- hyperthymia, cyclothymia, and dysthymia--which had moderate to high internal consistency. Support for the concurrent validity of ATQ was found, whereby temperament scores and rates of dominant affective temperaments differed with respect to mood disorder diagnosis. Hyperthymia and cyclothymia were more prevalent among individuals with BP than among individuals with MDD or no history of a mood disorder. Dysthymia occurred at a relatively similar rate among individuals with MDD or BP. CONCLUSIONS: Our findings support the use of the ATQ for collecting information regarding affective temperaments and for furthering understanding regarding the links between affective temperament and mood disorders. PMID- 19683620 TI - A multidimensional spectrum approach to post-traumatic stress disorder: comparison between the Structured Clinical Interview for Trauma and Loss Spectrum (SCI-TALS) and the Self-Report instrument (TALS-SR). AB - Dimensional approaches to psychiatric disorders have shown an increased relevance in the ongoing debate for the forthcoming Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition. In line with previously validated instruments for the assessment of different mood, anxiety, eating and psychotic spectra, we tested the validity and reliability of a newly developed Structured Clinical Interview for Trauma and Loss Spectrum (SCI-TALS). The instrument is based on a multidimensional approach to post-traumatic stress spectrum that includes a range of threatening or frightening experiences, as well as a variety of potentially significant losses, to which an individual can be exposed. Furthermore, it explores the spectrum of the peritraumatic reactions and post-traumatic symptoms that may ensue from either type of life events, targeting soft signs and subthreshold conditions, as well as temperamental and personality traits that may constitute risk factors for the development of the disorder. The aim of the present study is to describe the reliability of the self-report version of the SCI-TALS: the TALS-SR. Thirty patients with PTSD and thirty healthy control subjects were assessed with the Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition. Half of the patients and controls received the TALS-SR first and the SCI-TALS after 15 days; for the other half of the sample, the order of administration was reversed. Agreement between the self-report and the interview formats was substantial. Intraclass correlation coefficients ranged from 0.934 to 0.994, always exceeding the threshold of 0.90. Our findings provide substantial support for the reliability of the TALS-SR questionnaire. PMID- 19683623 TI - Heart disease: breaking down barriers. PMID- 19683621 TI - Re: Infrequent SMARCB1/INI1 gene alteration in epithelioid sarcoma: a useful tool in distinguishing epithelioid sarcoma from malignant rhabdoid tumor: Direct evidence of mutational inactivation of SMARCB1/INI1 in epithelioid sarcoma. PMID- 19683624 TI - Research and ethics in China. PMID- 19683625 TI - USAID: an agency without leader and direction. PMID- 19683626 TI - Time to lower treatment BP targets for hypertension? PMID- 19683627 TI - Left atrial appendage occlusion in non-valvular atrial fibrillation. PMID- 19683628 TI - Albuminuria in heart failure: a CHARMing new risk factor? PMID- 19683629 TI - Key issues for global cardiovascular medicine. PMID- 19683630 TI - Who is ineligible for warfarin in atrial fibrillation? PMID- 19683631 TI - Online social networks and wellbeing. PMID- 19683632 TI - Insulin glargine and cancer: another side to the story? PMID- 19683633 TI - Intensive glucose control and cardiovascular outcomes. PMID- 19683634 TI - Intensive glucose control and cardiovascular outcomes. PMID- 19683635 TI - Intensive glucose control and cardiovascular outcomes. PMID- 19683636 TI - Intensive glucose control and cardiovascular outcomes. PMID- 19683640 TI - Albuminuria in chronic heart failure: prevalence and prognostic importance. AB - BACKGROUND: Increased excretion of albumin in urine might be a marker of the various pathophysiological changes that arise in patients with heart failure. Therefore our aim was to assess the prevalence and prognostic value of a spot urinary albumin to creatinine ratio (UACR) in patients with heart failure. METHODS: UACR was measured at baseline and during follow-up of 2310 patients in the Candesartan in Heart failure: Assessment of Reduction in Mortality and morbidity (CHARM) Programme. The prevalence of microalbuminuria and macroalbuminuria, and the predictive value of UACR for the primary composite outcome of each CHARM study--ie, death from cardiovascular causes or admission to hospital with worsening heart failure--and death from any cause were assessed. FINDINGS: 1349 (58%) patients had a normal UACR, 704 (30%) had microalbuminuria, and 257 (11%) had macroalbuminuria. The prevalence of increased UACR was similar in patients with reduced and preserved left ventricular ejection fractions. Patients with an increased UACR were older, had more cardiovascular comorbidity, worse renal function, and a higher prevalence of diabetes mellitus than did those with normoalbuminuria. However, a high prevalence of increased UACR was still noted among patients without diabetes, hypertension, or renal dysfunction. Elevated UACR was associated with increased risk of the composite outcome and death even after adjustment for other prognostic variables including renal function, diabetes, and haemoglobin A1c. The adjusted hazard ratio (HR) for the composite outcome in patients with microalbuminuria versus normoalbuminuria was 1.43 (95% CI 1.21-1.69; p<0.0001) and for macroalbuminuria versus normoalbuminuria was 1.75 (1.39-2.20; p<0.0001). The adjusted values for death were 1.62 (1.32-1.99; p<0.0001) for microalbuminuria versus normoalbuminuria, and 1.76 (1.32-2.35; p=0.0001) for macroalbuminuria versus normoalbuminuria. Treatment with candesartan did not reduce or prevent the development of excessive excretion of urinary albumin. INTERPRETATION: Increased UACR is a powerful and independent predictor of prognosis in heart failure. FUNDING: AstraZeneca. PMID- 19683641 TI - Hypoplastic left heart syndrome. AB - Hypoplastic left heart syndrome is a rare congenital heart defect in which the left side of the heart is underdeveloped. Surgical management of hypoplastic left heart syndrome has changed the prognosis of the condition that was previously regarded as fatal. We discuss surgical strategies based on staged procedures, with the right ventricle supporting both systemic and pulmonary circulation. We also discuss other management options, such as neonatal transplantation and the recent innovation of hybrid techniques. Surgical techniques and the understanding of the pathophysiology of this condition have been at the forefront of neonatal cardiac surgery and intensive care. The management of the syndrome remains a challenge because affected children grow into adolescence and adulthood posing various new problems and demands. PMID- 19683642 TI - Antithrombotic management of patients with prosthetic heart valves: current evidence and future trends. AB - Over 4 million people worldwide have received a prosthetic heart valve, and an estimated 300,000 valves are being implanted every year. Prosthetic heart valves improve quality of life and survival of patients with severe valvular heart disease, but the need for antithrombotic therapy to prevent thrombotic complications in valve recipients poses challenges for clinicians and patients. Here, we review antithrombotic therapies for patients with prosthetic heart valves and management of thromboembolic complications. Advances in antithrombotic therapy and valve technologies are likely to improve the management of patients with prosthetic heart valves in developed countries, but the most important unmet need and potential for benefit from these new therapies is in developing countries where a massive and rapidly increasing burden of valvular heart disease exists. PMID- 19683639 TI - Percutaneous closure of the left atrial appendage versus warfarin therapy for prevention of stroke in patients with atrial fibrillation: a randomised non inferiority trial. AB - BACKGROUND: In patients with non-valvular atrial fibrillation, embolic stroke is thought to be associated with left atrial appendage (LAA) thrombi. We assessed the efficacy and safety of percutaneous closure of the LAA for prevention of stroke compared with warfarin treatment in patients with atrial fibrillation. METHODS: Adult patients with non-valvular atrial fibrillation were eligible for inclusion in this multicentre, randomised non-inferiority trial if they had at least one of the following: previous stroke or transient ischaemic attack, congestive heart failure, diabetes, hypertension, or were 75 years or older. 707 eligible patients were randomly assigned in a 2:1 ratio by computer-generated randomisation sequence to percutaneous closure of the LAA and subsequent discontinuation of warfarin (intervention; n=463) or to warfarin treatment with a target international normalised ratio between 2.0 and 3.0 (control; n=244). Efficacy was assessed by a primary composite endpoint of stroke, cardiovascular death, and systemic embolism. We selected a one-sided probability criterion of non-inferiority for the intervention of at least 97.5%, by use of a two-fold non inferiority margin. Serious adverse events that constituted the primary endpoint for safety included major bleeding, pericardial effusion, and device embolisation. Analysis was by intention to treat. This study is registered with Clinicaltrials.gov, number NCT00129545. FINDINGS: At 1065 patient-years of follow up, the primary efficacy event rate was 3.0 per 100 patient-years (95% credible interval [CrI] 1.9-4.5) in the intervention group and 4.9 per 100 patient-years (2.8-7.1) in the control group (rate ratio [RR] 0.62, 95% CrI 0.35-1.25). The probability of non-inferiority of the intervention was more than 99.9%. Primary safety events were more frequent in the intervention group than in the control group (7.4 per 100 patient-years, 95% CrI 5.5-9.7, vs 4.4 per 100 patient-years, 95% CrI 2.5-6.7; RR 1.69, 1.01-3.19). INTERPRETATION: The efficacy of percutaneous closure of the LAA with this device was non-inferior to that of warfarin therapy. Although there was a higher rate of adverse safety events in the intervention group than in the control group, events in the intervention group were mainly a result of periprocedural complications. Closure of the LAA might provide an alternative strategy to chronic warfarin therapy for stroke prophylaxis in patients with non-valvular atrial fibrillation. FUNDING: Atritech. PMID- 19683638 TI - Usual versus tight control of systolic blood pressure in non-diabetic patients with hypertension (Cardio-Sis): an open-label randomised trial. AB - BACKGROUND: The level to which systolic blood pressure should be controlled in hypertensive patients without diabetes remains unknown. We tested the hypothesis that tight control compared with usual control of systolic blood pressure would be beneficial in such patients. METHODS: In this randomised open-label trial undertaken in 44 centres in Italy, 1111 non-diabetic patients with systolic blood pressure 150 mm Hg or greater were randomly assigned to a target systolic blood pressure of less than 140 mm Hg (usual control; n=553) or less than 130 mm Hg (tight control; n=558). After stratification by centre, we used a computerised random function to allocate patients to either group. Observers who were unaware of randomisation read electrocardiograms and adjudicated events. Open-label agents were used to reach the randomised targets. The primary endpoint was the rate of electrocardiographic left ventricular hypertrophy 2 years after randomisation. Analysis was by intention to treat. This study is registered with ClinicalTrials.gov, number NCT00421863. RESULTS: Over a median follow-up of 2.0 years (IQR 1.93-2.03), systolic and diastolic blood pressure were reduced by a mean of 23.5/8.9 mm Hg (SD 10.6/7.0) in the usual-control group and by 27.3/10.4 mm Hg (11.0/7.5) in the tight-control group (between-group difference 3.8 mm Hg systolic [95% CI 2.4-5.2], p<0.0001; and 1.5 mm Hg diastolic [0.6-2.4]; p=0.041). The primary endpoint occurred in 82 of 483 patients (17.0%) in the usual-control group and in 55 of 484 patients (11.4%) of the tight-control group (odds ratio 0.63; 95% CI 0.43-0.91; p=0.013). A composite cardiovascular endpoint occurred in 52 (9.4%) patients in the usual-control group and in 27 (4.8%) in the tight control group (hazard ratio 0.50, 95% CI 0.31-0.79; p=0.003). Side-effects were rare and did not differ significantly between the two groups. INTERPRETATION: Our findings lend support to a lower blood pressure goal than is recommended at present in non-diabetic patients with hypertension. FUNDING: Boehringer Ingelheim, Sanofi-Aventis, Pfizer. PMID- 19683645 TI - Endoscopy. Preface. PMID- 19683644 TI - Beans in the pericardium. PMID- 19683643 TI - Drug-induced fibrotic valvular heart disease. AB - The initial association between the development of valvular heart disease and drugs stems from observations made during the use of methysergide and ergotamine for migraine prophylaxis in the 1960s. Since then, the appetite suppressants fenfluramine and dexfenfluramine, the dopamine agonists pergolide and cabergoline, and more recently, the recreational drug ecstasy (3,4 methylenedioxymethamphetamine; MDMA) have been implicated. Results from clinical trials show that drug dose and treatment duration affect both the risk of developing the disease and its severity. The natural history of the disease remains unclear, although regression of valvular lesions after the end of treatment has been reported. Interference with serotonin metabolism and its associated receptors and transporter gene seems a likely mechanism for development of the drug-induced valvular heart disease. Physicians need to balance the benefits of continued therapy with these drugs against possible risks. Further investigation is needed to assist with treatment decisions. Continued vigilance is necessary because several commonly prescribed treatments interact with serotonergic pathways. PMID- 19683646 TI - Equipment and instrumentation in veterinary endoscopy. AB - Endoscopic procedures are minimally invasive in nature, and have been found to decrease the postoperative stress response and postoperative pain compared with similar procedures performed by an open approach. There is an ongoing effort to make minimally invasive surgery even less invasive through research and the development of new and improved medical devices. This article provides a general overview of the necessary equipment and instrumentation that will assist practitioners in making decisions for the incorporation of endoscopy/endoscopic surgery into their practice. PMID- 19683647 TI - Anesthesia for endoscopy in small animals. AB - This article discusses considerations for general anesthesia for various endoscopic procedures in small animals. Specific drug and monitoring recommendations are made. Special physiologic concerns of individual procedures affecting the anesthetized patient are discussed. PMID- 19683648 TI - Diagnostic rigid endoscopy: otoscopy, rhinoscopy, and cystoscopy. AB - Diagnostic examinations are markedly improved by using rigid endoscopy in the ear, nasal cavity, and urinary tract. This article presents the procedure, equipment, indications, and examples of abnormalities of rigid endoscopy in these areas. Textbooks, "hands-on" courses, and in-hospital training are methods for learning about these techniques and their applications. In addition to improving diagnostics, endoscopy can also be used for therapy in these three body areas. PMID- 19683649 TI - Airway evaluation and flexible endoscopic procedures in dogs and cats: laryngoscopy, transtracheal wash, tracheobronchoscopy, and bronchoalveolar lavage. AB - Flexible endoscopy is a valuable diagnostic approach to the upper and lower respiratory tract, because it allows direct visualization and sample collection. Techniques requiring a range of specialized equipment and varying levels of experience have been developed to access and evaluate each anatomic region. Familiarity with appropriate indications for each procedure and normal appearance, cytology, and culture results from each region will enhance diagnostic success. PMID- 19683650 TI - Flexible endoscopy in small animals. AB - Flexible endoscopy is a valuable tool for the diagnosis of many small animal digestive tract diseases. This article provides a basic introduction to small animal gastrointestinal endoscopy including its diagnostic advantages as well as its limitations and complications. Although proficiency in endoscopic techniques can only be obtained through many hours of practice, this article should also encourage and stimulate the novice endoscopist. PMID- 19683651 TI - Gastrointestinal laparoscopy in small animals. AB - Since 1999, when the author first described the research and potential applications of minimally invasive gastrointestinal surgery in animals, veterinarians have begun to apply some of these techniques in treating client owned animals. Minimally invasive surgery is advocated with diagnostic, prophylactic, and therapeutic intent. There has been a transition from a minimally invasive caseload toward the expansion of diagnostic procedures, adoption of prophylactic procedures (such as lap-assisted gastropexy), and performing more difficult therapeutic procedures. Small animal patients benefit from reduced tissue trauma and experience a rapid recovery. In this article, current research and minimally invasive gastrointestinal procedures in animals are discussed. PMID- 19683652 TI - Advanced laparoscopic procedures (hepatobiliary, endocrine) in dogs and cats. AB - This article discusses several advanced laparoscopic procedures that have now been described in clinical veterinary patients. Laparoscopic-assisted cholecystostomy tube placement, laparoscopic cholecystectomy, and adrenalectomy can all be performed safely and efficiently. Case selection guidelines as well as indications, techniques, and possible complications are discussed in detail. PMID- 19683653 TI - Complications and need for conversion to laparotomy in small animals. AB - Laparoscopic procedures provide the advantage of decreased patient morbidity with improved visualization and rapid patient recovery. Complications associated with laparoscopic procedures are discussed. Conversion to open laparotomy may depend on a variety of factors related to the patient, procedure, and surgeon. There are few contraindications for performing laparoscopic procedures, but complications or conversions to an open laparotomy may be expected in a percentage of patients. PMID- 19683654 TI - Small animal exploratory thoracoscopy. AB - Exploratory thoracoscopy in small animal veterinary medicine is, compared with an open exploratory thoracotomy, a minimally invasive diagnostic procedure with several benefits. Specific indications, patient positioning, and anesthetic considerations are discussed, as well as instrumentation and general techniques for endoscopic intrathoracic exploration and biopsy. PMID- 19683655 TI - Interventional thoracoscopy in small animals. AB - Thoracoscopy is a minimally invasive technique for viewing the internal structures of the thoracic cavity. The procedure uses a rigid telescope placed through a portal and positioned in the thoracic wall to examine the contents of the pleural cavity. Once the telescope is in place, either biopsy forceps or an assortment of surgical instruments can be introduced into the thoracic cavity through adjacent portals in the thoracic wall to perform various diagnostic or surgical procedures. The minimal invasiveness of the procedure, the rapid patient recovery, and the diagnostic accuracy make thoracoscopy an ideal technique compared with other more invasive procedures. This article discusses the use of interventional thoracoscopy (an emerging surgical technique) in veterinary surgery to perform pericardial window, subtotal pericardiectomy, or lung lobectomy to correct vascular ring anomalies, to ligate patent ductus arteriosus and the thoracic duct, and to aid in the treatment of pyothorax. Most procedures are performed under thoracoscopy, and some procedures can be thoracoscopically assisted. PMID- 19683656 TI - Complications and need for conversion from thoracoscopy to thoracotomy in small animals. AB - Thoracoscopy is useful for the diagnosis and treatment of many conditions in veterinary patients. It decreases patient morbidity and improves visualization and lighting of structures within the thorax due to the magnification and lighting adjacent to the structures evaluated. The complications of thoracoscopy are described, as is the need for converting to an open thoracotomy. Complications and the need for conversion depend on the patient and the procedure performed. Procedural complications are not discussed unless they specifically relate to thoracoscopy. As confidence is gained with thoracoscopy, the need for conversion may decrease over time. However, conversions may be required more often as the degree of difficulty of thoracoscopic procedures increases. PMID- 19683657 TI - Essential criteria for evaluation of pneumococcal conjugate vaccine candidates. AB - In 2003, the World Health Organization recommended a concentration of enzyme linked immunosorbent assay (ELISA) immunoglobulin G (IgG) anti-capsular antibody of 0.35 microg/mL as a reference value that correlates to protection against invasive pneumococcal disease. This threshold can be used to demonstrate immunologic non-inferiority of a new vaccine in comparison trials that use pneumococcal conjugate vaccines (PCVs) as a comparator. An investigational 13 valent PCV (PCV13) compared with seven-valent PCV (PCV7) demonstrated that all of the pneumococcal serotypes contained in PCV13 were immunogenic, with 88-98% of vaccinees achieving antibody concentration > or = 0.35 microg/mL for serotypes shared with PCV7 and >96% for six additional serotypes. PMID- 19683658 TI - Pneumococcal pneumonia and influenza: a deadly combination. AB - Significant morbidity due to pneumococcal co-infection is associated with viral respiratory infections. Pneumonia is the leading cause of death in children worldwide. The incidence of clinical pneumonia among children in the United States decreased 39% following the introduction of a seven-valent pneumococcal conjugate vaccine (PCV). PCVs have also reduced hospitalisations associated with influenza in children. The majority of the mortality associated with the influenza pandemic of 1918 was attributable to bacterial infections, especially the pneumococcus. Vaccination with PCV for children and pneumococcal polysaccharide vaccine for adults should be considered essential to pandemic influenza preparedness. PMID- 19683660 TI - Recommendations for the management and follow-up of asthma exacerbations. Introduction. PMID- 19683661 TI - Noninvasive ventilation. PMID- 19683662 TI - Intubation and mechanical ventilation of the asthmatic patient in respiratory failure. PMID- 19683659 TI - Basic mechanisms and pathogenesis of venous thrombosis. AB - In 1856 Virchow proposed a triad of causes for venous thrombosis, postulating that stasis, changes in the vessel wall or changes in the blood could lead to thrombosis. We now know that abnormally high levels of some coagulation factors and defects in the natural anticoagulants contribute to thrombotic risk. Among these, factor V Leiden, which renders factor Va resistant to activated protein C, is the most prevalent with approximately 5% of the Caucasian population having this genetic alteration. These genetically controlled variants in coagulation factors work in concert with other risk factors, such as oral contraceptive use, to dramatically increase thrombotic risk. While these abnormalities in the blood coagulation proteins are associated with thrombotic disease propensity, they are less frequent contributors to thrombosis than age or cancer. Cancer increases thrombotic risk by producing tissue factor to initiate coagulation, by shedding procoagulant lipid microparticles or by impairing blood flow. Age is the strongest risk factor for thrombosis. Among possible reasons are fragility of the vessels potentially contributing to stasis, increased coagulation factor levels, impaired function of the venous valves, decreases in the efficacy of natural anticoagulants associated with the vessel wall, increased risk of immobilization and increased risk of severe infection. PMID- 19683663 TI - Anti-inflammatory treatment after discharge home from the emergency department in adults with acute asthma. PMID- 19683664 TI - Follow-up after acute asthma episodes: what improves future outcomes? PMID- 19683665 TI - Managing asthma exacerbations in the emergency department: summary of the National Asthma Education and Prevention Program Expert Panel Report 3 guidelines for the management of asthma exacerbations. PMID- 19683667 TI - Implication of snoRNA U50 in human breast cancer. AB - Deletion of chromosome 6q is frequent in breast cancer, and the deletion often involves a region in 6q14-q16. At present, however, the underlying tumor suppressor gene has not been established. Based on a recent study identifying snoRNA U50 as a candidate for the 6q14-16 tumor suppressor gene in prostate cancer, we investigated whether U50 is also involved in breast cancer. PCR-based approaches showed that U50 underwent frequent genomic deletion and transcriptional downregulation in cell lines derived from breast cancer. Mutation screening identified the same 2-bp deletion of U50 as in prostate cancer in both cell lines and primary tumors from breast cancer, and the deletion was both somatic and in germline. Genotyping of a cohort of breast cancer cases and controls for the mutation demonstrated that, while homozygous genotype of the mutation was rare, its heterozygous genotype occurred more frequently in women with breast cancer. Functionally, re-expression of U50 resulted in the inhibition of colony formation in breast cancer cell lines. These results suggest that noncoding snoRNA U50 plays a role in the development and/or progression of breast cancer. PMID- 19683669 TI - Effects of age on segregation of the X and Y chromosomes in cultured lymphocytes from Chinese men. AB - Chromosome malsegregation in binucleated lymphocytes is a useful endpoint to evaluate age effect on genetic stability. However, the investigations on chromosome malsegregation in binucleated lymphocytes from Chinese are scarce. In this study, peripheral blood lymphocytes were collected from 14 old (60-70 years) and 10 young (22-26 years) healthy Chinese men. To detect malsegregation of the sex chromosomes, multi-color fluorescence in situ hybridization (FISH) was performed on binucleated lymphocytes, cytokinesis-blocked by cytochalasin B at the first mitosis after phytohaemagglutinin stimulation. Compared with that in young men, a significant increase in frequencies of loss of chromosome X (9.2 +/- 3.2 per thousand vs. 1.1 +/- 0.9 per thousand, P < 0.001) and Y (2.5 +/- 1.9 per thousand vs. 0.2 +/- 0.3 per thousand, P < 0.001) was found in old men. Similarly, nondisjunction of chromosome X (16.5 +/- 3.4 per thousand vs. 3.5 +/- 1.1 per thousand, P < 0.001) and Y (7.2 +/- 2.6 per thousand vs. 2.4 +/- 1.3 per thousand, P < 0.001) occurred more frequently in old men than in young men. Regardless of donor's age, nondisjunction is more prevalent than loss for both chromosome X and Y. The frequencies of observed simultaneous malsegregation were relatively higher than the expected, suggesting an association between malsegregation. These results indicated that in Chinese men, malsegregation of the sex chromosomes increases with age in an associated fashion, and nondisjunction accounts for the majority of spontaneous chromosome malsegregation. PMID- 19683670 TI - Effects of splice sites on the intron retention in histamine H3 receptors from rats and mice. AB - In the alternative splicing, intron retention, of histamine H(3) receptors in rats and mice, the short transcript isoforms that are excised alternatively spliced introns are easily detected in a very low level in rats and are undetectable in mice using the regular PCR protocol. The retained introns have common 5' splice site and different 3' splice sites. The detailed mechanism for the special alternative splicing remains largely unclear. In this study, we developed a minigene splicing system to recapitulate natural alternative splicing of the receptors and investigated the effects of 5' and 3' splice sites on intron retention in HeLa cells. Mutating weak 5' and 3' splice sites of the alternatively spliced introns toward the canonical consensus sequences promoted the splicing of the corresponding introns in rat and mouse minigenes. The effect of splice site strength was context-dependent and much more significant for the 3' splice site of the longer alternative intron than for the 3' splice site of the shorter alternative intron and the common 5' splice sites; it was also more significant in the rat minigene than in the mouse minigene. Mutating the 3' splice site of the longer alternative intron resulted in almost complete splicing of the intron and made the corresponding isoform to become the nearly exclusive transcript in the rat minigene. PMID- 19683668 TI - Characterization of the promoter of phosphate transporter TaPHT1.2 differentially expressed in wheat varieties. AB - TaPHT1.2 is a functional, root predominantly expressed and low phosphate (Pi) inducible high-affinity Pi transporter in wheat, which is more abundant in the roots of P-efficient wheat genotypes (e.g., Xiaoyan 54) than in P-inefficient genotypes (e.g., Jing 411) under both Pi-deficient and Pi-sufficient conditions. To characterize TaPHT1.2 further, we genetically mapped a TaPHT1.2 transporter, TaPHT1.2-D1, on the long arm of chromosome 4D using a recombinant inbred line population derived from Xiaoyan 54 and Jing 411, and isolated a1,302 bp fragment of the TaPHT1.2-D1 promoter (PrTaPHT1.2-D1) from Xiaoyan 54. TaPHT1.2-D1 shows collinearity with OsPHT1.2 that has previously been reported to mediate the translocation of Pi from roots to shoots. PrTaPHT1.2-D contains a number of Pi starvation responsive elements, including P1BS, WRKY-binding W-box, and helix loop-helix-binding elements. PrTaPHT1.2-D1 was then used to drive expression of beta-glucuronidase (GUS) reporter gene in Arabidopsis through Agrobacterium mediated transformation. Histochemical analysis of transgenic Arabidopsis plants showed that the reporter gene was specifically induced by Pi-starvation and predominantly expressed in the roots. As there is only one SNP between the TaPHT1.2-D1 promoters of Xiaoyan 54 and Jing 411, and this SNP does not exist within the Pi-starvation responsive elements, the differential expression of TaPHT1.2 in Xiaoyan 54 and Jing 411 may not be caused by this SNP. PMID- 19683671 TI - vsx1 3' untranslated region-mediated translation difference at different developmental stages of goldfish embryos. AB - Visual system homeobox-1 (vsx1) is important in retinal progenitor proliferation, differentiation, and function maintenance of bipolar cells in vertebrates. Recent study in Xenopus laevis has shown that vsx1 3' untranslated region (3' UTR) can mediate cell-specific translation of vsx1 mRNA in the bipolar cells of the retinal inner nuclear layer (INL). vsx1 is also transcribed at the early developmental stages prior to eye formation and its spatiotemporal expression patterns are conserved in all the examined vertebrates. In order to determine whether the vsx1 3' UTR has a role in regulating the spatiotemporal expression of vsx1 during early embryogenesis, we constructed a vsx1 UTR-controlled green fluorescent protein (GFP) reporter gene system and examined the GFP expression pattern in goldfish, Carassius auratus, at different developmental stages. Our results indicated that both the vsx1 5' UTR and the vsx1 3' UTR remarkably repressed GFP expression at transcription level but did not regulate tissue specific translation at early developmental stages. GFP protein was ubiquitously expressed in the embryos injected with GFP-sensors containing vsx1 UTRs before 60 h post-fertilization (hpf). From hatching stage (72 hpf) onwards, however, GFP protein was specifically expressed in the bipolar cells of the retinal INL in the vsx1 3'UTR-GFP-sensor embryos, but was still ubiquitously expressed in the embryos injected with GFP-sensor lacking vsx1 3' UTR. These observations showed a significant difference of vsx1 3' UTR-mediated translation between early and late developmental stages and suggested that vsx1 3' UTR might not be involved in regulating the spatiotemporal expression of vsx1 until hatching stage during embryogenesis. PMID- 19683673 TI - The construction of a genetic linkage map of non-heading Chinese cabbage (Brassica campestris ssp. chinensis Makino). AB - Non-heading Chinese cabbage (Brassica campestris ssp. chinensis Makino) is one of the most important vegetables in eastern China. A genetic linkage map was constructed using 127 doubled haploid (DH) lines, and the DH population was derived from a commercial hybrid "Hanxiao" (lines SW-13 x L-118). Out of the 614 polymorphic markers, 43.49% were not assigned to any of the linkage groups(LGs). Chi-square tests showed that 42.67% markers were distorted from expected Mendelian segregation ratios, and the direction of distorted segregation was mainly toward the paternal parent L-118. After sequentially removing the markers that had an interval distance smaller than 1 cM from the upper marker, the overall quality of the linkage map was increased. Two hundred and sixty-eight molecular markers were mapped into 10 LGs, which were anchored to the corresponding chromosome of the B. rapa reference map based on common simple sequence repeat (SSR) markers. The map covers 973.38 cM of the genome and the average interval distance between markers was 3.63 cM. The number of markers on each LG ranged from 18 (R08) to 64 (R07), with an average interval distance within a single LG from 1.70 cM (R07) to 6.71 cM (R06). Among these mapped markers, 169 were sequence-related amplified polymorphism (SRAP) molecular markers, 50 were SSR markers and 49 were random amplification polymorphic DNA (RAPD) markers. With further saturation to the LG, the current map offers a genetic tool for loci analysis for important agronomic traits. PMID- 19683672 TI - Assessment of genetic diversity in broomcorn millet (Panicum miliaceum L.) using SSR markers. AB - The genetic diversity of 118 accessions of broomcorn millet (Panicum miliaceum L.), collected from various ecological areas, was analyzed. Using 46 SSR (Simple Sequence Repeat) polymorphic markers from rice, wheat, oat and barley, a total of 226 alleles were found, which exhibited moderate level of diversity. The number of alleles per primer ranged from two to nine, with an average of 4.91. The range of polymorphism information content (PIC) was 0.284-0.980 (average, 0.793). The expected heterozygosity (He) varied from 0.346 to 0.989, with an average of 0.834. The average coefficient of the genetic similarity of SSR markers among the 118 accessions was 0.609, and it ranged from 0.461 to 0.851. The UPGMA (Unweight Pair Group Method with Arithmetic Mean) clustering analysis at the genetic similarity value of 0.609 grouped the 118 accessions into five groups. Mantel test meant that geographical origin and genetic distance presented positive correlation. The clustering results were consistent with known information on ecological growing areas. The genetic similarity coefficient of the accessions in the Loess Plateau ecotype was significantly lower than those in the other ecotypes. It indicates that the highest level of genetic diversity occurred in the Loess Plateau, which is probably the original site of Panicum miliaceum. PMID- 19683674 TI - Fly antiviral RNA silencing and miRNA biogenesis claim ARS2. AB - In plants and invertebrates, small silencing RNAs function in antiviral defense and developmental patterning through pathways that were so far considered genetically distinct. In a recent issue of Cell, Sabin and colleagues report the identification of Drosophila Ars2, a protein required for both these small RNA mediated functions. PMID- 19683675 TI - Bacterial jailbreak sounds cellular alarm: phagosome membrane remnants trigger signaling. AB - Shigella flexneri and other invasive bacteria rupture the phagosome membrane and escape to the host cytoplasm. Now, Dupont et al. address the fate and signaling functions of pathogen vacuole remnants and show that these membrane fragments trigger host-cell-signaling responses, including polyubiquitination, autophagy, and pyroptosis. PMID- 19683676 TI - A gut feeling for microbes: getting it going between a parasite and its host. AB - We do not live in a sterile environment, but instead we are intimately associated with our commensal microbiota. Benson et al. (2009) suggest that host sensing of these microbes takes a pivotal role in getting the immune system up and running in response to a major intestinal parasite. PMID- 19683677 TI - Insect immunity: from pattern recognition to symbiont-mediated host defense. AB - The Jacques Monod conference "Insect Immunity in Action: From Fundamental Mechanisms of Host Defense to Resistance Against Infections in Nature," organized by Ulrich Theopold (Stockholm University, Sweden) and Dominique Ferrandon (CNRS, France), was held in May 2009 in Aussois, France. Here, we review key topics and concepts that were presented and highlight emerging trends in the field of insect immunity. PMID- 19683678 TI - Microbial telesensing: probing the environment for friends, foes, and food. AB - Bacterial-sensing circuits may be triggered by molecules originating from the environment (e.g., nutrients and chemoattractants). Bacteria also actively probe the environment for information by releasing molecular probes to measure conditions beyond the cell surface: a process known as telesensing. Perceiving the environment beyond is achieved by sensing environmentally induced changes in those probes, as occurs when a siderophore chelates an iron atom or a quorum sensing signal is inactivated by a specific enzyme or adsorbent. This information, captured by chemical and physical changes induced in specifically produced molecules transiting through the environment, enables bacteria to mount a contextually appropriate response. PMID- 19683679 TI - The S. Typhimurium effector SopE induces caspase-1 activation in stromal cells to initiate gut inflammation. AB - In the healthy intestinal mucosa, homeostasis between the immune system and commensal microflora prevents detrimental inflammatory responses. Infection with acute enteropathogens such as Salmonella enterica serovar Typhimurium disturbs this homeostasis and triggers inflammation, but the underlying mechanisms are poorly understood. We found that bacterial delivery or ectopic expression of the S. Typhimurium type III effector protein SopE, a known activator of host cellular Rho GTPases, led to proinflammatory caspase-1 activation and consequent maturation and secretion of the cytokine IL-1beta. In vivo, SopE triggered mucosal inflammation in wild-type but not caspase-1(-/-), IL-1R(-/-), or IL-18(-/ ) mice. Bone marrow chimeras indicated that caspase-1 was more important in stromal cells, most likely enterocytes, than in bone marrow-derived cells. SopE mediated caspase-1 activation in vitro was mediated by cellular Rho GTPases Rac-1 and Cdc42. These findings implicate SopE-driven Rho GTPase-mediated caspase-1 activation in stromal cells as a mechanism eliciting mucosal inflammation during S. Typhimurium infection. PMID- 19683680 TI - Shigella phagocytic vacuolar membrane remnants participate in the cellular response to pathogen invasion and are regulated by autophagy. AB - Intracellular pathogens like Shigella flexneri enter host cells by phagocytosis. Once inside, the pathogen breaks the vacuolar membrane for cytosolic access. The fate and function of the vacuolar membrane remnants are not clear. Examining Shigella-infected nonmyeloid cells, we observed that proteins associated with vacuolar membrane remnants are polyubiquinated, recruit the autophagy marker LC3 and adaptor p62, and are targeted to autophagic degradation. Further, inflammasome components and caspase-1 were localized to these membranes and correlated with dampened inflammatory response and necrotic cell death. In Atg4B mutant cells in which autophagosome maturation is blocked, polyubiquitinated proteins and P62 accumulated on membrane remnants, and as in autophagy-deficient Atg5(-/-) cells, the early inflammatory and cytokine response was exacerbated. Our results suggest that host membranes, after rupture by an invading cytoplasm targeted bacterium, contribute to the cellular responses to infection by acting as a signaling node, with autophagy playing a central role in regulating these responses. PMID- 19683681 TI - An essential role for RIG-I in toll-like receptor-stimulated phagocytosis. AB - Retinoic acid-inducible gene-I (RIG-I) plays an important role in antiviral response by recognizing double-stranded RNA. Here we demonstrate an unanticipated role of RIG-I in Toll-like receptor (TLR)-stimulated phagocytosis. Stimulation with lipopolysaccharide (LPS), a ligand of TLR4, induced the expression of RIG-I in macrophages. Depletion of RIG-I by RNAi or gene targeting inhibited the LPS induced phagocytosis of bacteria. Cellular processes involved in phagocytosis, such as small GTPase Cdc42/Rac1 activation, actin polymerization, and actin regulator Arp2/3 recruitment, were also impaired in RIG-I-deficient macrophages activated by LPS. Moreover, RIG-I(-/-) mice were found to be more susceptible to infection with Escherichia coli as compared to wild-type mice. Thus, the regulatory functions of RIG-I are strikingly broad, including a role not only in antiviral responses but in antibacterial responses as well. PMID- 19683682 TI - Reduced levels of protein tyrosine phosphatase CD45 protect mice from the lethal effects of Ebola virus infection. AB - Ebola virus (EBOV) infection of humans is a lethal but accidental dead-end event. Understanding resistance to EBOV in other species may help establish the basis of susceptibility differences among its hosts. Although rodents are resistant to EBOV, a murine-adapted variant is lethal when injected intraperitoneally into mice. We find that mice expressing reduced levels of the tyrosine phosphatase CD45 are protected against EBOV, whereas wild-type, CD45-deficient, or enzymatically inactive CD45-expressing mice succumbed to infection. Protection was dependent on CD8(+) T cells and interferon gamma. Reduced CD45-expressing mice retained greater control of gene expression and immune cell proliferation following EBOV infection, which contributed to reduced apoptosis, enhanced viral clearance, and increased protection against the virus. Together, these findings suggest that host susceptibility to EBOV is dependent on the delicate balance of immune homeostasis, which, as demonstrated here, can be determined by the levels of a single regulator. PMID- 19683683 TI - HIV-1 Nef interferes with host cell motility by deregulation of Cofilin. AB - HIV-1 Nef is a key factor in AIDS pathogenesis. Here, we report that Nef potently inhibits motility of fibroblasts and chemotaxis of HIV-1-infected primary human T lymphocytes toward the chemokines SDF-1alpha, CCL-19, and CCL-21 ex vivo. Furthermore, Nef inhibits guided motility of zebrafish primordial germ cells toward endogenous SDF-1a in vivo. These migration defects result from Nef mediated inhibition of the actin remodeling normally triggered by migratory stimuli. Nef strongly induces phosphorylation of cofilin, inactivating this evolutionarily conserved actin-depolymerizing factor that promotes cell motility when unphosphorylated. Nef-dependent cofilin deregulation requires association of Nef with the cellular kinase Pak2. Disruption of Nef-Pak2 association restores the cofilin phosphorylation levels and actin remodeling that facilitate cell motility. We conclude that HIV-1 Nef alters Pak2 function, which directly or indirectly inactivates cofilin, thereby restricting migration of infected T lymphocytes as part of a strategy to optimize immune evasion and HIV-1 replication. PMID- 19683684 TI - Gut commensal bacteria direct a protective immune response against Toxoplasma gondii. AB - Toxoplasma gondii is a universally distributed pathogen that infects over one billion people worldwide. Host resistance to this protozoan parasite depends on a Th1 immune response with potent production of the cytokines interleukin-12 and interferon gamma. Although Toll-like receptor 11 (TLR11) plays a major role in controlling Th1 immunity to this pathogen in mice, this innate immune receptor is nonfunctional in humans, and the mechanisms of TLR11-independent sensing of T. gondii remain elusive. Here, we show that oral infection by T. gondii triggers a TLR11-independent but MyD88-dependent Th1 response that is impaired in TLR2xTLR4 double knockout and TLR9 single knockout mice. These mucosal innate and adaptive immune responses to T. gondii rely on the indirect stimulation of dendritic cells by normal gut microflora. Thus, our results reveal that gut commensal bacteria can serve as molecular adjuvants during parasitic infection, providing indirect immunostimulation that protects against T. gondii in the absence of TLR11. PMID- 19683685 TI - Comparison of robotic and laparoscopic myomectomy. AB - OBJECTIVE: To compare surgical outcomes of patients with symptomatic leiomyomas after robot-assisted ("robotic") or laparoscopic myomectomy. STUDY DESIGN: Retrospective chart review of 81 patients undergoing robotic (n=40) or laparoscopic (n=41) myomectomy. Data included fibroid characteristics (number, weight, location, and pathologic findings), operating time, blood loss, complications, and postoperative hospitalization length. RESULTS: Patients undergoing laparoscopy had a significantly larger mean uterine size, larger mean size of the largest fibroid, and greater number of fibroids. When adjusted for uterine size and fibroid size and number, no significant differences were noted between robotic vs laparoscopic groups for mean operating time (141 vs 166 minutes), mean blood loss (100 vs 250 mL), intraoperative or postoperative complications (2% vs 20% and 11% vs 17%, respectively), hospital stay more than 2 days (12% vs 23%), readmissions, or symptom resolution. CONCLUSION: Short-term surgical outcomes were similar after robotic and laparoscopic myomectomy; long term outcomes were not assessed. PMID- 19683686 TI - Pelvic structure and function at 1 month compared to 7 months by dynamic magnetic resonance after vaginal birth. AB - OBJECTIVE: We sought to determine whether changes exist in location and movement of pelvic floor structures at 1 and 7 months postpartum. STUDY DESIGN: Midsagittal magnetic resonance images from 13 primiparous women with birth events associated with levator ani damage at early ( approximately 1 month) and late ( approximately 7 months) postpartum time points were analyzed. Pelvic floor structure locations at rest and displacements from rest to maximum Kegel and Valsalva were determined. Urogenital and levator hiatus diameters were measured as well. RESULTS: The perineal body was 7.1 mm and anal verge 7.9 mm higher at 7 months postpartum (P = .003). Both the urogenital and levator hiatus diameters were smaller at 7 months (P < .05). Displacement during Kegel and Valsalva was similar between the 2 time points. CONCLUSION: Resting locations of the perineal body and anal verge are higher at 7 months postpartum, but the amount of movement during Kegel or Valsalva does not change. PMID- 19683687 TI - Value of p16(INK4a) as a marker of progression/regression in cervical intraepithelial neoplasia grade 1. AB - OBJECTIVE: The objective of this study was to evaluate the usefulness of p16(INK4a) staining to classify cervical intraepithelial neoplasia grade 1 according to its progression/regression risk. STUDY DESIGN: Patients with a histologic diagnosis of cervical intraepithelial neoplasia grade 1 were prospectively recruited (n = 138). Simultaneous detection of high-risk human papillomaviruses and p16(INK4a) evaluation were performed. Follow-up was conducted every 6 months by cytology and colposcopy and annually by high-risk human papillomavirus testing, for at least 12 months (mean, 29.0). Progression was defined as a histologic diagnosis of cervical intraepithelial neoplasia grades 2-3, regression as a negative cytology and high-risk human papillomaviruses, and persistence as a cytologic result of low-grade squamous intraepithelial lesions and/or a positive test for high-risk human papillomaviruses. RESULTS: Progression was observed in 14 women (10.1%), 66 (47.6%) regressed, and 58 (42.0%) had a persistent disease. p16(INK4a) was positive in 77 (55.8%) initial biopsy specimens. Progression to cervical intraepithelial neoplasia grades 2-3 was identified in 14 of 77 (18.2%) women with positive and none of 61 (0.00%) women with negative p16(INK4a) immunostaining (P < .001). CONCLUSION: p16(INK4a) negative cervical intraepithelial neoplasia grade 1 lesions rarely progress and may benefit from a less intensive follow-up. PMID- 19683689 TI - Abdominal sacrocolpopexy and urinary incontinence: surgical planning based on urodynamics. AB - OBJECTIVE: The objective of the study was to evaluate the use of urodynamics to determine the need for incontinence surgery at the time of abdominal sacrocolpopexy (ASC). STUDY DESIGN: The records of 441 women undergoing ASC during 2005-2007 were reviewed. Group 1 consisted of 204 women (46.3%) with urodynamic stress incontinence (USI), including occult USI, who underwent incontinence surgery with ASC. Group 2 consisted of 237 women (53.7%) without USI who underwent ASC alone. Primary outcome measures were any complaint of postoperative incontinence (stress or urge) or new-onset urgency/frequency (UF). RESULTS: At a mean follow-up of 46.6 weeks, the overall rate of incontinence was low and similar for both groups (13.4% in group 1 and 13.3% in group 2 [P = .967]), as was new-onset UF: 18.6% in group 1 and 11.5% in group 2 (P = .195). CONCLUSION: Urodynamic evaluation appears to be useful in determining the need for incontinence surgery at the time of ASC. PMID- 19683690 TI - Ambulatory care related to female pelvic floor disorders in the United States, 1995-2006. AB - OBJECTIVE: The purpose of this study was to describe trends for pelvic floor disorder (PFD)-related ambulatory visits. STUDY DESIGN: Data were derived from the National Ambulatory Medical Care Survey and the National Hospital Ambulatory Care Survey. PFD-related visits were based on ICD-9 codes. We collapsed 12 survey years into 3 study periods (1995-1998, 1999-2002, 2003-2006) to evaluate numbers, rates, and trends for PFD-related visits. RESULTS: The average annual number of PFD-related visits was 3.9 million (95% confidence interval, 3.1-4.7). The annual rate of PFD-related visits per 1000 women was 35.2% in 1995-1998, 40.6% in 1999 2002, and 36.3% in 2003-2006. PFD visits represent 0.9% of all ambulatory visits for adult women in the United States. Women > or =60 years old had higher rates of PFD-related visits compared with women <60 years old. CONCLUSION: The annual number of PFD-related visits is significant and represents 0.9% of all ambulatory visits made by adult women in the United States. PMID- 19683691 TI - The evidence-based review of second-trimester abortion missed some important evidence. PMID- 19683693 TI - Pain associated with chorionic villus sampling: transabdominal vs transcervical approach. AB - OBJECTIVE: The purpose of this study was to compare anticipated and perceived pain that is associated with transabdominal and transcervical chorionic villus sampling (CVS). STUDY DESIGN: Women with singleton pregnancies who were undergoing CVS completed a preprocedure 0-10 visual analog scale (VAS; 0 = no pain, 10 = excruciating pain) for anticipated transabdominal and transcervical CVS-related pain. After the procedure, patients completed a VAS for perceived pain. RESULTS: One hundred twenty-one women underwent transabdominal (n = 98) or transcervical (n = 23) CVS. Anticipated pain was 4.5 +/- 2.0, which was similar in patients who ultimately underwent transabdominal (score, 4.6 +/- 3.8) or transcervical (score, 4.1 +/- 2.2) CVS. Postprocedure perceived pain was similar for transabdominal CVS in women with an abdominal wall thickness of <4 cm (score, 2.3 +/- 0.8) and transcervical CVS (score, 2.6 +/- 2.2) but was significantly greater for transabdominal CVS among women with an abdominal wall thickness of > or =4 cm (score, 5.6 +/- 1.2; P < .0001) and nulliparous women who had transcervical CVS (score, 4.3 +/- 2.1; P = .01). CONCLUSION: Transabdominal CVS is more painful in heavier women, and transcervical CVS is more painful in nulliparous women. PMID- 19683692 TI - Pregnancy: a "teachable moment" for weight control and obesity prevention. AB - Excessive gestational weight gain has been shown to relate to high-postpartum weight retention and the development of overweight and obesity later in life. Because many women are concerned about the health of their babies during pregnancy and are in frequent contact with their healthcare providers, pregnancy may be an especially powerful "teachable moment" for the promotion of healthy eating and physical activity behaviors among women. Initial research suggests that helping women gain the recommended amount during pregnancy through healthy eating and physical activity could make a major contribution to the prevention of postpartum weight retention. However, more randomized controlled trials with larger sample sizes are needed to identify the most effective and disseminable intervention. Providers have the potential to prevent high postpartum weight retention and future obesity by monitoring weight gain during pregnancy and giving appropriate advice about recommended amounts of gestational weight gain. PMID- 19683694 TI - Genetic association study of putative functional single nucleotide polymorphisms of genes in folate metabolism and spina bifida. AB - OBJECTIVE: We tested putative functional single nucleotide polymorphisms (SNPs) in genes that regulate the folate/homocysteine metabolism pathway for their contribution to spina bifida (SB) susceptibility. STUDY DESIGN: The study consisted of 610 unrelated simplex SB patient families. Genotypes of 46 SNPs located in the coding sequence or promoter region of 11 genes were investigated. Associations between transmission of alleles and SB in the offspring were examined using the reconstruction combined transmission disequilibrium test. RESULTS: Significant association of SNP rs5742905 in cystathionine-beta-synthase, rs1643649 in dihydrofolate reductase, rs2853533 in thymidylate synthetase, and rs3737965 in methylenetetrahydrofolate reductase was found (P = .015, .041, .021, and .007 respectively). CONCLUSION: Transmission disequilibrium of SNP alleles in cystathionine-beta-synthase, dihydrofolate reductase, methylenetetrahydrofolate reductase, and thymidylate synthetase confers an increased susceptibility to SB. PMID- 19683695 TI - Effect of estrogen replacement on the histologic response to polypropylene mesh implanted in the rabbit vagina model. AB - OBJECTIVE: The aim of this study was to determine the effect of estrogen replacement after ovariectomy on the histologic response to polypropylene mesh implanted in rabbit vagina. STUDY DESIGN: Thirty rabbits were assigned to 5 groups: sham laparotomy, ovariectomy, ovariectomy-preoperative estrogen, ovariectomy-postoperative estrogen, or ovariectomy-preoperative and postoperative estrogen. Rabbits underwent sham surgery or ovariectomy and were infused with vehicle or 17beta-estradiol (200 microg/d) for 4 weeks ("preoperative" estrogen). Polypropylene mesh was implanted in the posterior vaginal wall, and rabbits were infused with vehicle or "postoperative" estrogen for an additional 8 weeks. Grafts were harvested and underwent histologic evaluation. RESULTS: Vaginal atrophy in ovariectomized rabbits was reversed by estrogen replacement. Scores for inflammation (P = .33) and neovascularization (P = .23) at the graft site were not different among estrogen replacement groups, but estrogen administration was associated with increased collagen deposition (P = .005). CONCLUSION: Estrogen replacement administered for 8 weeks postoperatively increases collagen deposition into polypropylene mesh. PMID- 19683697 TI - Risk factors for the development of vesicovaginal fistula after incidental cystotomy at the time of a benign hysterectomy. AB - OBJECTIVE: We sought to evaluate risk factors for vesicovaginal fistula (VVF) after incidental cystotomy during benign hysterectomies. STUDY DESIGN: All benign hysterectomies between January 2000 and May 2004 were reviewed. Demographic and operative data were abstracted. Cystotomies were graded using the American Association for the Surgery of Trauma (AAST) system. Patients developing VVF after cystotomy were compared to those who did not. Categorical variables were analyzed with Fisher exact test while Student t test was used for continuous data. RESULTS: A total of 1317 benign hysterectomies were reviewed (46% abdominal, 48% vaginal, and 6% laparoscopically assisted vaginal). In all, 34 cystotomies occurred with 4 (11.7%) developing a VVF. Patients developing VVF were more likely to have an AAST grade V cystotomy (75% vs 7%; P = .004). Patients developing VVF trended toward greater tobacco use, larger uterine size, and more operative blood loss. CONCLUSION: Patients with an AAST grade V cystotomy are at increased risk for VVF formation. PMID- 19683696 TI - Evidence of increased oxidative stress and a change in the plasminogen activator inhibitor (PAI)-1 to PAI-2 ratio in early-onset but not late-onset preeclampsia. AB - OBJECTIVE: The aims of this study were to measure the degree of oxidative stress and alterations in plasminogen activator inhibitor (PAI) type 1 and PAI-2 ratio in women with early-onset and late-onset preeclampsia. STUDY DESIGN: A case control study was conducted in women with early-onset (24-32 weeks' gestation; n=18) and late-onset (35-42 weeks' gestation; n=20) preeclampsia and in control pregnant women at corresponding gestational weeks. Placenta, urine, and serum samples were collected. RESULTS: In early-onset preeclampsia, the median placental concentration of 8-iso-prostaglandin (PG)-F2alpha was higher and the PAI-1 to PAI-2 ratio higher than in early controls. These values did not differ between women with late-onset preeclampsia and their corresponding controls. Serum concentrations of 8-iso-PGF2alpha and vitamins C and E did not differ between cases and controls. CONCLUSION: Early-onset but not late-onset preeclampsia is associated with increased placental oxidative stress and increased PAI-1 to PAI-2 ratio. PMID- 19683698 TI - [Update of oral vitamin B12]. AB - OBJECTIVE: The objective of this review is to evaluate the usefulness of oral cobalamin (vitamin B12) treatment. MATERIAL AND METHOD: PubMed was systematically searched for English and French articles published from January 1990 to January 2007. RESULTS: Prospective randomized studies (n = 3), a systematic review by the Cochrane group (n = 1) and prospective studies in well-determined population (n = 5) provide evidence that oral cyanocobalamin therapy may adequately treat cobalamin deficiency in elderly patients. However, the current literature may not suggest a strategy in terms of the form (hydroxy- or cyanocobalamin), frequency and duration of the treatment. CONCLUSION: This present review confirms the previously reported efficacy of oral cyanocobalamin treatment in elderly patients. PMID- 19683699 TI - [Binge eating disorder and obesity: Epidemiological, clinical and psychopathological aspects. A study of an obese population in Sfax (Tunisia)]. AB - OBJECTIVES: To evaluate the prevalence of binge eating disorder (BED) in a population of obese patients, to appreciate the impact of obesity on BED through a comparison between the obese group and a control group, and to assess anxiety, depression and quality of life in obese patients with BED. PATIENTS AND METHODS: A cross-sectional study including 60 obese patients and 60 controls. BED was diagnosed using the Binge Eating Scale. Quality of life was assessed by the Quality Of Life, Obesity and Dietetics Scale, and depression and anxiety symptoms by the Hospital Anxiety and Depression Scale. RESULTS: The obese group had a higher prevalence of BED than the control group (40% versus 8.3%; p < 0.001; OR = 3.5). The average score of BES was also higher (p < 0.001). Obese patients with BED were younger (p = 0.034). BED was correlated with an early onset of obesity (p = 0.01; OR = 1.12), depression (p = 0.002), anxiety (p = 0.008) and a poorer quality of life. CONCLUSION: This study confirms the relationship between obesity and BED, which is correlated with a high prevalence of anxiety and depression and with a poorer quality of life. PMID- 19683700 TI - Air management and physiological responses during simulated firefighting tasks in a high-rise structure. AB - Air consumption, oxygen uptake (VO(2)), carbon dioxide output (VCO(2)) and respiratory exchange ratio (RER=VCO(2)/VO(2)) were measured directly from the self-contained breathing apparatus (SCBA) as 36 professional firefighters (three women) completed scenarios of high-rise stair climbing and fifth floor search and rescue. During stair climbing VO(2) was 75+/-8% VO(2max) (mean+/-SD), RER=1.10+/ 0.10, and heart rate=91+/-3% maximum (based on maximum treadmill data). Firefighters stopped climbing on consuming 55% of the air cylinder then descended. In the fifth floor search and rescue VO(2) was slightly lower than stair climbing but RER remained elevated (1.13+/-0.12) reflecting high anaerobic metabolism. The first low air alarm sounded, indicating 25% of the air remaining in a "30-min cylinder", during the stair climb at 8 min with 19 of 36 sounding before 12 min. Aggressive air management strategies are required for safety in high-rise firefighting. PMID- 19683701 TI - A portable device for the assessment of dynamic visual acuity. AB - Dynamic visual acuity (DVA) thresholds are among the few visual functions predictive of automobile crashes. DVA is also sensitive to alcohol and aging. However, measuring DVA is awkward because there is no standardized, efficient, flexible apparatus for DVA assessment. In this project, we developed a prototype of an automated, portable DVA system using a low-energy laser, and we compared this laser DVA with the traditional device in two within-subjects, repeated measures designs. The two studies included 48 participants (22 males and 26 females with an average age of 18.33 years). The most important findings were that: (1) retest reliabilities of the two DVA devices were comparable and higher with the laser; (2) average correlations between the two devices were r=0.62 (p<0.01) and r=0.65 (p<0.01) for the two designs respectively; and (3) after correction for reliability attenuation these improved to r=0.92 and r=0.78. These findings indicate that a flexible DVA laser device can be developed to measure the same construct as the more traditional bulky DVA device. PMID- 19683702 TI - Recombinant adeno-associated virus serotype 2 (rAAV2)-An efficient vector for gene delivery in condylar cartilage, glenoid fossa and TMJ disc in an experimental study in vivo. AB - OBJECTIVE: To elaborate whether rAAV2 can be used for future TMJ gene therapy, we examined the infection efficiencies of rAAV2 in vitro, and the transgene expression pattern mediated by rAAV2 in glenoid fossa, TMJ disc and condylar cartilage in vivo. MATERIALS AND METHODS: Different dosages of rAAV2-eGFP (MOI: 5 x 10(4), 1 x 10(4), 5 x 10(3)) were applied to primary cultured condylar chondrocytes of rats. Infection efficiencies were analysed by FACSCalitur at different time points. Vastatin, a molecule not naturally expressed in TMJ, was used as a reporter for detection of rAAV2 mediated transgene expression in vivo. Thirty SD rats were injected with either rAAV2-sec-Vastatin (experimental group) or rAAV2-eGFP (control group) into both sides of TMJ. They were sacrificed at the indicated time (7, 14, 21, 30 and 60 days of injection) and the TMJ samples were collected for RT-PCR and immunostaining analysis. RESULTS: High dosage (MOI 5 x 10(4)) of rAAV2-eGFP can achieve desirable transduction efficiencies in vitro after 5 days. Transgene expression of rAAV-sec-Vastatin persisted for about 21 days in glenoid fossa, around 7 days in TMJ disc and at least 60 days in condylar cartilage in vivo. In condylar cartilage, transgene expression was found in the proliferative layer and chondroblast layer (day 7), chondrocyte layer (day 14), pre-hypertrophic and hypertrophic layer (day 21), hypertrophic layer and deep hypertrophic layer (day 30 and 60). CONCLUSION: Recombinant AAV2 could be considered as a promising vector for gene therapy in TMJ which can mediate therapeutic gene expression in glenoid fossa, articular disc and condylar cartilage in vivo. PMID- 19683703 TI - School intervention for promoting psychological well-being in adolescence. AB - OBJECTIVE: to test the efficacy of a new school program for the promotion of psychological well-being. In this study a school program for promoting psychological well-being has been compared to an attention-placebo intervention in a high school setting. METHODS: Nine classes (227 students) were randomly assigned to: a) Well-Being intervention (5 classes); b)attention-placebo (4 classes). Assessment was performed at pre and post-intervention, and after six months using: 1) Symptom Questionnaire (SQ); 2) Psychological Well-Being Scales (PWB); 3) Revised Children's Manifest Anxiety Scale (RCMAS). RESULTS: A significant effect of WB school intervention in improving Personal Growth (PWB), and in decreasing distress (Somatization (SQ), Physical Well-being (SQ), Anxiety (SQ), and RCMAS Physiological Anxiety) emerged. CONCLUSIONS: A school intervention based on promoting positive emotions and well-being was effective not only in increasing psychological well-being among adolescents, but also in decreasing distress, in particular anxiety and somatization. PMID- 19683705 TI - Etanercept for sleep in patients with alcohol use disorder--mechanisms need to be elucidated. PMID- 19683706 TI - Increased volume of the striatum in psychopathic individuals. AB - BACKGROUND: The corpus striatum, comprised of the caudate, putamen, and globus pallidus, plays an important role in reward processing and may be involved in the pathophysiology of antisocial behavior. Few studies have explored whether differences are present in the striatum of antisocial individuals. Here, we examine the structure of the striatum in relation to psychopathy. METHODS: Using a case-control design, we examined the volume of the striatum in psychopathic individuals compared with control subjects matched for age, sex, ethnicity, and substance dependence. Twenty-two psychopathic individuals assessed using the Psychopathy Checklist-Revised and 22 comparison subjects underwent structural magnetic resonance imaging. Volumes of the left and right lenticular nucleus (putamen and globus pallidus), caudate head, and caudate body were assessed and the psychopathic and control groups were compared. RESULTS: Psychopathic individuals showed a 9.6% increase in striatum volumes. Analyses of subfactors of psychopathy revealed that caudate body volumes were primarily associated with the interpersonal and affective features of psychopathy, while caudate head volumes were primarily associated with the impulsive, stimulation-seeking features. CONCLUSIONS: These findings provide new evidence for differences in the striatum of psychopathic individuals. This structural difference may partially underlie the reward-seeking and decision-making deficits associated with psychopathy. PMID- 19683708 TI - Synthesis and X-ray structure of a C5-C4-linked glucofuranose-oxazolidin-2-one. AB - The formation of (4R)-4-carbamoyl-4-[(4R)-3-O-benzyl-1,2-O-isopropylidene-beta-l threofuranos-4-C-yl]-oxazolidin-2-one instead of expected imidazolidin-2,4-dione (hydantoin) derivative from 5-amino-5-cyano-5-deoxy-3-O-benzyl-1,2-O isopropylidene-alpha-d-glucofuranose or 3-O-benzyl-1,2-O-isopropylidene-alpha-d xylo-hexofuranos-5-ulose under Bucherer-Bergs reaction conditions is reported. Single crystal X-ray diffraction data revealed that (3)T(4) is the prefered conformation for the furanose ring, while E(2) and (2)T(1) conformations are adopted by the 1,3-dioxolane and 2-oxazolidinone five-membered rings, respectively. PMID- 19683707 TI - An innovative method for oxidative degradation of chitosan with molecular oxygen catalyzed by metal phthalocyanine in neutral ionic liquid. AB - A novel aqueous solution-ionic liquid biphasic catalytic system was established for the oxidative degradation of chitosan under mild conditions. In this process, the environmentally acceptable and inexpensive molecular oxygen was first used as oxidant, the metal phthalocyanine was immobilized in ionic liquid as catalyst, and the aqueous solution as medium carried the reactants and the products. Under vigorous stirring and heating, the reactants fully contacted the catalysts in the emulsion and chitosan efficiently degraded into water-soluble materials. At the end of the reaction, the catalytic system could be easily separated by simple decantation and could also be reused in subsequent runs without apparent change in activity. These characters are in favor of the elimination of pollution and the reduction of the economic cost in the large-scale production of the water soluble chitosan derivatives in chemical industry. PMID- 19683709 TI - Adaptive compression algorithm from projections: application on medical greyscale images. AB - Image compression plays a crucial role in medical imaging, allowing efficient manipulation, storage, and transmission of binary, grey-scale, or colour images. Nevertheless, in medical applications the need to conserve the diagnostic validity of the image requires the use of lossless compression methods, producing low compression factors. In this paper, a novel near-lossless compression algorithm from projections, which almost eliminates both redundant information and noise from a greyscale image while retaining all relevant structures and producing high compression factors, is proposed. The algorithm is tested on experimental medical greyscale images from different modalities and different body districts and results are reported. PMID- 19683710 TI - Acquired prosopagnosia abolishes the face inversion effect. AB - Individual faces are notoriously difficult to recognize when they are presented upside-down. Since acquired prosopagnosia (AP) has been associated with an impairment of expert face processes, a reduced or abolished face inversion effect (FIE) is expected in AP. However, previous studies have incongruently reported apparent normal effects of inversion, a decreased or abolished FIE, but also a surprisingly better performance for inverted faces for some patients. While these discrepant observations may be due to the variability of high-level processes impaired, a careful look at the literature rather suggests that the pattern of FIE in prosopagnosia has been obscured by a selection of patients with associated low-level defects and general visual recognition impairments, as well as trade offs between accuracy and correct RT measures. Here we conducted an extensive investigation of upright and inverted face processing in a well-characterized case of face-selective AP, PS (Rossion et al., 2003). In 4 individual face discrimination experiments, PS did not present any inversion effect at all, taking into account all dependent measures of performance. However, she showed a small inversion cost for individualizing members of a category of non-face objects (cars), just like normal observers. A fifth experiment with personally familiar faces to recognize confirmed the lack of inversion effect for PS. Following the present report and a survey of the literature, we conclude that the FIE is generally absent, or at least clearly reduced following AP. We also suggest that the paradoxical superior performance for inverted faces observed in rare cases may be due to additional upper visual field defects rather than to high-level competing visual processes. These observations are entirely compatible with the view that AP is associated with a disruption of a process that is also abolished following inversion: the holistic representation of individual exemplars of the face class. PMID- 19683711 TI - A novel surgical treatment for cesarean scar pregnancy: laparoscopically assisted operative hysteroscopy. AB - OBJECTIVE: To describe laparoscopically assisted hysteroscopy as a unique surgical intervention for a cesarean section scar ectopic pregnancy. DESIGN: Case report. SETTING: University hospital. PATIENT(S): A 44-year-old woman, pregnant at 5 weeks and 6 days' gestational age with a cesarean section scar ectopic pregnancy. INTERVENTION(S): The patient underwent serial transvaginal ultrasound examinations with Doppler flow studies, followed by a laparoscopically assisted operative hysteroscopy for evacuating the cesarean scar ectopic pregnancy. MAIN OUTCOME MEASURE(S): Conservation of the uterus, fertility preservation. RESULT(S): Successful conservative surgical treatment of cesarean section scar ectopic pregnancy. CONCLUSION(S): Conservative laparoscopically assisted operative hysteroscopy can be used successfully in hemodynamically stable patients with a cesarean section scar ectopic pregnancy. PMID- 19683713 TI - Magnifying the endoscopic appearance of Cronkhite-Canada syndrome. PMID- 19683712 TI - Mediastinal-esophageal fistulae after EUS-FNA of tuberculosis of the mediastinum. PMID- 19683714 TI - Rectal fistulae resulting from treatment with sorafenib and bevacizumab. PMID- 19683716 TI - Surface properties and aggregate morphology of partially fluorinated carboxylate type anionic gemini surfactants. AB - Three anionic homologues of a novel partially fluorinated carboxylate-type anionic gemini surfactant, N,N'-di(3-perfluoroalkyl-2-hydroxypropyl)-N,N' diacetic acid ethylenediamine (2C(n)(F) edda, where n represents the number of carbon atoms in the fluorocarbon chain (4, 6, and 8)) were synthesized. In these present gemini surfactants, the relatively small carboxylic acid moieties form hydrophilic head groups. The surface properties or structures of the aggregates of these surfactants are strongly influenced by the nonflexible fluorocarbons and small head groups; this is because these surfactants have a closely packed molecular structure. The equilibrium surface tension properties of these surfactants were measured at 298.2K for various fluorocarbon chain lengths. The plot of the logarithm of the critical micelle concentration (cmc) against the fluorocarbon chain lengths for 2C(n)(F) edda (n=4, 6, and 8) showed a minimum for n=6. Furthermore, the lowest surface tension of 2C(6)(F) edda at the cmc was 16.4mNm(-1). Such unique behavior has not been observed even in the other fluorinated surfactants. Changes in the shapes and sizes of these surfactant aggregate with concentration were investigated by dynamic light scattering and transmission electron microscopy (TEM). The TEM micrographs showed that in an aqueous alkali solution, 2C(n)(F) edda mainly formed aggregates with stringlike (n=4), cagelike (n=6), and distorted bilayer structures (n=8). The morphological changes in the aggregates were affected by the molecular structure composed of nonflexible fluorocarbon chains and flexible hydrocarbon chains. PMID- 19683715 TI - Quality of faculty, students, curriculum and resources for nursing doctoral education in Korea: a focus group study. AB - BACKGROUND: The rapidly increasing number of nursing doctoral programs has caused concern about the quality of nursing doctoral education, including in Korea. OBJECTIVES: To describe the perceived quality of Korean nursing doctoral education in faculty, student, curriculum and resources. DESIGN: Focus group. SETTINGS: Fourteen Korean nursing doctoral programs that are research focused and include coursework. PARTICIPANTS: Four groups of deans, faculty, students and graduates; students completed three semesters of doctoral program; and graduates completed doctoral programs within the most recent 3 years. METHODS: Focus groups examined the strengths and weaknesses of faculty, students, curriculum, and resources. RESULTS: Faculty strengths were universities' recognition of faculty research/scholarship and the ability of faculty to attract extramural funding. Faculty weaknesses were aging faculty; high faculty workload; insufficient number of faculty; and teaching without expertise in nursing theories. Student strengths were diverse student backgrounds; multidisciplinary dissertation committee members, and opportunities to socialize with peers and graduates/faculty. Students' weaknesses were overproduction of PhDs with low academic quality; a lower number and quality of doctoral applicants; and lack of full-time students. Curriculum strengths were focusing on specific research areas; emphasis on research ethics; and multidisciplinary courses. Curriculum weaknesses were insufficient time for curriculum development; inadequate courses for core research competencies; and a lack of linkage between theory and practice. Resources strengths were inter-institutional courses with credit transfer. Weaknesses were diminished university financial support for graduate students and limited access to school facilities. Variations in participant groups (providers [deans and faculty] vs. receivers [students and graduates]) and geographical location (capital city vs. regional) were noted on all the four components. CONCLUSIONS: The quality characteristics of faculty, students, curriculum, and resources identified in this first systematic evaluation of the quality of nursing doctoral education can inform nursing schools, universities, and policy makers about areas for improvement in Korea and possibly in the world. Geographical variations found in these four components of doctoral education warrant attention by policy-makers in Korea. PMID- 19683717 TI - A modified Cassie-Baxter relationship to explain contact angle hysteresis and anisotropy on non-wetting textured surfaces. AB - The Cassie-Baxter model is widely used to predict the apparent contact angles obtained on composite (solid-liquid-air) superhydrophobic interfaces. However, the validity of this model has been repeatedly challenged by various research groups because of its inherent inability to predict contact angle hysteresis. In our recent work, we have developed robust omniphobic surfaces which repel a wide range of liquids. An interesting corollary of constructing such surfaces is that it becomes possible to directly image the solid-liquid-air triple-phase contact line on a composite interface, using an electron microscope with non-volatile organic liquids or curable polymers. Here, we fabricate a range of model superoleophobic surfaces with controlled surface topography in order to correlate the details of the local texture with the experimentally observed apparent contact angles. Based on these experiments, in conjunction with numerical simulations, we modify the classical Cassie-Baxter relation to include a local differential texture parameter which enables us to quantitatively predict the apparent advancing and receding contact angles, as well as contact angle hysteresis. This quantitative prediction also allows us to provide an a priori estimation of roll-off angles for a given textured substrate. Using this understanding we design model substrates that display extremely small or extremely large roll-off angles, as well as surfaces that demonstrate direction dependent wettability, through a systematic control of surface topography and connectivity. PMID- 19683718 TI - Freeze-thaw stability of water-in-oil emulsions. AB - Factors influencing water-in-oil emulsion stability during freeze/thaw-cycling, namely interfacial crystallization vs. network crystallization and the sequence of crystallization events (i.e., dispersed vs. continuous phase or vice versa), are assessed. We show that destabilization is most apparent with a liquid-state emulsifier and a continuous oil phase that solidifies prior to the dispersed phase. Emulsions stable to F/T-cycling are obtained when the emulsifier crystallizes at the oil-water interface or in emulsions where the continuous phase crystallizes after the dispersed aqueous phase. The materials used are two food-grade oil-soluble emulsifiers - polyglycerol polyricinoleate (PGPR) and glycerol monostearin (GMS) and two continuous oil phases with differing crystallization temperatures - canola oil and coconut oil. Emulsion stability is assessed with pulsed field gradient NMR droplet size analysis, sedimentation, microscopy and differential scanning calorimetry. This study demonstrates the sequence of crystallization events and the physical state of the surfactant at the oil-water interface strongly impact the freeze-thaw stability of water-in-oil emulsions. PMID- 19683719 TI - Ternary [Al(2)O(3)-electrolyte-Cu(2+)] species: EPR spectroscopy and surface complexation modeling. AB - Cu(2+) binding on gamma-Al(2)O(3) is modulated by common electrolyte ions such as Mg(2+), SO(4)(2-), and PO(4)(3-) in a complex manner: (a) At high concentrations of electrolyte ions, Cu(2+) uptake by gamma-Al(2)O(3) is inhibited. This is partially due to bulk ionic strength effects and, mostly, due to direct competition between Mg(2+) and Cu(2+) ions for the SO(-) surface sites of gamma Al(2)O(3). (b) At low concentrations of electrolyte ions, Cu(2+) uptake by gamma Al(2)O(3) can be enhanced. This is due to synergistic coadsorption of Cu(2+) and electrolyte anions, SO(4)(2-) and PO(4)(3-)(.) This results in the formation of ternary surface species (SOH(2)SO(4)Cu)(+), (SOH(2)PO(4)Cu), and (SOH(2)HPO(4)Cu)(+) which enhance Cu(2+) uptake at pH<6. The effect of phosphate ions may be particularly strong resulting in a 100% Cu uptake by the oxide surface. (c) EPR spectroscopy shows that at pH<>pH(PZC) Cu(2+) may coordinate to two SO(-) groups. At pH<